The Scientific World Journal: Biotechnology The latest articles from Hindawi Publishing Corporation © 2016 , Hindawi Publishing Corporation . All rights reserved. In Vitro Seeds Germination and Seedling Growth of Bambara Groundnut (Vigna subterranea (L.) Verdc. (Fabaceae)) Tue, 13 Oct 2015 12:06:07 +0000 Bambara groundnut (Vigna subterranea (L.) Verdc.) is an indigenous grain legume. It occupies a prominent place in the strategies to ensure food security in sub-Saharan Africa. Development of an efficient in vitro regeneration system, a prerequisite for genetic transformation application, requires the establishment of optimal conditions for seeds germination and plantlets development. Three types of seeds were inoculated on different basal media devoid of growth regulators. Various strengths of the medium of choice and the type and concentration of carbon source were also investigated. Responses to germination varied with the type of seed. Embryonic axis (EA) followed by seeds without coat (SWtC) germinated rapidly and expressed a high rate of germination. The growth performances of plantlets varied with the basal medium composition and the seeds type. The optimal growth performances of plants were displayed on half strength MS basal medium with SWtC and EA as source of seeds. Addition of 3% sucrose in the culture medium was more suitable for a maximum growth of plantlets derived from EA. Mongomaké Koné, Tchoa Koné, Nakpalo Silué, André Brahima Soumahoro, and Tanoh Hilaire Kouakou Copyright © 2015 Mongomaké Koné et al. All rights reserved. Variations in Hormones and Antioxidant Status in Relation to Flowering in Early, Mid, and Late Varieties of Date Palm (Phoenix dactylifera) of United Arab Emirates Tue, 16 Jun 2015 13:44:10 +0000 The present study was carried out to assess the status of various hormones responsible for the flower induction of Nagal, Lulu, and Khalas date palm varieties in UAE. The nonenzymatic antioxidant compounds and the antioxidant enzymatic activities at preflowering, flowering, and postflowering stages of the date palm varieties were quantified. The ABA and zeatin concentrations were found to be significantly higher during the preflowering stage but gradually decreased during the flowering period and then increased after the flowering stage. Gibberellic acid (GA) concentrations were significantly higher in the early flowering varieties and higher levels of ABA may contribute to the delayed flowering in mid and late varieties. The results on hormone profiling displayed a significant variation between seasons (preflowering, flowering, and postflowering) and also between the three date palms (early, mid, and late flowering varieties). Ascorbic acid (AA) concentration was low at the preflowering stage in the early flowering Nagal (0.694 mg/g dw), which is similar with the late flowering Lulu variety (0.862 mg/g dw). However, Khalas variety showed significantly higher amount of AA content (7.494 mg/g dw) at the preflowering stage when compared to other varieties. In flowering stage, Nagal (0.814 mg/g dw) and Lulu (0.963 mg/g dw) were similar with respect to the production of AA, while the mid flowering variety showed significantly higher amount of AA (9.358 mg/g dw). The Khalas variety produced the highest tocopherol at 4.78 mg/g dw compared to Nagal and Lulu, at 1.997 and 1.908 mg/g dw, respectively, during the preflowering stage. In Nagal variety, the content of reduced glutathione (GSH) at the preflowering stage was 0.507 mg/g dw, which was not significantly different from the flowering and postflowering stages at 0.4 and 0.45 mg/g dw, respectively. The GSH was significantly higher in Khalas compared to Nagal and Lulu varieties, at 1.321 mg/g dw in the preflowering phase followed by 3.347 mg/g dw and 2.349 mg/g dw at the flowering and postflowering phases, respectively. Catalase activity increased with different stages of growth. The lowest catalase activity was observed at the preflowering stage in Khalas (0.116), with similar observations noted during flowering (0.110) and postflowering stage. This study provides an insight into the possible roles of endogenous hormones and antioxidants and in the activities of antioxidant enzymes in the regulation of flower development in date palm varieties. Abdul J. Cheruth, Shyam S. Kurup, and Sreeramanan Subramaniam Copyright © 2015 Abdul J. Cheruth et al. All rights reserved. Structural, Morphological, and Electron Transport Studies of Annealing Dependent In2O3 Dye-Sensitized Solar Cell Thu, 04 Jun 2015 09:04:57 +0000 Indium oxide (In2O3) thin films annealed at various annealing temperatures were prepared by using spin-coating method for dye-sensitized solar cells (DSSCs). The objective of this research is to enhance the photovoltaic conversion efficiency in In2O3 thin films by finding the optimum annealing temperature and also to study the reason for high and low performance in the annealed In2O3 thin films. The structural and morphological characteristics of In2O3 thin films were studied via XRD patterns, atomic force microscopy (AFM), field-emission scanning electron microscopy (FESEM), EDX sampling, and transmission electron microscopy (TEM). The annealing treatment modified the nanostructures of the In2O3 thin films viewed through FESEM images. The In2O3-450°C-based DSSC exhibited better photovoltaic performance than the other annealed thin films of 1.54%. The electron properties were studied by electrochemical impedance spectroscopy (EIS) unit. The In2O3-450°C thin films provide larger diffusion rate, low recombination effect, and longer electron lifetime, thus enhancing the performance of DSSC. S. Mahalingam, H. Abdullah, S. Shaari, A. Muchtar, and I. Asshari Copyright © 2015 S. Mahalingam et al. All rights reserved. Wing Morphometry and Acoustic Signals in Sterile and Wild Males: Implications for Mating Success in Ceratitis capitata Thu, 14 May 2015 13:49:03 +0000 The sterile insect technique (SIT) is widely utilized in the biological control of fruit flies of the family Tephritidae, particularly against the Mediterranean fruit fly. This study investigated the interaction between mating success and morphometric variation in the wings and the production of acoustic signals among three male groups of Ceratitis capitata (Wiedemann): (1) wild males, (2) irradiated with Co-60 (steriles), and (3) irradiated (steriles) and treated with ginger oil. The canonical variate analysis discriminated two groups (males irradiated and males wild), based on the morphological shape of the wings. Among males that emit buzz signals, wild males obtained copulation more frequently than males in Groups 2 and 3. The individuals of Group 3 achieved more matings than those in Group 2. Wild males displayed lower pulse duration, higher intervals between pulses, and higher dominant frequency. Regarding the reproductive success, the morphological differences in the wings’ shape between accepted and nonaccepted males are higher in wild males than in the irradiated ones. The present results can be useful in programs using the sterile insect technique for biological control of C. capitata. João Maria Gomes Alencar de Souza, Paulo Augusto de Lima-Filho, Wagner Franco Molina, Lúcia Maria de Almeida, Milson Bezerra de Gouveia, Francisco Pepino de Macêdo, Raul Alberto Laumann, and Beatriz Aguiar Jordão Paranhos Copyright © 2015 João Maria Gomes Alencar de Souza et al. All rights reserved. Effects of Droplet-Vitrification Cryopreservation Based on Physiological and Antioxidant Enzyme Activities of Brassidium Shooting Star Orchid Wed, 11 Mar 2015 10:23:11 +0000 Protocorm-like bodies (PLBs) of Brassidium Shooting Star orchid were successfully cryopreserved using droplet-vitrification method. Vitrification based cryopreservation protocol is comprised of preculture, osmoprotection, cryoprotection, cooling, rewarming, and growth recovery and each and every step contributes to the achievement of successful cryopreservation. In order to reveal the lethal and nonlethal damage produced by cryopreservation, histological observation, scanning electron microscopy (SEM), and biochemical analysis were carried out in both cryopreserved and noncryopreserved PLBs of Brassidium Shooting Star orchid comparing with the control PLBs stock culture. Histological and scanning electron microscopy analyses displayed structural changes in cryopreserved PLBs due to the impact of cryoinjury during exposure to liquid nitrogen. Total soluble protein significantly increased throughout the dehydration process and the highest value was achieved when PLBs were stored in liquid nitrogen. Ascorbate peroxidase (APX) and catalase (CAT) showed the highest enzyme activities in both dehydration and cryostorage treatments indicating that stress level of PLBs was high during these stages. Safrina Rahmah, Safiah Ahmad Mubbarakh, Khor Soo Ping, and Sreeramanan Subramaniam Copyright © 2015 Safrina Rahmah et al. All rights reserved. Resistance to and Accumulation of Heavy Metals by Actinobacteria Isolated from Abandoned Mining Areas Wed, 11 Feb 2015 11:47:11 +0000 Accumulation of high concentrations of heavy metals in environments can cause many human health risks and serious ecological problems. Nowadays, bioremediation using microorganisms is receiving much attention due to their good performance. The aim of this work is to investigate heavy metals resistance and bioaccumulation potential of actinobacteria strains isolated from some abandoned mining areas. Analysis of mining residues revealed that high concentration of zinc “Zn” was recorded in Sidi Bouatman, Arbar, and Bir Nhass mining residues. The highest concentration of lead “Pb” was found in Sidi Bouatman. Copper “Cu,” cadmium “Cd,” and chromium “Cr” were found with moderate and low concentrations. The resistance of 59 isolated actinobacteria to the five heavy metals was also determined. Using molecular identification 16S rRNA, these 27 isolates were found to belong to Streptomyces and Amycolatopsis genera. The results showed different levels of heavy metal resistance; the minimum inhibitory concentration (MIC) recorded was 0.55 for Pb, 0.15 for Cr, and 0.10 mg·mL−1 for both Zn and Cu. Chemical precipitation assay of heavy metals using hydrogen sulfide technic (H2S) revealed that only 27 isolates have a strong ability to accumulate Pb (up to 600 mg of Pb per g of biomass for Streptomyces sp. BN3). Soraia El Baz, Mohamed Baz, Mustapha Barakate, Lahcen Hassani, Abdelhay El Gharmali, and Boujamâa Imziln Copyright © 2015 Soraia El Baz et al. All rights reserved. Decolorization of Anthraquinonic Dyes from Textile Effluent Using Horseradish Peroxidase: Optimization and Kinetic Study Mon, 19 Jan 2015 07:30:06 +0000 Two anthraquinonic dyes, C.I. Acid Blue 225 and C.I. Acid Violet 109, were used as models to explore the feasibility of using the horseradish peroxidase enzyme (HRP) in the practical decolorization of anthraquinonic dyes in wastewater. The influence of process parameters such as enzyme concentration, hydrogen peroxide concentration, temperature, dye concentration, and pH was examined. The pH and temperature activity profiles were similar for decolorization of both dyes. Under the optimal conditions, 94.7% of C.I. Acid Violet 109 from aqueous solution was decolorized (treatment time 15 min, enzyme concentration 0.15 IU/mL, hydrogen peroxide concentration 0.4 mM, dye concentration 30 mg/L, pH 4, and temperature 24°C) and 89.36% of C.I. Acid Blue 225 (32 min, enzyme concentration 0.15 IU/mL, hydrogen peroxide concentration 0.04 mM, dye concentration 30 mg/L, pH 5, and temperature 24°C). The mechanism of both reactions has been proven to follow the two substrate ping-pong mechanism with substrate inhibition, revealing the formation of a nonproductive or dead-end complex between dye and HRP or between H2O2 and the oxidized form of the enzyme. Both chemical oxygen demand and total organic carbon values showed that there was a reduction in toxicity after the enzymatic treatment. This study verifies the viability of use of horseradish peroxidase for the wastewaters treatment of similar anthraquinonic dyes. Nataša Ž. Šekuljica, Nevena Ž. Prlainović, Andrea B. Stefanović, Milena G. Žuža, Dragana Z. Čičkarić, Dušan Ž. Mijin, and Zorica D. Knežević-Jugović Copyright © 2015 Nataša Ž. Šekuljica et al. All rights reserved. Impact Assessment of Mercury Accumulation and Biochemical and Molecular Response of Mentha arvensis: A Potential Hyperaccumulator Plant Sun, 11 Jan 2015 09:52:25 +0000 The present study was focused on examining the effect of Hg oxidative stress induced physiochemical and genetic changes in M. arvensis seedlings. The growth rate of Hg treated seedlings was decreased to 56.1% and 41.5% in roots and shoots, respectively, compared to the control. Accumulation of Hg level in both roots and shoots was increased with increasing the concentration of Hg. Superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) activities were found to be increased with increasing the Hg concentration up to 20 mg/L; however, it was decreased at 25 mg/L Hg concentration. The POX enzyme activity was positively correlated with Hg dose. The changes occurring in the random amplification of ploymorphic DNA (RAPD) profiles generated from Hg treated seedlings included variations in band intensity, disappearance of bands, and appearance of new bands compared with the control seedlings. It was concluded that DNA polymorphisms observed with RAPD profile could be used as molecular marker for the evaluation of heavy metal induced genotoxic effects in plant species. The present results strongly suggested that Mentha arvensis could be used as a potential phytoremediator plant in mercury polluted environment. R. Manikandan, S. V. Sahi, and P. Venkatachalam Copyright © 2015 R. Manikandan et al. All rights reserved. Enhancing the Decolorizing and Degradation Ability of Bacterial Consortium Isolated from Textile Effluent Affected Area and Its Application on Seed Germination Sun, 11 Jan 2015 07:22:52 +0000 A bacterial consortium BMP1/SDSC/01 consisting of six isolates was isolated from textile effected soil, sludge, and textile effluent from Hudiara drain near Nishat Mills Limited, Ferozepur Road, Lahore, Pakistan. It was selected because of being capable of degrading and detoxifying red, green, black, and yellow textile dyes. The pH and supplements were optimized to enhance the decolorization ability of the selected consortium. The results indicated that decolorizing ability of consortium for the red, green, black, and yellow dyes was higher as compared to individual strains. The consortium was able to decolorize 84%, 84%, 85%, 85%, and 82% of 200 ppm of red, green, black, yellow, and mixed dyes within 24 h while individual strain required 72 h. On supplementing urea, the consortium decolorized 87, 86, 89, 86, and 83%, respectively, while on supplementing sodium chloride the consortium decolorized 93, 94, 93, 94, and 89% of red, green, black, yellow, and mixed dyes, respectively, which was maximum while in the presence of ascorbic acid and ammonium chloride it showed intermediate results. The effect of untreated and treated dyes was investigated on Zea mays L. (maize) and Sorghum vulgare Pers. (sorghum). This study will help to promote an efficient biotreatment of textile effluents. Rashid Mahmood, Faiza Sharif, Sikander Ali, and Muhammad Umar Hayyat Copyright © 2015 Rashid Mahmood et al. All rights reserved. Winter Wheat Water Productivity Evaluated by the Developed Remote Sensing Evapotranspiration Model in Hebei Plain, China Tue, 06 Jan 2015 07:58:39 +0000 Agricultural water is the main reason for the rapid decline of the NCP groundwater levels. It is of vital importance for the NCP sustainable agricultural development to master the ETa and its CWP. In this paper, the EBEM model was developed according to the theory of energy balance. From 2001 to 2006, the winter wheat ETa and CWP were estimated, and the spatial and temporal variations and their influencing factors were studied in the Hebei Plain. The results indicate that the EBEM model performed well by applying MODIS data to estimate the daily net radiation and ETa. For the daytime net radiation, the relative error between the estimation and the measurement amounted to 8.2% and the SEE was 0.82 MJ m−2/day. The average ETa deviation between the estimates and the measures amounted to 0.86 mm daily, and the SEE was 1.2 mm. The spatial variations indicated that the major distribution of ETa ranged from 350 to 450 mm, which trended downward within the study area from west to east. In the study period, the winter wheat CWP was mainly distributed between 0.29 and 1.67 kg/m3. In space, the CWP was higher in the west than in the east. Shengwei Zhang, Hongbin Zhao, Huimin Lei, Hongbo Shao, and Tingxi Liu Copyright © 2015 Shengwei Zhang et al. All rights reserved. Silicon Improves Maize Photosynthesis in Saline-Alkaline Soils Mon, 05 Jan 2015 13:05:49 +0000 The research aimed to determine the effects of Si application on photosynthetic characteristics of maize on saline-alkaline soil, including photosynthetic rate (), stomatal conductance (), transpiration rate (E), and intercellular CO2 concentration () of maize in the field with five levels (0, 45, 90, 150, and 225 kg·ha−1) of Si supplying. Experimental results showed that the values of , , and of maize were significantly enhanced while the values of E of maize were dramatically decreased by certain doses of silicon fertilizers, which meant that Si application with proper doses significantly increased photosynthetic efficiency of maize in different growth stages under stressing environment of saline-alkaline soil. The optimal dose of Si application in this experiment was 150 kg·ha−1 Si. It indicated that increase in maize photosynthesis under saline-alkaline stress took place by Si application with proper doses, which is helpful to improve growth and yield of maize. Zhiming Xie, Ri Song, Hongbo Shao, Fengbin Song, Hongwen Xu, and Yan Lu Copyright © 2015 Zhiming Xie et al. All rights reserved. Plant Abio-Stress and Bioresources Utilization for Sustainable Development Wed, 31 Dec 2014 11:28:28 +0000 Hong-bo Shao, Marian Brestic, Si-Xue Chen, Zhao Chang-Xing, and Xu Gang Copyright © 2014 Hong-bo Shao et al. All rights reserved. A Sequential Statistical Approach towards an Optimized Production of a Broad Spectrum Bacteriocin Substance from a Soil Bacterium Bacillus sp. YAS 1 Strain Mon, 29 Dec 2014 09:57:09 +0000 Bacteriocins, ribosomally synthesized antimicrobial peptides, display potential applications in agriculture, medicine, and industry. The present study highlights integral statistical optimization and partial characterization of a bacteriocin substance from a soil bacterium taxonomically affiliated as Bacillus sp. YAS 1 after biochemical and molecular identifications. A sequential statistical approach (Plackett-Burman and Box-Behnken) was employed to optimize bacteriocin (BAC YAS 1) production. Using optimal levels of three key determinants (yeast extract (0.48% (w/v), incubation time (62 hrs), and agitation speed (207 rpm)) in peptone yeast beef based production medium resulted in 1.6-fold enhancement in BAC YAS 1 level (470 AU/mL arbitrary units against Erwinia amylovora). BAC YAS 1 showed activity over a wide range of pH (1–13) and temperature (45–80°C). A wide spectrum antimicrobial activity of BAC YAS 1 against the human pathogens (Clostridium perfringens, Staphylococcus epidermidis, Campylobacter jejuni, Enterobacter aerogenes, Enterococcus sp., Proteus sp., Klebsiella sp., and Salmonella typhimurium), the plant pathogen (E. amylovora), and the food spoiler (Listeria innocua) was demonstrated. On top and above, BAC YAS 1 showed no antimicrobial activity towards lactic acid bacteria (Lactobacillus bulgaricus, L. casei, L. lactis, and L. reuteri). Promising characteristics of BAC YAS 1 prompt its commercialization for efficient utilization in several industries. Amira M. Embaby, Yasmin Heshmat, Ahmed Hussein, and Heba S. Marey Copyright © 2014 Amira M. Embaby et al. All rights reserved. rre37 Overexpression Alters Gene Expression Related to the Tricarboxylic Acid Cycle and Pyruvate Metabolism in Synechocystis sp. PCC 6803 Sun, 28 Dec 2014 13:41:42 +0000 The tricarboxylic acid (TCA) cycle and pyruvate metabolism of cyanobacteria are unique and important from the perspectives of biology and biotechnology research. Rre37, a response regulator induced by nitrogen depletion, activates gene expression related to sugar catabolism. Our previous microarray analysis has suggested that Rre37 controls the transcription of genes involved in sugar catabolism, pyruvate metabolism, and the TCA cycle. In this study, quantitative real-time PCR was used to measure the transcript levels of 12 TCA cycle genes and 13 pyruvate metabolism genes. The transcripts of 6 genes (acnB, icd, ppc, pyk1, me, and pta) increased after 4 h of nitrogen depletion in the wild-type GT strain but the induction was abolished by rre37 overexpression. The repression of gene expression of fumC, ddh, and ackA caused by nitrogen depletion was abolished by rre37 overexpression. The expression of me was differently affected by rre37 overexpression, compared to the other 24 genes. These results indicate that Rre37 differently controls the genes of the TCA cycle and pyruvate metabolism, implying the key reaction of the primary in this unicellular cyanobacterium. Hiroko Iijima, Atsuko Watanabe, Junko Takanobu, Masami Yokota Hirai, and Takashi Osanai Copyright © 2014 Hiroko Iijima et al. All rights reserved. Biochemical and Physiological Studies on the Effects of Senescence Leaves of Populus deltoides on Triticum vulgare Wed, 24 Dec 2014 11:22:57 +0000 Triticum vulgare (Wheat) based products are the major dietary source of food in developing countries. In India, it grows in association with boundary plantations of Populus deltoids (poplar). During winter, poplar enters in dormancy which cause a heavy leaf fall at the time of wheat seed germination. Large number of poplar senescence leaves may adversely affect the wheat. Therefore, the present study was performed to examine the effect of senescence poplar leaves on wheat germ and some other biochemical parameters. Seed’s germination rate was determined by measuring root and shoot lengths, percent germination, germination index, and inhibition percentage. Biochemical parameters, namely, pigment, carbohydrate, protein, and phenol content, were estimated. Activities of catalase and polyphenol oxidase which are stress marker enzymes were also measured. Results revealed that germination and other biochemical parameters of wheat were severely affected by senescence poplar leaves even at very low concentration. So, intercropping of poplar along with wheat may be chosen carefully as wheat is the major dietary staple. Tejinder Pal Khaket, Viney Kumar, Jasbir Singh, and Suman Dhanda Copyright © 2014 Tejinder Pal Khaket et al. All rights reserved. Effect of the Carbon Concentration, Blend Concentration, and Renewal Rate in the Growth Kinetic of Chlorella sp. Mon, 15 Dec 2014 09:47:26 +0000 The microalgae cultivation can be used as alternative sources of food, in agriculture, residual water treatment, and biofuels production. Semicontinuous cultivation is little studied but is more cost-effective than the discontinuous (batch) cultivation. In the semicontinuous cultivation, the microalga is maintained in better concentration of nutrients and the photoinhibition by excessive cell is reduced. Thus, biomass productivity and biocompounds of interest, such as lipid productivity, may be higher than in batch cultivation. The objective of this study was to examine the influence of blend concentration, medium renewal rate, and concentration of sodium bicarbonate on the growth of Chlorella sp. during semicontinuous cultivation. The cultivation was carried out in Raceway type bioreactors of 6 L, for 40 d at 30°C, 41.6 µmol m−2 s−1, and a 12 h light/dark photoperiod. Maximum specific growth rate (0.149 d−1) and generating biomass (2.89 g L−1) were obtained when the blend concentration was 0.80 g L−1, the medium renewal rate was 40%, and NaHCO3 was 1.60 g L−1. The average productivity (0.091 g L−1 d−1) was achieved with 0.8 g L−1 of blend concentration and NaHCO3 concentration of 1.6 g L−1, independent of the medium renewal rate. Adriano Arruda Henrard, Gabriel Martins da Rosa, Luiza Moraes, Michele Greque de Morais, and Jorge Alberto Vieira Costa Copyright © 2014 Adriano Arruda Henrard et al. All rights reserved. Production, Characterization of Tannase from Penicillium montanense URM 6286 under SSF Using Agroindustrial Wastes, and Application in the Clarification of Grape Juice (Vitis vinifera L.) Sun, 23 Nov 2014 12:57:29 +0000 Tannase is an enzyme that hydrolyzes esters and lateral bonds of tannins, such as tannic acid, releasing glucose and gallic acid and stands out in the clarification of wines and juices. Fungi of the genera Aspergillus and Penicillium are excellent producers of this enzyme. The search for fungi that produce high levels of tannase as well as new substrates for the enzyme production by the SSF is required. The objectives of this study were to evaluate the production of tannase by Aspergillus and Penicillium species through SSF using leaves and agroindustrial waste barbados cherry and mangaba fruit as substrate, select the best producer, optimize production, characterize the crude enzyme extract, and apply it the clarification of grape juice. Selecting the best producer was performed by planning Placket-Burman and RSM. P. montanense showed highest activity with 41.64 U/mL after 72 h of fermentation residue using barbados cherry, with 3.5% tannic acid and 70% moisture. The enzyme showed the highest activity at pH 9.0 and 50°C. The tannase of P. montanense was stable over a wide pH range and temperature and, when applied to grape juice, showed higher efficiency by reducing 46% of the tannin content after incubation 120 m. Juliana Silva de Lima, Roberta Cruz, Julyanna Cordoville Fonseca, Erika Valente de Medeiros, Marília de Holanda Cavalcanti Maciel, Keila Aparecida Moreira, and Cristina Maria de Souza Motta Copyright © 2014 Juliana Silva de Lima et al. All rights reserved. Saccharomyces cerevisiae Mixed Culture of Blackberry (Rubus ulmifolius L.) Juice: Synergism in the Aroma Compounds Production Sun, 23 Nov 2014 12:46:29 +0000 Blackberry (Rubus sp.) juice was fermented using four different strains of Saccharomyces cerevisiae (Vitilevure-CM4457, Enoferm-T306, ICV-K1, and Greroche Rhona-L3574) recognized because of their use in the wine industry. A medium alcoholic graduation spirit (<6) with potential to be produced at an industrial scale was obtained. Alcoholic fermentations were performed at , 200 rpm, and noncontrolled pH. The synergistic effect on the aromatic compounds production during fermentation in mixed culture was compared with those obtained by monoculture and physic mixture of spirits produced in monoculture. The aromatic composition was determined by HS-SPME-GC. The differences in aromatic profile principally rely on the proportions in aromatic compounds and not on the number of those compounds. The multivariance analysis, principal component analysis (PCA), and factorial discriminant analysis (DFA) permit to demonstrate the synergism between the strains. Pedro Ulises Bautista-Rosales, Juan Arturo Ragazzo-Sánchez, Gabriela Ruiz-Montañez, Rosa Isela Ortiz-Basurto, Guadalupe Luna-Solano, and Montserrat Calderón-Santoyo Copyright © 2014 Pedro Ulises Bautista-Rosales et al. All rights reserved. Evaluation of Selected Honey and One of Its Phenolic Constituent Eugenol against L1210 Lymphoid Leukemia Sun, 23 Nov 2014 07:36:36 +0000 People affected with leukemia are on the rise and several strategies were employed to thwart this deadly disease. Recent decade of research focuses on phenolic constituents as a tool for combating various inflammatory, cancer, and cardiac diseases. Our research showed honey and its phenolic constituents as crusaders against cancer. In this work, we explored the antileukemic activity of selected honey and one of its phenolic constituent eugenol against L1210 leukemia animal model. Results of this experiment showed that the selected honey samples as well as eugenol after intraperitoneal injection could not increase the median survival time (MST) of animals. Further, there was only slight marginal increase in the %T/C values of honey and eugenol treated groups. The number of phenolics present in the honey may not be a prime factor to promote antileukemic effect since there was no difference in the MST of two different honeys tested. This study limits the use of selected honey and eugenol against leukemia animal model. Saravana Kumar Jaganathan, Dilip Mondhe, Z. A. Wani, and Eko Supriyanto Copyright © 2014 Saravana Kumar Jaganathan et al. All rights reserved. Erratum to “Next Generation Sequencing: Potential and Application in Drug Discovery” Wed, 19 Nov 2014 13:11:34 +0000 Navneet Kumar Yadav, Pooja Shukla, Ankur Omer, Shruti Pareek, A. K. Srivastava, F. W. Bansode, and R. K. Singh Copyright © 2014 Navneet Kumar Yadav et al. All rights reserved. Overexpression of Bacterial mtlD Gene in Peanut Improves Drought Tolerance through Accumulation of Mannitol Tue, 11 Nov 2014 06:36:49 +0000 In the changing global environmental scenarios, water scarcity and recurrent drought impose huge reductions to the peanut (Arachis hypogaea L.) crop yield. In plants, osmotic adjustments associated with efficient free radical scavenging ability during abiotic stress are important components of stress tolerance mechanisms. Mannitol, a compatible solute, is known to scavenge hydroxyl radicals generated during various abiotic stresses, thereby conferring tolerance to water-deficit stress in many plant species. However, peanut plant is not known to synthesize mannitol. Therefore, bacterial mtlD gene coding for mannitol 1-phosphate dehydrogenase under the control of constitutive promoter CaMV35S was introduced and overexpressed in the peanut cv. GG 20 using Agrobacterium tumefaciens-mediated transformation. A total of eight independent transgenic events were confirmed at molecular level by PCR, Southern blotting, and RT-PCR. Transgenic lines had increased amount of mannitol and exhibited enhanced tolerance in response to water-deficit stress. Improved performance of the mtlD transgenics was indicated by excised-leaf water loss assay and relative water content under water-deficit stress. Better performance of transgenics was due to the ability of the plants to synthesize mannitol. However, regulation of mtlD gene expression in transgenic plants remains to be elucidated. Tengale Dipak Bhauso, Thankappan Radhakrishnan, Abhay Kumar, Gyan Prakash Mishra, Jentilal Ramjibhai Dobaria, Kirankumar Patel, and Manchikatla Venkat Rajam Copyright © 2014 Tengale Dipak Bhauso et al. All rights reserved. Zinc Oxide Nanoparticles for Revolutionizing Agriculture: Synthesis and Applications Tue, 11 Nov 2014 00:00:00 +0000 Nanotechnology is the most innovative field of 21st century. Extensive research is going on for commercializing nanoproducts throughout the world. Due to their unique properties, nanoparticles have gained considerable importance compared to bulk counterparts. Among other metal nanoparticles, zinc oxide nanoparticles are very much important due to their utilization in gas sensors, biosensors, cosmetics, drug-delivery systems, and so forth. Zinc oxide nanoparticles (ZnO NPs) also have remarkable optical, physical, and antimicrobial properties and therefore have great potential to enhance agriculture. As far as method of formation is concerned, ZnO NPs can be synthesized by several chemical methods such as precipitation method, vapor transport method, and hydrothermal process. The biogenic synthesis of ZnO NPs by using different plant extracts is also common nowadays. This green synthesis is quite safe and ecofriendly compared to chemical synthesis. This paper elaborates the synthesis, properties, and applications of zinc oxide nanoparticles. Sidra Sabir, Muhammad Arshad, and Sunbal Khalil Chaudhari Copyright © 2014 Sidra Sabir et al. All rights reserved. A Lectin from Dioclea violacea Interacts with Midgut Surface of Lutzomyia migonei, Unlike Its Homologues, Cratylia floribunda Lectin and Canavalia gladiata Lectin Wed, 05 Nov 2014 10:37:13 +0000 Leishmaniasis is a vector-borne disease transmitted by phlebotomine sand fly. Susceptibility and refractoriness to Leishmania depend on the outcome of multiple interactions that take place within the sand fly gut. Promastigote attachment to sand fly midgut epithelium is essential to avoid being excreted together with the digested blood meal. Promastigote and gut sand fly surface glycans are important ligands in this attachment. The purpose of the present study was to evaluate the interaction of three lectins isolated from leguminous seeds (Diocleinae subtribe), D-glucose and D-mannose-binding, with glycans on Lutzomyia migonei midgut. To study this interaction the lectins were labeled with FITC and a fluorescence assay was performed. The results showed that only Dioclea violacea lectin (DVL) was able to interact with midgut glycans, unlike Cratylia floribunda lectin (CFL) and Canavalia gladiata lectin (CGL). Furthermore, when DVL was blocked with D-mannose the interaction was inhibited. Differences of spatial arrangement of residues and volume of carbohydrate recognition domain (CRD) may be the cause of the fine specificity of DVL for glycans in the surface on Lu. migonei midgut. The findings in this study showed the presence of glycans in the midgut with glucose/mannose residues in its composition and these residues may be important in interaction between Lu. migonei midgut and Leishmania. Juliana Montezuma Barbosa Monteiro Tínel, Melina Fechine Costa Benevides, Mércia Sindeaux Frutuoso, Camila Farias Rocha, Francisco Vassiliepe Sousa Arruda, Mayron Alves Vasconcelos, Francisco Nascimento Pereira-Junior, João Batista Cajazeiras, Kyria Santiago do Nascimento, Jorge Luiz Martins, Edson Holanda Teixeira, Benildo Sousa Cavada, Ricardo Pires dos Santos, and Margarida Maria Lima Pompeu Copyright © 2014 Juliana Montezuma Barbosa Monteiro Tínel et al. All rights reserved. Identification and Validation of a New Male Sex-Specific ISSR Marker in Pointed Gourd (Trichosanthes dioica Roxb.) Wed, 05 Nov 2014 00:00:00 +0000 The aim of the present study was to develop a genetic sex marker for the pointed gourd (Trichosanthes dioica Roxb.) to allow gender determination at any stage in the life cycle. Screening of genomic DNA with intersimple sequence repeat (ISSR) primers was used to discover sex-specific touch-down polymerase chain reaction (Td-PCR) amplification products. Using pooled DNA from male and female genotypes and 42 ISSR primers, a putative male specific marker (~550 bp) was identified. DNA marker specific to male is an indication of existence of nonepigenetic factors involved in gender development in pointed gourd. The ISSR technique has proved to be a reliable technique in gender determination of pointed gourd genotypes at the seedling phenophase. The sex marker developed here could also be used as a starting material towards sequence characterization of sex linked genes for better understanding the developmental as well as evolutionary pathways in sexual dimorphism. Sinchan Adhikari, Soumen Saha, Tapas Kumar Bandyopadhyay, and Parthadeb Ghosh Copyright © 2014 Sinchan Adhikari et al. All rights reserved. The Assessment of Cholinesterase from the Liver of Puntius Javanicus as Detection of Metal Ions Mon, 27 Oct 2014 06:31:36 +0000 Crude extract of ChE from the liver of Puntius javanicus was purified using procainamide-sepharyl 6B. S-Butyrylthiocholine iodide (BTC) was selected as the specific synthetic substrate for this assay with the highest maximal velocity and lowest biomolecular constant at 53.49 µmole/min/mg and 0.23 mM, respectively, with catalytic efficiency ratio of 0.23. The optimum parameter was obtained at pH 7.5 and optimal temperature in the range of 25 to 30°C. The effect of different storage condition was assessed where ChE activity was significantly decreased after 9 days of storage at room temperature. However, ChE activity showed no significant difference when stored at 4.0, 0, and −25°C for 15 days. Screening of heavy metals shows that chromium, copper, and mercury strongly inhibited P. javanicus ChE by lowering the activity below 50%, while several pairwise combination of metal ions exhibited synergistic inhibiting effects on the enzyme which is greater than single exposure especially chromium, copper, and mercury. The results showed that P. javanicus ChE has the potential to be used as a biosensor for the detection of metal ions. Mohd Khalizan Sabullah, Mohd Rosni Sulaiman, Mohd Yunus Abd Shukor, Mohd Arif Syed, Nor Aripin Shamaan, Ariff Khalid, and Siti Aqlima Ahmad Copyright © 2014 Mohd Khalizan Sabullah et al. All rights reserved. Molecular Markers and Cotton Genetic Improvement: Current Status and Future Prospects Thu, 23 Oct 2014 12:59:30 +0000 Narrow genetic base and complex allotetraploid genome of cotton (Gossypium hirsutum L.) is stimulating efforts to avail required polymorphism for marker based breeding. The availability of draft genome sequence of G. raimondii and G. arboreum and next generation sequencing (NGS) technologies facilitated the development of high-throughput marker technologies in cotton. The concepts of genetic diversity, QTL mapping, and marker assisted selection (MAS) are evolving into more efficient concepts of linkage disequilibrium, association mapping, and genomic selection, respectively. The objective of the current review is to analyze the pace of evolution in the molecular marker technologies in cotton during the last ten years into the following four areas: (i) comparative analysis of low- and high-throughput marker technologies available in cotton, (ii) genetic diversity in the available wild and improved gene pools of cotton, (iii) identification of the genomic regions within cotton genome underlying economic traits, and (iv) marker based selection methodologies. Moreover, the applications of marker technologies to enhance the breeding efficiency in cotton are also summarized. Aforementioned genomic technologies and the integration of several other omics resources are expected to enhance the cotton productivity and meet the global fiber quantity and quality demands. Waqas Malik, Javaria Ashraf, Muhammad Zaffar Iqbal, Asif Ali Khan, Abdul Qayyum, Muhammad Ali Abid, Etrat Noor, Muhammad Qadir Ahmad, and Ghulam Hasan Abbasi Copyright © 2014 Waqas Malik et al. All rights reserved. Biobleaching of Industrial Important Dyes with Peroxidase Partially Purified from Garlic Thu, 23 Oct 2014 11:42:38 +0000 An acidic peroxidase was extracted from garlic (Allium sativum) and was partially purified threefold by ammonium sulphate precipitation, dialysis, and gel filtration chromatography using sephadex G-200. The specific activity of the enzyme increased from 4.89 U/mg after ammonium sulphate precipitation to 25.26 U/mg after gel filtration chromatography. The optimum temperature and pH of the enzyme were 50°C and 5.0, respectively. The Km and for H2O2 and o-dianisidine were 0.026 mM and 0.8 U/min, and 25 mM and 0.75 U/min, respectively. Peroxidase from garlic was effective in decolourizing Vat Yellow 2, Vat Orange 11, and Vat Black 27 better than Vat Green 9 dye. For all the parameters monitored, the decolourization was more effective at a pH range, temperature, H2O2 concentration, and enzyme concentration of 4.5–5.0, 50°C, 0.6 mM, and 0.20 U/mL, respectively. The observed properties of the enzyme together with its low cost of extraction (from local sources) show the potential of this enzyme for practical application in industrial wastewater treatment especially with hydrogen peroxide. These Vat dyes also exhibited potentials of acting as peroxidase inhibitors at alkaline pH range. Akudo Chigozirim Osuji, Sabinus Oscar O. Eze, Emmanuel Emeka Osayi, and Ferdinand Chiemeka Chilaka Copyright © 2014 Akudo Chigozirim Osuji et al. All rights reserved. Effect of Continuous Cropping Generations on Each Component Biomass of Poplar Seedlings during Different Growth Periods Thu, 23 Oct 2014 07:14:38 +0000 In order to investigate the change rules and response characteristics of growth status on each component of poplar seedling followed by continuous cropping generations and growth period, we clear the biomass distribution pattern of poplar seedling, adapt continuous cropping, and provide theoretical foundation and technical reference on cultivation management of poplar seedling, the first generation, second generation, and third generation continuous cropping poplar seedlings were taken as study objects, and the whole poplar seedling was harvested to measure and analyze the change of each component biomass on different growth period poplar leaves, newly emerging branches, trunks and root system, and so forth. The results showed that the whole biomass of poplar seedling decreased significantly with the leaf area and its ratio increased, and the growth was inhibited obviously. The biomass aboveground was more than that underground. The ratios of leaf biomass and newly emerging branches biomass of first continuous cropping poplar seedling were relatively high. With the continuous cropping generations and growth cycle increasing, poplar seedling had a growth strategy to improve the ratio of root-shoot and root-leaf to adapt the limited soil nutrient of continuous cropping. Jiangbao Xia, Shuyong Zhang, Tian Li, Xia Liu, Ronghua Zhang, and Guangcan Zhang Copyright © 2014 Jiangbao Xia et al. All rights reserved. In Vitro Pollen Viability and Pollen Germination in Cherry Laurel (Prunus laurocerasus L.) Wed, 22 Oct 2014 00:00:00 +0000 Pollen quality is important for growers and breeders. This study was carried out to determine in vitro pollen viability and pollen germination in seven genotypes of cherry laurel (Prunus laurocerasus L.). Two pollen viability tests, TTC (2,3,5-triphenyl tetrazolium chloride) and IKI (iodine potassium iodide), were used. Pollen traits of genotypes were studied using an in vitro medium containing 0%, 5%, 10%, 15%, and 20% sucrose to determine the best sucrose concentrations for germination. In the second step, the germinated pollen was counted 1, 4, 6, 10, 12, 24, and 48 hours later until there was no further germination. The viability rates were different according to genotypes and tests used. The IKI and TTC staining tests and pollen germination had low correlation ( = 0.0614 and = 0.0015, resp.). Painted pollen rate was higher and pollen was well-stained with IKI test and pollen viability estimated with TTC staining test was better than that estimated with the IKI staining test. 15% sucrose gave the best germination rates in most of the genotypes. Pollen germination rates were recorded periodically from one hour to 48 hours in 15% sucrose and the results showed that pollen germination rates increased after 6 hours of being placed in culture media. Melekber Sulusoglu and Aysun Cavusoglu Copyright © 2014 Melekber Sulusoglu and Aysun Cavusoglu. All rights reserved. Optimization of Biosorptive Removal of Dye from Aqueous System by Cone Shell of Calabrian Pine Tue, 21 Oct 2014 09:50:12 +0000 The biosorption performance of raw cone shell of Calabrian pine for C.I. Basic Red 46 as a model azo dye from aqueous system was optimized using Taguchi experimental design methodology. L9 (33) orthogonal array was used to optimize the dye biosorption by the pine cone shell. The selected factors and their levels were biosorbent particle size, dye concentration, and contact time. The predicted dye biosorption capacity for the pine cone shell from Taguchi design was obtained as 71.770 mg g−1 under optimized biosorption conditions. This experimental design provided reasonable predictive performance of dye biosorption by the biosorbent (: 0.9961). Langmuir model fitted better to the biosorption equilibrium data than Freundlich model. This displayed the monolayer coverage of dye molecules on the biosorbent surface. Dubinin-Radushkevich model and the standard Gibbs free energy change proposed physical biosorption for predominant mechanism. The logistic function presented the best fit to the data of biosorption kinetics. The kinetic parameters reflecting biosorption performance were also evaluated. The optimization study revealed that the pine cone shell can be an effective and economically feasible biosorbent for the removal of dye. Fatih Deniz Copyright © 2014 Fatih Deniz. All rights reserved. Establishment of an Efficient In Vitro Regeneration Protocol for Rapid and Mass Propagation of Dendrobium chrysotoxum Lindl. Using Seed Culture Mon, 20 Oct 2014 13:40:50 +0000 An efficient in vitro regeneration protocol from seed culture has been established successfully for Dendrobium chrysotoxum, an epiphytic orchid having tremendous ornamental and medicinal values. Seed germination response was encouraging in Mitra (M) medium enriched with different combinations of auxins and cytokinins. Medium supplemented with 0.4% activated charcoal (AC), 2 mg/L 6-benzyl amino purine (BAP), and 2 mg/L indole-3-acetic acid (IAA) produced best seed germination percentage in 2 weeks of culture. Incorporation of higher concentration of kinetin (KN) or BAP in combination with low auxin in medium induced pronounced shooting and leaf formation. Reduction in leaf development was evident when cytokinins exist singly in medium indicating synergistic effect of auxin and cytokinin in leaf induction. Presence of elevated level of indole-3-butyric acid (IBA) or 1-naphthalene acetic acid (NAA) with low cytokinin content in medium generated more in vitro rooting, though IBA was found to be more effective in rooting induction as compared to NAA. The in vitro protocol for asymbiotic seed germination developed from the present investigation can be used for rapid mass propagation of this highly important Dendrobium orchid species. Potshangbam Nongdam and Leimapokpam Tikendra Copyright © 2014 Potshangbam Nongdam and Leimapokpam Tikendra. All rights reserved. Response Surface Methodology: Optimisation of Antifungal Bioemulsifier from Novel Bacillus thuringiensis Tue, 14 Oct 2014 07:01:15 +0000 An antifungal bioemulsifier compound was produced from a novel strain of Bacillus thuringiensis pak2310. To accentuate the production and as the first step to improve the yield, a central composite design (CCD) was used to study the effect of various factors like minimal salts (1X and 3X), glycerol concentration (2% and 4%), beef extract concentration (1% and 3%), and sunflower oil concentration (2% and 4%) on the production of bioemulsifier molecule and to optimize the conditions to increase the production. The emulsification index was used as the response variable as the increase in surfactant production was seen to be proportional to increased emulsification. A quadratic equation was employed to express the response variable in terms of the independent variables. Statistical tools like student’s -test, -test, and ANOVA were employed to identify the important factors and to test the adequacy of the model. Under optimum conditions (1X concentration of minimal salts (MS), 2.6% glycerol (v/v), 1% beef extract (w/v), and 2% sunflower oil (v/v)) a 65% increase in yield was produced. Deepak Rajendran, Ponnusami Venkatachalam, and Jayapradha Ramakrishnan Copyright © 2014 Deepak Rajendran et al. All rights reserved. Ethylene Response Factor Sl-ERF.B.3 Is Responsive to Abiotic Stresses and Mediates Salt and Cold Stress Response Regulation in Tomato Wed, 06 Aug 2014 07:47:43 +0000 Sl-ERF.B.3 (Solanum lycopersicum ethylene response factor B.3) gene encodes for a tomato transcription factor of the ERF (ethylene responsive factor) family. Our results of real-time RT-PCR showed that Sl-ERF.B.3 is an abiotic stress responsive gene, which is induced by cold, heat, and flooding, but downregulated by salinity and drought. To get more insight into the role of Sl-ERF.B.3 in plant response to separate salinity and cold, a comparative study between wild type and two Sl-ERF.B.3 antisense transgenic tomato lines was achieved. Compared with wild type, Sl-ERF.B.3 antisense transgenic plants exhibited a salt stress dependent growth inhibition. This inhibition was significantly enhanced in shoots but reduced in roots, leading to an increased root to shoot ratio. Furthermore, the cold stress essay clearly revealed that introducing antisense Sl-ERF.B.3 in transgenic tomato plants reduces their cell injury and enhances their tolerance against 14 d of cold stress. All these results suggest that Sl-ERF.B.3 gene is involved in plant response to abiotic stresses and may play a role in the layout of stress symptoms under cold stress and in growth regulation under salinity. Imen Klay, Julien Pirrello, Leila Riahi, Anne Bernadac, Ameur Cherif, Mondher Bouzayen, and Sadok Bouzid Copyright © 2014 Imen Klay et al. All rights reserved. An Integrated Use of Topography with RSI in Gully Mapping, Shandong Peninsula, China Tue, 05 Aug 2014 07:38:17 +0000 Taking the Quickbird optical satellite imagery of the small watershed of Beiyanzigou valley of Qixia city, Shandong province, as the study data, we proposed a new method by using a fused image of topography with remote sensing imagery (RSI) to achieve a high precision interpretation of gully edge lines. The technique first transformed remote sensing imagery into HSV color space from RGB color space. Then the slope threshold values of gully edge line and gully thalweg were gained through field survey and the slope data were segmented using thresholding, respectively. Based on the fused image in combination with gully thalweg thresholding vectors, the gully thalweg thresholding vectors were amended. Lastly, the gully edge line might be interpreted based on the amended gully thalweg vectors, fused image, gully edge line thresholding vectors, and slope data. A testing region was selected in the study area to assess the accuracy. Then accuracy assessment of the gully information interpreted by both interpreting remote sensing imagery only and the fused image was performed using the deviation, kappa coefficient, and overall accuracy of error matrix. Compared with interpreting remote sensing imagery only, the overall accuracy and kappa coefficient are increased by 24.080% and 264.364%, respectively. The average deviations of gully head and gully edge line are reduced by 60.448% and 67.406%, respectively. The test results show the thematic and the positional accuracy of gully interpreted by new method are significantly higher. Finally, the error sources for interpretation accuracy by the two methods were analyzed. Fuhong He, Tao Wang, Lijuan Gu, Tao Li, Weiguo Jiang, and Hongbo Shao Copyright © 2014 Fuhong He et al. All rights reserved. Analysis of Phenolic Acids of Jerusalem Artichoke (Helianthus tuberosus L.) Responding to Salt-Stress by Liquid Chromatography/Tandem Mass Spectrometry Tue, 05 Aug 2014 07:35:08 +0000 Plant phenolics can have applications in pharmaceutical and other industries. To identify and quantify the phenolic compounds in Helianthus tuberosus leaves, qualitative analysis was performed by a reversed phase high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) and quantitative analysis by HPLC. Ten chlorogenic acids (CGAs) were identified (3-o-caffeoylquinic acid, two isomers of caffeoylquinic acid, caffeic acid, p-coumaroyl-quinic acid, feruloylquinic acid, 3,4-dicaffeoyquinic acid, 3,5-dicaffeoylquinic acid, 1,5-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid) by comparing their retention times, UV-Vis absorption spectra, and MS/MS spectra with standards. In addition, four other phenolic compounds, including caffeoyl glucopyranose, isorhamnetin glucoside, kaempferol glucuronide, and kaempferol-3-o-glucoside, were tentatively identified in Helianthus tuberosus leaves for the first time. The 3-o-caffeoylquinic acid (7.752 mg/g DW), 4,5-dicaffeoylquinic acid (5.633 mg/g DW), and 3,5-dicaffeoylquinic acid (4.900 mg/g DW) were the major phenolic compounds in leaves of Helianthus tuberosus cultivar NanYu in maturity. The variations in phenolic concentrations and proportions in Helianthus tuberosus leaves were influenced by genotype and plant growth stage. Cultivar NanYu had the highest concentration of phenolic compounds, in particular 3-o-caffeoylquinic acid and 4,5-dicaffeoylquinic acid compared with the other genotypes (wild accession and QingYu). Considering various growth stages, the concentration of total phenolics in cultivar NanYu was higher at flowering stage (5.270 mg/g DW) than at budding and tuber swelling stages. Cultivar NanYu of Helianthus tuberosus is a potential source of natural phenolics that may play an important role in the development of pharmaceuticals. Fujia Chen, Xiaohua Long, Zhaopu Liu, Hongbo Shao, and Ling Liu Copyright © 2014 Fujia Chen et al. All rights reserved. Synthesized Attributes of Water Use by Regional Vegetation: A Key to Cognition of “Water Pump” Viewpoint Thu, 24 Jul 2014 08:32:46 +0000 Recently, the frequent seasonal drought in Southwest China has brought considerable concerns and continuous heated arguments on the “water pump” viewpoint (i.e., the water demand from Hevea spp. and Eucalyptus spp. can be treated as a water pump) once again. However, such viewpoint just focused on water consumption from vegetation transpiration and its ecoenvironment impacts, which had not considered other attributes of vegetation, namely, water saving and drought resistance, and hydrological regulation (water conservation) into consideration. Thus, in this paper, the synthesized attributes of regional vegetation water use had been mainly discussed. The results showed that the study on such aspects as the characters of water consumption from vegetation transpiration, the potential of water saving and drought resistance, and the effects of hydrological regulation in Southwest China lagged far behind, let alone the report on synthesized attributes of water utilization with the organic combination of the three aspects above or the paralleled analysis. Accordingly, in this paper, the study on the synthesized attributes of water use by regional vegetation in Southwest China was suggested, and the objectives of such a special study were clarified, targeting the following aspects: (i) characters of water consumption from transpiration of regional typical artificial vegetation; (ii) potential of water saving and drought resistance of regional typical artificial vegetation; (iii) effects of hydrological regulation of regional typical artificial vegetation; (iv) synthesized attributes of water use by regional typical artificial vegetation. It is expected to provide a new idea for the scientific assessment on the regional vegetation ecoenvironment effects and theoretical guidance for the regional vegetation reconstruction and ecological restoration. Xin-hui Huang, Fu-ke Yu, Xiao-ying Li, Yuan Zheng, Hua Yuan, Jian-gang Ma, Yan-xia Wang, Dan-hui Qi, and Hong-bo Shao Copyright © 2014 Xin-hui Huang et al. All rights reserved. Stimulatory Effects of Gamma Irradiation on Phytochemical Properties, Mitotic Behaviour, and Nutritional Composition of Sainfoin (Onobrychis viciifolia Scop.) Wed, 23 Jul 2014 00:00:00 +0000 Sainfoin (Onobrychis viciifolia Scop. Syn. Onobrychis sativa L.) is a bloat-safe forage crop with high levels of tannins, which is renowned for its medicinal qualities in grazing animals. Mutagenesis technique was applied to investigate the influence of gamma irradiation at 30, 60, 90, and 120 Gy on mitotic behavior, in vitro growth factors, phytochemical and nutritional constituents of sainfoin. Although a percentage of plant necrosis and non-growing seed were enhanced by irradiation increment, the germination speed was significantly decreased. It was observed that gamma irradiated seeds had higher value of crude protein and dry matter digestibility compared to control seeds. Toxicity of copper was reduced in sainfoin irradiated seeds at different doses of gamma rays. Anthocyanin content also decreased in inverse proportion to irradiation intensity. Accumulation of phenolic and flavonoid compounds was enhanced by gamma irradiation exposure in leaf cells. HPLC profiles differed in peak areas of the two important alkaloids, Berberine and Sanguinarine, in 120 Gy irradiated seeds compared to control seeds. There were positive correlations between irradiation dose and some abnormality divisions such as laggard chromosome, micronucleus, binucleated cells, chromosome bridge, and cytomixis. In reality, radiocytological evaluation was proven to be essential in deducing the effectiveness of gamma irradiation to induce somaclonal variation in sainfoin. Sadegh Mohajer, Rosna Mat Taha, Ma Ma Lay, Arash Khorasani Esmaeili, and Mahsa Khalili Copyright © 2014 Sadegh Mohajer et al. All rights reserved. Effects of Salt-Drought Stress on Growth and Physiobiochemical Characteristics of Tamarix chinensis Seedlings Tue, 22 Jul 2014 00:00:00 +0000 The present study was designed to clarify the effects of salinity and water intercross stresses on the growth and physiobiochemical characteristics of Tamarix chinensis seedlings by pots culture under the artificial simulated conditions. The growth, activities of SOD, POD, and contents of MDA and osmotic adjusting substances of three years old seedlings of T. chinensis were studied under different salt-drought intercross stress. Results showed that the influence of salt stress on growth was greater than drought stress, the oxidation resistance of SOD and POD weakened gradually with salt and drought stresses intensified, and the content of MDA was higher under severe drought and mild and moderate salt stresses. The proline contents increased with the stress intensified but only significantly higher than control under the intercross stresses of severe salt-severe drought. It implied that T. chinensis could improve its stress resistance by adjusted self-growth and physiobiochemical characteristics, and the intercross compatibility of T. chinensis to salt and drought stresses can enhance the salt resistance under appropriate drought stress, but the dominant factors influencing the physiological biochemical characteristics of T. chinensis were various with the changing of salt-drought intercross stresses gradients. Junhua Liu, Jiangbao Xia, Yanming Fang, Tian Li, and Jingtao Liu Copyright © 2014 Junhua Liu et al. All rights reserved. Micropropagation of an Exotic Ornamental Plant, Calathea crotalifera, for Production of High Quality Plantlets Sun, 20 Jul 2014 00:00:00 +0000 A successful protocol was established for micropropagation in two selected varieties of exotic ornamental plants, Calathea crotalifera. The effects of different sterilization techniques, explant type, and the combination and concentration of plant growth regulators on shoots induction were studied. The axillary shoot buds explants sprouted from rhizomes in soil free conditions showed high induction rate of shoots with lowest contamination percentage when treated with combination of 30% (v/v) NaOCl, 70% (v/v) ethanol, and 0.3% (w/v) HgCl2. In the present study, the highest number of multiple shoots was obtained in MS basal medium supplemented with 3.5 mg/L 6-Benzylaminopurine (BAP), 1.0 mg/L 1-Naphthaleneacetic acid (NAA), 3% sucrose, and 6 g/L plant agar for both varieties and was used as multiplication medium. Microshoots were highly induced when the young shoot bud explants were incised longitudinally prior subculture. Chlorophyll analysis was studied to test the effects of activated charcoal and L-glutamine on reduction of necrosis problem. The maximum roots induction was recorded on MS medium supplemented with 1.0 mg/L 1-Naphthaleneacetic acid (NAA) compared to indolebutyric acid (IBA). The complete regenerated plantlets were successfully acclimatized in the soilless medium under greenhouse condition. This is the first report of rapid mass propagation for C. crotalifera. Shahril Efzueni Rozali, Kamaludin A. Rashid, and Rosna Mat Taha Copyright © 2014 Shahril Efzueni Rozali et al. All rights reserved. Detoxification of Corncob Acid Hydrolysate with SAA Pretreatment and Xylitol Production by Immobilized Candida tropicalis Tue, 15 Jul 2014 00:00:00 +0000 Xylitol fermentation production from corncob acid hydrolysate has become an attractive and promising process. However, corncob acid hydrolysate cannot be directly used as fermentation substrate owing to various inhibitors. In this work, soaking in aqueous ammonia (SAA) pretreatment was employed to reduce the inhibitors in acid hydrolysate. After detoxification, the corncob acid hydrolysate was fermented by immobilized Candida tropicalis cell to produce xylitol. Results revealed that SAA pretreatment showed high delignification and efficient removal of acetyl group compounds without effect on cellulose and xylan content. Acetic acid was completely removed, and the content of phenolic compounds was reduced by 80%. Furthermore, kinetic behaviors of xylitol production by immobilized C. tropicalis cell were elucidated from corncob acid hydrolysate detoxified with SAA pretreatment and two-step adsorption method, respectively. The immobilized C. tropicalis cell showed higher productivity efficiency using the corncob acid hydrolysate as fermentation substrate after detoxification with SAA pretreatment than by two-step adsorption method in the five successive batch fermentation rounds. After the fifth round fermentation, about 60 g xylitol/L fermentation substrate was obtained for SAA pretreatment detoxification, while about 30 g xylitol/L fermentation substrate was obtained for two-step adsorption detoxification. Li-Hong Deng, Yong Tang, and Yun Liu Copyright © 2014 Li-Hong Deng et al. All rights reserved. Impacts of Groundwater Recharge from Rubber Dams on the Hydrogeological Environment in Luoyang Basin, China Mon, 14 Jul 2014 00:00:00 +0000 In the rubber dam’s impact area, the groundwater total hardness (TH) has declined since 2000, ultimately dropping to 100–300 mg/L in 2012. pH levels have shown no obvious changes. NH4-N concentration in the groundwater remained stable from 2000 to 2006, but it increased from 2007 to 2012, with the largest increase up to 0.2 mg/L. NO3-N concentration in the groundwater generally declined in 2000–2006 and then increased from 2007; the largest increase was to 10 mg/L in 2012. Total dissolved solids (TDS) of the groundwater showed a general trend of decline from 2000 to 2009, but levels increased after 2010, especially along the south bank of the Luohe River where the largest increase recorded was approximately 100 mg/L. This study has shown that the increases in the concentrations of NH4-N and NO3-N were probably caused by changes in groundwater levels. Nitrates adsorbed by the silt clay of aeration zone appear to have entered the groundwater through physical and chemical reactions. TDS increased because of groundwater evaporation and some soluble ions entered the groundwater in the unsaturated zone. The distance of the contaminant to the surface of the aquifer became shorter due to the shallow depth of groundwater, resulting in the observed rise in pollutant concentrations more pronounced. Shaogang Dong, Baiwei Liu, Huamin Liu, Shidong Wang, and Lixin Wang Copyright © 2014 Shaogang Dong et al. All rights reserved. Expression Profiles of 12 Late Embryogenesis Abundant Protein Genes from Tamarix hispida in Response to Abiotic Stress Thu, 10 Jul 2014 09:57:33 +0000 Twelve embryogenesis abundant protein (LEA) genes (named ThLEA-1 to -12) were cloned from Tamarix hispida. The expression profiles of these genes in response to NaCl, PEG, and abscisic acid (ABA) in roots, stems, and leaves of T. hispida were assessed using real-time reverse transcriptase-polymerase chain reaction (RT-PCR). These ThLEAs all showed tissue-specific expression patterns in roots, stems, and leaves under normal growth conditions. However, they shared a high similar expression patterns in the roots, stems, and leaves when exposed to NaCl and PEG stress. Furthermore, ThLEA-1, -2, -3, -4, and -11 were induced by NaCl and PEG, but ThLEA-5, -6, -8, -10, and -12 were downregulated by salt and drought stresses. Under ABA treatment, some ThLEA genes, such as ThLEA-1, -2, and -3, were only slightly differentially expressed in roots, stems, and leaves, indicating that they may be involved in the ABA-independent signaling pathway. These findings provide a basis for the elucidation of the function of LEA genes in future work. Caiqiu Gao, Yali Liu, Chao Wang, Kaimin Zhang, and Yucheng Wang Copyright © 2014 Caiqiu Gao et al. All rights reserved. Molecular Cloning and Bioinformatics Analysis of a New Plasma Membrane Na+/H+ Antiporter Gene from the Halophyte Kosteletzkya virginica Mon, 30 Jun 2014 12:05:28 +0000 A new plasma membrane Na+/H+ antiporter gene (named as KvSOS1) was cloned from the halophyte Kosteletzkya virginica by reverse-transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) technology, which is a homologue of SOS1 (salt overly sensitive 1). The full-length cDNA is 3850 bp and contains an open reading frame (ORF) encoding a protein of 1147 amino acids with a molecular weight of 127.56 kDa and a theoretical pI of 6.18. Bioinformatics analysis indicated that the deduced protein appears to be a transmembrane protein with 12 transmembrane domains at the N-terminal region and a long hydrophilic tail in cytoplasm at its C-terminal region and shares 72–82% identity at the peptide level with other plant plasma membrane Na+/H+ antiporters. Hongyan Wang, Xiaoli Tang, Chuyang Shao, Hongbo Shao, and Honglei Wang Copyright © 2014 Hongyan Wang et al. All rights reserved. Responses of Seed Germination, Seedling Growth, and Seed Yield Traits to Seed Pretreatment in Maize (Zea mays L.) Thu, 26 Jun 2014 08:50:59 +0000 A series of seed priming experiments were conducted to test the effects of different pretreatment methods to seed germination, seedling growth, and seed yield traits in maize (Zea mays L.). Results indicated that the seeds primed by gibberellins (GA), NaCl, and polyethylene glycol (PEG) reagents showed a higher imbibitions rate compared to those primed with water. The final germination percentage and germination rate varied with different reagents significantly (). The recommended prime reagents were GA at 10 mg/L, NaCl at 50 mM, and PEG at 15% on account of germination experiment. 15% PEG priming reagent increased shoot and root biomass of maize seedling. The shoot biomass of seedlings after presoaking the seeds with NaCl reagent was significantly higher than the seedlings without priming treatment. No significant differences of plant height, leaf number, and hundred-grain weight were observed between control group and priming treatments. Presoaking with water, NaCl (50 mM), or PEG (15%) significantly increased the hundred-grain weight of maize. Therefore, seed pretreatment is proved to be an effective technique to improve the germination performance, seedling growth, and seed yield of maize. However, when compared with the two methods, if immediate sowing is possible, presoaking is recommended to harvest better benefits compared to priming method. Yu Tian, Bo Guan, Daowei Zhou, Junbao Yu, Guangdi Li, and Yujie Lou Copyright © 2014 Yu Tian et al. All rights reserved. Production and Purification of a Novel Xanthan Lyase from a Xanthan-Degrading Microbacterium sp. Strain XT11 Thu, 26 Jun 2014 08:46:51 +0000 A xanthan lyase was produced and purified from the culture supernatant of an excellent xanthan-modifying strain Microbacterium sp. XT11. Xanthan lyase was induced by xanthan but was inhibited by its structural monomer glucose. Its production by strain XT11 is much higher than that by all other reported strains. The purified xanthan lyase has a molecular mass of 110 kDa and a specific activity of 28.2 U/mg that was much higher than that of both Paenibacillus and Bacillus lyases. It was specific on the pyruvated mannosyl residue in the intact xanthan molecule, but about 50% lyase activity remained when xanthan was partially depyruvated. Xanthan lyase was optimally active at pH 6.0–6.5 and 40°C and alkali-tolerant at a high pH value of 11.0. The metal ions including K+, Ca2+, Na+, Mg2+, Mn2+, and Li+ strongly stimulated xanthan lyase activity but ions Zn2+ and Cu2+ were its inhibitor. Xanthan lyase should be a novel enzyme different from the other xanthan lyases ever reported. Fan Yang, Lan Yang, Xiaoyu Guo, Xue Wang, Lili Li, Zhicheng Liu, Wei Wang, and Xianzhen Li Copyright © 2014 Fan Yang et al. All rights reserved. Optimization of Processing Parameters for Extraction of Amylase Enzyme from Dragon (Hylocereus polyrhizus) Peel Using Response Surface Methodology Mon, 23 Jun 2014 12:23:29 +0000 The main goal of this study was to investigate the effect of extraction conditions on the enzymatic properties of thermoacidic amylase enzyme derived from dragon peel. The studied extraction variables were the buffer-to-sample (B/S) ratio (1 : 2 to 1 : 6, w/w), temperature (−18°C to 25°), mixing time (60 to 180 seconds), and the pH of the buffer (2.0 to 8.0). The results indicate that the enzyme extraction conditions exhibited the least significant () effect on temperature stability. Conversely, the extraction conditions had the most significant () effect on the specific activity and pH stability. The results also reveal that the main effect of the B/S ratio, followed by its interaction with the pH of the buffer, was significant () among most of the response variables studied. The optimum extraction condition caused the amylase to achieve high enzyme activity (648.4 U), specific activity (14.2 U/mg), temperature stability (88.4%), pH stability (85.2%), surfactant agent stability (87.2%), and storage stability (90.3%). Mehrnoush Amid, Mohd Yazid Abdul Manap, and Norkhanani Zohdi Copyright © 2014 Mehrnoush Amid et al. All rights reserved. N2O Emissions from an Apple Orchard in the Coastal Area of Bohai Bay, China Mon, 23 Jun 2014 07:58:55 +0000 Using static chambers and gas chromatography, nitrous oxide (N2O) fluxes from an apple orchard soil in the Bohai Bay region of China were measured from February 2010 to February 2011. In this study, two nitrogen (N) fertilizer treatments were designed—without (CK) or with (SN) synthetic N fertilizers (800 kg N ha−1). The annual cumulative N2O emissions from CK and SN were 34.6 ± 3.0 (mean ± standard error) and 44.3 ± 6.0 kg N2O–N ha−1, respectively. Such high emissions resulted from the intensive N fertilization in the experimental and previous years. The direct emission factor () of N2O induced by the applied synthetic N fertilizers was 1.2%. The is within the range of previous studies carried out in other croplands, which suggests that it is reasonable to estimate regional N2O emissions from apple orchards using the obtained in other croplands. In addition, significant positive correlations existed between N2O fluxes and soil temperatures or soil dissolved organic carbon contents. Baohua Xie, Junbao Yu, Xunhua Zheng, Fanzhu Qu, Yu Xu, and Haitao Lin Copyright © 2014 Baohua Xie et al. All rights reserved. Mine Land Reclamation and Eco-Reconstruction in Shanxi Province I: Mine Land Reclamation Model Sun, 22 Jun 2014 12:20:30 +0000 Coal resource is the main primary energy in our country, while Shanxi Province is the most important province in resource. Therefore Shanxi is an energy base for our country and has a great significance in energy strategy. However because of the heavy development of the coal resource, the ecological environment is worsening and the farmland is reducing continuously in Shanxi Province. How to resolve the contradiction between coal resource exploitation and environmental protection has become the imperative. Thus the concept of “green mining industry” is arousing more and more attention. In this assay, we will talk about the basic mode of land reclamation in mine area, the engineering study of mine land reclamation, the comprehensive model study of mine land reclamation, and the design and model of ecological agricultural reclamation in mining subsidence. Hao Bing-yuan and Kang Li-xun Copyright © 2014 Hao Bing-yuan and Kang Li-xun. All rights reserved. Micropropagation of Bioencapsulation and Ultrastructural Features of Sainfoin (Onobrychis viciifolia) Grown In Vivo and In Vitro Thu, 19 Jun 2014 12:46:56 +0000 To explore the potential of in vitro rapid regeneration, three varieties (Golpaygan-181, Orumieh-1763, and Gorgan-1601) of sainfoin (Onobrychis viciifolia Scop. syn. Onobrychis sativa L.) were evaluated. For the first time, an encapsulation protocol was established from somatic embryogenic callus in torpedo and cotyledonary stages to create artificial seeds. Callus derived from different concentrations of Kinetin (0–2.0 mg L−1) and Indole-3-acetic acid (0–2.0 mg L−1) was coated with sodium alginate and subsequently cultured either in Murashige and Skoog (MS) medium or in soil substrate. Adventitious shoots from synthetic beads developed into rooting in full and half strength MS medium supplemented with various concentrations of auxin and cytokinin. Prolonged water conservation of black and red soils (1 : 1) had the highest rate of survival plantlets in the acclimatization process. Diverse resistance techniques in Onobrychis viciifolia were evaluated when the plants were subjected to water deficiency. Higher frequency of epicuticular waxes was observed in in vivo leaves compared to in vitro leaves. Jagged trichomes nonsecreting glands covered by spines were only observed in the lower leaf side. Ultimately, stomata indices were 0.127 (abaxial), 0.188 (adaxial) in in vivo and 0.121 (abaxial), 0.201 (adaxial) in in vitro leaves. Sadegh Mohajer, Rosna Mat Taha, Minoo Mohajer, and Arash Khorasani Esmaeili Copyright © 2014 Sadegh Mohajer et al. All rights reserved. Regulating N Application for Rice Yield and Sustainable Eco-Agro Development in the Upper Reaches of Yellow River Basin, China Tue, 17 Jun 2014 11:02:17 +0000 High N fertilizer and flooding irrigation applied to rice on anthropogenic-alluvial soil often result in N leaching and low recovery of applied fertilizer N from the rice fields in Ningxia irrigation region in the upper reaches of the Yellow River, which threatens ecological environment, food security, and sustainable agricultural development. This paper reported the regulating N application for rice yield and sustainable Eco-Agro development in the upper reaches of Yellow River basin. The results showed that reducing and postponing N application could maintain crop yields while substantially reducing N leaching losses to the environment and improving the nitrogen use efficiency. Considering the high food production, the minimum environmental threat, and the low labor input, we suggested that regulating N application is an important measure to help sustainable agricultural development in this region. Aiping Zhang, Ruliang Liu, Ji Gao, Shiqi Yang, and Zhe Chen Copyright © 2014 Aiping Zhang et al. All rights reserved. Biodegradation of Palm Kernel Cake by Cellulolytic and Hemicellulolytic Bacterial Cultures through Solid State Fermentation Thu, 12 Jun 2014 06:27:28 +0000 Four cellulolytic and hemicellulolytic bacterial cultures were purchased from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Culture (DSMZ) and the American Type Culture Collection (ATCC). Two experiments were conducted; the objective of the first experiment was to determine the optimum time period required for solid state fermentation (SSF) of palm kernel cake (PKC), whereas the objective of the second experiment was to investigate the effect of combinations of these cellulolytic and hemicellulolytic bacteria on the nutritive quality of the PKC. In the first experiment, the SSF was lasted for 12 days with inoculum size of 10% (v/w) on different PKC to moisture ratios. In the second experiment, fifteen combinations were created among the four microbes with one untreated PKC as a control. The SSF lasted for 9 days, and the samples were autoclaved, dried, and analyzed for proximate analysis. Results showed that bacterial cultures produced high enzymes activities at the 4th day of SSF, whereas their abilities to produce enzymes tended to be decreased to reach zero at the 8th day of SSF. Findings in the second experiment showed that hemicellulose and cellulose was significantly decreased, whereas the amount of reducing sugars were significantly increased in the fermented PKC (FPKC) compared with untreated PKC. Mohamed Idris Alshelmani, Teck Chwen Loh, Hooi Ling Foo, Wei Hong Lau, and Awis Qurni Sazili Copyright © 2014 Mohamed Idris Alshelmani et al. All rights reserved. Functional Trait Trade-Offs for the Tropical Montane Rain Forest Species Responding to Light from Simulating Experiments Wed, 11 Jun 2014 06:51:03 +0000 Differences among tropical tree species in survival and growth to light play a key role in plant competition and community composition. Two canopy species with contrasting functional traits dominating early and late successional stages, respectively, in a tropical montane rain forest of Hainan Island, China, were selected in a pot experiment under 4 levels of light intensity (full, 50%, 30%, and 10%) in order to explore the adaptive strategies of tropical trees to light conditions. Under each light intensity level, the pioneer species, Endospermum chinense (Euphorbiaceae), had higher relative growth rate (RGR), stem mass ratio (SMR), specific leaf area (SLA), and morphological plasticity while the shade tolerant climax species, Parakmeria lotungensis (Magnoliaceae), had higher root mass ratio (RMR) and leaf mass ratio (LMR). RGR of both species was positively related to SMR and SLA under each light level but was negatively correlated with RMR under lower light (30% and 10% full light). The climax species increased its survival by a conservative resource use strategy through increasing leaf defense and root biomass investment at the expense of growth rate in low light. In contrast, the pioneer increased its growth by an exploitative resource use strategy through increasing leaf photosynthetic capacity and stem biomass investment at the expense of survival under low light. There was a trade-off between growth and survival for species under different light conditions. Our study suggests that tree species in the tropical rainforest adopt different strategies in stands of different successional stages. Species in the earlier successional stages have functional traits more advantageous to grow faster in the high light conditions, whereas species in the late successional stages have traits more favorable to survive in the low light conditions. Peili Mao, Runguo Zang, Hongbo Shao, and Junbao Yu Copyright © 2014 Peili Mao et al. All rights reserved. Differential Effects of Lichens versus Liverworts Epiphylls on Host Leaf Traits in the Tropical Montane Rainforest, Hainan Island, China Wed, 04 Jun 2014 08:33:22 +0000 Epiphylls widely colonize vascular leaves in moist tropical forests. Understanding the effects of epiphylls on leaf traits of host plants is critical for understanding ecological function of epiphylls. A study was conducted in a rain forest to investigate leaf traits of the host plants Photinia prunifolia colonized with epiphyllous liverworts and foliicolous lichens as well as those of uncolonized leaves. Our results found that the colonization of lichens significantly decreased leaf water content (LWC), chlorophyll (Chl) a and a + b content, and Chl a/b of P. prunifolia but increased Chl b content, while that of liverworts did not affect them as a whole. The variations of net photosynthetic rates among host leaves colonized with different coverage of lichens before or after removal treatment (a treatment to remove epiphylls from leaf surface) were greater than that colonized with liverworts. The full cover of lichens induced an increase of light compensation point (LCP) by 21% and a decrease of light saturation point (LSP) by 54% for their host leaves, whereas that of liverworts displayed contrary effects. Compared with the colonization of liverworts, lichens exhibited more negative effects on the leaf traits of P. prunifolia in different stages of colonization. The results suggest that the responses of host leaf traits to epiphylls are affected by the epiphyllous groups and coverage, which are also crucial factors in assessing ecofunctions of epiphylls in tropical forests. Lingyan Zhou, Fude Liu, Wenjie Yang, Hong Liu, Hongbo Shao, Zhongsheng Wang, and Shuqing An Copyright © 2014 Lingyan Zhou et al. All rights reserved. Potential of Pleurotus ostreatus Mycelium for Selenium Absorption Wed, 04 Jun 2014 07:37:06 +0000 The aim of this study was to evaluate the effect of high selenium (Se) concentrations on morphophysiological and ultrastructural properties of Pleurotus ostreatus. Mycelium growth was good in media enriched with 5.0, 10.0, and 20.0 mg L−1 of Se, concentration of 500.0 mg L−1 strongly inhibited growth, and 1000.0 mg L−1 was the minimum inhibitory concentration. Contrary to thin-walled, hyaline, branched, and anastomized hyphae with clamp-connections in the control, at Se concentrations of 100.0 and 500.0 mg L−1, they were noticeably short, frequently septed and branched, with a more intensive extracellular matrix, and without clamp-connections. At high Se concentrations, hyphae with intact membrane, without cellular contents, with a high level of vacuolization, and with numerous proteinaceous bodies were observed. Biomass yield ranged between 11.8 g L−1, in the control, and 6.8 g L−1, at an Se concentration of 100.0 mg L−1, while no production was detected at a concentration of 500.0 mg L−1. Se content in the mycelia reached a peak (938.9 μg g−1) after cultivation in the medium enriched with Se at the concentration of 20.0 mg L−1, while the highest absorption level (53.25%) was found in the medium enriched with 5.0 mg L−1 Se. Ivan Milovanović, Ilija Brčeski, Mirjana Stajić, Aleksandra Korać, Jelena Vukojević, and Aleksandar Knežević Copyright © 2014 Ivan Milovanović et al. All rights reserved. The Static Breaking Technique for Sustainable and Eco-Environmental Coal Mining Tue, 03 Jun 2014 11:15:03 +0000 The initiating explosive devices are prohibited in rock breaking near the goaf of the highly gassy mine. It is effective and applicable to cracking the hard roof with static cracking agent. By testing the static cracking of cubic limestone (size:  mm) with true triaxial rock mechanics testing machine under the effect of bidirectional stress and by monitoring the evolution process of the cracks generated during the acoustic emission experiment of static cracking, we conclude the following: the experiment results of the acoustic emission show that the cracks start from the lower part of the hole wall until they spread all over the sample. The crack growth rate follows a trend of “from rapidness to slowness.” The expansion time is different for the two bunches of cracks. The growth rates can be divided into the rapid increasing period and the rapid declining period, of which the growth rate in declining period is less than that in the increasing period. Also, the growth rate along the vertical direction is greater than that of the horizontal direction. Then the extended model for the static cracking is built according to the theories of elastic mechanics and fracture mechanics. Thus the relation formula between the applied forces of cracks and crack expansion radius is obtained. By comparison with the test results, the model proves to be applicable. In accordance with the actual geological situation of Yangquan No. 3 Mine, the basic parameters of manpower manipulated caving breaking with static crushing are settled, which reaps bumper industrial effects. Hao Bing-yuan, Huang Hui, Feng Zi-jun, and Wang Kai Copyright © 2014 Hao Bing-yuan et al. All rights reserved. Efficient Reprogramming of Mouse Fibroblasts to Neuronal Cells including Dopaminergic Neurons Sun, 01 Jun 2014 11:22:09 +0000 Somatic cells were directly converted to functional neurons through the use of a combination of transcription factors, including Ascl1, Brn2, and Myt1l. However, a major limitation is the lack of a reliable source of cell-replacement therapy for neurological diseases. Here, we show that a combination of the transcription factors Ascl1 and Nurr1 (AN) and neurotrophic factors including SHH and FGF8b directly reprogrammed embryonic mouse fibroblasts to induced neuronal (iN) cells: pan-neuronal cells and dopaminergic (DA) neurons under our systematic cell culture conditions. Reprogrammed cells showed the morphological properties of neuronal cells. Additionally, cells were analyzed using various markers, including Tuj1 and Map2 for neuronal cells and Lmx1a, Th, Aadc and Vmat2 for DA neurons in our immunostaining and reverse transcription (RT)-PCR experiments. We found that a combination of transcription factors and neurotrophic factors could directly reprogram fibroblasts to neuronal cells including DA neurons. Various types of reprogrammed cells are promising cell sources for cell-based therapy of neurological disorders like Parkinson’s disease and spinal cord injury. Seung-ick Oh, Hang-soo Park, Insik Hwang, Han-kyul Park, Kyung-Ah Choi, Hyesun Jeong, Suhng Wook Kim, and Sunghoi Hong Copyright © 2014 Seung-ick Oh et al. All rights reserved. Interactive Effects of Moss-Dominated Crusts and Artemisia ordosica on Wind Erosion and Soil Moisture in Mu Us Sandland, China Sun, 01 Jun 2014 11:17:40 +0000 To better understand the effects of biological soil crusts (BSCs) on soil moisture and wind erosion and study the necessity and feasibility of disturbance of BSCs in the Mu Us sandland, the effects of four treatments, including moss-dominated crusts alone, Artemisia ordosica alone, bare sand, and Artemisia ordosica combined with moss-dominated crusts, on rainwater infiltration, soil moisture, and annual wind erosion were observed. The major results are as follows. (1) The development of moss-dominated crusts exacerbated soil moisture consumption and had negative effects on soil moisture in the Mu Us sandland. (2) Moss-dominated crusts significantly increased soil resistance to wind erosion, and when combined with Artemisia ordosica, this effect became more significant. The contribution of moss-dominated crusts under Artemisia ordosica was significantly lower than that of moss-dominated crusts alone in sites where vegetative coverage > 50%. (3) Finally, an appropriate disturbance of moss-dominated crusts in the rainy season in sites with high vegetative coverage improved soil water environment and vegetation succession, but disturbance in sites with little or no vegetative cover should be prohibited to avoid the exacerbation of wind erosion. Yongsheng Yang, Chongfeng Bu, Xingmin Mu, Hongbo Shao, and Kankan Zhang Copyright © 2014 Yongsheng Yang et al. All rights reserved. Hydrogen Peroxide Is Involved in Salicylic Acid-Elicited Rosmarinic Acid Production in Salvia miltiorrhiza Cell Cultures Thu, 29 May 2014 14:34:47 +0000 Salicylic acid (SA) is an elicitor to induce the biosynthesis of secondary metabolites in plant cells. Hydrogen peroxide (H2O2) plays an important role as a key signaling molecule in response to various stimuli and is involved in the accumulation of secondary metabolites. However, the relationship between them is unclear and their synergetic functions on accumulation of secondary metabolites are unknown. In this paper, the roles of SA and H2O2 in rosmarinic acid (RA) production in Salvia miltiorrhiza cell cultures were investigated. The results showed that SA significantly enhanced H2O2 production, phenylalanine ammonia-lyase (PAL) activity, and RA accumulation. Exogenous H2O2 could also promote PAL activity and enhance RA production. If H2O2 production was inhibited by NADPH oxidase inhibitor (IMD) or scavenged by quencher (DMTU), RA accumulation would be blocked. These results indicated that H2O2 is secondary messenger for signal transduction, which can be induced by SA, significantly and promotes RA accumulation. Wenfang Hao, Hongbo Guo, Jingyi Zhang, Gege Hu, Yaqin Yao, and Juane Dong Copyright © 2014 Wenfang Hao et al. All rights reserved. A Meta-Analysis of the Bacterial and Archaeal Diversity Observed in Wetland Soils Wed, 28 May 2014 09:00:53 +0000 This study examined the bacterial and archaeal diversity from a worldwide range of wetlands soils and sediments using a meta-analysis approach. All available 16S rRNA gene sequences recovered from wetlands in public databases were retrieved. In November 2012, a total of 12677 bacterial and 1747 archaeal sequences were collected in GenBank. All the bacterial sequences were assigned into 6383 operational taxonomic units (OTUs 0.03), representing 31 known bacterial phyla, predominant with Proteobacteria (2791 OTUs), Bacteroidetes (868 OTUs), Acidobacteria (731 OTUs), Firmicutes (540 OTUs), and Actinobacteria (418 OTUs). The genus Flavobacterium (11.6% of bacterial sequences) was the dominate bacteria in wetlands, followed by Gp1, Nitrosospira, and Nitrosomonas. Archaeal sequences were assigned to 521 OTUs from phyla Euryarchaeota and Crenarchaeota. The dominating archaeal genera were Fervidicoccus and Methanosaeta. Rarefaction analysis indicated that approximately 40% of bacterial and 83% of archaeal diversity in wetland soils and sediments have been presented. Our results should be significant for well-understanding the microbial diversity involved in worldwide wetlands. Xiaofei Lv, Junbao Yu, Yuqin Fu, Bin Ma, Fanzhu Qu, Kai Ning, and Huifeng Wu Copyright © 2014 Xiaofei Lv et al. All rights reserved. Essential Oil Composition and Antimicrobial Activities of Two Closely Related Species, Alpinia mutica Roxb. and Alpinia latilabris Ridl., from Peninsular Malaysia Tue, 27 May 2014 12:27:49 +0000 The essential oils obtained by hydrodistillation of the unripe and ripe fruits of Alpinia mutica Roxb. and Alpinia latilabris Ridl. were analysed by capillary GC and GC-MS. The oils were principally monoterpenic in nature. The unripe and ripe fruit oils of A. mutica were characterized by camphor (21.0% and 15.8%), camphene (16.6% and 10.2%), β-pinene (8.6% and 13.5%), and trans,trans-farnesol (8.0% and 11.2%), respectively. The oils of the unripe and ripe fruits were moderately active against Staphylococcus aureus, Bacillus subtilis, Trichophyton mentagrophytes, and Trichophyton rubrum. 1,8-Cineole (34.2% and 35.9%) and β-pinene (20.2% and 19.0%) were the two most abundant components in the unripe and ripe fruit oils of A. latilabris. The oil of the unripe fruits elicits moderate activity against Staphylococcus aureus and Trichophyton mentagrophytes while Candida glabrata was moderately sensitive to the oil of the ripe fruits. Halijah Ibrahim, Yasodha Sivasothy, Devi Rosmy Syamsir, Noor Hasima Nagoor, Natasha Jamil, and Khalijah Awang Copyright © 2014 Halijah Ibrahim et al. All rights reserved. Reference Gene Selection for Quantitative Real-Time RT-PCR Normalization in Iris. lactea var. chinensis Roots under Cadmium, Lead, and Salt Stress Conditions Mon, 26 May 2014 11:15:41 +0000 Quantitative real time PCR (RT-qPCR) has emerged as an accurate and sensitive method to measure the gene expression. However, obtaining reliable result depends on the selection of reference genes which normalize differences among samples. In this study, we assessed the expression stability of seven reference genes, namely, ubiquitin-protein ligase UBC9 (UBC), tubulin alpha-5 (TUBLIN), eukaryotic translation initiation factor (EIF-5A), translation elongation factor EF1A (EF1α), translation elongation factor EF1B (EF1b), actin11 (ACTIN), and histone H3 (HIS), in Iris. lactea var. chinensis (I. lactea var. chinensis) root when the plants were subjected to cadmium (Cd), lead (Pb), and salt stress conditions. All seven reference genes showed a relatively wide range of threshold cycles () values in different samples. GeNorm and NormFinder algorithms were used to assess the suitable reference genes. The results from the two software units showed that EIF-5A and UBC were the most stable reference genes across all of the tested samples, while TUBLIN was unsuitable as internal controls. I. lactea var. chinensis is tolerant to Cd, Pb, and salt. Our results will benefit future research on gene expression in response to the three abiotic stresses. Chun-Sun Gu, Liang-qin Liu, Chen Xu, Yan-hai Zhao, Xu-dong Zhu, and Su-Zhen Huang Copyright © 2014 Chun-Sun Gu et al. All rights reserved. Effects of Silicon on Photosynthetic Characteristics of Maize (Zea mays L.) on Alluvial Soil Mon, 26 May 2014 07:22:07 +0000 The objectives of the study were to determine the effects of silicon on photosynthetic characteristics of maize on alluvial soil, including total chlorophyll contents, photosynthetic rate , stomatal conductance , transpiration rate (), and intercellular CO2 concentration using the method of field experiment, in which there were five levels (0, 45, 90, 150, and 225 kg·ha−1) of silicon supplying. The results showed that certain doses of silicon fertilizers can be used successfully in increasing the values of total chlorophyll contents, , and and decreasing the values of and of maize leaves, which meant that photosynthetic efficiency of maize was significantly increased in different growth stages by proper doses of Si application on alluvial soil, and the optimal dose of Si application was 150 kg·ha−1. Our results indicated that silicon in proper amounts can be beneficial in increasing the photosynthetic ability of maize, which would be helpful for the grain yield and growth of maize. Zhiming Xie, Fengbin Song, Hongwen Xu, Hongbo Shao, and Ri Song Copyright © 2014 Zhiming Xie et al. All rights reserved. Comparative Ecophysiological Study of Salt Stress for Wild and Cultivated Soybean Species from the Yellow River Delta, China Sun, 25 May 2014 11:56:52 +0000 Osmotic and ionic stresses were the primary and instant damage produced by salt stress. They can also bring about other secondary stresses. Soybean is an important economic crop and the wild soybean aroused increasing attention for its excellent performance in salt resistance. For this reason, we compared the different performances of Glycine max L. (ZH13) and Glycine soja L. (BB52) in both young and mature seedlings, hoping to clarify the specific reasons. Our research revealed that, compared to the cultivated soybean, the wild soybean was able to maintain higher water potential and relative water content (RWC), accumulate more amount of proline and glycine betaine, reduce the contents of Na+ and Cl− by faster efflux, and cut down the efflux of the K+ as well as keep higher K+/Na+ ratio. And what is more is that, almost all the excel behaviors became particularly obvious under higher NaCl concentration (300 mM). Therefore, according to all the detections and comparisons, we concluded that the wild soybean had different tolerance mechanisms and better salt resistance. It should be used as eminent germplasm resource to enhance the resistant ability of cultivated soybean or even other crops. Gang Wu, Zhengda Zhou, Peng Chen, Xiaoli Tang, Hongbo Shao, and Hongyan Wang Copyright © 2014 Gang Wu et al. All rights reserved. Distribution Characteristic of Soil Organic Carbon Fraction in Different Types of Wetland in Hongze Lake of China Thu, 22 May 2014 11:33:17 +0000 Soil organic carbon fractions included microbial biomass carbon (MBC), dissolved organic carbon (DOC), and labile organic carbon (LOC), which was investigated over a 0–20 cm depth profile in three types of wetland in Hongze Lake of China. Their ecoenvironmental effect and the relationships with soil organic carbon (SOC) were analyzed in present experiment. The results showed that both active and SOC contents were in order reduced by estuarine wetland, flood plain, and out-of-lake wetland. Pearson correlative analysis indicated that MBC and DOC were positively related to SOC. The lowest ratios of MBC and DOC to SOC in the estuarine wetland suggested that the turnover rate of microbial active carbon pool was fairly low in this kind of wetland. Our results showed that estuarine wetland had a strong carbon sink function, which played important role in reducing greenhouse gas emissions; besides, changes of water condition might affect the accumulation and decomposition of organic carbon in the wetland soils. Yan Lu and Hongwen Xu Copyright © 2014 Yan Lu and Hongwen Xu. All rights reserved. An Effective Protocol for Micropropagation of Edible Bamboo Species (Bambusa tulda and Melocanna baccifera) through Nodal Culture Wed, 21 May 2014 00:00:00 +0000 High demand for edible bamboo shoots of Bambusa tulda and Melocanna baccifera in many Asian ethnic groups has led to the need for developing intensive bamboo farming. To achieve this, in vitro regeneration of bamboo plantlets is needed due to the long and irregular bamboo flowering cycle and scarcity of bamboo seeds. An effective protocol for plantlets regeneration in B. tulda and M. baccifera from nodal explants following validation of the species using the sequence of trnL-F intergenic spacer region is described. Effective axillary bud breaking was achieved at 3 mg/L of 6-benzylaminopurine (BAP) in MS medium. Importantly, combining 2 mg/L of kinetin (Kn) with 3 mg/L of BAP produced a synergistic effect for shoot multiplication in B. tulda and M. baccifera. Under optimized conditions in half-strength MS medium supplemented with 3 mg/L of indole-3-butyric acid (IBA), 10 mg/L of coumarin, and 3% sucrose, profuse production of dark-brown rhizome in B. tulda and abundant rooting (81.67%, , ) for M. baccifera within 30 days were achieved. The established protocol and the validation of the reported species at the molecular level will be of help to stakeholders in edible bamboo trade to conserve gene-pool and increase productivity. Sayanika Devi Waikhom and Bengyella Louis Copyright © 2014 Sayanika Devi Waikhom and Bengyella Louis. All rights reserved. Soil Phosphorus Forms and Profile Distributions in the Tidal River Network Region in the Yellow River Delta Estuary Tue, 20 May 2014 00:00:00 +0000 Modified Hedley fraction method was used to study the forms and profile distribution in the tidal river network region subjected to rapid deposition and hydrologic disturbance in the Yellow River Delta (YRD) estuary, eastern China. The results showed that the total P () ranged from 612.1 to 657.8 mg kg−1. Dilute HCl extractable inorganic P () was the predominant form in all profiles, both as absolute values and as a percentage of total extracted . The NaOH extractable organic P () was the predominant form of total extracted , while Bicarb- and C.HCl- were the lowest fractions of total extracted and in all the P forms. The Resin-P concentrations were high in the top soil layer and decreased with depth. The Pearson correlation matrix indicated that Resin-P, Bicarb-, NaOH-, and C.HCl- were strongly positively correlated with salinity, TOC, Ca, Al, and Fe but negatively correlated with pH. The significant correlation of any studied form of organic P (Bicarb-, NaOH-, and C.HCl-) with geochemical properties were not observed in the study. Duncan multiple-range test indicated that the P forms and distribution heterogeneity in the profiles could be attributed to the influences of vegetation cover and hydrologic disturbance. Junbao Yu, Fanzhu Qu, Huifeng Wu, Ling Meng, Siyao Du, and Baohua Xie Copyright © 2014 Junbao Yu et al. All rights reserved. Soil Characteristic Comparison of Fenced and Grazed Riparian Floodplain Wetlands in the Typical Steppe Region of the Inner Mongolian Plateau, China Mon, 19 May 2014 00:00:00 +0000 In recent decades, degradation of ecosystem in the steppe region of the Inner Mongolia Plateau, especially in riparian floodplain wetlands, has become a significant ecological crisis. Not uncommonly, with the increasing of livestock in the Inner Mongolian steppe region, a riparian floodplain wetland is becoming a hotspot area of grazing for local herdsmen. Hence, it is essential to understand degradation mechanisms of riparian floodplain wetland ecosystems caused by extensive grazing. In this study, the spatial distribution of soil compaction, salinity, total nitrogen, total phosphorus, organic carbon, and microbial biomass C and N were investigated. The results showed that grazing led to an increase in soil compaction and soil surface salinity, which significantly lowered levels of total N, P, and TOC in the soil surface. Grazing decreased soil microbial biomass C and N concentration in the lower riparian floodplain wetland, whereas it significantly increased soil microbial biomass C and N concentration in the higher riparian floodplain wetland. Elevation differences in the riparian floodplain wetland increased spatial heterogeneity in the soil and thus resulted in different influence of grazing on wetland soils and ecosystem. Therefore, elevation differences and grazing intensity were the main factors controlling soil characteristics in the riparian floodplain wetland of this region. Lixin Wang, Huamin Liu, Yuhong Liu, Jianwei Li, Hongbo Shao, Wei Wang, and Cunzhu Liang Copyright © 2014 Lixin Wang et al. All rights reserved. Effects of Age and Stand Density of Mother Trees on Early Pinus thunbergii Seedling Establishment in the Coastal Zone, China Thu, 15 May 2014 12:33:31 +0000 Effects of age and stand density of mother tree on seed germination, seedling biomass allocation, and seedling growth of Pinus thunbergii were studied. The results showed that age of mother tree did not have significant influences on seed germination, but it was significant on seedling biomass allocation and growth. Seedlings from the minimum and maximum age of mother tree had higher leaf mass ratio and lower root mass ratio than from the middle age of mother tree. Moreover, they also had higher relative height growth rate and slenderness, which were related to their biomass allocation. Stand density of mother tree mainly demonstrated significant effects on seed germination and seedling growth. Seed from higher stand density of mother tree did not decrease germination rate, but had higher mean germination time, indicating that it delayed germination process. Seedlings of higher stand density of mother tree showed higher relative height growth rate and slenderness. These traits of offspring from higher stand density of mother tree were similar to its mother, indicating significant environmental maternal effects. So, mother tree identity of maternal age and environments had important effects on natural regeneration of the coastal P. thunbergii forest. Peili Mao, Guangxuan Han, Guangmei Wang, Junbao Yu, and Hongbo Shao Copyright © 2014 Peili Mao et al. All rights reserved. Variation of Runoff and Precipitation in the Hekou-Longmen Region of the Yellow River Based on Elasticity Analysis Mon, 12 May 2014 12:16:21 +0000 Precipitation is very important to the formation of runoff, and studying of runoff variation and its response to precipitation has practical significance to sustainable utilization of water resources. The study used Mann-Kendall test, anomaly accumulation method, and precipitation elasticity of runoff method to analyze the changes in the relation of precipitation and runoff and the contribution of precipitation to runoff change in the Hekou-Longmen region (from 1957 to 2010), Huangfuchuan watershed (from 1954 to 2010), and Yanhe watershed (from 1952 to 2010) in the middle reaches of the Yellow River. The results showed that runoff appeared a significant decreasing trend while it was not significant in precipitation in all study areas. In particular, the reductions of average annual runoff in the Hekou-Longmen region, Huangfuchuan watershed, and Yanhe watershed were 72.7%, 87.5%, and 32.2%, respectively, during 2000–2010 compared to the 1950s. There existed two abrupt change points of the runoff in the Hekou-Longmen region and Huangfuchuan watershed, which were detected in 1979 and 1998. But in the Yanhe watershed only one abrupt change point was found in 1996. The precipitation elasticities of runoff were 1.11, 1.09, and 1.26, respectively, and the contributions of precipitation on runoff reduction were 26.4%, 17.9%, and 31.6%, respectively, in the Hekou-Longmen region, Huangfuchuan watershed, and Yanhe watershed. Erhui Li, Xingmin Mu, Guangju Zhao, Peng Gao, and Hongbo Shao Copyright © 2014 Erhui Li et al. All rights reserved. Effects of NAA and BAP, Double-Layered Media, and Light Distance on In Vitro Regeneration of Nelumbo nucifera Gaertn. (Lotus), an Aquatic Edible Plant Wed, 07 May 2014 13:15:30 +0000 In vitro direct regeneration of Nelumbo nucifera Gaertn. was successfully achieved from immature explants (yellow plumule) cultured on a solid MS media supplemented with combinations of 0.5 mg/L BAP and 1.5 mg/L NAA which resulted in 16.00 ± 0.30 number of shoots per explant and exhibited a new characteristic of layered multiple shoots, while normal roots formed on the solid MS basal media. The double-layered media gave the highest number of shoots per explant with a ratio of 2 : 1 (liquid to solid) with a mean number of 16.67 ± 0.23 shoots per explant with the formation of primary and secondary roots from immature explants. In the study involving light distance, the tallest shoot (16.67 ± 0.23 mm) obtained from the immature explants was at a light distance of 200 mm from the source of inflorescent light (1000 lux). The plantlets were successfully acclimatized in clay loam soil after 8 months being maintained under in vitro conditions. Noraini Mahmad, Rosna Mat Taha, Rashidi Othman, Azani Saleh, Nor Azlina Hasbullah, and Hashimah Elias Copyright © 2014 Noraini Mahmad et al. All rights reserved. Effects of Different Vegetation Zones on CH4 and N2O Emissions in Coastal Wetlands: A Model Case Study Tue, 29 Apr 2014 12:07:59 +0000 The coastal wetland ecosystems are important in the global carbon and nitrogen cycle and global climate change. For higher fragility of coastal wetlands induced by human activities, the roles of coastal wetland ecosystems in CH4 and N2O emissions are becoming more important. This study used a DNDC model to simulate current and future CH4 and N2O emissions of coastal wetlands in four sites along the latitude in China. The simulation results showed that different vegetation zones, including bare beach, Spartina beach, and Phragmites beach, produced different emissions of CH4 and N2O in the same latitude region. Correlation analysis indicated that vegetation types, water level, temperature, and soil organic carbon content are the main factors affecting emissions of CH4 and N2O in coastal wetlands. Yuhong Liu, Lixin Wang, Shumei Bao, Huamin Liu, Junbao Yu, Yu Wang, Hongbo Shao, Yan Ouyang, and Shuqing An Copyright © 2014 Yuhong Liu et al. All rights reserved. Effects of Thinning Intensities on Soil Infiltration and Water Storage Capacity in a Chinese Pine-Oak Mixed Forest Wed, 23 Apr 2014 07:18:12 +0000 Thinning is a crucial practice in the forest ecosystem management. The soil infiltration rate and water storage capacity of pine-oak mixed forest under three different thinning intensity treatments (15%, 30%, and 60%) were studied in Qinling Mountains of China. The thinning operations had a significant influence on soil infiltration rate and water storage capacity. The soil infiltration rate and water storage capacity in different thinning treatments followed the order of control (nonthinning): <60%, <15%, and <30%. It demonstrated that thinning operation with 30% intensity can substantially improve soil infiltration rate and water storage capacity of pine-oak mixed forest in Qinling Mountains. The soil initial infiltration rate, stable infiltration rate, and average infiltration rate in thinning 30% treatment were significantly increased by 21.1%, 104.6%, and 60.9%, compared with the control. The soil maximal water storage capacity and noncapillary water storage capacity in thinning 30% treatment were significantly improved by 20.1% and 34.3% in contrast to the control. The soil infiltration rate and water storage capacity were significantly higher in the surface layer (0~20 cm) than in the deep layers (20~40 cm and 40~60 cm). We found that the soil property was closely related to soil infiltration rate and water storage capacity. Lili Chen, Zhiyou Yuan, Hongbo Shao, Dexiang Wang, and Xingmin Mu Copyright © 2014 Lili Chen et al. All rights reserved. Multivariate-Statistical Assessment of Heavy Metals for Agricultural Soils in Northern China Wed, 16 Apr 2014 12:56:41 +0000 The study evaluated eight heavy metals content and soil pollution from agricultural soils in northern China. Multivariate and geostatistical analysis approaches were used to determine the anthropogenic and natural contribution of soil heavy metal concentrations. Single pollution index and integrated pollution index could be used to evaluate soil heavy metal risk. The results show that the first factor explains 27.3% of the eight soil heavy metals with strong positive loadings on Cu, Zn, and Cd, which indicates that Cu, Zn, and Cd are associated with and controlled by anthropic activities. The average value of heavy metal is lower than the second grade standard values of soil environmental quality standards in China. Single pollution index is lower than 1, and the Nemerow integrated pollution index is 0.305, which means that study area has not been polluted. The semivariograms of soil heavy metal single pollution index fitted spherical and exponential models. The variable ratio of single pollution index showed moderately spatial dependence. Heavy metal contents showed relative safety in the study area. Pingguo Yang, Miao Yang, Renzhao Mao, and Hongbo Shao Copyright © 2014 Pingguo Yang et al. All rights reserved. Tissue Fractions of Cadmium in Two Hyperaccumulating Jerusalem Artichoke Genotypes Mon, 14 Apr 2014 12:45:00 +0000 In order to investigate the mechanisms in two Jerusalem artichoke (Helianthus tuberosus L.) genotypes that hyperaccumulate Cd, a sand-culture experiment was carried out to characterize fractionation of Cd in tissue of Cd-hyperaccumulating genotypes NY2 and NY5. The sequential extractants were: 80% v/v ethanol (), deionized water (), 1 M NaCl (), 2% v/v acetic acid (), and 0.6 M HCl (). After 20 days of treatments, NY5 had greater plant biomass and greater Cd accumulation in tissues than NY2. In both genotypes the fraction was the highest in roots and stems, whereas the and fractions were the highest in leaves. With an increase in Cd concentration in the culture solution, the content of every Cd fraction also increased. The and ratios in roots were lower in NY5 than in NY2, while the amount of other Cd forms was higher. It implied that, in high accumulator, namely, NY5, the complex of insoluble phosphate tends to be shaped more easily which was much better for Cd accumulation. Besides, translocation from plasma to vacuole after combination with protein may be one of the main mechanisms in Cd-accumulator Jerusalem artichoke genotypes. Xiaohua Long, Ni Ni, Zhaopu Liu, Zed Rengel, Xin Jiang, and Hongbo Shao Copyright © 2014 Xiaohua Long et al. All rights reserved. Effects of Urbanization Expansion on Landscape Pattern and Region Ecological Risk in Chinese Coastal City: A Case Study of Yantai City Thu, 10 Apr 2014 17:51:45 +0000 Applied with remote sensing, GIS, and mathematical statistics, the spatial-temporal evolution characteristics of urbanization expansion of Yantai city from 1974 to 2009 was studied. Based on landscape pattern metrics and ecological risk index, the landscape ecological risk from the landscape pattern dynamics was evaluated. The results showed that the area of urban land increased by 189.77 km2 with average expansion area of 5.42 km2 y−1 from 1974 to 2009. The urbanization intensity index during 2004–2009 was 3.92 times of that during 1974–1990. The land use types of urban land and farmland changed greatly. The changes of landscape pattern metrics for land use patterns indicated that the intensity of human activities had strengthened gradually in study period. The landscape ecological risk pattern of Yantai city shaped half-round rings along the coastline. The ecological risk index decreased with increase of the distance to the coastline. The ratio of high ecological risk to subhigh ecological risk zones in 2009 was 2.23 times of that in 1990. The significant linear relationship of urbanization intensity index and regional ecological risk indicated that the anthropological economic activities were decisive factors for sustainable development of costal ecological environment. Di Zhou, Ping Shi, Xiaoqing Wu, Jinwei Ma, and Junbao Yu Copyright © 2014 Di Zhou et al. All rights reserved. Agrobacterium-Mediated Transformation of the Recalcitrant Vanda Kasem's Delight Orchid with Higher Efficiency Tue, 08 Apr 2014 07:56:17 +0000 The presented study established Agrobacterium-mediated genetic transformation using protocorm-like bodies (PLBs) for the production of transgenic Vanda Kasem's Delight Tom Boykin (VKD) orchid. Several parameters such as PLB size, immersion period, level of wounding, Agrobacterium density, cocultivation period, and concentration of acetosyringone were tested and quantified using gusA gene expression to optimize the efficiency of Agrobacterium-mediated genetic transformation of VKD's PLBs. Based on the results, 3-4 mm PLBs wounded by scalpel and immersed for 30 minutes in Agrobacterium suspension of 0.8 unit at produced the highest GUS expression. Furthermore, cocultivating infected PLBs for 4 days in the dark on Vacin and Went cocultivation medium containing 200 𝜇M acetosyringone enhanced the GUS expression. PCR analysis of the putative transformants selected in the presence of 250 mg/L cefotaxime and 30 mg/L geneticin proved the presence of wheatwin1, wheatwin2, and nptII genes. Pavallekoodi Gnasekaran, Jessica Jeyanthi James Antony, Jasim Uddain, and Sreeramanan Subramaniam Copyright © 2014 Pavallekoodi Gnasekaran et al. All rights reserved. Optimization of Culture Conditions (Sucrose, pH, and Photoperiod) for In Vitro Regeneration and Early Detection of Somaclonal Variation in Ginger Lime (Citrus assamensis) Tue, 01 Apr 2014 09:32:27 +0000 Various explants (stem, leaf, and root) of Citrus assamensis were cultured on MS media supplemented with various combinations and concentrations (0.5–2.0 mgL−1) of NAA and BAP. Optimum shoot and root regeneration were obtained from stem cultures supplemented with 1.5 mgL−1 NAA and 2.0 mgL−1 BAP, respectively. Explant type affects the success of tissue culture of this species, whereby stem explants were observed to be the most responsive. Addition of 30 gL−1 sucrose and pH of 5.8 was most optimum for in vitro regeneration of this species. Photoperiod of 16 hours of light and 8 hours of darkness was most optimum for shoot regeneration, but photoperiod of 24 hours of darkness was beneficial for production of callus. The morphology (macro and micro) and anatomy of in vivo and in vitro/ex vitro Citrus assamensis were also observed to elucidate any irregularities (or somaclonal variation) that may arise due to tissue culture protocols. Several minor micromorphological and anatomical differences were observed, possibly due to stress of tissue culture, but in vitro plantlets are expected to revert back to normal phenotype following full adaptation to the natural environment. Jamilah Syafawati Yaacob, Noraini Mahmad, Rosna Mat Taha, Normadiha Mohamed, Anis Idayu Mad Yussof, and Azani Saleh Copyright © 2014 Jamilah Syafawati Yaacob et al. All rights reserved. Wet and Dry Atmospheric Depositions of Inorganic Nitrogen during Plant Growing Season in the Coastal Zone of Yellow River Delta Tue, 01 Apr 2014 09:07:46 +0000 The ecological problems caused by dry and wet deposition of atmospheric nitrogen have been widespread concern in the world. In this study, wet and dry atmospheric depositions were monitored in plant growing season in the coastal zone of the Yellow River Delta (YRD) using automatic sampling equipment. The results showed that and Na+ were the predominant anion and cation, respectively, in both wet and dry atmospheric depositions. The total atmospheric nitrogen deposition was ~2264.24 mg m−2, in which dry atmospheric nitrogen deposition was about 32.02%. The highest values of dry and wet atmospheric nitrogen deposition appeared in May and August, respectively. In the studied area, –N was the main nitrogen form in dry deposition, while the predominant nitrogen in wet atmospheric deposition was –N with ~56.51% of total wet atmospheric nitrogen deposition. The average monthly attribution rate of atmospheric deposition of –N and –N was ~31.38% and ~20.50% for the contents of –N and –N in 0–10 cm soil layer, respectively, suggested that the atmospheric nitrogen was one of main sources for soil nitrogen in coastal zone of the YRD. Junbao Yu, Kai Ning, Yunzhao Li, Siyao Du, Guangxuan Han, Qinghui Xing, Huifeng Wu, Guangmei Wang, and Yongjun Gao Copyright © 2014 Junbao Yu et al. All rights reserved. Comparison of Microtox and Xenoassay Light as a Near Real Time River Monitoring Assay for Heavy Metals Tue, 01 Apr 2014 07:58:12 +0000 Luminescence-based assays for toxicants such as Microtox, ToxAlert, and Biotox have been used extensively worldwide. However, the use of these assays in near real time conditions is limited due to nonoptimal assay temperature for the tropical climate. An isolate that exhibits a high luminescence activity in a broad range of temperatures was successfully isolated from the mackerel, Rastrelliger kanagurta. This isolate was tentatively identified as Photobacterium sp. strain MIE, based on partial 16S rDNA molecular phylogeny. Optimum conditions that support high bioluminescence activity occurred between 24 and 30°C, with pH 5.5 to 7.5, 10 to 20 g/L of sodium chloride, 30 to 50 g/L of tryptone, and 4 g/L of glycerol as the carbon source. Assessment of near real time capability of this bacterial system, Xenoassay light to monitor heavy metals from a contaminated river running through the Juru River Basin shows near real time capability with assaying time of less than 30 minutes per samples. Samples returned to the lab were tested with a standard Microtox assay using Vibrio fishceri. Similar results were obtained to Xenoassay light that show temporal variation of copper concentration. Thus, this strain is suitable for near real time river monitoring of toxicants especially in the tropics. M. I. E. Halmi, Hussain Jirangon, W. L. W. Johari, A. R. Abdul Rachman, M. Y. Shukor, and M. A. Syed Copyright © 2014 M. I. E. Halmi et al. All rights reserved. Ecological Effects of Roads on the Plant Diversity of Coastal Wetland in the Yellow River Delta Mon, 31 Mar 2014 16:59:00 +0000 The 26 sample sites in 7 study plots adjacent to asphalt road and earth road in coastal wetland in the Yellow River Delta were selected to quantify plant diversity using quadrat sampling method in plant bloom phase of July and August 2012. The indice of and Jaccard’s coefficient were applied to evaluate the species diversity. The results showed that the plant diversities and alien plants were high in the range of 0–20 m to the road verge. There were more exotics and halophytes in plots of asphalt roadside than that of earth roadside. However, proportion of halophytes in habitats of asphalt roadsides was lower than that of earth roadside. By comparing -diversity, there were more common species in the asphalt roadsides than that in the earth roadsides. The similarity of plant communities in studied plots of asphalt roadsides and earth roadsides increased with increasing the distance to road verge. The effect range of roads for plant diversity in study region was about 20 m to road verge. Our results indicate that the construction and maintenance of roads in wetland could increase the plant species diversities of communities and risk of alien species invasion. Yunzhao Li, Junbao Yu, Kai Ning, Siyao Du, Guangxuan Han, Fanzhu Qu, Guangmei Wang, Yuqin Fu, and Chao Zhan Copyright © 2014 Yunzhao Li et al. All rights reserved. Fluorescence Enhancement of Fluorescent Unnatural Streptavidin by Binding of a Biotin Analogue with Spacer Tail and Its Application to Biotin Sensing Thu, 20 Mar 2014 13:52:07 +0000 We designed a novel molecular biosensing system for the detection of biotin, an important vitamin by the combination of fluorescent unnatural streptavidin with a commercialized biotin-(AC5)2-hydrazide. A fluorescent unnatural amino acid, BODIPY-FL-aminophenylalanine (BFLAF), was position-specifically incorporated into Trp120 of streptavidin by four-base codon method. Fluorescence of the Trp120BFLAF mutant streptavidin was enhanced by the addition of biotin-(AC5)2-hydrazide with the concentration dependent, whereas fluorescence enhancement was not observed at all by the addition of natural biotin. It was considered that the spacer tail of biotin-(AC5)2-hydrazide may disturb the fluorescence quenching of the Trp120BFLAF by Trp79 and Trp108 of the neighbor subunit. Therefore, biotin sensing was carried out by the competitive binding reaction of biotin-(AC5)2-hydrazide and natural biotin to the fluorescent mutant streptavidin. The fluorescence intensity decreased by increasing free biotin concentration. The result suggested that molecular biosensor for small ligand could be successfully designed by the pair of fluorescent mutant binding protein and ligand analogue. Xianwei Zhu and Hiroaki Shinohara Copyright © 2014 Xianwei Zhu and Hiroaki Shinohara. All rights reserved. Overexpression of the Synthetic Chimeric Native-T-phylloplanin-GFP Genes Optimized for Monocot and Dicot Plants Renders Enhanced Resistance to Blue Mold Disease in Tobacco (N. tabacum L.) Thu, 20 Mar 2014 09:38:41 +0000 To enhance the natural plant resistance and to evaluate the antimicrobial properties of phylloplanin against blue mold, we have expressed a synthetic chimeric native-phylloplanin-GFP protein fusion in transgenic Nicotiana tabacum cv. KY14, a cultivar that is highly susceptible to infection by Peronospora tabacina. The coding sequence of the tobacco phylloplanin gene along with its native signal peptide was fused with GFP at the carboxy terminus. The synthetic chimeric gene (native-phylloplanin-GFP) was placed between the modified Mirabilis mosaic virus full-length transcript promoter with duplicated enhancer domains and the terminator sequence from the rbcSE9 gene. The chimeric gene, expressed in transgenic tobacco, was stably inherited in successive plant generations as shown by molecular characterization, GFP quantification, and confocal fluorescent microscopy. Transgenic plants were morphologically similar to wild-type plants and showed no deleterious effects due to transgene expression. Blue mold-sensitivity assays of tobacco lines were performed by applying P. tabacina sporangia to the upper leaf surface. Transgenic lines expressing the fused synthetic native-phyllopanin-GFP gene in the leaf apoplast showed resistance to infection. Our results demonstrate that in vivo expression of a synthetic fused native-phylloplanin-GFP gene in plants can potentially achieve natural protection against microbial plant pathogens, including P. tabacina in tobacco. Dipak K. Sahoo, Sumita Raha, James T. Hall, and Indu B. Maiti Copyright © 2014 Dipak K. Sahoo et al. All rights reserved. Evaluation the Surface Antigen of the Salmonella typhimurium ATCC 14028 Ghosts Prepared by “SLRP” Wed, 19 Mar 2014 08:20:18 +0000 Recently, bacterial ghosts (BGs) were prepared using a protocol based on critical chemical concentrations. It has been given the name “sponge like” (SL) protocol and used in its reduced form “sponge like reduced protocol” (SLRP). While specific antibody for Salmonella is available on the market under the commercial names (of some kits) such as Febrile Antigen Kit (N.S. BIO-TEC), we used the described Kit to investigate the validity of the SLRP. In this study, using SLRP we succeeded to prepare STGs with correct surface antigens could interact with their specific antibodies. Additionally the study has included oral vaccination with STGs with challenge test. The rats serums have been evaluated against both of the O and H antigens. The antigen-antibody interaction (agglutination) results of both the SLRP and the animal experiments prove that we have correct STGs able to immunize the rats against viable Salmonella. STGs could be used as vaccine and as adjuvant and in the antibodies and in the diagnostic kits production. This study is an additional step for the establishment of correct BGs for immunological purposes. Amara A. Amro, Ahmed J. Neama, Ahmed Hussein, Emad A. Hashish, and Salah A. Sheweita Copyright © 2014 Amara A. Amro et al. All rights reserved. Gene Technology for Papaya Ringspot Virus Disease Management Mon, 17 Mar 2014 00:00:00 +0000 Papaya (Carica papaya) is severely damaged by the papaya ringspot virus (PRSV). This review focuses on the development of PRSV resistant transgenic papaya through gene technology. The genetic diversity of PRSV depends upon geographical distribution and the influence of PRSV disease management on a sequence of PRSV isolates. The concept of pathogen-derived resistance has been employed for the development of transgenic papaya, using a coat protein-mediated, RNA-silencing mechanism and replicase gene-mediated transformation for effective PRSV disease management. The development of PRSV-resistant papaya via post-transcriptional gene silencing is a promising technology for PRSV disease management. PRSV-resistant transgenic papaya is environmentally safe and has no harmful effects on human health. Recent studies have revealed that the success of adoption of transgenic papaya depends upon the application, it being a commercially viable product, bio-safety regulatory issues, trade regulations, and the wider social acceptance of the technology. This review discusses the genome and the genetic diversity of PRSV, host range determinants, molecular diagnosis, disease management strategies, the development of transgenic papaya, environmental issues, issues in the adoption of transgenic papaya, and future directions for research. Md. Abul Kalam Azad, Latifah Amin, and Nik Marzuki Sidik Copyright © 2014 Md. Abul Kalam Azad et al. All rights reserved. Improved Design of Anaerobic Digesters for Household Biogas Production in Indonesia: One Cow, One Digester, and One Hour of Cooking per Day Thu, 13 Mar 2014 14:13:04 +0000 A government-sponsored initiative in Indonesia to design and implement low-cost anaerobic digestion systems resulted in 21 full-scale systems with the aim to satisfy the cooking fuel demands of rural households owning at least one cow. The full-scale design consisted of a 0.3 m diameter PVC pipe, which was operated as a conventional plug-flow system. The system generated enough methane to power a cooking stove for ∼1 h. However, eventual clogging from solids accumulation inside the bioreactor proved to be a major drawback. Here, we improved the digester configuration to remedy clogging while maintaining system performance. Controlled experiments were performed using four 9-L laboratory-scale digesters operated at a temperature of °C, a volatile solids loading rate of 2.0 g VS·L−1·day−1, and a 21-day hydraulic retention time. Two of the digesters were replicates of the original design (control digesters), while the other two digesters included internal mixing or effluent recycle (experimental digesters). The performance of each digester was compared based on methane yields, VS removal efficiencies, and steady-state solids concentrations during an operating period of 311 days. Statistical analyses revealed that internal mixing and effluent recycling resulted in reduced solids accumulation compared to the controls without diminishing methane yields or solids removal efficiencies. Joseph G. Usack, Wiratni Wiratni, and Largus T. Angenent Copyright © 2014 Joseph G. Usack et al. All rights reserved. Determination of Suitable Microspore Stage and Callus Induction from Anthers of Kenaf (Hibiscus cannabinus L.) Thu, 13 Mar 2014 08:37:05 +0000 Kenaf (Hibiscus cannabinus L.) is one of the important species of Hibiscus cultivated for fiber. Availability of homozygous parent lines is prerequisite to the use of the heterosis effect reproducible in hybrid breeding. The production of haploid plants by anther culture followed by chromosome doubling can be achieved in short period compared with inbred lines by conventional method that requires self pollination of parent material. In this research, the effects of the microspore developmental stage, time of flower collection, various pretreatments, different combinations of hormones, and culture condition on anther culture of KB6 variety of Kenaf were studied. Young flower buds with immature anthers at the appropriate stage of microspore development were sterilized and the anthers were carefully dissected from the flower buds and subjected to various pretreatments and different combinations of hormones like NAA, 2,4-D, Kinetin, BAP, and TDZ to induce callus. The best microspore development stage of the flower buds was about 6–8 mm long collected 1-2 weeks after flower initiation. At that stage, the microspores were at the uninucleate stage which was suitable for culture. The best callus induction frequency was 90% in the optimized semisolid MS medium fortified with 3.0 mg/L BAP + 3.0 mg/L NAA. Ahmed Mahmood Ibrahim, Fatimah Binti Kayat, Zeti Ermiena Surya Mat Hussin, Dwi Susanto, and Mohammed Ariffulah Copyright © 2014 Ahmed Mahmood Ibrahim et al. All rights reserved. An Efficient In Vitro Plantlet Regeneration from Shoot Tip Cultures of Curculigo latifolia, a Medicinal Plant Thu, 27 Feb 2014 09:16:09 +0000 A procedure was developed for in vitro propagation of Curculigo latifolia through shoot tip culture. Direct regeneration and indirect scalp induction of Curculigo latifolia were obtained from shoot tip grown on MS medium supplemented with different concentrations and combinations of thidiazuron and indole-3-butyric acid. Maximum response for direct regeneration in terms of percentage of explants producing shoot, shoot number, and shoot length was obtained on MS medium supplemented with combination of thidiazuron (0.5 mg L−1) and indole-3-butyric acid (0.25 mg L−1) after both 10 and 14 weeks of cultures. Indole-3-butyric acid in combination with thidiazuron exhibited a synergistic effect on shoot regeneration. The shoot tips were able to induce maximum scalp from basal end of explants on the medium with 2 mg L−1 thidiazuron. Cultures showed that shoot number, shoot length, and scalp size increased significantly after 14 weeks of culture. Transferring of the shoots onto the MS medium devoid of growth regulators resulted in the highest percentage of root induction and longer roots, while medium supplemented with 0.25 mg L−1 IBA produced more numbers of roots. Nahid Babaei, Nur Ashikin Psyquay Abdullah, Ghizan Saleh, and Thohirah Lee Abdullah Copyright © 2014 Nahid Babaei et al. All rights reserved. Modeling the Differences in Biochemical Capabilities of Pseudomonas Species by Flux Balance Analysis: How Good Are Genome-Scale Metabolic Networks at Predicting the Differences? Mon, 24 Feb 2014 07:54:10 +0000 To date, several genome-scale metabolic networks have been reconstructed. These models cover a wide range of organisms, from bacteria to human. Such models have provided us with a framework for systematic analysis of metabolism. However, little effort has been put towards comparing biochemical capabilities of closely related species using their metabolic models. The accuracy of a model is highly dependent on the reconstruction process, as some errors may be included in the model during reconstruction. In this study, we investigated the ability of three Pseudomonas metabolic models to predict the biochemical differences, namely, iMO1086, iJP962, and iSB1139, which are related to P. aeruginosa PAO1, P. putida KT2440, and P. fluorescens SBW25, respectively. We did a comprehensive literature search for previous works containing biochemically distinguishable traits over these species. Amongst more than 1700 articles, we chose a subset of them which included experimental results suitable for in silico simulation. By simulating the conditions provided in the actual biological experiment, we performed case-dependent tests to compare the in silico results to the biological ones. We found out that iMO1086 and iJP962 were able to predict the experimental data and were much more accurate than iSB1139. Parizad Babaei, Tahereh Ghasemi-Kahrizsangi, and Sayed-Amir Marashi Copyright © 2014 Parizad Babaei et al. All rights reserved. Catalyzed Ester Synthesis Using Candida rugosa Lipase Entrapped by Poly(N-isopropylacrylamide-co-itaconic Acid) Hydrogel Thu, 20 Feb 2014 15:50:37 +0000 This study reports the synthesis of polymeric matrices based on N-isopropylacrylamide and itaconic acid and its application for immobilization of lipase from Candida rugosa. The lipase was immobilized by entrapment method. Free and immobilized lipase activities, pH and temperature optima, and storage stability were investigated. The optimum temperature for free and entrapped lipase was found to be 40 and 45°C, while the optimum pH was observed at pH 7 and 8, respectively. Both hydrolytic activity in an aqueous medium and esterolytic activity in an organic medium have been evaluated. Maximum reaction rate () and Michaelis-Menten constants () were also determined for immobilized lipase. Storage stability of lipase was increased as a result of immobilization process. Furthermore, the operational stability and reusability of the immobilized lipase in esterification reaction have been studied, and it was observed that after 10 cycles, the residual activity for entrapped lipase was as high as 50%, implying that the developed hydrogel and immobilized system could provide a promising solution for the flavor ester synthesis at the industrial scale. Nikola Milašinović, Sonja Jakovetić, Zorica Knežević-Jugović, Nedeljko Milosavljević, Marija Lučić, Jovanka Filipović, and Melina Kalagasidis Krušić Copyright © 2014 Nikola Milašinović et al. All rights reserved. Hypocholesterolemic Activity of Monascus Fermented Product in the Absence of Monacolins with Partial Purification for Functional Food Applications Thu, 20 Feb 2014 09:24:51 +0000 Hypercholesterolemia is one of the most common chronic diseases in human. Along with chemical therapy traditional medication is used as hypocholesterolemic remedy, however, with unfavorable side effects. Recently, Monascus fermented product (MFP) has become a popular hypocholesterolemic natural supplement. In the present study, the hypocholesterolemic activity of Monascus purpureus FTC5391 fermented product ethanolic extract (MFPe) was investigated in hypercholesterolemic rats. Results showed that MFPe not only reduced the serum total cholesterol (TC), LDL-C, TG concentration, and TC/HDL-C ratio but also increased the HDL-C. Further, solid phase extraction (SPE) was carried out to obtain the hypocholesterolemic bioactive fraction. The high polar fraction of SPE increased the HDL-C (42%) and decreased the TC (53.3%), LDL-C (47%), and TG (50.7%) levels as well as TC/HDL-C ratio (69.1%) in serum. The GC-MS results of the active fraction revealed two main compounds, isosorbide and erythritol, which act as coronary vasodilator compounds. Zahra Ajdari, Maaruf Abd Ghani, Mohd Khan Ayob, Saadi Bayat, Mazlin Mokhtar, Sahar Abbasiliasi, Anahita Khoramnia, Heshu Sulaiman Rahman, Parvaneh Mehrbod, Daniel Ajdari, and Arbakariya B. Ariff Copyright © 2014 Zahra Ajdari et al. All rights reserved. Activity of a Carboxyl-Terminal Truncated Form of Catechol 2,3-Dioxygenase from Planococcus sp. S5 Thu, 13 Feb 2014 00:00:00 +0000 Catechol 2,3-dioxygenases (C23Os, E.C. are two domain enzymes that catalyze degradation of monoaromatic hydrocarbons. The catalytically active C-domain of all known C23Os comprises ferrous ion ligands as well as residues forming active site pocket. The aim of this work was to examine and discuss the effect of nonsense mutation at position 289 on the activity of catechol 2,3-dioxygenase from Planococcus strain. Although the mutant C23O showed the same optimal temperature for activity as the wild-type protein (35°C), it exhibited activity slightly more tolerant to alkaline pH. Mutant enzyme exhibited also higher affinity to catechol as a substrate. Its Km (66.17 µM) was approximately 30% lower than that of wild-type enzyme. Interestingly, removal of the C-terminal residues resulted in 1.5- to 1.8-fold increase in the activity of C23OB61 against 4-methylcatechol and 4-chlorocatechol, respectively, while towards catechol the activity of the protein dropped to about 80% of that of the wild-type enzyme. The results obtained may facilitate the engineering of the C23O for application in the bioremediation of polluted areas. Katarzyna Hupert-Kocurek, Danuta Wojcieszyńska, and Urszula Guzik Copyright © 2014 Katarzyna Hupert-Kocurek et al. All rights reserved. Antioxidant Capacity and Total Phenolic Content in Fruit Tissues from Accessions of Capsicum chinense Jacq. (Habanero Pepper) at Different Stages of Ripening Tue, 11 Feb 2014 00:00:00 +0000 In the past few years, there has been a renewed interest in studying a wide variety of food products that show beneficial effects on human health. Capsicum is an important agricultural crop, not only because its economic importance, but also for the nutritional values of its pods, mainly due to the fact that they are an excellent source of antioxidant compounds, and also of specific constituents such as the pungent capsaicinoids localized in the placental tissue. This current study was designed to evaluate the antioxidant capacity and total phenolic contents from fruits tissues of two Capsicum chinense accessions, namely, Chak k’an-iik (orange) and MR8H (red), at contrasting maturation stages. Results showed that red immature placental tissue, with a Trolox equivalent antioxidant capacity (TEAC) value of 55.59 μmols TE g−1 FW, exhibited the strongest total antioxidant capacity using both the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and the CUPRAC methods. Placental tissue also had the highest total phenolic content (27 g GAE 100 g−1 FW). The antioxidant capacity of Capsicum was directly related to the total amount of phenolic compounds detected. In particular, placentas had high levels of capsaicinoids, which might be the principal responsible for their strong antioxidant activities. Lizbeth A. Castro-Concha, Jemina Tuyub-Che, Angel Moo-Mukul, Felipe A. Vazquez-Flota, and Maria L. Miranda-Ham Copyright © 2014 Lizbeth A. Castro-Concha et al. All rights reserved. Next Generation Sequencing: Potential and Application in Drug Discovery Wed, 05 Feb 2014 07:46:21 +0000 The world has now entered into a new era of genomics because of the continued advancements in the next generation high throughput sequencing technologies, which includes sequencing by synthesis-fluorescent in situ sequencing (FISSEQ), pyrosequencing, sequencing by ligation using polony amplification, supported oligonucleotide detection (SOLiD), sequencing by hybridization along with sequencing by ligation, and nanopore technology. Great impacts of these methods can be seen for solving the genome related problems of plant and animal kingdom that will open the door of a new era of genomics. This may ultimately overcome the Sanger sequencing that ruled for 30 years. NGS is expected to advance and make the drug discovery process more rapid. Navneet Kumar Yadav, Pooja Shukla, Ankur Omer, Shruti Pareek, and R. K. Singh Copyright © 2014 Navneet Kumar Yadav et al. All rights reserved. High Efficiency Direct Shoot Organogenesis from Leaf Segments of Aerva lanata (L.) Juss. Ex Schult by Using Thidiazuron Tue, 04 Feb 2014 12:12:32 +0000 An efficient protocol for direct shoot organogenesis has been developed for the medicinal plant Aerva lanata (L.) Juss. ex Schult. Regeneration was achieved from leaf segments of 20 days old in vitro plantlets raised on Murashige and Skoog (MS) medium containing 0.25–2.0 mg L−1 thiadiazuron (TDZ), 3% sucrose, and 0.8% agar. After 21 days of culture incubation, maximum number of shoot organogenesis (23.6 ± 0.16) was obtained on medium containing 1.0 mg L−1 TDZ. The shoots were able to produce in vitro flowers on medium containing 1.0 mg L−1 TDZ in combination with 0.25–0.5 mg L−1  α-naphthaleneacetic acid (NAA). Histological observation showed that the epidermal cells of the leaf explants exhibited continuous cell division led to formation of numerous dome shaped meristematic protrusions and subsequently developed into adventitious shoots. Upon transfer of shootlets to half strength MS medium containing 1.0 mg L−1 indole-3-butyric acid (IBA), around 86% of the regenerated shoots formed roots and plantlets. Rooted plants were hardened and successfully established in the soil at the survival rate of 92%. The regeneration protocol developed in this study provides an important method of micropropagation of this plant. Furthermore, this protocol may be used for a large scale production of its medicinally active compounds and genetic transformations for further improvement. K. Varutharaju, C. Soundar Raju, C. Thilip, A. Aslam, and A. Shajahan Copyright © 2014 K. Varutharaju et al. All rights reserved. Comparative Study of Antioxidant Status in Androgenic Embryos of Aesculus hippocastanum and Aesculus flava Mon, 03 Feb 2014 09:20:11 +0000 In vivo (leaves and seed embryos) and in vitro (androgenic embryos) antioxidant scavenging activity of Aesculus hippocastanum and Aesculus flava medical plants was examined. Here we report antioxidant enzyme activities of superoxide dismutase, catalase, guaiacol peroxidase and glutathione peroxidase, reduced glutathione quantity, flavonoids, soluble protein contents, quantities of malondialdehyde, and •OH radical presence in the investigated plant samples. Total antioxidant capacity of all the samples of A. hippocastanum and A. flava was determined using FRAP, DPPH, and NO• radical scavenger capacity. The leaves of A. flava collected from the botanical garden exhibited stronger antioxidant activity (higher activities of SOD, and higher quantities of GSH, TSH, TPC, and scavenging abilities of DPPH and NO•, and higher FRAP values and lowest quantities of •OH and MDA) than in vitro obtained cultures. However, the leaves of A. flava showed higher antioxidant activity than the leaves of A. hippocastanum, and therefore they have a stronger tolerance of oxidative stress. Androgenic embryos of both species had low amount of antioxidants due to controlled in vitro environmental conditions (T, photoperiod, humidity, nutritive factors, and pathogen-free). Our results confirmed that we found optimal in vitro conditions for producing androgenic embryos of both Aesculus species. Also, we assume that horse chestnut androgenic embryos can be used as an alternative source for large-scale aescin production. Dubravka Štajner, Boris M. Popović, Dušica Ćalić, and Marijana Štajner Copyright © 2014 Dubravka Štajner et al. All rights reserved. Optimal Immobilization of β-Galactosidase onto κ-Carrageenan Gel Beads Using Response Surface Methodology and Its Applications Sun, 02 Feb 2014 11:36:09 +0000 β-Galactosidase (β-gal) was immobilized by covalent binding on novel κ-carrageenan gel beads activated by two-step method; the gel beads were soaked in polyethyleneimine followed by glutaraldehyde. 22 full-factorial central composite experiment designs were employed to optimize the conditions for the maximum enzyme loading efficiency. 11.443 U of enzyme/g gel beads was achieved by soaking 40 units of enzyme with the gel beads for eight hours. Immobilization process increased the pH from 4.5 to 5.5 and operational temperature from 50 to 55°C compared to the free enzyme. The apparent after immobilization was 61.6 mM compared to 22.9 mM for free enzyme. Maximum velocity was 131.2 μmol·min−1 while it was 177.1 μmol·min−1 for free enzyme. The full conversion experiment showed that the immobilized enzyme form is active as that of the free enzyme as both of them reached their maximum 100% relative hydrolysis at 4 h. The reusability test proved the durability of the κ-carrageenan beads loaded with β-galactosidase for 20 cycles with retention of 60% of the immobilized enzyme activity to be more convenient for industrial uses. Magdy M. Elnashar, Ghada E. Awad, Mohamed E. Hassan, Mohamed S. Mohy Eldin, Bakry M. Haroun, and Ahmed I. El-Diwany Copyright © 2014 Magdy M. Elnashar et al. All rights reserved. Organic Solvent Tolerant Lipases and Applications Sun, 02 Feb 2014 07:31:46 +0000 Lipases are a group of enzymes naturally endowed with the property of performing reactions in aqueous as well as organic solvents. The esterification reactions using lipase(s) could be performed in water-restricted organic media as organic solvent(s) not only improve(s) the solubility of substrate and reactant in reaction mixture but also permit(s) the reaction in the reverse direction, and often it is easy to recover the product in organic phase in two-phase equilibrium systems. The use of organic solvent tolerant lipase in organic media has exhibited many advantages: increased activity and stability, regiospecificity and stereoselectivity, higher solubility of substrate, ease of products recovery, and ability to shift the reaction equilibrium toward synthetic direction. Therefore the search for organic solvent tolerant enzymes has been an extensive area of research. A variety of fatty acid esters are now being produced commercially using immobilized lipase in nonaqueous solvents. This review describes the organic tolerance and industrial application of lipases. The main emphasis is to study the nature of organic solvent tolerant lipases. Also, the potential industrial applications that make lipases the biocatalysts of choice for the present and future have been presented. Shivika Sharma and Shamsher S. Kanwar Copyright © 2014 Shivika Sharma and Shamsher S. Kanwar. All rights reserved. Kinetic Studies on Fermentative Production of Biofuel from Synthesis Gas Using Clostridium ljungdahlii Thu, 30 Jan 2014 08:31:52 +0000 The intrinsic growth, substrate uptake, and product formation biokinetic parameters were obtained for the anaerobic bacterium, Clostridium ljungdahlii, grown on synthesis gas in various pressurized batch bioreactors. A dual-substrate growth kinetic model using Luong for CO and Monod for H2 was used to describe the growth kinetics of the bacterium on these substrates. The maximum specific growth rate ( = 0.195 h−1) and Monod constants for CO ( = 0.855 atm) and H2 ( = 0.412 atm) were obtained. This model also accommodated the CO inhibitory effects on cell growth at high CO partial pressures, where no growth was apparent at high dissolved CO tensions ( atm). The Volterra model, Andrews, and modified Gompertz were, respectively, adopted to describe the cell growth, substrate uptake rate, and product formation. The maximum specific CO uptake rate ( = 34.364 mmol//h), CO inhibition constant ( = 0.601 atm), and maximum rate of ethanol ( = 0.172 mmol/L/h at = 0.598 atm) and acetate ( = 0.096 mmol/L/h at = 0.539 atm) production were determined from the applied models. Maedeh Mohammadi, Abdul Rahman Mohamed, Ghasem D. Najafpour, Habibollah Younesi, and Mohamad Hekarl Uzir Copyright © 2014 Maedeh Mohammadi et al. All rights reserved. Influence of Cytokinins in Combination with GA3 on Shoot Multiplication and Elongation of Tea Clone Iran 100 (Camellia sinensis (L.) O. Kuntze) Tue, 28 Jan 2014 11:14:34 +0000 The use of in vitro culture has been accepted as an efficient technique for clonal propagation of many woody plants. In the present research, we report the results of a number of experiments aimed at optimizing micropropagation protocol for tea (Camellia sinensis (L.) O. Kuntze) (clone Iran 100) using nodal segments as the explant. The effect of different combinations and concentrations of plant growth regulators (PGR) (BAP, TDZ, GA3) on shoot multiplication and elongation was assessed. The influence of exposure to IBA in liquid form prior to transfer to solid media on rooting of tea microshoots was investigated. The results of this study showed that the best treatment for nodal segment multiplication in terms of the number of shoot per explant and shoot elongation was obtained using 3 mg/L BAP in combination with 0.5 mg/L GA3. TDZ was found to be inappropriate for multiplication of tea clone Iran 100 as it resulted in hyperhydricity especially at concentrations higher than 0.05 mg/L. Healthy shoots treated with 300 mg/L IBA for 30 min followed by transfer to 1/2 strength MS medium devoid of PGR resulted in 72.3% of shoots producing roots and upon transferring them to acclimatization chamber 65% survival was obtained prior to field transfer. Reza Azadi Gonbad, Uma Rani Sinniah, Maheran Abdul Aziz, and Rosfarizan Mohamad Copyright © 2014 Reza Azadi Gonbad et al. All rights reserved. Chemoenzymatic Epoxidation of Alkenes and Reusability Study of the Phenylacetic Acid Mon, 27 Jan 2014 12:34:59 +0000 Here, we focused on a simple enzymatic epoxidation of alkenes using lipase and phenylacetic acid. The immobilised Candida antarctica lipase B, Novozym 435 was used to catalyse the formation of peroxy acid instantly from hydrogen peroxide (H2O2) and phenylacetic acid. The peroxy phenylacetic acid generated was then utilised directly for in situ oxidation of alkenes. A variety of alkenes were oxidised with this system, resulting in 75–99% yield of the respective epoxides. On the other hand, the phenylacetic acid was recovered from the reaction media and reused for more epoxidation. Interestingly, the waste phenylacetic acid had the ability to be reused for epoxidation of the 1-nonene to 1-nonene oxide, giving an excellent yield of 90%. Emilia Abdulmalek, Mahashanon Arumugam, Hanis Nabillah Mizan, Mohd. Basyaruddin Abdul Rahman, Mahiran Basri, and Abu Bakar Salleh Copyright © 2014 Emilia Abdulmalek et al. All rights reserved. Assessment of Total Phenolic, Antioxidant, and Antibacterial Activities of Passiflora Species Tue, 21 Jan 2014 12:51:29 +0000 This study focused on total phenolic content (TPC) and antioxidant and antibacterial activities of the leaves and stems of Passiflora quadrangularis, P. maliformis, and P. edulis extracted using three solvents: petroleum ether, acetone, and methanol. The maximum extraction yields of antioxidant components from the leaves and stems were isolated using methanol extracts of P. edulis (24.28%) and P. quadrangularis (9.76%), respectively. Among the leaf extracts, the methanol extract of P. maliformis had the significantly highest TPC and the strongest antioxidant activity, whereas among the stem extracts, the methanol extract of P. quadrangularis showed the highest phenolic amount and possessed the strongest antioxidant activity. The antibacterial properties of the Passiflora species were tested using the disc diffusion method against 10 human pathogenic bacteria. The largest inhibition zone was observed for the methanol extract of P. maliformis against B. subtilis. Generally, extracts from the Passiflora species exhibit distinct inhibition against Gram-positive but not Gram-negative bacteria. Based on the generated biplot, three clusters of bacteria were designated according to their performance towards the tested extracts. The present study revealed that methanol extracts of the Passiflora contain constituents with significant phenolic, antioxidant, and antibacterial properties for pharmaceutical and nutraceutical uses. Shiamala Devi Ramaiya, Japar Sidik Bujang, and Muta Harah Zakaria Copyright © 2014 Shiamala Devi Ramaiya et al. All rights reserved. The Enzymatic Decolorization of Textile Dyes by the Immobilized Polyphenol Oxidase from Quince Leaves Tue, 21 Jan 2014 12:26:28 +0000 Water pollution due to release of industrial wastewater has already become a serious problem in almost every industry using dyes to color its products. In this work, polyphenol oxidase enzyme from quince (Cydonia Oblonga) leaves immobilized on calcium alginate beads was used for the successful and effective decolorization of textile industrial effluent. Polyphenol oxidase (PPO) enzyme was extracted from quince (Cydonia Oblonga) leaves and immobilized on calcium alginate beads. The kinetic properties of free and immobilized PPO were determined. Quince leaf PPO enzyme stability was increased after immobilization. The immobilized and free enzymes were employed for the decolorization of textile dyes. The dye solutions were prepared in the concentration of 100 mg/L in distilled water and incubated with free and immobilized quince (Cydonia Oblonga) leaf PPO for one hour. The percent decolorization was calculated by taking untreated dye solution. Immobilized PPO was significantly more effective in decolorizing the dyes as compared to free enzyme. Our results showed that the immobilized quince leaf PPO enzyme could be efficiently used for the removal of synthetic dyes from industrial effluents. Gulnur Arabaci and Ayse Usluoglu Copyright © 2014 Gulnur Arabaci and Ayse Usluoglu. All rights reserved. Molecular Divergence and Species Delimitation of the Cultivated Oyster Mushrooms: Integration of IGS1 and ITS Tue, 21 Jan 2014 08:51:13 +0000 Identification of edible mushrooms particularly Pleurotus genus has been restricted due to various obstacles. The present study attempted to use the combination of two variable regions of IGS1 and ITS for classifying the economically cultivated Pleurotus species. Integration of the two regions proved a high ability that not only could clearly distinguish the species but also served sufficient intraspecies variation. Phylogenetic tree (IGS1 + ITS) showed seven distinct clades, each clade belonging to a separate species group. Moreover, the species differentiation was tested by AMOVA and the results were reconfirmed by presenting appropriate amounts of divergence (91.82% among and 8.18% within the species). In spite of achieving a proper classification of species by combination of IGS1 and ITS sequences, the phylogenetic tree showed the misclassification of the species of P. nebrodensis and P. eryngii var. ferulae with other strains of P. eryngii. However, the constructed median joining (MJ) network could not only differentiate between these species but also offer a profound perception of the species’ evolutionary process. Eventually, due to the sufficient variation among and within species, distinct sequences, simple amplification, and location between ideal conserved ribosomal genes, the integration of IGS1 and ITS sequences is recommended as a desirable DNA barcode. Farhat Ahmadi Avin, Subha Bhassu, Yee Shin Tan, Pedram Shahbazi, and Sabaratnam Vikineswary Copyright © 2014 Farhat Ahmadi Avin et al. All rights reserved. Community Structure and Succession Regulation of Fungal Consortia in the Lignocellulose-Degrading Process on Natural Biomass Sun, 19 Jan 2014 00:00:00 +0000 The study aims to investigate fungal community structures and dynamic changes in forest soil lignocellulose-degrading process. rRNA gene clone libraries for the samples collected in different stages of lignocellulose degradation process were constructed and analyzed. A total of 26 representative RFLP types were obtained from original soil clone library, including Mucoromycotina (29.5%), unclassified Zygomycetes (33.5%), Ascomycota (32.4%), and Basidiomycota (4.6%). When soil accumulated with natural lignocellulose, 16 RFLP types were identified from 8-day clone library, including Basidiomycota (62.5%), Ascomycota (36.1%), and Fungi incertae sedis (1.4%). After enrichment for 15 days, identified 11 RFLP types were placed in 3 fungal groups: Basidiomycota (86.9%), Ascomycota (11.5%), and Fungi incertae sedis (1.6%). The results showed richer, more diversity and abundance fungal groups in original forest soil. With the degradation of lignocellulose, fungal groups Mucoromycotina and Ascomycota decreased gradually, and wood-rotting fungi Basidiomycota increased and replaced the opportunist fungi to become predominant group. Most of the fungal clones identified in sample were related to the reported lignocellulose-decomposing strains. Understanding of the microbial community structure and dynamic change during natural lignocellulose-degrading process will provide us with an idea and a basis to construct available commercial lignocellulosic enzymes or microbial complex. Baoyu Tian, Chunxiang Wang, Ruirui Lv, Junxiong Zhou, Xin Li, Yi Zheng, Xiangyu Jin, Mengli Wang, Yongxia Ye, Xinyi Huang, and Ping Liu Copyright © 2014 Baoyu Tian et al. All rights reserved. Novel Glycolipids Synthesized Using Plant Essential Oils and Their Application in Quorum Sensing Inhibition and as Antibiofilm Agents Tue, 14 Jan 2014 11:40:09 +0000 Essential oils (EOs) form an important part of traditional medicine so their anti-microbial and, in the recent past, antiquorum sensing activity has been well studied. However it is likely that due to their hydrophobic nature and reduced solubility in aqueous environments full potential of their activity cannot be realized. hence it is only rational to formulate a process to make these molecules more polar in nature. The present paper reports synthesis of sophorolipids using 12 different essential oils as substrates, thus providing surfactant-like properties to these EOs. The synthesis protocol makes the use of Candida bombicola ATCC 22214 as producer organism. The production process required 7 days of incubation at 28°C and 180 rpm. Preliminary characterization of the synthesized essential oil sophorolipids (EOSLs) was performed using thin layer chromatography (TLC) and Fourier transform infrared spectroscopy (FTIR). Additionally, essential oils that were incapable of mediating quorum sensing inhibition (QSI) on their own became potent quorum sensing inhibitors upon conversion into their corresponding EOSLs. Antibiofilm potential of these EOSLs was also demonstrated using V. cholerae as test organism. Use of essential oils as substrates for glycolipid synthesis has not been attempted previously, and hence this is the first report. Ruchira Mukherji and Asmita Prabhune Copyright © 2014 Ruchira Mukherji and Asmita Prabhune. All rights reserved. Silicon Promotes Adventitious Shoot Regeneration and Enhances Salinity Tolerance of Ajuga multiflora Bunge by Altering Activity of Antioxidant Enzyme Thu, 09 Jan 2014 13:19:47 +0000 We investigated the effect of Si concentration on shoot regeneration and salinity tolerance of Ajuga multiflora. Addition of Si to the shoot induction medium significantly increased the frequency of shoot induction. The average number of shoots regenerated per explant decreased on the medium containing NaCl alone, while there was less decrease when the shoot induction medium was supplemented with both NaCl and Si. The shoot induction percentage increased linearly with increasing concentration of Si in the NaCl containing medium. Addition of Si to the shoot induction medium significantly increased SOD, POD, APX, and CAT activity in regenerated shoot buds as compared with the control. The inclusion of Si to the NaCl containing medium significantly increased the SOD activity in leaves and roots, while it decreased POD, APX, and CAT activity in both organs. Scanning electron microscopic analysis showed that there are no distinct differences in the structure of stomata between the control and Si-treated plants. However, NaCl treatment significantly affected the structure and number of stomata as compared to the control. Wavelength dispersive X-ray analysis confirmed the high Si deposition in trichomes of plants grown in the Si containing medium but not in plants grown in the medium without Si. Iyyakkannu Sivanesan and Byoung Ryong Jeong Copyright © 2014 Iyyakkannu Sivanesan and Byoung Ryong Jeong. All rights reserved. Neural and Hybrid Modeling: An Alternative Route to Efficiently Predict the Behavior of Biotechnological Processes Aimed at Biofuels Obtainment Sun, 05 Jan 2014 12:33:54 +0000 The present paper was aimed at showing that advanced modeling techniques, based either on artificial neural networks or on hybrid systems, might efficiently predict the behavior of two biotechnological processes designed for the obtainment of second-generation biofuels from waste biomasses. In particular, the enzymatic transesterification of waste-oil glycerides, the key step for the obtainment of biodiesel, and the anaerobic digestion of agroindustry wastes to produce biogas were modeled. It was proved that the proposed modeling approaches provided very accurate predictions of systems behavior. Both neural network and hybrid modeling definitely represented a valid alternative to traditional theoretical models, especially when comprehensive knowledge of the metabolic pathways, of the true kinetic mechanisms, and of the transport phenomena involved in biotechnological processes was difficult to be achieved. Stefano Curcio, Alessandra Saraceno, Vincenza Calabrò, and Gabriele Iorio Copyright © 2014 Stefano Curcio et al. All rights reserved. Association Analysis of SSR Markers with Phenology, Grain, and Stover-Yield Related Traits in Pearl Millet (Pennisetum glaucum (L.) R. Br.) Thu, 02 Jan 2014 08:50:50 +0000 Pearl millet is a staple food crop for millions of people living in the arid and semi-arid tropics. Molecular markers have been used to identify genomic regions linked to traits of interest by conventional QTL mapping and association analysis. Phenotypic recurrent selection is known to increase frequencies of favorable alleles and decrease those unfavorable for the traits under selection. This study was undertaken (i) to quantify the response to recurrent selection for phenotypic traits during breeding of the pearl millet open-pollinated cultivar “CO (Cu) 9” and its four immediate progenitor populations and (ii) to assess the ability of simple sequence repeat (SSR) marker alleles to identify genomic regions linked to grain and stover yield-related traits in these populations by association analysis. A total of 159 SSR alleles were detected across 34 selected single-copy SSR loci. SSR marker data revealed presence of subpopulations. Association analysis identified genomic regions associated with flowering time located on linkage group (LG) 6 and plant height on LG4, LG6, and LG7. Marker alleles on LG6 were associated with stover yield, and those on LG7 were associated with grain yield. Findings of this study would give an opportunity to develop marker-assisted recurrent selection (MARS) or marker-assisted population improvement (MAPI) strategies to increase the rate of gain for pearl millet populations undergoing recurrent selection. Baskaran Kannan, Senthilvel Senapathy, Arcot Gajaraj Bhasker Raj, Subhash Chandra, Arunachalam Muthiah, Arun Prabhu Dhanapal, and Charles Thomas Hash Copyright © 2014 Baskaran Kannan et al. All rights reserved. Expression and Promoter Analysis of Six Heat Stress-Inducible Genes in Rice Thu, 26 Dec 2013 08:47:52 +0000 During the long evolutionary process, plant gradually formed a series of strategies and mechanisms to cope with stress environment such as drought, heat, cold, and high salinity. Six highly heat responsive genes were identified in rice by microarray data analysis. The qRT-PCR analysis confirmed that the expression of these six genes were highly heat inducible and moderately responded to salt stress, polyethylene glycol, and abscisic acid treatment, but little affected by cold treatment. Promoters of the three highly heat-inducible genes (OsHsfB2cp, PM19p, and Hsp90p) were used to drive GUS gene expression in rice. The results of the GUS gene expression, histochemical staining, and GUS activities in panicles and flag leaves of the transgenic rice plants confirmed high heat-induced GUS activities and moderate drought-induced activities. The three promoters exhibited similar high activity lever in rice leaf under heat, but OsHsfB2cp and PM19p showed much higher activities in panicles under heat stress. Our work confirmed that the OsHsfB2c and PM19 promoters were highly heat inducible and further characterization and reconstruction of cis-elements in their promoters could lead to the development of highly effective heat-inducible promoters for plant genetic engineering. Wirat Rerksiri, Xianwen Zhang, Hairong Xiong, and Xinbo Chen Copyright © 2013 Wirat Rerksiri et al. All rights reserved. Sustainable Power Generation in Continuous Flow Microbial Fuel Cell Treating Actual Wastewater: Influence of Biocatalyst Type on Electricity Production Wed, 25 Dec 2013 15:45:33 +0000 Microbial fuel cells (MFCs) have the potential to simultaneously treat wastewater for reuse and to generate electricity. This study mainly considers the performance of an upflow dual-chambered MFC continuously fueled with actual domestic wastewater and alternatively biocatalyzed with aerobic activated sludge and strain of Bacillus Subtilis. The behavior of MFCs during initial biofilm growth and characterization of anodic biofilm were studied. After 45 days of continuous operation, the biofilms on the anodic electrode were well developed. The performance of MFCs was mainly evaluated in terms of COD reductions and electrical power output. Results revealed that the COD removal efficiency was 84% and 90% and the stabilized power outputs were clearly observed achieving a maximum value of 120 and 270 mW/m2 obtained for MFCs inoculated with mixed cultures and Bacillus Subtilis strain, respectively. Zainab Z. Ismail and Ali Jwied Jaeel Copyright © 2013 Zainab Z. Ismail and Ali Jwied Jaeel. All rights reserved. Peroxidase(s) in Environment Protection Tue, 24 Dec 2013 09:51:29 +0000 Industrial discharges of untreated effluents into water bodies and emissions into air have deteriorated the quality of water and air, respectively. The huge amount of pollutants derived from industrial activities represents a threat for the environment and ecologic equilibrium. Phenols and halogenated phenols, polycyclic aromatic hydrocarbons (PAH), endocrine disruptive chemicals (EDC), pesticides, dioxins, polychlorinated biphenyls (PCB), industrial dyes, and other xenobiotics are among the most important pollutants. Peroxidases are enzymes that are able to transform a variety of compounds following a free radical mechanism, thereby yielding oxidized or polymerized products. The peroxidase transformation of these pollutants is accompanied by a reduction in their toxicity, due to loss of biological activity, reduction in the bioavailability, or the removal from aqueous phase, especially when the pollutant is found in water. The review describes the sources of peroxidases, the reactions catalyzed by them, and their applications in the management of pollutants in the environment. Neelam Bansal and Shamsher S. Kanwar Copyright © 2013 Neelam Bansal and Shamsher S. Kanwar. All rights reserved. Application of Response Surface Methodology for Optimizing Arginine Deiminase Production Medium for Enterococcus faecium sp. GR7 Tue, 17 Dec 2013 11:26:05 +0000 Arginine metabolism in Enterococcus faecium sp. GR7 was enhanced via arginine deiminase pathway. Process parameters including fermentation media and environmental conditions were optimized using independent experiments and response surface methodology (central composite design). Fermentation media (EAPM) were optimized using independent experiments which resulted in 4-fold increase in arginine deiminase specific activity as compared to basal medium. To further enhance arginine deiminase activity in E. faecium sp. GR7 and biomass production including a five-level central composite design (CCD) was employed to study the interactive effect of three-process variables. Response surface methodology suggested a quadratic model which was further validated experimentally where it showed approximately 15-fold increase in arginine metabolism (in terms of arginine deiminase specific activity) over basal medium. By solving the regression equation and analyzing the response surface cartons, optimal concentrations of the media components (g/L) were determined as arginine 20.0; tryptone 15.0; lactose 10.0; K2HPO4 3.0; NaCl 1.0, MnSO4 0.6 mM; Tween 80 1%; pH 6.0 for achieving specific arginine deiminase activity of 4.6 IU/mG with concomitant biomass production of 12.1 mg/L. The model is significant as the coefficient of determination () was 0.87 to 0.90 for all responses. Enhanced arginine deiminase yield from E. faecium, a GRAS lactic acid bacterial strain, is desirable to explore in vitro therapeutic potential of the arginine metabolizing E. faecium sp. GR7. Baljinder Kaur and Rajinder Kaur Copyright © 2013 Baljinder Kaur and Rajinder Kaur. All rights reserved. Chitinase Genes LbCHI31 and LbCHI32 from Limonium bicolor Were Successfully Expressed in Escherichia coli and Exhibit Recombinant Chitinase Activities Sat, 07 Dec 2013 13:49:42 +0000 The two chitinase genes, LbCHI31 and LbCHI32 from Limonium bicolor, were, respectively, expressed in Escherichia coli BL21 strain. The intracellular recombinant chitinases, inrCHI31 and inrCHI32, and the extracellular exrCHI31 and exrCHI32 could be produced into E. coli. The exrCHI31 and exrCHI32 can be secreted into extracellular medium. The optimal reaction condition for inrCHI31 was 5 mmol/L of Mn2+ at 40°C and pH 5.0 with an activity of 0.772 U using Alternaria alternata cell wall as substrate. The optimal condition of inrCHI32 was 5 mmol/L of Ba2+ at 45°C and pH 5.0 with an activity of 0.792 U using Valsa sordida cell wall as substrate. The optimal reaction condition of exrCHI31 was 5 mmol/L of Zn2+ at 40°C and pH 5.0, and the activity was 0.921 U using the A. alternata cell wall as substrate. Simultaneously, the optimal condition of exrCHI32 was 5 mmol/L of K+ at 45°C and pH 5.0, with V. sordida cell wall as the substrate, and the activity was 0.897 U. Furthermore, the activities of extracellular recombinant enzymes on fungal cell walls and compounds were generally higher than those of the intracellular recombinant enzymes. Recombinant exrCHI31 and exrCHI32 have better hydrolytic ability on cell walls of different fungi than synthetic chitins and obviously showed activity against A. alternata. Zhihua Liu, Ying Huang, Rongshu Zhang, Guiping Diao, Haijuan Fan, and Zhiying Wang Copyright © 2013 Zhihua Liu et al. All rights reserved. Early Pregnancy Diagnosis in Bovines: Current Status and Future Directions Thu, 05 Dec 2013 19:24:27 +0000 An early and accurate diagnosis of reproductive dysfunctions or aberrations is crucial to better reproductive management in livestock. High reproductive efficiency is a prerequisite for high life-time production in dairy animals. Early pregnancy diagnosis is key to shorten the calving interval through early identification of open animals and their timely treatment and rebreeding so as to maintain a postpartum barren interval close to 60 days. A buffalo, the most important dairy animal in the Indian subcontinent, is known for problems related to high calving interval, late puberty, and high incidence of anestrus. Lack of reliable cow-side early pregnancy diagnosis methods further aggravates the situation. Several methods of pregnancy diagnosis are being practiced in bovine species, yet none qualifies as the ideal pregnancy diagnosis method due to the inherent limitations of sensitivity, accuracy, specificity, speed, and ease of performing the test. The advancement of molecular techniques like proteomics and their applications in animal research has given a new hope to look for pregnancy biomarker molecules in these animals. This review attempts to examine common pregnancy diagnosis methods available for dairy animals, while assessing the usefulness of the modern technologies in detecting novel pregnancy markers and designing future strategies for research in this area. Ashok K. Balhara, Meenakshi Gupta, Surender Singh, Ashok K. Mohanty, and Inderjeet Singh Copyright © 2013 Ashok K. Balhara et al. All rights reserved. The Elicitation Effect of Pathogenic Fungi on Trichodermin Production by Trichoderma brevicompactum Thu, 05 Dec 2013 14:52:44 +0000 The effects of six species of phytopathogenic fungi mycelia as elicitors on trichodermin yield by Trichoderma brevicompactum were investigated. Neither nonviable nor viable mycelia of Botrytis cinerea, Alternaria solani, Colletotrichum lindemuthianum, and Thanatephorus cucumeris demonstrated any elicitation on the accumulation of trichodermin. However, the production of trichodermin was increased by the presence of viable/nonviable Rhizoctonia solani and Fusarium oxysporum mycelia. The strongest elicitation effect was found at the presence of nonviable R. solani. At the presence of nonviable R. solani, the maximum yield of trichodermin (144.55 mg/L) was significantly higher than the Control (67.8 mg/L), and the cultivation time to obtain the maximum yield of trichodermin decreased from 72 h to 60 h. No difference of trichodermin accumulation was observed by changing the concentration of nonviable R. solani from 0.1 to 1.6 g/L. It was observed that the optimum time for adding nonviable R. solani is immediately after inoculation. The diameter of T. brevicompactum mycelial globule after 72 h cultivation with nonviable R. solani elicitor was smaller than that of the Control. Xu-Ping Shentu, Wei-Ping Liu, Xiao-Huan Zhan, Xiao-Ping Yu, and Chuan-Xi Zhang Copyright © 2013 Xu-Ping Shentu et al. All rights reserved. Stakeholders’ Attitude to Genetically Modified Foods and Medicine Thu, 05 Dec 2013 13:01:29 +0000 Public acceptance of genetically modified (GM) foods has to be adequately addressed in order for their potential economic and social benefits to be realized. The objective of this paper is to assess the attitude of the Malaysian public toward GM foods (GM soybean and GM palm oil) and GM medicine (GM insulin). A survey was carried out using self-constructed multidimensional instrument measuring attitudes towards GM products. The respondents () were stratified according to stakeholders’ groups in the Klang Valley region. Results of the survey show that the overall attitude of the Malaysian stakeholders towards GM products was cautious. Although they acknowledged the presence of moderate perceived benefits associated with GM products surveyed and were moderately encouraging of them, they were also moderately concerned about the risks and moral aspects of the three GM products as well as moderately accepting the risks. Attitudes towards GM products among the stakeholders were found to vary not according to the type of all GM applications but rather depend on the intricate relationships between the attitudinal factors and the type of gene transfers involved. Analyses of variance showed significant differences in the six dimensions of attitude towards GM products across stakeholders’ groups. Latifah Amin, Jamaluddin Md Jahi, and Abd Rahim Md Nor Copyright © 2013 Latifah Amin et al. All rights reserved. Development of a pyrG Mutant of Aspergillus oryzae Strain S1 as a Host for the Production of Heterologous Proteins Sat, 30 Nov 2013 18:34:56 +0000 The ease with which auxotrophic strains and genes that complement them can be manipulated, as well as the stability of auxotrophic selection systems, are amongst the advantages of using auxotrophic markers to produce heterologous proteins. Most auxotrophic markers in Aspergillus oryzae originate from chemical or physical mutagenesis that may yield undesirable mutations along with the mutation of interest. An auxotrophic A. oryzae strain S1 was generated by deleting the orotidine-5′-monophosphate decarboxylase gene (pyrG) by targeted gene replacement. The uridine requirement of the resulting strain GR6 pyrGΔ0 was complemented by plasmids carrying a pyrG gene from either Aspergillus nidulans or A. oryzae. β-Galactosidase expression by strain GR6 pyrGΔ0 transformed with an A. niger plasmid encoding a heterologous β-galactosidase was at least 150 times more than that obtained with the untransformed strain. Targeted gene replacement is thus an efficient way of developing auxotrophic mutants in A. oryzae and the auxotrophic strain GR6 pyrGΔ0 facilitated the production of a heterologous protein in this fungus. Selina Oh Siew Ling, Reginald Storms, Yun Zheng, Mohd Rohaizad Mohd Rodzi, Nor Muhammad Mahadi, Rosli Md Illias, Abdul Munir Abdul Murad, and Farah Diba Abu Bakar Copyright © 2013 Selina Oh Siew Ling et al. All rights reserved. Stable Agrobacterium-Mediated Transformation of Maritime Pine Based on Kanamycin Selection Sun, 24 Nov 2013 14:09:55 +0000 An efficient transformation protocol based on kanamycin selection was developed for Agrobacterium-mediated transformation of maritime pine embryonal masses. The binary vector pBINUbiGUSint, which contained neomycin phosphotransferase II (nptII) as a selectable marker gene and β-glucuronidase (uidA) as a reporter gene, was used for transformation studies. Different factors, such as embryogenic line, bacterial strain, bacterial concentration, and coculture duration, were examined and optimized. For selection of transformants, 15 mgL−1 kanamycin was used. The highest transformation efficiency (11.4 events per gram of fresh mass) was achieved when a vigorously growing embryonal mass (embryogenic line L01) was cocultivated with Agrobacterium strain AGL1 at the optical density () of 0.3 for 72 h. Evidence of the stable transgene integration was obtained by polymerase chain reaction for the nptII and uidA genes and expression of the uidA gene. Maturation capacity of the transgenic lines was negatively affected by the transformation process. Induction of axillary shoots by preculturing the embryos with benzyladenine allowed overcoming the low maturation rates of some transformed lines. The transgenic embryos were germinated and the axillar shoots were rooted. Transgenic plants were transferred to potting substrate showing normal growth. José M. Alvarez and Ricardo J. Ordás Copyright © 2013 José M. Alvarez and Ricardo J. Ordás. All rights reserved. Characterization of a New Providencia sp. Strain X1 Producing Multiple Xylanases on Wheat Bran Wed, 20 Nov 2013 14:05:15 +0000 Providencia sp. strain X1 showing the highest xylanase activity among six bacterial isolates was isolated from saw-dust decomposing site. Strain X1 produced cellulase-free extracellular xylanase, which was higher in wheat bran medium than in xylan medium, when cultivated at pH 8.0 and 35°C. Zymogram analysis of crude preparation of enzymes obtained while growing on wheat bran and birchwood xylan revealed the presence of seven and two distinct xylanases with estimated molecular weight of 33; 35; 40; 48; 60; 75; and 95 kDa and 33 and 44 kDa, respectively. The crude xylanases were produced on wheat bran medium and showed optimum activity at pH 9.0 and 60°C. The thermotolerance studies showed activity retention of 100% and 85% at 40°C and 60°C after 30 min preincubation at pH 9.0. It was tolerant to lignin, ferulic acid, syringic acid, and guaiacol and retained 90% activity after ethanol treatment. The enzyme preparation was also tolerant to methanol and acetone and showed good activity retention in the presence of metal ions such as Fe2+, Mg2+, Zn2+, and Ca2+. The crude enzyme preparation was classified as endoxylanase based on the product pattern of xylan hydrolysis. Pretreatment of kraft pulp with crude xylanases for 3 h at 60°C led to a decrease in kappa number by 28.5%. The properties of present xylanases make them potentially useful for industrial applications. Abhay Raj, Sharad Kumar, Sudheer Kumar Singh, and Mahadeo Kumar Copyright © 2013 Abhay Raj et al. All rights reserved. Characterization of Thermo- and Detergent Stable Antigenic Glycosylated Cysteine Protease of Euphorbia nivulia Buch.-Ham. and Evaluation of Its Ecofriendly Applications Sun, 17 Nov 2013 14:31:34 +0000 An antigenic glycosylated cysteine protease has been purified from the latex of Euphorbia nivulia Buch.-Ham. It exhibits remarkable protease activity in the presence of metal ions, oxidizing agents, organic solvents, and detergents. This enzyme showed potential role in leather processing industry due to its dehairing activity for animal hide without hydrolyzing fibrous proteins, producing, by this way, a better quality product. The enzyme can also be used for silver recovering from X-ray plates. In addition, the stability (temperature and surfactants) and hydrolysis of blood stain data also revealed its application in detergent industries. Agriculturally, this protease finds application in biocontrol process against the infectious management of root knot nematode, Meloidogyne incognita. Biologically, it shows noticeable wound healing, haemostatic and antibacterial activity. Shamkant B. Badgujar and Raghunath T. Mahajan Copyright © 2013 Shamkant B. Badgujar and Raghunath T. Mahajan. All rights reserved. Characterization of Cellulolytic Bacterial Cultures Grown in Different Substrates Mon, 11 Nov 2013 14:23:52 +0000 Nine aerobic cellulolytic bacterial cultures were obtained from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Culture (DSMZ) and the American Type Culture Collection (ATCC). The objectives of this study were to characterize the cellulolytic bacteria and to determine the optimum moisture ratio required for solid state fermentation (SSF) of palm kernel cake (PKC). The bacteria cultures were grown on reconstituted nutrient broth, incubated at 30∘C and agitated at 200 rpm. Carboxymethyl cellulase, xylanase, and mannanase activities were determined using different substrates and after SSF of PKC. The SSF was conducted for 4 and 7 days with inoculum size of 10% (v/w) on different PKC concentration-to-moisture ratios: 1 : 0.2, 1 : 0.3, 1 : 0.4, and 1 : 0.5. Results showed that Bacillus amyloliquefaciens 1067 DSMZ, Bacillus megaterium 9885 ATCC, Paenibacillus curdlanolyticus 10248 DSMZ, and Paenibacillus polymyxa 842 ATCC produced higher enzyme activities as compared to other bacterial cultures grown on different substrates. The cultures mentioned above also produced higher enzyme activities when they were incubated under SSF using PKC as a substrate in different PKC-to-moisture ratios after 4 days of incubation, indicating that these cellulolytic bacteria can be used to degrade and improve the nutrient quality of PKC. Mohamed Idris Alshelmani, Teck Chwen Loh, Hooi Ling Foo, Wei Hong Lau, and Awis Qurni Sazili Copyright © 2013 Mohamed Idris Alshelmani et al. All rights reserved. Overexpression of luxS Cannot Increase Autoinducer-2 Production, Only Affect the Growth and Biofilm Formation in Streptococcus suis Thu, 07 Nov 2013 13:08:43 +0000 LuxS/AI-2 quorum sensing (QS) system involves the production of cell signaling molecules via luxS-based autoinducer-2 (AI-2). LuxS has been reported to plays critical roles in regulating various behaviors of bacteria. AI-2 is a byproduct of the catabolism of S-adenosylhomocysteine (SAH) performed by the LuxS and Pfs enzymes. In our previous study, the function of LuxS in AI-2 production was verified in Streptococcus suis (SS). Decreased levels of SS biofilm formation and host-cell adherence as well as an inability to produce AI-2 were observed in bacteria having a luxS mutant gene. In this study, the level of AI-2 activity exhibits a growth-phase dependence with a maximum in late exponential culture in SS. An SS strain that overexpressed luxS was constructed to comprehensively understand the function of AI-2. Overexpressed luxS was not able to increase the level of pfs expression and produce additional AI-2, and the bacteria were slower growing and produced only slightly more biofilm than the wild type. Thus, AI-2 production is not correlated with luxS transcription. luxS expression is constitutive, but the transcription of pfs is perhaps correlated with AI-2 production in SS. Yang Wang, Li Yi, Zhicheng Zhang, Hongjie Fan, Xiangchao Cheng, and Chengping Lu Copyright © 2013 Yang Wang et al. All rights reserved. Tyrosinase Inhibitory Effect and Antioxidative Activities of Fermented and Ethanol Extracts of Rhodiola rosea and Lonicera japonica Wed, 06 Nov 2013 09:12:33 +0000 This is the first study to investigate the biological activities of fermented extracts of Rhodiola rosea L. (Crassulaceae) and Lonicera japonica Thunb. (Caprifoliaceae). Alcaligenes piechaudii CC-ESB2 fermented and ethanol extracts of Rhodiola rosea and Lonicera japonica were prepared and the antioxidative activities of different concentrations of samples were evaluated using in vitro antioxidative assays. Tyrosinase inhibition was determined by using the dopachrome method with L-DOPA as substrate. The results demonstrated that inhibitory effects (ED50 values) on mushroom tyrosinase of fermented Rhodiola rosea, fermented Lonicera japonica, ethanol extract of Lonicera japonica, and ethanol extract of Rhodiola rosea were 0.78, 4.07, 6.93, and >10 mg/ml, respectively. The DPPH scavenging effects of fermented Rhodiola rosea (ED50 = 0.073 mg/ml) and fermented Lonicera japonica (ED50 = 0.207 mg/ml) were stronger than effects of their respective ethanol extracts. Furthermore, the scavenging effect increases with the presence of high content of total phenol. However, the superoxide scavenging effects of fermented Rhodiola rosea was less than effects of fermented Lonicera japonica. The results indicated that fermentation of Rhodiola rosea and Lonicera japonica can be considered as an effective biochemical process for application in food, drug, and cosmetics. Yuh-Shuen Chen, Hua-Chian Liou, and Chin-Feng Chan Copyright © 2013 Yuh-Shuen Chen et al. All rights reserved. Comparative Evaluation of Agroindustrial Byproducts for the Production of Alkaline Protease by Wild and Mutant Strains of Bacillus subtilis in Submerged and Solid State Fermentation Thu, 31 Oct 2013 15:33:20 +0000 The present study describes the screening of different agroindustrial byproducts for enhanced production of alkaline protease by a wild and EMS induced mutant strain of Bacillus subtilis IH-72EMS8. During submerged fermentation, different agro-industrial byproducts were tested which include defatted seed meals of rape, guar, sunflower, gluten, cotton, soybean, and gram. In addition to these meals, rice bran, wheat bran, and wheat flour were also evaluated for protease production. Of all the byproducts tested, soybean meal at a concentration of 20 g/L gave maximum production of the enzyme, that is, 5.74  ±  0.26 U/mL from wild and 11.28  ±  0.45 U/mL from mutant strain, during submerged fermentation. Different mesh sizes (coarse, medium, and fine) of the soybean meal were also evaluated, and a finely ground soybean meal (fine mesh) was found to be the best. In addition to the defatted seed meals, their alkali extracts were also tested for the production of alkaline protease by Bacillus subtilis, but these were proved nonsignificant for enhanced production of the enzyme. The production of the enzyme was also studied in solid state fermentation, and different agro-industrial byproducts were also evaluated for enzyme production. Wheat bran partially replaced with guar meal was found as the best substrate for maximum enzyme production under solid state fermentation conditions. Hamid Mukhtar and Ikramul Haq Copyright © 2013 Hamid Mukhtar and Ikramul Haq. All rights reserved. Large-Scale Plantlet Conversion and Ex Vitro Transplantation Efficiency of Siberian Ginseng by Bioreactor Culture Thu, 31 Oct 2013 09:18:58 +0000 To achieve large-scale low-cost ex vitro acclimatization of Siberian ginseng plants, heart- and torpedo-shaped secondary somatic embryos (SEs) induced from germinated SEs on agar medium were collected and then inoculated to 10-l bubble column bioreactor, respectively. For plantlet conversion, inoculation of torpedo-shaped secondary SEs was more effective than heart-shaped SEs. TS2 (culture of torpedo-shaped SEs in a bioreactor with a 2-week subculture interval) plantlets had a higher root number and leaf number and larger leaf area than did HS3 (culture of heart-shaped SEs in a bioreactor with a 3-week subculture interval) and HS2 (culture of heart-shaped SEs in a bioreactor with a 2-week subculture interval) plantlets. Of these converted plants, TS2 plantlets had higher survival rate (83.7%) and growth characteristics after transplantation in a simple shed covered with a 50% sunshade net only for 6 months. TS2 plantlets also showed significantly lower H2O2 content and significantly increased superoxide dismutase (SOD), glutathione peroxidase (GPX), and glutathione transferase (GST) expression levels as compared to HS2 plants when exposure to ex vitro conditions. Jingli Yang, Shicheng Zhao, Changyeon Yu, and Chenghao Li Copyright © 2013 Jingli Yang et al. All rights reserved. Dendrimers as Carriers for siRNA Delivery and Gene Silencing: A Review Tue, 29 Oct 2013 14:34:07 +0000 RNA interference (RNAi) was first literaturally reported in 1998 and has become rapidly a promising tool for therapeutic applications in gene therapy. In a typical RNAi process, small interfering RNAs (siRNA) are used to specifically downregulate the expression of the targeted gene, known as the term “gene silencing.” One key point for successful gene silencing is to employ a safe and efficient siRNA delivery system. In this context, dendrimers are emerging as potential nonviral vectors to deliver siRNA for RNAi purpose. Dendrimers have attracted intense interest since their emanating research in the 1980s and are extensively studied as efficient DNA delivery vectors in gene transfer applications, due to their unique features based on the well-defined and multivalent structures. Knowing that DNA and RNA possess a similar structure in terms of nucleic acid framework and the electronegative nature, one can also use the excellent DNA delivery properties of dendrimers to develop effective siRNA delivery systems. In this review, the development of dendrimer-based siRNA delivery vectors is summarized, focusing on the vector features (siRNA delivery efficiency, cytotoxicity, etc.) of different types of dendrimers and the related investigations on structure-activity relationship to promote safe and efficient siRNA delivery system. Jiangyu Wu, Weizhe Huang, and Ziying He Copyright © 2013 Jiangyu Wu et al. All rights reserved. Generation of Mature Nα-Terminal Acetylated Thymosin α1 by Cleavage of Recombinant Prothymosin α Mon, 28 Oct 2013 17:46:26 +0000 Nα-terminal acetylation of peptides plays an important biological role but is rarely observed in prokaryotes. Nα-terminal acetylated thymosin α1 (Tα1), a 28-amino-acid peptide, is an immune modifier that has been used in the clinic to treat hepatitis B and C virus (HBV/HCV) infections. We previously documented Nα-terminal acetylation of recombinant prothymosin α (ProTα) in E. coli. Here we present a method for production of Nα-acetylated Tα1 from recombinant ProTα. The recombinant ProTα was cleaved by human legumain expressed in Pichia pastoris to release Tα1 in vitro. The Nα-acetylated Tα1 peptide was subsequently purified by reverse phase and cation exchange chromatography. Mass spectrometry indicated that the molecular mass of recombinant Nα-acetylated Tα1 was 3108.79 in, which is identical to the mass of Nα-acetylated Tα1 produced by total chemical synthesis. This mass corresponded to the nonacetylated Tα1 mass with a 42 Da increment. The retention time of recombinant Nα-acetylated Tα1 and chemosynthetic Nα-acetylated Tα1 were both 15.4 min in RP-high performance liquid chromatography (HPLC). These data support the use of an E. coli expression system for the production of recombinant human Nα-acetylated Tα1 and also will provide the basis for the preparation of recombinant acetylated peptides in E. coli. Bo Liu, Xin Gong, Shaohong Chang, Peng Sun, and Jun Wu Copyright © 2013 Bo Liu et al. All rights reserved. A TaqMan Real-Time PCR Assay for Detection and Quantification of Sporisorium scitamineum in Sugarcane Mon, 21 Oct 2013 15:53:07 +0000 Sporisorium scitamineum is a fungal smut pathogen epidemic in sugarcane producing areas. Early detection and proper identification of the smut are an essential requirement in its management practice. In this study, we developed a TaqMan real-time PCR assay using specific primers (bEQ-F/bEQ-R) and a TaqMan probe (bEQ-P) which were designed based on the bE (b East mating type) gene (Genbank Accession no. U61290.1). This method was more sensitive (a detection limit of 10 ag pbE DNA and 0.8 ng sugarcane genomic DNA) than that of conventional PCR (10 fg and 100 ng, resp.). Reliability was demonstrated through the positive detection of samples collected from artificially inoculated sugarcane plantlets (FN40). This assay was capable of detecting the smut pathogen at the initial stage (12 h) of infection and suitable for inspection of sugarcane pathogen-free seed cane and seedlings. Furthermore, quantification of pathogen was verified in pathogen-challenged buds in different sugarcane genotypes, which suggested its feasibility for evaluation of smut resistance in different sugarcane genotypes. Taken together, this novel assay can be used as a diagnostic tool for sensitive, accurate, fast, and quantitative detection of the smut pathogen especially for asymptomatic seed cane or plants and evaluation of smut resistance of sugarcane genotypes. Yachun Su, Shanshan Wang, Jinlong Guo, Bantong Xue, Liping Xu, and Youxiong Que Copyright © 2013 Yachun Su et al. All rights reserved. Plant Biotechnology Thu, 10 Oct 2013 07:53:33 +0000 Khalid Mahmood Khawar, Selma Onarici, Cigdem Alev Ozel, Muhammad Aasim, Allah Bakhsh, and Abdul Qayyum Rao Copyright © 2013 Khalid Mahmood Khawar et al. All rights reserved. In Vitro Activation and Inhibition of Recombinant EGFR Tyrosine Kinase Expressed in Escherichia coli Wed, 25 Sep 2013 15:04:10 +0000 The present work concerns the heterologous expression of the intracellular domain harbouring the tyrosine kinase activity of the epidermal growth factor receptor (EGFR). Protein expression was improved thanks to the deletion of a 13-amino acid peptide of the juxtamembrane region (JM). The recombinant proteins were produced as a glutathione S-transferase (GST) fusion in Escherichia coli, and the solubilisation was performed by sarkosyl addition during extraction. The produced proteins spontaneously dimerize allowing the activation of the tyrosine kinase domain in the presence of ATP. The activity assay has revealed the autophosphorylation of EGFR proteins which was decreased in the presence of genistein. Our system could facilitate the screening of EGFR inhibitors without the need of adding an exogenous substrate. Jihene Elloumi-Mseddi, Karim Jellali, and Sami Aifa Copyright © 2013 Jihene Elloumi-Mseddi et al. All rights reserved. Elicitor-Induced Defense Responses in Solanum lycopersicum against Ralstonia solanacearum Wed, 25 Sep 2013 13:37:28 +0000 We investigated on important parameters of induced resistance in hydroponic tomato (Solanum lycopersicum) against Ralstonia solanacearum using the elicitors chitosan (CHT), salicylic acid (SA), and jasmonic acid (JA). The increase in total phenolic content of roots by the elicitors was significantly higher than control. Most pronounced increase in lignin synthesis was triggered by SA followed by CHT. At 24 h post-elicitation (hpe), the activity of phenylalanine ammonia lyase was 4.5 times higher than control elicited by CHT. The peroxidase activity was about 86 nkat/mg protein at 24 hpe in case of SA and 78 nkat/mg protein in case of CHT. The activity of polyphenol oxidase increased several folds by the elicitors. Cinnamyl alcohol dehydrogenase activity increased to the maximum at 48 hpe under the influence of CHT. The results indicate that the elicitors SA and CHT induced effective defense responses in tomato plants against R. solanacearum. This was evident from reduced vascular browning and wilting symptoms of tomato plants treated with SA and CHT and challenged subsequently with R. solanacearum. This reduced disease incidence in tomato by SA and CHT may be a result of cell wall strengthening through deposition of lignin and the coincident induction of defense enzymes. Sudhamoy Mandal, Itishree Kar, Arup K. Mukherjee, and Priyambada Acharya Copyright © 2013 Sudhamoy Mandal et al. All rights reserved. Optimization of Extraction Parameters by Using Response Surface Methodology, Purification, and Identification of Anthocyanin Pigments in Melastoma malabathricum Fruit Mon, 23 Sep 2013 12:02:58 +0000 Anthocyanins not just have various benefits in food industry but also have been used as natural colourants in cosmetic, coating products and as potential natural photosensitizers in solar cell. Thus, the main purpose of this study was to obtain information on the maximum yield of anthocyanin that can be recovered from Melastoma malabathricum fruit. Factors such as extraction temperature, extraction time, and solid to liquid ratio were identified to be significantly affecting anthocyanin extraction efficiency. By using three-level three-factor Box-Behnken design, the optimized conditions for anthocyanin extraction by acidified methanol () were temperature of 60°C, time of 86.82 min, and 0.5 : 35 (g/mL) solid to liquid ratio while the optimum extraction conditions by acidified ethanol () were temperature of 60°C, time of 120 min, and 0.5 : 23.06 (g/mL) solid to liquid ratio. The crude anthocyanin extract was further purified by using Amberlite XAD-7 and Sephadex LH-20 column chromatography. Identification of anthocyanins revealed the presence of cyanidin dihexoside, cyanidin hexoside, and delphinidin hexoside as the main anthocyanins in M. malabathricum fruit. Nordiyanah Anuar, Ahmad Faris Mohd Adnan, Naziz Saat, Norkasmani Aziz, and Rosna Mat Taha Copyright © 2013 Nordiyanah Anuar et al. All rights reserved. A Comparison of Ice Cold Water Pretreatment and α-Bromonaphthalene Cytogenetic Method for Identification of Papaver Species Wed, 11 Sep 2013 15:04:35 +0000 The plants belonging to many species in genus Papaver are very similar and have very small chromosomes that make identification very difficult. The study aimed to compare the effects of α-bromonaphtalene and ice cold water pretreatment to identify chromosomes of Papaver species collected from different areas of Iranian West Azerbaijan and Turkish Van, Agri and, Hakkari provinces. The seeds were germinated in Jacobson trays at 24°C under continuous light. Thereafter, roots from 1.5 cm long plantlets were pretreated with α bromonaphtalene for 15, 30, and 45 min or at 0°C in ice cold water for 24 h before fixing, hydrolyzation, and feulgen staining. The ice cold water pretreatment was more appropriate and easy to determine chromosomes. Seeds from seven samples did not germinate. Sixty samples out of the rest of 62 samples were identified as P. pseudo orientale, one sample was identified as P. bracteatum, and another as P. orientale. This is the first study that used ice cold water to determine the chromosomes in papaver species. It is hoped that it will also facilitate to determine chromosome number and identify other papver species. Amir Rezaei Osalou, Sheida Daneshvar Rouyandezagh, Behrouz Alizadeh, Celal Er, and Cafer Sirri Sevimay Copyright © 2013 Amir Rezaei Osalou et al. All rights reserved. Comparative Analyses of Response Surface Methodology and Artificial Neural Network on Medium Optimization for Tetraselmis sp. FTC209 Grown under Mixotrophic Condition Tue, 10 Sep 2013 14:28:36 +0000 Mixotrophic metabolism was evaluated as an option to augment the growth and lipid production of marine microalga Tetraselmis sp. FTC 209. In this study, a five-level three-factor central composite design (CCD) was implemented in order to enrich the W-30 algal growth medium. Response surface methodology (RSM) was employed to model the effect of three medium variables, that is, glucose (organic C source), NaNO3 (primary N source), and yeast extract (supplementary N, amino acids, and vitamins) on biomass concentration, , and lipid yield, /. RSM capability was also weighed against an artificial neural network (ANN) approach for predicting a composition that would result in maximum lipid productivity, . A quadratic regression from RSM and a Levenberg-Marquardt trained ANN network composed of 10 hidden neurons eventually produced comparable results, albeit ANN formulation was observed to yield higher values of response outputs. Finalized glucose (24.05 g/L), NaNO3 (4.70 g/L), and yeast extract (0.93 g/L) concentration, affected an increase of to 12.38 g/L and lipid a accumulation of 195.77 mg/g dcw. This contributed to a lipid productivity of 173.11 mg/L per day in the course of two-week cultivation. Mohd Shamzi Mohamed, Joo Shun Tan, Rosfarizan Mohamad, Mohd Noriznan Mokhtar, and Arbakariya B. Ariff Copyright © 2013 Mohd Shamzi Mohamed et al. All rights reserved. Induction and Analysis of the Alkaloid Mitragynine Content of a Mitragyna speciosa Suspension Culture System upon Elicitation and Precursor Feeding Mon, 26 Aug 2013 10:27:25 +0000 This study aimed to determine the effects of different concentrations and combinations of the phytohormones 2,4-dichlorophenoxy acetic acid (2,4-D), kinetin, 6-benzylaminopurine (BAP), and 1-naphthaleneacetic acid (NAA) on callus induction and to demonstrate the role of elicitors and exogenous precursors on the production of mitragynine in a Mitragyna speciosa suspension culture. The best callus induction was achieved from petiole explants cultured on WPM that was supplemented with 4 mg L−1 2, 4-D (70.83%). Calli were transferred to liquid media and agitated on rotary shakers to establish Mitragyna speciosa cell suspension cultures. The optimum settled cell volume was achieved in the presence of WPM that contained 3 mg L−1 2,4-D and 3% sucrose ( mL). The treatment of cultures with different concentrations of yeast extract and salicylic acid for different inoculation periods revealed that the highest mitragynine content as determined by HPLC was achieved from the culture treated with 250 mg L−1 yeast extract ( mg L−1) that was harvested on day 6 of culturing; salicylic acid showed low mitragynine content in all concentrations used. Tryptophan and loganin were used as exogenous precursors; the highest level of mitragynine production was achieved in cultures treated with 3 μM tryptophan and harvested at 6 days ( mg L−1). Nor Nahazima Mohamad Zuldin, Ikram Md. Said, Normah Mohd Noor, Zamri Zainal, Chew Jin Kiat, and Ismanizan Ismail Copyright © 2013 Nor Nahazima Mohamad Zuldin et al. All rights reserved. Annotation of Differentially Expressed Genes in the Somatic Embryogenesis of Musa and Their Location in the Banana Genome Wed, 21 Aug 2013 13:27:11 +0000 Analysis of cDNA-AFLP was used to study the genes expressed in zygotic and somatic embryogenesis of Musa acuminata Colla ssp. malaccensis, and a comparison was made between their differential transcribed fragments (TDFs) and the sequenced genome of the double haploid- (DH-) Pahang of the malaccensis subspecies that is available in the network. A total of 253 transcript-derived fragments (TDFs) were detected with apparent size of 100–4000 bp using 5 pairs of AFLP primers, of which 21 were differentially expressed during the different stages of banana embryogenesis; 15 of the sequences have matched DH-Pahang chromosomes, with 7 of them being homologous to gene sequences encoding either known or putative protein domains of higher plants. Four TDF sequences were located in all Musa chromosomes, while the rest were located in one or two chromosomes. Their putative individual function is briefly reviewed based on published information, and the potential roles of these genes in embryo development are discussed. Thus the availability of the genome of Musa and the information of TDFs sequences presented here opens new possibilities for an in-depth study of the molecular and biochemical research of zygotic and somatic embryogenesis of Musa. Josefina Ines Maldonado-Borges, José Roberto Ku-Cauich, and Rosa Maria Escobedo-GraciaMedrano Copyright © 2013 Josefina Ines Maldonado-Borges et al. All rights reserved. Genetic Diversity Analysis of Sugarcane Parents in Chinese Breeding Programmes Using gSSR Markers Wed, 07 Aug 2013 10:51:19 +0000 Sugarcane is the most important sugar and bioenergy crop in the world. The selection and combination of parents for crossing rely on an understanding of their genetic structures and molecular diversity. In the present study, 115 sugarcane genotypes used for parental crossing were genotyped based on five genomic simple sequence repeat marker (gSSR) loci and 88 polymorphic alleles of loci (100%) as detected by capillary electrophoresis. The values of genetic diversity parameters across the populations indicate that the genetic variation intrapopulation (90.5%) was much larger than that of interpopulation (9.5%). Cluster analysis revealed that there were three groups termed as groups I, II, and III within the 115 genotypes. The genotypes released by each breeding programme showed closer genetic relationships, except the YC series released by Hainan sugarcane breeding station. Using principle component analysis (PCA), the first and second principal components accounted for a cumulative 76% of the total variances, in which 43% were for common parents and 33% were for new parents, respectively. The knowledge obtained in this study should be useful to future breeding programs for increasing genetic diversity of sugarcane varieties and cultivars to meet the demand of sugarcane cultivation for sugar and bioenergy use. Qian You, Liping Xu, Yifeng Zheng, and Youxiong Que Copyright © 2013 Qian You et al. All rights reserved. Genetic Transformation in Citrus Thu, 25 Jul 2013 14:13:58 +0000 Citrus is one of the world’s important fruit crops. Recently, citrus molecular genetics and biotechnology work have been accelerated in the world. Genetic transformation, a biotechnological tool, allows the release of improved cultivars with desirable characteristics in a shorter period of time and therefore may be useful in citrus breeding programs. Citrus transformation has now been achieved in a number of laboratories by various methods. Agrobacterium tumefaciens is used mainly in citrus transformation studies. Particle bombardment, electroporation, A. rhizogenes, and a new method called RNA interference are used in citrus transformation studies in addition to A. tumefaciens. In this review, we illustrate how different gene transformation methods can be employed in different citrus species. Dicle Donmez, Ozhan Simsek, Tolga Izgu, Yildiz Aka Kacar, and Yesim Yalcin Mendi Copyright © 2013 Dicle Donmez et al. All rights reserved. Micropropagation of Origanum acutidens (HAND.-MAZZ.) IETSWAART Using Stem Node Explants Wed, 24 Jul 2013 14:10:29 +0000 Origanum acutidens (HAND.-MAZZ.) IETSWAART is a promising ornamental plant that can be widely used in landscape management. It is endemic to Eastern Anatolian region of Turkey. Tissue culture has not been used to micropropagate it. The study reports stem node explants from one-week-old seedlings of the plant for successful micropropagation. The stem nodes were cultured on MS medium containing 0.6, 1.2, 1.8, and 2.4 mg/L BAP with 0.2 mg/L NAA. Visible effects of culture media on shoot proliferation were recorded. Shoot regeneration rate was maximum on MS medium containing 1.80 mg/L BAP-0.2 mg/L NAA. The micropropagated shoots were rooted on MS medium containing 0.2 mg/L NAA. All microrooted plantlets survived during acclimatisation on peat moss. It was concluded that O. acutidens can be successfully micropropagated under in vitro conditions. Mehmet Ugur Yildirim Copyright © 2013 Mehmet Ugur Yildirim. All rights reserved. The Influence of 1-Triacontanol on the Growth, Flowering, and Quality of Potted Bougainvillea Plants (Bougainvillea glabra var. “Elizabeth Angus”) under Natural Conditions Wed, 17 Jul 2013 15:33:11 +0000 Selected physiological and biochemical parameters were monitored at the vegetative and reproductive growth stages in potted Bougainvillea plants treated with five different concentrations of TRIA. Advanced flowering, flower bud number, and blooming rate increased significantly with 0.5 and 1.0 mg/L TRIA treatments. Similarly, photosynthetic rate, pigment content, quantum yield, and stomatal conductance increased significantly with 2.5, 1.0, and 5.0 mg/L TRIA treatments. Higher levels of N, P, and K, as well as increased total soluble solids (TSS) and higher sugar and protein contents, were recorded in treated plants. Furthermore, 46% more flowers, a 1.5-fold increase in bract weight, increased longevity, and 40% less leaf abscission were recorded following 2.5 mg/L TRIA treatment. Phenol and flavonoid contents, sucrose phosphate synthase (SPS), and antioxidant activities were also markedly increased with 2.5 and 1.0 mg/L TRIA treatments. However, ethylene production was significantly lower in the treated plants. Positive correlations were observed between leaf TSS and flowering time and flower number, between leaf sugar content and bract weight, and between net photosynthesis and bract growth and dry matter production. It can be concluded that the foliar spray of TRIA stimulates growth, enhances flowering, and improves the quality of potted Bougainvillea plants. Mohammad Moneruzzaman Khandaker, Golam Faruq, M. Motior Rahman, M. Sofian-Azirun, and Amru Nasrulhaq Boyce Copyright © 2013 Mohammad Moneruzzaman Khandaker et al. All rights reserved. Use of Tissue Culture Techniques for Producing Virus-Free Plant in Garlic and Their Identification through Real-Time PCR Sun, 07 Jul 2013 10:53:38 +0000 This study was performed for comparison of meristem culture technique with shoot tip culture technique for obtaining virus-free plant, comparison of micropropagation success of two different nutrient media, and determination of effectiveness of real-time PCR assay for the detection of viruses. Two different garlic species (Allium sativum and Allium tuncelianum) and two different nutrient media were used in this experiment. Results showed that Medium 2 was more successful compared to Medium 1 for both A. tuncelianum and A. sativum (Kastamonu garlic clone). In vitro plants obtained via meristem and shoot tip cultures were tested for determination of onion yellow dwarf virus (OYDV) and leek yellow stripe virus (LYSV) through real-time PCR assay. In garlic plants propagated via meristem culture, we could not detect any virus. OYDV and LYSV viruses were detected in plants obtained via shoot tip culture. OYDV virus was observed in amount of 80% and 73% of tested plants for A. tuncelianum and A. sativum, respectively. LYSV virus was found in amount of 67% of tested plants of A. tuncelianum and in amount of 87% of tested plants of A. sativum in this study. Hatıra Taşkın, Gökhan Baktemur, Mehmet Kurul, and Saadet Büyükalaca Copyright © 2013 Hatıra Taşkın et al. All rights reserved. Adventitious Shoot Regeneration from Leaf Explant of Dwarf Hygro (Hygrophila polysperma (Roxb.) T. Anderson) Mon, 17 Jun 2013 18:43:00 +0000 Dwarf hygro (Hygrophila polysperma) is an ornamental aquatic plant that changes its leaf colours to pinkish in high light. It is listed as a medicinal plant in medicinal plant lists of Indian states of West Bengal and Karnataka. It is also used as a screening tool for toxicities and a bioindicator to detect and control algae. The study reported in vitro adventitious shoot regeneration from leaf explants cultured on MS medium containing 0.10–1.60 mg/L Kin/TDZ with or without 0.10 mg/L IBA and 500 mg/L Amoklavin to eradicate endogenic bacterial contamination. Direct adventitious shoot regeneration started within one week from both culture mediums followed by late callus induction which was more prominent on TDZ containing media compared to Kin containing media. Addition of 0.10 mg/L IBA with both Kin and TDZ increased shoot regeneration frequency, mean number of shoots per explant, and mean shoot length. Maximum number of 16.33 and 20.55 shoots per explant was obtained on MS medium containing  mg/L Kin-IBA and  mg/L TDZ-IBA, respectively. Regenerated shoots were rooted on MS medium containing 0.20–1.00 mg/L IBA followed by successfull acclimatization in aquariums. Regenerated plantlets were also tested in jars containing distilled water that showed the pH 6–9 for the best plant growth and development. Mehmet Karataş, Muhammad Aasim, Ayşegül Çınar, and Muhammet Dogan Copyright © 2013 Mehmet Karataş et al. All rights reserved. Protein Profiling and Histone Deacetylation Activities in Somaclonal Variants of Oil Palm (Elaeis guineensis Jacq.) Mon, 03 Jun 2013 16:44:56 +0000 Mantled fruits as a result of somaclonal variation are often observed from the oil palm plantlets regenerated via tissue culture. The mantling of fruits with finger-like and thick outer coating phenotypes significantly reduces the seed size and oil content, posing a threat to oil palm planters, and may jeopardize the economic growth of countries that depend particularly on oil palm plantation. The molecular aspects of the occurrence of somaclonal variations are yet to be known, possibly due to gene repression such as DNA methylation, histone methylation and histone deacetylation. Histone deacetylases (HDACs), involved in eukaryotic gene regulation by catalyzing the acetyl groups are removal from lysine residues on histone, hence transcriptionally repress gene expression. This paper described the total protein polymorphism profiles of somaclonal variants of oil palm and the effects of histone deacetylation on this phenomenon. Parallel to the different phenotypes, the protein polymorphism profiles of the mantled samples (leaves, fruits, and florets) and the phenotypically normal samples were proven to be different. Higher HDAC activity was found in mantled leaf samples than in the phenotypically normal leaf samples, leading to a preliminary conclusion that histone deacetylation suppressed gene expression and contributed to the development of somaclonal variants. Jamilah Syafawati Yaacob, Hwei-San Loh, and Rosna Mat Taha Copyright © 2013 Jamilah Syafawati Yaacob et al. All rights reserved. Effects of IAA, IBA, NAA, and GA3 on Rooting and Morphological Features of Melissa officinalis L. Stem Cuttings Thu, 30 May 2013 12:04:24 +0000 This study analyzed the potential of producing Melissa officinalis L. using stem cuttings. Four different hormones (IAA, IBA, NAA, and GA3) were applied to the cuttings, with and without buds, in two doses (1000 mg/L and 5000 mg/L), and after 60 days, 10 morphological characteristics of newly generated plants were detected, and a statistical analysis was carried out. The results of the study show that the cuttings with at least one bud must be used in order to produce M. officinalis using stem cuttings. Even though the auxin group hormones (IAA, IBA, and NAA) do not have an apparent effect on rooting percentage, these hormones were detected to affect the morphological characteristics of the newly generated plants, especially root generation. GA3 application has a considerable effect on stem height. Hakan Sevik and Kerim Guney Copyright © 2013 Hakan Sevik and Kerim Guney. All rights reserved. High Efficiency Secondary Somatic Embryogenesis in Hovenia dulcis Thunb. through Solid and Liquid Cultures Wed, 29 May 2013 18:10:34 +0000 Embryogenic callus was obtained from mature seed explants on medium supplemented with 2,4-dichlorophenoxyacetic acid. Primary somatic embryos (SEs) can only develop into abnormal plants. Well-developed SEs could be obtained through secondary somatic embryogenesis both in solid and liquid cultures. Temperature strongly affected induction frequency of secondary embryogenesis. Relatively high temperature () and germinated SEs explants were effective for induction of secondary somatic embryos, and low temperature () was more suitable for further embryo development, plantlet conversion, and transplant survival. Somatic embryos formed on agar medium had larger cotyledons than those of embryos formed in liquid medium. Supplementing 0.1 mg  6-benzyladenine (BA) was effective for plant conversion; the rate of plant conversion was 43.3% in somatic embryos from solid culture and 36.5% in embryos from liquid culture. In vitro plants were successfully acclimatized in the greenhouse. The protocol established in this study will be helpful for large-scale vegetative propagation of this medicinal tree. Jingli Yang, Songquan Wu, and Chenghao Li Copyright © 2013 Jingli Yang et al. All rights reserved. Comparison of Different Methods for Separation of Haploid Embryo Induced through Irradiated Pollen and Their Economic Analysis in Melon (Cucumis melo var. inodorus) Wed, 29 May 2013 18:10:02 +0000 Irradiated pollen technique is the most successful haploidization technique within Cucurbitaceae. After harvesting of fruits pollinated with irradiated pollen, classical method called as “inspecting the seeds one by one” is used to find haploid embryos in the seeds. In this study, different methods were used to extract the embryos more easily, quickly, economically, and effectively. “Inspecting the seeds one by one” was used as control treatment. Other four methods tested were “sowing seeds direct nutrient media,” “inspecting seeds in the light source,” “floating seeds on liquid media,” and “floating seeds on liquid media after surface sterilization.” Y2 and Y3 melon genotypes selected from the third backcross population of Yuva were used as plant material. Results of this study show that there is no statistically significant difference among methods “inspecting the seeds one by one,” “sowing seeds direct CP nutrient media,” and “inspecting seeds in the light source,” although the average number of embryos per fruit is slightly different. No embryo production was obtained from liquid culture because of infection. When considered together with labor costs and time required for embryo rescue, the best methods were “sowing seeds directly in the CP nutrient media“ and ”inspecting seeds in the light source.” Gökhan Baktemur, Hatıra Taşkın, and Saadet Büyükalaca Copyright © 2013 Gökhan Baktemur et al. All rights reserved. Comparative Studies on Cellular Behaviour of Carnation (Dianthus caryophyllus Linn. cv. Grenadin) Grown In Vivo and In Vitro for Early Detection of Somaclonal Variation Wed, 22 May 2013 15:55:52 +0000 The present study deals with the cytological investigations on the meristematic root cells of carnation (Dianthus caryophyllus Linn.) grown in vivo and in vitro. Cellular parameters including the mitotic index (MI), chromosome count, ploidy level (nuclear DNA content), mean cell and nuclear areas, and cell doubling time (Cdt) were determined from the 2 mm root tip segments of this species. The MI value decreased when cells were transferred from in vivo to in vitro conditions, perhaps due to early adaptations of the cells to the in vitro environment. The mean chromosome number was generally stable () throughout the 6-month culture period, indicating no occurrence of early somaclonal variation. Following the transfer to the in vitro environment, a significant increase was recorded for mean cell and nuclear areas, from 26.59 ± 0.09 μm2 to 35.66 ± 0.10 μm2 and 142.90 ± 0.59 μm2 to 165.05 ± 0.58 μm2, respectively. However, the mean cell and nuclear areas of in vitro grown D. caryophyllus were unstable and fluctuated throughout the tissue culture period, possibly due to organogenesis or rhizogenesis. Ploidy level analysis revealed that D. caryophyllus root cells contained high percentage of polyploid cells when grown in vivo and maintained high throughout the 6-month culture period. Jamilah Syafawati Yaacob, Rosna Mat Taha, and Arash Khorasani Esmaeili Copyright © 2013 Jamilah Syafawati Yaacob et al. All rights reserved. Sponge-Like: A New Protocol for Preparing Bacterial Ghosts Mon, 18 Mar 2013 09:42:24 +0000 Bacterial Ghosts (BGs) received an increasing interest in the recent years for their promising medicinal and pharmaceutical applications. In this study, for the first time we introduce a new protocol for BGs production. E. coli BL21 (DE3) pLysS (Promega) was used as a model to establish a general protocol for BGs preparation. The protocol is based on using active chemical compounds in concentrations less than the Minimum Inhibition Concentration (MIC). Those chemical compounds are SDS, NaOH, and H2O2. Plackett-Burman experimental design was used to map the best conditions for BGs production. Normal and electronic microscopes were used to evaluate the BGs quality (BGQ). Spectrophotometer was used to evaluate the amount of the released protein and DNA. Agarose gel electrophoresis was used to determine the existence of any residue of DNA after each BGs preparation. Viable cells, which existed after running this protocol, were subjected to lysis by inducing the lysozyme gene carried on pLysS plasmid. This protocol is able to produce BGs that can be used in different biotechnological applications. Amro A. Amara, Mounir M. Salem-Bekhit, and Fars K. Alanazi Copyright © 2013 Amro A. Amara et al. All rights reserved. Germination and Plantlet Regeneration of Encapsulated Microshoots of Aromatic Rice (Oryza sativa L. Cv. MRQ 74) Thu, 02 Aug 2012 09:27:34 +0000 Plant tissues such as somatic embryos, apical shoot tips, axillary shoot buds, embryogenic calli, and protocom-like bodies are potential micropropagules that have been considered for creating synthetic seeds. In the present study, 3–5 mm microshoots of Oryza sativa L. Cv. MRQ 74 were used as explant sources for obtaining synthetic seeds. Microshoots were induced from stem explants on Murashige and Skoog (MS) medium supplemented with 1.5 mg/L benzylaminopurine (BAP). They were encapsulated in 3% (w/v) sodium alginate, 3% sucrose, 0.1 mg/L BAP, and 0.1 mg/L α-Naphthalene acetic acid (NAA). Germination and plantlet regeneration of the encapsulated seeds were tested by culturing them on various germination media. The effect of storage period (15–30 days) was also investigated. The maximum germination and plantlet regeneration (100.0%) were recorded on MS media containing 3% sucrose and 0.8% agar with and without 0.1 mg/L BAP. However, a low germination rate (6.67%) was obtained using top soil as a sowing substrate. The germination rate of the encapsulated microshoots decreased from 93.33% to 3.33% after 30 days of storage at 4°C in the dark. Therefore, further research is being done to improve the germination rate of the synthetic seeds. Rosna Mat Taha, Azani Saleh, Noraini Mahmad, Nor Azlina Hasbullah, and Sadegh Mohajer Copyright © 2012 Rosna Mat Taha et al. All rights reserved. FAD2-DGAT2 Genes Coexpressed in Endophytic Aspergillus fumigatus Derived from Tung Oilseeds Tue, 31 Jul 2012 10:52:59 +0000 Recent efforts to genetically engineer plants that contain fatty acid desaturases to produce valuable fatty acids have made only modest progress. Diacylglycerol acyltransferase 2 (DGAT2), which catalyzes the final step in triacylglycerol (TAG) assembly, might potentially regulate the biosynthesis of desired fatty acids in TAGs. To study the effects of tung tree (Vernicia fordii) vfDGAT2 in channeling the desired fatty acids into TAG, vfDGAT2 combined with the tung tree fatty acid desaturase-2 (vfFAD2) gene was co-introduced into Aspergillus fumigatus, an endophytic fungus isolated from healthy tung oilseed. Two transformants coexpressing vfFAD2 and vfDGAT2 showed a more than 6-fold increase in linoleic acid production compared to the original A. fumigatus strain, while a nearly 2-fold increase was found in the transformant expressing only vfFAD2. Our data suggest that vfDGAT2 plays a pivotal role in promoting linoleic acid accumulation in TAGs. This holds great promise for further genetic engineering aimed at producing valuable fatty acids. Yi-Cun Chen, Yang-Dong Wang, Qin-Qin Cui, and Zhi-Yong Zhan Copyright © 2012 Yi-Cun Chen et al. All rights reserved. Bioremediation of Parboiled Rice Effluent Supplemented with Biodiesel-Derived Glycerol Using Pichia pastoris X-33 Tue, 31 Jul 2012 10:35:27 +0000 This paper describes the use of Pichia pastoris X-33 as a bioremediator to reduce the chemical oxygen demand (COD), total Kjeldahl nitrogen (TKN), and phosphorus (P-PO43−), after culture in parboiled rice effluent supplemented with p.a. glycerol or a glycerol by-product of the biodiesel industry. The greatest reduction in the COD (55%), TKN (45%), and P-PO43− (52%) of the effluent was observed in cultures of P. pastoris X-33 supplemented with 15 g ·L−1 of biodiesel-derived glycerol. Furthermore, the overall biomass yield was 2.1 g ·L−1. These data suggest that biodiesel-derived glycerol is an efficient carbon source for the bioremediation of parboiled rice effluent and biomass production. Diego Gil de los Santos, Carlos Gil Turnes, and Fabricio Rochedo Conceição Copyright © 2012 Diego Gil de los Santos et al. All rights reserved. Postharvest Analysis of Lowland Transgenic Tomato Fruits Harboring hpRNAi-ACO1 Construct Tue, 31 Jul 2012 10:32:00 +0000 The plant hormone, ethylene, is an important regulator which involved in regulating fruit ripening and flower senescence. In this study, RNA interference (RNAi) technology was employed to silence the genes involved in ethylene biosynthetic pathway. This was achieved by blocking the expression of specific gene encoding the ACC oxidase. Initially, cDNA corresponding to ACO1 of lowland tomato cultivar (MT1), which has high identity with ACO1 of Solanum lycopersicum in GenBank, was cloned through RT-PCR. Using a partial coding region of ACO1, one hpRNAi transformation vector was constructed and expressed ectopically under the 35S promoter. Results showed that transgenic lines harboring the hpRNA-ACO1 construct had lower ethylene production and a longer shelf life of 32 days as compared to 10 days for wild-type fruits. Changes in cell wall degrading enzyme activities were also investigated in cases where the transgenic fruits exhibited reduced rates of firmness loss, which can be associated with a decrease in pectin methylesterase (PME) and polygalacturonase (PG) activities. However, no significant change was detected in both transgenic and wild-type fruits in terms of β-galactosidase (β-Gal) activity and levels of total soluble solid, titratable acid and ascorbic acid. Bita Behboodian, Zainon Mohd Ali, Ismanizan Ismail, and Zamri Zainal Copyright © 2012 Bita Behboodian et al. All rights reserved. Distinct Mechanisms Regulating Gene Expression Coexist within the Fermentative Pathways in Chlamydomonas reinhardtii Mon, 18 Jun 2012 10:19:16 +0000 Under dark anoxia, the unicellular green algae Chlamydomonas reinhardtii may produce hydrogen by means of its hydrogenase enzymes, in particular HYD1, using reductants derived from the degradation of intercellular carbon stores. Other enzymes belonging to the fermentative pathways compete for the same reductants. A complete understanding of the mechanisms determining the activation of one pathway rather than another will help us engineer Chlamydomonas for fermentative metabolite production, including hydrogen. We examined the expression pattern of the fermentative genes PDC3, LDH1, ADH2, PFL1, and PFR1 in response to day-night cycles, continuous light, continuous darkness, and low or high oxygen availability, which are all conditions that vary on a regular basis in Chlamydomonas' natural environment. We found that all genes except PFL1 show daily fluctuations in expression, and that PFR1 differentiated itself from the others in that it is clearly responsive to low oxygen, where as PDC3, LDH1, and ADH2 are primarily under diurnal regulation. Our results provide evidence that there exist at least three different regulatory mechanisms within the fermentative pathways and suggest that the fermentative pathways are not redundant but rather that availability of a variety of pathways allows for a differential metabolic response to different environmental conditions. Larisa Angela Swirsky Whitney, Giacomo Novi, Pierdomenico Perata, and Elena Loreti Copyright © 2012 Larisa Angela Swirsky Whitney et al. All rights reserved. Genetic Diversity of Arabica Coffee (Coffea arabica L.) in Nicaragua as Estimated by Simple Sequence Repeat Markers Mon, 04 Jun 2012 16:02:40 +0000 Coffea arabica L. (arabica coffee), the only tetraploid species in the genus Coffea, represents the majority of the world’s coffee production and has a significant contribution to Nicaragua’s economy. The present paper was conducted to determine the genetic diversity of arabica coffee in Nicaragua for its conservation and breeding values. Twenty-six populations that represent eight varieties in Nicaragua were investigated using simple sequence repeat (SSR) markers. A total of 24 alleles were obtained from the 12 loci investigated across 260 individual plants. The total Nei’s gene diversity (𝐻𝑇) and the within-population gene diversity (𝐻𝑆) were 0.35 and 0.29, respectively, which is comparable with that previously reported from other countries and regions. Among the varieties, the highest diversity was recorded in the variety Catimor. Analysis of variance (AMOVA) revealed that about 87% of the total genetic variation was found within populations and the remaining 13% differentiate the populations (𝐹𝑆𝑇=0.13; 𝑃<0.001). The variation among the varieties was also significant. The genetic variation in Nicaraguan coffee is significant enough to be used in the breeding programs, and most of this variation can be conserved through ex situ conservation of a low number of populations from each variety. Mulatu Geleta, Isabel Herrera, Arnulfo Monzón, and Tomas Bryngelsson Copyright © 2012 Mulatu Geleta et al. All rights reserved. Effect of ADH II Deficiency on the Intracellular Redox Homeostasis in Zymomonas mobilis Thu, 03 May 2012 08:39:32 +0000 Mutant strain of the facultatively anaerobic, ethanol-producing bacterium Zymomonas mobilis, deficient in the Fe-containing alcohol dehydrogenase isoenzyme (ADH II), showed impaired homeostasis of the intracellular NAD(P)H during transition from anaerobic to aerobic conditions, and also in steady-state continuous cultures at various oxygen supplies. At the same time, ADH II deficiency in aerobically grown cells was accompanied by a threefold increase of catalase activity and by about 50% increase of hydrogen peroxide excretion. It is concluded that ADH II under aerobic conditions functions to maintain intracellular redox homeostasis and to protect the cells from endogenous hydrogen peroxide. Nina Galinina, Zane Lasa, Inese Strazdina, Reinis Rutkis, and Uldis Kalnenieks Copyright © 2012 Nina Galinina et al. All rights reserved. Animal Bones Char Solubilization by Gel-Entrapped Yarrowia lipolytica on Glycerol-Based Media Thu, 03 May 2012 08:34:42 +0000 Citric acid was produced with free and k-carrageenan-entrapped cells of the yeast Yarrowia lipolytica in single and repeated batch-shake-flask fermentations on glycerol-based media. Simultaneous solubilization of hydroxyapatite of animal bone origin (HABO) was tested in all experiments. The highest citric acid production by free yeast cells of 20.4 g/L and 18.7 g/L was reached after 96 h of fermentation in the absence and presence of 3 g/L HABO, respectively. The maximum values for the same parameter achieved by gel-entrapped cells in conditions of single batch and repeated-batch fermentation processes were 18.7 g/L and 28.1 g/L registered after 96 h and the 3d batch cycle, respectively. The highest citric acid productivity of 0.58 g L−1 h−1 was obtained with immobilized cells in repeated batch mode of fermentation when the added hydroxyapatite of 3 g/L was solubilized to 399 mg/L whereas the maximum efficiency of 89.0% was obtained with 1 g/L of HABO. Maria Vassileva, Bettina Eichler-Lobermann, Antonia Reyes, and Nikolay Vassilev Copyright © 2012 Maria Vassileva et al. All rights reserved. Production Response and Digestive Enzymatic Activity of the Pacific White Shrimp Litopenaeus vannamei (Boone, 1931) Intensively Pregrown in Microbial Heterotrophic and Autotrophic-Based Systems Wed, 02 May 2012 11:57:48 +0000 Shrimp postlarvae were reared into different microcosm systems without water exchange; a traditional system based on simple fertilization to improve microalgae concentration (control), an autotrophic system (AS) based on the promotion of biofloc and biofilm by the addition of fertilizer and artificial substrates and a heterotrophic system (HS) based on the promotion of heterotrophic bacteria by the addition of nitrogenous and carbonaceous sources and artificial substrates. Better growth performance and survival were registered in shrimp from the AS and HS compared to the control. Feed conversion ratios were below 0.7 for all treatments, but AS and HS were significantly lower than the control. Regarding digestive performance, no significant differences were observed for trypsin, amylase and lipase activities among AS and control shrimp; however, shrimp from HS showed a higher trypsin and amylase activities, suggesting a higher digestive activity caused by the presence of microbial bioflocs. The presence of biofilm and bioflocs composed by either autotrophic or heterotrophic organisms in combination with formulated feed improved the growth performance and survival of shrimp. Apparently, such combination fits the nutritional requirements of shrimp. Manuel J. Becerra-Dórame, Marcel Martínez-Porchas, Luis R. Martínez-Córdova, Martha E. Rivas-Vega, José A. Lopez-Elias, and Marco A. Porchas-Cornejo Copyright © 2012 Manuel J. Becerra-Dórame et al. All rights reserved. Use of Enzymatic Bio-Fenton as a New Approach in Decolorization of Malachite Green Wed, 02 May 2012 11:43:30 +0000 An enzymatic reaction using glucose oxidase was applied for in situ production of hydrogen peroxide for use in simultaneously Fenton's reaction in decolorization of malachite green. It was found that decolorization rate increased by increasing of glucose concentration from 0.2 g/L to 1.5 g/L. Decolorization rate showed different behaviors versus temperature changes. Initial rate of decolorization process was increased by increasing of temperature; after 30 minutes, especially at temperatures above 30°C, the decolorization rate was gradually reduced. The pH value in the reaction media was decreased from natural to about pH=3 which had synergic effect on the Fenton process by stabilizing of Fe2+ ions. Afzal Karimi, Mostafa Aghbolaghy, Alireza Khataee, and Shabnam Shoa Bargh Copyright © 2012 Afzal Karimi et al. All rights reserved. Removal of Triphenylmethane Dyes by Bacterial Consortium Tue, 01 May 2012 15:48:48 +0000 A new consortium of four bacterial isolates (Agrobacterium radiobacter; Bacillus spp.; Sphingomonas paucimobilis, and Aeromonas hydrophila)-(CM-4) was used to degrade and to decolorize triphenylmethane dyes. All bacteria were isolated from activated sludge extracted from a wastewater treatment station of a dyeing industry plant. Individual bacterial isolates exhibited a remarkable color-removal capability against crystal violet (50 mg/L) and malachite green (50 mg/L) dyes within 24 h. Interestingly, the microbial consortium CM-4 shows a high decolorizing percentage for crystal violet and malachite green, respectively, 91% and 99% within 2 h. The rate of chemical oxygen demand (COD) removal increases after 24 h, reaching 61.5% and 84.2% for crystal violet and malachite green, respectively. UV-Visible absorption spectra, FTIR analysis and the inspection of bacterial cells growth indicated that color removal by the CM-4 was due to biodegradation. Evaluation of mutagenicity by using Salmonella typhimurium test strains, TA98 and TA100 studies revealed that the degradation of crystal violet and malachite green by CM-4 did not lead to mutagenic products. Altogether, these results demonstrated the usefulness of the bacterial consortium in the treatment of the textile dyes. Jihane Cheriaa, Monia Khaireddine, Mahmoud Rouabhia, and Amina Bakhrouf Copyright © 2012 Jihane Cheriaa et al. All rights reserved. Removal of Cd(II) from Aquatic System Using Oscillatoria sp. Biosorbent Mon, 30 Apr 2012 11:23:02 +0000 Biosorption of Cd(II) ions from aqueous solutions by native and dried Oscillatoria sp. Cyanobacterium biomass was investigated in the batch mode. The Oscillatoria sp. was prepared from Molecular and Cell Laboratory of University of Mazandaran and grown in BG-11 medium. A comparison of Cd(II) adsorption properties of dried with native Oscillatoria sp. biomass was made, the dried one showed a higher biosorption capacity and faster kinetic. The influence of solution pH, contact time, biomass concentration, initial metal ion concentration, and presence of coions using dried Oscillatoria sp. biomass as well as pretreatment on the biosorption capacity of the biomass were studied. Various pretreatments of Oscillatoria sp. increased biosorption of Cd(II) at pH 7 in comparison with native biomass. However, heating at 100°C in a water bath showed significant improvement in Cd(II) biosorption capacity. The experimental biosorption data was well fitted to the Freundlich model compared to the Langmuir model, and the amount of Cd(II) removed from solution increased with increasing Cd(II) concentration. In addition, the dried biomass was investigated for Cd(II) removal from the simulated real sample containing about 14 mg/l Cd(II) at pH 7, under the same experimental condition. Seyed Naser Azizi, Abasalt Hosseinzadeh Colagar, and Seyede Maryam Hafeziyan Copyright © 2012 Seyed Naser Azizi et al. All rights reserved. Back to Natural Fiber: Wool Color Influences Its Sensitivity to Enzymatic Treatment Thu, 26 Apr 2012 15:44:55 +0000 There are many missed biotechnological opportunities in the developmental countries. Wool quality improvement is one of them. This study is concerning with improving the wool quality using technical enzymes. White wool proves to be more susceptible to the enzymatic treatment than blackish brown wool. This proves that the enzymatic reaction is sensitive to the natural color differences between wool fibers. A simple enzymatic method has been used to improve the wool quality as well as to investigate the changes happened in the wool fibers. Geobacillus stearothermophilus has been used under mesophilic and static cultivation conditions using wool as the main carbon source. These conditions prove to be more suitable for maintaining the fiber structure, less expensive, and reliable as an in-house biotechnological process that can be adapted everywhere. The enzyme activity in case of white wool was 4 Units/ml and for blackish brown wool was 1.5 Units/ml. Electron microscope has been used to evaluate the end result. By following the process included in this paper using probable microbial strain(s), the wool quality improvement can be applied globally and can add another value to the economy of the developmental countries. Amro A. Amara Copyright © 2012 Amro A. Amara. All rights reserved. Computational Methods to Work as First-Pass Filter in Deleterious SNP Analysis of Alkaptonuria Thu, 19 Apr 2012 13:49:41 +0000 A major challenge in the analysis of human genetic variation is to distinguish functional from nonfunctional SNPs. Discovering these functional SNPs is one of the main goals of modern genetics and genomics studies. There is a need to effectively and efficiently identify functionally important nsSNPs which may be deleterious or disease causing and to identify their molecular effects. The prediction of phenotype of nsSNPs by computational analysis may provide a good way to explore the function of nsSNPs and its relationship with susceptibility to disease. In this context, we surveyed and compared variation databases along with in silico prediction programs to assess the effects of deleterious functional variants on protein functions. In other respects, we attempted these methods to work as first-pass filter to identify the deleterious substitutions worth pursuing for further experimental research. In this analysis, we used the existing computational methods to explore the mutation-structure-function relationship in HGD gene causing alkaptonuria. R. Magesh and C. George Priya Doss Copyright © 2012 R. Magesh and C. George Priya Doss. All rights reserved. Characterization of eDNA from the Clinical Strain Acinetobacter baumannii AIIMS 7 and Its Role in Biofilm Formation Thu, 19 Apr 2012 13:31:00 +0000 Release of extracellular DNA (eDNA) was observed during in vitro growth of a clinical strain of Acinetobacter baumannii. Membrane vesicles (MV) of varying diameter (20–200 nm) containing DNA were found to be released by transmission electron microscopy (TEM) and atomic force microscopy (AFM). An assessment of the characteristics of the eDNA with respect to size, digestion pattern by DNase I/restriction enzymes, and PCR-sequencing, indicates a high similarity with genomic DNA. Role of eDNA in static biofilm formed on polystyrene surface was evaluated by biofilm augmentation assay using eDNA available in different preparations, for example, whole cell lysate, cell-free supernatant, MV suspension, and purified eDNA. Biofilm augmentation was seen up to 224.64%, whereas biofilm inhibition was 59.41% after DNase I treatment: confirming that eDNA facilitates biofilm formation in A. baumannii. This is the first paper elucidating the characteristics and role of eDNA in A. baumannii biofilm, which may provide new insights into its pathogenesis. Praveen K. Sahu, Pavithra S. Iyer, Amrita M. Oak, Karishma R. Pardesi, and Balu A. Chopade Copyright © 2012 Praveen K. Sahu et al. All rights reserved. The Future of Butyric Acid in Industry Thu, 19 Apr 2012 11:12:21 +0000 In this paper, the different applications of butyric acid and its current and future production status are highlighted, with a particular emphasis on the biofuels industry. As such, this paper discusses different issues regarding butyric acid fermentations and provides suggestions for future improvements and their approaches. Mohammed Dwidar, Jae-Yeon Park, Robert J. Mitchell, and Byoung-In Sang Copyright © 2012 Mohammed Dwidar et al. All rights reserved. Optimization of Acid Protease Production by Aspergillus niger I1 on Shrimp Peptone Using Statistical Experimental Design Thu, 19 Apr 2012 11:08:28 +0000 Medium composition and culture conditions for the acid protease production by Aspergillus niger I1 were optimized by response surface methodology (RSM). A significant influence of temperature, KH2PO4, and initial pH on the protease production was evaluated by Plackett-Burman design (PBD). These factors were further optimized using Box-Behnken design and RSM. Under the proposed optimized conditions, the experimental protease production (183.13 U mL−1) closely matched the yield predicted by the statistical model (172.57 U mL−1) with 𝑅2=0.914. Compared with the initial M1 medium on which protease production was 43.13 U mL−1, a successful and significant improvement by 4.25 folds was achieved in the optimized medium containing (g/L): hulled grain of wheat (HGW) 5.0; KH2PO4 1.0; NaCl 0.3; MgSO4(7H2O) 0.5; CaCl2 (7H2O) 0.4; ZnSO4 0.1; Na2HPO4 1.6; shrimp peptone (SP) 1.0. The pH was adjusted at 5 and the temperature at 30∘C. More interestingly, the optimization was accomplished using two cheap and local fermentation substrates, HGW and SP, which may result in a significant reduction in the cost of medium constituents. Rayda Siala, Fakher Frikha, Samiha Mhamdi, Moncef Nasri, and Alya Sellami Kamoun Copyright © 2012 Rayda Siala et al. All rights reserved. Identification and Characterization of Differentially Expressed Transcripts in the Gills of Freshwater Prawn (Macrobrachium rosenbergii) under Salt Stress Thu, 19 Apr 2012 11:03:28 +0000 The giant freshwater prawn, Macrobrachium rosenbergii, is an economically important species. It is a euryhaline shrimp, surviving in wide-range salinity conditions. A change in gene expression has been suggested as an important component for stress management. To better understand the osmoregulatory mechanisms mediated by the gill, a subtractive and suppressive hybridization (SSH) tool was used to identify expressed transcripts linked to adaptations in saline water. A total of 117 transcripts represented potentially expressed under salinity conditions. BLAST analysis identified 22% as known genes, 9% as uncharacterized showing homologous to unannotated ESTs, and 69% as unknown sequences. All the identified known genes representing broad spectrum of biological pathways were particularly linked to stress tolerance including salinity tolerance. Expression analysis of 10 known genes and 7 unknown/uncharacterized genes suggested their upregulation in the gills of prawn exposed to saline water as compared to control indicating that these are likely to be associated with salinity acclimation. Rapid amplification of cDNA ends (RACE) was used for obtaining full-length cDNA of MRSW-40 clone that was highly upregulated during salt exposure. The sequenced ESTs presented here will have potential implications for future understanding about salinity acclimation and/or tolerance of the prawn. Hirak Kumar Barman, Swagat Kumar Patra, Varsha Das, Shibani Dutta Mohapatra, Pallipuram Jayasankar, Chinmayee Mohapatra, Ramya Mohanta, Rudra Prasanna Panda, and Surya Narayan Rath Copyright © 2012 Hirak Kumar Barman et al. All rights reserved. Bioremediation of Agro-Based Pulp Mill Effluent by Microbial Consortium Comprising Autochthonous Bacteria Wed, 01 Feb 2012 12:26:55 +0000 Small-scale agro-based pulp and paper mills are characterized as highly polluting industries. These mills use Kraft pulping process for paper manufacturing due to which toxic lignified chemicals are released into the environment. Lack of infrastructure, technical manpower, and research and development facilities restricts these mills to recover these chemicals. Therefore, the chemical oxygen demand (COD) of the emanating stream is quite high. For solving the above problem, four bacteria were isolated from the premises of agro-based pulp and paper mill which were identified as species of Pseudomonas, Bacillus, Pannonibacter, and Ochrobacterum. These bacteria were found capable of reducing COD up to 85%–86.5% in case of back water and 65-66% in case of back water : black liquor (60 : 40), respectively, after acclimatization under optimized conditions (pH 6.8, temperature 35°C, and shaking 200 rpm) when the wastewater was supplemented with nitrogen and phosphorus as trace elements. Virendra Kumar, Purnima Dhall, Rita Kumar, Yogendra Prakash Singh, and Anil Kumar Copyright © 2012 Virendra Kumar et al. All rights reserved. Biodegradation of Sewage Wastewater Using Autochthonous Bacteria Wed, 04 Jan 2012 11:05:58 +0000 The performance of isolated designed consortia comprising Bacillus pumilus, Brevibacterium sp, and Pseudomonas aeruginosa for the treatment of sewage wastewater in terms of reduction in COD (chemical oxygen demand), BOD (biochemical oxygen demand) MLSS (mixed liquor suspended solids), and TSS (total suspended solids) was studied. Different parameters were optimized (inoculum size, agitation, and temperature) to achieve effective results in less period of time. The results obtained indicated that consortium in the ratio of 1 : 2 (effluent : biomass) at 200 rpm, 35°C is capable of effectively reducing the pollutional load of the sewage wastewaters, in terms of COD, BOD, TSS, and MLSS within the desired discharge limits, that is, 32 mg/L, 8 mg/L, 162 mg/L, and 190 mg/L. The use of such specific consortia can overcome the inefficiencies of the conventional biological treatment facilities currently operational in sewage treatment plants. Purnima Dhall, Rita Kumar, and Anil Kumar Copyright © 2012 Purnima Dhall et al. All rights reserved. A Critical View of Current State of Phytotechnologies to Remediate Soils: Still a Promising Tool? Wed, 04 Jan 2012 10:54:57 +0000 Phytotechnologies are often shown as an emerging tool to remediate contaminated soils. Research in this field has resulted in many important findings relating to plant and soil sciences. However, there have been scant private and public investments and little commercial success with this technology. Here, we investigate the barriers to the adoption of phytotechnologies and determine whether it is still a fertile area for future research. The terminology used in phytotechnologies includes a confusing mish-mash of terms relating to concepts and processes increasing the difficulty of developing a unique commercial image. We argue that the commercial success of phytotechnologies depends on the generation of valuable biomass on contaminated land, rather than a pure remediation technique that may not compare favourably with the costs of inaction or alternative technologies. Valuable biomass includes timber, bioenergy, feedstock for pyrolosis, biofortified products, or ecologically important species. Héctor M. Conesa, Michael W. H. Evangelou, Brett H. Robinson, and Rainer Schulin Copyright © 2012 Héctor M. Conesa et al. All rights reserved. Application of In Vitro Protein Expression to Human Prote Mon, 01 Jan 1900 00:00:00 +0000 Peter T. Beernink, Simone S. Krupka, Victoria Lao, George Martin, and Matthew A. Coleman Copyright © 2002 Peter T. Beernink et al. All rights reserved. Interaction Effect between Phosphorus and Zinc on their Availability in Soil in Relation to their Contents in Stevia (Stevia rebaudiana) Mon, 01 Jan 1900 00:00:00 +0000 A greenhouse experiment was conducted at the Indian Institute of Horticultural Research (IIHR), Bangalore to study the interaction effect between phosphorus and zinc on their availability in soil in relation to their contents in stevia (Stevia rebaudiana). The results show that the amount of available P and Zn content in soil has been found to increase initially and, thereafter, the amount of the same decreased with the progress of plant growth up to 60 days irrespective of treatments. The amount of P and Zn in soils showed an increase with their separate applications either as soil or foliar spray while that of the same value significantly decreased both in soils and plants due to their combined applications, suggesting a mutual antagonistic effect between Zn and P affecting each other’'s availability in soil and content in the stevia plant. Kuntal Das, Raman Dang, T. N. Shivananda, and Pintu Sur Copyright © 2005 Kuntal Das. All rights reserved. Multiple Roles of the Y Chromosome in the Biology of Drosophila melanogaster Mon, 01 Jan 1900 00:00:00 +0000 The X and Y chromosomes of Drosophila melanogaster were the first examples of chromosomes associated with genetic information. Thanks to the serendipitous discovery of a male with white eyes in 1910, T.H. Morgan was able to associate the X chromosome of the fruit fly with a phenotypic character (the eye color) for the first time. A few years later, his student, C.B. Bridges, demonstrated that X0 males, although phenotypically normal, are completely sterile. This means that the X chromosome, like the autosomes, harbors genes that control several phenotypic traits, while the Y chromosome is important for male fertility only. Notwithstanding its long history – almost 100 years in terms of genetic studies – most of the features of the Y chromosome are still a mystery. This is due to the intrinsic nature of this genetic element, namely, (1) its molecular composition (mainly transposable elements and satellite DNA), (2) its genetic inertia (lack of recombination due to its heterochromatic nature), (3) the absence of homology with the X (with the only exception of the nucleolar organizer), (4) the lack of visible phenotypes when it is missing (indeed, except for their sterility, X0 flies are normal males), and (5) its low density as for protein-coding sequences (to date, only 13 genes out of approximately 14,000 have been mapped on this chromosome in D. melanogaster, i.e., ~0.1% of the total). Nonetheless, a more accurate analysis reveals that this chromosome can influence several complex phenotypes: (1) it has a role in the fertility of both sexes and viability of males when over-represented; (2) it can unbalance the intracellular nucleotide pool; (3) it can interfere with the gene expression either by recruiting proteins involved in chromatin remodeling (PEV) or, to a higher extent, by influencing the expression of up to 1,000 different genes, probably by changing the availability of transcription factors; (4) it plays a major role (up to 50%) in the resistance to heat-induced male sterility; (5) it affects the behavior; and (6) it plays a role in genetic imprinting. In the present paper, all these Y-related phenotypes are described and a potential similarity with the human Y chromosome is drawn. Roberto Piergentili Copyright © 2010 Roberto Piergentili. All rights reserved. Asymmetric Cell Kinetics Genes: The Key to Expansion of Adult Stem Cells in Culture Mon, 01 Jan 1900 00:00:00 +0000 A singular challenge in stem cell research today is the expansion and propagation of functional adult stem cells. Unlike embryonic stem cells, which are immortal in culture, adult stem cells are notorious for the difficulty encountered when attempts are made to expand them in culture. One overlooked reason for this difficulty may be the inherent asymmetric cell kinetics of stem cells in postnatal somatic tissues. Senescence is the expected fate of a culture whose growth depends on adult stem cells that divide with asymmetric cell kinetics. Therefore, the bioengineering of strategies to expand adult stem cells in culture requires knowledge of cellular mechanisms that control asymmetric cell kinetics. The properties of several genes recently implicated to function in a cellular pathway(s) that regulates asymmetric cell kinetics are discussed. Understanding the function of these genes in asymmetric cell kinetics mechanisms may be the key that unlocks the adult stem cell expansion problem. James L. Sherley Copyright © 2002 James L. Sherley. All rights reserved. Rheology of Biopolymer Solutions and Gels Mon, 01 Jan 1900 00:00:00 +0000 Rheological techniques and methods have been employed for many decades in the characterization of polymers. Originally developed and used on synthetic polymers, rheology has then found much interest in the field of natural (bio) polymers. This review concentrates on introducing the fundamentals of rheology and on discussing the rheological aspects and properties of the two major classes of biopolymers: polysaccharides and proteins. An overview of both their solution properties (dilute to semi-dilute) and gel properties is described. David R. Picout and Simon B. Ross-Murphy Copyright © 2003 David R. Picout and Simon B. Ross-Murphy. All rights reserved. The Current Status of Microscopical Hair Comparisons Mon, 01 Jan 1900 00:00:00 +0000 Although the microscopical comparison of human hairs has been accepted in courts of law for over a century, recent advances in DNA technology have called this type of forensic examination into question. In a number of cases, post-conviction DNA testing has exonerated defendants who were convicted in part on the results of microscopical hair comparisons. A federal judge has held a Daubert hearing on the microscopical comparison of human hairs and has concluded that this type of examination does not meet the criteria for admission of scientific evidence in federal courts. A review of the available scientific literature on microscopical hair comparisons (including studies conducted by the Royal Canadian Mounted Police and the Federal Bureau of Investigation) leads to three conclusions: (1) microscopical comparisons of human hairs can yield scientifically defensible conclusions that can contribute to criminal investigations and criminal prosecutions, (2) the reliability of microscopical hair comparisons is strongly affected by the training of the forensic hair examiner, (3) forensic hair examiners cannot offer estimates of the probability of a match of a questioned hair with a hair from a randomly selected person. In order for microscopical hair examinations to survive challenges under the U.S. Supreme Court’s Daubert decision, hair microscopists must be better trained and undergo frequent proficiency testing. More research on the error rates of microscopical hair comparisons should be undertaken, and guidelines for the permissible interpretations of such comparisons should be established. Until these issues have been addressed and satisfactorily resolved, microscopical hair comparisons should be regarded by law enforcement agencies and courts of law as merely presumptive in nature, and all microscopical hair comparisons should be confirmed by nuclear DNA profiling or mitochondrial DNA sequencing. Walter F. Rowe Copyright © 2001 Walter F. Rowe. All rights reserved. Interaction Between Phosphorus and Zinc on the Biomass Yield and Yield Attributes of the Medicinal Plant Stevia (Stevia rebaudiana) Mon, 01 Jan 1900 00:00:00 +0000 A greenhouse experiment was conducted at the Indian Institute of Horticultural Research (IIHR), Bangalore to study the interaction effect between phosphorus (P) and zinc (Zn) on the yield and yield attributes of the medicinal plant stevia. The results show that the yield and yield attributes have been found to be significantly affected by different treatments. The total yield in terms of biomass production has been increased significantly with the application of Zn and P in different combinations and methods, being highest (23.34 g fresh biomass) in the treatment where Zn was applied as both soil (10 kg ZnSO4/ha) and foliar spray (0.2% ZnSO4). The results also envisaged that the different yield attributes viz. height, total number of branches, and number of leaves per plant have been found to be varied with treatments, being highest in the treatment where Zn was applied as both soil and foliar spray without the application of P. The results further indicated that the yield and yield attributes of stevia have been found to be decreased in the treatment where Zn was applied as both soil and foliar spray along with P suggesting an antagonistic effect between Zn and P. Kuntal Das, Raman Dang, T. N. Shivananda, and Pintu Sur Copyright © 2005 Kuntal Das et al. All rights reserved. Rapid Purification of Nuclei from Animal and Plant Tissues and Cultured Cells Mon, 01 Jan 1900 00:00:00 +0000 Nuclei are isolated by buoyant density banding in a discontinuous iodixanol gradient, under isoosmotic conditions. The low viscosity of the gradient allows the purification to be carried out at 10,000g in only 20 min. The method avoids possible damage to nucleoprotein complexes caused by hyperosmotic sucrose gradients. Although developed for mammalian liver the method can be applied (with or without minor modifications) to any tissue or cell type. John Graham Copyright © 2002 John Graham. All rights reserved. Effect of Putting Grip on Eye and Head Movements During the Golf Putting Stroke Mon, 01 Jan 1900 00:00:00 +0000 The objective of this article is to determine the effect of three different putting grips (conventional, cross-hand, and one-handed) on variations in eye and head movements during the putting stroke. Seven volunteer novice players, ranging in age from 21 to 22 years, participated in the study. During each experimental session, the subject stood on a specially designed platform covered with artificial turf and putted golf balls towards a standard golf hole. The three different types of grips were tested at two distances: 3 and 9 ft. For each condition, 20 putts were attempted. For each putt, data were recorded over a 3-s interval at a sampling rate of 100 Hz. Eye movements were recorded using a helmet-mounted eye movement monitor. Head rotation about an imaginary axis through the top of the head and its center-of-rotation was measured by means of a potentiometer mounted on a fixed frame and coupled to the helmet. Putter-head motion was measured using a linear array of infrared phototransistors embedded in the platform. The standard deviation (STD, relative to the initial level) was calculated for eye and head movements over the duration of the putt (i.e., from the beginning of the backstroke, through the forward stroke, to impact). The averaged STD for the attempted putts was calculated for each subject. Then, the averaged STDs and other data for the seven subjects were statistically compared across the three grip conditions. The STD of eye movements were greater (p < 0.1) for conventional than cross-hand (9 ft) and one-handed (3 and 9 ft) grips. Also, the STD of head movements were greater (p < 0.1; 3 ft) for conventional than cross-hand and one-handed grips. Vestibulo-ocular responses associated with head rotations could be observed in many 9 ft and some 3 ft putts. The duration of the putt was significantly longer (p < 0.05; 3 and 9 ft) for the one-handed than conventional and cross-hand grips. Finally, performance, or percentage putts made, was significantly better (p <0.05; 9 ft) for cross-hand than conventional grip. The smaller variations, both in eye movements during longer putts and head movements during shorter putts, using cross-hand and one-handed grips may explain why some golfers, based on their playing experience, prefer these over the conventional grip. Also, the longer duration for the one-handed grip, which improves tempo, may explain why some senior players prefer the long-shaft (effectively one-handed grip) putter. George K. Hung Copyright © 2003 George K. Hung. All rights reserved. Immobilization of Zinc Phthalocyanines in Silicate Matrices and Investigation of Their Photobactericidal Effect on E.coli Mon, 01 Jan 1900 00:00:00 +0000 The aim of the present investigation was to immobilize zinc phthalocyanines in a silicate matrix and to test the photobactericidal properties of the matrices so prepared toward Esherichia coli in model aqueous media. For the purpose, tetra tertiary butyl zinc phthalocyanine (TBZnPc) and zinc phthalocyanine tetrasulfonic acid (ZnPcTS) were used. The abilities of these two photosensitizers to generate singlet oxygen in solution were compared by following the rate of photobleaching of 1,3-diphenylisobenzofuran (DPBF) at 430 nm in dimethylformamide (DMF).The results of this study show clearly that, under the conditions used here, the TBZnPc is the more effective generator of singlet oxygen; with it the DPBF was virtually completely photobleached in 4 min, while with the ZnPcTS under the same conditions, it took 12 min to reach this point. Glass conjugates with the two phthalocyanines were obtained by the sol-gel technique and were characterized by a well-defined color due to the phthalocyanine incorporated in the silicate matrix. Glasses with an intense, but inhomogeneous, green color were obtained when the tetrasulfonic derivative of the zinc phthalocyanine was used, while blue glasses of evenly distributed coloration were formed from the tetra tertiary butyl derivative.The ZnPcTS conjugate demonstrates more effective singlet oxygen evolution than is the case with the TBZnPc conjugate. These results are the opposite of those obtained for the free phthalocyanines in solution. The structural formulae of the compounds show that TBZnPc has a more pronounced hydrophobic character than the sulfonic derivative. In our view, the relative reactivities of the conjugates can be explained by the tetrasulfonic derivative being situated mainly in the surface parts of the glass matrix where the hydrophilic character is prevailing, while the tertiary butyl derivative is mainly present in the internal parts of the matrix as a result of which it is less accessible and therefore less activeThe results obtained on the effect of zinc phthalocyanine conjugates on E. coli show a trend similar to that observed with singlet oxygen evolution shown. Thus, for the ZnPcTS conjugate, the log kill is 1.32 and for the TBZnPc conjugate, it is 0.98, in each case after 120 min. The results obtained show that phthalocyanines can be immobilized successfully in a silicate matrix and used for photodisinfection of microbially polluted waters. The silicate matrix has some advantages in comparison with other organic matrices. It is insoluble in water, resistant towards microorganisms, easy to fabricate, and might be developed successfully for the photodisinfection of water, e.g., in swimming pools and in other open water reservoirs. Spas Artarsky, Stanislava Dimitrova, Raymond Bonnett, and Milka Krysteva Copyright © 2006 Spas Artarsky et al. All rights reserved. Optical Enzyme Sensor for Urea Determination via Immobilized pH Indicator and Urease onto Transparent Membranes Mon, 01 Jan 1900 00:00:00 +0000 Transparent triacetylcellulose membranes with immobilized pH indicator (neutral red) as well as with simultaneously immobilized urease and neutral red were used as optical sensors for determination of urea concentrations in model solutions. Decomposition of urea with the enzyme urease is accompanied by evolution of ammonia. This leads to the changes of the neutral red absorption, which is proportional to the substrate (urea) within certain concentration limits in model solution. As a result of the investigation, standard curves were plotted for determination of urea over the range of 1 to 500 mM using immobilized indicator and free urease. Simultaneous immobilization of indicator and urease permitted determination of urea in the interval 50 to 500 mM. The membrane used contained 0.169 U urease activity on an area of 1.7 cm2. The standard curves were plotted using the linear region of the kinetic curves for the corresponding substrate concentrations. A possible scheme of the interaction between the activated triacetylcellulose membrane and the indicator and enzyme is proposed. The membranes obtained are suitable for repeated ecological applications where urea is to be determined. Milka Krysteva and Mohamed Al Hallak Copyright © 2003 Milka Krysteva and Mohamed Al Hallak. All rights reserved. Purification of Gst Fusion Proteins, On-Column Cleavage, and Sample Clean Up Mon, 01 Jan 1900 00:00:00 +0000 Jill A. Sigrell and Anna Heijbel Copyright © 2002 Jill A. Sigrell and Anna Heijbel. All rights reserved. Bioreactors for Decreasing the Growth of Brain Tumors Mon, 01 Jan 1900 00:00:00 +0000 Malignant gliomas are the most common primary brain tumors. They are highly aggressive tumors characterized by a recurrence rate of virtually 100%. Despite significant advances in neuroimaging and neurosurgical techniques, the median survival time of patients with glioblastoma multiforme remains 12 to 18 months. Malignant gliomas are characterized by rapidly dividing cells, which invade into the normal brain, and a high degree of vascularity. Recent experimental evidence indicates that tumor-related angiogenesis contributes significantly to the malignant phenotype. Rona S. Carroll and Marcelle Machluf Copyright © 2001 Rona S. Carroll and Marcelle Machluf. All rights reserved. HRAD30A, a Gene Encoding a Class Y Low-Fidelity Dna Polymerase Has a Low Complexity Region in Intron I Associated with Tissue-Specific Alternative Splicing Mon, 01 Jan 1900 00:00:00 +0000 James E. Cleaver, Manish Thakur, Stas Volik, and Colin Collins Copyright © 2002 James E. Cleaver et al. All rights reserved. Separation of Membrane Vesicles and Cytosol from Cultured Cells and Bacteria in a Preformed Discontinuous Gradient Mon, 01 Jan 1900 00:00:00 +0000 There are many situations when it is necessary to separate rapidly and efficiently a cytosolic and a membrane vesicle fraction from either cultured cells or from bacteria. Flotation of the vesicles through a low-density barrier from a dense sample zone using the low viscosity medium iodixanol allows complete separation of these compartments. As the sample is exposed to the gmax the tendency of the proteins to sediment overcomes any diffusion in the opposite direction. John Graham Copyright © 2002 John Graham. All rights reserved. Receptor-Mediated Gene Delivery Mon, 01 Jan 1900 00:00:00 +0000 Receptor-mediated gene delivery capitalises on the presence of specific cell surface molecules for DNA uptake into cells and represents a particularly appealing approach for targeting vector DNA to specific cell types in vivo and in vitro. Various ligand/DNA and antibody/DNA transfer complexes were generated that, following binding to cells, are internalised and reach the endosomal compartment. Vector complexes contain endosomolytic components that ensure vector release from the endosome and translocation of vector DNA into the nucleus where transcription occurs. Thus, receptor-mediated gene delivery encompasses several critical steps that must be considered when designing and applying such vector systems. Tatjana C. Gust and Martin Zenke Copyright © 2002 Tatjana C. Gust and Martin Zenke. All rights reserved. Bioremediation of Heavy Metals and Organic Toxicants by Composting Mon, 01 Jan 1900 00:00:00 +0000 Hazardous organic and metallic residues or by-products can enter into plants, soils, and sediments from processes associated with domestic, municipal, agricultural, industrial, and military activities. Handling, ingestion, application to land or other distributions of the contaminated materials into the environment might render harm to humans, livestock, wildlife, crops, or native plants. Considerable remediation of the hazardous wastes or contaminated plants, soils, and sediments can be accomplished by composting. High microbial diversity and activity during composting, due to the abundance of substrates in feedstocks, promotes degradation of xenobiotic organic compounds, such as pesticides, polycyclic aromatic hydrocarbons (PAHs), and polychlorinated biphenyls (PCBs). For composting of contaminated soils, noncontaminated organic matter should be cocomposted with the soils. Metallic pollutants are not degraded during composting but may be converted into organic combinations that have less bioavailability than mineral combinations of the metals. Degradation of organic contaminants in soils is facilitated by addition of composted or raw organic matter, thereby increasing the substrate levels for cometabolism of the contaminants. Similar to the composting of soils in vessels or piles, the on-site addition of organic matter to soils (sheet composting) accelerates degradation of organic pollutants and binds metallic pollutants. Recalcitrant materials, such as organochlorines, may not undergo degradation in composts or in soils, and the effects of forming organic complexes with metallic pollutants may be nonpermanent or short lived. The general conclusion is, however, that composting degrades or binds pollutants to innocuous levels or into innocuous compounds in the finished product. Allen V. Barker and Gretchen M. Bryson Copyright © 2002 Allen V. Barker and Gretchen M. Bryson. All rights reserved.