Eurotium Cristatum Postfermentation of Fireweed and Apple Tree Leaf Herbal Teas

Table 1

Studied DNA regions of the fungus and primers and PCR annealing temperature.

DNA regions/genes	Symbols	Thermocycling programs	Primers (5→3)
			Symbols	Sequences
Entire ITS rDNA and fragment of 28S rDNA	ITS-D1/D2	(1) 94°C–5 min; (2) 33 cycles with temperature interval; 94°C–1 min, 51°C–1 min, 72°C–1 min; (3) 72°C–7 min	ITS1f	TCCGTAGGTGAACTTGCG
			NL4	GGTCCGTGTTTCAAGG
Large subunit 28S of ribosomal DNA	LSU	(1) 94°C–5 min; (2) 33 cycles with temperature interval; 94°C–1 min, 49°C–1 min, 72°C–2 min; (3) 72°C–7 min	LR0R	ACCCGCTGAACTTAAGC
			LR5	TCCTGAGGGAAACTTCG
RNA polymerase II gene	RPB2	(1) 94°C–5 min; (2) 9 cycles 94°C–1 min, at 60°C to 50°C–1 min (with 1 degree decrement each cycle), 72°C–1.5 min; (3) 32 cycles 94°C–1 min, 50°C–1.5 min, 72°C–1.5 min; (4) 72°C–7 min	fRPB2-5F	GAYGAYMGWGATCAYTTYGG
			fRPB2-7cR	CCAT(AG)GCTTG(CT)TT(AG)CCCAT
Translation elongation factor 1-α gene	TEF1-α	(1) 94°C–5 min; (2) 9 cycles 94°C–1 min, at 66°C to 56°C–1 min (with 1 degree decrement each cycle), 72°C–1.5 min; (3) 32 cycles 94°C–1 min, 56°C–1.5 min, 72°C–1.5 min; (4) 72°C–7 min	EF-595F	CGTGACTTCATCAAGAACATG
			EF-1567R	ACHGTRCCRATACCACCRATCTT
β-tubulin gene	β-tub	(1) 94°C–5 min; (2) 9 cycles 94°C–1 min, at 64°C to 54°C–1 min (with 1 degree decrement each cycle), 72°C–1.5 min; (3) 32 cycles 94°C–1 min, 54°C–1.5 min, 72°C–1.5 min; (4) 72°C–7 min	βt2a	GGTAACCAAATCGGTGCTGCTTTC
			βt2b	ACCCTCAGTGTAGTGACCCTTGGC
Glycerol 3-phosphate dehydrogenase gene	GAPDH	(1) 94°C–5 min; (2) 9 cycles 94°C–1 min, at 65°C to 55°C–1 min (with 1 degree decrement each cycle), 72°C–1.5 min; (3) 32 cycles 94°C–1 min, 55°C–1.5 min, 72°C–1.5 min; (4) 72°C–7 min	gpd1	CAACGGCTTCGGTCGCA TTG
			gpd2	GCCAAGCAGTTGGTTGTGC