Leaf-Encapsulated Vaccines: Agroinfiltration and Transient Expression of the Antigen Staphylococcal Endotoxin B in Radish Leaves

Figure 2

Agroinfiltration of SEB-GUS and TetC-GUS genes into radish leaves. (a) Syringe infiltration of A. tumefaciens into radish leaves. Bar = 6 mm. (b) Histochemical staining of radish leaves agroinfiltrated with A. tumefaciens (5 × 10⁷ CFU) containing no construct (as a control), 35S::SEB-GUS construct, or both 35S::SEB-GUS plus 35S::TetC-GUS constructs. To simultaneously express two antigens in planta, the left half of a single radish leaf was infiltrated with A. tumefaciens harboring a 35S::SEB-GUS construct while the right half was infiltrated with A. tumefaciens harboring a 35S::TetC-GUS construct. (c) For higher throughput antigen expression, six isolated radish leaves were concurrently infiltrated with nontransformed A. tumefaciens (number 1) as a control and A. tumefaciens carrying a 35S::SEB-GUS construct (numbers 2–6) using a multichannel pipette. Bar = 12 mm. (d) Histochemical staining of leaves simultaneously infiltrated using the multichannel pipette to indicate GUS expression one day after pipetting. Bar = 6 mm.

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