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Fig. 3 | BMC Biotechnology

Fig. 3

From: Protoplast transformation as a potential platform for exploring gene function in Verticillium dahliae

Fig. 3

Assay for siRNA inhibition of Vta2 gene. Protoplasts (200 μl of 106/ml) isolated from Vd-wt were transformed with 10 μM siRNA-vta1, siRNA-vta2, siRNA-vta3 and siRNA-vta4 independently in separate tubes and also with 2.5 μM each of these siRNAs via PEG-mediated transformation. Protoplasts were regenerated for 18 h in TB3 broth at 25 °C, pelleted at 2500 rpm for 5 min, then resuspended in 200 μl TB3 broth and cultured in the center of CM agar plate. Colony diameter was measured for each group after 7 days at 25 °C (Control, siRNA-vta1, siRNA-vta2, siRNA-vta3, siRNA-vta4 and siRNA-vtamix). a Colony morphology of different groups on CM agar. b Colony diameters of control and siRNA groups. c Relative expression level of Vta2 gene in different siRNA treated groups. RNA was isolated from mycelia harvested after 72 h, first strand cDNA was synthesized and qRT-PCR was conducted for different siRNA groups

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