Fig. 12

ClToxB silencing. a RNA-silencing vector pSilent-Dual 1 (Nguyen et al. [55]). It contains two convergent promoters (Pgpd and PtrpC) of Aspergillus nidulans. ClToxB (JZ350031) was cloned in-between Pgpd and PtrpC promoters of pSilent-Dual 1, which was then used to transform Colletotrichum lentis isolate CT-21. b ClToxB expression in the resulting transformants (SToxB-1, SToxB-8 and SToxB-9) was determined by RT-qPCR. SToxB-8 displayed only 5 % of the ClToxB expression. c Susceptible lentil cultivar Eston was used to determine the anthracnose causing ability of silenced strains. Quantitative difference in virulence on Eston was observed among silenced strains at 6 days after inoculation (dai) and was clearly evident in the SToxB-infected plants. Disease scores were reported as least square means ± standard error. d Infected tissues collected at 3 dai were visualized under a light microscope. Quantitative difference in virulence was associated with the in planta fungal proliferation of silenced strains and the ClToxB expression level. A, Appressorium; PH, Primary biotrophic hyphae; and SH, Secondary necrotrophic hyphae. Bar =20 μM