Fig. 6

(i) in vitro bioassay with total soluble protein (20 µg) from DR-T-22 and untransformed Pusa basmati 1 plants; approximately, 10³ spores of M. oryzae (100 μl) suspension were germinated on a glass slide with 20 μg of total protein from leaf tissue of transgenic plants. Growth inhibition was assayed after 24 h, in terms of percent germination or altered morphology of hyphae as compared to the control samples. The treated and untreated samples were fixed and stained with 0.3 g/l calcofluor white examined under epifluorescence microscope. a M. oryzae treated with total soluble protein from DR-T-22 plant led to bulb formation at fungal hyphal tip. b M. oryzae treated with total soluble protein from untransformed plants showed uniform distribution of chitin and glucan and the septa were distinctly visible. (ii) Disease response of transformed and untransformed rice leaves challenged with M. oryzae spores. Twenty-one-day-old transgenic lines and untransformed control plants were inoculated with M. oryzae spores. Transverse sections of DR-T-22 transgenic lines and untransformed control were prepared. The samples were stained with calcofluor white. a Transverse sections of transgenic leaves with its corresponding bright field image. b Untransformed control with its corresponding bright field image. (iii) M. oryzae mycelia when treated with total protein from leaf tissue of transgenic plants showed abnormal morphology. Bar corresponds to 10 µm. a M. oryzae treated with total soluble protein from DR-T-22 plants showed hyper-branching; b M. oryzae treated with total soluble protein from untransformed plants showed profuse growth.