Ramesh BhatAshlesha C. Patil2016-11-122016-11-122011http://krishikosh.egranth.ac.in/handle/1/85243Transgenic tomato plants carrying Sclerotium rolfsii lectin gene (srl1) were generated by Agrobacterium-mediated transformation. Of the 90 independent regenerants, 40 could amplify 429 bp product with srl1-specific PCR. Segregation analysis in three selected events indicated single copy insertion in SRL1-T0(7) and multi-copy integration in SRL1-T0(10) and SRL1-T0(21). In SRL1-T0(7), the TDNA/ srl1 insertion was in a non-genic region between 85,375 and 85,376 bp of the clone C11_HBa0054I23 (GenBank Acc. No. AC212431.2) of chromosome 11. Crude extract of SRL1-T0(7), SRL1-T0(10) and SRL1-T0(21) showed an extra band of 17 kDa on SDS-PAGE. This protein along with their partially purified forms could agglutinate rabbit erythrocytes, indicating lectin activity. Specific activity of the lectin was more in the partially purified samples compared to protein from the crude extracts. Bioassay with second stage juveniles (J2s) of Meloidogyne incognita, the root knot nematode showed that on 3rd day of inoculation the infection rate was 66.66, 83.33, 91.66 percent in SRL1-T0(7), SRL1-T0(10), SRL1-T0(21), respectively compared to 100 per cent infection in the non-transgenic plant. On 6th day, the percentage of vermiform J2s developed into sausage-form was higher in nontransgenic plants compared to transgenic lines. After 70 days of inoculation, nontransgenic plants showed an average gall number of 59.6 per plant whereas, it was reduced to 33.6, 43.2 and 40.8 galls/plant, respectively in SRL1-T0(7), SRL1-T0(10), SRL1-T0(21). In general, non-transgenic plants showed a gall index of 7, hence they were rated as “highly susceptible”, whereas transgenic lines with a gall index of 3 were classified as “moderately resistant”. Nematode infection also resulted in significantly reduced average shoot and root length, and biomass in non-transgenic plants compared to transgenic plants. Transgenic plants were evaluated for white fly (Bemisia tabaci) mortality. It was 24%, 22% and 16% in SRL1-T0(7), SRL1-T0(10), SRL1-T0(21), respectively, which were higher than that found in non-transgenic plant (10%).Plant BiotechnologyThesis