Engineering Resistance in Brinjal against Nematode {Meloidogyne Incognita) Using crylAb Gene from Bacillus Thuringiensis Berliner

摘要

Transgenic brinjal plants (Solanurn melongena cv. Co2) expressing crylAb gene from Bacillus thuringiensis Berliner (Bt) were evolved by Agrobacterium-mediated transformation system. In the present study, 7 To lines (P-Co2IAb-5, P-Co2IAb-9, P-Co2IAb-10, P-Co2IAb-12, P-Co2IAb-16, P-Co2IAb-17 and P-Co2IAb-22) harboring crylAb gene were obtained. PCR and Southern blotting analyses proved the presence and integration of crylAb in these lines. A polyclonal rabbit anti-CrylAb antiserum detected the expression of a 65 kDa CrylAb polypeptide in all these lines. Expression of CrylAb protein in those transgenic lines was also confirmed by lateral flow sticks (CrylAb) assay. Molecular analyses in T, progenies harboring crylAb gene proved the presence and expression of the transgene of interest. The chi-square (X~2) tests revealed a 3:1 segregation ratio for crylAb gene in the lines viz., P-Co2IAb-5 and P-Co2IAb-9, indicating that the transgene was integrated at a single locus. The transgenic lines thus obtained in this study were used to investigate the potential of the CrylAb protein in controlling the root knot nematode, Meloidogyne incognita. Bioassay for root knot nematode resistance in T, plants expressing CrylAb showed that there was a significant decrease in number of galls and eggmasses in crylAb-transgenic lines (P-Co2IAb-5 and P-Co2IAb-9) as compared to control. In addition, there was a significant decrease in the number of eggs per eggmass on those transgenic lines tested than the non-transgenic controls. Thus, transgenic B/-brinjal plants could also form a part of an integrated nematode management program.