Enhancement of nematicidal potential through cloning and expression of gene from subsp. BTN7A strain.

A gene encoding chitinase from has been isolated after optimization of PCR conditions. It was cloned with two different prometers, T7 promoter of the pJET1.2/blunt cloning vector and the SP6 promoter of pGEM®-T Easy vector. After transforming DH5α, two transformants were selected, CHI-NRC-4 from the first vector and T-CHI-NRC-6 from the second vector, and used for further studies. The complete CDS sequence of gene was determined and submitted to GenBank with the accession number KX268692.1. Culture supernatants of (CHI-NRC-4) and (T-CHI-NRC-6) were investigated for their inhibitory effect on egg hatch under laboratory conditions. Result showed up to 96% inhibition in egg hatching due to both transformants as compared to control which reflect the same expression efficiency of both used prometers. A greenhouse experiment was carried out to evaluate the nematicidal effect of culture supernatants of the two transformts (CHI-NRC-4) and (T-CHI-NRC-6) against infected eggplant. Obtained results showed a significant reduction in nematode population in soil and roots and enhancement in eggplant growth parameters as compared to control.