Life Sciences & Biotechnology
Title : | Evaluation of insecticidal activity for novel recombinant Vip3 proteins from newly isolated Bacillus thuriengenesis against lepidopteran pests and their efficacy in plant system for transgeneic approach. |
Area of research : | Life Sciences & Biotechnology |
Principal Investigator : | Dr. Lakshmikanth Mariyanna, University of Agricultural Sciences, Raichur, Karnataka |
Timeline Start Year : | 2023 |
Timeline End Year : | 2026 |
Contact info : | mlakshmikanth@uasraichur.edu.in |
Equipments : | Upright Deep Freezer (-80 degree celcius)
TAble top High speed cold centrifuge
Refrigerator double door (-4 degree)
Upright Depp freezer (-20 degree celcius) |
Details
Executive Summary : | In addition to the Cry proteins, certain strains of Bacillus thuringiensis (Bt) produce vegetative insecticidal proteins (Vips) and that have gained a huge interest as novel insecticides due to evolution of Cry protein resistant pests due to long term exposure. Among three classes of Vip proteins reported, Vip3 is toxic to a wide variety of members of the lepidoptera species. Further, as the mechanism of action of Vip3 and Cry proteins differ, there could be very low levels of cross-resistance suggesting Vip3 proteins as an alternative non-synthetic insecticide to overcome the problem of insect resistance. So far, 132 Vip3 proteins have been reported worldwide with 45 % to 78 % of similarity in their protein sequence and varying percentatge of toxicity against pests. Thus, Vip3 proteins are considered as a promising new generation of pest killers as bioinsecticides that can be used in spray formulation or by generation of transgenic crops to target broad spectrum of insectpests. In this regard, we have initiated the project by isolating several Bt strains from native soil samples collected from the different cropping system and different soil types and . 35 isolates have been identified as Bacillus identified by 16s RNA sequencing. Those 35 isolates were screened for the presence of Vip3 genes by colony PCR and 14 isolates of out them showed the presence of Vip3 gene. Initial bioassays with with 14 isolates against Plutella Xylostella, six isoaltes showed 100 % mortality within 48 hours of treatment. The Vip3 genes from these isolates were cloned and sequence confirmed. Upon sequence analysis and bioinformatic studies they are identified as novel Vip3 genes. Thus in the present proposal we aim to generate recombinant Vip3 proteins by cloning and overpressing it in the E.coli and Arabidopsis plant expression system. The insecticidal activity of the recombinant proteins will be tested against major lepidopteran pests (Pink bollworm, fall army worm, cotton bollworm, and Dimond back moth). |
Total Budget (INR): | 48,34,776 |
Organizations involved