Prateek Singh, Sujayanand G. K., Shallu Thakur, Meenal Rathore, Om Prakash Verma, Narendra Pratap Singh, Alok Das

Plos One; Published: June 24, 2022; https://doi.org/10.1371/journal.pone.0270011

Abstract

The gram pod borer is a major pest of chickpea, accounting for average annual yield losses to the tune of 40–50%. VIP3Aa, a class of insecticidal protein with different receptor binding site in the insect’s midgut compared to Bt-crystal protein, offers an alternative protection strategy against Lepidopteran insects. Here, we report evaluation of genetically engineered chickpea lines harboring codon modified Vip3Aa (cmVip3Aa) against the Lepidopteran insect pest, gram pod borer. The synthetic codon modified, cmVip3Aa gene of 2,370 bp was sub-cloned in modified plant expression vector and used for direct transformation of embryonic axis explants of chickpea (cv. DCP 92–3), with transformation efficiency of 4.30%. Presence and transmission of transgene across two generations were confirmed by PCR and Southern blot analyses in the five selected transgenic chickpea lines. Real Time PCR analyses indicated variable levels of cmVip3Aa expression in the transgenic chickpea lines (average Cq values 15.01±0.86 to 19.32±0.10), which were absent in the non-transgenic counterpart. Detached leaf insect bioassay indicate larval mortality (up to 39.75%), reduced larval feeding (up to 82.91%) and reduced larval weight gain (up to 68.23%), compared to control lines. Evaluation of gene offers a platform to identify efficacious insecticidal gene that can be used for insect resistance management in chickpea.

See https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0270011

Fig 1. Modified plant expression vector pRI 201-AN harboring cmVip3Aa.