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Evaluation of transgenic chickpea harboring codon-modified Vip3Aa against gram pod borer (Helicoverpa armigera H.)

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 Lepi...

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Autores principales: Singh, Prateek, G. K., Sujayanand, Thakur, Shallu, Rathore, Meenal, Verma, Om Prakash, Singh, Narendra Pratap, Das, Alok
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9231776/
https://www.ncbi.nlm.nih.gov/pubmed/35749522
http://dx.doi.org/10.1371/journal.pone.0270011
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author Singh, Prateek
G. K., Sujayanand
Thakur, Shallu
Rathore, Meenal
Verma, Om Prakash
Singh, Narendra Pratap
Das, Alok
author_facet Singh, Prateek
G. K., Sujayanand
Thakur, Shallu
Rathore, Meenal
Verma, Om Prakash
Singh, Narendra Pratap
Das, Alok
author_sort Singh, Prateek
collection PubMed
description 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.
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spelling pubmed-92317762022-06-25 Evaluation of transgenic chickpea harboring codon-modified Vip3Aa against gram pod borer (Helicoverpa armigera H.) Singh, Prateek G. K., Sujayanand Thakur, Shallu Rathore, Meenal Verma, Om Prakash Singh, Narendra Pratap Das, Alok PLoS One Research Article 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. Public Library of Science 2022-06-24 /pmc/articles/PMC9231776/ /pubmed/35749522 http://dx.doi.org/10.1371/journal.pone.0270011 Text en © 2022 Singh et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Singh, Prateek
G. K., Sujayanand
Thakur, Shallu
Rathore, Meenal
Verma, Om Prakash
Singh, Narendra Pratap
Das, Alok
Evaluation of transgenic chickpea harboring codon-modified Vip3Aa against gram pod borer (Helicoverpa armigera H.)
title Evaluation of transgenic chickpea harboring codon-modified Vip3Aa against gram pod borer (Helicoverpa armigera H.)
title_full Evaluation of transgenic chickpea harboring codon-modified Vip3Aa against gram pod borer (Helicoverpa armigera H.)
title_fullStr Evaluation of transgenic chickpea harboring codon-modified Vip3Aa against gram pod borer (Helicoverpa armigera H.)
title_full_unstemmed Evaluation of transgenic chickpea harboring codon-modified Vip3Aa against gram pod borer (Helicoverpa armigera H.)
title_short Evaluation of transgenic chickpea harboring codon-modified Vip3Aa against gram pod borer (Helicoverpa armigera H.)
title_sort evaluation of transgenic chickpea harboring codon-modified vip3aa against gram pod borer (helicoverpa armigera h.)
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9231776/
https://www.ncbi.nlm.nih.gov/pubmed/35749522
http://dx.doi.org/10.1371/journal.pone.0270011
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