Development of marker-free transgenic pigeon pea (Cajanus cajan) expressing a pod borer insecticidal protein

Abstract Pigeon pea, a grain legume of the semiarid tropics, is a rich source of high-quality protein. The productivity of this pulse is seriously affected by lepidopteron insect pests. To generate a sustainable insect-resistant plant, synthetically prepared bioactive key constituents of a crystal p...

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Autores principales:	Snehasish Sarkar, Souri Roy, Sudip K. Ghosh
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/088c6f3bba4f4f919c10e6e57dc13840
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spelling	oai:doaj.org-article:088c6f3bba4f4f919c10e6e57dc138402021-12-02T15:45:31ZDevelopment of marker-free transgenic pigeon pea (Cajanus cajan) expressing a pod borer insecticidal protein10.1038/s41598-021-90050-82045-2322https://doaj.org/article/088c6f3bba4f4f919c10e6e57dc138402021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-90050-8https://doaj.org/toc/2045-2322Abstract Pigeon pea, a grain legume of the semiarid tropics, is a rich source of high-quality protein. The productivity of this pulse is seriously affected by lepidopteron insect pests. To generate a sustainable insect-resistant plant, synthetically prepared bioactive key constituents of a crystal protein (Syn Cry1Ab) of Bacillus thuringiensis were expressed in pigeon pea under the guidance of a tissue-specific promoter of the RuBP carboxylase/oxygenase small subunit (rbcS) gene. Regenerated transgenic plants with the cry1Ab expression cassette (cry1Ab-lox-bar-lox) showed the optimum insect motility rate (90%) in an in vitro insect bioassay with second instar larvae, signifying the insecticidal potency of Syn Cry1Ab. In parallel, another plant line was also generated with a chimaeric vector harbouring a cre recombinase gene under the control of the CaMV 2 × 35S promoter. Crossing between T1 plants with a single insertion of cry1Ab-lox-bar-lox T-DNA and T1 plants with moderate expression of a cre gene with a linked hygromycin resistance (hptII) gene was performed to exclude the bialaphos resistance (bar) marker gene. Excision of the bar gene was achieved in T1F1 hybrids, with up to 35.71% recombination frequency. Insect-resistant pigeon pea plants devoid of selectable marker genes (syn Cry1Ab- bar and cre-hptII) were established in a consecutive generation (T1F2) through genetic segregation.Snehasish SarkarSouri RoySudip K. GhoshNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-16 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Snehasish Sarkar Souri Roy Sudip K. Ghosh Development of marker-free transgenic pigeon pea (Cajanus cajan) expressing a pod borer insecticidal protein
description	Abstract Pigeon pea, a grain legume of the semiarid tropics, is a rich source of high-quality protein. The productivity of this pulse is seriously affected by lepidopteron insect pests. To generate a sustainable insect-resistant plant, synthetically prepared bioactive key constituents of a crystal protein (Syn Cry1Ab) of Bacillus thuringiensis were expressed in pigeon pea under the guidance of a tissue-specific promoter of the RuBP carboxylase/oxygenase small subunit (rbcS) gene. Regenerated transgenic plants with the cry1Ab expression cassette (cry1Ab-lox-bar-lox) showed the optimum insect motility rate (90%) in an in vitro insect bioassay with second instar larvae, signifying the insecticidal potency of Syn Cry1Ab. In parallel, another plant line was also generated with a chimaeric vector harbouring a cre recombinase gene under the control of the CaMV 2 × 35S promoter. Crossing between T1 plants with a single insertion of cry1Ab-lox-bar-lox T-DNA and T1 plants with moderate expression of a cre gene with a linked hygromycin resistance (hptII) gene was performed to exclude the bialaphos resistance (bar) marker gene. Excision of the bar gene was achieved in T1F1 hybrids, with up to 35.71% recombination frequency. Insect-resistant pigeon pea plants devoid of selectable marker genes (syn Cry1Ab- bar and cre-hptII) were established in a consecutive generation (T1F2) through genetic segregation.
format	article
author	Snehasish Sarkar Souri Roy Sudip K. Ghosh
author_facet	Snehasish Sarkar Souri Roy Sudip K. Ghosh
author_sort	Snehasish Sarkar
title	Development of marker-free transgenic pigeon pea (Cajanus cajan) expressing a pod borer insecticidal protein
title_short	Development of marker-free transgenic pigeon pea (Cajanus cajan) expressing a pod borer insecticidal protein
title_full	Development of marker-free transgenic pigeon pea (Cajanus cajan) expressing a pod borer insecticidal protein
title_fullStr	Development of marker-free transgenic pigeon pea (Cajanus cajan) expressing a pod borer insecticidal protein
title_full_unstemmed	Development of marker-free transgenic pigeon pea (Cajanus cajan) expressing a pod borer insecticidal protein
title_sort	development of marker-free transgenic pigeon pea (cajanus cajan) expressing a pod borer insecticidal protein
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/088c6f3bba4f4f919c10e6e57dc13840
work_keys_str_mv	AT snehasishsarkar developmentofmarkerfreetransgenicpigeonpeacajanuscajanexpressingapodborerinsecticidalprotein AT souriroy developmentofmarkerfreetransgenicpigeonpeacajanuscajanexpressingapodborerinsecticidalprotein AT sudipkghosh developmentofmarkerfreetransgenicpigeonpeacajanuscajanexpressingapodborerinsecticidalprotein
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Development of marker-free transgenic pigeon pea (Cajanus cajan) expressing a pod borer insecticidal protein

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