Overexpression of ICE1 gene in mungbean ( Vigna radiata L.) for cold tolerance
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ORIGINAL ARTICLE
Overexpression of ICE1 gene in mungbean (Vigna radiata L.) for cold tolerance Gyana Ranjan Rout1 · Anuradha Bansal2 · Dhaneswar Swain1 · Kundansigh Rajpalsingh Jadhao1 · Rahul G. Shelke3 · Sanjib Kumar Panda4 Received: 8 August 2020 / Accepted: 1 October 2020 © Springer Nature B.V. 2020
Abstract The putative transgenic mungbean (Vigna radiata Var. OBGG-52) plants were regenerated on Murashige and Skoog (1962) medium supplemented with 1.0 mg/L 6-benzylaminopurine (BA). The transgenic status of transformants (T2 generation) was determined through PCR analysis using nptII and ICE1 gene-specific primers as well as uidA reporter gene expression. About 75% of the transformed plantlets were successfully established in the greenhouse. Overexpression of ICE1 gene in transgenic mungbean plants resulted in cold-tolerance at the seedling stage when compared to non-transformed plants. Seeds from T2 generation were further studied for cold tolerance through nutrient culture. Transgenic seeds had significantly increased germination and growth on a nutrient medium at 10–14 °C in growth chamber compared to the non-transgenic control. Moreover, the root and shoot length of transformed plants were significantly increased at 10–14 °C as compared with the non-transformed plants. Under cold stress, it was also observed that the transformed plantlets have significantly higher chlorophyll, proline, and lipid content as well as anti oxidative enzyme activity as compared to non-transformed plants. This study will help in mungbean improvement program for abiotic stress tolerance. Key message Cold temperature damaged the crop productivity of mungbean (Vigna radiata L. Wilczek), a high protein rich legume crop. The overexpression of cold responsive transcription factor, INDUCER OF CBF EXPRESSION 1 (ICE1) in mungbean enhanced cold tolerance. Keywords Vignar adiata · In vitro · Genetic transformation · ICE1 expression · Cold tolerance
Introduction
Communicated by Sergio J. Ochatt. * Gyana Ranjan Rout [email protected] 1
Department of Agricultural Biotechnology, College of Agriculture, Odisha University of Agriculture & Technology, Bhubaneswar, Odisha 751003, India
2
Biointeractions and Crop Protection, Rothamsted Research, West Common, Harpenden, Hertfordshire AL5 2JQ, UK
3
Department of Biosciences and Bioengineering, Indian Institute of Technology, Guwahati 781039, India
4
Department of Biochemistry, Central University of Rajasthan, Ajmer 305817, India
Mungbean, a short duration seasonal legume crop, is categorized under the Vigna genus and sub-genus Ceratotropis (Lambrides and Godwin 2007). It is cultivated in India, South East, and East Asia. The area under cultivation of this crop is more than 6 million hectares worldwide (about 8.5% of the global pulse area) and about 3 million tons of grain are produced annually. India is the major mungbean producing country where the cultivation area is around 3.5 million hectares and annual production is 1.2 million tons of grain (Nair et al. 2013). Mungbean
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