Transformation of Mungbean Plants for Salt and Drought Tolerance by introducing a gene for an osmoprotectant glycine betaine

Anila Baloda,P.K.J Aiwal,Seema Madanpotra

doi:10.19071/jpsp.2017.v3.3148

Abstract

Mungbean (VignaradiataL. Wilczek) is an important grain legume widely cultivated in tropical and subtropical regions of the Indian subcontinent and in South East Asian countries. Protein and carbohydrate of Mungbean are easily digestible and create less flatulence than proteins derived from other legumes.Mungbean is very sensitive to salinity, drought, high and low temperature during the flowering and seed/pod development stages resulting in heavy losses to productivity. The development of genetically engineered plants by the introduction and/or over expression of selected abiotic stress tolerant genes seems to be a viable option for obtaining improved plants. Stable transformation and expression of transgene (codA gene) was achieved in mungbean through Agrobacterium tumefaciensmediated system using cotyledonary node explants, under the optimized conditions. Molecular analysis of transgenic plants was done by using PCR, DOT-BLOT, ELISA and Western blotting. The primary transformants were checked for salt tolerance by the leaf disc test.

Highlights

The different approaches used by many plants and microorganisms to defend themselves with abiotic stress are to synthesis and accumulation of compounds known as osmoprotectants, generally, we called them compatible solutes
GB accumulates under stress conditions in many plants which are known as natural accumulator of GB and these plants belongs to the families, Chenopodiaceae and Poaceae
40 healthy plantlets were shifted to soil, and out of them, 25 plants survived.These plants were morphologically similar to seed-raised plants

Summary

Introduction

Genetic transformation is a beneficial way for introducing genes from far-distant genomes, or from prokaryotes to eukaryotes and from the sources other than plants. the recent progress of transformation approach for producing abiotic stress resistance in plants has been reviewed by Sharma and Lavanya (2002), Wang et al (2003), Flowers (2004), Vinocur and Altman, (2005), Chinnusamy et al (2006), Yamaguchi and Blumwald (2005), Blumwald and Grover (2006), BhatnagarMathur (2008).Various transgenic strategies used so far includes the transformation of the plants through genes that conceal for an important enzyme(s)/protein(s) involved in ion/proton transport, biosynthesis of certain osmoprotectants (Garg, et al, 2002), scavengers of reactive oxygen species, proteins released during stress conditions like late-embryogenesis abundant proteins, signaling proteins in model plants such as arabidopsis, (Sakamoto, et al, 2000) Rice, (Sakamoto, et al, 1998, 2000; Mohanty et al 2002; Kathuria et al 2009),Tobacco (Rathinasabapathi et al, 1994, 2000; Holmstrom, et al Journal of Plant Stress Physiology ● Vol 3 ● 20172000; Huang, et al 2000 ) and Tomato (Park et al 2007; Goel et al 2011) rice, and tobacco.The different approaches used by many plants and microorganisms to defend themselves with abiotic stress are to synthesis and accumulation of compounds known as osmoprotectants, generally, we called them compatible solutes. For dot-blot analysis, the DNA isolated from transformed and nontransformed plants was denatured by heating on a temperature 95°C for 5-7 min and cooled rapidly on ice. Dots of DNA were made with the help of a micropipette on nylon membrane.

Results

Conclusion