Journal of Environmental Biology

Establishment of Glyoxalase I gene transformation and regeneration of cotton (Gossypium hirsutum L.)

A. Muthusamy1, 2* 

¹Plant Genetic Manipulation Lab, School of Life Sciences, Jawaharlal Nehru University, New Delhi – 110 067, India

²Present address: Department of Plant Sciences, Manipal School of Life Sciences, Manipal Academy of Higher Education, Manipal – 576 104, India

*Corresponding Author Email : amsamy.slsmu@gmail.com

Received: 28.07.2019                                                                    Revised: 16.12.2019                                            Accepted: 23.10.2020

Abstract

Aim: The current study was carried out to develop transgenic cotton plantlets with glyoxalase I (gly I) gene using Agrobacterium-mediated transformation.

Methodology: Seeds of cotton were inoculated on MS medium and the explants such as shoot tip (3-5 mm), hypocotyl and leaf were aseptically removed from in vitro plantlets. The pre-cultured and infected explants with Agrobacterium harboring gly I gene and the shoot tip were inoculated on MS media for shoot initiation and subcultured on elongation medium with growth hormones, and antibiotics. Healthy and well-grown shoots were subcultured on medium with indole butyric acid (IBA) (0.3 mgl-1) for root formation and the plantlets were hardened in plastic cups with sterile soil. The putative transgenic plantlets were analyzed histochemically for gus gene and the PCR analysis was performed for gly 1 gene.

Results: The transformation efficiency of cotton ranged 48.57 to 64.53 %. The regenerated plantlets showed the presence of gus gene in terms of blue coloration in shoots, whole leaf and leaf segments. The PCR was performed in putative transgenic plant lets with both gus gene as well as gly I gene primers. The PCR results showed the presence of 1031 bp DNA band with gus gene primers and 800 bp DNA band with the gly I gene primers.               

Interpretation: The current study has proven the reproducible procedure for the Agrobacterium-mediated gene transfer and regeneration of Indian cotton varieties. The PCR results revealed the presence of glyoxalase I gene in the transformants.      

Key words: Cotton varieties, Glyoxalase I gene, PCR analysis, Regeneration, Transformation

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