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Abstract
Transgenic alfalfa (Medicago sativa L.) plants overexpressing the Arabidopsis ATP sulfurylase gene were generated using Agrobacterium-mediated genetic transformation to enhance their heavy metal accumulation efficiency. The ATP sulfurylase gene was cloned from Arabidopsis, following exposure to vanadium (V) and lead (Pb), and transferred into an Agrobacterium tumefaciens binary vector. This was co-cultivated with leaf explants of the alfalfa genotype Regen SY. Co-cultivated leaf explants were cultured on callus and somatic embryo induction medium, followed by regeneration medium for regenerating complete transgenic plants. The transgenic nature of the plants was confirmed using PCR and southern hybridization. The expression of Arabidopsis ATP sulfurylase gene in the transgenic plants was evaluated through RT-PCR. The selected transgenic lines showed increased tolerance to a mixture of five heavy metals and also demonstrated enhanced metal uptake ability under controlled conditions. The transgenic lines were fertile and did not exhibit any apparent morphological abnormality. The results of this study indicated an effective approach to improve the heavy metal accumulation ability of alfalfa plants which can then be used for the remediation of contaminated soil in arid regions.
Acknowledgments
We thank USDA, Agriculture Research Station, WRPIS for providing various genotypes of alfalfa for research purpose. We wish to thank Dr. MT Balba for providing oil-contaminated soil for the experiments. We would like to extend sincere thanks to Dr. Vittorio Venturi and Dr. Lawrence Banks for diligent proofreading of this paper. We thank Ms. Reshmy Sobha for contributing in statistical analysis.
Disclosure statement
No potential conflict of interest was reported by the authors.