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Transient Expression Analysis of Agave sisalana Heat Shock Protein Gene (AsHSP70) in Model Species (Nicotiana benthamiana) under Heat Stress

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Abstract

This study is aimed at investigating whether overexpression of the heat shock protein gene (AsHSP70) isolated from Agave sisalana will improve the heat stress tolerance in plants. AsHSP70 gene with a full-length cDNA of 1800 bp, encoding 282 amino acids, was functionally characterized. Bioinformatics tools showed the theoretical isoelectric point is pI = 4.780, located in the nucleus with 76.8% reliability having an N-terminal signal domain. 3D model, showed an ATP and ADP ligands with –6.2 and 6.5 kJ/mol binding affinity respectively. AsHSP70 was cloned in Plant expression vector in fusion with Green Florescent Protein from pGWB5 vector. Syringe agroinfiltration of Agrobacterium tumefaciens harboring AsHSP70/GFP was done into leaf of Nicotiana benthamiana. Spermidine was applied to the infiltrated tobacco plants for exogenous induction of polyamines under heat stress. Relative expression of AsHSP70 was significant in tobacco leaf under heat stress. Membrane stability index was about 95.59 and 96.53% and leaf relative water content was 4.43 and 3.44% respectively in infiltrated and polyamine-induced plants. Proline and sugar content were also increased under heat stress. Hence, we established that AsHSP70 is expressing transiently under heat stress in tobacco as model species and further genetic transformation of this gene will lead to induce the stress tolerance in agriculturally important crops.

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ACKNOWLEDGMENTS

The authors would like to acknowledge the Higher Education Commission Pakistan (HEC) and The World Academy of Science (TWAS) for provision of funds to complete this study.

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Authors and Affiliations

  1. Centre of Excellence in Molecular Biology, University of the Punjab Lahore, 53700, Lahore, Pakistan

    A. A. Batcho, B. Jabbar, M. B. Sarwar, B. Rashid, S. Hassan & T. Husnain

  2. Catholic University of the West Africa Benin, Cotonou, Benin

    A. A. Batcho

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  1. A. A. Batcho
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  2. B. Jabbar
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  3. M. B. Sarwar
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  4. B. Rashid
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  5. S. Hassan
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  6. T. Husnain
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Contributions

This study has been designed by TH and managed by BR. The overall experiments have been done by BAA, Bioinformatics analyses has been performed by BJ, Real time qPCR has been done by SMB. Statistical and graphical data presentation was done by SH. Manuscript drafted by BR and TH edited the final version. All the authors have read and approved the manuscript.

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Correspondence to B. Rashid.

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The authors declare that they have no conflicts of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.

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Batcho, A.A., Jabbar, B., Sarwar, M.B. et al. Transient Expression Analysis of Agave sisalana Heat Shock Protein Gene (AsHSP70) in Model Species (Nicotiana benthamiana) under Heat Stress. Biol Bull Russ Acad Sci 49, 160–168 (2022). https://doi.org/10.1134/S1062359022030037

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