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Activity of Arabidopsis Rubisco small subunit promoter in various tissues of chickpea

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Abstract

The activity of a green tissue-specific promoter of the Rubisco small subunit gene from Arabidopsis (AraSSU) was studied using transgenic chickpea lines. We generated transgenic chickpea lines expressing an AraSSU promoter-driven cry2Aa gene through the Agrobacterium-mediated transformation method. Lines with AraSSU expressed the gene in all green tissues at high levels (> 90 ng/mg of fresh weight tissue) compared to lines generated using CaMV35S (< 10 ng/mg FW). We used vertical cross sections of various tissues of homozygous progeny using microtome for immunolocalization. The immunolocalization showed the expression of the cry2Aa gene in the green mesophyll cells of the leaves of both AraSSU and CaMV35 chickpea lines. Moreover, the accumulation of AraSSU-regulated Cry2Aa protein was also observed in vascular tissues, including enucleate sieve elements and their companion cells. However, no expression was observed in the roots of AraSSU lines. In the case of CaMV35 lines, the transgene expression was observed in all the tissues. Since our data indicated that the AraSSU promoter is active in non-green tissues such as vascular bundles. Therefore, we validated this by RT-PCR. We found Cry2Aa RNA transcripts in leaves, stems without epidermis (for vascular tissues), and roots with and without epidermis. Thus, the AraSSU promoter is active in all above-ground tissues of the chickpea plant.

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Acknowledgements

We would like to express our thanks to Dr. P. Ananda Kumar (Indian Council of Agricultural Research-National Institute for Plant Biotechnology, New Delhi, India) for providing the pBin352A vector and Dr. Montu Bhuyan (Council of Scientific and Industrial Research-North East Institute of Science and Technology, Assam, India) for kindly providing the microtome used in this study. The authors acknowledge the Department of Biotechnology (DBT), Government of India, New Delhi, and the Indian Council of Agriculture Research (ICAR), New Delhi for funding assistance.

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Authors and Affiliations

  1. DBT-AAU Centre, Assam Agricultural University, Jorhat, Assam, 785013, India

    Rashmi Rekha Boruah, Trishna Konwar, Pranab Kumar Nath, Sumita Acharjee & Bidyut Kumar Sarmah

  2. Department of Agricultural Biotechnology, Assam Agricultural University, Jorhat, Assam, 785013, India

    Sumita Acharjee & Bidyut Kumar Sarmah

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  1. Rashmi Rekha Boruah
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  4. Sumita Acharjee
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Contributions

SA conceptualized and designed the experiments; BKS and SA contributed all the reagents, materials and analysis tools. RRB carried out transgenic experiments of CaMV35S-2Aa plants, RT-PCR analysis, ELISA, Immuno-histo-fluorescence, and statistical analysis. RRB, TK, SA, and BKS analyzed the data. RRB drafted the manuscript. SA edited the manuscript and supervised the work. BKS reviewed the manuscript. All the authors read and approved the manuscript.

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Correspondence to Sumita Acharjee or Bidyut Kumar Sarmah.

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Boruah, R.R., Konwar, T., Nath, P.K. et al. Activity of Arabidopsis Rubisco small subunit promoter in various tissues of chickpea. 3 Biotech 13, 89 (2023). https://doi.org/10.1007/s13205-023-03508-z

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