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High Efficiency Transformation of Banana [Musa acuminata L. cv. Matti (AA)] for Enhanced Tolerance to Salt and Drought Stress Through Overexpression of a Peanut Salinity-Induced Pathogenesis-Related Class 10 Protein

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Abstract

Bananas and plantains (Musa spp. L.) are important subsistence crops and premium export commodity in several countries, and susceptible to a wide range of environmental and biotic stress conditions. Here, we report efficient, rapid, and reproducible Agrobacterium-mediated transformation and regeneration of an Indian niche cultivar of banana [M. acuminata cv. Matti (AA)]. Apical meristem-derived highly proliferative multiple shoot clump (MSC) explants were transformed with the Agrobacterium strain EHA105 harboring a binary vector pCAMBIA-1301 carrying hptII and uidA. Sequential agro-infiltration (10 min, 400 mmHg), infection (additional 35 min, Agrobacterium density A 600 = 0.8) and co-cultivation (18 h) regimen in 100 µM acetosyringone containing liquid medium were critical factors yielding high transformation efficiency (~81 %) corroborated by transient GUS expression assay. Stable transgenic events were recovered following two cycles of meristem initiation and selection on hygromycin containing medium. Histochemical GUS assay in several tissues of transgenic plants and molecular analyses confirmed stable integration and expression of transgene. The protocol described here allowed recovery of well-established putative transgenic plantlets in as little as 5 months. The transgenic banana plants could be readily acclimatized under greenhouse conditions, and were phenotypically similar to the wild-type untransformed control plants (WT). Transgenic plants overexpressing Salinity-Induced Pathogenesis-Related class 10 protein gene from Arachis hypogaea (AhSIPR10) in banana cv. Matti (AA) showed better photosynthetic efficiency and less membrane damage (P < 0.05) in the presence of NaCl and mannitol in comparison to WT plants suggesting the role of AhSIPR10 in better tolerance of salt stress and drought conditions.

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Abbreviations

AhSIPR10 :

Arachis hypogaea salinity-induced PR class 10 gene

ECS:

Embryogenic cell suspension

MS:

Murashige and Skoog medium

MSC:

Multiple shoot clump

PR10:

Pathogenesis-related (class 10)

WT:

Wild-type

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Acknowledgments

This research work was supported by Department of Biotechnology, India (Grant no BT/PR10231/AGR/02/555/2007) to NBS. A.R. acknowledges the financial support from University Grant Commission (UGC). The authors also wish to thank Dr. Anuradha Agarwal, and Dr. Kailash Chander Bansal, Director, National Bureau of Plant Genetic Resources, New Delhi, India for disease-free banana plants. Research in the laboratory of NBS is supported by U.G.C.-C.A.S., U.G.C.-R.N.W., Department of Science and Technology (D.S.T.)-F.I.S.T., and D.S.T.-PURSE.

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Author notes

  1. Anjana Rustagi

    Present address: Department of Botany, Ramjas College, University Enclave, New Delhi, 110007, India

  2. Shalu Jain

    Present address: Department of Plant Pathology, North Dakota State University, Fargo, ND, 58102, USA

  3. Mukesh Jain

    Present address: Department of Plant Pathology, University of Florida, Gainesville, FL, 32610, USA

  4. Vishnu Bhat

    Present address: Department of Botany, University of Delhi, Delhi, 110007, India

Authors and Affiliations

  1. Plant Developmental Biology and Transformation Lab, School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110067, India

    Anjana Rustagi, Shalu Jain, Deepak Kumar, Shashi Shekhar, Mukesh Jain & Neera Bhalla Sarin

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  1. Anjana Rustagi
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Correspondence to Neera Bhalla Sarin.

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Rustagi, A., Jain, S., Kumar, D. et al. High Efficiency Transformation of Banana [Musa acuminata L. cv. Matti (AA)] for Enhanced Tolerance to Salt and Drought Stress Through Overexpression of a Peanut Salinity-Induced Pathogenesis-Related Class 10 Protein. Mol Biotechnol 57, 27–35 (2015). https://doi.org/10.1007/s12033-014-9798-1

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