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A Method for Developing RNAi-Derived Resistance in Cowpea Against Geminiviruses

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Plant Gene Silencing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2408))

Abstract

In plants, RNA interference (RNAi) is triggered by double-stranded RNA (dsRNA). Accordingly, various RNA silencing technologies involving hpRNA, artificial microRNA (miRNA), and virus-induced gene silencing (VIGS) are used for controlling the expression of genes. Such manipulations help understanding gene functions and crop improvement biotechnology. A typical hpRNA construct is comprised of an intron splicable perfect inverted repeat of the target gene sequences under the control of a strong promoter. Geminiviruses, especially Mungbean Yellow Mosaic India Virus (MYMIV) cause devastating diseases in legume plants including cowpea, incurring severe crop loss. RNAi, involving hpRNA construct as transgene, is used to control these diseases at the early stages of geminivirus infection in the host, preventing symptom development and viral DNA accumulation. In this chapter, we describe a detailed protocol for the identification of geminivirus isolates from the filed grown cowpea plants, characterization of virus isolates under the laboratory conditions, design and construct RNAi vectors for effective suppression of viral target genes, and consequent development of transgenic cowpea using Agrobacterium-mediated transformation protocol. These transgenics are subsequently evaluated for resistance to MYMIV.

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Author information

Authors and Affiliations

  1. Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, India

    Sanjeev Kumar & Lingaraj Sahoo

  2. Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi, India

    Sunil Kumar Mukherjee

Authors
  1. Sanjeev Kumar
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  2. Sunil Kumar Mukherjee
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  3. Lingaraj Sahoo
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Corresponding author

Correspondence to Lingaraj Sahoo .

Editor information

Editors and Affiliations

  1. Institute for Agricultural Biosciences, Oklahoma State University, Ardmore, OK, USA

    Kirankumar S. Mysore

  2. National Institute of Plant Genome Research, New Delhi, India

    Muthappa Senthil-Kumar

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Kumar, S., Mukherjee, S.K., Sahoo, L. (2022). A Method for Developing RNAi-Derived Resistance in Cowpea Against Geminiviruses. In: Mysore, K.S., Senthil-Kumar, M. (eds) Plant Gene Silencing. Methods in Molecular Biology, vol 2408. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1875-2_13

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  • DOI: https://doi.org/10.1007/978-1-0716-1875-2_13

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1874-5

  • Online ISBN: 978-1-0716-1875-2

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