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Chilli leaf curl virus-based vector for phloem-specific silencing of endogenous genes and overexpression of foreign genes

Applied Microbiology and Biotechnology volume 101, pages 2121–2129 (2017)Cite this article

Abstract

Geminiviruses are the largest and most devastating group of plant viruses which contain ssDNA as a genetic material. Geminivirus-derived virus-induced gene silencing (VIGS) vectors have emerged as an efficient and simple tool to study functional genomics in various plants. However, previously developed VIGS vectors have certain limitations, owing to their inability to be used in tissue-specific functional study. In the present study, we developed a Chilli leaf curl virus (ChiLCV)-based VIGS vector for its tissue-specific utilization by replacing the coat protein gene (open reading frame (ORF) AV1) with the gene of interest for phytoene desaturase (PDS) of Nicotiana benthamiana. Functional validation of ChiLCV-based VIGS in N. benthamiana resulted in systemic silencing of PDS exclusively in the phloem region of inoculated plants. Furthermore, expression of enhanced green fluorescence protein (EGFP) using the same ChiLCV vector was verified in the phloem region of the inoculated plants. Our results also suggested that, during the early phase of infection, ChiLCV was associated with the phloem region, but at later stage of pathogenesis, it can spread into the adjoining non-vascular tissues. Taken together, the newly developed ChiLCV-based vector provides an efficient and versatile tool, which can be exploited to unveil the unknown functions of several phloem-specific genes.

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Acknowledgments

The authors thank Mr. Pranav Pankaj Sahu for his critical comments and suggestions on the manuscript.

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Affiliations

  1. Molecular Virology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110 067, India

    Nirbhay Kumar Kushwaha & Supriya Chakraborty

Authors
  1. Nirbhay Kumar Kushwaha
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  2. Supriya Chakraborty
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Correspondence to Supriya Chakraborty.

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Funding

This study was funded by the University Grant Commission Resource Net Working programme (UGC-RNW/SLS/SC) and the Department of Science and Technology, Ministry of Science and Technology, India (DST-PURSE/JNU/SC).

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The authors declare that they have no conflict of interest.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Kushwaha, N.K., Chakraborty, S. Chilli leaf curl virus-based vector for phloem-specific silencing of endogenous genes and overexpression of foreign genes. Appl Microbiol Biotechnol 101, 2121–2129 (2017). https://doi.org/10.1007/s00253-016-7964-z

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  • DOI: https://doi.org/10.1007/s00253-016-7964-z

Keywords

  • ChiLCV
  • Geminivirus
  • VIGS vector
  • Phloem specific
  • Virus movement
  • Expression vector