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Virus-Induced Gene Silencing in Cultivated Cotton (Gossypium spp.) Using Tobacco Rattle Virus

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Abstract

The study described here has optimized the conditions for virus-induced gene silencing (VIGS) in three cultivated cotton species (Gossypium hirsutum, G. arboreum, and G. herbaceum) using a Tobacco rattle virus (TRV) vector. The system was used to silence the homolog of the Arabidopsis thaliana chloroplastos alterados 1 (AtCLA1) gene, involved in chloroplast development, in G. herbaceum, G. arboreum, and six commercial G. hirsutum cultivars. All plants inoculated with the TRV vector to silence CLA1 developed a typical albino phenotype indicative of silencing this gene. Although silencing in G. herbaceum and G. arboreum was complete, silencing efficiency differed for each G. hirsutum cultivar. Reverse transcriptase polymerase chain reaction (PCR) and real-time quantitative PCR showed a reduction in mRNA levels of the CLA1 homolog in all three species, with the highest efficiency (lowest CLA1 mRNA levels) in G. arboreum followed by G. herbaceum and G. hirsutum. The results indicate that TRV is a useful vector for VIGS in Gossypium species. However, selection of host cultivar is important. With the genome sequences of several cotton species recently becoming publicly available, this system has the potential to provide a very powerful tool for the rapid, large-scale reverse-genetic analysis of genes in Gossypium spp.

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Acknowledgments

The authors are grateful to Dr. Ping He (Department of Biochemistry and Biophysics, Institute of Plant Genomics and Biotechnology, Texas A&M University, Texas, USA) for providing the TRV-GrCLA1 construct and Dr. Mehboob-ur-Rehman (NIBGE, Faisalabad) for providing cotton seeds. This material is based upon work supported by the “Pak-US cotton productivity enhancement program” of the International Center for Agricultural Research in the Dry Areas (ICARDA) funded by United States Department of Agriculture (USDA), Agricultural Research Service (ARS), under Agreement No. 58-6402-0-178F. Any opinions, findings, conclusions or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the views of the USDA or ICARDA.

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Authors and Affiliations

  1. Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan

    Roma Mustafa, Muhammad Shafiq, Shahid Mansoor, Rob W. Briddon & Imran Amin

  2. Genomics and Bioinformatics Research Unit, PO Box 36, Stoneville, MS, 38776, USA

    Brian E. Scheffler & Jodi Scheffler

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  1. Roma Mustafa
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  2. Muhammad Shafiq
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  3. Shahid Mansoor
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  4. Rob W. Briddon
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  5. Brian E. Scheffler
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  6. Jodi Scheffler
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  7. Imran Amin
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Correspondence to Imran Amin.

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Mustafa, R., Shafiq, M., Mansoor, S. et al. Virus-Induced Gene Silencing in Cultivated Cotton (Gossypium spp.) Using Tobacco Rattle Virus . Mol Biotechnol 58, 65–72 (2016). https://doi.org/10.1007/s12033-015-9904-z

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  • DOI: https://doi.org/10.1007/s12033-015-9904-z

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