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Virus-Induced Gene Silencing in Nepeta

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Virus-Induced Gene Silencing in Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2172))

Abstract

Virus-induced gene silencing (VIGS) is a versatile tool for genetic studies that has been applied to a variety of plant species. With the advent of more accessible genomic and transcriptomic technology applied to an increasing range of plants, tools such as VIGS are being adapted to more non-model plants to explore genes relevant to agriculture and chemical discovery. In this protocol, we adapted VIGS technology to target genes in Nepeta cataria (catnip) and Nepeta mussinii (catmint). These plants carry biochemical and economical value for their production of nepetalactone, an iridoid which provokes a strong reaction in both house cats and aphids. We describe a method to target magnesium chelatase subunit H (CHlH), a gene often targeted as a visual marker for VIGS. Furthermore, we describe a method to simultaneously target two genes in a single plant, which aids in the study of genes found in key biochemical steps in the production of nepetalactone. This approach, which was successfully applied in two members of the Lamiaceae family (mint), could be adapted to other members of the mint family with economical and chemical value.

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References

  1. Lange M, Yellina AL, Orashakova S, Becker A (2013) Virus-Induced Gene Silencing (VIGS) in plants: an overview of target species and the virus-derived vector systems. In: Becker A (ed) Virus-induced gene silencing: methods and protocols. Humana Press, Totowa, NJ, pp 1–14. https://doi.org/10.1007/978-1-62703-278-0_1

    Chapter  Google Scholar 

  2. Robertson D (2004) VIGS vectors for gene silencing: many targets, many tools. Annu Rev Plant Biol 55(1):495–519. https://doi.org/10.1146/annurev.arplant.55.031903.141803

    Article  CAS  PubMed  Google Scholar 

  3. Liu Y, Schiff M, Dinesh-Kumar SP (2002) Virus-induced gene silencing in tomato. Plant J 31(6):777–786. https://doi.org/10.1046/j.1365-313X.2002.01394.x

    Article  CAS  PubMed  Google Scholar 

  4. Liu Y, Schiff M, Marathe R, Dinesh-Kumar SP (2002) Tobacco Rar1, EDS1 and NPR1/NIM1 like genes are required for N-mediated resistance to tobacco mosaic virus. Plant J 30(4):415–429. https://doi.org/10.1046/j.1365-313X.2002.01297.x

    Article  CAS  PubMed  Google Scholar 

  5. Velásquez AC, Chakravarthy S, Martin GB (2009) Virus-induced gene silencing (VIGS) in Nicotiana benthamiana and tomato. J Vis Exp (28):1292. https://doi.org/10.3791/1292

  6. Liu E, Page JE (2008) Optimized cDNA libraries for virus-induced gene silencing (VIGS) using tobacco rattle virus. Plant Methods 4(1):5. https://doi.org/10.1186/1746-4811-4-5

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Golenberg EM, Sather DN, Hancock LC, Buckley KJ, Villafranco NM, Bisaro DM (2009) Development of a gene silencing DNA vector derived from a broad host range geminivirus. Plant Methods 5:9. https://doi.org/10.1186/1746-4811-5-9

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Lu R, Peart JR, Malcuit I, Baulcombe DC (2003) Virus-induced gene silencing in plants. Methods 30:296–303. https://doi.org/10.1016/S1046-2023(03)00037-9

    Article  CAS  PubMed  Google Scholar 

  9. Geu-Flores F, Sherden NH, Courdavault V, Burlat V, Glenn WS, Wu C et al (2012) An alternative route to cyclic terpenes by reductive cyclization in iridoid biosynthesis. Nature 492:138. https://doi.org/10.1038/nature11692

    Article  CAS  PubMed  Google Scholar 

  10. Boachon B, Buell CR, Crisovan E, Dudareva N, Garcia N, Godden G et al (2018) Phylogenomic mining of the mints reveals multiple mechanisms contributing to the evolution of chemical diversity in Lamiaceae. Mol Plant 11(8):1084–1096. https://doi.org/10.1016/j.molp.2018.06.002

    Article  CAS  Google Scholar 

  11. Weng J-K, Philippe RN, Noel JP (2012) The rise of chemodiversity in plants. Science 336(6089):1667–1670. https://doi.org/10.1126/science.1217411

    Article  CAS  PubMed  Google Scholar 

  12. Lichman BR, Kamileen MO, Titchiner GR, Saalbach G, Stevenson CEM, Lawson DM, O’Connor SE (2018) Uncoupled activation and cyclization in catmint reductive terpenoid biosynthesis. Nat Chem Biol 15(1):71–79. https://doi.org/10.1101/391953

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Misra RC, Sharma S, Sandeep, Garg A, Chanotiya CS, Ghosh S (2017) Two CYP716A subfamily cytochrome P450 monooxygenases of sweet basil play similar but nonredundant roles in ursane- and oleanane-type pentacyclic triterpene biosynthesis. New Phytol 214(2):706–720. https://doi.org/10.1111/nph.14412

    Article  CAS  PubMed  Google Scholar 

  14. Chakraborty P (2018) Herbal genomics as tools for dissecting new metabolic pathways of unexplored medicinal plants and drug discovery. Biochim Open 6:9–16. https://doi.org/10.1016/j.biopen.2017.12.003

    Article  PubMed  PubMed Central  Google Scholar 

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Correspondence to Sarah E. O’Connor .

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Palmer, L., O’Connor, S.E. (2020). Virus-Induced Gene Silencing in Nepeta. In: Courdavault, V., Besseau, S. (eds) Virus-Induced Gene Silencing in Plants. Methods in Molecular Biology, vol 2172. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0751-0_9

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  • DOI: https://doi.org/10.1007/978-1-0716-0751-0_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0750-3

  • Online ISBN: 978-1-0716-0751-0

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