Abstract
Plants employ a large variety of defense strategies to resist herbivores, which require transcriptional reprogramming of cells and profound changes in plant metabolism. Due to the large number of genes involved in defense processes, rapid screening strategies are essential for elucidating the contributions of individual genes in the responses of plants to herbivory. However, databases and seed banks of mutant plants which allow rapid retrieval of mutant genotypes are limited to a few model plant species, namely, Arabidopsis thaliana and Oryza sativa (rice). In other plants, virus-induced gene silencing (VIGS) offers an efficient alternative for screening the functions of individual genes in order to prioritize the allocations of the large time investments required to establish stably transformed RNAi-silenced lines. With VIGS, it is usually possible to achieve strong, specific silencing of target genes in the ecological models Nicotiana attenuata and Solanum nigrum, allowing the rapid assessment of gene silencing effects on phytohormone accumulation, signal transduction and accumulation of defense metabolites. VIGS plants are also useful in bioassays with specialist and generalist herbivores, allowing direct verification of gene function in plant resistance to herbivores.
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References
Ratcliff F, Martin-Hernandez AM, Baulcombe DC (2001) Tobacco rattle virus as a vector for analysis of gene function by silencing. Plant J 25:237–245
Kessler A, Halitschke R, Baldwin IT (2004) Silencing the jasmonate cascade: induced plant defenses and insect populations. Science 305:665–668
Steppuhn A, Gase K, Krock B et al (2004) Nicotine’s defensive function in nature. PLoS Biol 2:e217
Zavala JA, Patankar AG, Gase K et al (2004) Constitutive and inducible trypsin proteinase inhibitor production incurs large fitness costs in Nicotiana attenuata. Proc Natl Acad Sci USA 101:1607–1612
Kang JH, Wang L, Giri A et al (2006) Silencing threonine deaminase and JAR4 in Nicotiana attenuata impairs jasmonic acid-isoleucine-mediated defenses against Manduca sexta. Plant Cell 18:3303–3320
Schwachtje J, Minchin PEH, Jahnke S et al (2006) SNF1-related kinases allow plants to tolerate herbivory by allocating carbon to roots. Proc Natl Acad Sci USA 103:12935–12940
Wu J, Hettenhausen C, Meldau S et al (2007) Herbivory rapidly activates MAPK signaling in attacked and unattacked leaf regions but not between leaves of Nicotiana attenuata. Plant Cell 19:1096–1122
Jassbi AR, Gase K, Hettenhausen C et al (2008) Silencing geranylgeranyldiphosphate synthase in Nicotiana attenuata dramatically impairs resistance to tobacco hornworm. Plant Physiol 146:974–986
Kessler D, Gase K, Baldwin IT (2008) Field experiments with transformed plants reveal the sense of floral scents. Science 321:1200–1202
Skibbe M, Qu N, Gális I et al (2008) Induced plant defenses in the natural environment: Nicotiana attenuata WRKY3 and WRKY6 coordinate responses to herbivory. Plant Cell 20:1984–2000
Heiling S, Schuman M, Schöttner M et al (2010) Jasmonate and ppHsystemin regulate key malonylation steps in the biosynthesis of 17-hydroxygeranyllinalool diterpene glycosides, an abundant and effective direct defense against herbivores in Nicotiana attenuata. Plant Cell 22:273–292
Kaur H, Heinzel N, Schöttner M et al (2010) R2R3-NaMYB8 regulates the accumulation of phenylpropanoid-polyamine conjugates, which are essential for local and systemic defense against insect herbivores in Nicotiana attenuata. Plant Physiol 152:1731–1747
Hartl M, Giri A, Kaur H et al (2010) Serine protease inhibitors specifically defend Solanum nigrum against generalist herbivores but do not influence plant growth and development. Plant Cell 22:4158–4175
Hartl M, Merker H, Schmidt DD et al (2008) Optimized virus-induced gene silencing in Solanum nigrum reveals the defensive function of leucine aminopeptidase against herbivores and the shortcomings of empty vector controls. New Phytol 179:356–365
Liu Y, Schiff M, Marathe R et al (2002) Tobacco Rar1, EDS1 and NPR1/NIM1 like genes are required for N-mediated resistance to tobacco mosaic virus. Plant J 30:415–429
Ploeg AT, Robinson DJ, Brown DJF (1993) RNA-2 of tobacco rattle virus encodes the determinants of transmissibility by trichodorid vector nematodes. J Gen Virol 74:1463–1466
Hamilton RI (1974) Replication of plant viruses. Annu Rev Phytopathol 12:223–245
Voinnet O (2005) Induction and suppression of RNA silencing: insights from viral infections. Nat Rev Genet 6:206–U201
Saedler R, Baldwin IT (2004) Virus-induced gene silencing of jasmonate-induced direct defences, nicotine and trypsin proteinase-inhibitors in Nicotiana attenuata. J Exp Bot 55:151–157
Schittko U, Preston CA, Baldwin IT (2000) Eating the evidence? Manduca sexta larvae can not disrupt specific jasmonate induction in Nicotiana attenuata by rapid consumption. Planta 210:343–346
Brigneti G, Martin-Hernandez AM, Jin H et al (2004) Virus-induced gene silencing in Solanum species. Plant J 39:264–272
Wu C, Jia L, Goggin F (2011) The reliability of virus-induced gene silencing experiments using tobacco rattle virus in tomato is influenced by the size of the vector control. Mol Plant Pathol 12:299–305
Thomas CL, Jones L, Baulcombe DC et al (2001) Size constraints for targeting post-transcriptional gene silencing and for RNA-directed methylation in Nicotiana benthamiana using a potato virus X vector. Plant J 25:417–425
Schmidt DD, Kessler A, Kessler D et al (2004) Solanum nigrum: a model ecological expression system and its tools. Mol Ecol 13:981–995
Gachon C, Mingam A, Charrier B (2004) Real-time PCR: what relevance to plant studies? J Exp Bot 55:1445–1454
Deepak S, Kottapalli K, Rakwal R et al (2007) Real-time PCR: revolutionizing detection and expression analysis of genes. Curr Genomics 8:234–251
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Galis, I. et al. (2013). The Use of VIGS Technology to Study Plant–Herbivore Interactions. In: Becker, A. (eds) Virus-Induced Gene Silencing. Methods in Molecular Biology, vol 975. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-278-0_9
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DOI: https://doi.org/10.1007/978-1-62703-278-0_9
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