Skip to main content

Jasmonate Signaling in the Field, Part II: Insect-Guided Characterization of Genetic Variations in Jasmonate-Dependent Defenses of Transgenic and Natural Nicotiana attenuata Populations

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1011))

Abstract

The introduction of genetically modified plants into natural habitats represents a valuable means to determine organismic level functions of a gene and its effects on a plant’s interaction with other organisms. Nicotiana attenuata, a wild tobacco species native of the southwestern USA that grows in the immediate postfire environment, is one of the important host plants for herbivore populations recolonizing recently burned habitats in the Great Basin Desert. Here, we provide detailed guidelines for the analysis, under field conditions, of jasmonate-dependent defense and its impact on the plant’s native herbivore community. The procedures are based on the field release of transgenic lines silenced for jasmonate biogenesis, metabolism, or perception to conduct association studies between defense trait expression (secondary metabolite and trypsin proteinase inhibitor accumulation) and insect infestations. Additionally, because some insects have evolved mechanisms to “eavesdrop” on jasmonate signaling when selecting their host plants, we describe how leafhoppers of the species Empoasca, which selectively colonize jasmonate-deficient plants, can be used as “bloodhounds” for identifying natural variations in jasmonate signaling among natural N. attenuata populations.

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Pichersky E, Lewinsohn E (2011) Convergent evolution in plant specialized metabolism. Annu Rev Plant Biol 62:549–566

    Article  PubMed  CAS  Google Scholar 

  2. Schwachtje J, Baldwin IT (2004) Smoke exposure alters endogenous gibberellin and abscisic acid pools and gibberellin sensitivity while eliciting germination in the post-fire annual, Nicotiana attenuata. Seed Sci Res 14:51–60

    Article  CAS  Google Scholar 

  3. Baldwin IT, Staszak-Kozinski L, Davidson R (1994) Up in smoke: I. Smoke-derived germination cues for postfire annual, Nicotiana attenuata Torr. Ex. Watson. J Chem Ecol 20:2345–2371

    Article  CAS  Google Scholar 

  4. Baldwin IT, Morse L (1994) Up in smoke: II. Germination of Nicotiana attenuata in response to smoke-derived cues and nutrients in burned and unburned soils. J Chem Ecol 20:2373–2391

    Article  CAS  Google Scholar 

  5. Steppuhn A, Baldwin IT (2008) Induced defenses and the cost-benefit paradigm. In: Schaller A (ed) Induced plant resistance to herbivory. Springer, Berlin, pp 61–83

    Chapter  Google Scholar 

  6. Steppuhn A, Gase K, Krock B, Halitschke R, Baldwin IT (2004) Nicotine’s defensive function in nature. PLoS Biol 2:e217

    Article  PubMed  Google Scholar 

  7. Steppuhn A, Baldwin IT (2007) Resistance management in a native plant: nicotine prevents herbivores from compensating for plant protease inhibitors. Ecol Lett 10:499–511

    Article  PubMed  Google Scholar 

  8. Zavala JA, Patankar AG, Gase K, Hui D, Baldwin IT (2004) Manipulation of endogenous trypsin proteinase inhibitor production in Nicotiana attenuata demonstrates their function as antiherbivore defenses. Plant Physiol 134:1181–1190

    Article  PubMed  CAS  Google Scholar 

  9. Heiling S, Schuman M, Schöttner M, Mukerjee P, Berger B, Schneider B, Jassbi A, Baldwin IT (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

    Article  PubMed  CAS  Google Scholar 

  10. Kaur H, Heinzel N, Schöttner M, Baldwin IT, Gális I (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

    Article  PubMed  CAS  Google Scholar 

  11. Paschold A, Halitschke R, Baldwin IT (2007) Co(i)-ordinating defenses: NaCOI1 mediates herbivore-induced resistance in Nicotiana attenuata and reveals the role of herbivore movement in avoiding defenses. Plant J 51:79–91

    Article  PubMed  CAS  Google Scholar 

  12. Kessler A, Halitschke R, Baldwin IT (2004) Silencing the jasmonate cascade: induced plant defenses and insect populations. Science 305:665–668

    Article  PubMed  CAS  Google Scholar 

  13. Diezel C, Kessler D, Baldwin IT (2011) Pithy protection: Nicotiana attenuata’s jasmonic acid-mediated defenses are required to resist stem-boring weevil larvae. Plant Physiol 155:1936–1946

    Article  PubMed  CAS  Google Scholar 

  14. Bahulikar RA, Stanculescu D, Preston CA, Baldwin IT (2004) ISSR and AFLP analysis of the temporal and spatial population structure of the post-fire annual, Nicotiana attenuata, in SW Utah. BMC Ecol 4:12

    Article  PubMed  Google Scholar 

  15. Halitschke R, Schittko U, Pohnert G, Boland W, Baldwin IT (2001) Molecular interactions between the specialist herbivore Manduca sexta (Lepidoptera, Sphingidae) and its natural host Nicotiana attenuata. III. Fatty acid-amino acid conjugates in herbivore oral secretions are necessary and sufficient for herbivore-specific plant responses. Plant Physiol 125:711–717

    Article  PubMed  CAS  Google Scholar 

  16. Bonaventure G, VanDoorn A, Baldwin IT (2011) Herbivore-associated elicitors: FAC signaling and metabolism. Trends Plant Sci 16:294–299

    Article  PubMed  CAS  Google Scholar 

  17. McCloud ES, Baldwin IT (1997) Herbivory and caterpillar regurgitants amplify the wound-induced increases in jasmonic acid but not nicotine in Nicotiana sylvestris. Planta 203:430–435

    Article  CAS  Google Scholar 

  18. Kallenbach M, Bonaventure G, Gilardoni PA, Wissgott A, Baldwin IT (2012) Empoasca leafhoppers attack wild tobacco plants in a jasmonate-dependent manner and identify jasmonate mutants in natural populations. Proc Natl Acad Sci USA 109:1548–1557

    Article  Google Scholar 

  19. Stitz M, Gase K, Baldwin IT, Gaquerel E (2011) Ectopic expression of AtJMT in Nicotiana attenuata: creating a metabolic sink has tissue-specific consequences for the jasmonate metabolic network and silences downstream gene expression. Plant Physiol 157:341–354

    Article  PubMed  CAS  Google Scholar 

  20. Keinänen M, Oldham NJ, Baldwin IT (2001) Rapid HPLC screening of jasmonate-induced increases in tobacco alkaloids, phenolics, and diterpene glycosides in Nicotiana attenuata. J Agric Food Chem 49:3553–3558

    Article  PubMed  Google Scholar 

  21. Gaquerel E, Heiling S, Schoettner M, Zurek G, Baldwin IT (2010) Development and validation of a liquid chromatography-electrospray ionization-time-of-flight mass spectrometry method for induced changes in Nicotiana attenuata leaves during simulated herbivory. J Agric Food Chem 58:9418–9427

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Gaquerel, E., Stitz, M., Kallenbach, M., Baldwin, I.T. (2013). Jasmonate Signaling in the Field, Part II: Insect-Guided Characterization of Genetic Variations in Jasmonate-Dependent Defenses of Transgenic and Natural Nicotiana attenuata Populations. In: Goossens, A., Pauwels, L. (eds) Jasmonate Signaling. Methods in Molecular Biology, vol 1011. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-414-2_8

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-414-2_8

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-413-5

  • Online ISBN: 978-1-62703-414-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics