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Targeted Metabolomics of Plant Hormones and Redox Metabolites in Stomatal Immunity

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Jasmonate in Plant Biology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2085))

Abstract

Phytohormones and redox metabolites are important molecules in a number of biological processes related to plant growth, development, and stress responses. Understanding how these metabolites are involved in abiotic and biotic stress is a frequent topic of plant biology research. However, many factors, such as low physiological concentrations and the inherent complexity of plant samples, make identification and quantification of these important metabolites difficult. Here, we describe a method for metabolite extraction from whole leaves and guard cell–enriched samples and a targeted metabolomics strategy for the identification and quantification of specific hormone- and redox-related metabolites. In our experiment, we used the reference plant Arabidopsis thaliana infected with the biotrophic pathogen Pseudomonas syringe pv. tomato (Pst) DC3000, and examined the changes in hormone and redox metabolites in systemic leaves, using the targeted metabolomics strategy in order to investigate potential functions of these metabolites in systemic acquired resistance (SAR) during a plant’s immune responses. The methods reported here can be expanded to other metabolites and other biological systems beyond plants and bacterial pathogens.

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References

  1. Verma V, Ravindran P, Kumar PP (2016) Plant hormone-mediated regulation of stress responses. BMC Plant Biol 16:86. https://doi.org/10.1186/s12870-016-0771-y

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Nguyen D, Rieu I, Mariani C, van Dam NM (2016) How plants handle multiple stresses: hormonal interactions underlying responses to abiotic stress and insect herbivory. Plant Mol Biol 91:727–740. https://doi.org/10.1007/s11103-016-0481-8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Fan J, Hill L, Crooks C, Doerner P, Lamb C (2009) Abscisic acid has a key role in modulating diverse plant-pathogen interactions. Plant Physiology 150:1750–1761. https://doi.org/10.1104/pp.109.137943

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Van Breusegem F, Bailey-Serres J, Mittler R (2008) Unraveling the tapestry of networks involving reactive oxygen species in plants. Plant Physiol 147:978–984. https://doi.org/10.1104/pp.108.122325

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Misra BB, Chaturvedi R (2015) When plants brace for the emerging pathogens. Physiol Mol Plant Pathol 92:181–185. https://doi.org/10.1016/j.pmpp.2015.03.004

    Article  Google Scholar 

  6. Spoel SH, Dong XN (2012) How do plants achieve immunity? Defense without specialized immune cells. Nat Rev Immunol 12(2):89–100. https://doi.org/10.1038/nri3141

    Article  CAS  PubMed  Google Scholar 

  7. Fu ZQ, Dong XN (2013) Systemic acquired resistance: turning local infection into global defense. Annu Rev Plant Biol 64:839–863. https://doi.org/10.1146/annurev-arplant-042811-105606

    Article  CAS  PubMed  Google Scholar 

  8. Park SW, Kaimoyo E, Kumar D, Mosher S, Klessig DF (2007) Methyl salicylate is a critical mobile signal for plant systemic acquired resistance. Science 318(5847):113–116. https://doi.org/10.1126/science.1147113

    Article  CAS  PubMed  Google Scholar 

  9. Kachroo A, Robin GP (2013) Systemic signaling during plant defense. Curr Opin Plant Biol 16:527–533. https://doi.org/10.1016/j.pbi.2013.06.019

    Article  CAS  PubMed  Google Scholar 

  10. Sels J, Mathys J, De Coninck BMA, Cammue BPA, De Bolle MFC (2008) Plant pathogenesis-related (PR) proteins: a focus on PR peptides. Plant Physiol Biochem 46(11):941–950. https://doi.org/10.1016/j.plaphy.2008.06.011

    Article  CAS  Google Scholar 

  11. Shah J, Zeier J (2013) Long-distance communication and signal amplification in systemic acquired resistance. Front Plant Sci 4(30):1–16. https://doi.org/10.3389/fpls.2013.00030

    Article  CAS  Google Scholar 

  12. Dempsey DA, Klessig DF (2012) SOS – too many signals for systemic acquired resistance? Trends Plant Sci 17(9):538–545. https://doi.org/10.1016/j.tplants.2012.05.011

    Article  CAS  PubMed  Google Scholar 

  13. Zeng W, Brutus A, Kremer JM, Withers JC, Gao X, Jones AD et al (2011) A genetic screen reveals arabidopsis stomatal and/or apoplastic defenses against Pseudomonas syringae pv. tomato DC3000. PLoS Pathol 7:10. https://doi.org/10.1371/journal.ppat.1002291

    Article  CAS  Google Scholar 

  14. Melotto M, Underwood W, He SY (2008) Role of stomata in plant innate immunity and foliar bacterial diseases. Annu Rev Phytopathol 46:101–122. https://doi.org/10.1146/annurev.phyto.121107.104959

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. McLachlan DH, Kopischke M, Robatzek S (2014) Gate control: guard cell regulation by microbial stress. New Phytol 203:1049–1063. https://doi.org/10.1111/nph.12916

    Article  CAS  PubMed  Google Scholar 

  16. Sawinski K, Mersmann S, Robatzek S, Bohmer M (2013) Guarding the green: pathways to stomatal immunity. Mol Plant-Microbe Interact 26:626–632. https://doi.org/10.1094/MPMI-12-12-0288-CR

    Article  CAS  PubMed  Google Scholar 

  17. Melotto M, Underwood W, Koczan J, Nomura K, He SY (2006) Plant stomata function in innate immunity against bacterial invasion. Cell 126(5):969–980. https://doi.org/10.1016/j.cell.2006.06.054

    Article  CAS  PubMed  Google Scholar 

  18. Assmann SM, Jegla T (2016) Guard cell sensory systems: recent insights on stomatal responses to light, abscisic acid, and CO2. Curr Opin Plant Biol 33:157–167. https://doi.org/10.1016/j.pbi.2016.07.003

    Article  CAS  PubMed  Google Scholar 

  19. Shah J, Chaturvedi R, Chowdhury Z, Venables B, Petros RA (2014) Signaling by small metabolites in systemic acquired resistance. Plant J 79(4):645–658. https://doi.org/10.1111/tpj.12464

    Article  CAS  PubMed  Google Scholar 

  20. Bauer H, Ache P, Lautner S, Fromm J, Hartung W, Al-Rasheid KA, Sonnewald S, Sonnewald U, Kneitz S, Lachmann N, Mendel RR, Bittner F, Hetherington AM, Hedrich R (2013) The stomatal response to reduced relative humidity requires guard cell-autonomous ABA synthesis. Curr Biol 23:53–57. https://doi.org/10.1016/j.cub.2012.11.022

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Sixue Chen .

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David, L., Kang, J., Chen, S. (2020). Targeted Metabolomics of Plant Hormones and Redox Metabolites in Stomatal Immunity. In: Champion, A., Laplaze, L. (eds) Jasmonate in Plant Biology. Methods in Molecular Biology, vol 2085. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0142-6_6

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  • DOI: https://doi.org/10.1007/978-1-0716-0142-6_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0141-9

  • Online ISBN: 978-1-0716-0142-6

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