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Investigation of Auxin Biosynthesis and Action Using Auxin Biosynthesis Inhibitors

Part of the Methods in Molecular Biology book series (MIMB,volume 2213)


Auxin plays important roles in almost all aspects of plant growth and development. Chemical genetics is an effective approach to understand auxin action, especially in nonmodel plant species, in which auxin-related mutants are not yet available. Among auxin-related chemical tools, we present approaches to utilize auxin biosynthesis inhibitors. The inhibitors are effective not only to understand auxin biosynthesis but also to understand auxin action. The effectiveness of the inhibitors can be assessed based on in vitro or in vivo assays. The in vitro assay employs enzyme inhibition assays. The in vivo assay employs UPLC-MS/MS-based analysis of endogenous IAA and its intermediates or metabolites.

Key words

  • Chemical genetics
  • Arabidopsis
  • Auxin biosynthesis
  • Reverse genetics

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  • DOI: 10.1007/978-1-0716-0954-5_12
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  1. De Rybel B, Audenaert D, Beeckman T, Kepinski S (2009) The past, present, and future of chemical biology in auxin research. ACS Chem Biol 4:987–998.

  2. Ma Q, Robert S (2014) Auxin biology revealed by small molecules. Physiol Plant 151:25–42.

  3. Hayashi K, Joshua N, Hirose M et al (2012) Rational design of an Auxin antagonist of the SCF TIR1 auxin receptor complex. ACS Chem Biol 7:590–598.

  4. Dhonukshe P, Grigoriev I, Fischer R et al (2008) Auxin transport inhibitors impair vesicle motility and actin cytoskeleton dynamics in diverse eukaryotes. Proc Natl Acad Sci U S A 105:4489–4494.

  5. Zhu J, Bailly A, Zwiewka M et al (2016) TWISTED DWARF1 mediates the action of auxin transport inhibitors on actin cytoskeleton dynamics. Plant Cell 28:930–948.

  6. Stepanova AN, Robertson-Hoyt J, Yun J et al (2008) TAA1-mediated auxin biosynthesis is essential for hormone crosstalk and plant development. Cell 133:177–191.

  7. Tao Y, Ferrer J-L, Ljung K et al (2008) Rapid synthesis of auxin via a new tryptophan-dependent pathway is required for shade avoidance in plants. Cell 133:164–176.

  8. Soeno K, Goda H, Ishii T et al (2010) Auxin biosynthesis inhibitors, identified by a genomics-based approach, provide insights into auxin biosynthesis. Plant Cell Physiol 51:524–536.

  9. Mashiguchi K, Tanaka K, Sakai T et al (2011) The main auxin biosynthesis pathway in Arabidopsis. Proc Natl Acad Sci U S A 108:18512–18517.

  10. He W, Brumos J, Li H et al (2011) A small-molecule screen identifies L-kynurenine as a competitive inhibitor of TAA1/TAR activity in ethylene-directed auxin biosynthesis and root growth in Arabidopsis. Plant Cell 23:3944–3960.

  11. Narukawa-Nara M, Nakamura A, Kikuzato K et al (2016) Aminooxy-naphthylpropionic acid and its derivatives are inhibitors of auxin biosynthesis targeting l-tryptophan aminotransferase: structure–activity relationships. Plant J 87:245–257.

  12. Nishimura T, Hayashi K, Suzuki H et al (2014) Yucasin is a potent inhibitor of YUCCA, a key enzyme in auxin biosynthesis. Plant J 77:352–366.

  13. Tsugafune S, Mashiguchi K, Fukui K et al (2017) Yucasin DF, a potent and persistent inhibitor of auxin biosynthesis in plants. Sci Rep 7:13992.

  14. Kakei Y, Yamazaki C, Suzuki M et al (2015) Small-molecule auxin inhibitors that target YUCCA are powerful tools for studying auxin function. Plant J 84:827–837.

  15. Doyle SM, Robert S (2014) Using a reverse genetics approach to investigate small-molecule activity. Methods Mol Biol 1056:51–62.

  16. Bower PJ, Brown HM, Purves WK (1978) Cucumber seedling indoleacetaldehyde oxidase. Plant Physiol 61:107–110.

  17. Sugawara S, Hishiyama S, Jikumaru Y et al (2009) Biochemical analyses of indole-3-acetaldoxime-dependent auxin biosynthesis in Arabidopsis. Proc Natl Acad Sci U S A 106:5430–5435.

  18. Takase S, Uchida I, Tanaka H, Aoki H (1986) Synthesis of debromo-8,8a-dihydroflustramine C1, a model synthesis toward amauromine. Tetrahedron 42:5879–5886.

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Correspondence to Yukihisa Shimada .

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Soeno, K., Sato, A., Shimada, Y. (2021). Investigation of Auxin Biosynthesis and Action Using Auxin Biosynthesis Inhibitors. In: Hicks, G.R., Zhang, C. (eds) Plant Chemical Genomics. Methods in Molecular Biology, vol 2213. Humana, New York, NY.

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