Small‐molecule auxin inhibitors that target <scp>YUCCA</scp> are powerful tools for studying auxin function

Kakei, Yusuke; Yamazaki, Chiaki; Suzuki, Masashi; Nakamura, Ayako; Satô, Akiko; Ishida, Yasuhiro; Kikuchi, Rie; Higashi, Shouichi; Kokudo, Yumiko; Ishii, Takahiro; Soeno, Kazuo; Shimada, Yukihisa

doi:10.1111/tpj.13032

Cited by 79 publications

(52 citation statements)

References 52 publications

(71 reference statements)

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“…IAA was extracted from Arabidopsis seedlings and quantitative analysis of its endogenous level was performed with liquid chromatography tandem mass spectrometry (LC-MS/MS) with [ 2 H 5 ]-IAA as an internal standard, as described by Kakei et al 39) Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis.…”

Section: Analysis Of Endogenous Iaa Levelsmentioning

confidence: 99%

Auxin signaling through SCFTIR1/AFBs mediates feedback regulation of IAA biosynthesis

Takato

Kakei

Mitsui

et al. 2017

Bioscience, Biotechnology, and Biochemistry

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We previously reported that exogenous application of auxin to Arabidopsis seedlings resulted in downregulation of indole-3-acetic acid (IAA) biosynthesis genes in a feedback manner. In this study, we investigated the involvement of the SCF-mediated signaling pathway in feedback regulation of the indole-3-pyruvic acid-mediated auxin biosynthesis pathway in Arabidopsis. Application of PEO-IAA, an inhibitor of the IAA signal transduction pathway, to wild-type seedlings resulted in increased endogenous IAA levels in roots. Endogenous IAA levels in the auxin-signaling mutants axr2-1, axr3-3, and tir1-1afb1-1afb2-1afb3-1 also increased. Furthermore, YUCCA (YUC) gene expression was repressed in response to auxin treatment, and expression of YUC7 and YUC8 increased in response to PEO-IAA treatment. YUC genes were also induced in auxin-signaling mutants but repressed in TIR1-overexpression lines. These observations suggest that the endogenous IAA levels are regulated by auxin biosynthesis in a feedback manner, and the Aux/IAA and SCF-mediated auxin-signaling pathway regulates the expression of YUC genes.

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Section: Analysis Of Endogenous Iaa Levelsmentioning

confidence: 99%

Auxin signaling through SCFTIR1/AFBs mediates feedback regulation of IAA biosynthesis

Takato

Kakei

Mitsui

et al. 2017

Bioscience, Biotechnology, and Biochemistry

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“…The PpYUC11 enzyme assay was conducted as described previously (Kakei et al ., ) with minor modifications. The activity was measured in a reaction mixture containing E. coli lysates (50 μg soluble protein), 3 μ m IPyA, 30 μ m FAD and 1 m m NADPH in Tris−HCl buffer (50 m m , pH 8.0).…”

Section: Methodsmentioning

confidence: 99%

“…Endogenous IAA, and its biosynthetic intermediates and metabolites were measured according to methods described previously (Kakei et al, 2015;Narukawa-Nara et al, 2016) with modifications. Method S1 describes the analysis of IAA-related compounds in detail.…”

Section: Measurement Of Iaa Biosynthesis Intermediates and Metabolitementioning

confidence: 99%

Insertion of a transposon‐like sequence in the 5′‐flanking region of the YUCCA gene causes the stony hard phenotype

et al. 2018

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Melting-flesh peaches produce large amounts of ethylene, resulting in rapid fruit softening at the late-ripening stage. In contrast, stony hard peaches do not soften and produce little ethylene. The indole-3-acetic acid (IAA) level in stony hard peaches is low at the late-ripening stage, resulting in low ethylene production and inhibition of fruit softening. To elucidate the mechanism of low IAA concentration in stony hard peaches, endogenous levels of IAA and IAA intermediates or metabolites were analysed by ultra-performance liquid chromatography-tandem mass spectrometry. Although the IAA level was low, the indole-3-pyruvic acid (IPyA) level was high in stony hard peaches at the ripening stage. These results indicate that YUCCA activity is reduced in ripening stony hard peaches. The expression of one of the YUCCA isogenes in peach, PpYUC11, was suppressed in ripening stony hard peaches. Furthermore, an insertion of a transposon-like sequence was found upstream of the PpYUC11 gene in the 5'-flanking region. Analyses of the segregation ratio of the stony hard phenotype and genotype in F1 progenies indicated that the transposon-inserted allele of PpYUC11, hd-t, correlated with the stony hard phenotype. On the basis of the above findings, we propose that the IPyA pathway (YUCCA pathway) is the main auxin biosynthetic pathway in ripening peaches of 'Akatsuki' and 'Manami' cultivars. Because IAA is not supplied from storage forms, IAAde novo synthesis via the IPyA pathway (YUCCA pathway) in mesocarp tissues is responsible for auxin generation to support fruit softening, and its disruption can lead to the stony hard phenotype.

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“…In land plants, 5-(4-chlorophenyl)-4H-1,2,4-triazole-3-thiol (yucasin), 4-phenoxyphenylboronic acid (PPBo), and 4-biphenylboronic acid (BBo) are potent inhibitors of YUCCA enzymes and inhibit growth of Arabidopsis (Arabidopsis thaliana; Nishimura et al, 2014;Kakei et al, 2015). At 100 mM, each of yucasin, PPBo, and BBo inhibited K. nitens growth ( Fig.…”

Section: Exogenous Auxin and Auxin Transport Inhibitor Inhibit Growthmentioning

confidence: 99%

Primitive Auxin Response without TIR1 and Aux/IAA in the Charophyte Alga Klebsormidium nitens

et al. 2017

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The phytohormone auxin regulates many aspects of growth and development in land plants, but the origin and evolution of auxin signaling and response mechanisms remain largely unknown. Indeed, it remains to be investigated whether auxin-related pathways diverged before the emergence of land plants. To address this knowledge deficit, we analyzed auxin responses in the charophyte alga Klebsormidium nitens NIES-2285, whose ancestor diverged from a green algal ancestor during the evolution of land plants. This strain is the same as Klebsormidium flaccidum NIES-2285, for which the draft genome was sequenced in 2014, and was taxonomically reclassified as K. nitens. This genome sequence revealed genes involved in auxin responses. Furthermore, the auxin indole-3-acetic acid (IAA) was detected in cultures of K. nitens, but K. nitens lacks the central regulators of the canonical auxin-signaling pathway found in land plants. Exogenous IAA inhibited cell division and cell elongation in K. nitens. Inhibitors of auxin biosynthesis and of polar auxin transport also inhibited cell division and elongation. Moreover, exogenous IAA rapidly induced expression of a LATERAL ORGAN BOUNDARIES-DOMAIN transcription factor. These results suggest that K. nitens has acquired the part of the auxin system that regulates transcription and cell growth without the requirement for the central players that govern auxin signaling in land plants.

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Small‐molecule auxin inhibitors that target YUCCA are powerful tools for studying auxin function

Cited by 79 publications

References 52 publications

Auxin signaling through SCFTIR1/AFBs mediates feedback regulation of IAA biosynthesis

Auxin signaling through SCFTIR1/AFBs mediates feedback regulation of IAA biosynthesis

Insertion of a transposon‐like sequence in the 5′‐flanking region of the YUCCA gene causes the stony hard phenotype

Primitive Auxin Response without TIR1 and Aux/IAA in the Charophyte Alga Klebsormidium nitens

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