Screening and characterization of an inhibitory chemical specific to Arabidopsis gibberellin 2-oxidases

Otani, Masato; Yoon, Jung Min; Park, Seung-Hyun; Asami, Tadao; Nakajima, Miki

doi:10.1016/j.bmcl.2010.05.015

Cited by 7 publications

(10 citation statements)

References 23 publications

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“…2-ODD 19 and 2-ODD 21 showed a 50% decrease in M2H activity in the presence of 5 lM prohexadione-Ca, whereas 2-ODD 11 and 2-ODD 33 showed only slight inhibition of M2H activity. Similar inhibition by prohexadione-Ca has been reported in other 2-ODD enzymes, such as gibberellin 2-oxidase [48].…”

Section: Resultssupporting

confidence: 82%

Molecular cloning of melatonin 2‐hydroxylase responsible for 2‐hydroxymelatonin production in rice (Oryza sativa)

Byeon

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2015

Journal of Pineal Research

103

126

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Although melatonin biosynthetic genes from plants have been cloned, the melatonin catabolism mechanisms remain unclear. To clone the genes responsible for melatonin metabolism, we ectopically expressed 35 full-length cDNAs of rice 2-oxoglutarate-dependent dioxygenase (2-ODD) in Escherichia coli and purified the corresponding recombinant proteins. In vitro 2-ODD assays showed four independent 2-ODD proteins that were able to catalyze melatonin into 2-hydroxymelatonin, exhibiting melatonin 2-hydroxylase (M2H). These M2H proteins had peak activities at pH 8.0 and 30°C. The Km ranged from 121 μm to 371 μm with the Vmax ranging from 1.7 to 18.5 pkat/mg protein, respectively. The M2H enzyme activities were dependent on cofactors such as α-ketoglutarate, ascorbate, and Fe(2+), similar to the 2-ODD enzymes. M2H activity was inhibited by prohexadione-Ca, an inhibitor of 2-ODD, in a dose-dependent manner. M2H activity was high in the roots of rice seedlings, concurrent with high transcription levels of 2-ODD 21, suggesting that 2-ODD 21 was a major gene for M2H activity. Analogous to the high M2H activity in the roots, 2-hydroxymelatonin was found in large quantities in roots treated with melatonin. These results suggest that melatonin was metabolized into 2-hydroxymelatonin by the M2H genes in plants, but the physiological significance of 2-hydroxymelatonin remains to be examined in the future.

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Section: Resultssupporting

confidence: 82%

Molecular cloning of melatonin 2‐hydroxylase responsible for 2‐hydroxymelatonin production in rice (Oryza sativa)

Byeon

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2015

Journal of Pineal Research

103

126

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“…After confirmation by sequence analysis, the resultant vectors were used for protein expression. The expression vector of GA2ox2 was previously described (Otani et al, 2010).…”

Section: Recombinant 2oddsmentioning

confidence: 99%

The apple gene responsible for columnar tree shape reduces the abundance of biologically active gibberellin

et al. 2020

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Ectopic expression of the apple 2-oxoglutarate-dependent dioxygenase (DOX, 2ODD) gene, designated MdDOX-Co, is thought to cause the columnar shape of apple trees. However, the mechanism underlying the formation of such a unique tree shape remains unclear. To solve this problem, we demonstrated that Arabidopsis thaliana overexpressing MdDOX-Co contained reduced levels of biologically active gibberellin (GA) compared with wild type. In summary: (i) with biochemical approaches, the gene product MdDOX-Co was shown to metabolize active GA A 4 (GA 4) to GA 58 (12-OH-GA 4) in vitro. MdDOX-Co also metabolized its precursors GA 12 and GA 9 to GA 111 (12-OH-GA 12) and GA 70 (12-OH-GA 9), respectively; (ii) Of the three 12-OH-GAs, GA 58 was still active physiologically, but not GA 70 or GA 111 ; (iii) Arabidopsis MdDOX-Co OE transformants converted exogenously applied deuterium-labeled (d 2)-GA 12 to d 2-GA 111 but not to d 2-GA 58 , whereas transformants converted applied d 2-GA 9 to d 2-GA 58 ; (iv) GA 111 is converted poorly to GA 70 by GA 20-oxidases in vitro when GA 12 is efficiently metabolized to GA 9 ; (v) no GA 58 was detected endogenously in MdDOX-Co OE transformants. Overall, we conclude that 12-hydroxylation of GA 12 by MdDOX-Co prevents the biosynthesis of biologically active GAs in planta, resulting in columnar phenotypes.

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“…Prohexadione-Ca (PHX, 36), a non-specific GA biosynthesis inhibitor, not only inhibits GA biosynthesis in rice at the 3β hydroxylation step of GA but also inhibits GA catabolism at 2β hydroxylation (Figure 2(A) and (B)) [44]. To stabilize bioactive GAs in plants, Otani et al [45] conducted a chemical screening to identify inhibitors of GA2ox, a deactivating enzyme of bioactive GAs. antagonist of GID1 based on a chemical library screening [64].…”

Section: Auxinmentioning

confidence: 99%

Chemical regulators of plant hormones and their applications in basic research and agriculture*

Jiang

Asami

2018

Bioscience, Biotechnology, and Biochemistry

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119

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Plant hormones are small molecules that play versatile roles in regulating plant growth, development, and responses to the environment. Classic methodologies, including genetics, analytic chemistry, biochemistry, and molecular biology, have contributed to the progress in plant hormone studies. In addition, chemical regulators of plant hormone functions have been important in such studies. Today, synthetic chemicals, including plant growth regulators, are used to study and manipulate biological systems, collectively referred to as chemical biology. Here, we summarize the available chemical regulators and their contributions to plant hormone studies. We also pose questions that remain to be addressed in plant hormone studies and that might be solved with the help of chemical regulators.

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Screening and characterization of an inhibitory chemical specific to Arabidopsis gibberellin 2-oxidases

Cited by 7 publications

References 23 publications

Molecular cloning of melatonin 2‐hydroxylase responsible for 2‐hydroxymelatonin production in rice (Oryza sativa)

Molecular cloning of melatonin 2‐hydroxylase responsible for 2‐hydroxymelatonin production in rice (Oryza sativa)

The apple gene responsible for columnar tree shape reduces the abundance of biologically active gibberellin

Chemical regulators of plant hormones and their applications in basic research and agriculture*

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