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CYP714B1 和 CYP714B2 编码赤霉素 13-氧化酶,可降低水稻中的赤霉素活性。

CYP714B1 and CYP714B2 encode gibberellin 13-oxidases that reduce gibberellin activity in rice.

机构信息

RIKEN Plant Science Center, Tsurumi, Yokohama, Kanagawa 230-0045, Japan.

出版信息

Proc Natl Acad Sci U S A. 2013 Jan 29;110(5):1947-52. doi: 10.1073/pnas.1215788110. Epub 2013 Jan 14.

Abstract

Bioactive gibberellins (GAs) control many aspects of growth and development in plants. GA(1) has been the most frequently found bioactive GA in various tissues of flowering plants, but the enzymes responsible for GA(1) biosynthesis have not been fully elucidated due to the enzymes catalyzing the 13-hydroxylation step not being identified. Because of the lack of mutants defective in this enzyme, biological significance of GA 13-hydroxylation has been unknown. Here, we report that two cytochrome P450 genes, CYP714B1 and CYP714B2, encode GA 13-oxidase in rice. Transgenic Arabidopsis plants that overexpress CYP714B1 or CYP714B2 show semidwarfism. There was a trend that the levels of 13-OH GAs including GA(1) were increased in these transgenic plants. Functional analysis using yeast or insect cells shows that recombinant CYP714B1 and CYP714B2 proteins can convert GA(12) into GA(53) (13-OH GA(12)) in vitro. Moreover, the levels of 13-OH GAs including GA(1) were decreased, whereas those of 13-H GAs including GA(4) (which is more active than GA(1)) were increased, in the rice cyp714b1 cyp714b2 double mutant. These results indicate that CYP714B1 and CYP714B2 play a predominant role in GA 13-hydroxylation in rice. The double mutant plants appear phenotypically normal until heading, but show elongated uppermost internode at the heading stage. Moreover, CYP714B1 and CYP714B2 expression was up-regulated by exogenous application of bioactive GAs. Our results suggest that GA 13-oxidases play a role in fine-tuning plant growth by decreasing GA bioactivity in rice and that they also participate in GA homeostasis.

摘要

生物活性赤霉素 (GA) 控制着植物生长和发育的许多方面。GA(1) 是各种开花植物组织中最常发现的生物活性 GA,但由于催化 13-羟化步骤的酶尚未鉴定,因此尚未完全阐明负责 GA(1) 生物合成的酶。由于缺乏在该酶中缺失的突变体,GA 13-羟化的生物学意义尚不清楚。在这里,我们报道两个细胞色素 P450 基因 CYP714B1 和 CYP714B2 在水稻中编码 GA 13-氧化酶。过表达 CYP714B1 或 CYP714B2 的转基因拟南芥植物表现出半矮化。这些转基因植物中 13-OH GA(包括 GA(1))的水平呈上升趋势。使用酵母或昆虫细胞进行的功能分析表明,重组 CYP714B1 和 CYP714B2 蛋白可以在体外将 GA(12)转化为 GA(53)(13-OH GA(12))。此外,在水稻 cyp714b1 cyp714b2 双突变体中,包括 GA(1)在内的 13-OH GAs 的水平降低,而包括 GA(4)(比 GA(1)更活跃)在内的 13-H GAs 的水平增加。这些结果表明 CYP714B1 和 CYP714B2 在水稻中主要参与 GA 13-羟化。双突变体植物在抽穗前外观正常,但在抽穗期表现出伸长的最上部节间。此外,外源施用生物活性 GAs 可上调 CYP714B1 和 CYP714B2 的表达。我们的结果表明,GA 13-氧化酶通过降低水稻中 GA 的生物活性在精细调节植物生长中发挥作用,并且它们还参与 GA 动态平衡。

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