Functional Characterization of the Steroid Reductase Genes GmDET2a and GmDET2b form Glycine max.

Functional Characterization of the Steroid Reductase Genes GmDET2a and GmDET2b form Glycine max.
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大豆类固醇还原酶基因 GmDET2a 和 GmDET2b 的功能表征

DOI:
10.3390/ijms19030726
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发表时间:
2018-03-03
影响因子:
5.6
通讯作者:
Wang J
Wang J
中科院分区:
生物学2区
文献类型:
--
作者:
Huo W;Li B;Kuang J;He P;Xu Z;Wang J

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油菜素类固醇是植物生长和发育的重要植物激素。在大豆 (Glycine max) 中,已鉴定出 BR 受体,但对编码 BR 生物合成相关酶的基因仍知之甚少。在这里,我们发现大豆基因组编码八种类固醇还原酶(GmDET2a 至 GmDET2h)。系统发育分析将来自苔藓、蕨类植物和高等植物的 105 种类固醇还原酶分为 5 个亚类,表明类固醇还原酶家族经历了纯化选择。 GmDET2a 和 GmDET2b 是拟南芥类固醇 5α-还原酶 AtDET2 的同源物,是 263 个氨基酸的蛋白质。 GmDET2a 和 GmDET2b 的异位表达挽救了 Atdet2-1 突变体在黑暗和光照下的缺陷。与突变体相比,转基因株系GmDET2a和GmDET2b的下胚轴长度和株高在黑暗和光照下均显着增加,并且与Atdet2-1相比,GmDET2aOX-23和GmDET2bOX-16株系中BR生物合成相关基因CPD、DWF4、BR6ox-1和BR6ox-2的转录水平下调。实时定量 PCR 显示,GmDET2a 和 GmDET2b 在所有测试的大豆器官中普遍表达,包括根、叶和下胚轴。此外,表油菜素类固醇负调节 GmDET2a 和 GmDET2b 表达。硫酸盐缺乏下调叶片中的 GmDET2a 以及叶片和根中的 GmDET2b;相比之下,缺磷会上调根和叶中的 GmDET2b。综上所述,我们的结果表明 GmDET2a 和 GmDET2b 具有类固醇还原酶的功能。
Brassinosteroids are important phytohormones for plant growth and development. In soybean (Glycine max), BR receptors have been identified, but the genes encoding BR biosynthesis-related enzymes remain poorly understood. Here, we found that the soybean genome encodes eight steroid reductases (GmDET2a to GmDET2h). Phylogenetic analysis grouped 105 steroid reductases from moss, fern and higher plants into five subgroups and indicated that the steroid reductase family has experienced purifying selection. GmDET2a and GmDET2b, homologs of the Arabidopsis thaliana steroid 5α-reductase AtDET2, are proteins of 263 amino acids. Ectopic expression of GmDET2a and GmDET2b rescued the defects of the Atdet2-1 mutant in both darkness and light. Compared to the mutant, the hypocotyl length and plant height of the transgenic lines GmDET2a and GmDET2b increased significantly, in both darkness and light, and the transcript levels of the BR biosynthesis-related genes CPD, DWF4, BR6ox-1 and BR6ox-2 were downregulated in GmDET2aOX-23 and GmDET2bOX-16 lines compared to that in Atdet2-1. Quantitative real-time PCR revealed that GmDET2a and GmDET2b are ubiquitously expressed in all tested soybean organs, including roots, leaves and hypocotyls. Moreover, epibrassinosteroid negatively regulated GmDET2a and GmDET2b expression. Sulfate deficiency downregulated GmDET2a in leaves and GmDET2b in leaves and roots; by contrast, phosphate deficiency upregulated GmDET2b in roots and leaves. Taken together, our results revealed that GmDET2a and GmDET2b function as steroid reductases.
DOI: 10.1093/bioinformatics/8.3.275
发表时间: 1992-06-01
期刊: COMPUTER APPLICATIONS IN THE BIOSCIENCES
影响因子: --
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JONES, DT;TAYLOR, WR;THORNTON, JM
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期刊: SCIENCE
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DOI: 10.1016/s0092-8674(02)00812-7
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期刊: CELL
影响因子: 64.5
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DOI: 10.1104/pp.103.026195
发表时间: 2003-11-01
期刊: PLANT PHYSIOLOGY
影响因子: 7.4
作者:
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通讯作者: Watanabe, Y
DOI: 10.1007/978-94-007-0189-2_12
发表时间: 2011-01-01
期刊: BRASSINOSTEROIDS: A CLASS OF PLANT HORMONE
影响因子: --
作者:
Hayat, Shamsul;Irfan, Mohammad;Ahmad, Aqil
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