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水稻中一个显性矮秆基因d-h的分离与鉴定

Isolation and characterization of a dominant dwarf gene, d-h, in rice.

作者信息

Piao Rihua, Chu Sang-Ho, Jiang Wenzhu, Yu Yoye, Jin Yongmei, Woo Mi-Ok, Lee Joohyun, Kim Sunghan, Koh Hee-Jong

机构信息

Department of Plant Science, Research Institute of Agriculture and Life Sciences and Plant Genomics and Breeding Institute, College of Agriculture and Life Sciences, Seoul National University, Seoul, Korea.

College of Plant Science, Jilin University, Changchun, China.

出版信息

PLoS One. 2014 Feb 3;9(2):e86210. doi: 10.1371/journal.pone.0086210. eCollection 2014.

Abstract

Plant height is an important agronomic trait that affects grain yield. Previously, we reported a novel semi-dominant dwarf mutant, HD1, derived from chemical mutagenesis using N-methyl-N-nitrosourea (MNU) on a japonica rice cultivar, Hwacheong. In this study, we cloned the gene responsible for the dwarf mutant using a map-based approach. Fine mapping revealed that the mutant gene was located on the short arm of chromosome 1 in a 48 kb region. Sequencing of the candidate genes and rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR) analysis identified the gene, d-h, which encodes a protein of unknown function but whose sequence is conserved in other cereal crops. Real-time (RT)-PCR analysis and promoter activity assays showed that the d-h gene was primarily expressed in the nodes and the panicle. In the HD1 plant, the d-h gene was found to carry a 63-bp deletion in the ORF region that was subsequently confirmed by transgenic experiments to be directly responsible for the gain-of-function phenotype observed in the mutant. Since the mutant plants exhibit a defect in GA response, but not in the GA synthetic pathway, it appears that the d-h gene may be involved in a GA signaling pathway.

摘要

株高是影响粮食产量的重要农艺性状。此前,我们报道了一个新的半显性矮化突变体HD1,它是通过对粳稻品种华成用N-甲基-N-亚硝基脲(MNU)进行化学诱变获得的。在本研究中,我们采用图位克隆法克隆了导致该矮化突变体的基因。精细定位表明,突变基因位于第1染色体短臂上一个48 kb的区域。对候选基因进行测序以及通过cDNA末端快速扩增-聚合酶链反应(RACE-PCR)分析鉴定出了d-h基因,该基因编码一种功能未知但在其他谷类作物中序列保守的蛋白质。实时(RT)-PCR分析和启动子活性检测表明,d-h基因主要在节和穗中表达。在HD1植株中,发现d-h基因的开放阅读框(ORF)区域有一个63 bp的缺失,随后通过转基因实验证实该缺失直接导致了突变体中观察到的功能获得型表型。由于突变植株在赤霉素(GA)反应方面存在缺陷,但GA合成途径没有缺陷,因此d-h基因似乎可能参与GA信号传导途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d63f/3911911/d717dd02fe85/pone.0086210.g001.jpg

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