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分析粳稻品种 Kitaake 开花早期的机制和 T-DNA 标签线的产生。

Analysis of the early-flowering mechanisms and generation of T-DNA tagging lines in Kitaake, a model rice cultivar.

机构信息

Crop Biotech Institute, Kyung Hee University, Yongin 446-701, Korea.

出版信息

J Exp Bot. 2013 Nov;64(14):4169-82. doi: 10.1093/jxb/ert226. Epub 2013 Aug 21.

Abstract

As an extremely early flowering cultivar, rice cultivar Kitaake is a suitable model system for molecular studies. Expression analyses revealed that transcript levels of the flowering repressor Ghd7 were decreased while those of its downstream genes, Ehd1, Hd3a, and RFT1, were increased. Sequencing the known flowering-regulator genes revealed mutations in Ghd7 and OsPRR37 that cause early translation termination and amino acid substitutions, respectively. Genetic analysis of F2 progeny from a cross between cv. Kitaake and cv. Dongjin indicated that those mutations additively contribute to the early-flowering phenotype in cv. Kitaake. Because the short life cycle facilitates genetics research, this study generated 10 000 T-DNA tagging lines and deduced 6758 flanking sequence tags (FSTs), in which 3122 were genic and 3636 were intergenic. Among the genic lines, 367 (11.8%) were inserted into new genes that were not previously tagged. Because the lines were generated by T-DNA that contained the promoterless GUS reporter gene, which had an intron with triple splicing donors/acceptors in the right border region, a high efficiency of GUS expression was shown in various organs. Sequencing of the GUS-positive lines demonstrated that the third splicing donor and the first splicing acceptor of the vector were extensively used. The FST data have now been released into the public domain for seed distribution and facilitation of rice research.

摘要

作为一个极早开花的品种,水稻品种 Kitaake 是分子研究的合适模式系统。表达分析显示,开花抑制因子 Ghd7 的转录水平降低,而其下游基因 Ehd1、Hd3a 和 RFT1 的转录水平升高。对已知的开花调控基因进行测序发现,Ghd7 和 OsPRR37 发生了突变,分别导致早期翻译终止和氨基酸取代。Kitaake 和 Dongjin 品种杂交的 F2 后代的遗传分析表明,这些突变在 Kitaake 品种中累加导致了早花表型。由于短的生命周期有利于遗传学研究,这项研究生成了 10000 个 T-DNA 标签系,并推断出 6758 个侧翼序列标签(FST),其中 3122 个是基因内的,3636 个是基因间的。在基因系中,有 367 个(11.8%)插入到以前未标记的新基因中。由于这些系是由含有无启动子的 GUS 报告基因的 T-DNA 生成的,该基因在右边界区域有一个带有三重剪接受体的内含子,因此在各种器官中显示出高效的 GUS 表达。对 GUS 阳性系的测序表明,载体的第三个剪接受体和第一个剪接受体被广泛使用。FST 数据现已公开发布,用于种子分发和促进水稻研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5c13/3808308/e6fd046ba258/exbotj_ert226_f0001.jpg

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