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首页> 外文期刊>Plant molecular biology reporter >Construction, Characterization, and Expressed Sequence Tag (EST) Analysis of Normalized cDNA Library of Thermo-Photoperiod-Sensitive Genic Male Sterile (TPGMS) Wheat from Spike Developmental Stages
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Construction, Characterization, and Expressed Sequence Tag (EST) Analysis of Normalized cDNA Library of Thermo-Photoperiod-Sensitive Genic Male Sterile (TPGMS) Wheat from Spike Developmental Stages

机译：穗发育期热光敏基因雄性不育小麦（TPGMS）标准化cDNA文库的构建，表征和表达序列标签（EST）分析

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摘要

Thermo-photoperiod-sensitive genic male sterile (TPGMS) wheat is important in utilization of heterosis. To facilitate the use of such wheat line in agriculture, more knowledge about molecular mechanisms of TPGMS genes is required. In this study, we set up a normalized complementary DNA (cDNA) library based on the strategy of saturation hybridization with genomic DNA using TPGMS wheat line. This normalized cDNA library consists of cDNA from six directionally cloned cDNA libraries constructed with spike and anther tissues from spike developmental stages. From the normalized cDNA library, 3,264 single-pass expressed sequence tag (EST) were obtained. Exclusion of sequences shorter than 100 bp resulted in 3,223 vector-trimmed ESTs with a mean length of 926 bp. Clustering and assembly analysis resulted in 2,175 unique ESTs from 423 contigs and 1,752 singletons. Taking advantage of various tools and database, gene function classification showed that 60% of the ESTs were predicted to have putative gene function. Of the 2,175 unique ESTs, 264 (12%) displayed significant homology (BlastX E values <10e) to genes previously reported to be involved in cold-response related processes. Among these, sequences encoding activities related to primary metabolism, signal transduction, and transcriptional regulation were observed. Finally, in the total EST sequences, 108 potential SSRs were found. The unigene dataset will now be used to fabricate biochips carrying all identified genes for TPGMS wheat functional genomic research.
机译：热光周期敏感的雄性不育小麦（TPGMS）在利用杂种优势中很重要。为了促进这种小麦品系在农业中的使用，需要更多有关TPGMS基因分子机制的知识。在这项研究中，我们基于使用TPGMS小麦品系与基因组DNA饱和杂交的策略，建立了标准化的互补DNA（cDNA）文库。这个标准化的cDNA文库由来自六个定向克隆cDNA文库的cDNA组成，这些文库由来自穗发育阶段的穗和花药组织构成。从标准化的cDNA文库中，获得了3264个单程表达序列标签（EST）。排除短于100 bp的序列会产生3,223个载体修饰的EST，平均长度为926 bp。聚类和组装分析从423个重叠群和1752个单例中产生了2175个唯一的EST。利用各种工具和数据库，基因功能分类显示，预计60％的EST具有推定的基因功能。在2175个独特的EST中，有264个（12％）与先前报道的参与冷应答相关过程的基因显示出显着的同源性（BlastX E值<10e）。在这些序列中，观察到了编码与初级代谢，信号转导和转录调控有关的活性的序列。最后，在全部EST序列中，发现了108个潜在的SSR。现在，单基因数据集将用于制造携带所有已鉴定基因的生物芯片，用于TPGMS小麦功能基因组研究。

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