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Transcriptome characterization and differential expression analysis of cold-responsive genes in young spikes of common wheat

机译:普通小麦幼穗中冷响应基因的转录组表征和差异表达分析

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With the frequent occurrence of climatic anomalies, spring frost has become a significant limiting factor on wheat production, especially during the reproductive growth stage. A high-throughput sequencing technology was applied and a total of 54 million clean reads that corresponded to 7.44 Gb of total nucleotides were generated. These reads were then de novo assembled into 120,715 unigenes with an average length of 627 bp. Functional annotations were then obtained by aligning all unigenes with public protein databases. In total, 9657 potential EST-SSRs were identified, and 6310 primer pairs for 1329 SSRs were obtained. Meanwhile, a comparison of four tag-based digital gene expression libraries, which was built from the control and cold-treated young spikes were performed. Overall, 526 up-regulated and 489 down-regulated genes were identified, and GO and KEGG pathway analyses of those genes were further conducted. Based on these results, a series of candidate genes involved in cold response pathways were identified, and 12 of them were confirmed by qRT-PCR. The combination of RNA-Seq and digital gene expression analysis in this study provides a powerful approach for investigating the transcriptional changes and obtained a large number of unigenes annotated to public databases. (C) 2014 Elsevier B.V. All rights reserved.
机译:随着气候异常的频繁发生,春季霜冻已成为限制小麦产量的重要限制因素,尤其是在生育期。应用了高通量测序技术,并产生了总计5400万个干净的读数,相当于7.44 Gb的总核苷酸。然后将这些读数从头开始组装成120,715个单基因,平均长度为627 bp。然后通过将所有单基因与公共蛋白质数据库比对获得功能注释。总共鉴定出9657个潜在的EST-SSR,并获得了1310个SSR的6310个引物对。同时,比较了四个基于标记的数字基因表达文库,这些文库是由对照和冷处理的幼穗组成的。总体上,鉴定了526个上调基因和489个下调基因,并对这些基因进行了GO和KEGG通路分析。根据这些结果,鉴定了一系列参与冷应答途径的候选基因,并通过qRT-PCR确认了其中的12个。 RNA-Seq和数字基因表达分析的结合在这项研究中提供了一种强有力的方法来研究转录变化,并获得了大量注释到公共数据库的单基因。 (C)2014 Elsevier B.V.保留所有权利。

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