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水分胁迫下高羊茅转录组的从头组装与特征分析

De Novo Assembly and Characterization of Tall Fescue Transcriptome under Water Stress.

作者信息

Talukder S K, Azhaguvel P, Mukherjee S, Young C A, Tang Y, Krom N, Saha M C

机构信息

The Samuel Roberts Noble Foundation, 2510 Sam Noble Pkwy., Ardmore, OK, 73401.

出版信息

Plant Genome. 2015 Jul;8(2):eplantgenome2014.09.0050. doi: 10.3835/plantgenome2014.09.0050.

Abstract

Water stress is a fundamental problem for tall fescue [Lolium arundinaceum (Schreb.) Darbysh.] cultivation in the south-central United States. Genetic improvement of tall fescue for water-stress tolerance is the key strategy for improving its persistence in the region. Genotypes with contrasting characteristics for relative water content and osmotic potential were identified from a tall fescue population. Transcriptome profiling between water-stress-tolerant (B400) and water-stress-susceptible (W279) genotypes was performed to unravel the genetic regulatory mechanism of water-stress responses in tall fescue. RNA samples from leaf, shoot, root, and inflorescence were pooled and sequenced through Illumina paired-end sequencing. A total of 199,399 contigs were assembled with an average length of 585 bp. Between the two genotypes, 2986 reference transcripts (RTs) were significantly differentially expressed and 1048 of them could be annotated and found to associate with metabolic pathways and enzyme coding genes. In total, 175 differentially expressed RTs were reported for various stress-related functions. Among those, 65 encoded kinase proteins, 40 each encoded transposons, and transporter proteins were previously reported to be involved with abiotic stress responses. A total of 6348 simple sequence repeats and 6658 single-nucleotide polymorphisms were identified in the contig sequences. Primers were developed from the corresponding sequences, which might be used as candidate gene markers in tall fescue. This study will lead to identification of genes or transcription factors related to water-stress tolerance and development of a comprehensive molecular marker system to facilitate marker-assisted breeding in tall fescue.

摘要

水分胁迫是美国中南部地区高羊茅[Lolium arundinaceum (Schreb.) Darbysh.]种植面临的一个基本问题。通过基因改良提高高羊茅的耐水分胁迫能力是增强其在该地区持久性的关键策略。从一个高羊茅群体中鉴定出了相对含水量和渗透势具有不同特征的基因型。对耐水分胁迫(B400)和水分胁迫敏感(W279)基因型进行转录组分析,以揭示高羊茅水分胁迫响应的遗传调控机制。将来自叶片、茎、根和花序的RNA样本混合,并通过Illumina双末端测序进行测序。共组装了199,399个重叠群,平均长度为585 bp。在这两个基因型之间,有2986个参考转录本(RTs)显著差异表达,其中1048个可以注释,并发现与代谢途径和酶编码基因相关。总共报告了175个与各种胁迫相关功能差异表达的RTs。其中,65个编码激酶蛋白,40个分别编码转座子,先前报道转运蛋白与非生物胁迫响应有关。在重叠群序列中总共鉴定出6348个简单序列重复和6658个单核苷酸多态性。从相应序列开发了引物,这些引物可作为高羊茅候选基因标记。本研究将有助于鉴定与耐水分胁迫相关的基因或转录因子,并开发一个综合分子标记系统,以促进高羊茅的标记辅助育种。

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