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Deep sequencing of Suppression Subtractive Hybridisation drought and recovery libraries of the non-model crop Trifolium repens L.

机译:非模式作物白三叶的抑制消减杂交干旱和恢复文库的深度测序。

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摘要

Background White clover is a short-lived perennial whose persistence is greatly affected by abiotic stresses, particularly drought. An understanding of its molecular response to water deficit and recovery following re-hydration is of paramount importance to identify targets for the breeding of tolerant varieties. Results We created a white clover reference transcriptome of 16,193 contigs by de novo and reference-based assembly of Illumina reads from four SSH libraries (forward and reverse libraries for each treatment) constructed from leaf material of white clover at the onset of drought and recovery. Reads from individual libraries were then mapped to the reference transcriptome and processed comparing expression level data to generate four robust sets of transcripts induced and repressed in the leaves subjected to and recovering from water stress. The differentially expressed transcripts were functionally annotated and mapped to biological processes and pathways. Conclusions The work presented in this paper provides a simple pipeline for the processing of data generated by deep sequencing of SSH libraries and the first large scale molecular analysis of the white clover response to drought stress and recovery.
机译:背景技术白三叶草是短命的多年生植物,其持久性受到非生物胁迫(尤其是干旱)的极大影响。了解其对补水后水分缺乏和恢复的分子响应,对于确定耐性品种的育种目标至关重要。结果我们通过从头开始的白三叶草参考转录组创建了一个16,193重叠群的白三叶草参考转录组,并在干旱和恢复时从白三叶草的叶片材料构建的四个SSH库(每种处理的正向和反向库)基于Illumina读取的参考组装。然后将来自各个文库的读数映射到参考转录组,并比较表达水平数据进行处理,以生成在受到水分胁迫并从水分胁迫中恢复的叶片中诱导和抑制的四组强大的转录本。对差异表达的转录本进行功能注释,并定位到生物学过程和途径。结论本文提供的工作为处理由SSH文库进行深度测序生成的数据提供了一条简单的流程,并为白三叶草对干旱胁迫和恢复的响应进行了首次大规模分子分析。

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