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小麦抗条锈病近等基因系与感病近等基因系受小麦条锈菌侵染后的基因表达差异比较分析。

Comparative gene expression analysis of susceptible and resistant near-isogenic lines in common wheat infected by Puccinia triticina.

机构信息

Kihara Institute for Biological Research, Yokohama City University, Yokohama 244-0813, Japan.

出版信息

DNA Res. 2010 Aug;17(4):211-22. doi: 10.1093/dnares/dsq009. Epub 2010 Apr 1.

Abstract

Gene expression after leaf rust infection was compared in near-isogenic wheat lines differing in the Lr10 leaf rust resistance gene. RNA from susceptible and resistant plants was used for cDNA library construction. In total, 55 008 ESTs were sequenced from the two libraries, then combined and assembled into 14 268 unigenes for further analysis. Of these ESTs, 89% encoded proteins similar to (E value of < or =10(-5)) characterized or annotated proteins from the NCBI non-redundant database representing diverse molecular functions, cellular localization and biological processes based on gene ontology classification. Further, the unigenes were classified into susceptible and resistant classes based on the EST members assembled from the respective libraries. Several genes from the resistant sample (14-3-3 protein, wali5 protein, actin-depolymerization factor and ADP-ribosylation factor) and the susceptible sample (brown plant hopper resistance protein, caffeic acid O-methyltransferase, pathogenesis-related protein and senescence-associated protein) were selected and their differential expression in the resistant and susceptible samples collected at different time points after leaf rust infection was confirmed by RT-PCR analysis. The molecular pathogenicity of leaf rust in wheat was studied and the EST data generated made a foundation for future studies.

摘要

在 Lr10 叶锈病抗性基因不同的近等基因小麦系中比较了叶锈病感染后的基因表达。使用感病和抗病植物的 RNA 构建 cDNA 文库。总共从两个文库中测序了 55008 个 ESTs,然后将它们组合并组装成 14268 个 unigenes 进行进一步分析。这些 ESTs 中,89%编码的蛋白质与来自 NCBI 非冗余数据库的(E 值 <或 =10(-5)) 相似,代表了基于基因本体分类的不同分子功能、细胞定位和生物学过程的有特征或注释的蛋白质。此外,根据各自文库组装的 EST 成员将 unigenes 分为敏感和抗性两类。从抗性样本中选择了几个基因(14-3-3 蛋白、wali5 蛋白、肌动蛋白解聚因子和 ADP-核糖基化因子)和敏感样本中的基因(褐飞虱抗性蛋白、咖啡酸-O-甲基转移酶、病程相关蛋白和衰老相关蛋白),并通过 RT-PCR 分析证实了它们在叶锈病感染后不同时间点的抗性和敏感样本中的差异表达。研究了小麦叶锈病的分子致病性,并为未来的研究奠定了 EST 数据基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/519d/2920755/c6254677cf9e/dsq00901.jpg

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