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利用 cDNA-AFLP 分析鉴定水稻叶片响应热胁迫的差异表达基因。

Identification of differential expression genes in leaves of rice (Oryza sativa L.) in response to heat stress by cDNA-AFLP analysis.

机构信息

College of Life Science, Nantong University, Nantong, Jiangsu 226019, China.

出版信息

Biomed Res Int. 2013;2013:576189. doi: 10.1155/2013/576189. Epub 2013 Feb 17.

Abstract

High temperature impedes the growth and productivity of various crop species. To date, rice (Oryza sativa L.) has not been exploited to understand the molecular basis of its abnormally high level of temperature tolerance. To identify transcripts induced by heat stress, twenty-day-old rice seedlings of different rice cultivars suffering from heat stress were treated at different times, and differential gene expression analyses in leaves were performed by cDNA-AFLP and further verified by real-time RT-PCR. In aggregate, more than three thousand different fragments were indentified, and 49 fragments were selected for the sequence and differential expressed genes were classified functionally into different groups. 6 of 49 fragments were measured by real-time RT-PCR. In addition, the variations of three different polyamine contents in response to heat stress through high-performance liquid chromatography (HPLC) analysis were also performed. The results and their direct and indirect relationships to heat stress tolerance mechanism were discussed.

摘要

高温会阻碍各种作物的生长和生产力。迄今为止,人们尚未利用水稻(Oryza sativa L.)来了解其异常高水平耐热性的分子基础。为了鉴定热胁迫诱导的转录本,对不同水稻品种的 20 天大的水稻幼苗在不同时间进行热胁迫处理,并通过 cDNA-AFLP 进行叶片差异基因表达分析,进一步通过实时 RT-PCR 进行验证。总的来说,鉴定了三千多个不同的片段,选择了 49 个片段进行测序,并根据功能将差异表达基因分类为不同的组。通过实时 RT-PCR 测量了 49 个片段中的 6 个。此外,还通过高效液相色谱(HPLC)分析研究了三种不同多胺含量对热应激的响应变化。讨论了这些结果及其与耐热性机制的直接和间接关系。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/227c/3590577/bfbd3b589853/BMRI2013-576189.001.jpg

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