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利用 cDNA-AFLP 技术研究盐胁迫下胡杨杂种(银×黑杨)差异表达基因。

Differentially expressed genes in Populus simonii x Populus nigra in response to NaCl stress using cDNA-AFLP.

机构信息

Key Laboratory of Forest Tree Genetic Improvement and Biotechnology of Ministry of Education, Northeast Forestry University, Harbin 150040, China.

出版信息

Plant Sci. 2011 Jun;180(6):796-801. doi: 10.1016/j.plantsci.2011.02.001. Epub 2011 Feb 16.

Abstract

Salinity is an important environmental factor limiting growth and productivity of plants, and affects almost every aspect of the plant physiology and biochemistry. The objective of this study was to apply cDNA-AFLP and to identify differentially expressed genes in response to NaCl stress vs. no-stress in Populus simonii x Populus nigra in order to develop genetic resources for genetic improvement. Selective amplification with 64 primer combinations allowed the visualization of 4407 transcript-derived fragments (TDFs), and 2027 were differentially expressed. Overall, 107 TDFs were re-sequenced successfully, and 86 unique sequences were identified in 10 functional categories based on their putative functions. A subset of these genes was selected for real-time PCR validation, which confirmed the differential expression patterns in the leaf tissues under NaCl stress vs. no stress. Differential expressed genes will be studied further for association with salt or drought-tolerance in P. simonii x P. nigra. This study suggests that cDNA-AFLP is a useful tool to serve as an initial step for characterizing transcriptional changes induced by NaCl salinity stress in P. simonii x P. nigra and provides resources for further study and application in genetic improvement and breeding. All unique sequences have been deposited in the Genbank as accession numbers GW672587-GW672672 for public use.

摘要

盐度是限制植物生长和生产力的重要环境因素,几乎影响植物生理学和生物化学的各个方面。本研究的目的是应用 cDNA-AFLP 技术,鉴定毛白杨杂种与对照在 NaCl 胁迫下差异表达的基因,为遗传改良开发遗传资源。用 64 对引物组合进行选择性扩增,可观察到 4407 个转录衍生片段(TDFs),其中 2027 个差异表达。总的来说,成功地对 107 个 TDFs 进行了测序,根据它们的推测功能,将它们分为 10 个功能类别,共鉴定了 86 个独特序列。选择其中一部分基因进行实时 PCR 验证,证实了 NaCl 胁迫与非胁迫下叶片组织中的差异表达模式。将进一步研究差异表达基因与毛白杨杂种的耐盐性或耐旱性的关系。本研究表明,cDNA-AFLP 是一种有用的工具,可以作为研究 NaCl 盐胁迫诱导的毛白杨杂种转录变化的初始步骤,并为进一步的研究和遗传改良及育种中的应用提供资源。所有独特序列已作为 GW672587-GW672672 号在 Genbank 中注册,供公众使用。

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