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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (25): 20-26.

所属专题： 生物技术

• 林学 园艺 园林 • 上一篇    下一篇

小叶杨热激转录因子基因的克隆及单核苷酸多态性分析

王建义 慈忠玲 王贵 卫尊征   

  • 收稿日期:2010-06-12 修回日期:2010-09-03 出版日期:2012-09-05 发布日期:2012-09-05
  • 基金资助:

    国家林业局948项目

Analysis on Isolation and Single Nucleotide Polymorphisms (SNPs) of Heat Shock Transcription Factor Gene of Populus simonii

  • Received:2010-06-12 Revised:2010-09-03 Online:2012-09-05 Published:2012-09-05

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摘要：

热激转录因子（heat shock transcription factor，HSF）在调节植物对逆境胁迫（特别是热胁迫）应答和相关基因表达方面起重要作用。笔者采用RT-PCR方法，首次从小叶杨基因组DNA中分离出热激转录因子基因，并进行测序和序列分析。结果表明：小叶杨HSF基因总长度为2884 bp，基因内部含有完整的编码阅读框架，即编码区为1398 bp，可编码466个氨基酸残基，所推导的蛋白质序列与拟南芥和葡萄HSF蛋白的同源性分别为63%和78%。组合利用MEGA3.1和DNASP4.0软件对小叶杨16株基因型个体的HSF序列进行比对和分析，共检测到107个SNP位点，多态性频率为1/27 bp，其中常见SNP为42个，罕见SNP为65个。在PsHSFB基因内部发生的单核苷酸变异中，71个属于转换，36个属于颠换。在外显子区域，共检测到52个SNP位点，其中15个为同义突变，37个为错义突变。本研究为小叶杨热激转录因子基因的连锁不平衡作图及其基因辅助小叶杨抗逆性状的分子育种提供重要的理论依据。

关键词: 环洞庭湖区, 环洞庭湖区, 社会主义新农村, 五个文明建设

Abstract:

HSF plays an important role in regulating plant to adversity stress response and related gene expression. In this study, the segments of HSF were isolated from the genomic DNA of Populus simonii by the RT-PCR method. The results showed that: the genomic DNAs were 2884 bp in length with corresponding open reading frames (ORFs) which were capable of encoding the protein of 466 amino acids (aa). The deduced aa sequence of the PsHSF protein shared 63% and 78％ identity with the other recently isolated A. thaliana, and Vitis vinifera HSF1 respectively. The genomic sequences of PsHSF in 16 individuals were aligned, compared and analyzed using the software MEGA3.1 and DNASP4.0. A total of 107 SNPs were detected and the diversity of them was 1/27 bp, in which 42 were common SNPs and 65 were rare SNPs. There were 71 transition and 36 transversion of mutation types. In total, 52 SNPs were detected in the coding regions of PsHSF, of which 15 were nonsense mutation and 37 were missense mutation. Therefore, the results provided the important foundation for association genetics of HSF gene and gene-assisted breeding of new germplasms with drought and high temperature tolerance in P. simonii.