马铃薯HSP17.7基因的克隆及其对高温逆境的响应
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福建农林大学园艺学院

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福建省闽江学者科研基金(116-114120019);福建农林大学博士研究生出国合作研究计划资助(324-112110074)


Cloning of HSP17.7 Gene and Its Response to High Temperature Stress in Potato
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Ming Jiang Scholar fund from Fujian Province (116-114120019);Fujian Agriculture and Forestry University of doctoral students going aboard to be joint trained (324-112110074 )

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    摘要:

    热激蛋白和植物对高温的响应密切相关,同时在植物生长发育调控和逆境抵抗等方面具有重要作用。本意见克隆了马铃薯热激蛋白基因StHSP17.7(GenBank登录号为XP_006350804.1),对其全长cDNA序列进行了相关生物信息学分析,利用qRT-PCR技术分析该基因在高温胁迫下的差异表达情况。结果显示:(1)StHSP17.7全长755 bp,包含465 bp开放阅读框,编码154个氨基酸。(2)StHSP17.7分子质量约为17.62 kD,等电点7.91,为亲水性蛋白,C端含有保守序列I和Ⅱ组成的ACD结构域,属于典型的sHSPs家族成员。(3)系统进化分析发现马铃薯小分子热激蛋白归为12个亚家族,其中StHSP17.7与马铃薯HSP17.6蛋白亲缘关系较近,属于小分子热激蛋白C的I类家族成员;基因结构分析显示,在马铃薯48个sHSP基因中,StHSP17.7基因不含内含子,含1个内含子的基因有23个,占47.9%。(4)qRT-PCR分析结果显示,高温快速诱导StHSP17.7基因表达,且基因表达量呈爆发式变化,在24 h达到最高值。StHSP17.7基因明显参与了马铃薯对高温的响应。

    Abstract:

    Plant heat-shock proteins are representative proteins that respond to high temperatures and play an important role in regulating plant growth and development and resisting adversity. In this study, HSP17.7 gene from Solanum tuberosum was cloned by RT-PCR and bioinformatic characters were analyzed. The expression of level of StHSP17.7 gene under high temperature stress was analyzed by qRT-PCR. The results showed that: (1) Full-length sequence of StHSP17.7 gene was 755 bp. It contains an open reading frame of 465 bp and 154 amino acids were encoded. (2) The StHSP17.7 gene encoding protein was a hydrophilic protein with molecular weight of 17.62 kD and the isoelectric point is 7.91. The functional structure domain analysis showed that the C-terminal of the protein contained an ACD domain consisting of conserved sequence Ⅰ and conserved sequence Ⅱ, typically belonging to the sHSPs family member. (3) Phylogenetic analysis showed that sHSPs of Solanum tuberosum was clustered into 12 subfamilies. The StHSP17.7 protein was closely related to the HSP17.6 protein of Solanum tuberosum, and belongs to the family of small heat shock protein C I. Gene structure analysis showed that among 48 StHSPs genes from Solanum tuberosum, StHSP17.7 gene does not contain intron, and other 23 genes contained one introns, accounting for 47.9% of all the genes in the family. (4) qRT-PCR analysis showed that StHSP17.7 gene expression was rapidly induced by high temperatures, and the gene expression level showed an explosive change, reaching the highest value in 24 h. The results suggest that StHSP17.7 gene is very sensitive to heat stress and may play an important role in response to high-temperature stress in potatoes.

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  • 收稿日期:2019-07-20
  • 最后修改日期:2019-10-10
  • 录用日期:2019-10-10
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