Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (16): 3514-3522.doi: 10.3864/j.issn.0578-1752.2013.16.023

• RESEARCH NOTES • Previous Articles    

Molecular Cloning and Expression Analysis of Heat-Shock-Protein90 (LsHsp90) Gene from Leaf Lettuce (Lactuca sativa L.) Under Heat Shock

 REN  Yue, HAN  Ying-Yan, LI  Ting, HAO  Jing-Hong, FAN  Shuang-Xi   

  1. College of Plant Science and Technology, Beijing University of Agriculture, Beijing 102206
  • Received:2013-02-07 Online:2013-08-15 Published:2013-05-02

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Abstract: 【Objective】 LsHsp90 gene was cloned and used to analyze the expression profiles under heat shock stresses, in order to provide a foundation for investigating heat tolerant mechanisms of molecular regulation in stress tolerance of Lactuca sativa L.【Method】 With homology cloning approaches coupling with RACE techniques, the full-length cDNA of LsHsp90 was cloned from total RNA of the leaves of L. sativa L. Real-time quantitative PCR (qRT-PCR) was used to analyze the expression patterns of this gene in leaves after heat shock unter 37℃ and 42℃.【Result】The full cDNA sequence of LsHsp90 is 2 330 bp，containing a 2 097 bp open reading flame (ORF) and encoding 698 amino acid residues with a predicted molecular mass of 79.8 kD．The LsHsp90 protein contains a predicted ATPase site and a Hsp90 conservative structure domain, which is highly conserved in plants and especially similar to Hsp90 from Ageratina adenophora (EU269070.1) and Arabidopsis thaliana (AY081302.1). Phylogenetic analysis showed that LsHsp90 and Hsp90 from Ageratina adenophora (EU269070.1) were clustered into one group. Real-time PCR analysis showed that, the expression of LsHsp90 in leaves of two varieties was upregulated under 37℃. LsHsp90 was induced down regulation in heat-sensitive variety S106 under 42℃, while the expression of LsHsp90 in heat-tolerant variety Z36 was upregulated.【Conclusion】LsHsp90 gene was firstly isolated and characterized from L. sativa L. This gene has genetic characteristics similar with all species and the induced expression profiling of LsHsp90 showed different patterns under heat stress. The results indicate that the LsHsp90 gene may play an important role in response to heat shock stress.

Key words: Lactuca sativa L. , Hsp90 , gene clone , heat stress , gene expression

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