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Cloning and expression profiling of heat shock protein gene PcHsp90 in the citrus red mite,Panonychus citri( Acari: Tetranychidae)
Pages: 411-420
Year: Issue:  4
Journal: Acta Entomologica Sinica

Keyword:  Panonychus citriHsp90cDNA cloningcold stressheat stressdevelopmental stagemRNA expression;
Abstract: 【Aim】To explore the roles of Hsp90 gene in development and response to heat and cold stress in the citrus red mite,Panonychus citri. 【Methods】The full-length c DNA encoding Hsp90 from P. citri was cloned and identified using RT-PCR and RACE technique. Bioinformatics programs were used to analyze the sequence characteristics of the gene. The mRNA expression levels of the gene in this mite at different developmental stages and under cold and heat stress conditions were detected by real-time PCR.【Results】The complete c DNA of the cloned gene Pc Hsp90 with Gen Bank accession no. GQ495086 is2 763 bp in length,and contains an open reading frame of 2 193 bp,which encodes a protein of 730 amino acids,with the molecular weight of 83. 85 k Da and the theoretical isoelectric point of 4. 99. The deduced amino acid sequence contains five highly conserved motifs of Hsp90 family and the cytoplasmic Hsp90 C-terminal region "MEEVD". Phylogenetic analysis showed that Pc Hsp90 clustered firstly with Hsp90 of Tetranychus cinnabarinus, and then with that of Ixodes scapularis, suggesting the close relationship among them. Pc Hsp90 transcripts were all detected at different developmental stages,and the relative mRNA expression level was significantly lower at larval stage than those at nymphal and adult stages( P = 0. 015). No distinct change was found in relative Pc Hsp90 mRNA expression levels in the mites subjected to 0- 10℃ cold stress( P = 0. 492),but a significant up-regulation was observed in the mites subjected to 35- 41℃ heat stress( P = 0. 007); especially when the temperature was up to 41℃,the mRNA level was 6. 75-fold as high as that of the control( 25℃). 【Conclusion】Pc Hsp90 may be involved in development and plays an important role in response to heat stress in P. citri.
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