草菇热激蛋白60基因(Vvhsp60)生物信息学分析及低温下的表达

Bioinformatics analysis and relative expression of Vvhsp60 gene in Volvariella volvacea under low temperature stress

摘要:

【背景】生物受到温度胁迫时，热激蛋白被诱导并在短时间内大量产生，可以使受损的蛋白质恢复正常构象，增强生物对逆境胁迫的耐受性。【目的】初步探究草菇热激蛋白60(Vvhsp60)与低温耐受性的关系，为深入开展草菇不耐低温特性的遗传改良奠定理论基础。【方法】对Vvhsp60进行生物信息学分析，以低温敏感型草菇菌株V23及耐低温菌株VH3为实验材料，利用实时荧光定量PCR技术分析低温胁迫及热激诱导后在低温下草菇菌丝体中Vvhsp60基因的表达水平。【结果】草菇Vvhsp60编码蛋白不存在信号肽，不属于分泌蛋白，在线粒体和细胞质内发挥生物学作用，属于双向跨膜蛋白。低温处理显著提高了V23与VH3菌丝体中Vvhsp60基因的表达量，而且VH3中的表达量显著高于V23，推测Vvhsp60基因的表达量高可能有助于增强草菇对低温胁迫的耐受性。经热激处理后两菌株Vvhsp60基因的表达量显著高于各自未热激处理的对照组，表明热激处理可诱导Vvhsp60基因的表达。【结论】Vvhsp60与草菇低温耐受性相关，并且热激可以诱导Vvhsp60基因的表达。

Abstract:

[Background] Facing temperature stress, organisms produce a large amount of heat shock protein in a short time, which can restore the conformations of damaged proteins and enhance the tolerance of organisms to stress. [Objective] To preliminarily explore the relationship between heat shock protein Vvhsp60 and low temperature tolerance of Volvariella volvacea, and thereby lay a theoretical foundation for further genetic improvement of V. volvacea for low-temperature tolerance. [Methods] The bioinformation of Vvhsp60 was analyzed. The expression of Vvhsp60 gene in low temperature-sensitive strain V23 and low temperature-tolerant strain VH3 under low temperature stress and under heat shock-low temperature treatment was analyzed by real-time fluorescent quantitative PCR (RT-qPCR). [Results] With no signal peptide, Vvhsp60 is not a secretory protein. It exerts biological functions in mitochondria and cytoplasm and is a bidirectional transmembrane protein. Low temperature significantly increased the expression of Vvhsp60 gene in mycelia of both V23 and VH3, and the expression in VH3 was significantly higher than that in V23, which suggests that the high expression of Vvhsp60 gene may help enhance the low-temperature tolerance of VH3. After heat shock treatment, the expression of Vvhsp60 gene in the two strains was significantly higher than that in those without heat shock treatment, indicating that heat shock can induce the expression of Vvhsp60 gene. [Conclusion] Vvhsp60 gene is related to the low-temperature tolerance of V. volvacea, and heat shock can induce the expression of Vvhsp60 gene.