摘要
以糙皮侧耳(Pleurotus ostreatus)菌株CCMSSC00389为材料,研究高温胁迫(40℃、120 min)下钙离子的响应及其对热激蛋白的调控。在高温胁迫条件下,在培养基中添加胞内钙离子螯合剂、胞外钙离子螯合剂、钙调蛋白抑制剂、质膜钙通道抑制剂、三磷酸肌醇受体拮抗剂,利用钙离子特异性荧光探针检测菌丝体胞内钙离子含量,采用荧光定量PCR检测热激蛋白的基因表达量。结果表明,高温胁迫促进胞内钙离子浓度升高,增加Hsp60、Grp78、Hsp90、Hsp104等4种热激蛋白的基因表达量;培养基中添加钙离子螯合剂或抑制剂会降低胞内钙离子浓度,抑制这些热激蛋白的基因表达。研究结果表明高温胁迫会诱导糙皮侧耳胞内钙离子浓度升高,参与信号转导,从而调控热激蛋白的表达来缓解高温胁迫引起的伤害。
The response of calcium ion(Ca~(2+))and its regulation on heat shock proteins(Hsps)in Pleurotus ostreatus(CCMSSC00389)under heat stress(40 ℃,120 min)were investigated in the study.Under heat stress,an intracellular Ca~(2+) chelator,an extracellular Ca~(2+) chelator,a calmodulin inhibitor,a plasma membrane Ca~(2+) channel inhibitor,and an antagonist of inositol 1,4,5-trisphosphate receptor were supplemented into the medium respectively,then the cytosolic Ca~(2+) was detected by a Ca~(2+) fluorescent probe,and the gene expression levels of Hsps were detected by fluorescence quantitative PCR.It was shown that heat stress induced the increase of cytosolic Ca~(2+) concentration and the gene expression of four kinds of Hsps(Hsp60,Grp78,Hsp90 and Hsp104),while the addition of the Ca~(2+) chelators or inhibitors in the medium decreased the cytosolic Ca~(2+) concentration and inhibited the gene expression of Hsps.The results revealed that cytosolic Ca~(2+) in P.ostreatus participated in signal transduction responsive to heat stress,thereby regulating the gene expression of Hsps to alleviate damages caused by heat stress.
引文
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