草菇冷诱导基因Cor4在低温下表达变化的研究

Study on the expression of cold induced gene Cor4 of Volvariella volvacea at low temperature

将草菇菌株V23和耐低温诱变草菇菌株VH3的菌丝体在4℃低温条件下处理不同时间后,分别提取总RNA,反转录获得cDNA作为待测模板,构建含Cor4基因和GPD内标基因片段的质粒作为标准品,采用荧光定量PCR技术中的双标准曲线法研究了Cor4基因在低温下的表达变化。结果表明:草菇V23菌株的Cor4基因未经低温处理的表达量最大,低温处理后表达量持续下降,在6 h达到最低值,8 h和10 h该基因的表达量略有回升;而VH3菌株Cor4基因低温处理后表达量呈上升趋势,至4 h达最高值,6 h时略有下降,8 h达最低值,10 h又开始上升,8 h和10 h均低于0 h表达量。

Mycelia of both Volvariella volvacea strains V23 and VH3 were used as test materials and treated at 4℃ for 0, 2, 4, 6, 8 and 10 h, and then the total RNAs were extracted respectively from them. The cDNAs obtained through inverse transcription were used as templates, the plasmids containing Cor4 gene and GPD internal standard gene fragments were constructed as standard, and the expression of Cor4 gene at low temperature was investigated by the two standard curve method of fluoroescent quantitative PCR technology. The results showed that the expression of Cor4 gene in the strain V23 reached the maximum at 0 h low temperature treatment, decreased gradually with low temperature treatment time, and reached the minimum at 6 h, and then increased slightly at 8 and 10 h; The expression of Cor4 gene in VH3 increased gradually with low temperature treatment time, and reached the maximum at 4 h, and later decreased slightly at 6 h, and reached the minimum at 8 h, and then increased again at 10 h; The expressions at 8 and 10 h were less than that at 0 h. It was preliminarily concluded from the above results that Cor4 gene expression correlated with a cold-resistant characteristic of V. volvacea, which would help to further study on breeding a new variety of V. volvacea through genetic engineering.