YLR358C对酿酒酵母细胞壁完整性的调控

YLR358C Regulates Cell Wall Integrity of Saccharomyces cerevisiae

目的观察酿酒酵母YLR358C缺失对细胞壁完整性的调控作用。方法通过点板实验观察YLR358CΔ酵母的生长情况;Calcofluor White(CFW)染色及透射电镜分析YLR358C对酵母细胞壁的影响;转录组分析及qRT-PCR检测YLR358C调控的信号通路。结果在含有不同浓度的细胞壁干扰剂CFW、刚果红(CR)和十二烷基硫酸钠(SDS)的培养基上,YLR358C敲除酿酒酵母的生长受到明显抑制。CFW染色显示YLR358CΔ酵母细胞壁几丁质下调,透射电镜也观察到细胞壁厚度变薄。转录组测序和分析表明,YLR358C基因可能参与CWI信号通路的调控。qRT-PCR检测发现YLR358C敲除后WSC3、SWI4和HSP12有差异表达(P<0.05)。结论YLR358C可能调控酵母细胞壁的完整性,在发酵工程中具有一定的应用潜力。

Objective To observe the effect of Saccharomyces cerevisiae(S.cerevisiae)YLR358C deletion on cell wall integrity.Methods The growth of YLR358CΔyeast was observed by spot assay;the effect of YLR358C on yeast cell wall was analyzed by Calcofluor White(CFW)staining and transmission electron microscopy(TEM);the signal pathway regulated by YLR358C was detected by transcriptome analysis and qRT-PCR.Results The growth of S.cerevisiae knockout with YLR358C was significantly inhibited on medium containing CFW,congored(CR)and sodium dodecyl sulfate(SDS)at different concentrations.The cell wall chitin was downregulated by CFW staining and the cell wall thickness was reduced by TEM.Transcriptome sequencing and analysis indicated that YLR358C gene may be involved in the regulation of CWI signaling pathway.qRT-PCR showed that WSC3,SWI4 and HSP12 were differentially expressed after YLR358C knockout(P<0.05).Conclusion YLR358C may regulate the integrity of yeast cell wall,and has certain application potential in fermentation.