异辛醇对灰葡萄孢的抑菌作用及转录组分析

Inhibition Effect and Transcriptome Analysis of Isooctyl alcohol on Botrytis cinerea

为探究异辛醇对灰葡萄孢Botrytis cinerea的抑制机理,采用双皿对扣法测定了异辛醇对灰葡萄孢的抑菌效果,利用扫描电镜技术观察了异辛醇处理后灰葡萄孢菌丝的形态变化,测定异辛醇对灰葡萄孢细胞膜胞外电导率和麦角甾醇合成的影响,并基于转录组测序技术(RNA sequencing,RNA-Seq)分析了异辛醇处理对灰葡萄孢的影响。结果表明:异辛醇对灰葡萄孢菌丝的抑菌率为67.92%。经异辛醇处理后,灰葡萄孢菌丝出现大量畸形、不规则变形扭曲,细胞膜通透性增加。异辛醇处理4、5和6 d后,细胞膜麦角甾醇的含量分别降低44.35%、38.82%和42.43%;蛋白质浓度分别降低65.70%、77.93%和58.57%。转录组测序共得到577个差异表达基因(DEGs),其中312个基因表达量上调,265个基因表达量下调。通过同源蛋白簇(COG)、基因本体论(GO)和京都基因与基因组百科全书(KEGG)功能分析,发现参与膜整体组分、麦角甾醇生物合成途径、氨基酸代谢和核糖体生物发生通路中的DEGs均显著下调,表明异辛醇可能通过影响灰葡萄孢的麦角甾醇含量、能量的产生和核糖体结构,从而起到对灰葡萄孢的抑菌作用。

To reveal the inhibitory mechanism of isooctyl alcohol to Botrytis cinerea,the inhibition effect of isooctyl alcohol on B.cinerea was determined using two-sealed-base-plate method.Mycelial morphology treated with isooctyl alcohol was observed by scanning electron microscopy.The influence of isooctyl alcohol on the extracellular conductivity and the ergosterol synthesis were measured.Response of B.cinerea to isooctyl alcohol was analyzed by RNA sequencing(RNA-Seq).The results showed that the inhibition rate of isooctyl alcohol to mycelia was 67.92%.After treatment with isooctyl alcohol,lots of mycelia were deformed and distorted with the increasing of membrane permeability.At 4,5 and 6 days after isooctyl alcohol treatment,the ergosterol contents in cell membrane decreased to 44.35%,38.82%and 42.43%,and the protein concentration decreased to 65.70%,77.93%and 58.57%,respectively.A total of 577 differentially expressed genes(DEGs),including 312 up-regulated genes and 265 down-regulated genes,were obtained by RNA-Seq.Based on functional analysis of clusters of orthologous groups(COG),gene ontology(GO)and kyoto encyclopedia of genes and genomes(KEGG),DEGs involved in membrane integral components,ergosterol biosynthesis pathway,amino acid metabolism and ribosome biogenesis pathway were significantly down-regulated,indicating that isooctyl alcohol might affect ergosterol content,energy production and ribosome structure,and further play inhibit role in the growth of B.cinerea.