重寄生真菌盾壳霉pH信号通路中Pal相关基因的功能鉴定

发布时间：2019-01-30 10:01

【摘要】：【目的】研究pH信号通路(Pal)在重寄生真菌盾壳霉与寄主核盘菌互作过程中的作用。【方法】从盾壳霉全基因组信息中分析获得了6个Pal相关基因CmpalA、CmpalB、CmpalC、CmpalF、CmpalH和CmpalI的全编码序列和氨基酸序列,通过PEG介导的原生质转化技术获得了CmpalA、CmpalB、CmpalC、CmpalF和CmpalH等5个基因的敲除突变体,分析这些敲除突变体与野生型在菌落培养性状、重寄生能力、降解草酸能力、产生抗真菌物质能力等方面的差异。【结果】与野生型相比,在pH 6 8的条件下,5个Pal相关基因敲除突变体的菌丝生长受到显著抑制,这说明缺失Pal相关基因使盾壳霉对高pH值环境更加敏感。菌核重寄生试验发现5个Pal相关基因敲除突变体的重寄生能力均显著低于野生型。qRT-PCR试验结果表明,敲除Pal相关基因之后导致重寄生相关酶基因Cmch1、Cmg1和Cmsp1的表达量显著降低,而且pH信号通路下游的CmpacC基因的表达量也显著降低。Pal相关基因敲除突变体在pH 6条件下对草酸盐的降解能力显著高于野生型,同时这5个突变体在pH 8条件下产生抗真菌物质能力也显著高于野生型。【结论】pH信号通路相关基因的缺失影响盾壳霉对环境pH的响应。pH信号通路在盾壳霉与核盘菌互作中发挥重要作用,不仅影响盾壳霉的重寄生作用,而且还影响盾壳霉的草酸降解作用和抗真菌作用。
[Abstract]:[objective] to study the role of pH signaling pathway (Pal) in the interaction between the highly parasitic fungi of Trichoderma auricularis and host Sclerotinia sclerotiorum. [methods] six Pal related genes, CmpalA,CmpalB,CmpalC,CmpalF, were obtained from the genomic information of C. auriculatus. The full coding sequence and amino acid sequence of CmpalH and CmpalI were used to obtain CmpalA,CmpalB,CmpalC,CmpalF and CmpalH knockout mutants by PEG mediated protoplast transformation technique. The characters of these knockout mutants and wild type in colony culture were analyzed. [results] compared with the wild type, the hyphal growth of 5 Pal related gene knockout mutants was significantly inhibited under the condition of pH 6 + 8, but the ability to degrade oxalic acid and to produce antifungal substances were significantly different, [results] compared with the wild type, the hyphal growth of 5 Pal related gene knockout mutants was significantly inhibited. This suggests that the deletion of Pal related genes makes C. capsidii more sensitive to high pH values. The results of the sclerotia heavy parasitism test showed that the ability of the five Pal related gene knockout mutants was significantly lower than that of the wild type. The results of qRT-PCR test showed that the knockout of Pal related genes led to the hyperparasitic enzyme gene Cmch1,. The expression of Cmg1 and Cmsp1 was significantly decreased, and the expression of CmpacC gene downstream of pH signaling pathway was also significantly decreased. The ability of Pal related gene knockout mutant to degrade oxalate was significantly higher than that of wild type under pH 6 condition. At the same time, the ability of the five mutants to produce antifungal substances under pH 8 was also significantly higher than that of wild type. [conclusion] the absence of genes associated with pH signaling pathway affects the response of C. auriculatus to environmental pH. PH signaling pathway is found in Trichoderma australis. Play an important role in interaction with Sclerotinia sclerotiorum, It not only affects the hyperparasitism, but also the oxalic acid degradation and the antifungal action.
【作者单位】：华中农业大学植物科学技术学院湖北省作物病害监测与安全控制重点实验室;
【基金】：国家自然科学基金(31272084,31471813,31672073)~~
【分类号】：S476

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