木霉菌对有机磷农药的耐受特性是其作为环境修复微生物降解土壤和水体残留农药污染，以及作为生防真菌与有机磷农药协调使用控制作物病虫害的重要基础。因此，利用RNA-Seq这一"组学"方法，深入研究木霉菌耐受有机磷农药敌敌畏的机理具有重要意义。本研究拟采用RNA-Seq技术，从全基因组水平解析深绿木霉（Trichoderma atroviride）在有机磷农药敌敌畏胁迫下的转录谱。在此基础上克隆深绿木霉耐受敌敌畏胁迫的关键基因，并运用基因功能分析、生化检测、超显微等技术，分析深绿木霉耐受敌敌畏相关基因对木霉菌丝生长特性、超微结构、电解质外渗变化、以及对敌敌畏耐受性和降解率等影响；通过酵母双杂交技术鉴定这些与敌敌畏耐受性相关的多个基因所对应蛋白之间的互作关系。从而为揭示深绿木霉耐受有机磷农药的分子机理奠定重要基础，为最终实现对深绿木霉的基因改良，研制高效木霉菌生防菌剂和环境修复剂提供理论依据。

Trichoderma spp. tolerance to organophosphate pesticides is an important basis for degradation of pesticides in contaminated soils and water as an environmental remediation microbial, as well as for controlling crop pests and diseases as fungal biocontrol agents which allows coordinated use with organophosphate pesticides. Therefore, it is of significance to study the mechanism of Trichoderma atroviride tolerance to organophosphate dichlorvos by the new omics tools such as RNA-Seq. In this study, we will analyze the transcriptome profiles of Trichoderma atroviride stressed by dichlorvos using RNA-Seq. The key genes of Trichoderma atroviride tolerance to dichlorvos will be cloned. The functions of the tolerance related genes will be analyzed using gene knockout, biochemical assays and ultramicroscopic techniques. And the interaction between the proteins encoded by tolerance genes will be identified by yeast two-hybrid system. Ultimately, the study lays an important foundation for revealing the molecular mechanism of Trichoderma atroviride tolerance to organophosphate dichlorvos. Also, it provides a theoretical basis for the ultimate realization of genetically modified Trichoderma atroviride, the development of efficient Trichoderma biocontrol agents and environmental remediation agents.

木霉菌对有机磷农药的耐受特性是其作为环境修复微生物降解土壤和水体残留农药污染，以及作为生防真菌与有机磷农药协调使用控制作物病虫害的重要基础。因此，利用RNA-Seq这一“组学”方法，深入研究木霉菌耐受有机磷农药敌敌畏的机理具有重要意义。本研究采用RNA-Seq技术，从全基因组水平解析木霉在有机磷农药敌敌畏胁迫下的转录谱。首先提取了12个木霉样品的RNA，构建测序文库，质检合格后进行测序。在此基础上对测序质量进行评估，并分析基因覆盖度，对reads在木霉基因组及基因上的匹配状况进行了统计，对差异表达基因进行了聚类分析。结果发现木霉在有机磷农药敌敌畏胁迫下，有大量基因均发生显著性上调和下调表达的基因，也发现了些新的转录本和可变剪接模式，表明敌敌畏对木霉全基因组表达模式有明显影响。进一步，对敌敌畏胁迫下木霉的显著性差异表达基因进行更深入的基因功能分析，以更加明确其在木霉耐受敌敌畏中的重要作用；对可变剪接的分析结果进行充分挖掘，深入研究可变剪接与木霉生理代谢、耐受性等相关性。