【摘 要】
：
DNA 甲基化作为真核生物中的一种重要表观遗传机制,在不同物种之间的分布模式和作用差异显著.在真菌中,DNA 甲基化在调控多种生物过程中起着重要作用,罗伯茨绿僵菌是研究真菌侵染昆虫机制的一种模式系统,同时也是化学农药杀虫剂的替代品,但是目前对其DNA 甲基化分子机制研究甚少.本论文在罗伯茨绿僵菌中鉴定出两个DNA甲基转移酶(MrRID和MrDIM-2),生物信息学分析发现MrRID和MrDIM-2
【机 构】
：
安徽农业大学,安徽省微生物防治重点实验室,安徽合肥230036 安徽农业大学,安徽省微生物防治重点
【出 处】
：
第六届全国农业微生物研究及产业化研讨会、第十五届全国杀虫微生物学术研讨会、第十一届全国虫生真菌学术研讨会
论文部分内容阅读
DNA 甲基化作为真核生物中的一种重要表观遗传机制,在不同物种之间的分布模式和作用差异显著.在真菌中,DNA 甲基化在调控多种生物过程中起着重要作用,罗伯茨绿僵菌是研究真菌侵染昆虫机制的一种模式系统,同时也是化学农药杀虫剂的替代品,但是目前对其DNA 甲基化分子机制研究甚少.本论文在罗伯茨绿僵菌中鉴定出两个DNA甲基转移酶(MrRID和MrDIM-2),生物信息学分析发现MrRID和MrDIM-2 分别与蛹虫草的CmDMTA和CmDIM-2及脉孢菌中的RID 和DIM-2 同源关系较近.通过根癌农杆菌介导的方法分别不同基因突变菌株(ΔRID、ΔDIM-2 和ΔRID/ΔDIM-2).不同DNA 甲基转移酶突变株的基因组DNA 甲基化结果显示,ΔRID、ΔDIM-2 和ΔRID/ΔDIM-2 菌株中mC位点数量分别占野生型菌株的～71％,～10％和～8％.深入分析发现,MrRID在DNA 甲基化过程中起着识别DNA 甲基化位点的功能,而且一些DNA 位点的甲基化需要MrRID 和MrDIM-2 共同参与.通过检测不同DNA 甲基转移酶突变株的生物性状发现,ΔMrDIM-2 和ΔRID/ΔDIM-2 的生长速度和产孢能力明显弱于野生型和ΔMrRID,热激或紫外照射后突变株(尤其ΔMrDIM-2 和ΔRID/ΔDIM-2)的孢子萌发率相对于野生型明显下降.但是所有突变株对化学物质耐受力与野生型菌株相比无明显差异.毒力测定实验显示,相对于ΔMrRID(LT50 = 4.6 ± 0.8)和野生型(LT50 = 4.4 ± 0.5)菌株,ΔMrDIM-2(LT50 = 6.5 ± 0.9)和ΔRID/ΔDIM-2(LT50 = 7.3 ± 1.2)的毒力明显下降,而且虫子尸体表面产孢能力也明显减弱.
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