5-azaC对‘巨峰’葡萄果实发育阶段mRNA可变剪接的影响

Analysis of Alternative Splicing at Different Developmental Stages of Kyoho Grapes with 5-azaC Treatment

DNA甲基化和mRNA可变剪接在果实发育过程中均发挥重要作用,但两者间的联系和互作关系尚不清楚。基于5-氮杂胞苷(5-azaC,DNA甲基转移酶抑制剂)处理可以使‘巨峰’葡萄成熟期提前,对5-azaC处理后的‘巨峰’RNA-seq测序,分析果实不同发育阶段发生的可变剪接事件。结果表明,3′端可变剪切位点(A3)类型在处理和对照中均最多,外显子互斥(MX)类型最少。可变剪接在不同发育阶段存在特异调节,发育前期可变剪接的调控更为频繁;鉴定到9683个发育过程中保守的可变剪接事件。在处理与对照间有671个可变剪接事件存在明显的剪接变化,功能注释显示其中有14个基因与甲基化修饰相关,表明可变剪接与甲基化修饰协同调控葡萄果实成熟过程。

Both DNA methylation and alternative splicing play important roles in fruit development,but the links and the interactions between them during fruit development are unclear.In this study,alternative splicing events occurring at different developmental stages of 5-azacytidine(5-azaC,DNA methyltransferase inhibitors)treated and control berries of Kyoho from 25 to 65 days after flowering were analyzed based on RNA-seq data.The results showed that alternative 3′splice site(A3)was the most frequent type and mutually exclusive exons(MX)was the least type revealed in both the treatment and control groups.There was stage-specific regulation of alternative splicing during grape berry development.The events of alternative splicing were more abundant at the early stages of berry development,and 9683 was identified as the conserved events during berry development.There were 671 alternative splicing events which showed significant deviation between the treatments and controls,and functional annotation demonstrated that 14 of these genes were associated with methylation modifications.The significant differences in the extent of alternative splicing of these methylation-related genes in the treated and control groups suggest that alternative splicing and methylation modification synergistically regulate the grape berry ripening process.