麦长管蚜9个miRNA在不同发育阶段的表达  

Expression profiles of nine miRNAs in different developmental stages of the grain aphid,Sitobion avenae(Hemiptera:Aphididae)

作　　者：张方梅[1,2] 闫艺 程登发[1] 朱勋[1] 张云慧[1] 李祥瑞[1] ZHANG Fang-Mei;YAN Yi;CHENG Deng-Fa;ZHU Xun;ZHANG Yun-Hui;LI Xiang-Rui(State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China;Xinyang Agriculture and Forestry University, Xinyang, Henan 464000, China)

机构地区：[1]中国农业科学院植物保护研究所,植物病虫害生物学国家重点实验室,北京100193 [2]信阳农林学院,河南信阳464000

出　　处：《昆虫学报》2022年第1期44-52,共9页Acta Entomologica Sinica

基　　金：国家自然科学基金面上项目(31772163);财政部和农业农村部:国家现代农业产业技术体系CARS-03;河南省青年科学基金项目(212300410229)。

摘　　要：【目的】探讨麦长管蚜Sitobion avenae 9个微小RNA(microRNA,miRNA)在两翅型间不同发育阶段的表达模式,揭示其在蚜虫翅型分化中发挥作用的关键时期。【方法】RT-PCR克隆麦长管蚜内参基因U6的cDNA全长序列和9个miRNA,并利用qRT-PCR方法检测9个miRNA在有翅和无翅麦长管蚜不同发育阶段(伪胚胎、1-4龄若蚜和成蚜)的表达。【结果】麦长管蚜U6的cDNA全长为92 bp,与其他昆虫的U6 cDNA序列一致性达80%以上。qRT-PCR结果表明,大部分miRNA在伪胚胎期及若蚜期的相对表达量均较低,在成蚜期相对表达量较高。6个miRNA(miR-277,miR-9a,miR-7,Let-7,miR-1和miR-315)在有翅成蚜中的相对表达量高于无翅成蚜中的;miR-277和miR-9a在有翅成蚜期的相对表达量较高;除miR-315外,miR-277,miR-9a,miR-7,Let-7和miR-1在成蚜期的相对表达量在两翅型间均呈显著差异,且miR-9a在伪胚胎期和miR-315在3龄若蚜期相对表达量在两翅型间也差异显著。其他3个miRNA(miR-8,PC-3p-2743_844和PC-5p-113190_15)在无翅成蚜中的相对表达量高于有翅成蚜中的,其中miR-8和PC-3p-2743_844在有翅成蚜和无翅成蚜期相对表达量显著高于在其他发育阶段,而PC-5p-113190_15的在无翅蚜3龄若蚜期显著高于在其他发育阶段;miR-8在成蚜期以及PC-3p-2743_844在伪胚胎和成蚜期的相对表达量在两翅型间差异显著,PC-5p-113190_15在3龄若蚜期的相对表达量在两翅型间差异极显著。【结论】麦长管蚜内参基因U6与其他昆虫的U6 cDNA序列具有高度保守性。表达谱分析显示,9个miRNA可能在伪胚胎或成虫阶段在麦长管蚜翅型分化中起调控作用。【Aim】This study aims to clarify the expression profiles of nine microRNAs(miRNAs)in the grain aphid,Sitobion avenae at different developmental stages between winged and wingless morphs,and to define the key stage of the aphid wing dimorphism.【Methods】The full-length cDNA of the reference gene U6 and nine miRNAs of S.avenae were cloned by using RT-PCR.The expression levels of nine miRNAs were detected in winged and wingless S.avenae at different developmental stages(pseudo embryo,1st-4th instar nymph and adult)by using qRT-PCR.【Results】The full-length cDNA of U6 of S.avenae is 92 bp and shows over 80%nucleotide sequence identity with U6 from other reported insects.qRT-PCR results revealed that most miRNAs had low relative expression levels in the pseudo embryo and nymphal stages,and high relative expression levels at the adult stage.The relative expression levels of six miRNAs(miR-277,miR-9a,miR-7,Let-7,miR-1 and miR-315)in winged adults were higher than those in wingless adults.MiR-277 and miR-9a had higher relative expression levels in winged adults.Except miR-315,the relative expression levels of five miRNAs including miR-277,miR-9a,miR-7,Let-7 and miR-1 in S.avenae at the adult stage showed significant difference between the winged and wingless morphs.Moreover,there were significant differences in the relative expression levels of miR-9a in the pseudo embryo and miR-315 in the 3rd instar nymphal stage between the winged and wingless morphs.The other three miRNAs(miR-8,PC-3p-2743_844 and PC-5p-113190_15)showed higher relative expression levels in wingless adults than in winged adults.The relative expression levels of miR-8 and PC-3p-2743_844 in winged and wingless adults were significantly higher than those in the other developmental stages,and that of PC-5p-113190_15 in the 3rd nymphs of wingless aphid was significantly higher than those in the other developmental stages.The relative expression levels of miR-8 in the adult stage and PC-3p-2743_844 in the pseudo embryo and adult stages showed signif

关 键 词：麦长管蚜 MIRNA U6 内参基因 翅型分化 表达谱 

分 类 号：Q966[生物学—昆虫学]