昆虫学报2025，Vol.68Issue(3)：260-270,11.DOI:10.16380/j.kcxb.2025.03.002
意大利蜜蜂piR-ame-1186994的靶基因与功能分析
Target gene and functional analysis of piR-ame-1186994 in Apis mellifera ligustica(Hymenoptera:Apidae)
摘要
Abstract
[Aim]The objective of this study is to elucidate the regulatory function of piR-ame-1186994 of Apis mellifera ligustica,so as to offer a scientific basis for further investigation of the underlying regulatory mechanism of piR-ame-1186994.[Methods]The expression and sequence authenticity of piR-ame-1186994 in the 6-day-old adult worker's midgut,12-day-old adult drone's testis and 7-day-old adult queen's ovary of A.m.ligustica were verified by Stem-loop RT-PCR and Sanger sequencing,respectively.Relevant software was utilized to predict the target mRNAs of piR-ame-1186994 followed by GO and KEGG database annotation.Regulatory networks related to developmental signaling pathways,energy metabolism pathways and cellular and humoral immune pathways were further constructed.Newly emerged adult workers were fed with mimic and mimic-NC(negative control)of piR-ame-1186994,followed by the detection of the relative expression levels of piR-ame-1186994 and its key target genes(YAP1 and PLD2)in the midguts of adult workers using RT-qPCR.[Results]The specific fragment of piR-ame-1186994 was amplified from the 6-day-old adult worker's midgut,12-day-old adult drone's testis and 7-day-old adult queen's ovary of A.m.ligustica.piR-ame-1186994 targeted 1 097 mRNAs,which could be annotated to 30 GO terms involved in metabolic process,binding,cell,etc.,and 182 KEGG pathways including Wnt signaling pathway,endocytosis and oxytocin signaling pathway.Thirty-six and 16 target mRNAs were respectively involved in five developmental-related signaling pathways(mTOR,Wnt,Hippo,AMPK and Notch signaling pathways)and four pathways related to energy metabolism(amino sugar and nucleotide sugar metabolism,fructose and mannose metabolism,glycolysis/gluconeogenesis,and pentose phosphate pathway),respectively.Additionally,29 and eight target mRNAs were engaged in four cellular immune pathways(lysosome,endocytosis,phagosome and ubiquitin-mediated protein degradation)and three humoral immune pathways(PI3K-Akt,MAPK and FoxO signaling pathways),respectively.In the mimic-piR group,the expression level of piR-ame-1186994 was significantly up-regulated in the 1-,3-and 5-day-old workers'midguts and up-regulated extremely significantly in the 2-and 4-day-old workers'midguts,the expression level of the target gene YAP1 was extremely significantly down-regulated in the 1-,3-,4-and 5-day-old workers'midguts and significantly down-regulated in the 2-day-old worker's midgut,and the expression level of target gene PLD2 was significantly down-regulated in the 2-,3-and 5-day-old workers'midguts and down-regulated extremely significantly in the 4-day-old worker's midguts as compared with those in the mimic-NC group.[Conclusion]piR-ame-1186994 exists and expresses in the worker's midgut,drone's testes and queen's ovary of A.m.ligustica.piR-ame-1186994 potentially modulates the development and immunity of worker's midgut through targeting and negatively regulating the expression of YAP1 and PLD2.关键词
意大利蜜蜂/中肠/卵巢/精巢/piRNAKey words
Apis mellifera ligustica/midgut/ovary/testis/piRNA分类
生物学引用本文复制引用
张艺琼,陈大福,严提珍,郭睿,那志豪,李琪明,王梦怡,李婧娴,代梦远,邱剑丰,张荣华,卢兆辉..意大利蜜蜂piR-ame-1186994的靶基因与功能分析[J].昆虫学报,2025,68(3):260-270,11.基金项目
国家自然科学基金面上项目(32372943,32172792) （32372943,32172792）
国家现代农业产业技术体系专项资金(CARS-44-KXJ7) （CARS-44-KXJ7）
福建省自然科学基金面上项目(2022J01131334) （2022J01131334）
福建农林大学硕士生导帅团队项目(郭睿) （郭睿）
福建农林大学科技创新专项基金(KFb22060XA) （KFb22060XA）
福建省大学生创新创业训练计划项目(S202410389057,S202410389061) （S202410389057,S202410389061）
