西方蜜蜂采集蜂上颚腺中高表达基因的筛选与表达分析  

Screening and expression profiling of highly expressed genes in the mandibular glands of Apis mellifera foragers

作　　者：李秋方 高艳 梁立强 李正汉卿 朱雅楠 张娟 杨尚宁 傅云熙 苏松坤 聂红毅 LI Qiu-Fang;GAO Yan;LIANG Li-Qiang;LI Zheng-Han-Qing;ZHU Ya-Nan;ZHANG Juan;YANG Shang-Ning;FU Yun-Xi;SU Song-Kun;NIE Hong-Yi(College of Animal Sciences(College of Bee Science),Fujian Agriculture and Forestry University,Fuzhou 350002,China;Jiangxi Apiculture Technology Extension Station,Jiangxi Apiculture Research Institute,Nanchang 330052,China)

机构地区：[1]福建农林大学动物科学学院(蜂学学院),福州350002 [2]江西省养蜂研究所,江西省蜂业技术推广站,南昌330052

出　　处：《昆虫学报》2022年第8期937-948,共12页Acta Entomologica Sinica

基　　金：国家自然科学基金面上项目(31772684);财政部和农业农村部:国家现代农业产业技术体系(CARS-44)。

摘　　要：【目的】本研究旨在筛选西方蜜蜂Apis mellifera采集蜂上颚腺中高表达基因,为进一步筛选和研究蜜蜂采集行为相关基因提供依据。【方法】基于前期测序的西方蜜蜂5种不同职能工蜂(3日龄工蜂、10日龄哺育蜂、10日龄采集蜂、21日龄哺育蜂和21日龄采集蜂)上颚腺转录组数据,筛选采集蜂上颚腺的差异表达基因(differentially expressed genes,DEGs),并对这些DEGs进行GO和KEGG分析;qRT-PCR检测随机选取的8个DEGs在10日龄哺育蜂和10日龄采集蜂上颚腺以及两个关键DEGs(Δ-1-吡咯啉-5-羧酸合成酶基因Amp5cs和细胞色素P4509e2基因CYP9Q 3)在工蜂不同发育时期和采集蜂各组织中的表达量。【结果】筛选到22个DEGs在21日龄采集蜂上颚腺中的表达量显著高于在3日龄工蜂、10日龄哺育蜂和21日龄哺育蜂上颚腺中的表达量,同时在10日龄采集蜂上颚腺中的表达量也显著高于在10日龄哺育蜂上颚腺中的表达量。GO和KEGG富集分析显示这些DEGs主要富集在胆固醇代谢、半乳糖代谢、淀粉和蔗糖代谢、精氨酸和脯氨酸代谢、细胞凋亡-果蝇、氨基酸生物合成等方面。qRT-PCR结果表明,8个DEGs(LOC100576395,LOC411983,LOC410235,LOC725581,LOC410527,LOC406131,LOC408453和LOC410253)的表达模式与转录组数据的表达模式一致;2个关键DEGs Amp5cs和CYP9Q 3在工蜂各发育阶段均有表达,且在采集蜂中表达量最高;Amp5cs在采集蜂腹、胸、上颚腺和触角中高量表达,P4509e 2在采集蜂触角和足中表达量显著高于在其他组织中的。【结论】本研究在减小日龄因素干扰下筛选了西方蜜蜂采集蜂上颚腺中22个高表达的DEGs,这些DEGs可能主要参与采集蜂上颚腺生理发育以及能量供应、外源性物质解毒、花蜜转化等代谢通路,进而影响蜜蜂的采集行为。这些结果为西方蜜蜂上颚腺的功能研究提供理论参考,同时也为采集力强的新品种培育奠定了基础。【Aim】This study aims to screen highly expressed genes in the mandibular glands of Apismellifera foragers,so as to provide the basis for further screening and study of genes related with the foraging behavior of honey bees.【Methods】Differentially expressed genes(DEGs)in the mandibular glands of foragers were screened based on previously sequenced transcriptome data of mandibular glands of five kinds of workers of A.mellifera with different tasks(3-day-old worker bees,10-day-old nurse bees,10-day-old forager bees,21-day-old nurse bees and 21-day-old forager bees),and GO and KEGG analyses of these DEGs were performed.qRT-PCR was used to detect the expression levels of eight randomly selected DEGs in the mandibular glands of 10-day-old nurse bees and 10-day-old forager bees,and two key DEGs includingΔ-1-pyrroline-5-carboxylate synthase gene Amp5cs and cytochrome P4509e2 gene CYP9 Q3 in different developmental stages of workers and tissues of foragers.【Results】The expression levels of 22 DEGs in the mandibular glands of 21-day-old foragers were significantly higher than those of 3-day-old workers,10-day-old nurses and 21-day-old nurses.Meanwhile,the expression levels of the above 22 DEGs in the mandibular glands of 10-day-old foragers of A.mellifera were also significantly higher than those of 10-day-old nurses.GO and KEGG enrichment showed that these DEGs were mainly enriched in cholesterol metabolism,galactose metabolism,starch and sucrose metabolism,arginine and proline metabolism,apoptosis-fly,and biosynthesis of amino acids.qRT-PCR results showed that the expression patterns of eight DEGs(LOC100576395,LOC411983,LOC410235,LOC725581,LOC410527,LOC406131,LOC408453 and LOC410253)were consistent with those of the transcriptome data.Two key DEGs Amp5cs and CYP9Q 3 were expressed in various developmental stages of workers,with the highest expression level in foragers.The expression levels of Amp5cs in abdomen,thorax,mandibular glands and antennae of foragers were higher,and those of CYP9Q 3 were significantl

关 键 词：西方蜜蜂 上颚腺 采集行为 Δ-1-吡咯啉-5-羧酸合成酶 细胞色素P4509e2 

分 类 号：Q965[生物学—昆虫学]