昆虫学报 ›› 2023, Vol. 66 ›› Issue (7): 896-908.doi: 10.16380/j.kcxb.2023.07.004

• 研究论文 • 上一篇    下一篇

西方蜜蜂采集蜂咽下腺中高表达基因的筛选及表达分析

张娟1, 高艳1,2, 王若虹1, 李秋方1,3, 杨尚宁1, 周诗文1石丹丹1, 苏松坤1, *, 聂红毅1, *   

  1. (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; 3. 恩施土家族苗族自治州农业科学院, 恩施 445000)
  • 出版日期:2023-07-20 发布日期:2023-08-17

Screening and expression profiling of highly expressed genes in the hypopharyngeal glands of Apis mellifera (Hymenoptera: Apidae) foragers

ZHANG Juan1, GAO Yan1,2, WANG Ruo-Hong1, LI Qiu-Fang1, 3, YANG Shang-Ning1, ZHOU Shi-Wen1, SHI Dan-Dan1, SU Song-Kun1,*, NIE Hong-Yi1,*    

  1. (1. College of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, China; 2. Jiangxi Apiculture Research Institute (Jiangxi Apiculture Technology Extension Station), Nanchang 330052, China; 3. Enshi Tujia and Miao Autonomous Prefecture Academy of Agricultural Sciences, Enshi 445000, China)
  • Online:2023-07-20 Published:2023-08-17

摘要: 【目的】本研究旨在筛选西方蜜蜂Apis mellifera采集蜂咽下腺中高表达的基因,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】基于前期已获得西方蜜蜂3日龄工蜂(3 d_W)、10日龄哺育蜂(10 d_N)、10日龄采集蜂(10 d_F)、21日龄哺育蜂(21 d_N)和21日龄采集蜂(21 d_F)的咽下腺转录组数据,筛选咽下腺差异表达基因(differentially expressed genes, DEGs),并对10日龄和21日龄采集蜂咽下腺中高表达的DEGs进行GO和KEGG富集分析;利用qPCR检测西方蜜蜂采集蜂咽下腺中高表达的4个DEGs (LOC409889, LOC406114, LOC408453和LOC100578735)在不同发育时期(卵、幼虫、蛹、刚出房工蜂、10日龄哺育蜂和21日龄采集蜂)和21日龄采集蜂组织(腹、胸、触角、上颚腺、咽下腺、蛰针、足、肠道、翅和脑)中的表达量。【结果】比较转录组结果表明,164个上调表达的DEGs在21日龄采集蜂咽下腺中的表达量明显高于在3日龄工蜂、10日龄哺育蜂和21日龄哺育蜂咽下腺中的表达量,且在10日龄采集蜂咽下腺中的表达量也高于在10日龄哺育蜂咽下腺中的表达量;105个下调表达的DEGs的表达趋势与上述164个DEGs的相反。GO分析结果表明,10日龄和21日龄采集蜂咽下腺中高表达的164个DEGs显著富集于硫酸盐跨膜转运蛋白活性、硫化合物跨膜转运蛋白活性、铁离子结合和氧化还原酶活性;下调表达的105个DEGs显著富集于核糖体、细胞内核糖核蛋白复合物和核糖核蛋白复合物;KEGG通路分析结果表明,105个下调表达的DEGs显著富集于核糖体以及淀粉和蔗糖代谢两个通路。qPCR检测结果显示,LOC409889, LOC406114, LOC408453和LOC100578735均在21日龄采集蜂中表达量最高;LOC406114和LOC100578735在21日龄采集蜂咽下腺中的表达量最高,而在其他组织中表达量较低或不表达。【结论】本研究通过比较转录组筛选鉴定了西方蜜蜂采集蜂咽下腺中高表达的DEGs,为深入研究采集蜂咽下腺的功能提供了数据,同时也为研究西方蜜蜂的采集行为及采集力强蜂种的分子选育提供新的视角。

关键词: 西方蜜蜂, 咽下腺, 差异表达基因, α-淀粉酶, 丙酮酸脱氢酶α亚基E1组分

Abstract: 【Aim】 This study aims to screen genes highly expressed in the hypopharyngeal glands (HGs) of Apis mellefera foragers, and to provide a basis for further studying the genes related to honey bee foraging behavior.【Methods】 Differentially expressed genes (DEGs) in the HGs were screened based on the previously sequenced HGs transcriptome data of the 3-day-old workers (3 d_W), 10-day-old nurses (10 d_N), 10-day-old foragers (10 d_F), 21-day-old nurses (21 d_N), 21day-old foragers (21 d_F) and the DEGs highly expressed in the HGs of the 10-day-old foragers and 21-day-old foragers of A. mellefera were subjected to GO and KEGG enrichment analysis. Meanwhile, the expression levels of four highly expressed DEGs (LOC409889, LOC406114, LOC408453 and LOC100578735) in the HGs of foragers in different developmental stages (egg, larva, pupa, newly emerged workers, 10-day-old nurses and 21-day-old foragers) and tissues (abdomen, thorax, antenna, mandibular gland, hypoharyngeal gland, sting, leg, gut, wing and brain) of the 21-day-old foragers of A. mellefera were examined using qPCR.【Results】Comparative transcriptome result showed that the expression levels of 164 up-regulated DEGs in the HGs of the 21-day-old foragers were obviously increased as compared with those of the 3-day-old workers, 10-day-old nurses and 21-day-old nurses, and those of the 10-day-old foragers were also higher than those of the 10-day-old nurses. While the expression trend of the 105 down-regulated DEGs was opposite with that of the above 164 DEGs. GO analysis result showed that the 164 highly expressed DEGs in the HGs of the 10-/21-day-old foragers were enriched significantly in sulfate transmembrane transporter activity, sulfur compound transmembrane transporter activity, iron ion binding and oxidoreductase activity, and the 105 down-regulated DEGs were enriched significantly in ribosome, intracellular ribonucleoprotein complex and ribonucleoprotein complex. KEGG pathway analysis result revealed that the 105 down-regulated DEGs were significantly enriched in two pathways ribosome, and starch and sucrose metabolism. qPCR detection result indicated that the expression levels of LOC409889, LOC406114, LOC408453 and LOC100578735 were the highest in the 21-day-old foragers, and those of LOC406114 and LOC100578735 were the highest in the HGs of the 21-day-old foragers and showed low expression level or no expression in other tissues.【Conclusion】 The highly expressed DEGs identified in the HGs of A. mellifera forager were screened in this study via comparative transcriptome, which provides a valuable source of data for the function study of forager HGs, and also provides a new perspective for further studying the foraging behavior of A. mellefera and the molecular breeding selection of bee species with strong foraging ability.

Key words: Apis mellifera, hypopharyngeal glands, differentially expressed genes, alpha-amylase, pyruvate dehydrogenase E1 component subunit alpha