p38 MAPK基因在西方蜜蜂越冬期 表达模式的研究
Expression of the p38 MAPK gene in different overwintering stages of Apis mellifera
徐 凯 杜亚丽 龙登隆 刘玉玲 陈东海 王 志 牛庆生
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DOI：10.7679/j.issn.2095-1353.2021.109
作者单位：吉林省养蜂科学研究所，吉林 132108；吉林省养蜂科学研究所，吉林 132108；吉林省养蜂科学研究所，吉林 132108；吉林省养蜂科学研究所，吉林 132108；吉林省养蜂科学研究所，吉林 132108；吉林省养蜂科学研究所，吉林 132108；吉林省养蜂科学研究所，吉林 132108
中文关键词：西方蜜蜂；越冬期；p38 MAPK；基因表达；抗寒性
英文关键词：Apis mellifera; overwintering stage; p38 MAPK; gene expression; cold resistance
中文摘要：
【目的】 p38 MAPK基因在昆虫的低温响应机制中发挥着重要作用，本研究旨在探究p38 MAPK基因在西方蜜蜂Apis mellifera越冬期内表达规律。【方法】 本研究对西方蜜蜂p38 MAPK蛋白序列进行生物信息学分析，并利用荧光定量PCR技术检测该基因在意大利蜜蜂Apis mellifera ligustica、欧洲黑蜂Apis mellifera mellifera、高加索蜂Apis mellifera caucasica和卡尼鄂拉蜂 Apis mellifera carnica于不同越冬时期和不同越冬方式下体内的表达量。【结果】 西方蜜蜂p38 MAPK包含1 083 bp的开放阅读框区域，编码360个氨基酸，包含TGY双磷酸化三肽模体序列和磷酸化激活环序列。p38 MAPK在蜜蜂所有组织中均有表达，分别在胸部和腹部处于最高和最低表达丰度。p38 MAPK mRNA在不同蜂种不同越冬时期的表达量存在显著差异，4个蜂种于室外越冬时p38 MAPK的表达量均显著高于室内越冬（P<0.05）；随着蜜蜂越冬持续时间的延长，意大利蜜蜂和欧洲黑蜂不同越冬方式下体内p38 MAPK的表达均呈现先上升后下降的表达趋势，高加索蜂在室外与室内越冬时体内该基因的表达量均表现出持续上升的表达趋势，该基因在卡尼鄂拉蜂中的表达趋势与其他3个蜂种呈现一定差异，卡尼鄂拉蜂p38 MAPK在室内越冬方式下表达量保持恒定，但在室外越冬方式下表现出先下降后上升的表达趋势；比较室外越冬情况下4个蜂种于不同越冬时期体内p38 MAPK表达量后发现，除在次年的1月外，其他越冬时期4个蜂种体内p38 MAPK的表达量均存在显著差异（P<0.05）。【结论】 p38 MAPK基因在西方蜜蜂越冬期内发挥着重要的生理功能，p38 MAPK信号通路可作为蜜蜂抗寒机制研究的候选信号通路。
英文摘要：
[Objectives] To investigate the expression of the p38 MAPK gene, which plays an important role in the insect cold response mechanism, in different overwintering stages of the western honeybee Apis mellifera. [Methods] The amino acid sequence of the Apis mellifera p38 MAPK gene was analyzed and its mRNA expression in different overwinter stages of Apis mellifera ligustica, Apis mellifera mellifera, Apis mellifera caucasica and Apis mellifera carnica detected using RT-qPCR. [Results] The p38 MAPK gene of A. mellifera includes a 1 083 bp open reading frame, encodes a polypeptide with 360 amino acids and contains a putative dual phosphorylation motif TGY and a phosphorylated activating ring sequence. P38 MAPK was expressed in all honeybee tissues. Highest and lowest expression occurred in the thorax and abdomen, respectively. Expression varied in different overwintering stages and subspecies. Expression was higher outdoors than indoors in all four subspecies P<0.05). In winter, p38 MAPK mRNA expression first increased then decreased in A. mellifera ligustica and A. mellifera mellifera which have different overwintering strategies. Expression increased in both outdoor and indoor overwintering colonies of A. mellifera caucasica. Unlike these three subspecies, expression remained constant in indoor overwintering colonies of A. mellifera carnica and tended to decrease after an initial increase in outdoor overwintering colonies of this subspecies. Overwintering expression levels varied among subspecies except in January of the second year (P<0.05). [Conclusion] The p38 MAPK gene plays an important physiological role in the overwintering processes of the western honeybee. The p38 MAPK signal pathway could be a candidate signal pathway for further study of the honeybee cold resistance mechanism.