关键词: 意大利蜜蜂, 铁蛋白, 基因克隆, 温度胁迫, 表达分析

Abstract: 【Aim】 This study aims to clone and identify the ferritin gene AmFer3HCH from Apis mellifera ligustica, to analyze its tissue expression changes in response to temperature stress, and to explore the role of ferritin in encountering the extreme temperature stress. 【Methods】 The cDNA sequence of AmFer3HCH gene was amplified from A. m. ligustica by PCR, and its amino acid sequence was identified by bioinformatic tools. The expression levels of this gene in the thorax, midgut, and fat body of A. m. ligustica worker adults under temperature stress (40℃ and 5℃) were detected by real-time quantitative PCR. 【Results】 A ferritin gene named AmFer3HCH was cloned from A. m. ligustica by PCR and its cDNA fragment is 507 bp in length. Its nucleotide sequence has 100% identity to that of the Fer3HCH gene (GenBank accession no.: GB55416) in A. m. ligustica genome in GenBank database. AmFer3HCH was identified as a ferritin heavychain subunit and clustered in one phyletic clade with other heavy-chain ferritins. The amino acid sequence of AmFer3HCH has a typical ferroxidase activity center, ferrihydrite nucleation center and iron channel as in most ferritin family members. Real-time quantitative PCR analysis showed that the expression levels of AmFer3HCH in the thorax, midgut and fat body of A. m. ligustica worker adults after brief exposure to high temperature stress (40℃ for 0-48 h) presented a tendency of upregulation first, then downregulation and upregulation again as compared to the control group. However, after the adults were exposed to brief low temperature stress (5℃ for 0-6 h), the expression level of AmFer3HCH was up-regulated first, and then down-regulated in the midgut and fat body, but showed a falling tendency in the thorax. 【Conclusion】 AmFer3HCH is a heavy-chain ferritin and plays an important role in response to extreme temperature stress in A. m. ligustica.

Key words: Apis mellifera ligustica, ferritin, gene cloning, temperature stress, expression analysis