意大利蝗Transformer 2基因鉴定及表达分析
Isolation and expression of the Calliptamus italicus Transformer 2 gene
张韦萱** 蒋思涵 扈鸿霞 赵 娜 季 荣 叶小芳***
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DOI：10.7679/j.issn.2095-1353.2025.036
作者单位：新疆师范大学生命科学学院，中亚区域跨境有害生物联合控制国际研究中心，新疆特殊环境物种保护与调控生物学实验室， 新疆特殊环境物种多样性应用与调控重点实验室，乌鲁木齐 830054
中文关键词：意大利蝗；Tra2；结构特征；表达模式；性别决定
英文关键词：Calliptamus italicus; Tra2; structure characteristics; expression pattern; sex determination
中文摘要：
【目的】 Transformer 2（Tra2）基因作为多种昆虫的性别决定关键基因，与Tra协同调控下游Dsx基因进行性特异性剪接，从而控制雌雄分化。本研究旨在鉴定意大利蝗Calliptamus
italicus 的Tra2基因结构特征及其在不同发育时期的表达模式。【方法】 基于意大利蝗卵的转录组数据库以及同源性比对结果，克隆了Tra2的开放阅读框（Open reading frame，ORF）序列片段，并进行结构预测、多重序列比对以及进化树分析；采用qRT-PCR技术检测意大利蝗卵期、不同发育龄期及成虫精巢和卵巢的Tra2表达量。【结果】 意大利蝗Tra2基因ORF序列含672 bp，编码223个氨基酸，Tra2编码的蛋白具有RRM-TRA结构域及特有Linker区。系统进化关系表明，意大利蝗Tra2编码的蛋白与鳞翅目亲缘关系最接近。Tra2在意大利蝗不同发育时期均有表达，在蝗卵发育过程中，阶段Ⅰ与阶段Ⅷ表达量较高；蝗蝻与成虫的Tra2表达量均为雌性显著高于雄性（P<0.05）。【结论】 明确了意大利蝗的Tra2基因结构特征，且雌虫Tra2的表达水平显著高于雄性，说明该基因对雌虫性器官的发育影响大于雄虫，为深入阐明Tra2在意大利蝗性别决定过程的调控作用奠定了基础。
英文摘要：
[Aim] To identify structural characteristics of the Transformer
2 (Tra2) gene,
a key gene involved in sex determination in many
insects, and quantify
its expression in different developmental stages of Calliptamus italicus. [Methods] Based on the transcriptome
database of C. italicus eggs and a comparison of homologous genes in
other species, the open reading frame (ORF) sequence fragment of Tra2 was cloned. The predicted structure, multiple sequence alignment and a
phylogenetic tree of the cloned sequence were obtained using bioinformatic
software. qRT-PCR was used to detect Tra2 expression in eggs, nymphs, and in the testes and ovaries of adult C. italicus. [Results] The ORF sequence of the Tra2 gene contains 672 bp
and encodes 223 amino acids. The protein encoded by Tra2 has an RRM-TRA
domain and a unique linker region. A phylogenetic tree indicates that the
protein encoded by Tra2 in C. italicus is most closely related to
homologous lepidopteran proteins. Tra2 was expressed in different
developmental stages of C. italicus but expression was the highest
during stage І and Ⅷ of egg development. Tra2 expression was significantly higher in females than in males in both nymphs and
adults (P<0.05). [Conclusion] Tra2 in C. italicus is closely related to homologous genes
in other lepidopteran species. Expression in females is significantly higher
than in males, suggesting that this gene has greater influence on the
development of female than male sexual organs. These results lay a foundation
for further elucidating the role of Tra2 in sex determination in C.
italicus.