飞蝗Knickkopf家族基因表达对蜕皮激素的响应
The transcriptional response of Knickkopf family genes to 20-hydroxyecdysone in Locusta migratoria
张 睿；赵小明；刘晓健；张学尧；张建珍
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DOI：10.7679/j.issn.2095-1353.2022.067
作者单位：山西大学应用生物学研究所，太原 030006；山西大同大学生命科学学院，大同 037009
中文关键词：飞蝗；Knickkopf；蜕皮激素；RNA干扰
英文关键词：Locusta migratoria; Knickkopf; 20-hydroxyecdysone; RNAi
中文摘要：
【目的】 研究飞蝗Locusta migratoria Knickkopf（Knk）家族基因mRNA表达对蜕皮激素（20-hydroxyecdysone，20E）的响应情况，丰富飞蝗Knk家族基因功能研究，为飞蝗表皮发育相关基因的转录调控研究奠定基础。【方法】 采用RT-qPCR方法，对LmKnk家族4个基因在飞蝗不同发育天数（5龄若虫第1天到成虫第3天）的表皮中的表达特性进行分析；向飞蝗体腔注射20E，分析LmKnk家族4个基因表达的变化情况；采用RNAi技术干扰20E受体基因LmEcR，分析LmKnk家族基因的表达变化情况。【结果】 通过RT-qPCR检测，发现LmKnk在飞蝗5龄第1天至成虫第3天不同发育天数的表皮中均衡表达，而LmKnk2，LmKnk3-FL及LmKnk3-5¢ 基因均在蜕皮前表达量逐渐升高，蜕皮后迅速降低。向飞蝗体腔注射20E后，发现飞蝗Knickkopf家族4个基因对20E均有应答，但响应时间点有差异，LmKnk 和LmKnk2的表达量分别在注射20E 3 h和 6 h后开始升高，而LmKnk3-FL和LmKnk3-5¢的表达量在注射20E 12 h后开始上升。飞蝗5龄第1天若虫注射dsLmEcR后，发现Knickkopf家族4个基因表达均下调。【结论】 飞蝗Knickkopf家族基因的表达受20E调控，是20E的下游应答基因。
英文摘要：
[Objectives] To investigate the
transcriptional response of Knickkopf family genes to 20-hydroxyecdysone in Locusta migratoria in order to advance
research on the function of LmKnk family genes, and lay a foundation for
further research on the regulation of the transcription of genes involved in
cuticle development in L. migratoria. [Methods] RT-qPCR was used to analyze the expression
characteristics of four LmKnk genes in the integument of L.
migratoria from 1-day-old fifth-instar nymphs to 3-days-old adults. 20E was then injected into nymphs to analyze the expression of LmKnk family genes and RNAi was used
to suppress the expression of the 20E receptor gene LmEcR and thereby
detect transcriptional changes in LmKnk family genes. [Results] RT-qPCR revealed that LmKnk was evenly distributed in the integument of 1-day-old fifth-instar
nymphs to 3-days-old adults, and that the expression of LmKnk2, LmKnk3-FL and LmKnk3-5¢ all gradually increased before molting but
decreased rapidly after molting. Each of the LmKnk family genes
responded to 20E at a different time point. Expression of LmKnk and LmKnk2 rapidly increased 3 and 6 hours after 20E injection, respectively,
whereas expression of LmKnk3-FL and LmKnk3-5¢ was upregulated 12 hours after 20E injection. dsLmEcR injection reduced transcription of all four LmKnk family genes in
1-day-old fifth-instar nymphs. [Conclusion] LmKnk family genes are downstream genes that can be regulated by 20E in L. migratoria.