异色瓢虫酪氨酸羟化酶基因不同发育历期表达及RNAi抑制黑化效果分析  

Expression of tyrosine hydroxylase gene in development stage and effect of RNAi on suppressing melanin in Harmonia axyridis

作　　者：张道伟[1] 王莎莎[2] 张萌 钱正敏[1] 曾伯平[1] 唐斌 ZHANG Dao-Wei;WANG Sha-Sha;ZHANG Meng;QIAN Zheng-Min;ZENG Bo-Ping;TANG Bin(College of Biology and Agriculture,Zunyi Normal University,Zunyi 563006,Guizhou Province,China;College of Life and Environmental Science,Hangzhou Normal University,Hangzhou 310036,China)

机构地区：[1]遵义师范学院生物与农业科技学院,贵州遵义563006 [2]杭州师范大学生命与环境科学学院,杭州310036

出　　处：《环境昆虫学报》2019年第2期259-266,共8页Journal of Environmental Entomology

基　　金：国家重点研发计划专项"天敌昆虫防控技术及产品研发"项目(2017YFD0201000);杭州市农业/社会发展主动设计项目(20190101A01);贵州省科学技术基金(黔科合JZ字[2014]2014号)

摘　　要：酪氨酸代谢在昆虫黑色素形成中具有重要的调控作用。本研究为探讨酪氨酸代谢调控异色瓢虫黑色素代谢平衡及其发育的影响,先检测异色瓢虫不同发育阶段酪氨酸羟化酶(Tyrosine hydroxylase,TH)基因表达情况,采用RNAi技术对4龄幼虫注射dsTH RNA抑制酪氨酸羟化酶mRNA表达,探索其在蛹期和成虫羽化阶段黑色素形成中的潜在功能。研究结果表明,TH基因在4龄幼虫体内的表达量最高,而在预蛹阶段和羽化后第2天开始表达量显著性下降。其次,dsTH RNA注射后能够显著降低TH本基因的表达,且异色瓢虫4龄幼虫发育缩短,与注射dsGFP RNA组相比较,提前化蛹。最后,dsTH RNA注射后异色瓢虫蛹的重量略有增加,且发育出现异常,即蛹的颜色变化不加深,与对照组相比较,无黑色斑点出现。这些结果表明成虫是在蛹期开始形成黑色素的,及酪氨酸羟化酶能够通过调控酪氨酸代谢途径而影响异色瓢虫化蛹阶段的发育和表皮颜色的变化。相关研究结果有助于丰富酪氨酸代谢调控异色瓢虫表型的内容,为将来保护异色瓢虫,并将其用于害虫生物防治提供理论基础。Tyrosine metabolism plays an important regulatory role in the formation of insect melanin. This study was to investigate the effect of regulation of tyrosine metabolism on melanin metabolism and insect s development. Firstly, we detected the expression of tyrosine hydroxylase ( TH ) gene in different developmental stages of Harmonia axyridis , and then used RNAi technology to inhibit the expression of tyrosine hydroxylase mRNA by injecting dsTH RNA into the fourth instar larvae. The above results were used to explore the potential function of TH in melanin formation during pupa and adults after eclosion stages. The results showed that the expression of TH gene was highest in the fourth instar larvae, and then significantly decreased on the prepupa stage and on the second day after eclosion. Secondly, results have shown that dsTH RNA can significantly reduce the expression of TH gene after injection. The developmental time of the fourth instar larvae of H. axyridis was shortened after dsTH RNA injection, and larvae were prematurely sputum compared with the dsGFP RNA injected group. Finally, after dsTH RNA injection, we found that the weight of pupa increased slightly, and the development of pupa was abnormal. Compared with the control group, the color of the pupa remained golden yellow and no black spots were formed on the surface. These results indicate that adults begin to form melanin during the stage of pupa, and tyrosine hydroxylase can affect the development of the pupa stage and the change of epidermal color by regulating the tyrosine metabolic pathway. Our findings will help to enrich the tyrosine metabolism and regulate the phenotype of H. axyridis , and provide a theoretical basis for the protection of H. axyridis in the future and its application to pest biological control.

关 键 词：异色瓢虫 酪氨酸羟化酶 RNA干扰 酪氨酸代谢 黑色素合成 

分 类 号：Q963[生物学—昆虫学] S476[农业科学—农业昆虫与害虫防治]