Characterization of two juvenile hormone epoxide hydrolases by RNA interference in the Colorado potato beetle
Characterization of two juvenile hormone epoxide hydrolases by RNA interference in the Colorado potato beetle
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通过 RNA 干扰表征科罗拉多马铃薯甲虫的两种保幼激素环氧化物水解酶
DOI：
10.1016/j.gene.2015.06.032
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发表时间：
2015-10-10
期刊：
影响因子：
3.5
通讯作者：
Li, Guo-Qing
中科院分区：
文献类型：
--
作者：
Lu, Feng-Gong;Fu, Kai-Yun;Li, Guo-Qing
在昆虫的不同发育阶段，幼体激素OH)的滴度通过合成和降解途径受到严格调控。在JH降解过程中，JH环氧化物水解酶（JHEH）将JH转化为JH二醇，并将JH酸水解为JH酸二醇。本研究从Leptinotarsa decemlineata中克隆了两个全长LdJHEH cdna，暂时命名为LdJHEH1和LdJHEH2。在第2天4龄幼虫的胸肌、脑-心-体复合体、前肠、中肠、后肠、腹侧神经节、马尔比氏小管、脂肪体、表皮和血细胞以及成虫的雌性卵巢和雄性生殖器官中均检测到这两种mrna。LdJHEH1和LdJHEH2在幼虫的整个生命周期中均有表达，并在最后（4龄）幼虫蜕化32 h后达到峰值。构建了4个分别靶向LdJHEH1和LdJHEH2的双链rna (dsRNAs) （d5JHEH1-1、dsJHEH1-2、ds1HEH2-1、dsJHEH2-2）并进行了细菌表达。摄入dsJHEH1-1、dsJHEH1-2、dsJHEH2-1、dsJHEH2-2以及dsJHEH1-1 + d5JHEH2-1的混合物，成功敲低了相应的靶基因功能，显著提高了JH滴度，上调了Kriippel同源物1 (LdKr-h1) mRNA水平。敲低LdJHEH1或LdJHEH2，或敲低两个基因中的任何一个，都能轻微降低幼虫体重，延缓幼虫发育，并显著损害成虫羽化。因此，LdJHEH1和LdJHEH2的敲低影响了科罗拉多马铃薯甲虫对JH的降解。(C) 2015 Elsevier B.V. Al！版权。
In insect, juvenile hormone OH) titers are tightly regulated in different development stages through synthesis and degradation pathways. During JH degradation, JH epoxide hydrolase (JHEH) converts JH to JH diol, and hydrolyses JH acid to JH acid diol. In this study, two full length LdJHEH cDNAs were cloned from Leptinotarsa decemlineata, and were provisionally designated LdJHEH1 and LdJHEH2. Both mRNAs were detectable in the thoracic muscles, brain-corpora cardiaca-corpora allata complex, foregut, midgut, hindgut, ventral ganglia, Malpighian tubules, fat bodies, epidermis, and hemocytes of the day 2 fourth-instar larvae, and in female ovaries as well as male reproductive organs of the adults. Moreover, both LdJHEH1 and LdJHEH2 were expressed throughout all larval life, with the highest peaks occurring 32 h after ecdysis of the final (fourth) instar larvae. Four double-stranded RNAs (dsRNAs) (d5JHEH1-1, dsJHEH1-2, ds1HEH2-1, dsJHEH2-2) respectively targeting LdJHEH1 and LdJHEH2 were constructed and bacterially expressed. Ingestion of dsJHEH1-1, dsJHEH1-2, dsJHEH2-1, dsJHEH2-2, and a mixture of dsJHEH1-1 + d5JHEH2-1 successfully knocked down corresponding target gene function, and significantly increased JH titer and upregulated Kriippel homolog 1 (LdKr-h1) mRNA level. Knockdown of either LdJHEH1 or LdJHEH2, or both genes slightly reduced larval weight and delayed larval development, and significantly impaired adult emergence. Therefore, it is suggested that knockdown LdJHEH1 and LdJHEH2 affected JH degradation in the Colorado potato beetle. (C) 2015 Elsevier B.V. Al! rights reserved.