CHARACTERIZATION AND FUNCTIONAL STUDY OF A PUTATIVE JUVENILE HORMONE DIOL KINASE IN THE COLORADO POTATO BEETLE Leptinotarsa decemlineata (Say).
CHARACTERIZATION AND FUNCTIONAL STUDY OF A PUTATIVE JUVENILE HORMONE DIOL KINASE IN THE COLORADO POTATO BEETLE Leptinotarsa decemlineata (Say).
复制标题
DOI：
10.1002/arch.21251
复制
发表时间：
2015-08
影响因子：
2.2
通讯作者：
Kaiyun Fu;Feng-gong Lü;Wenchao Guo;Guo-Qing Li
中科院分区：
文献类型：
--
作者：
Kaiyun Fu;Feng-gong Lü;Wenchao Guo;Guo-Qing Li
保幼激素二醇激酶(JHDK)是一种参与JH降解的酶。本文从科罗拉多州马铃薯甲虫Leptinotarsa decemlineata中克隆了一个可能的JHDK基因(LdJHDK)。该基因全长814bp，含有555bp的开放阅读框，编码184个氨基酸的蛋白质。LdJHDK与以前报道的昆虫JHDK具有高度的同源性。它具有三个保守的嘌呤核苷酸结合元件，并含有三个EF-Hand基序(螺旋-环-螺旋结构域)。LdJHDK基因主要在后肠和马氏管中表达。此外，在胸肌、脑-心体-翼体复合体、前肠、中肠、腹神经节、脂肪体、表皮和血细胞中也有微量的LdJHDK基因表达。此外，LdJHDK在所有发育阶段都有表达。在1、2、3龄幼虫中，LdJHDK在蜕皮前和蜕皮后表达水平较高，而在中龄期表达水平较低。在4龄幼虫中，LdJHDK峰值出现在蜕皮后56h。摄取针对LdJHDK的双链RNA(DsRNA)成功地下调了靶基因，提高了JH滴度，并显著上调了LdKr-H1的mRNA水平。敲除LdJHDK基因显著影响成虫羽化。因此，我们在十字乳杆菌中提供了一系列实验证据来支持LdJHDK编码参与JH降解的功能蛋白。
Juvenile hormone diol kinase (JHDK) is an enzyme involved in JH degradation. In the present article, a putative JHDK cDNA (LdJHDK) was cloned from the Colorado potato beetle Leptinotarsa decemlineata. The cDNA consists of 814 bp, containing a 555 bp open reading frame encoding a 184 amino acid protein. LdJHDK reveals a high degree of identity to the previously reported insect JHDKs. It possesses three conserved purine nucleotide-binding elements, and contains three EF-hand motifs (helix-loop-helix structural domains). LdJHDK mRNA was mainly detected in hindgut and Malpighian tubules. Besides, a trace amount of LdJHDK mRNA was also found in thoracic muscles, brain-corpora cardiaca-corpora allata complex, foregut, midgut, ventral ganglia, fat body, epidermis, and hemocytes. Moreover, LdJHDK was expressed throughout all developmental stages. Within the first, second, and third larval instar, the expression levels of LdJHDK were higher just before and right after the molt, and were lower in the intermediate instar. In the fourth larval instar, the highest peak of LdJHDK occurred 56 h after ecdysis. Ingestion of double-stranded RNA (dsRNA) against LdJHDK successfully knocked down the target gene, increased JH titer, and significantly upregulated LdKr-h1 mRNA level. Knockdown of LdJHDK significantly impaired adult emergence. Thus, we provide a line of experimental evidence in L. decemlineata to support that LdJHDK encodes function protein involved in JH degradation.