Cloning and RNAi-mediated functional characterization of MaLac2 of the pine sawyer, Monochamus alternatus

文献类型： 外文期刊

第一作者： Niu, B. -L.

作者： Niu, B. -L.;Shen, W. -F.;Liu, Y.;Weng, H. -B.;He, L. -H.;Meng, Z. -Q.;Tao, Y. -Z.;Mu, J. -J.;Wu, Z. -L.;Jiang, P.

作者机构：

关键词： Monochamus alternatus;cuticle sclerotization;MaLac2;RNA interference (RNAi);laccase 2

期刊名称：INSECT MOLECULAR BIOLOGY （ 影响因子：3.585； 五年影响因子：3.215 ）

ISSN： 0962-1075

年卷期： 2008 年 17 卷 3 期

页码：

收录情况： SCI

摘要： Laccase, a member of a group of proteins collectively known as multicopper oxidases, is hypothesized to play an important role in insect cuticle sclerotization by oxidizing catechols in the cuticle to their corresponding quinones, which then catalyze protein cross-linking reactions. Laccase 2 has been proved as the gene required for beetle cuticle tanning through RNA interference (RNAi) experiments on red flour beetle Tribolium castaneum. The pine sawyer beetle, Monochamus alternatus (Coleoptero: Cerambycidae) is the insect serving as a major vector of the pinewood nematode, Bursaphelenchus xylophilus, which is the causative agent for pine wilt disease. The cDNA of MaLac2 was cloned from the insect in this study. The conceptual amino-acid sequence deduced was much conserved with other known insect laccases, particularly with the enzyme of Tribolium castaneum. Injection in hemolymph of pine sawyer larva of dsRNA targeting the laccase 2 mRNA leads to important alterations of the tanning, hardening and sclerotization of the pupal and adult cuticles. Defaults appear in a dose-dependent manner and high loads of dsRNA are lethal. The decrease of the endogenous laccase 2 mRNA affects the procuticle which is thinner and without the characteristic piling up of successive layers. The observations reinforce the role of laccase 2 as an essential phenoloxidase for making cuticle.

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