ⅰ群多糖单加氧酶对两种森林害虫甲虫换羽过程中几丁质角质层周转的需求。

IF 2.3 2区 农林科学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Daehyeong Kim, Subbaratnam Muthukrishnan, Karl J Kramer, Yasuyuki Arakane, Mi Young Noh
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引用次数: 0

摘要

多糖单加氧酶(LPMOs)是一种广泛存在于生物体内的酶,它能够氧化裂解包括几丁质和纤维素在内的晶体多糖中的糖苷键。昆虫LPMOs属于辅助活性家族15 (LPMO15/AA15),并被进一步划分为至少四个亚群。然而,与来自微生物和病毒的LPMOs不同,它们在昆虫中的生理功能尚未得到很好的研究。在本研究中,我们研究了两组I型LPMO15s, MaLPMO15-1和PhLPMO15-1在两种重要的森林害虫长角甲虫——日本松索甲虫(Monochamus alternatus)和黄斑长角甲虫(Psacothea hilaris)换壳过程中几质角质层更新中的功能。Real-time qPCR显示MaLPMO15-1和PhLPMO15-1在发育后期的表达模式相似,在幼蛹和幼蛹阶段表达水平较高,此后表达水平下降。将MaLPMO15-1 (dsMaLPMO15-1)或PhLPMO15-1 (dsPhLPMO15-1)的双链RNA (dsRNA)分别注射到交替斑胸蚜和黄斑胸蚜的末龄幼虫中,对后续的幼虫-蛹换羽没有影响,得到的蛹发育正常。然而,蛹无法接近成虫期,被困在旧蛹的角质层中死亡。透射电镜分析显示,与增强的绿色荧光蛋白(dsEGFP)处理的对照昆虫的dsRNA不同,dsMaLPMO15-1-和dsphlpmo15 -1处理的成虫蛹表皮的内质层都没有降解,其中水平几质层基本保持完整。这些结果表明,I组LPMO15-1酶在蛹角质层几丁质转换中起作用,这对成虫蜕皮至关重要。由于LPMO15-1在许多昆虫物种中高度保守,因此该基因/酶是控制交替稻蠹蛾和小叶螟以及其他害虫物种种群的潜在靶点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Requirement of group I lytic polysaccharide monooxygenase for turnover of chitinous cuticle during moulting in two forest pest beetles, Monochamus alternatus and Psacothea hilaris.

Lytic polysaccharide monooxygenases (LPMOs) that are capable of oxidative cleavage of glycosidic bonds in crystalline polysaccharides including chitin and cellulose are widely distributed among organisms. Insect LPMOs belong to auxiliary activity family 15 (LPMO15/AA15) and have been classified further into at least four subgroups. However, unlike LPMOs from microorganisms and viruses, their physiological functions in insects have not been well studied. In the present work, we investigated the functions of two group I LPMO15s, MaLPMO15-1 and PhLPMO15-1, in chitinous cuticle turnover during moulting of two important forest pest longhorn beetles-the Japanese pine sawyer beetle, Monochamus alternatus, and the yellow spotted longicorn beetle, Psacothea hilaris. Real-time qPCR showed a similar pattern of expression of MaLPMO15-1 and PhLPMO15-1 during late stages of development with high levels present at young pharate pupal and young pupal stages and declining thereafter. Injection of double-stranded RNA (dsRNA) for MaLPMO15-1 (dsMaLPMO15-1) or PhLPMO15-1 (dsPhLPMO15-1) into last instar larvae of M. alternatus and P. hilaris, respectively, did not affect subsequent larval-pupal moulting and the resulting pupae exhibited normal development. However, the pupae were unable to eclose to the adult stage and died entrapped inside their old pupal cuticle. TEM analysis revealed that, unlike the respective dsRNA for the enhanced green fluorescent protein (dsEGFP)-treated control insects, both dsMaLPMO15-1- and dsPhLPMO15-1-treated pharate adults failed to degrade the endocuticular layer of their pupal cuticle in which the horizontal chitinous laminae remained largely intact. These results demonstrate that the group I LPMO15-1 enzymes play a role in pupal cuticle chitin turnover, which is critical for moulting to the adult. Because LPMO15-1 is highly conserved among many insect species, this gene/enzyme is a potential target for the control of populations of both M. alternatus and P. hilaris as well as other pest insect species.

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来源期刊
Insect Molecular Biology
Insect Molecular Biology 生物-昆虫学
CiteScore
4.80
自引率
3.80%
发文量
68
审稿时长
6-12 weeks
期刊介绍: Insect Molecular Biology has been dedicated to providing researchers with the opportunity to publish high quality original research on topics broadly related to insect molecular biology since 1992. IMB is particularly interested in publishing research in insect genomics/genes and proteomics/proteins. This includes research related to: • insect gene structure • control of gene expression • localisation and function/activity of proteins • interactions of proteins and ligands/substrates • effect of mutations on gene/protein function • evolution of insect genes/genomes, especially where principles relevant to insects in general are established • molecular population genetics where data are used to identify genes (or regions of genomes) involved in specific adaptations • gene mapping using molecular tools • molecular interactions of insects with microorganisms including Wolbachia, symbionts and viruses or other pathogens transmitted by insects Papers can include large data sets e.g.from micro-array or proteomic experiments or analyses of genome sequences done in silico (subject to the data being placed in the context of hypothesis testing).
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