Tang Jing-Wen, Wang Qi, Jiang Yun-Min, Jiang Yi-Ren, Wang Yong, Liu Wei
Liaoning Engineering and Technology Research Center for Insect Resources, College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang 110866, China.
Int J Mol Sci. 2024 Dec 31;26(1):296. doi: 10.3390/ijms26010296.
Chitin deacetylases (CDAs) are carbohydrate esterases associated with chitin metabolism and the conversion of chitin into chitosan. Studies have demonstrated that chitin deacetylation is essential for chitin organization and compactness and therefore influences the mechanical and permeability properties of chitinous structures, such as the peritrophic membrane (PM) and cuticle. In the present study, two genes ( and ) encoding CDA protein isoforms were identified and characterized in Chinese oak silkworm () larvae. Although five signature motifs were identified, CDA5 proteins only have the chitin-deacetylated catalytic domain. Spatiotemporal expression pattern analyses revealed that both transcripts presented the highest abundance in the anterior region of the midgut during the feeding period after molting, suggesting their role in chitin turnover and PM assembly. The down-regulation of and via RNA interference (RNAi) was correlated with the breakage of chitin microfibrils in the PM, suggesting that group V CDAs were essential for the growth and assembly of the chitinous layer. Additionally, and may have non-overlapping functions that regulate the morphological characteristics of PM chitin construction in different ways. Larvae injected with double-stranded RNA (dsRNA) against and transcripts were less resistant to infection by than those in the control groups. These results revealed that down-regulating and had independent effects on the PM structure and undermined the intactness of the PM, which disrupted the function of the PM against microsporidia infection per os. Our data provide new evidence for differentiating CDA functions among group V CDAs in lepidopteran insects.
几丁质脱乙酰酶(CDAs)是与几丁质代谢以及几丁质转化为壳聚糖相关的碳水化合物酯酶。研究表明,几丁质脱乙酰作用对于几丁质的组织化和紧密性至关重要,因此会影响几丁质结构(如围食膜(PM)和表皮)的机械性能和通透性。在本研究中,在中国柞蚕幼虫中鉴定并表征了两个编码CDA蛋白异构体的基因(和)。尽管鉴定出了五个特征基序,但CDA5蛋白仅具有几丁质脱乙酰化催化结构域。时空表达模式分析表明,两种转录本在蜕皮后取食期的中肠前部区域丰度最高,表明它们在几丁质周转和围食膜组装中发挥作用。通过RNA干扰(RNAi)下调和与围食膜中几丁质微纤丝的断裂相关,表明V组CDAs对于几丁质层的生长和组装至关重要。此外,和可能具有不重叠的功能,以不同方式调节围食膜几丁质结构的形态特征。注射针对和转录本的双链RNA(dsRNA)的幼虫比对照组对感染更不具抵抗力。这些结果表明,下调和对围食膜结构具有独立影响,并破坏了围食膜的完整性,从而扰乱了围食膜抵抗经口感染微孢子虫的功能。我们的数据为区分鳞翅目昆虫V组CDAs之间的CDA功能提供了新证据。