Department of Plant Protection, College of Agronomy, Northeast Agricultural University, Harbin, China.
Department of Microbiology, Suifenhe Customs, Suifenhe, China.
Arch Insect Biochem Physiol. 2020 May;104(1):e21666. doi: 10.1002/arch.21666. Epub 2020 Feb 29.
Chitin deacetylase (CDA) is a hydrolytic enzyme that modifies chitin into chitosan in the body of insects. In this study, we obtained a full-length complementary DNA sequence (MsCDA1) from the oriental armyworm Mythimna separata by high-throughput sequencing. MsCDA1 is 1,952 bp long and includes 1,620 bp open reading frame encoding 539 amino acids. Analysis by quantitative real time polymerase chain reaction showed that MsCDA1 expression was higher at the adult stage than at earlier developmental stages. MsCDA1 was expressed in all larval tissues examined, in which the highest expression level was found in the midgut. The RNA interference (RNAi) suppressed MsCDA1 expression levels at 12, 24, and 48 hr after injection of double-stranded RNA (1-4 μg per larva) specific to MsCDA1. Under RNAi condition, CDA enzyme activity was significantly reduced and changes an ultramicroscopic structure of M. separata peritrophic matrix especially in its microfibrillar organization exhibiting loose network. In contrast, the surface of the peritrophic matrix was relatively smooth and well organized at control or low RNAi conditions. Moreover, RNAi of MsCDA1 expression impaired larval growth and development, occasionally leading to larval death. These results demonstrate that MsCDA1 plays a crucial role in maintaining peritrophic matrix integrity in M. separata.
几丁质脱乙酰酶(CDA)是一种水解酶,可在昆虫体内将几丁质转化为壳聚糖。在本研究中,我们通过高通量测序从东方粘虫 My thimna separata 中获得了全长互补 DNA 序列(MsCDA1)。MsCDA1 长 1952bp,包含 1620bp 的开放阅读框,编码 539 个氨基酸。定量实时聚合酶链反应分析表明,MsCDA1 在成虫期的表达高于早期发育阶段。MsCDA1 在所有检测到的幼虫组织中表达,其中在中肠中的表达水平最高。RNA 干扰(RNAi)抑制了特异性针对 MsCDA1 的双链 RNA(每只幼虫 1-4μg)注射后 12、24 和 48 小时的 MsCDA1 表达水平。在 RNAi 条件下,CDA 酶活性显著降低,并且东方粘虫的围食膜的超微结构发生变化,特别是在其微纤维组织中呈现疏松的网络。相比之下,在对照或低 RNAi 条件下,围食膜的表面相对光滑且组织良好。此外,MsCDA1 表达的 RNAi 损害了幼虫的生长和发育,偶尔导致幼虫死亡。这些结果表明 MsCDA1 在维持东方粘虫围食膜完整性中起着至关重要的作用。