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Chitin deacetylase: A potential target for Mythimna separata (Walker) control.
Archives of Insect Biochemistry and Physiology ( IF 1.5 ) Pub Date : 2020-02-29 , DOI: 10.1002/arch.21666 Yan Liu 1 , Jing Yang 1 , Lei Yao 2 , Shuangyu Li 1 , Yaru Chen 1 , Hongjia Yang 1 , Dong Fan 1
Archives of Insect Biochemistry and Physiology ( IF 1.5 ) Pub Date : 2020-02-29 , DOI: 10.1002/arch.21666 Yan Liu 1 , Jing Yang 1 , Lei Yao 2 , Shuangyu Li 1 , Yaru Chen 1 , Hongjia Yang 1 , Dong Fan 1
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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.
中文翻译：
几丁质脱乙酰基酶：分离Mythimna separata（Walker）的潜在目标。
几丁质脱乙酰基酶（CDA）是一种水解酶，可将几丁质变成昆虫体内的壳聚糖。在这项研究中，我们通过高通量测序从东方粘虫Mythimna separata获得了全长互补DNA序列（MsCDA1）。MsCDA1长1,952 bp，包括1,620 bp的开放阅读框，编码539个氨基酸。通过定量实时聚合酶链反应的分析表明，MsCDA1在成人阶段的表达高于早期发育阶段。MsCDA1在所有幼虫组织中均有表达，其中最高表达水平出现在中肠。RNA干扰（RNAi）抑制MsCDA1注射针对MsCDA1的双链RNA（每个幼虫1-4μg）后第12、24和48小时的表达水平。在RNAi条件下，CDA酶的活性显着降低，并改变了分离的分枝杆菌周围营养基质的超微结构，特别是其微纤维组织呈现出松散的网络。相反，在对照或低RNAi条件下，营养周质基质的表面相对光滑且组织良好。此外，MsCDA1表达的RNAi损害幼虫的生长和发育，偶尔导致幼虫死亡。这些结果表明，MsCDA1起着维持围食基质完整性至关重要的作用粘虫。
更新日期：2020-02-29
中文翻译：
几丁质脱乙酰基酶：分离Mythimna separata（Walker）的潜在目标。
几丁质脱乙酰基酶（CDA）是一种水解酶，可将几丁质变成昆虫体内的壳聚糖。在这项研究中，我们通过高通量测序从东方粘虫Mythimna separata获得了全长互补DNA序列（MsCDA1）。MsCDA1长1,952 bp，包括1,620 bp的开放阅读框，编码539个氨基酸。通过定量实时聚合酶链反应的分析表明，MsCDA1在成人阶段的表达高于早期发育阶段。MsCDA1在所有幼虫组织中均有表达，其中最高表达水平出现在中肠。RNA干扰（RNAi）抑制MsCDA1注射针对MsCDA1的双链RNA（每个幼虫1-4μg）后第12、24和48小时的表达水平。在RNAi条件下，CDA酶的活性显着降低，并改变了分离的分枝杆菌周围营养基质的超微结构，特别是其微纤维组织呈现出松散的网络。相反，在对照或低RNAi条件下，营养周质基质的表面相对光滑且组织良好。此外，MsCDA1表达的RNAi损害幼虫的生长和发育，偶尔导致幼虫死亡。这些结果表明，MsCDA1起着维持围食基质完整性至关重要的作用粘虫。
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