Expression patterns and antifungal function study of KaSPI in Mythimna separata.

IF 1.6 3区 农林科学 Q2 ENTOMOLOGY Bulletin of Entomological Research Pub Date : 2023-12-01 Epub Date: 2023-09-21 DOI:10.1017/S000748532300041X
Ya-Ru Chen, Hong-Jia Yang, Jin-Myong Cha, Xin-Xin Zhang, Dong Fan
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Abstract

Kazal-type serine protease inhibitors (KaSPI) play important roles in insect growth, development, digestion, metabolism and immune defence. In this study, based on the transcriptome of Mythimna separata, the cDNA sequence of MsKaSPI with Kazal domain was uploaded to GenBank (MN931651). Spatial and temporal expression analysis showed that MsKaSPI was expressed at different developmental stages and different tissues, and it was induced by 20-hydroxyecdysone in third-instar larvae of M. separata. After 24 h infection by Beauveria bassiana, the expression level of MsKaSPI and the corresponding MsKaSPI content were significantly up-regulated, being 6.42-fold and 1.91-fold to the control group, respectively, while the activities of serine protease, trypsin and chymotrypsin were inhibited. After RNA interference interfered with MsKaSPI for 6 h, the expression decreased by 73.44%, the corresponding content of MsKaSPI protein decreased by 55.66% after 12 h, and the activities of serine protease and trypsin were significantly enhanced. Meanwhile, both the larval and pupal stages of M. separata were prolonged, the weights were reduced and the number of eggs per female decreased by 181. Beauveria bassiana infection also increased the mortality of MsKaSPI-silenced M. separata by 18.96%. These prove MsKaSPI can not only result in slow growth and low fecundity of M. separata by regulating the activity of related protease, but also participate in the resistance to pathogenic fungi by regulating the serine protease inhibitor content and the activities of related serine protease.

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KaSPI在粘虫中的表达模式及抗真菌功能研究。
Kazal型丝氨酸蛋白酶抑制剂(KaSPI)在昆虫生长、发育、消化、代谢和免疫防御中发挥着重要作用。在本研究中,基于Mythimna separata的转录组,将具有Kazal结构域的MsKaSPI的cDNA序列上传到GenBank(MN931651)。时空表达分析表明,MsKaSPI在不同发育阶段和不同组织中均有表达,并由20-羟基蜕皮激素在粘虫三龄幼虫中诱导表达。白僵菌感染24小时后,MsKaSPI的表达水平和相应的MsKaSPI含量显著上调,分别是对照组的6.42倍和1.91倍,而丝氨酸蛋白酶、胰蛋白酶和糜蛋白酶的活性受到抑制。RNA干扰MsKaSPI 6h后,MsKaSPI的表达下降了73.44%,12h后相应的MsKaSPI蛋白含量下降了55.66%,丝氨酸蛋白酶和胰蛋白酶的活性显著增强。同时,粘虫的幼虫和蛹期都延长了,重量减轻了,每只雌性的卵数减少了181个。白僵菌感染也使MsKaSPI沉默的分离分枝杆菌的死亡率增加了18.96%。这证明MsKaSPI不仅可以通过调节相关蛋白酶的活性导致分离分枝杆菌生长缓慢和繁殖力低下,还可以通过调节丝氨酸蛋白酶抑制剂的含量和相关丝氨酸蛋白酶的活性参与对病原真菌的抗性。
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来源期刊
CiteScore
4.00
自引率
0.00%
发文量
160
审稿时长
6-12 weeks
期刊介绍: Established in 1910, the internationally recognised Bulletin of Entomological Research aims to further global knowledge of entomology through the generalisation of research findings rather than providing more entomological exceptions. The Bulletin publishes high quality and original research papers, ''critiques'' and review articles concerning insects or other arthropods of economic importance in agriculture, forestry, stored products, biological control, medicine, animal health and natural resource management. The scope of papers addresses the biology, ecology, behaviour, physiology and systematics of individuals and populations, with a particular emphasis upon the major current and emerging pests of agriculture, horticulture and forestry, and vectors of human and animal diseases. This includes the interactions between species (plants, hosts for parasites, natural enemies and whole communities), novel methodological developments, including molecular biology, in an applied context. The Bulletin does not publish the results of pesticide testing or traditional taxonomic revisions.
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