江苏农业学报2026，Vol.42Issue(7)：1349-1359,11.DOI:10.3969/j.issn.1000-4440.2026.07.006
叉角厉蝽丝氨酸蛋白酶抑制剂基因EfSPI8、EfSPI31的克隆与表达分析
Cloning and expression analysis of serine protease inhibitor genes EfSPI8 and EfSPI31 in Eocanthecona furcellata
摘要
Abstract
Serine protease inhibitors(SPIs)are involved in regulating key physiological processes in insects,such as coagulation,immune melanization,and growth and development,playing an important role in in-sect immunity and host-pathogen interactions.This study aimed to clone the venom serine protease inhibitor genes EfSPI8 and EfSPI31 from the predatory stink bug Eocan-thecona furcellata and analyze the characteristics and ex-pression patterns of their encoded proteins.The open reading frame(ORF)sequences of EfSPI8 and EfSPI31 were success-fully cloned by PCR,with lengths of 1 083 bp and 2 793 bp,encoding 361 and 931 amino acids,respectively.Bioinformat-ic analysis revealed that the theoretical relative molecular masses of EfSPI8 and EfSPI31 were 41 100 and 104 800,respec-tively.Both proteins contained signal peptide sequences at their N-termini,with lengths of 30 and 26 amino acids,indica-ting that they were secretory proteins.EfSPI8 possessed a typical serine protease inhibitor superfamily(Serpin)domain and a reactive center loop(RCL),belonging to the Serpin superfamily.EfSPI31 contained three pairs of disulfide bonds that stabilized its protein structure,and belonged to the Kazal-type serine protease inhibitor family.Sequence alignment revealed that EfSPI8 shared 39.27%amino acid sequence identity with a Serpin from Halyomorpha halys,while EfSPI31 showed 88.28%identity with a Kazal-type protein from Nezara viridula.Real-time quantitative PCR(RT-qPCR)results showed that EfSPI8 was highly expressed in fifth-instar nymphs,male adults,the anterior lobe of the main salivary gland,and testes,suggesting its potential involvement in prey-related immune suppression and reproductive regulation.EfSPI31 was highly expressed in fourth-to fifth-instar nymphs,testes,and fat body,indicating possible roles in immunity,metabolic reserve,and reproductive system development.These findings provide a molecular basis for understanding the multifunctional roles of these two protease inhibitors in predatory natural enemy insects.关键词
叉角厉蝽/唾液腺/丝氨酸蛋白酶抑制剂/基因克隆/表达模式Key words
Eocanthecona furcellata/salivary gland/serine protease inhibitor/gene cloning/expression pattern分类
农业科技引用本文复制引用
陈晓,吴焱锟,李会琴,杨昌和,陈岗,马云昆,吴国星,高熹..叉角厉蝽丝氨酸蛋白酶抑制剂基因EfSPI8、EfSPI31的克隆与表达分析[J].江苏农业学报,2026,42(7):1349-1359,11.基金项目
云南省农业基础研究联合专项(202301BD070001-137) （202301BD070001-137）
云南省中青年学术和技术带头人后备人才项目(202205AC160077) （202205AC160077）
云南省烟草公司科技计划项目(2023530000241012) （2023530000241012）
