[1]陈晓,吴焱锟,李会琴,等.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(07):1349-1359.[doi:doi:10.3969/j.issn.1000-4440.2026.07.006]
　CHEN Xiao,WU Yankun,LI Huiqin,et al.Cloning and expression analysis of serine protease inhibitor genes EfSPI8 and EfSPI31 in Eocanthecona furcellata[J].,2026,42(07):1349-1359.[doi:doi:10.3969/j.issn.1000-4440.2026.07.006]
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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()

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
42
期数:
2026年07期
页码:
1349-1359
栏目:
植物保护
出版日期:
2026-07-31

文章信息/Info

Title:
Cloning and expression analysis of serine protease inhibitor genes EfSPI8 and EfSPI31 in Eocanthecona furcellata
作者:
陈晓1; 吴焱锟1; 李会琴1; 杨昌和1; 陈岗2; 马云昆3; 吴国星1; 高熹1
(1.云南农业大学植物保护学院,云南昆明650201;2.云南省烟草公司楚雄州公司,云南楚雄675000；3.安宁市种植业服务中心,云南昆明650303)
Author(s):
CHEN Xiao1; WU Yankun1; LI Huiqin1; YANG Changhe1; CHEN Gang2; MA Yunkun3; WU Guoxing1; GAO Xi1
(1.College of Plant Protection, Yunnan Agricultural University, Kunming 650201, China;2.Chuxiong Prefecture Company, Yunnan Tobacco Company, Chuxiong 675000, China;3.Anning Planting Industry Service Center, Kunming 650303, China)
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
Keywords:
Eocanthecona furcellata; salivary gland; serine protease inhibitor; gene cloning; expression pattern
分类号:
S476.2
DOI:
doi:10.3969/j.issn.1000-4440.2026.07.006
文献标志码:
A
摘要:
丝氨酸蛋白酶抑制剂(SPI)参与昆虫凝血、免疫黑化及生长发育等关键生理过程的调控,在昆虫免疫及宿主-病原体互作中发挥重要作用。本研究旨在克隆叉角厉蝽毒液丝氨酸蛋白酶抑制剂编码基因EfSPI8和EfSPI31,并分析其编码蛋白质的特性及表达模式。通过PCR成功克隆获得EfSPI8、EfSPI31的开放阅读框(ORF)序列,长度分别为1 083 bp、2 793 bp,分别编码361个、931个氨基酸。生物信息学分析结果表明,EfSPI8、EfSPI31蛋白的理论相对分子量分别为41 100、104 800,两者N端分别含有长度为30 aa、26 aa的信号肽序列,均属于分泌蛋白；EfSPI8具有典型的丝氨酸蛋白酶抑制剂超家族（Serpin）结构域及反应中心环(RCL),属于Serpin超家族；EfSPI31含有3对二硫键以稳定蛋白质结构,属于Kazal型丝氨酸蛋白酶抑制剂家族。序列比对结果显示,EfSPI8与茶翅蝽(Halyomorpha halys)Serpin的同源性为39.27%；EfSPI31与稻绿蝽(Nezara viridula)Kazal的同源性达88.28%。实时荧光定量PCR(RT-qPCR)结果显示,EfSPI8基因在叉角厉蝽5龄若虫、雄成虫、唾液腺主腺前叶及精巢中高表达,推测其可能参与捕食相关免疫抑制及生殖调控；EfSPI31基因在4～5龄叉角厉蝽若虫、精巢和脂肪体中高表达,推测其可能与免疫、代谢储备和生殖系统发育相关。研究结果为阐明2种蛋白酶抑制剂在捕食性天敌昆虫中的多功能作用提供了分子基础。
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 insect immunity and host-pathogen interactions. This study aimed to clone the venom serine protease inhibitor genes EfSPI8 and EfSPI31 from the predatory stink bug Eocanthecona furcellata and analyze the characteristics and expression patterns of their encoded proteins. The open reading frame (ORF) sequences of EfSPI8 and EfSPI31 were successfully cloned by PCR, with lengths of 1 083 bp and 2 793 bp, encoding 361 and 931 amino acids, respectively. Bioinformatic analysis revealed that the theoretical relative molecular masses of EfSPI8 and EfSPI31 were 41 100 and 104 800, respectively. Both proteins contained signal peptide sequences at their N-termini, with lengths of 30 and 26 amino acids, indicating 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.

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备注/Memo

备注/Memo:
收稿日期：2025-11-25基金项目：云南省农业基础研究联合专项(202301BD070001-137)；云南省中青年学术和技术带头人后备人才项目(202205AC160077)；云南省烟草公司科技计划项目(2023530000241012)作者简介：陈晓(2001-),女,云南芒市人,硕士研究生,研究方向为昆虫分子生物学与生物防治。(E-mail)15108646682@163.com通讯作者：高熹,(E-mail)chonchon@163.com
更新日期/Last Update: 2026-08-21