烟草粉螟卵黄原蛋白基因鉴定及其对X射线胁迫的响应  

Identification of vitellogenin gene in Ephestia elutella and its response to X-ray irradiation stress

作　　者：赵钧 徐鹿[2] 苗圃[3] 李成军[1] 吴孔明[4] 李淑君[1] ZHAO Jun;XU Lu;MIAO Pu;LI Chengjun;WU Kongming;LI Shujun(State Tobacco Monopoly Administration Key Laboratory for Green Prevention and Control of Tobacco Diseases and Pests in Huanghuai Growing Area,Institute of Tobacco Research,Henan Academy of Agricultural Sciences,Xuchang 461000,China;Key Lab of Food Quality and Safety of Jiangsu Province-State Key Laboratory Breeding Base,Institute of Plant Protection,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;Luoyang Branch of Henan Tobacco Company,Luoyang 471000,China;Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China)

机构地区：[1]河南省农业科学院烟草研究所,烟草行业黄淮烟区烟草病虫害绿色防控重点实验室,许昌461000 [2]江苏省农业科学院植物保护研究所,省部共建国家重点实验室培育基地-江苏省食品质量安全重点实验室,南京210014 [3]河南省烟草公司洛阳市公司,洛阳471000 [4]中国农业科学院植物保护研究所,北京100193

出　　处：《中国烟草学报》2024年第3期132-138,共7页Acta Tabacaria Sinica

基　　金：中国烟草总公司河南省公司科技项目(No.2021410000240023);河南省农业科学院自主创新项目(No.2023ZC024);国家自然科学基金面上项目(No.31972309)。

摘　　要：【背景和目的】卵黄原蛋白(vitellogenin,Vg)是昆虫卵黄发生的关键物质,为探索X射线辐照胁迫对烟草粉螟Vg基因转录表达和雌成虫繁殖力的影响。【方法】采用RT-PCR技术克隆烟草粉螟Vg基因的全长并进行生物信息学分析,利用qPCR技术检测在烟草粉螟雌成虫不同组织(足、头、中肠、脂肪体、卵巢和表皮)的表达量和雌成虫在不同X射线辐照剂量(50,100和150 Gy)胁迫下该基因的相对表达量以及对其蛹羽化率、成虫寿命、产卵量、卵孵化率的影响。【结果】(1)克隆获得烟草粉螟卵黄原蛋白基因,命名为EeVg(GenBank登录号:OM804251),开放阅读框(ORF)长度为5337bp,编码1778个氨基酸。(2)EeVg基因在雌成虫的脂肪体中相对表达量最高,为头部表达量的17.96倍。(3)100和150 Gy的X射线辐照雌蛹极显著地降低EeVg在烟草粉螟羽化后的表达量,分别为对照的0.34倍和0.09倍。(4)剂量为50、100和150 Gy的X射线辐照降低烟草粉螟雌蛹羽化率和成虫寿命(87.78%~75.56%和6.40~4.17d),显著降低雌蛾的产卵量和卵孵化率(55.27~20.37粒和76.67%~72.22%)。【结论】高剂量的X射线辐照胁迫可使EeVg基因表达量显著下调,显著降低烟草粉螟繁殖力,为深入研究X射线影响烟草粉螟繁殖力的分子机制奠定了基础。[Background]Vitellogenin(Vg)is the precursor protein of egg yolk vitellin(Vn)in insects.This study aims to explore the effects of X-ray irradiation stress on the transcript expression of Vg gene and fecundity of tobacco moth Ephestia elutella,in order to lay the foundation for revealing the molecular mechanism.[Methods]The full length of the EeVg gene was cloned by RT-PCR and analyzed by bioinformatics methods.RT-qPCR technology was used to detect the expression patterns of the gene in different tissues(foot,head,midgut,fat body,ovary and epidermis).Female pupae of E.elutella were irradiated via an X-ray irradiator X-RAD 320.Then the effects of X-ray irradiation at the doses of 50,100 and 150 Gy on the emergence rate of female pupae,adult longevity,egg number,egg hatch rate were observed,and the relative expression of EeVg gene were measured by qPCR.[Results]1)The vitellogenin protein gene,namely EeVg(GenBank No.OM804251),was cloned and identified from E.elutella.Its ORF length was 5337 bp,encoding 1,778 amino acids.2)The relative expression level of the EeVg gene was highest in the fat body of the female adult,which was 17.96 times that of the head.3)X-ray irradiation at 100 and 150 Gy significantly down-regulated the relative expression of EeVg in female pupae after emergence by 0.34 and 0.09 times,respectively.4)X-ray irradiation at doses of 50,100,and 150 Gy reduced the pupation rate and adult lifespan of the tobacco powder moth(87.78%~75.56%and 6.40~4.17 d),significantly reducing the oviposition quantity and egg hatching rate of female moths(55.27~20.37 eggs and 76.67%~72.22%).[Conclusion]X-ray irradiation stress down-regulated EeVg gene expression and significantly reduced the fecundity of female adults,which lays a foundation for further study of the molecular mechanism of X-ray affecting the fecundity of E.elutella.

关 键 词：烟草粉螟 卵黄原蛋白基因 X射线辐照 表达量 繁殖力 

分 类 号：S435.72[农业科学—农业昆虫与害虫防治]