[1]刘媛,林洁茹,林曼曼,等.Cry2Aa毒素对小菜蛾的杀虫机理[J].江苏农业学报,2018,(04):762-768.[doi:doi:10.3969/j.issn.1000-4440.2018.04.007]
　LIU Yuan,LIN Jie-ru,LIN Man-man,et al.Insecticidal mechanism of Cry2Aa toxin against Plutella xylostella[J].,2018,(04):762-768.[doi:doi:10.3969/j.issn.1000-4440.2018.04.007]
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Cry2Aa毒素对小菜蛾的杀虫机理()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2018年04期
页码:
762-768
栏目:
植物保护
出版日期:
2018-08-25

文章信息/Info

Title:
Insecticidal mechanism of Cry2Aa toxin against Plutella xylostella
作者:
刘媛1; 林洁茹1; 林曼曼1; 张霄1; 徐重新1; 陈蔚1; 罗楚平2; 刘贤金1
（1.江苏省食品质量安全重点实验室，江苏南京210014；2.淮阴工学院，江苏淮安223003）
Author(s):
LIU Yuan1; LIN Jie-ru1; LIN Man-man1; ZHANG Xiao1; XU Chong-xin1; CHEN Wei1; LUO Chu-ping2; LIU Xian-jin1
(1.Jiangsu Key Laboratory for Food Quality and Safety, Nanjing 210014, China；2. Huaiyin Institute of Technology, Huai′an 223003, China)
关键词:
Cry2Aa; 小菜蛾; 杀虫活性; 刷状缘膜囊泡（BBMV）; 免疫组化分析
Keywords:
Cry2Aa; Plutella xylostella; insecticidal activity; brush border membrane vesicle(BBMV); immunohistochemical analysis
分类号:
S482.3+9
DOI:
doi:10.3969/j.issn.1000-4440.2018.04.007
文献标志码:
A
摘要:
为研究苏云金杆菌Cry2Aa毒素的生物活性及杀虫机理，先利用浸叶法测定了商品化的Cry2Aa毒素对小菜蛾的杀虫活性，并用石蜡包埋切片后进行免疫组化分析，再用ELISA法测定了Cry2Aa毒素与小菜蛾刷状缘膜囊泡（BBMV）的结合曲线，最后通过Ligand blot法和肽指纹质谱对小菜蛾BBMV与Cry2Aa毒素的结合蛋白进行了分离与鉴定。结果表明，Cry2Aa毒素对小菜蛾的半数致死浓度（LC50）为27.90 μg/ml。Cry2Aa毒素与小菜蛾中肠上皮细胞存在结合，与小菜蛾BBMV的表观结合亲和力为266.6 nmol/L。小菜蛾BBMV与Cry2Aa毒素存在5条主要结合条带，其中分子量 2.45×105上方的主结合条带酶解后得到ATYSEGPNGSVR片段，通过数据库比对分析，匹配到一个分子量为 3.32×105的小菜蛾未鉴定蛋白，其功能注释为脂质运载蛋白。
Abstract:
To study biological activity and insecticidal mechanism of Bacillus thuringiensis（Bt）Cry2Aa toxin, the insecticidal activity of commercial Cry2Aa toxin against Plutella xylostella was determined by the leaf dipping method, and its paraffin embedded sections were used to perform immunohistochemical analysis. In addition, the binding curve of Cry2Aa toxin with brush border membrane vesicle (BBMV) of P. xylostella was determined by ELISA method. Finally, the binding proteins in BBMV with Cry2Aa were separated and identified by ligand blot and peptide mass fingerprinting. The results showed the half lethal concentration (LC50) of Cry2Aa toxin against P. xylostella was 27.90 μg/ml. Cry2Aa toxin bound to the midgut epithelial cells and the apparent binding affinity of Cry2Aa to BBMV was 266.6 nmol/L. There were five Cry2Aa-binding bands in P. xylostella BBMV. The band on the top of 2.45×105 were digested. A peptide ATYSEGPNGSVR from the band matched to a 3.32×105 uncharactized P. xylostella protein from the database, the functional annotation for the protein was lipocalin.

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备注/Memo

备注/Memo:
收稿日期：2017-12-18 基金项目：国家自然科学基金重点项目（31630061）；江苏省社会发展项目（BE2017706）；江苏省农业自主创新基金项目[CX（17）3018] 作者简介：刘媛（1980-），女，安徽合肥人，硕士，副研究员，主要从事农产品污染物生物控制技术研究。（Tel）025-84390401；（E-mail）zeranol@163.com 通讯作者：刘贤金, （Tel）025-84390401；（E-mail）jaasliu@jaas.ac.cn
更新日期/Last Update: 2018-09-04