棉铃虫中肠钙粘蛋白、氨肽酶N及碱性磷酸酯酶的抗血清对Cry1Ac和Cry2Aa毒力的影响

doi:10.16380/j.kcxb.2016.09.007

摘要/Abstract

摘要： 【目的】Cry1A和Cry2A 类Bt蛋白通过特异性地与昆虫中肠上的受体蛋白结合而发挥杀虫作用，现已广泛应用于转基因抗虫作物。本研究旨在进一步明确Cry2A 类蛋白的作用机制和Cry1A受体蛋白在Cry2A发挥毒力中的作用。【方法】本研究首先提取了棉铃虫 Helicoverpa armigera的BBMV，制备了钙粘蛋白（CAD）、氨肽酶N（APN）和碱性磷酸酯酶（ALP）3种受体蛋白的抗体和抗血清；然后，利用Western blot检测BBMV上这3种受体蛋白后，利用抗体封闭技术比较了敏感棉铃虫和Cry1Ac抗性棉铃虫（BtR）中3种受体蛋白的抗血清对Cry1Ac和Cry2Aa毒力的影响。【结果】对敏感品系棉铃虫，这3种已知的Cry1Ac受体蛋白抗血清显著地降低了Cry1Ac和Cry2Aa的毒力。其中APN抗血清对Cry1Ac毒力的影响最大，棉铃虫幼虫的死亡率降低了84.44%；ALP抗血清对Cry2Aa的毒力影响最大，棉铃虫幼虫死亡率比对照降低了71.04%。Cry1Ac对Cry1Ac抗性棉铃虫（BtR）的毒力显著降低，Cry2Aa的毒性也减弱。在Cry1Ac抗性棉铃虫（BtR）中，3种受体抗血清对Cry1Ac的影响比在敏感棉铃虫中的影响小，尤其是CAD和APN抗血清对Cry1Ac毒力的抑制率显著低于在敏感棉铃虫中的抑制作用；CAD和ALP抗血清对Cry2Aa毒力的影响与在敏感棉铃虫中的影响差异不显著，但APN抗血清可以显著降低Cry2Aa对Cry1Ac抗性棉铃虫（BtR）的毒力。【结论】棉铃虫CAD，APN和ALP不仅参与了Cry1Ac的杀虫过程，也对Cry2Aa毒力有一定的影响，而且这3种蛋白可能与棉铃虫对Cry1Ac和Cry2Aa产生抗性及交互抗性相关。

关键词: 棉铃虫, 受体蛋白, 抗血清, Cry1Ac, Cry2Aa, 抗性, 交互抗性

Abstract: 【Aim】 Cry1A and Cry2A toxins play insecticidal roles by specifically binding with receptor proteins on insect midgut, and are widely used in Bt transgenic crops now. This study aims to further understand the action mechanisms of Cry2A and the roles of the functional receptor proteins of Cry1A in the toxicity of Cry2A. 【Methods】 We firstly extracted brush border membrane vesicles (BBMV) of midgut, prepared antibody and antiserums of cadherin (CAD), aminopeptidase N (APN), and alkaline phosphatase (ALP) in Helicoverpa armigera. Then, after detecting the antiserums of these three receptor proteins in BBMV by Western blot, we compared the effects of antiserums of these three receptor proteins on the toxicities of Cry1Ac and Cry2Aa in the susceptible and Cry1Ac-resistant H. armigera (BtR) by the antibody blocking technology. 【Results】 For the susceptible H. armigera, the antiserums of these three receptor proteins not only significantly reduced the toxicity of Cry1Ac, but also remarkably reduced the toxicity of Cry2Aa. Among them, anti-APN antiserum had the biggest impact on the toxicity of Cry1Ac, causing the mortality of H. armigera larvae to be reduced by 84.44%, and anti-ALP antiserum had the greatest effect on the toxicity of Cry2Aa, causing the larval mortality to be reduced by 71.04% compared with the control (without treatment by antiserum). The toxicity of Cry1Ac to Cry1Ac-resistant H. armigera (BtR) was obviously reduced and the toxicity of Cry2Aa also became less to these resistant larvae. The influences of the antiserums of these three receptors on the toxicity of Cry1Ac to the Cry1Ac-resistant strain of H. armigera (BtR) were smaller than those to the susceptible strain. Especially, the inhibition percentage of anti-CAD and -APN antiserums to the toxicity of Cry1Ac decreased significantly. The effects of anti-CAD and -ALP antiserums on the toxicity of Cry2Aa to the Cry1Ac-resistant strain (BtR) and the susceptible strain were not significantly different, but the anti-APN antiserum obviously reduced the toxicity of Cry2Aa to the Cry1Ac-resistant strain (BtR). 【Conclusion】 The midgut receptor proteins CAD, APN and ALP not only participate in the insecticidal process of Cry1Ac, but also affect the toxicity of Cry2Aa to H. armigera, and these three receptors may be related with the resistance and cross-resistance to Cry1Ac and Cry2Aa in H. armigera.

Key words: Helicoverpa armigera, receptor protein, antiserum, Cry1Ac, Cry2Aa, resistance, cross-resistance

王冰洁, 袁向东, 赵曼, 刘臣, 陈琳, 梁革梅. 棉铃虫中肠钙粘蛋白、氨肽酶N及碱性磷酸酯酶的抗血清对Cry1Ac和Cry2Aa毒力的影响[J]. 昆虫学报, 2016, 59(9): 977-984.

WANG Bing-Jie, YUAN Xiang-Dong, ZHAO Man, LIU Chen, CHEN Lin, LIANG Ge-Mei. Effects of antiserums of cadherin, aminopeptidase N and alkaline phosphatase on the toxicities of Cry1Ac and Cry2Aa in Helicoverpa armigera (Lepidoptera: Noctuidae)[J]. Acta Entomologica Sinica, 2016, 59(9): 977-984.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: http://www.insect.org.cn/CN/10.16380/j.kcxb.2016.09.007

http://www.insect.org.cn/CN/Y2016/V59/I9/977

[1]	王婷婷, 索倩, 孙晓燕, 马跃敏, 刘凯于, 彭建新, 彭蓉 . 棉铃虫激活增强子结合蛋白AP-4基因的原核表达、多克隆抗体制备、表达谱分析及功能鉴定[J]. 昆虫学报, 2019, 62(9): 1028-1037.
[2]	王芹芹, 崔丽, 王奇渊, 王立, 谢娜, 袁会珠, 芮昌辉. 茚虫威亚致死浓度对茚虫威敏感性降低的棉铃虫生物学参数及解毒酶活性的影响[J]. 昆虫学报, 2019, 62(8): 960-969.
[3]	张亚坤, 邓中原, 谷少华, 李显春. 棉铃虫剂量补偿相关基因Hamsl1的鉴定与功能分析[J]. 昆虫学报, 2019, 62(7): 799-813.
[4]	赵洁, 魏倩, 任苏伟, 刘小宁. 棉铃虫叉头框蛋白A类似蛋白基因HarmFoxAl的克隆及表达谱分析[J]. 昆虫学报, 2019, 62(6): 672-684 .
[5]	刘小飞, 孟建玉, 赵晓超, 张长禹. 棉铃虫c-Jun氨基末端激酶基因的克隆、表达谱及对UV-A胁迫的响应[J]. 昆虫学报, 2019, 62(4): 407-417.
[6]	姚雪, 李琳红, 魏纪珍, 席玉强, 杜孟芳, 崔江宽, 安世恒, 刘晓光. 棉铃虫酪氨酸羟化酶基因的分子特性及功能分析[J]. 昆虫学报, 2019, 62(3): 294-303.
[7]	麦麦提艾力·阿卜杜纳斯尔, 马纪, 刘宁, 王威, 李梦鸽, 古新蓉, 王新, 刘小宁. 棉铃虫Ⅳ型几丁质酶的基因克隆、酶学性质及表达谱[J]. 昆虫学报, 2019, 62(2): 170-180.
[8]	梁振普, 王亮, 李鹏娟, 王朝兴, 肖宇博, 张小霞. 棉铃虫围食膜的蛋白质组鉴定[J]. 昆虫学报, 2019, 62(2): 181-192.
[9]	艾东, 林荣华, 王猛, 梁晓贺, 于彩虹. 棉铃虫可溶型海藻糖酶的化学修饰[J]. , 2018, 61(7): 801-807.
[10]	闫硕, 刘彦君, 张馨方, 朱家林, 李贞, 刘孝明, 张青文, 刘小侠. 棉铃虫生物钟基因HeDbt的克隆和表达模式分析[J]. 昆虫学报, 2018, 61(12): 1393-1403.
[11]	张夏瑄, 郭孟博, 刘一鹏, 王冰, 王桂荣. 棉铃虫离子型受体基因HarmIR8a的克隆及表达定位[J]. 昆虫学报, 2018, 61(11): 1263-1271.
[12]	高景鹏, 陈翰明, 马雅军. 我国白纹伊蚊对三种拟除虫菊酯类杀虫剂抗性检测诊断剂量的建立[J]. , 2018, 61(1): 18-24.
[13]	张甜甜, 苗娅, 许柏英, 陈斌. 中华按蚊α-羧酸酯酶AsAe7的异源表达、纯化与结晶[J]. , 2018, 61(1): 48-58.
[14]	王芮, 张浩淼, 邓蕾, 肖倩倩, 王秋霞, 刘广娜, 韩召军, 吴敏. 灰飞虱羧酸酯酶LsCarE1介导杀虫剂抗性[J]. , 2017, 60(9): 1006-1012.
[15]	李兆群, 张帅, 周淑芬, 雒珺瑜, 崔金杰. 棉铃虫气味结合蛋白HarmOBP16的组织表达谱及配基结合特性分析[J]. , 2017, 60(8): 891-899.

棉铃虫中肠钙粘蛋白、氨肽酶N及碱性磷酸酯酶的抗血清对Cry1Ac和Cry2Aa毒力的影响

Effects of antiserums of cadherin, aminopeptidase N and alkaline phosphatase on the toxicities of Cry1Ac and Cry2Aa in Helicoverpa armigera (Lepidoptera: Noctuidae)

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 10