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对苏云金芽孢杆菌玉米韧皮部汁液以及非靶标植食性昆虫禾谷缢管蚜(同翅目:蚜科)进行免疫分析,检测其中Cry1Ab的存在情况。

Immunological analysis of phloem sap of Bacillus thuringiensis corn and of the nontarget herbivore Rhopalosiphum padi (Homoptera: Aphididae) for the presence of Cry1Ab.

作者信息

Raps A, Kehr J, Gugerli P, Moar W J, Bigler F, Hilbeck A

机构信息

Swiss Federal Research Station of Agroecology and Agriculture, CH-8046 Zürich, Switzerland.

出版信息

Mol Ecol. 2001 Feb;10(2):525-33. doi: 10.1046/j.1365-294x.2001.01236.x.

Abstract

Phloem sap of transgenic Bacillus thuringiensis (Bt) corn expressing a truncated form of the B. thuringiensis delta-endotoxin Cry1Ab, sap sucking aphids feeding on Bt corn and their honeydew were analysed for presence of Cry1Ab using ELISA. Phloem sap of Bt and non-Bt corn was collected using a newly developed technique with a microcapillary being directly inserted into the phloem tubes. Using this technique, no Cry1Ab was detected in the phloem sap. In contrast, measurable concentrations of Cry1Ab in the range of 1 ppb were detected when phloem sap of pooled leaf samples was extracted using EDTA buffer. This was probably because of Cry1Ab toxin released from damaged cells. When analysing apterous adults of Rhopalosiphum padi L. and their honeydew, no Cry1Ab could be detected. In contrast, Cry1Ab was clearly detected in both larvae of the leaf chewing herbivore Spodoptera littoralis (Boisduval) and their faeces, showing that Cry1Ab is detectable after ingestion and excretion by herbivores. These results suggest that R. padi ingests or contains no or only very low concentrations of Cry1Ab in the range of the detection limit. In consequence it is hypothesized that R. padi as an important prey for beneficial insects in corn is unlikely to cause any harm to its antagonists due to mediating Bt toxin.

摘要

采用酶联免疫吸附测定法(ELISA),对表达苏云金芽孢杆菌(Bt)截短型δ-内毒素Cry1Ab的转基因Bt玉米的韧皮部汁液、取食Bt玉米的刺吸式蚜虫及其蜜露进行分析,检测Cry1Ab的存在情况。使用一种新开发的技术,将微量毛细管直接插入韧皮部导管,收集Bt玉米和非Bt玉米的韧皮部汁液。采用该技术,在韧皮部汁液中未检测到Cry1Ab。相比之下,当使用乙二胺四乙酸(EDTA)缓冲液提取混合叶片样本的韧皮部汁液时,检测到了浓度在1 ppb范围内的可测量Cry1Ab。这可能是由于受损细胞释放出Cry1Ab毒素。在分析禾谷缢管蚜无翅成虫及其蜜露时,未检测到Cry1Ab。相比之下,在咀嚼式食叶害虫草地贪夜蛾幼虫及其粪便中均清晰检测到Cry1Ab,表明Cry1Ab在被食草动物摄入和排泄后仍可被检测到。这些结果表明,禾谷缢管蚜摄入或含有的Cry1Ab浓度在检测限范围内不存在或仅非常低。因此,据推测,作为玉米中有益昆虫重要猎物的禾谷缢管蚜,不太可能因介导Bt毒素而对其天敌造成任何危害。

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