Rodrigo-Simón Ana, de Maagd Ruud A, Avilla Carlos, Bakker Petra L, Molthoff Jos, González-Zamora Jose E, Ferré Juan
Department of Genetics, University of Valencia, 46100-Burjassot (Valencia), Spain.
Appl Environ Microbiol. 2006 Feb;72(2):1595-603. doi: 10.1128/AEM.72.2.1595-1603.2006.
The effect of Cry proteins of Bacillus thuringiensis on the green lacewing (Chrysoperla carnea) was studied by using a holistic approach which consisted of independent, complementary experimental strategies. Tritrophic experiments were performed, in which lacewing larvae were fed Helicoverpa armigera larvae reared on Cry1Ac, Cry1Ab, or Cry2Ab toxins. In complementary experiments, a predetermined amount of purified Cry1Ac was directly fed to lacewing larvae. In both experiments no effects on prey utilization or fitness parameters were found. Since binding to the midgut is an indispensable step for toxicity of Cry proteins to known target insects, we hypothesized that specific binding of the Cry1A proteins should be found if the proteins were toxic to the green lacewing. In control experiments, Cry1Ac was detected bound to the midgut epithelium of intoxicated H. armigera larvae, and cell damage was observed. However, no binding or histopathological effects of the toxin were found in tissue sections of lacewing larvae. Similarly, Cry1Ab or Cry1Ac bound in a specific manner to brush border membrane vesicles from Spodoptera exigua but not to similar fractions from green lacewing larvae. The in vivo and in vitro binding results strongly suggest that the lacewing larval midgut lacks specific receptors for Cry1Ab or Cry1Ac. These results agree with those obtained in bioassays, and we concluded that the Cry toxins tested, even at concentrations higher than those expected in real-life situations, do not have a detrimental effect on the green lacewing when they are ingested either directly or through the prey.
通过采用由独立且互补的实验策略组成的整体方法,研究了苏云金芽孢杆菌的Cry蛋白对草蛉(Chrysoperla carnea)的影响。进行了三级营养实验,其中给草蛉幼虫喂食以Cry1Ac、Cry1Ab或Cry2Ab毒素饲养的棉铃虫幼虫。在补充实验中,将预定量的纯化Cry1Ac直接喂给草蛉幼虫。在这两个实验中,均未发现对猎物利用或适合度参数有影响。由于与中肠结合是Cry蛋白对已知靶标昆虫产生毒性必不可少的步骤,我们推测,如果这些蛋白对草蛉有毒,应该能发现Cry1A蛋白的特异性结合。在对照实验中,检测到Cry1Ac与中毒棉铃虫幼虫的中肠上皮结合,并观察到细胞损伤。然而,在草蛉幼虫的组织切片中未发现毒素的结合或组织病理学效应。同样,Cry1Ab或Cry1Ac以特定方式与甜菜夜蛾的刷状缘膜囊泡结合,但不与草蛉幼虫的类似部分结合。体内和体外结合结果强烈表明,草蛉幼虫中肠缺乏Cry1Ab或Cry1Ac的特异性受体。这些结果与生物测定中获得的结果一致,我们得出结论,所测试的Cry毒素,即使浓度高于实际情况中预期的浓度,当它们直接摄入或通过猎物摄入时,对草蛉也没有有害影响。