Akhurst Raymond Joseph, James William, Bird Lisa Jane, Beard Cheryl
Commonwealth Scientific and Industrial Research Organisation, Division of Entomology, Canberra ACT 2601, Australia.
J Econ Entomol. 2003 Aug;96(4):1290-9. doi: 10.1603/0022-0493-96.4.1290.
Three laboratory strains of Helicoverpa armigera (Hübner) were established by mating of field-collected insects with an existing insecticide-susceptible laboratory strain. These strains were cultured on artificial diet containing the Cry1Ac protoxin of Bacillus thuringiensis using three different protocols. When no response to selection was detected after 7-11 generations of selection, the three strains were combined by controlled mating to preserve genetic diversity. The composite strain (BX) was selected on the basis of growth rate on artificial diet containing Cry1Ac crystals. Resistance to Cry1Ac was first detected after 16 generations of continuous selection. The resistance ratio (RR) peaked approximately 300-fold at generation 21, after which it declined to oscillate between 57- and 111-fold. First-instar H. armigera from generation 25 (RR = 63) were able to complete their larval development on transgenic cotton expressing Cry1Ac and produce fertile adults. There appeared to be a fitness cost associated with resistance on cotton and on artificial diet. The BX strain was not resistant to the commercial Bt spray formulations DiPel and XenTari, which contain multiple insecticidal crystal proteins, but was resistant to the MVP formulation, which only contains Cry1Ac. The strain was also resistant to Cry1Ab but not to Cry2Aa or Cry2Ab. Toxin binding assays showed that the resistant insects lacked the high affinity binding site that was detected in early generations of the strain. Genetic analysis confirmed that resistance in the BX strain of H. armigera is incompletely recessive.
通过将野外采集的昆虫与现有的对杀虫剂敏感的实验室品系进行交配,建立了三种棉铃虫(Helicoverpa armigera (Hübner))实验室品系。这些品系使用三种不同的方案,在含有苏云金芽孢杆菌Cry1Ac原毒素的人工饲料上进行培养。在经过7至11代的选择后未检测到对选择的反应时,通过控制交配将这三个品系合并以保持遗传多样性。基于在含有Cry1Ac晶体的人工饲料上的生长速率,选择了复合品系(BX)。在连续选择16代后首次检测到对Cry1Ac的抗性。抗性比(RR)在第21代时达到约300倍的峰值,之后下降至在57倍至111倍之间波动。来自第25代(RR = 63)的一龄棉铃虫能够在表达Cry1Ac的转基因棉花上完成其幼虫发育并产生可育成虫。在棉花和人工饲料上,抗性似乎伴随着适合度代价。BX品系对含有多种杀虫晶体蛋白的商业Bt喷雾制剂DiPel和XenTari不具有抗性,但对仅含有Cry1Ac的MVP制剂具有抗性。该品系对Cry1Ab也具有抗性,但对Cry2Aa或Cry2Ab不具有抗性。毒素结合试验表明,抗性昆虫缺乏在该品系早期世代中检测到的高亲和力结合位点。遗传分析证实,棉铃虫BX品系中的抗性是不完全隐性的。