Functional assessment of cadherin as a shared mechanism for cross/dual resistance to Cry1Ac and Cry2Ab in Helicoverpa zea

Helicoverpa zea is a major target pest of pyramided transgenic crops expressing Cry1,Cry2 and/or Vip3Aa proteins from Bacillus thuringiensis(Bt)in the United States.Laboratory-selected Cry1Ac/Cry2Ab cross resistance and fieldevolved practical dual resistance of H.zea to these two toxins have been widely reported.Whether the widespread Cry1Ac/Cy2Ab dual resistance of H.zea has resulted from the selection of one shared or two independent resistance mechanisms by pyramided Bt crops remains unclear.Cadherin is a well-confirmed receptor of Cry1Ac and a suggested receptor of Cry2Ab in at least three Lepidopteran species.To test whether cadherin may serve as one shared mechanism for the cross and dual resistance of H.zea to Cry1Ac and Cry2Ab,we cloned H.zea cadherin(HzCadherin)cDNA and studied its functional roles in the mode of action of Cry1Ac and Cry2Ab by gain-and lossof-function analyses.Heterologous expression of HzCadherin in H.zea midgut,H.zea fat body and Sf9 cells made all three of these cell lines more susceptible to activated Cry1Ac but not activated Cry2Ab,whereas silencing HzCadherin of H.zea midgut and fat body cells significantly reduced the susceptibility to Cry1Ac but not Cry2Ab.Likewise,suppressing HzCadherin with siRNA made H.zea larvae resistant to Cry1Ac.These results clearly demonstrate that HzCadherin is not a receptor for Cry2Ab,and thus it is unlikely to serve as one shared mechanism for the cross and dual resistance of H.zea to Cry1Ac and Cry2Ab.

Risk Assessment and Ecological Effects of Transgenic Bacillus thuringiensis Crops on Non-Target Organisms

The application of recombinant DNA technology has resulted in many insect-resistant varieties by genetic engineering （GE）. Crops expressing Cry toxins derived from Bacillus thuringiensis （Bt） have been planted worldwide, and are an effective tool for pest control. However, one ecological concern regarding the potential effects of insect-resistant GE plants on non-target organisms （NTOs） has been continually debated. In the present study, we briefly summarize the data regarding the development and commercial use of transgenic Bt varieties, elaborate on the procedure and methods for assessing the non-target effects of insect-resistant GE plants, and synthetically analyze the related research results, mostly those published between 2005 and 2010. A mass of laboratory and field studies have shown that the currently available Bt crops have no direct detrimental effects on NTOs due to their narrow spectrum of activity, and Bt crops are increasing the abundance of some beneficial insects and improving the natural control of specific pests. The use of Bt crops, such as Bt maize and Bt cotton, results in significant reductions of insecticide application and clear benefits on the environment and farmer health. Consequently, Bt crops can be a useful component of integrated pest management systems to protect the crop from targeted pests.

Inductive and synergistic interactions between plant allelochemical flavone and Bt toxin Cry1 Ac in Helicoverpa armigera

Genetically engineered crops simultaneously produce defensive allelochemi-cals and Bacillus thuringiensis(Bt)toxin proteins to kill some of the world's most devastating insect pests.How the two types of toxins,when ingested sequentially or simultaneously,interact at both lethal and sublethal doses in these pests remains underexplored.Here,we examined the toxicological interactions between the Bt toxin Cry 1 Ac and the flavonoid allelochemical flavone in Helicoverpa armigera.Simultaneous exposure of H.armigera neonates to lethal doses(LC25)of Cry 1 Ac and flavone caused a mortality significantly higher than that of either toxin alone and their expected additive mortality.Preexposure for 24 h to a sublethal dose(LC10)of Cry 1 Ac followed by 6-d simultaneous exposure to the same dose of Cry 1 Ac plus a lethal dose(1.6 mg/g diets,LC50)of flavone resulted in a mortality significantly higher than that of the LC50 dose of flavone alone and the expected additive mortality of the LC50 dose of flavone plus the LC10 dose of Cry 1 Ac.One-day preexposure to the sublethal dose(LC10)of flavone followed by 6-d simultaneous exposure to the LC50 dose(6 ng/cm2)of Cry 1 Ac plus the LC25 dose of flavone yielded a mortality significantly higher than that of the LC50 dose of Cry 1 Ac but similar to the expected additive mortality of the LC50 dose of Cry 1 Ac plus the LC50 dose of flavone.The results suggest that Cry 1 Ac induces and synergizes the toxicity of flavone against H.armigera larvae.