江西省部分地区水稻二化螟抗药性测定

用点滴法普查了江西省南昌、南城、永新、丰城、上饶、彭泽6县(市)二化螟种群4铃幼虫对4种(类)杀虫剂的抗药性。结果表明,供试二化螟种群对沙蚕毒素类农药杀虫单、有机磷类农药三唑磷已产生了较强的抗性,抗性倍数分别为33.30~241.95、7.97~111.20;对苯基吡唑类农药氟虫腈和抗生素类农药阿维茵素部分地区也产生了低水平抗性,抗性倍数分别为5.88~16.47、3.70~9.90。本文还对二化螟抗性治理对策进行了讨论。     

2002年江苏省二化螟抗药性检测及治理

20 0 2年应用点滴法普查了江苏常熟、锡山、武进、句容、姜堰、高邮、楚州、宿豫、灌云及连云港 10县 (市 ) 12个二化螟种群 4龄幼虫对 4种 (类 )杀虫剂的抗药性。结果表明 ,供试种群对沙蚕毒素类农药杀虫单已普遍产生抗性 ,且苏南种群的抗性水平高于苏北 ,常熟为最高 ( 16倍 ) ;对有机磷类农药三唑磷苏南地区种群为低水平抗性 ,其他地区均属敏感水平。目前二化螟供试种群对苯基吡唑类农药氟虫腈和抗生素类农药阿维菌素未产生抗药性。本文还对二化螟抗性治理对策进行了讨论。     

水稻二化螟抗药性监测及防控对策*

采用点滴法测定了2007-2009年采自浙江、江苏、湖北、四川等地水稻二化螟4龄幼虫对沙蚕毒素类的杀虫单、有机磷类的三唑磷、毒死蜱、苯基吡唑类的氟虫腈及微生物源的阿维菌素等杀虫剂的抗性。结果表明：上述二化螟种群对５种杀虫剂抗性分布具有明显的地区性,其中浙江、江苏大部分地区供试种群对杀虫单、三唑磷的抗性水平达40倍以上,已普遍产生高至极高水平抗性;对毒死蜱产生的抗性水平达22倍以上,已普遍产生中等水平至高水平抗性;对氟虫腈产生的抗性水平达6倍以上,已普遍产生低水平至中等水平抗性;对阿维菌素敏感至低水平抗性;湖北孝感、四川武胜种群对５种药剂尚未产生抗性或为低水平抗性。本文还对二化螟抗性防控对策进行了讨论。     

转cry1Ab+cry1C双价抗虫水稻对二化螟的抗性评价

为明确转cry1Ab+cry1C双价抗虫水稻对二化螟的抗性及其杀虫蛋白的时空表达,本研究采用离体水稻组织生测法系统评价了cry1Ab、cry1C、正交cry1Ab+cry1C和反交cry1C+cry1Ab 4种抗虫水稻品系对二化螟的杀虫效果,并用ELISA方法测定了Cry1Ab、Cry1C蛋白在各个抗虫水稻品系中的表达情况。结果显示,二化螟在不同生育期的4种转基因抗虫水稻上取食的叶面积显著低于非转基因对照亲本明恢63。4个转基因抗虫水稻品系在生长前期（苗期、分蘖和拔节期）对二化螟表现极高的杀虫效果,生长后期（孕穗期和成熟期）杀虫效果有所下降。两双价抗虫水稻杀虫效果最好,其次为转cry1Ab和cry1C水稻。Bt蛋白表达水平随着抗虫水稻生育期的变化而变化,且差异显著;Cry1Ab的蛋白表达量在水稻整个生长期均显著高于Cry1C。Cry1Ab蛋白在单价抗虫水稻叶片和茎杆中均表现出随生育期先升高后下降的趋势,但在双价抗虫水稻中表现出随生育期逐渐下降的趋势。Cry1C蛋白在单、双价抗虫水稻的叶片组织中均表现出逐渐升高的趋势。与单价抗虫水稻相比,双价抗虫水稻中的Cry1Ab、Cry1C的蛋白表达水平并没有表现出显著的降低。因此,双价抗虫水稻不仅能高效防治害虫,还能延缓害虫抗性,在生产上表现出良好的应用前景。该研究结果可望为转Bt基因抗虫作物的环境安全评价提供科学数据和理论依据。     

Comparison of dose responses and resistance ratios in four populations of the rice stem borer, Chilo suppressalis (Lepidoptera: Pyralidae), to 20 insecticides

BACKGROUND: Chemical control is a major strategy for suppressing the rice stem borer, Chilo suppressalis (Walker). Owing to their high toxicity and increasing resistance development in the target insect, many insecticides will be phased out entirely in 2007 in China. Alternatives with relatively low toxicity are urgently needed to replace traditional chemicals for rice stem borer control. In this study, the authors examined four field populations of C. suppressalis for their toxicological responses to more than 20 insecticides, including a few low-toxicity organophosphates and many novel pesticides. Interpopulation resistance levels to 12 conventional insecticides were also compared. RESULTS: Based on LD(50) values, the rice stem borer was most sensitive to avermectins and fipronil (LD(50) < 1 ng larva(-1)). The stem borers exhibited the least sensitivity to endosulfan (LD(50) > 100 ng larva(-1)) and monosultap (LD(50) > 1000 ng larva(-1)). Insect growth regulators and chitin synthase inhibitors showed great efficacy against C. suppressalis, especially against populations that had developed resistance to conventional insecticides. Four field populations showed variable tolerance levels to many insecticides. LYG05 was the most susceptible population, only with a low level of resistance to monosultap (RR = 6.6). NC05 and GL05 populations exhibited intermediate tolerance levels with RR values up to 20.4 and 52.8 respectively. RA05 was the most resistant population to many insecticides, with resistance ratios up to 76.2. CONCLUSION: The results from this study provide valuable information for selection and adoption of new alternative insecticides and for resistance management of the rice stem borer.     

Biochemical evidence that an S431F mutation in acetylcholinesterase-1 of Aphis gossypii mediates resistance to pirimicarb and omethoate

Molecular changes in acetylcholinesterase (AChE) leading to target-site resistance to carbamate and organophosphate insecticides have recently been identified. Of particular interest is the S431F mutation in ace2 and its orthologue ace1 of Myzus persicae Sulzer and Aphis gossypii Glover, respectively. This mutation has been correlated with resistance to pirimicarb, but biochemical evidence has not yet been provided. Here, we describe for the first time that recombinantly expressed AChE1 from A gossypii carrying the S431F mutation is insensitive to pirimicarb and omethoate, but sensitive to demeton-S-methyl and hypersensitive to carbofuran. Furthermore, site-directed mutagenesis of the serine residue at position 431 in ace1 from a pirimicarb-susceptible clone of A gossypii conferred insensitivity to pirimicarb. We conclude that AChE1 of A gossypii is the target of toxicological relevance of carbamates and organophosphates.     

Cloning of the acetylcholinesterase 1 gene and identification of point mutations putatively associated with carbofuran resistance in Nilaparvata lugens

Molecular mechanisms of carbofuran resistance in the brown planthopper, Nilaparvata lugens Stål, were investigated. A carbofuran-resistant strain (CAS) showed approximately 45.5- and 15.1-fold resistance compared with a susceptible strain (SUS) and a non-selected field strain (FM), respectively. Activities of the esterase and mixed-function oxidase were approximately 2.8- and 1.6-fold higher, respectively, in the CAS strain than in the SUS strain, suggesting that these enzymes play a minor role in carbofuran resistance. Interestingly, the insensitivity of acetylcholinesterase (AChE) to carbofuran was approximately 5.5- and 3.7-fold higher in the CAS strain compared to the SUS and FM strains, respectively, indicating that AChE insensitivity is associated with carbofuran resistance. Western blot analysis identified two kinds of AChEs, of which the type-1 AChE (encoded from Nlace1, which is paralogous to the Drosophila AChE gene) was determined to be the major catalytic AChE in N. lugens. The open reading frame of Nlace1 is composed of 1989 bp (approximately 74 kD) and revealed 52.5% and 24.3% amino acid sequence identities to those of Nephotettix cincticeps and Drosophila melanogaster, respectively. Screening of point mutations identified four amino acid substitutions (G119A, F/Y330S, F331H and H332L) in the CAS strain that likely contribute to AChE insensitivity. The frequencies of these mutations were well correlated with resistance levels, confirming that they are associated with reduced sensitivity to carbofuran in N. lugens. These point mutations can be useful as genetic markers for monitoring resistance levels in field populations of N. lugens.     

Metabolism of imidacloprid and DDT by P450 CYP6G1 expressed in cell cultures of Nicotiana tabacum suggests detoxification of these insecticides in Cyp6g1-overexpressing strains of Drosophila melanogaster, leading to resistance

BACKGROUND: With the worldwide use of insecticides, an increasing number of pest insect species have evolved target-site or metabolism-based resistance towards some of these compounds. The resulting decreased efficacy of pesticides threatens human welfare by its impact on crop safety and further disease transmission. Environmental concentrations of some insecticides are so high that even natural populations of non-target, non-pest organisms such as the fruit fly Drosophila melanogaster Meig. have been selected for resistance. Cyp6g1-overexpressing strains of D. melanogaster are resistant to a wide range of chemically diverse insecticides, including DDT and imidacloprid. However, up to now there has been no evidence that the CYP6G1 enzyme metabolises any of these compounds. RESULTS: Here it is shown, by heterologous expression in cell suspension cultures of Nicotiana tabacum L. (tobacco), that CYP6G1 is capable of converting DDT (20 microg per cell culture assay) by dechlorination to DDD (18% of applied amount in 48 h), and imidacloprid (400 microg) mainly by hydroxylation to 4-hydroxyimidacloprid and 5-hydroxyimidacloprid (58 and 19% respectively in 48 h). CONCLUSION: Thus, the gap between the supposed resistance gene Cyp6g1 and the observed resistance phenomenon was closed by the evidence that CYP6G1 is capable of metabolising at least two insecticides.