二化螟抗药性监测及治理与高毒农药替代药剂筛选研究
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摘要
二化螟[Chilo suppressalis(Walker)]是危害我国水稻的主要害虫之一,近10年来,二化螟的发生为害迅速回升,严重阻碍了水稻的稳产和高产。目前化学防治仍是防治水稻二化螟的主要手段。原先在水稻上大量使用的5种有机磷类高毒农药如甲胺磷、对硫磷和甲基对硫磷等已于2007年1月1日起被全面禁止使用,且我国许多稻区二化螟已对常用治螟药剂杀虫单(双)和三唑磷等杀虫剂产生了高水平的抗性,并且近几年已监测到浙东南地区的田间二化螟种群对新型高效杀虫剂氟虫腈产生了中等水平抗性。因此,在防治二化螟方面,现在迫切需要寻找理想的药剂品种,来替代被禁用的高毒药剂以及替换或轮换二化螟已具有抗性的常用药剂。
本项研究旨在监测二化螟对常用药剂抗性的时空动态和研究田间二化螟种群抗药性进化机制,为二化螟抗性治理提供理论依据;测定田间二化螟对拟除虫菊酯类杀虫剂的敏感性和评估二化螟对菊酯类杀虫剂的抗性风险,为评估菊酯类杀虫剂是否可以在稻田中应用提供科学论据;针对目前高毒有机磷农药被禁用的政策,从现有杀虫剂中挑选出多种药剂,测定其对二化螟的室内毒力,以期选出理想的替代药剂作为推广部门进行大田药效示范试验的候选品种,再经天敌安全性及抗性风险评估,最终选出替代高毒农药的药剂品种。
为了监测我国田间二化螟对常用药剂的抗性时空动态,在2003～2006年期间采用点滴法测定了苏、浙、沪、赣和桂等五省(市、自治区)共9个二化螟田间种群对三唑磷、杀虫单、氟虫腈和阿维菌素等常用药剂的抗性水平。结果表明,所测种群对阿维菌素均处于敏感阶段(RR＜5)。除浙东南地区二化螟对氟虫腈具有中等水平抗性(2006年的RR为19.1～23.4倍)之外,其它种群对氟虫腈均敏感。不同地区二化螟对杀虫单和三唑磷产生了不同程度的抗性,浙东南地区种群对三唑磷和杀虫单分别产生了极高和高水平抗性;江苏高淳种群对三唑磷和杀虫单分别产生了高和极高水平抗性;江西南昌和广西桂林种群对三唑磷和杀虫单产生了中等至高水平抗性;上海青浦种群对三唑磷低抗和对杀虫单中抗;江苏常熟对三唑磷和杀虫单低抗;江苏连云港种群对杀虫单低抗和对三唑磷敏感。统计分析表明在2002～2006年的五年期间浙江瑞安种群对三唑磷和杀虫单抗性总体态势为上升趋势,对氟虫腈的抗性逐年上升。
为了评估菊酯类杀虫剂是否可以在稻田中应用,我们测定了田间二化螟种群对菊酯类杀虫剂的敏感性和进行了二化螟对菊酯类杀虫剂的抗性风险评估。在2004～2005年期间采用点滴法测定了江苏连云港和常熟、浙江瑞安及广西桂林四个二化螟田间种群对10种菊酯类杀虫剂的敏感性。结果表明,七种对鱼高毒菊酯类杀虫剂(高效氟氯氰菊酯、高效氯氟氰菊酯、高效氯氰菊酯、溴氰菊酯、S-氰戊菊酯、顺式氯氰菊酯及甲氰菊酯)＞三种对鱼低毒菊酯类杀虫剂(乙氰菊酯、醚菊酯和氟硅菊酯)＞甲胺磷和杀虫单。同时我们发现有些二化螟田间种群对某些菊酯类杀虫剂的敏感性正在明显降低,特别是瑞安种群二化螟对高效氟氯氰菊酯、溴氰菊酯的敏感性年度间变化最大(抗性倍数分别从2004年的16.2、10.6倍上升到2005年的131.4、60.4倍),这是国内外首次检测到二化螟对菊酯类杀虫剂产生抗性的报道(He et al.,2007)。另外发现瑞安种群对菊酯类杀虫剂的抗性与结构似乎有相关性,即对含α-氰基和乙烯基的环丙烷羧酸菊酯类杀虫剂产生高水平抗性,而对菊酸部分为异戊酸的氰戊菊酯很敏感,结合抗药性生化机制研究结果推测这种相关性可能与细胞色素P-450酶系的解毒代谢有关。
我们还进行了二化螟对乙氰菊酯和溴氰菊酯的抗性风险评估。乙氰菊酯筛选以2004年采自上海青浦地区二化螟(QP04)为起始种群,室内测定该种群对乙氰菊酯抗性水平很低(RR=5.6),筛选了8代后,抗性倍数上升到15.2倍,上升了1.72倍,估算所得的抗性现实遗传力h~2为0.1474,对其中的连续筛选4代(F_8～F_(11))进行估算所得h~2为0.2239。溴氰菊酯筛选以2005年采自江苏连云港地区二化螟(LYG05)为起始种群,室内测定该种群对溴氰菊酯很敏感(RR=2.2),筛选5代后,抗性倍数上升到5.8倍,上升了1.67倍,估算所得的抗性现实遗传力h~2为0.6618,对其中的连续筛选3代(F_0～F_2)进行估算所得h~2为1.4314。初步评估表明二化螟对乙氰菊酯的抗性风险明显低于对溴氰菊酯的抗性风险。
为了探究田间抗性二化螟对常用药剂抗性机制的生化特性,于2007年采用田间多抗性瑞安二化螟种群(RA07)为测试种群(对三唑磷、杀虫单、溴氰菊酯和氟虫腈的抗性水平分别为204.8、164.8、42.9和6.5倍),分别研究了脱叶磷DEF、顺丁烯二酸二乙酯DEM及增效醚PBO等三种增效剂对上述四种杀虫剂的增效作用。结果表明,PBO对溴氰菊酯有显著增效作用(增效系数为4.93倍),DEF和DEM对溴氰菊酯的毒力几乎无影响;DEF对三唑磷有弱增效作用(增效系数为1.38倍),DEM和PBO对三唑磷的毒力没影响;DEF、DEM和PBO对杀虫单和氟虫腈的毒力都几乎无影响。进一步对该种群二化螟的可能与抗性相关的酯酶和多功能氧化酶的离体活力进行测定,以室内敏感品系LYG05为参照品系发现,以α-醋酸萘酯为底物测定的RA07种群二化螟体内全酯酶的比活力是LYG05敏感品系二化螟的1.37倍,且差异显著。以对硝基苯醚为底物测定的RA07种群二化螟体内多功能氧化酶的比活力是LYG05敏感品系二化螟的1.74倍,且差异显著。研究结果表明多功能氧化酶活性增强是该种群对溴氰菊酯抗性的重要机理,酯酶可能是对三唑磷抗性的一个原因,但三种解毒代谢酶与对杀虫单和氟虫腈的抗性无关。
为了筛选出防治二化螟的高毒农药替代品种,我们于2005年采用点滴法测定了6类共22种供试药剂对江苏连云港、浙江瑞安、江西南昌和广西桂林四个具代表性的田间二化螟种群的毒力。发现不同类型的杀虫剂对四个种群的毒力次序大致类似,如阿维菌素类药剂和氟虫腈对二化螟的毒力最高,硫丹和杀虫单的触杀毒力最低,昆虫生长调节剂类杀虫剂(虫酰肼、呋喃虫酰肼、氟铃脲和氟啶脲)对二化螟具有较高的毒力,特别是对抗性二化螟仍表现出很高的杀虫活性。因此推荐氟虫腈、阿维菌素、甲氨基阿维菌素、虫酰肼、呋喃虫酰肼、氟铃脲、氟啶脲、哒嗪硫磷、喹硫磷、辛硫磷及毒死蜱等11种药剂作为候选品种供推广部门进行大田药效和示范试验的,结合天敌安全性及抗性风险评估,最终可选出替代5种高毒有机磷农药的药剂品种供推广应用。
The rice stem borer,Chilo suppressalis(Walker)(Lepidoptera:Pyralidae),is one of the economically important rice insects in China.In the last 10 years,population density and its damage intensity increased dramatically in China,which posed a serious threat to the trait of high and stable yield of the crop.Currently,control of C.suppressalis relies mainly on chemical insecticides.Five highly toxic organophosphates including methamidophos, parathion,methyl-parathion,monocrotophos,and phosphamidon will be banned in 2007 by the Ministry of Agriculture in China.In addition,populations of C.suppressalis in many rice regions of China have developed high levels of resistance to monosultap(bisultap) and triazophos,which are two additional conventional insecticides for the control of C. suppressalis.Resistance to a high efficacy novel insecticide fipronil has also been observed in some field populations in the last several years.Because of toxicity risk and increasing resistance development in C.suppressalis to conventional insecticides,it is urgent to screen available insecticides to find effective alternatives for replacing highly toxic OPs and other conventional insecticides.
The aims of present study include:to provide a foundation for resistance management of C.suppressalis,the dynamics of resistance of C.suppressalis to long-history-use insecticides such as monosultap and triazophos and the newly introduced insecticides fipronil and abamectin are monitored,and the possible underlying mechanisms of resistance to the conventional insecticides in field populations of C.suppressalis are studied; to provide scientific evidence for assess the feasibility of pyrethroids for rice insect control, the susceptibilities to pyrethroids in field populations of C.suppressalis are examined and the risk to develop resistance to pyrethroids in C.suppressalis is estimated;under the ban policy for the highly toxic Ops,a mass of compounds are chosen from insecticides in being, and are evaluated for their activities against C.suppressalis,in order to searching for candidate alternatives for field tests and for selecting additiveness and synergism of insecticide mixtures.
To monitor the dynamics of resistance of C.suppressalis,a total of eight populations were collected from 2003 to 2006 in five growing provinces in China and were used to examine their susceptibility levels to monosultap,triazophos,fipronil and abamectin using the topical application method.Results indicated that All 9 field populations collected in 2003～2006 were susceptible to abamectin(RR＜5).Most field populations were susceptible to fipronil(RR＜3),but the populations from Ruian and Cangnan,Zhejiang in 2006 showed moderate levels of resistance to fipronil(RR=19.1～23.4).Variable susceptibilities to triazophos and monosultap were detected in different populations.Those populations from the southeast Zhejiang province,had extremely high levels of resistance to triazophos and of high levels to monosultap;the population from Gaochun,Jiangsu had high levels of resistance to triazophos and of extremely high levels to monosultap;the two populations from Nanchang,Jiangxi and Guilin,Guangxi,evolved resistance of moderate or high levels to monosultap and triazophos;the population from Qingpu,Shanghai was low-level resistant to triazophos and moderate-level resistant to monosultap;the population from Changsu,Jiangsu had low levels of resistance to the two insecticides,and the population from Liangyungang,Jiangsu was low-level resistant to monosultap and susceptible to triazophos.The resistance levels in the RA population of rice stem borer exhibited an increasing trend(or with fluctuation) over a five-year period(2002～2006) for fipronil, monosultap and triazophos,and reached a maximal level in 2006 for all four insecticides.
To assess the feasibility of pyrethroids for rice insect control,we examined susceptibilities of four field populations(LYG,CS,RA and GL) of C.suppressalis to ten pyrethroids using the topical application method in laboratory in 2004 and 2005.Our results showed that the seven pyrethroids with high fish-toxicity(i.e.beta-cyfluthrin, lambda-eyhalothrin,beta-cypermethrin,deltamethrin,S-fenvalerate,alpha-cypermethrin, and fenpropathrin) were more effective against C.suppressalis than the three compounds with low fish-toxicity(i.e.cycloprothrin,etofenprox,and silafluofen).The results also showed that all ten of the pyrethroids were much more effective than methamidophos and monosultap for C.suppressalis control.In addition,we found that susceptibilities of some field populations of C.suppressalis to some high fish-toxicity pyrethroids were significantly reduced,and our results indicated that a Ruian(RA) field population showed a year-to-year variation in susceptibility to most tested pyrethroids between 2004 and 2005. Our data indicated that the tolerance levels increased dramatically in RA population, especially to beta-cyfluthrin and deltamethrin(with the resistance ratios from 16.2-and 10.6-fold,respectively,in 2004,to 131.4-and 60.4-fold in 2005,respectively).In addition, a close correlation between resistance ratios to the 10 compounds and differences of the structures of these compounds seems being established in the RA05 population,which was resistant to cyclopropane carboxylate pyrethroids with ethenyl and a-cyano while it was still very susceptible to the isovalerate pyrethroid fenvalerate with no cross resistance.This correlation is probably due to the detoxification mediated by the cytochrome P450-dependent monooxygenases(P450s),according to the study on biochemical resistance mechanism to insecticides in C.suppressalis described in Chapter 6.
Furthermore,the risk to develop resistance to pyrethroids in C.suppressalis is estimated.QP04 population was used to start cycloprithrin selection.It had resistance of a low level to triazophos before selection(RR=5.6).After 8 incontinuous selection cycles (generations),selected strain was 15.2-fold resistant to cycloprithrin,2.72 times higher than that pre-selection.Realized heritability(h~2)estimated from selection was 0.1474(0.2303 for F_8-F_(11),continuous selection).A similar deltamethrin selection experiment was initiated with LYG05 population,which was very susceptible to deltamethrin(RR=2.2).After 5 incontinuous selection cycles,deltamethrin resistance ratio of screened strain increased to 5.8-fold,2.67 times higher than that pre-selection,and the corresponding h~2 estimate was equal to 0.6618(1.4314 for F_0-F_2,continuous selection).It was preliminary indicated that this insect had a lower risk to evolve resistance to cycloprithrin than to develop resistance to deltamethrin.
To investigate the biochemical resistance mechanism to conventional insecticides in field population of C.suppressalis,synergism studies and detoxicating enzyme assays were conducted in 2007 on a field population of RA07,which was detected to be multi-resistant to triazophos(RR=204.8),monosultap(RR=164.8),pyrethroids(with 42.9-fold RR to deltamethrin),fipronil(RR=6.5) and etc.Synergism studies showed that piperonyl butoxide (PBO) had significant synergistic effect(4.93-fold) to deltamethrin,while S,S,S-tributyl phosphorotrithioate(DEF) and maleic acid diethyl ester(DEM) had no synergistic effect to deltamethrin;DEF had weak synergistic effect(1.38-fold) to triazophos,while DEM and PBO had no synergistic effect to triazophos;DEM,PBO and DEF had no synergistic effect to monosultap and fipronil.In vitro activity of esterase detected usingα-NA as a substrate and microsomal-O-demethylase using p-NA as a substrate showed significant differences between resistant(RA07) and susceptible(LYG05) strains with the increase ratios of 1.32-. and 1.74-fold,respectively.These results suggested that in this multi-resistance population, elevation in microsomal-O-demethylase metabolic activity was an importance mechanism of resistance to deltamethrin,esterase mediated detoxification might be a factor for resistance to triazophos,resistance to monosultap and fipronil had no close relations with three main metabolic enzymes(P450s,esterases and GSTs).
To provide alternative insecticides for the control of C.suppressalis,the topical application method was used to evaluate 22 insecticides which represented 6 different insecticide classes for their activities against four field populations of C.suppressalis with different resistance levels to conventional insecticides in 2005.Insecticides of different classes had the similar toxicity sequences in four populations.The rice stem borer was most sensitive to avermectins and fipronil,while C.suppressalis exhibited the least sensitivity to endosulfan and monosultap.Insect growth regulators(tebufenozide,JS118,hexaflumuron and chlorfluazuron) showed great efficacy against C.suppressalis,especially against the populations which have developed resistance to conventional insecticides.Efficacy data showed that seven novel insecticides(abamectin,emamectin bezoate,fipronil,tebufenozide, JS118,hexaflumuron and chlorfluazuron) and a few organophosphates(such as phoxim, chlorpyrifos,quinaphos and pyridaphenthion) were potential alternatives for replacing highly toxic pesticides for field trials.
本项研究旨在监测二化螟对常用药剂抗性的时空动态和研究田间二化螟种群抗药性进化机制,为二化螟抗性治理提供理论依据;测定田间二化螟对拟除虫菊酯类杀虫剂的敏感性和评估二化螟对菊酯类杀虫剂的抗性风险,为评估菊酯类杀虫剂是否可以在稻田中应用提供科学论据;针对目前高毒有机磷农药被禁用的政策,从现有杀虫剂中挑选出多种药剂,测定其对二化螟的室内毒力,以期选出理想的替代药剂作为推广部门进行大田药效示范试验的候选品种,再经天敌安全性及抗性风险评估,最终选出替代高毒农药的药剂品种。
为了监测我国田间二化螟对常用药剂的抗性时空动态,在2003～2006年期间采用点滴法测定了苏、浙、沪、赣和桂等五省(市、自治区)共9个二化螟田间种群对三唑磷、杀虫单、氟虫腈和阿维菌素等常用药剂的抗性水平。结果表明,所测种群对阿维菌素均处于敏感阶段(RR＜5)。除浙东南地区二化螟对氟虫腈具有中等水平抗性(2006年的RR为19.1～23.4倍)之外,其它种群对氟虫腈均敏感。不同地区二化螟对杀虫单和三唑磷产生了不同程度的抗性,浙东南地区种群对三唑磷和杀虫单分别产生了极高和高水平抗性;江苏高淳种群对三唑磷和杀虫单分别产生了高和极高水平抗性;江西南昌和广西桂林种群对三唑磷和杀虫单产生了中等至高水平抗性;上海青浦种群对三唑磷低抗和对杀虫单中抗;江苏常熟对三唑磷和杀虫单低抗;江苏连云港种群对杀虫单低抗和对三唑磷敏感。统计分析表明在2002～2006年的五年期间浙江瑞安种群对三唑磷和杀虫单抗性总体态势为上升趋势,对氟虫腈的抗性逐年上升。
为了评估菊酯类杀虫剂是否可以在稻田中应用,我们测定了田间二化螟种群对菊酯类杀虫剂的敏感性和进行了二化螟对菊酯类杀虫剂的抗性风险评估。在2004～2005年期间采用点滴法测定了江苏连云港和常熟、浙江瑞安及广西桂林四个二化螟田间种群对10种菊酯类杀虫剂的敏感性。结果表明,七种对鱼高毒菊酯类杀虫剂(高效氟氯氰菊酯、高效氯氟氰菊酯、高效氯氰菊酯、溴氰菊酯、S-氰戊菊酯、顺式氯氰菊酯及甲氰菊酯)＞三种对鱼低毒菊酯类杀虫剂(乙氰菊酯、醚菊酯和氟硅菊酯)＞甲胺磷和杀虫单。同时我们发现有些二化螟田间种群对某些菊酯类杀虫剂的敏感性正在明显降低,特别是瑞安种群二化螟对高效氟氯氰菊酯、溴氰菊酯的敏感性年度间变化最大(抗性倍数分别从2004年的16.2、10.6倍上升到2005年的131.4、60.4倍),这是国内外首次检测到二化螟对菊酯类杀虫剂产生抗性的报道(He et al.,2007)。另外发现瑞安种群对菊酯类杀虫剂的抗性与结构似乎有相关性,即对含α-氰基和乙烯基的环丙烷羧酸菊酯类杀虫剂产生高水平抗性,而对菊酸部分为异戊酸的氰戊菊酯很敏感,结合抗药性生化机制研究结果推测这种相关性可能与细胞色素P-450酶系的解毒代谢有关。
我们还进行了二化螟对乙氰菊酯和溴氰菊酯的抗性风险评估。乙氰菊酯筛选以2004年采自上海青浦地区二化螟(QP04)为起始种群,室内测定该种群对乙氰菊酯抗性水平很低(RR=5.6),筛选了8代后,抗性倍数上升到15.2倍,上升了1.72倍,估算所得的抗性现实遗传力h~2为0.1474,对其中的连续筛选4代(F_8～F_(11))进行估算所得h~2为0.2239。溴氰菊酯筛选以2005年采自江苏连云港地区二化螟(LYG05)为起始种群,室内测定该种群对溴氰菊酯很敏感(RR=2.2),筛选5代后,抗性倍数上升到5.8倍,上升了1.67倍,估算所得的抗性现实遗传力h~2为0.6618,对其中的连续筛选3代(F_0～F_2)进行估算所得h~2为1.4314。初步评估表明二化螟对乙氰菊酯的抗性风险明显低于对溴氰菊酯的抗性风险。
为了探究田间抗性二化螟对常用药剂抗性机制的生化特性,于2007年采用田间多抗性瑞安二化螟种群(RA07)为测试种群(对三唑磷、杀虫单、溴氰菊酯和氟虫腈的抗性水平分别为204.8、164.8、42.9和6.5倍),分别研究了脱叶磷DEF、顺丁烯二酸二乙酯DEM及增效醚PBO等三种增效剂对上述四种杀虫剂的增效作用。结果表明,PBO对溴氰菊酯有显著增效作用(增效系数为4.93倍),DEF和DEM对溴氰菊酯的毒力几乎无影响;DEF对三唑磷有弱增效作用(增效系数为1.38倍),DEM和PBO对三唑磷的毒力没影响;DEF、DEM和PBO对杀虫单和氟虫腈的毒力都几乎无影响。进一步对该种群二化螟的可能与抗性相关的酯酶和多功能氧化酶的离体活力进行测定,以室内敏感品系LYG05为参照品系发现,以α-醋酸萘酯为底物测定的RA07种群二化螟体内全酯酶的比活力是LYG05敏感品系二化螟的1.37倍,且差异显著。以对硝基苯醚为底物测定的RA07种群二化螟体内多功能氧化酶的比活力是LYG05敏感品系二化螟的1.74倍,且差异显著。研究结果表明多功能氧化酶活性增强是该种群对溴氰菊酯抗性的重要机理,酯酶可能是对三唑磷抗性的一个原因,但三种解毒代谢酶与对杀虫单和氟虫腈的抗性无关。
为了筛选出防治二化螟的高毒农药替代品种,我们于2005年采用点滴法测定了6类共22种供试药剂对江苏连云港、浙江瑞安、江西南昌和广西桂林四个具代表性的田间二化螟种群的毒力。发现不同类型的杀虫剂对四个种群的毒力次序大致类似,如阿维菌素类药剂和氟虫腈对二化螟的毒力最高,硫丹和杀虫单的触杀毒力最低,昆虫生长调节剂类杀虫剂(虫酰肼、呋喃虫酰肼、氟铃脲和氟啶脲)对二化螟具有较高的毒力,特别是对抗性二化螟仍表现出很高的杀虫活性。因此推荐氟虫腈、阿维菌素、甲氨基阿维菌素、虫酰肼、呋喃虫酰肼、氟铃脲、氟啶脲、哒嗪硫磷、喹硫磷、辛硫磷及毒死蜱等11种药剂作为候选品种供推广部门进行大田药效和示范试验的,结合天敌安全性及抗性风险评估,最终可选出替代5种高毒有机磷农药的药剂品种供推广应用。
The rice stem borer,Chilo suppressalis(Walker)(Lepidoptera:Pyralidae),is one of the economically important rice insects in China.In the last 10 years,population density and its damage intensity increased dramatically in China,which posed a serious threat to the trait of high and stable yield of the crop.Currently,control of C.suppressalis relies mainly on chemical insecticides.Five highly toxic organophosphates including methamidophos, parathion,methyl-parathion,monocrotophos,and phosphamidon will be banned in 2007 by the Ministry of Agriculture in China.In addition,populations of C.suppressalis in many rice regions of China have developed high levels of resistance to monosultap(bisultap) and triazophos,which are two additional conventional insecticides for the control of C. suppressalis.Resistance to a high efficacy novel insecticide fipronil has also been observed in some field populations in the last several years.Because of toxicity risk and increasing resistance development in C.suppressalis to conventional insecticides,it is urgent to screen available insecticides to find effective alternatives for replacing highly toxic OPs and other conventional insecticides.
The aims of present study include:to provide a foundation for resistance management of C.suppressalis,the dynamics of resistance of C.suppressalis to long-history-use insecticides such as monosultap and triazophos and the newly introduced insecticides fipronil and abamectin are monitored,and the possible underlying mechanisms of resistance to the conventional insecticides in field populations of C.suppressalis are studied; to provide scientific evidence for assess the feasibility of pyrethroids for rice insect control, the susceptibilities to pyrethroids in field populations of C.suppressalis are examined and the risk to develop resistance to pyrethroids in C.suppressalis is estimated;under the ban policy for the highly toxic Ops,a mass of compounds are chosen from insecticides in being, and are evaluated for their activities against C.suppressalis,in order to searching for candidate alternatives for field tests and for selecting additiveness and synergism of insecticide mixtures.
To monitor the dynamics of resistance of C.suppressalis,a total of eight populations were collected from 2003 to 2006 in five growing provinces in China and were used to examine their susceptibility levels to monosultap,triazophos,fipronil and abamectin using the topical application method.Results indicated that All 9 field populations collected in 2003～2006 were susceptible to abamectin(RR＜5).Most field populations were susceptible to fipronil(RR＜3),but the populations from Ruian and Cangnan,Zhejiang in 2006 showed moderate levels of resistance to fipronil(RR=19.1～23.4).Variable susceptibilities to triazophos and monosultap were detected in different populations.Those populations from the southeast Zhejiang province,had extremely high levels of resistance to triazophos and of high levels to monosultap;the population from Gaochun,Jiangsu had high levels of resistance to triazophos and of extremely high levels to monosultap;the two populations from Nanchang,Jiangxi and Guilin,Guangxi,evolved resistance of moderate or high levels to monosultap and triazophos;the population from Qingpu,Shanghai was low-level resistant to triazophos and moderate-level resistant to monosultap;the population from Changsu,Jiangsu had low levels of resistance to the two insecticides,and the population from Liangyungang,Jiangsu was low-level resistant to monosultap and susceptible to triazophos.The resistance levels in the RA population of rice stem borer exhibited an increasing trend(or with fluctuation) over a five-year period(2002～2006) for fipronil, monosultap and triazophos,and reached a maximal level in 2006 for all four insecticides.
To assess the feasibility of pyrethroids for rice insect control,we examined susceptibilities of four field populations(LYG,CS,RA and GL) of C.suppressalis to ten pyrethroids using the topical application method in laboratory in 2004 and 2005.Our results showed that the seven pyrethroids with high fish-toxicity(i.e.beta-cyfluthrin, lambda-eyhalothrin,beta-cypermethrin,deltamethrin,S-fenvalerate,alpha-cypermethrin, and fenpropathrin) were more effective against C.suppressalis than the three compounds with low fish-toxicity(i.e.cycloprothrin,etofenprox,and silafluofen).The results also showed that all ten of the pyrethroids were much more effective than methamidophos and monosultap for C.suppressalis control.In addition,we found that susceptibilities of some field populations of C.suppressalis to some high fish-toxicity pyrethroids were significantly reduced,and our results indicated that a Ruian(RA) field population showed a year-to-year variation in susceptibility to most tested pyrethroids between 2004 and 2005. Our data indicated that the tolerance levels increased dramatically in RA population, especially to beta-cyfluthrin and deltamethrin(with the resistance ratios from 16.2-and 10.6-fold,respectively,in 2004,to 131.4-and 60.4-fold in 2005,respectively).In addition, a close correlation between resistance ratios to the 10 compounds and differences of the structures of these compounds seems being established in the RA05 population,which was resistant to cyclopropane carboxylate pyrethroids with ethenyl and a-cyano while it was still very susceptible to the isovalerate pyrethroid fenvalerate with no cross resistance.This correlation is probably due to the detoxification mediated by the cytochrome P450-dependent monooxygenases(P450s),according to the study on biochemical resistance mechanism to insecticides in C.suppressalis described in Chapter 6.
Furthermore,the risk to develop resistance to pyrethroids in C.suppressalis is estimated.QP04 population was used to start cycloprithrin selection.It had resistance of a low level to triazophos before selection(RR=5.6).After 8 incontinuous selection cycles (generations),selected strain was 15.2-fold resistant to cycloprithrin,2.72 times higher than that pre-selection.Realized heritability(h~2)estimated from selection was 0.1474(0.2303 for F_8-F_(11),continuous selection).A similar deltamethrin selection experiment was initiated with LYG05 population,which was very susceptible to deltamethrin(RR=2.2).After 5 incontinuous selection cycles,deltamethrin resistance ratio of screened strain increased to 5.8-fold,2.67 times higher than that pre-selection,and the corresponding h~2 estimate was equal to 0.6618(1.4314 for F_0-F_2,continuous selection).It was preliminary indicated that this insect had a lower risk to evolve resistance to cycloprithrin than to develop resistance to deltamethrin.
To investigate the biochemical resistance mechanism to conventional insecticides in field population of C.suppressalis,synergism studies and detoxicating enzyme assays were conducted in 2007 on a field population of RA07,which was detected to be multi-resistant to triazophos(RR=204.8),monosultap(RR=164.8),pyrethroids(with 42.9-fold RR to deltamethrin),fipronil(RR=6.5) and etc.Synergism studies showed that piperonyl butoxide (PBO) had significant synergistic effect(4.93-fold) to deltamethrin,while S,S,S-tributyl phosphorotrithioate(DEF) and maleic acid diethyl ester(DEM) had no synergistic effect to deltamethrin;DEF had weak synergistic effect(1.38-fold) to triazophos,while DEM and PBO had no synergistic effect to triazophos;DEM,PBO and DEF had no synergistic effect to monosultap and fipronil.In vitro activity of esterase detected usingα-NA as a substrate and microsomal-O-demethylase using p-NA as a substrate showed significant differences between resistant(RA07) and susceptible(LYG05) strains with the increase ratios of 1.32-. and 1.74-fold,respectively.These results suggested that in this multi-resistance population, elevation in microsomal-O-demethylase metabolic activity was an importance mechanism of resistance to deltamethrin,esterase mediated detoxification might be a factor for resistance to triazophos,resistance to monosultap and fipronil had no close relations with three main metabolic enzymes(P450s,esterases and GSTs).
To provide alternative insecticides for the control of C.suppressalis,the topical application method was used to evaluate 22 insecticides which represented 6 different insecticide classes for their activities against four field populations of C.suppressalis with different resistance levels to conventional insecticides in 2005.Insecticides of different classes had the similar toxicity sequences in four populations.The rice stem borer was most sensitive to avermectins and fipronil,while C.suppressalis exhibited the least sensitivity to endosulfan and monosultap.Insect growth regulators(tebufenozide,JS118,hexaflumuron and chlorfluazuron) showed great efficacy against C.suppressalis,especially against the populations which have developed resistance to conventional insecticides.Efficacy data showed that seven novel insecticides(abamectin,emamectin bezoate,fipronil,tebufenozide, JS118,hexaflumuron and chlorfluazuron) and a few organophosphates(such as phoxim, chlorpyrifos,quinaphos and pyridaphenthion) were potential alternatives for replacing highly toxic pesticides for field trials.
引文
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