邯郸地区棉铃虫抗药性的一些生物化学特性(英文)

对河北邯郸地区的抗性棉铃虫(Helicovrpa armigera H(?)bner)种群不同发育阶段乙酰胆碱酯酶(AChE)对灭多威和对氧磷的敏感磷的敏感度结果表明:4-5龄幼虫的AChE在棉铃虫整个发育期中敏感度是最低的,成虫期的AChE最敏感,蛹期AChE平均敏感度低于幼虫期。用甲基对硫磷和灭多威以LD_5的诱导剂量和LD_(50)的选择剂量处理棉铃虫幼虫后,AChE对底物的亲和力和对抑制剂的敏感度发生了明显的变化,水解β-乙酸萘酯的羧酸酯酶对底物的亲和力明显提高,而水解α-乙酸萘酯的羧酸酯酶对底物的亲和力在处理后24小时无明显改变。谷胱甘肽-S-转移酶活性,低剂量诱导组最高降低至78％和43％,而高剂量选择组提高15％,诱导组和选择组对底物的亲和力均明显降低,处理的剂量和处理后时间明显地影响棉铃虫磷酸酯酶的活性及其对底物的亲和力。     

磷酸酯酶与乙酰胆碱酯酶在棉铃虫对辛硫磷抗药性中的作用(英文)

利用人工选育的方法 ,获得了棉铃虫对辛硫磷的抗药性品系和相对敏感品系 ,其对辛硫磷的LD50分别为 1 1 .0 574μg/虫和 0 .9790 μg/虫。以生物测定与生化测定的方法 ,比较了两品系的差异。增效剂活体试验测定结果表明 ,磷酸三苯酯、增效磷对棉铃虫三龄幼虫均表现出一定的增效作用 ,但在两品系间的增效作用存在差异 ,对相对敏感品系的增效倍数分别为 2 .2 7倍、2 .1 3倍 ,对抗性品系的增效倍数分别为 6.93倍和 6.43倍。磷酸酯酶活性测定结果表明 ,抗性品系的酶活力分别是相对敏感品系的1 6.67倍 (碱性 )和 1 .89倍 (酸性 ) ,Km 和Vmax则分别为 0 .76倍、2 .64倍 (碱性 )和 1 .38倍、1 .78倍 (酸性 ) ;而羧酸酯酶的活性则差异不大。对靶标酶乙酰胆碱酯酶的活性测定表明 ,与相对敏感品系相比 ,抗性品系酶活性显著增强 ,并且对杀虫剂的亲和力降低 ,说明乙酰胆碱酯酶在棉铃虫对辛硫磷的抗药性中也有着重要的作用     

棉铃虫AChE不同分子型对抑制剂的敏感度及其与抗药性的关系(英文)

在分离到的棉铃虫AChE五种不同的分子型中 ,2 .1s和 8.7sAChE抗性品系对毒扁豆碱的敏感度明显低于敏感品系 ,成虫头部I50 值分别相差 1 86.3和 84.8倍 ,幼虫I50 值分别相差 1 0 1 0 倍和 1 0 5 倍。幼虫 5.3sAChE对毒扁豆碱的敏感度抗性品系和敏感品系相差达 1 2 3倍 ,而成虫则没有差异。研究结果表明 2 .1s、5.3s和 8.7sAChE敏感度降低可能是造成棉铃虫对有机磷和氨基甲酸酯类杀虫药剂产生抗性的主要原因     

棉铃虫的细胞色素P-450单加氧酶系与菊酯类杀虫剂抗药性的关系(英文)

细胞色素Ｐ－４５０单加氧酶系在昆虫抗药性中起重要作用。本文研究了棉铃虫抗性品系与敏感品系间细胞色素Ｐ４５０含量、对－硝基茴香醚Ｏ－脱甲基酶和艾氏剂环氧化酶的差异。结果表明：抗性品系中细胞色素Ｐ－４５０含量、对－硝基茴香醚Ｏ－脱甲基酶活性分别是敏感品系的１．７１倍和２．２１倍,而艾氏剂环氧化酶活性仅为１．３５倍。因此细胞色素Ｐ－４５０含量和对－硝基茴香醚Ｏ－脱甲基酶在棉铃虫抗菊酯品系中起重要作用。进一步讨论了不同抗性品系间抗性机制不同的原因。     

棉铃虫种群遗传变化的研究(英文)

利用重复序列引物扩增DNA多态性方法研究了几个产棉区的棉铃虫的种群遗传结构变化。结果表明 :实验室种群比自然种群的遗传变化低 ,自然种群间的遗传差异不显著 ,同一地点不同年间种群间存在较大遗传差异 ,表明棉铃虫的迁飞扩散现象比较明显 ;聚类分析结果表明 ,来自辽宁的朝阳种群 (CY)与来自山东高唐 (GT)、江苏大丰 (DF)的种群间的有较强的基因流 ,为我国东北地区棉铃虫主要来自河北、山东等地的假说提供了一定的理论依据。本研究所选用的遗传标记能灵敏地检测种群的遗传结构变化     

彭泽县棉铃虫抗药性监测

棉铃虫Helicoverpaarmigera(Hubner）是棉花生长中、后期的一种主要害虫,其抗药性问题已成为棉花生产主要障碍之一[1]。为了解我地棉铃虫的抗药性情况,我们自1996年开始对其进行监测。现将主要结果整理如下。1材料与方法1．1测定药剂10％氯氰菊酯乳油、2．5％敌杀死乳油、90％万灵可溶性粉剂、35％硫丹乳油、2．5％功夫乳油。药剂由全国农技推广中心提供。氯氰菊酯、万灵测定其抗性倍数,硫丹、功夫和敌杀死测定一个浓度（硫丹稀释800倍,功夫和敌杀死各稀释1500倍）的毒杀效果,重复3次取平均值。回．2测试虫源在全县不同棉区采集第三代…     

昆虫抗药性和昆虫毒理动力学(英文)

不断地使用一种杀虫药剂防治昆虫,会导致昆虫产生抗药性。对昆虫抗药性资料进行广泛综述时,发现了仅单独的解毒作用不能被解释为家蝇对有机氯杀虫药剂产生高抗性原因。作为一个基因。家蝇可以对有机氯产生比对有机磷杀虫剂更高的抗药性,尽管有机磷杀虫剂一般在虫体内是不太稳定的。考虑到昆虫毒理的动力学,杀虫药剂的穿透作用更显示出其实际的重要性。根据穿透和解毒的速率,慢的穿透作用是解毒作用的一个限制因子。防治敏感和抗性昆虫的观察结果,可以划出物理和生物因子之间关系的几种相关曲线图解。这些相关性不仅能说明家蝇对有机磷和有机氯杀虫剂的抗性程度,而且也助于选择出新的杀虫毒剂。     

中国北方棉蚜对杀虫剂的抗药性(英文)

棉蚜(Aphis gossypu Glover)是我国北方棉区的主要害虫。自1953年开始使用有机磷杀虫剂对硫磷和1059等防治棉蚜以来,田间喷雾浓度和使用次数逐年增大,棉蚜产生的抗性。80年代使用高效拟除虫菊酯类杀虫剂溴氰菊酯和氰戊菊酯等,三年后棉蚜抗性增长了171倍,有些地区达3230倍。此文报道棉蚜的抗性机制的研究结果,证实了棉蚜抗性与杀虫剂穿透能力降低、多功能氧化酶、α-乙酸萘酯酶和α-乙酸萘酯羧酸酶的活力增加、乙酰胆碱酯酶对杀虫剂敏感性的降低有关。在这基础上进行增效剂SV_1对多种有机磷和拟除虫菊酯杀虫剂的增效试验,得出SV_1对防治抗性棉蚜有明显的增效作用,能延缓抗性的发展。并研究了杀虫剂抗性的防治策略及综合治理措施,为防治棉蚜抗性和延缓抗性发展取得了较好的效果。     

棉铃虫前胸腺的神经解剖研究(英文)

用光学显微镜和电子显微镜对棉铃虫(Helicoverpa armigera)前胸腺的形态、超微结构、神经分布及末龄幼虫和早期蛹前胸腺的超微结构变化作了观察和研究。前胸腺呈T形,每对由76—116个大小不同的腺细胞组成;在末龄腺体中发现一种直径约6μ的微腺细胞;在28C,光周期L:D=15:9条件下,前胸腺在化蛹后第三天完全消失,前胸腺布满大量的气管或微气管,并有三根神经分布。腺细胞质膜内陷形成细胞间隙系统(ICS):ICS的宽度和深度在末龄幼虫蜕皮甾类分泌达到峰值时增至最大。在第4天末龄幼虫的前胸腺中观察到大量的多泡囊体(MVS)及其残片、粗面内质网、线粒体基质的电子透性增至最大。最后,对第十天末龄幼虫前胸腺细胞间边的冷冻复型膜进行了观察。前胸腺结构的变化与其分泌状态密切相关。     

棉铃虫一种新的实用人工饲料(英文)

本文报道饲养棉铃虫的一种新的含有番茄酱的实用人工饲料。取食这种饲料时,绝大部分幼虫只有5个龄期。因此,幼虫历期大为缩短。在该饲料中加入适量绿豆粉取代麦胚能在一定程度上提高成虫的繁殖力。在连续12代饲养期间,幼虫期存活率在87．5％－98．7％之间,蛹期死亡率不到5％;第11代成虫平均产卵千粒左右,孵化率达86．5％。这些结果表明,经过2年连续饲养后,棉铃虫的活力并没有明显的降低。     

SINGLE-HELICOVERPA ARMIGERA TEST TO DETERMINE INSECTICIDE RESISTANCE GENOTYPE FREQUENCY*

Abstract A sensitive technique for identifying insecticide resistance genotype frequency of the Ace gene existing in natural populations of Helicoverpa armigera in China is described. The technique is based on the comparison of acetylcholinesterase (AChE) activity in 3 equal aliquots, which are taken from the homogenate of a single H. urnzigera: 1) in absence of inhibitor (well No. 1); 2) in absence of enzyme precursor (well No. 2); and 3) in presence of a concentration of propoxur inhibiting the AChE coded by the Ace^Sallele but not by the Ace^Rallele (well No. 3). An intensity of same strength in wells No. 1 and No. 3 indicates that propoxur has no effect on AChE activity, and the genotype should be Ace^RR(when well No. 3 = well No. 1). A same weak intensity in well No. 2 and No. 3 (and a strong intensity in well No. l), indicates that propoxur completely inhibited AChE activity; and the genotype should be Ace^Ss(when well No. 3 = well No. 2). Well No. 3 displaying an intermediate intensity between well No. 1 (strong intensity) and well No. 2 (weak intensity) indicates that AChE activity was partially inhibited by the propoxur concentration and genotype should be Ace^Rs(when well No. 2<well No. 3<well No. 1). The genotype frequency was determined in 1995 and 1996. the Ace^SSis 76. 1%, 70. 6% for low resistance populations respectively whereas the Ace^RRis 68. 3%, 65. 3% for high resistance population respectively.     

RESISTANCE ALLEVIATION IN THE LARVAL COTTON BOLLWORM TO FENVALERATE AFTER PRE-TREATMENT WITH BACILLUS THURINGIENSIS

Abstract  Resistance alleviation and mechanism in the 2nd instar larvae of one susceptible and two resistant strains of the cotton bollworm, Helicoverpa armigera to fenvalerate were studied at 24 h after the larvae were treated with Bacillus thuringiensis Berliner ( B.t. ) preparation. The results showed that responses of larvae with B. t pretreatment to fenvalerate were much more sensitive than those without B. t pretreatment. Compared with the susceptible strain (YZ-S), net ratios between LD₅₀s with and without B. t pretreatment in the two resistant strains increased approximate 62. 6% and 80. 9%, respectively) inhibition of specific inhibitors to esterases displayed that efficacy of malaoxon and paraoxon methyl to acetyl-cholinesterase increased from 66.1% to 99. 8% in vitro and in vivo ; dynamic factors of acetylcholinesterase were significantly changed, in which K _m and V _max values decreased from 45. 58% to 68. 62% and K _i values of malaoxon and paraoxon methyl to acethylcholinesterase increased approximate 60%. It suggested that the sensitivity of the 2nd instar larvae of resistant H. armigera to fenvalerate might increase after B. t pretreatment for 24 h, and change of acetylcholinesterase was an important factor during resistance alleviation by B. t pretreatment. The research showed that it is practicable to bring the coordinated use of B. t and chemical insecticides in IPM system.     

CYTOCHROME P450 MONOOXYGENASES ASSOCIATED WITH PYRETHROID RESISTANCE IN HELICOVERPA ARMIGERA (HÜBNER)

Abstract  The relationship between the cytochrome P-450 (cytP⁴⁵⁰ monooxygenase and insecticide resistance mechanisms in Helicoverpa armigera was studied. The level of cytP450 and activity of p -ni-troanisole ( p -NA) O-demethylase in resistant strain were 1. 71 times and 2. 21 times respectively higher as compared with those of the susceptible strain. Aldrin epoxidase activity showed 1. 35 fold strain difference. Therefore aldrin epoxidase might not be important for H. armigera resistance mechanism, the level of cytP450 and activity of p -NA O-demethylase might play a vital role in H. armigera resistance mechanisms. The reason of different resistance mechanisms among different H. armigera resistant strains is discussed.     

苏云金杆菌预处理棉铃虫二龄幼虫对氰戊菊酯抗性缓解及机理研究(英文)

本文研究了Ｂ．ｔ预处理棉铃虫二龄幼虫对氰戊菊酯抗性缓解及其机理。结果表明,Ｂ．ｔ预处理后的幼虫对氰戊菊酯的敏感性提高。与敏感性种群相比较,在Ｂ．ｔ预处理和未处理两个抗性种群的幼虫对氰戊菊酯的ＬＤ_５０之间纯比值分别大约增加了６２．６％和８０．９％。特异性酶抑制剂对乙酰胆碱酯酶的抑制作用显示,在活体内,马拉氧磷和甲基对氧磷对乙酰胆碱酯酶抑制效率提高了６６．１％和９９．８％;在活体外抑制效率（Ｉ_５０） ６０％和６７．５％及５５．６５％和８６．４％。同时,乙酰胆碱酯酶动力学各因子变化显著。在Ｂ．ｔ预处理的抗性棉铃虫幼虫中,Ｋｍ值从４５．５８％减少到６８．６２％;Ｋｉ值分别增加了２１．２％和４０．９％及３８．２％和６１．５％。这些结果证实Ｂ．ｔ预处理２４ｈ抗性棉铃虫二龄幼虫对氰戊菊酯的敏感性提高,乙酸胆碱酯酶的变化是重要原因之一。这一研究为在ＩＰＭ系统中将微生物杀虫剂和化学杀虫剂协调使用缓解棉铃虫对拟除虫菊酯类杀虫剂的抗性提供理论基础。     

BIOCHEMICAL ASPECTS OF INSECTICIDE RESISTANCE IN COTTON BOLLWORM FROM HANDAN OF HEBEI PROVINCE*

Abstract The cotton bollworms, Helicoverpa armigera Hübner, collected from Handan of Hebei Province, have evolved high resistance to pyrethroid, organophosphate and carbamate insecticides, The sensitivity of acetylcholinesterase (AChEs) to paraoxon and methomyl varied with the development stages of the cotton bollworm. After the treatments with LD₅ and LD₅₀ of parathion and methomyl to the cotton bollworms, the affinity of AChE to acetylthiocholine (ATCh) and acetyl-β-bmethyl-thio choline (MeTCh) increased significantly except the treatment of parathion using LD₅₀ dosage while the sensitivity of AChEs to paraoxon significantly decreased. The sensitivity of AChEs to methomyl strongly increased in the treatment of parathion using LD₅₀ dosages while strongly decreased in other treatments. The affinity of carboxylesterase to β-naphthyl acetate (β-NA) was higher in groups of treatment with insecticides than in group of control. The glutathione-s-transferase (GST) activity significantly decreased in the induced groups using LD₅ dosages, while increased in the selection groups using LD₅₀ dosages. The effects of parathion and methomyl on the phosphatases of cotton bollworm were related to the dosages of application and the time after treatment and the effect on the alkaline phosphatase was stronger than on acid phosphatase.     

不同抗性水平棉铃虫地理种群的基因交流对抗性演化的影响

采自我国长江流域棉区的棉铃虫Helicoverpa armigera(Hubner)四川简阳种群、湖北武穴种群、湖南衡阳种群、江苏南京种群和浙江杭州种群对辛硫磷、三氟氯氰菊酯和硫丹的抗性水平分别为8.05～38.80、2.86～290.00和5.87～19.83倍。抗性较高的简阳种群和较低的杭州种群正、反交F₁代对上述3种农药的抗性为13.16和16.69倍、10.43和19.00倍及12.42和10.21倍,抗性遗传的显性系数为-0.2220、-0.2147和0.0700,表明对辛硫磷、三氟氯氰菊酯抗性呈不完全隐性方式遗传,而硫丹抗性则为半隐性遗传。P1自交F2代对辛硫磷、三氟氯氰菊酯和硫丹的抗性水平分别为17.66、7.57和9.03倍,F₁代对杭州种群回交F₂代的抗性倍数分别为8.79、3.Oo和6.61。其结果,高抗区和低抗区棉铃虫种群的基因流动将导致对三种农药抗性水平明显下降。     

MOLECULAR MECHANISMS OF TARGET RESISTANCE IN THE ORGANOPHOSPHATE- AND PYRETHROID-RE-SISTANT HELICOVERPA ARMIGERA (HUBNER)

Abstract  The molecular mechanisms of target resistance, i. e. acetyicholinesterase (AChE) and sodium channel insensitivity, in the organophosphate(0P)- and pyrethroid(Py)-resistant (R) Helicoverpa armigera were investigated. The activity and V_max of AChE from R strain were 1.09– and 1.23-fold of the susceptible(S) strain respectively, but the K_M value of AChE in R strain was only 0.67-fold of S stain. The K_i values of AChE from the R strain were 0.44 for DDVP and 0.50 for malathion respectively, as compared with those of the S strain. These data showed that the AChE from R strain might be qualitatively altered. The knockdown resistance ( kdr ) of the resistant H. armigera was also identified by PCR technique. The fragments of IIS6, linker of 11–111 and a part of IIS5-IIs6 in the sodium channel were cloned and sequenced, and then compared to amino acid sequences from the R and S strains. and other insect species. There was no difference to be found at amino acid level in the above fragments cloned. It was suggested that the Rdr was not involved in the resistance of R strain.     

SUBSTRATE SPECIFICITY AND DEVELOPMENTAL CHANGES OF ACETYLCHOLINESTERASE IN COTTON BOLLWORM*

Abstract The substrate specificity and developmental changes of acetylcholinesterase (AChE) of cotton bollworm, Helicoverpa armigera Hübner, were investigated during 1991 to 1994. The insects were collected from Handan suburbs of Hebei Province and Guan County of Shandong Province. The results show that the specific activity and Michaelis constants (k_m) of AChE toward acetylthiocholine (ATCH) and acetyl-β-methyl-thiocholine (MeTCh) regularly varied with the developmental stage of cotton bollworm. The two peaks of the specific activity were observed respectively in the third instar and sixth instar of larvae. The specific activity of AChE in pupae was the lowest and that in heads of four-days moth was the highest in various developmental stages of cotton bollworms. The tendency of K_m and maximum velocity (V_max) was identical with the change of specific activity in the AChEs of cotton bollworm. The activation energy (Ea) of AChE toward MeTCh in pupae and adults was 3. 9–4. 3 times as much as that of, larvae in cotton bollworms from Handan of Hebei Province. It suggests that the spending energies of AChE for hydrolysing substrate are different in larva, pupa and adult. The optimum conditions of AChE reaction with ATCh in larvae were 50–100 mg of tissue weights for the amount of enzyme, 10–20 minutes for the reaction time, 35°C for the reaction temperature and 8. 0 for the reaction pH value.     

棉铃虫乙酰胆碱酯酶(AChE)的体躯分布及部分纯化

该文研究了棉铃虫Helicoverpa armigera乙酰胆碱酯酶(AChE)的体躯分布及部分纯化技术。在幼虫和成虫体内均以头部AChE活性最高;AChE在成虫和幼虫头、胸、腹部的活性分布分别为78.2%、3.9%、17.9%和90.3%、6.5%和3.2%。通过用Sepharose 4B和Sephadex G-200纯化后,AChE的最高纯化倍数为3.66和17.74倍。样品和洗脱液中含有Triton X-100时,纯化倍数明显高于无Triton X-100的处理组。