Changes in susceptibility to conventional insecticides of a Cry1Ac-selected population of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

The changes in the susceptibility of the cotton bollworm, Helicoverpa armigera (Hübner) to three insecticides (lambda-cyhalothrin, phoxim and endosulfan) commonly used for control of this pest in China were monitored by bioassays at various generations. The insects were originally collected from Bt cotton fields and selected with Cry1Ac over 44 generations. In comparison with a susceptible strain, the larval resistance of the Bt-selected populations to Cry1Ac toxin increased 106-fold. Simultaneously, the resistance levels to lambda-cyhalothrin, phoxim and endosulfan declined dramatically. The results indicated no positive cross-resistance between Cry1Ac toxin and the insecticides. Evidence of the lack of cross-resistance to three commonly used synthetic insecticides in our laboratory-derived Cry1Ac-resistant population may suggest that growers can confidently use these insecticides if and when resistance to Cry1Ac cotton does occur.     

转Bt基因棉对甜菜夜蛾的影响

随着转基因棉在全球的广泛种植,棉铃虫(Helicoverpa armigera)等靶标害虫得到了有效控制,但是甜菜夜蛾的为害呈现日益加重的趋势,并从次要害虫上升为主要害虫。本文论述了Bt毒蛋白对甜菜夜蛾生长发育、取食行为、体内酶活、药剂敏感性等的影响以及甜菜夜蛾对Bt毒蛋白产生抗性的机制等,并对今后转基因棉田甜菜夜蛾的防治提出了建议。     

华北和新疆棉铃虫对三类农药抗性发展趋势分析

1996－1999年监测了河北冀州,河南西华,山东高密和新疆库尔勒棉铃虫对氯氟氰菊酯,久效磷和灭多威的抗性动态变化,结果表明,新疆棉铃虫抗性水平变化很少,尚处于敏感状态或低抗水平,而华北棉铃虫已处于中到高抗水平,不同年份比较,对氯氟氰菊酯,冀州棉铃虫抗性呈下降趋势,而西华和高密棉铃虫在1996－1998年呈上升趋势,但自1998年起华北地区大量种植Bt棉,使得1999年棉铃虫抗性有所下降,对久效磷和灭多威,华北各地棉铃虫的抗性在1996－1999年均呈上升趋势。棉铃虫抗性个体频率变化趋势与上述结果基本一致。     

棉铃虫抗药性监测及其化学防治

1997～1998年,用浸叶接虫法测定了山西省南部棉区永济县棉铃虫对常用5种农药的抗药性,同时进行了多种农药防治棉铃虫田间药效试验研究。结果表明,永济县棉铃虫对氯氰菊酯已产生了高等水平的抗药性,对久效磷产生了中等水平的抗药性,但对灭多威仍未产生明显的抗药性。水胺硫磷、甲基对硫磷、氯氟氰菊酯、氯氰菊酯、丙·辛乳油和甲对·氯乳油等农药防治永济县二代棉铃虫效果较好     

转Bt基因棉种植前后3个棉铃虫种群对4种杀虫剂的抗性变化

分别于1990年、1992年、2001年和2005年从山东泰安、山东夏津和湖北天门采集3个棉铃虫H elicoverpa arm igera种群,采用点滴法测定了4种杀虫剂对其的毒力,以监测各种群的抗药性变化趋势。结果表明:19901992年3个种群棉铃虫对氰戊菊酯的抗性增长迅速(13.256.1倍);19922001年抗性增长速度有所减缓(3.111.0倍);20012005年抗性水平降低。3个种群棉铃虫对灭多威和辛硫磷的抗性变化趋势与氰戊菊酯的基本相似,但抗性水平较低;对硫丹则均一直处于敏感水平。引起抗药性变化的原因主要与各类药剂的使用频率和转B t基因抗虫棉的大面积推广有关。     

棉铃虫Bt毒素受体蛋白—氨肽酶N与抗性的关系

随着我国转基因棉花种植面积的日益增加,主要目标害虫—棉铃虫的抗性问题越来越被大家所关注。氨肽酶N(aminopeptidase N,APN)是一类昆虫中肠内Bt毒素的受体蛋白,其结构、结合位点的改变或基因突变可能是昆虫对Bt毒素产生抗性的重要原因。本文通过分析Bt毒素的作用方式,从生化、生理、分子生物学等方面探讨了棉铃虫Bt毒素受体蛋白APN与抗性的关系。     

棉铃虫围食膜中Bt抗性相关蛋白的分离与鉴定

棉铃虫是世界性的重要农业害虫。围食膜作为昆虫抵御病原微生物入侵及有害物质的第一道天然保护性屏障,其上可能存在与Bt抗性相关的受体蛋白。本研究以Bt Cry1Ac抗性和敏感品系的棉铃虫围食膜为对象,采用NuPAGE电泳技术、配体杂交、质谱鉴定和生物信息学分析等技术,测定了围食膜蛋白含量,鉴定了蛋白质的组成及与Bt Cry1Ac毒素的结合能力。结果表明敏感品系围食膜中蛋白含量为22.19%,抗性品系围食膜中蛋白含量为26.99%。抗、感品系棉铃虫围食膜上存在与Cry1Ac毒素结合的6个差异蛋白,推测其中棉铃虫羧酸酯酶蛋白和血影蛋白是2个有意义的抗性相关蛋白,2个新蛋白可能参与Bt抗性。研究证明棉铃虫围食膜上存在Bt结合蛋白且与抗性相关,为进一步明确Bt抗性机制、制定合理的Bt抗性治理策略提供了理论依据。     

Long-term selection for resistance to transgenic cotton expressing Bacillus thuringiensis toxin in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

Selection experiments for resistance to transgenic Bt cotton expressing Cry1Ac toxin in Helicoverpa armigera (Hübner) were conducted using a leaf-feeding method with 42 selection episodes over 45 generations. The cotton bollworm developed resistance to transgenic Bt cotton after 12 generations (F12) of selection. The survival rate of F12 neonates feeding on leaves of seedling stage for 4 days and boll-opening stage for 5 days of R19 line were ca 34 and 72%, respectively, compared with ca 0 and 40% for a non-selected sister strain (NYCS), but lower than or similar to that of F12 feeding on leaves of non-Bt cotton, Sumian 12 (ca 87 or 76%). Resistance to B thuringiensis HD-1 Dipel in neonates (F12) was ca 6-fold. After 42 generations of selection, the strain developed a very high level of resistance to Cry1Ac protoxin, 210 g kg(-1) MVPII wettable powder and 200 g litre(-1) MVPII liquid formulation, the resistance ratios being 1680-, 1780- and ca 1200-fold, respectively, compared with a laboratory susceptible strain (HZS). When compared with the non-selected NYCS, the resistance ratios to the above toxins were ca 540-, 580- and 510-fold, respectively, but to Dipel only ca 16-fold. The results indicated that it is very important to develop and implement effective resistance-management strategies and to detect early resistance to Bt cotton in field populations.     

郭予元院士在棉铃虫区域性综合防治技术体系研究及应用中的学术贡献——纪念郭予元院士诞辰90周年

郭予元院士带领团队研究人员通过多年协同攻关研究，解析了棉铃虫发生为害规律和自然种群生命表机制，明确了棉铃虫对多种农药抗药性动态并应用于棉田抗药性治理，揭示了黄河流域棉区棉花对不同世代棉铃虫为害的补偿生长规律和防治策略，基于不同世代棉铃虫为害阈值和防治指标创建了准确预报二代棉铃虫发生数量的一代麦田扫网法，开展棉花种质资源抗病虫性鉴定并指导选育多抗棉花系列新品种。通过整合诱杀、选择性杀虫剂、生物农药与化学农药次序使用和局部精准施药等绿色防治技术，组建区域性棉花主要病虫害综合防治技术体系，棉区棉铃虫为害得到有效控制，引领了我国农作物主要病虫害综合防治技术的发展。本文简要回顾了郭予元院士在该领域研究中做出的主要历史贡献。     

抗Bt棉铃虫种群的交互抗性研究

分别用对CyrlAc抗性达11．6倍的Bt棉叶和Bt粉剂（Btk)汰选棉铃虫（Heliocoverpa armigera)种群（对Btk的抗性指数为5．2倍）,测定了其对不同Bt制剂,Bt毒素,化学农药的交互抗性,结果表明：停止汰选后,Bt棉叶汰选种群对CrylAc的抗性指数为5．9倍,对CrylAc 1C的交互抗性为3．7倍,对Btk的敏感性降低,对Bta,Cry2A无交互抗性,Btk汰选种群对Btk的抗性指数为4．1倍,对Bta,CrylAc,CrylAc 1C的交互抗性为2．3,3．1,2．3倍,对Cry2A无交互抗性,两汰选种群对化学农药灭多威,辛硫磷,氯氰菊酯的敏感性均有所增加。     

国外草地贪夜蛾化学防治技术的研究与应用

草地贪夜蛾Spodoptera frugiperda(J. E. Smith)是一种适应性和迁徙能力很强的农业害虫,并具有暴发为害的特点。几十年来,化学防治一直是防治草地贪夜蛾的重要技术措施。20世纪80年代以前,美洲国家防治草地贪夜蛾以有机磷类和氨基甲酸酯类杀虫剂为主;随后拟除虫菊酯类杀虫剂得到了广泛使用;20世纪90年代中后期,美洲特别是美国推广种植转基因Bt玉米后,防治草地贪夜蛾杀虫剂的用量显著减少;近年来,甲氨基阿维菌素苯甲酸盐、乙基多杀菌素、除虫脲、虱螨脲、茚虫威、氯虫苯甲酰胺、氟苯虫酰胺、溴氰虫酰胺等一系列新型杀虫剂品种在防治草地贪夜蛾中发挥了突出的作用。伴随着不同种类杀虫剂的使用,草地贪夜蛾对杀虫剂抗性也相应地呈现出明显的变化,20世纪80年代后,美洲的草地贪夜蛾先后对有机磷类和氨基甲酸酯类杀虫剂产生了抗药性,截至2017年,美洲地区的草地贪夜蛾至少对包括氨基甲酸酯类、有机磷类、拟除虫菊酯类及Bt杀虫蛋白等不同类型共29种杀虫剂产生了抗药性;2016年,采自北美洲波多黎各的草地贪夜蛾田间种群对双酰胺类杀虫剂氟苯虫酰胺和氯虫苯甲酰胺抗性倍数最高分别达500倍和160倍。草地贪夜蛾的抗药性机制主要是解毒代谢增强和靶标基因突变。种子处理可以预防作物苗期草地贪夜蛾的为害,并且有利于天敌保护。雾滴粒径不仅影响杀虫剂对草地贪夜蛾的防治效果,还影响草地贪夜蛾对杀虫剂抗药性的发展,因此,防治田间草地贪夜蛾适宜采用细雾喷雾方式。使用得当,化学杀虫剂可以与天敌和生物杀虫剂协同增效防治草地贪夜蛾。航空施药要和地面施药联合使用才能对玉米田严重发生的草地贪夜蛾达到好的防治效果。     

Does cotton bollworm show cross-resistance to the Bacillus thuringiensis toxins Cry1Ac and Cry2Ab? A mini review

Since 1996, transgenic Bacillus thuringiensis(Bt) cotton has been commercially grown in numerous countries in an effort to stem the losses caused by key lepidopteran pests. However, the development of pest resistance to Bt toxins has jeopardized the continued utilization of Bt cotton. As a strategy designed to circumvent the development of resistance, Bt cotton varieties expressing two or more toxins targeting the same pest have been introduced. Nevertheless, from the perspective of long-term planting of Bt cotton, the potential risk of cross-resistance to these Bt toxins is a threat that cannot be ignored. In this paper, we review current research(including that based on the analysis of protein binding sites and resistance genes) on the resistance of cotton bollworm(Helicoverpa armigera) to the Bt toxins Cry1 Ac and Cry2 Ab and the interrelationship between these toxins. On the basis of existing evidence, we assume that the actions of Cry1 Ac and Cry2 Ab against cotton bollworm are not completely independent, and then propose the "resistance-associated gene mutation potential hypothesis". Although the mechanisms underlying the resistance of pests to Bt toxins are yet to be comprehensively elucidated, this hypothesis could undoubtedly have important implications for adopting "pyramid" strategy in the future. Further research is recommended to devise strategies to retard the development of H. armigera resistance to Bt cotton, either using different Bt toxins or their various combinations.     

新疆南部转基因棉区棉铃虫种群长期动态研究

（Bacillus thuringiensis）基因抗虫棉在新疆南部推广已超过10 a,为探索Bt棉大面积种植对棉铃虫（Helicoverpa armigera）种群动态的影响,于1999-2010年使用诱虫灯监测了Bt棉大面积种植区域（麦盖提）和非Bt棉大面积种植区域（阿瓦提）棉铃虫的种群动态。结果表明：新疆南部地区20世纪90年代末发生棉铃虫害较重,2000-2004年种群数量保持较高水平;自2005年Bt棉大面积推广以后,Bt棉区棉铃虫的种群数量显著下降,棉铃虫种群数量随Bt棉种植比例的上升而下降（P<0.05）;]随着Bt棉大面积推广年数的增加,Bt棉区棉铃虫各代种群数量均逐渐下降,第二代种群数量和高峰期蛾量下降速率均较越冬代和第一代快,且第二代棉铃虫种群相对丰富度也逐渐下降。因此,新疆地区Bt棉的大面积种植能较好地控制棉铃虫的种群数量,而且对第二代棉铃虫种群的控制效果最好。     

A perspective on management of Helicoverpa armigera: transgenic Bt cotton,IPM, and landscapes

Helicoverpa armigera is a major pest of agriculture, horticulture and floriculture throughout the Old World and recently invaded parts of the New World. We overview of the evolution in thinking about the application of area‐wide approaches to assist with its control by the Australian Cotton Industry to highlight important lessons and future challenges to achieving the same in the New World. An over‐reliance of broad‐spectrum insecticides led to Helicoverpa spp. in Australian cotton rapidly became resistant to DDT, synthetic pyrethroids, organophosphates, carbamates and endosulfan. Voluntary strategies were developed to slow the development of insecticide resistance, which included rotating chemistries and basing spray decisions on thresholds. Despite adoption of these practices, insecticide resistance continued to develop until the introduction of genetically modified cotton provided a platform for augmenting Integrated Pest Management in the Australian cotton industry. Compliance with mandatory resistance management plans for Bt cotton necessitated a shift from pest control at the level of individual fields or farms towards a coordinated area‐wide landscape approach. Our take‐home message for control of H. armigera is that resistance management is essential in genetically modified crops and must be season long and area‐wide to be effective. © 2016 Society of Chemical Industry     

V-ATPase H表达量下调增强棉铃虫对Cry1Ac的抗性

棉铃虫Helicoverpa armigera是世界性重要农业害虫。目前防治棉铃虫的主要手段是种植转苏云金芽胞杆菌Bacillus thuringiensis(Bt)杀虫蛋白的转基因作物。本文旨在研究棉铃虫V-ATPase H在Cry1Ac蛋白毒力和抗性中的作用。利用实时荧光定量qRT-PCR技术分析V-ATPase H在Cry1Ac抗、感品系棉铃虫幼虫中肠及敏感品系棉铃虫幼虫受Cry1Ac诱导后的表达情况;在昆虫Sf9细胞中过表达V-ATPase H对其进行细胞定位,通过细胞毒力试验验证其对Cry1Ac毒力的影响。结果发现棉铃虫V-ATPase H基因在抗性品系中低表达,并且V-ATPase H在受到Cry1Ac诱导时也低表达;在Sf9细胞内表达V-ATPase H蛋白发现其在整个细胞中都有分布,过表达该蛋白后增强了细胞对Cry1Ac蛋白的敏感性。结果表明V-ATPase H参与Cry1Ac蛋白的毒力。     

Toxicity of chlorantraniliprole to Cry1Ac-susceptible and resistant strains of Helicoverpa armigera

Transgenic Bt cotton expressing Cry1Ac is important in controlling various agricultural pests, including Helicoverpa armigera. Especially for transgenic crops that are cultivated in large expanses, avoiding resistance development is a key for ensuring sustainability of Bt technologies. Integrated pest management, in which transgenic crops are strategically combined with rational pesticide use, may help to prevent H. armigera resistance acquisition in Bt cotton. In this study, we evaluated the toxicity of a novel insecticide (chlorantraniliprole) on Cry1Ac-susceptible and resistant individuals of H. armigera. More specifically, we assessed the effect of chlorantraniliprole on the activity of two enzymes and conducted laboratory bioassays to determine its toxicity on H. armigera larvae. Chlorantraniliprole increased esterase and glutathione-S-transferase activities in Cry1Ac susceptible and resistant populations of H. armigera. Cry1Ac resistant populations XJ-F (Cry1Ac resistance ratio 21.8-fold), XJ-10.0 (95.8-fold) and BTR (3536.5-fold) did not show cross-resistance to chlorantraniliprole, with LC₅₀ values of 0.0733 (μg/mL) in XJ-F, 0.0545 (μg/ml) in XJ-10.0 and 0.0731 (μg/mL) in BTR, which were close to that in the susceptible strain 96S (0.0954 μg/mL). Our work shows that chlorantraniliprole could be considered to be integrated in Bt cotton management schemes to delay the H. armigera resistance development.     

草地贪夜蛾对化学农药和Bt作物的抗性机制及其治理技术研究进展

草地贪夜蛾Spodoptera frugiperda(Smith)是一种重要的经济作物害虫,在美洲玉米种植区均有发生。近几年该害虫迅速蔓延至非洲和东南亚等地区,并于2018年12月下旬在中国云南省被发现,对玉米生产造成严重威胁。化学农药和Bt作物是目前防治草地贪夜蛾的最主要手段,但大量研究表明,草地贪夜蛾已经形成了具有高抗药性和Bt抗性的种群。本文介绍了草地贪夜蛾抗药性和Bt抗性现状,从抗性相关因子的表达调控和靶标位点变异2个方面论述了草地贪夜蛾抗药性和Bt抗性机制,总结了目前草地贪夜蛾抗药性和Bt抗性的治理策略,并结合中国实际情况探讨了草地贪夜蛾综合防控的发展方向,以期为该害虫的防控提供参考。     

Biochemical and physiological changes on Bacillus thuringiensis cotton after imidacloprid foliar spray

Bacillus thuringiensis cotton is a variety of cotton genetically modified to contain a gene derived from B. thuringiensis bacteria; which results in expression of toxin protein that confers resistance to bollworm complex (the most destructive pest of cotton). Introduction of Bt cotton lowered the need of insecticides, but still a number of insecticides are used for other insects like jassids, whitefly, aphids and tobacco caterpillar to which Bt gene does not provide effective control. Imidacloprid (tradename Imidacel 17.8 SL) is an insecticide designed for control of these major sucking/piercing insects that affect cotton. So in the present work we studied the post effect of imidacloprid insecticide on plant health of three Bt cotton hybrids (RCH-134, JKCH-1947, NCEH-6R) as there are reports of this insecticide causing growth and yield enhancements in absence of insect pests. Imidacloprid was first sprayed at recommended concentration (40 ml/acre) on 3 months old plants sown in randomly designed plots with three replications of each hybrid. The spray was repeated three times at 10 days interval. The level of B. thuringiensis gene expression, peroxidase activity and total phenols was measured on third day after every spray in leaves along with growth and yield of plants. The insecticide has shown to increase the level of B. thuringiensis protein, peroxidase enzyme activity, total phenols, height, number of bolls retained on plants and yield. These observations suggested that the imidacloprid treated plants showed better growth and development, thereby imidacloprid has growth enhancing effect on Bt cotton plants in addition to its insecticidal properties.     

转Bt基因棉在安庆市大面积种植后害虫发生特点调查

根据调查,转Bt基因棉大面积种植后,棉铃虫、红铃虫发生减轻;棉叶螨发生加重,为害时间延长;甜菜夜蛾、斜纹夜蛾发生面积扩大,为害程度加重。据此初步提出了当地转Bt基因棉害虫防治中应注意的问题和管理对策。     

红铃虫对Bt棉花抗性治理的研究进展

综述了Bt棉花对红铃虫的抗性效率和红铃虫对Bt棉花的抗性机理、抗性遗传、抗性监测及治理对策的研究进展.研究结果表明,不同种植区Bt棉花对红铃虫的抗性水平有较大差别.红铃虫中肠内Bt毒素结合位点数量减少是产生抗性的主要机制.抗性由具1个主效应的多位点基因控制,高水平抗性的产生与编码钙蛋白基因变异有关.避难所策略是红铃虫抗性治理的主要措施.抗性监测表明,自1997年以来,美国亚历桑那州红铃虫田间种群对Bt棉花的抗性频率无显著性变化.