甲氰菊酯与阿维菌素单用、轮用和混用对朱砂叶螨抗性进化的影响

应用数量遗传学的方法分析朱砂叶螨(Tetranychus cinnabarinus)实验种群对甲氰菊酯、阿维菌素及其混剂甲氰—阿维(甲氰菊酯:阿维菌素=8.9:0.1,m/m)的抗性现实遗传力,并测定了甲氰菊酯、阿维菌素分别连续单用、轮换使用、混合使用对朱砂叶螨抗性进化的影响。结果表明,筛选16代后,朱砂叶螨对甲氰菊酯、阿维菌素和甲氰—阿维的抗性现实遗传力分别为0.2853、0.1695和0.0804,朱砂叶螨对混剂的抗性现实遗传力低于对2个单剂的遗传力的一半,混用延缓抗性的效果将好于轮用。药剂连续单用、轮换使用和混合使用16代,朱砂叶螨对甲氰菊酯的抗性分别为28.52、28.03和10.81倍,对阿维菌素的抗性分别为3.24、2.82和1.41倍。朱砂叶螨对2种杀螨剂抗性进化速率为单用>轮用>混用,抗性测定结果表明甲氰菊酯与阿维菌素混用能有效延缓朱砂叶螨对2种药剂抗性的发展速率。     

二斑叶螨田间种群对阿维菌素的抗性及抗性相关基因表达分析

【目的】二斑叶螨Tetranychus urticae Koch是为害多种农作物的世界性重大害螨。本研究旨在明确二斑叶螨不同田间种群对阿维菌素的抗药性、抗性相关基因的突变频率及其表达量变化。【方法】采用药管浸叶法测定了我国二斑叶螨8个地理种群对阿维菌素的抗药性并检测其抗性基因突变频率,结合荧光定量PCR技术分析了高抗种群中抗性相关基因表达量变化。【结果】测试的二斑叶螨8个田间种群均对阿维菌素具有抗药性。北京密云、山东潍坊、海南三亚和湖南长沙种群均对阿维菌素产生了极高水平的抗性,抗性倍数分别为1 526.75,481.00,315.25和160.75倍,而北京通县、北京海淀、山西运城和山东泰安种群对阿维菌素的抗性倍数达54.38~136.38倍,处于高抗性水平。二斑叶螨对阿维菌素抗性相关的谷氨酸氯离子通道基因Glu Cl的突变频率在各个田间种群中存在差异。北京密云种群中Glu Cl的突变频率最高(91.7%),其次是山东潍坊(66.7%)和海南三亚(63.3%)种群;山西运城种群最低(13.3%),且点突变频率与抗性倍数之间呈显著的正相关(P0.05)。与相对敏感种群相比,高抗性二斑叶螨种群中Glu Cl和γ-氨基丁酸受体基因GABAR表达量显著下降。【结论】二斑叶螨田间种群普遍对阿维菌素产生了高水平抗性,抗性相关基因Glu Cl的点突变及其表达量的降低可能与田间抗药性产生相关;田间防治二斑叶螨应避免使用阿维菌素。     

朱砂叶螨对甲氰菊酯、阿维菌素,哒螨灵及其混剂抗性遗传力的分析(英文)

采用域性状分析法 ,估算了朱砂叶螨对 5种杀螨剂 (3种单剂和 2种混剂 )的抗性现实遗传力 ,并对 5种药剂的抗性风险进行了评估。把采自重庆北碚田间的朱砂叶螨种群 ,在室内不施药情况下饲养 6 0余代 ,以此作为抗性筛选的敏感品系。分别单一连续汰选近 30代 ,朱砂叶螨对甲氰菊酯、阿维菌素、哒螨灵、哒螨 -阿维 (哒螨灵 :阿维菌素 =7 4 :0 1,m m)和甲氰 -阿维 (甲氰菊酯 :阿维菌素 =8 9:0 1,m m)的抗性分别达 6 5 5 5、5 82、1 2 3、5 2 0和 1 4 2 倍 ;抗性现实遗传力分别为 0 2 16 7、0 0 96 7、0 0 130、0 0 80 0和 0 0 172。在实验室选择条件下 ,预计抗性增长 10倍时 ,甲氰菊酯、阿维菌素、哒螨灵、哒螨 -阿维 (哒螨灵 :阿维菌素 =7 4 :0 1,m m)和甲氰 -阿维 (甲氰菊酯 :阿维菌素 =8 9:0 1,m m)分别需要 15、34、333、4 2和 2 0 0代。甲氰菊酯抗性风险较高 ,其次是阿维菌素、哒螨 -阿维 (哒螨灵 :阿维菌素 =7 4 :0 1,m m)、甲氰 阿维 (甲氰菊酯 :阿维菌素 =8 9:0 1,m m) ,哒螨灵抗性风险较低。混剂哒螨 阿维 (哒螨灵 :阿维菌素 =7 4 :0 1,m m)不能延缓朱砂叶螨对两单剂哒螨灵和阿维菌素的抗性发展 ,而混剂甲氰 阿维 (甲氰菊酯 :阿维菌素 =8 9:0 1,m m)却能有效延缓朱砂叶螨对两单剂     

土耳其斯坦叶螨多重抗性品系选育及解毒酶活性的变化

为探索土耳其斯坦叶螨的多重抗药性及其生化机理,在室内对敏感系(SS)土耳其斯坦叶螨分别用螺螨酯、甲氰菊酯和阿维菌素的混剂进行处理,选育出多重抗性品系(Mp-R).结果表明: 选育至15代,土耳其斯坦叶螨的抗性指数达35.74倍.对不同品系的解毒酶活性分析显示,Mp-R品系相对SS品系的羧酸酯酶(CarE)、谷胱甘肽-S-转移酶(GSTs)和多功能氧化酶(MFO)的比活力分别是SS品系的1.21、1.53、9.18倍.说明CarE、GSTs、MFO的活性升高可促进土耳其斯坦叶螨对3种杀虫剂多重抗性的形成;MFO的活性升高可能是土耳其斯坦叶螨对3种杀虫剂产生多重抗性的主要原因.测定Mp-R品系和单抗品系(Ip-R)的农药感性和解毒酶活力变化发现,3种杀虫剂的混合使用可能会延缓土耳其斯坦叶螨对甲氰菊酯的抗性形成,加快对阿维菌素的抗性形成.     

朱砂叶螨对三种杀螨剂的抗性选育与抗性风险评估

为评价朱砂叶螨Tetranychus cinnabarinus对3种杀螨剂的抗性风险,在实验室抗性品系选育基础上,应用数量遗传学中的域性状分析法,研究了朱砂叶螨北碚种群对甲氰菊酯、阿维菌素和哒螨灵3种杀螨剂的抗性现实遗传力,并对3种药剂在不同杀死率下抗性发展的速率进行了预测。结果表明：分别单一连续汰选16代后,朱砂叶螨对甲氰菊酯、阿维菌素的抗性倍数分别达26.54和4.51倍,对哒螨灵表现为敏感性降低（抗性倍数为1.16倍）;朱砂叶螨对甲氰菊酯、阿维菌素和哒螨灵的抗性现实遗传力分别为0.2472,0.1519和0.0160。在室内选择条件下,杀死率为50%～90%时,要获得10倍抗性,甲氰菊酯仅需要13～6代,阿维菌素需要约21～10代;哒螨灵需要约197～89代;在田间选择,三种药剂都将需要更长的时间。抗性筛选16代结果表明,抗性风险较高的是菊酯类的甲氰菊酯,其次是生物源农药阿维菌素,杂环类的哒螨灵抗性风险较小。试验结果可为朱砂叶螨抗性治理提供参考。     

二斑叶螨对阿维菌素、哒螨灵和甲氰菊酯的抗性选育及其解毒酶活力变化

在室内模拟田间药剂的选择压力,用阿维菌素、哒螨灵和甲氰菊酯对二斑叶螨Tetranychuc urticae逐代处理,以选育其抗性种群。选育至12代,对阿维菌素抗性增长到6.72倍,对哒螨灵抗性增长到12.1倍,对甲氰菊酯抗性增长到19.9倍。酶抑制剂和离体酶活性的测定结果表明,阿维菌素抗性种群的多功能氧化酶和谷胱甘肽S-转移酶的活性均有所提高;二斑叶螨对哒螨灵的抗性可能与多功能氧化酶、羧酸酯酶的活性增强有关;而羧酸酯酶、多功能氧化酶和谷胱甘肽S-转移酶活性的增强可能是二斑叶螨对甲氰菊酯产生抗性的主要原因。     

朱砂叶螨对甲氨菊酯、阿维菌素，哒螨灵及其混剂抗性遗传力的分析

采用域性状分析法,估算了朱砂叶螨对5种杀螨剂(3种单剂和2种混剂)的抗性现实遗传力,并对5种药剂的抗性风险进行了评估。把采自重庆北碚田间的朱砂叶螨种群,在室内不施药情况下饲养60余代,以此作为抗性筛选的敏感品系。分别单一连续汰选近30代,朱砂叶螨对甲氰菊酯、阿维菌素、哒螨灵、哒螨—阿维(哒螨灵：阿维菌素＝7．4：0．1,m/m)和甲氰—阿维(甲氰菊酯：阿维菌素＝8．9：0．1,m/m)的抗性分别达65．55、5．82、1．23、5．20和1．42．倍;抗性现实遗传力分别为0．2167、0．0967、0．0130、0．0800和0．0172。在实验室选择条件下,预计抗性增长10倍时,甲氰菊酯、阿维菌素、哒螨灵、哒螨—阿维(哒螨灵：阿维菌素＝7．4：0．1,m/m)和甲氰—阿维(甲氰菊酯：阿维菌素＝8．9：0．1,m/m)分别需要15、34、333、42和200代。甲氰菊酯抗性风险较高,其余是阿维菌素、哒螨—阿维(哒螨灵：阿维菌素＝7．4：0．1,m/m)、甲氰-阿维(甲氰菊酯：阿维菌素＝8．9：0．1,m/m),哒螨灵抗性风险较低。混剂哒螨-阿维(哒螨灵：阿维菌素＝7．4：0．1,m/m)不能延缓朱砂叶螨对两单剂哒螨灵和阿维菌素的抗性发展,而混剂甲氰-阿维(甲氰菊酯：阿维菌素＝8．9：0．1,m/m)却能有效延缓朱砂叶螨对两单剂甲氰菊酯和阿维菌素的抗性发展。     

截形叶螨对哒螨灵、阿维菌素和阿维·哒螨灵的抗性选育和抗性稳定性研究

【目的】明确截形叶螨Tetranychus truncatus Ehara对哒螨灵、阿维菌素和阿维·哒螨灵3种田间常用药剂产生抗性的速率和稳定性,为叶螨的抗性综合治理提供一定的理论依据。【方法】采用室内生测法,对截形叶螨进行药剂的抗性筛选、衰退和再恢复规律研究。【结果】经过连续30代的药剂汰选,截形叶螨对哒螨灵、阿维菌素和阿维·哒螨灵3种药剂产生了不同程度的抗药性,抗性指数分别达到197.50、19.56和12.57;停止喷药后,其抗性都有所下降,其中截形叶螨对哒螨灵的抗性最不稳定,培育至30代后,抗性衰退率达到63.54%,对阿维菌素的抗性较为稳定,抗性衰退率为23.30%;再次恢复用药后,截形叶螨对哒螨灵、阿维菌素和阿维哒螨灵抗性再度回升,以抗哒螨灵品系的抗性恢复最快,药剂汰选30代后,增长率达到了58.47%,阿维·哒螨灵次之(增长率为38.67%),抗阿维菌素的品系抗性恢复最慢,增长率仅为22.86%。【结论】截形叶螨对哒螨灵抗性不稳定,停止用药后,敏感性易恢复,对阿维菌素和阿维·哒螨灵的抗性较稳定,一旦抗性产生不易衰退,故田间应用时应交替轮换用药。     

桔全爪螨对阿维菌素和甲氰菊酯的抗性现实遗传力及风险评估

在实验室抗性选育的基础上,应用数量遗传学中的域性状分析法,研究了桔全爪螨北碚种群对阿维菌素和甲氰菊酯2种杀螨剂的抗性现实遗传力,并对2种药剂在不同杀死率下抗性发展的速率进行了预测.结果表明:用阿维菌素和甲氰菊酯分别不连续汰选11及16代后,桔全爪螨对两者的抗性分别为3.8和29.9倍,抗性现实遗传力分别为0.0475和0.1544.在室内选择条件下,杀死率为50%～90%时,要获得10倍抗性,甲氰菊酯仅需要7～16代,阿维菌素则需要12～26代.而在田间选择情况下,2种药剂都将需要更长的时间.抗性筛选结果表明,生物源农药阿维菌素的抗性风险明显低于菊酯类农药甲氰菊酯.试验结果可为桔全爪螨抗性治理提供参考.     

智利小植绥螨饲养释放试验

<正> 柑桔全爪螨Panonychus citri(McGregor)是柑桔上的主要害虫之一。第二次世界大战后,由于长期使用有机磷等农药,引起了叶螨的猖獗为害,且其残毒及害虫的抗性已成为难以解决的问题。现在,害螨已成为棉花、小麦、果树及温室蔬菜上的大害虫。利用益螨防治害螨近年来的研究进展很快,引人瞩目的首推智利小植绥螨Phytosciulus persimilis Athias-Henriot,目前,北美、欧洲许多国家均已商品化,普遍用于防治温室内的叶螨及温室外棉花、土豆、瓜类、苹果、     

Insecticides with novel modes of action: Mechanism, selectivity and cross-resistance

Efforts have been made during the past two decades to develop insecticides with selective properties that act specifically on biochemical sites present in particular insect groups, but whose properties differ from other insecticides. This approach has led to the discovery of compounds that affect the hormonal regulation of molting and developmental processes in insects; for example, ecdysone agonists, juvenile hormone mimics and chitin synthesis inhibitors. In addition, compounds that selectively interact with the insect nicotinic acetylcholine receptor, such as imidacloprid, acetamiprid and thiamethoxam, have been introduced for the control of aphids, whiteflies and other insect species. Natural products acting selectively on insect pests, such as avermectins, spinosad and azadirachtin, have been introduced for controlling selected groups of insect pests. Compounds acting on the nervous site that controls the sucking pump of aphids and whiteflies, such as pymetrozine, or respiration, such as diafenthiuron, have been introduced for controlling sucking pests. All the above compounds are important components in pest and resistance management programs.     

Herbicide resistance—what have we learned from other disciplines?

Herbicide resistance is a growing threat to agriculture and has parallels to resistances to fungicides and insecticides. However, there are many reasons to treat the resistance to herbicides differently. To highlight these similarities and differences, three pests, a weed, an insect, and a disease that have shown the ability to rapidly develop resistance to a variety of products and product classes were used as illustrations. The situation in herbicide resistance is approaching a point already experienced by the other pest control disciplines, and thus, it is worthwhile to revisit their experiences.     

钠离子通道与蜜蜂狄斯瓦螨对氟胺氰菊酯的抗性机理

狄斯瓦螨Varroadestructor是全世界蜜蜂最严重的寄生虫 ,目前 ,它对主要防治药物———拟除虫菊酯类的氟胺氰菊酯已产生明显抗性 ,严重影响其防治效果。近年来神经生理学研究结果证实 :电压门控的钠离子通道是拟除虫菊酯作用的位点。钠通道结构的改变 ,是拟除虫菊酯类杀虫剂毒理的主要基础 ,也是产生抗药性的基础。该文介绍了近年来国内外研究电压门控钠离子通道、拟除虫菊酯对钠通道的作用、钠通道与拟除虫菊酯的抗性和狄斯瓦螨对氟胺氰菊酯抗性机理研究的新进展     

Detection and monitoring of insect resistance to transgenic Bt crops

Transgenic crops expressing Bacillus thuringiensis （Bt） endotoxins havebecome one of the most important tools for managing corn and cotton insect pests in the US and other countries. The widespread adoption of transgenic Bt crops could place a high degree of selection pressure on the target insect populations and accelerate development of resistance, raising concerns about the long-term durability of Bt plants as an effective pest management tool. Conservation of Bt susceptibility in insects has become one of the most active research areas in modern agriculture. One of the key factors for a successful Bt resistance management plan is to have a cost-effective monitoring system that can provide information on. （i） the initial Bt resistance allele frequencies at low levels in field insect populations; and （ii） early shifts in Bt resistance allele frequencies so that proactive measures for managing resistance can be deployed well before field control failures. Developing such a monitoring program has been difficult because： （i） resistance traits that occur at very low frequencies are hard to detect; （ii） many factors affect the sensitivity and accuracy of a Bt resistance monitoring program; and （iii） monitoring resistance is costly. Several novel methods for detecting Bt resistance alleles developed during the last decade have made a cost-effective monitoring system possible. Future studies should focus on how to improve and standardize the methodologies for insect sampling and Bt resistance detection.     

The progress in insect cross‐resistance among Bacillus thuringiensis toxins

Bt crop pyramids produce two or more Bt proteins active to broaden the spectrum of action and to delay the development of resistance in exposed insect populations. The cross‐resistance between Bt toxins is a vital restriction factor for Bt crop pyramids, which may reduce the effect of pyramid strategy. In this review, the status of the cross‐resistance among more than 20 Bt toxins that are most commonly used against 13 insect pests was analyzed. The potential mechanisms of cross‐resistance are discussed. The corresponding measures, including pyramid RNA interference and Bt toxin, “high dose/refuge,” and so on are advised to be taken for adopting the pyramided strategy to delay the Bt evolution of resistance and control the target pest insect.     

昆虫抗药性产生机制

杀虫剂是害虫防治的有效途径之一,但随着杀虫剂长期和广泛的使用,昆虫种群对各种杀虫剂的敏感性降低,产生了抗药性,如何克服昆虫的抗药性是害虫综合治理的重要问题。近年来,借助基因组测序和遗传操作技术的发展,对昆虫抗药性的研究已经深入到细胞水平和分子水平,取得诸多重要的突破,为害虫抗性的控制奠定了理论基础。本文从常见杀虫剂的历史沿革及作用机理切入,从靶标抗性、代谢抗性和穿透抗性3个方面阐述了杀虫剂抗性产生的机制:杀虫剂作用位点的突变降低了靶标与杀虫剂的亲和力,细胞色素P450酶系和谷胱甘肽转移酶系的激活增加了杀虫剂的降解,表皮结构成分的变化和ABC转运蛋白的增加有效阻挡了杀虫剂的渗入。利用基因操作手段或抑制剂,对上述3种抗性机制的关键步骤进行调控可能成为未来杀虫剂抗性控制的新策略。     

Genome mapping coupled with CRISPR gene editing reveals a P450 gene confers avermectin resistance in the beet armyworm

The evolution of insecticide resistance represents a global constraint to agricultural production. Because of the extreme genetic diversity found in insects and the large numbers of genes involved in insecticide detoxification, better tools are needed to quickly identify and validate the involvement of putative resistance genes for improved monitoring, management, and countering of field-evolved insecticide resistance. The avermectins, emamectin benzoate (EB) and abamectin are relatively new pesticides with reduced environmental risk that target a wide number of insect pests, including the beet armyworm, Spodoptera exigua, an important global pest of many crops. Unfortunately, field resistance to avermectins recently evolved in the beet armyworm, threatening the sustainable use of this class of insecticides. Here, we report a high-quality chromosome-level assembly of the beet armyworm genome and use bulked segregant analysis (BSA) to identify the locus of avermectin resistance, which mapped on 15–16 Mbp of chromosome 17. Knockout of the CYP9A186 gene that maps within this region by CRISPR/Cas9 gene editing fully restored EB susceptibility, implicating this gene in avermectin resistance. Heterologous expression and in vitro functional assays further confirm that a natural substitution (F116V) found in the substrate recognition site 1 (SRS1) of the CYP9A186 protein results in enhanced metabolism of EB and abamectin. Hence, the combined approach of coupling gene editing with BSA allows for the rapid identification of metabolic resistance genes responsible for insecticide resistance, which is critical for effective monitoring and adaptive management of insecticide resistance.     

转Bt基因抗虫棉的生态风险及治理对策

评述了转Bt基因抗虫棉的生态风险及治理对策。其生态风险主要表现在目标害虫的抗性和对非目标生物群落的变化。目标害虫与转基因抗虫棉的互相作用和抗虫棉杀虫毒素的时空表达方式是目标害虫抗性发展的主要途径。在转基因抗虫棉田中,虽然对目标害虫的防治次数大为减少,但害虫和天敌群落的稳定性仍不如常规棉田,某种次要害虫大发生的可能性较大。认为将转基因抗虫棉纳入综合防治体系并培育更加高效的抗虫棉是治理目标害虫抗性和防止次要害虫上升的重要措施。     

北京和湖南烟粉虱生物型及其抗药性监测

烟粉虱是蔬菜等园艺作物上为害严重、最难以防治的害虫之一,生产上防治该虫主要还是依靠化学药剂。本研究鉴定了北京、湖南地区设施蔬菜上烟粉虱的生物型,并监测了其对6种不同杀虫剂的抗药性。结果表明,北京、湖南地区蔬菜烟粉虱生物型分别是Q型和B型。两地烟粉虱对阿维菌素尚处于敏感状态;对烯啶虫胺及其他烟碱类杀虫剂噻虫嗪和吡虫啉的抗药性达中抗到高抗水平,湖南地区抗性水平比北京地区更高,抗性倍数最高达到71.58倍;传统杀虫剂毒死蜱和联苯菊酯对湖南、北京两地烟粉虱的毒杀活性差。     

Conserving the efficacy of insecticides against Plutella xylostella (L.) (Lep., Plutellidae)

Abstract:  The diamondback moth (DBM), Plutella xylostella (L.) (Lep., Plutellidae), is one of the most destructive insect pests of crucifers worldwide. It was the first crop insect reported to be resistant to DDT and now in many crucifer-producing regions it has shown significant resistance to almost every insecticide applied in field including biopesticides such as crystal toxins from Bacillus thuringiensis and spinosyns from Saccharopolyspora spinosa . In certain parts of the world, economical production of crucifers has become almost impossible because of its resistance to insecticides and resulting control failure. A coordinated resistance management program needs to be implemented with the involvement of pesticide industry, local pesticide regulatory authorities, scientists and farmers. The judicious use of chemicals in conjunction with other control measures (e.g. biological control agents, resistant varieties, proper fertilization rates) is the best way to manage DBM and other pests of cruciferous crops. Introduction of glucosinolate-sulphatase inhibitors as plant-incorporated-products or sprayable material may also lead to a novel pest management strategy.