Effect of Melia azedarach extract on the activity of NADPH-cytochrome c reductase and cholinesterase in insects

The effect of Melia azedarach extract on the activity of NADPH-cytochrome c reductase and cholinesterase was studied in Spodoptera frugiperda. Larvae were fed an artificial diet containing fruit extract and their midgut was used for enzyme determination. As compared to the control, consumption of the extract containing diet resulted in a 31% inhibition of the cholinesterase activity and a 34% activation of the NADPH-cytochrome c reductase activity. In the cockroach Leucophaea maderae, the effect on reductase activity was even more pronounced (43%).     

菊酯类药剂对草地贪夜蛾的毒力及对钠离子通道的诱导特性

为明确菊酯类药剂对草地贪夜蛾Spodoptera frugiperda的防治效果,采用点滴法在室内测定高效氯氟氰菊酯、高效氯氰菊酯、联苯菊酯、溴氰菊酯和茚虫威原药对草地贪夜蛾3龄幼虫的毒力,利用实时荧光定量PCR技术测定3龄幼虫在这5种药剂诱导不同时间下其体内钠离子通道基因片段的表达量变化。结果显示,高效氯氟氰菊酯、溴氰菊酯、联苯菊酯、茚虫威和高效氯氰菊酯这5种药剂处理草地贪夜蛾3龄幼虫24 h后的LD50值分别为74.911、83.280、152.662、179.372、567.250 mg/L;处理48 h后的LD50值分别为18.946、67.874、120.888、132.790、461.635 mg/L;同时发现草地贪夜蛾3龄幼虫钠离子通道基因的表达均呈现不被诱导或诱导下调现象,表明草地贪夜蛾幼虫可通过降低靶标基因的表达量进而减少药剂对其的毒性。表明供试4种拟除虫菊酯类药剂和1种氯虫苯甲酰胺类药剂可作为草地贪夜蛾田间应急防控药剂施用。     

Fitness cost, cross resistance and realized heritability of resistance to imidacloprid in Spodoptera litura (Lepidoptera: Noctuidae)

Imidacloprid has been used as a key insecticide for controlling sucking insect pests of cotton, whereas Spodoptera litura also has been indirectly exposed to this insecticide in Pakistan. To evaluate the risk of resistance evolution and to develop a better resistance management strategy, a field collected population was selected with imidacloprid in the laboratory. Thereafter, fitness cost, realized heritability and cross resistance of imidacloprid resistance in S. litura were investigated. After 14 generations of selection with imidacloprid, S. litura developed a 137.48-fold resistance to the insecticide. Bioassay revealed that this strain showed cross-resistance to acetamiprid (RR 8.52) and a little to lamdacyhalothrin (1.92) but negative cross-resistance was found to methomyl (−0.19). The resistant strain had a relative fitness of 0.38, with substantially lower rates of larval survival, larval duration, male pupal duration, development time, emergence rate of healthy adults, fecundity, hatchability, and prolonged larval and pupal duration. Mean relative growth rate of the larvae, intrinsic rate of population increase, and biotic potential was lower for the selected populations. The estimated realized heritability (h²) of imidacloprid resistance was 0.15 in the resistant strain of S. litura. Development of the resistance may cost significant fitness for the resistant population. This study provided valuable information for further understanding the impact of imidacloprid resistance on physiological parameters of S. litura and for facilitating the development of resistance management strategies.     

草地贪夜蛾对农药主要抗性机制的概述

草地贪夜蛾Spodoptera frugiperda是一种迁飞性害虫,源自美洲,已入侵非洲和亚洲部分国家并暴发成灾,2019年初入侵我国,严重威胁着我国玉米、小麦等粮食作物。目前草地贪夜蛾的防治主要依赖于化学农药和种植转基因作物。因长期使用农药,草地贪夜蛾已经对氨基甲酸酯类、有机磷酸酯类和拟除虫菊酯类等多种杀虫剂产生了不同程度的抗性。草地贪夜蛾对不同种类杀虫剂表现的抗性机制不同,主要有代谢抗性和靶标抗性2种。本文主要从代谢抗性和靶标抗性2个方面分别综述了草地贪夜蛾对不同种类杀虫剂的抗性机制,分析草地贪夜蛾对不同杀虫剂的抗性作用方式,并对草地贪夜蛾抗药性管理和监测进行展望。     

草地贪夜蛾为害玉米叶片程度的分级法及其在田间药效评价中的应用效果

为有效评价草地贪夜蛾 Spodoptera frugiperda的田间为害水平,采用5级法调查草地贪夜蛾在玉米植株上的为害程度,应用一元线性回归法分析5级法与其他分级法评估结果之间的关系,并评估该方法在田间药效评价试验中的应用效果。结果显示,采用叶片5级法与Williams 9级法评估的虫情指数之间、植株5级法与Davis 9级法评估的受害等级之间以及叶片5级法评估的虫情指数与植株5级法评估的受害等级之间均存在极显著线性关系,表明5级法可用于田间草地贪夜蛾为害水平的评价。植株受害程度较低时草地贪夜蛾低龄幼虫的出现频次较高,植株受害程度高时高龄幼虫的出现频次较高。不管是在未施药小区还是施药小区,采用叶片5级法调查的小区玉米倒一叶和倒二叶的平均受害等级及基于上部4片叶片的平均虫情指数均与小区总虫量之间存在显著线性关系,采用植株5级法调查的小区玉米植株整体平均受害等级与小区总虫量之间存在极显著线性关系。基于5级法调查的玉米倒一叶、倒二叶、植株整体平均受害等级推算的农药田间防治效果与基于虫量法推算的防治效果接近,说明5级法能用于评价农药的田间防治效果。     

草地贪夜蛾核型多角体病毒对草地贪夜蛾的毒力及其侵染特征

为明确草地贪夜蛾核型多角体病毒（Spodoptera frugiperda multiple nucleopolyhedrovirus,SfMNPV）对草地贪夜蛾Spodoptera frugiperda 的毒力及其体内外侵染特性,于室内测定SfMNPV对草地贪夜蛾的毒力,采用苏木精-伊红（hematoxylin-eosin,HE）染色分析SfMNPV侵染后草地贪夜蛾的组织病理学特征,通过实时荧光定量PCR（real-time fluorescence quantitative PCR,qPCR）技术分析SfMNPV在草地贪夜蛾体内和Sf9细胞系中的复制特征。结果表明,SfMNPV对草地贪夜蛾2龄幼虫的LC₅₀和LT₅₀分别为7.39×10⁵ OBs/mL和3.82 d;SfMNPV可侵染草地贪夜蛾幼虫的真皮、气管、脂肪体和中肠等组织,未侵染肌肉组织;SfMNPV侵染96 h后草地贪夜蛾体内SfMNPV基因组拷贝数最大,为3.91×10⁶ 拷贝数/ng,SfMNPV侵染96 h后Sf9细胞培养上清液中SfMNPV基因组拷贝数最大,为2.08×10⁷ 拷贝数/mL。     

陕西省兴平市玉米田草地贪夜蛾的发生动态及关键防治技术

为明确草地贪夜蛾Spodoptera frugiperda在陕西省关中地区的种群消长动态,筛选适合当地的抗虫玉米品种和高效防治药剂,于2020—2022年连续3年在兴平市玉米田采用性诱捕器监测其发生动态,采用植株受害5级法评估其对8个当地主栽玉米品种的为害及产卵选择,同时测定7种不同类型杀虫剂对当地种群卵和3龄幼虫的毒力及田间防效。结果表明,2020年和2021年草地贪夜蛾在兴平市的成虫始见期为6月中旬,2022年为7月下旬,连续3年的终见期均为11月上旬,高峰期均为8月下旬至9月下旬;其中在2021年的发生量最大,为212头/诱捕器。成虫产卵对玉米品种和叶片部位具有选择性,其中在郑单958上所产卵块数显著低于华农887,在叶片背面所产卵块数显著高于叶片正面。供试玉米品种中秦龙14和郑单958受害较轻,播后25~35 d平均受害级别均低于2.0;正大12和纪元1号受害最重,平均受害级别介于2.5~3.4之间。供试药剂中,5.7%甲维盐对草地贪夜蛾卵的毒力最高,LC₅₀为5.03 mg/L,其次为60 g/L乙基多杀菌素和10%虫螨腈,二者LC₅₀介于25.81~32.27 mg/L之间;5.7%甲维盐、60 g/L乙基多杀菌素和50 g/L虱螨脲对幼虫的毒力较强,三者LC₅₀介于0.14~0.63 mg/L之间。施用5.7%甲维盐、60 g/L乙基多杀菌素和50 g/L虱螨脲后3~10 d对草地贪夜蛾的田间防效最好,为92.34%~99.12%;其次为200 g/L氯虫苯甲酰胺和150 g/L茚虫威,药后3~10 d的田间防效为85.67%~92.09%。表明陕西省主栽玉米品种秦龙14和郑单958对草地贪夜蛾有一定抗性,5.7%甲维盐和60 g/L乙基多杀菌素对当地种群卵和幼虫的毒力最高且田间防效最好。     

草地贪夜蛾Bt抗性相关lncRNA的预测及分析

为明确长链非编码RNA（long non-coding RNA,lncRNA）在草地贪夜蛾Spodoptera frugiperda对Bt产生抗性中的作用,采用生物信息学预测草地贪夜蛾的lncRNA基因,并对其进行特征分析、差异表达分析、靶标基因预测以及功能富集分析。结果显示,草地贪夜蛾46个RNA-Seq数据中共有2 033个lncRNA基因,对应2 034条lncRNA转录本;与蛋白质编码基因相比,草地贪夜蛾lncRNA的外显子数量更少,转录本长度更短。草地贪夜蛾Bt抗性品系和敏感品系之间共有266个差异表达的lncRNA,其中128个上调表达,138个下调表达;在lncRNA靶标中,顺式调控和反式调控的靶标基因分别为2 129个和1 955个;靶标基因分子功能主要集中于有毒物质结合、水解酶和丝氨酸类蛋白活性等。10个差异表达的lncRNA靶向碱性磷酸酶、氨肽酶N、ABC转运蛋白、丝裂原活化蛋白激酶以及钙黏蛋白等抗性相关基因,表明lncRNA可能参与了草地贪夜蛾对Bt抗性的形成。     

Agrobacterium-mediated transformation efficiency is altered in a novel rice bacterial blight resistance cultivar and is influenced by environmental temperature

Y73 is a progeny of asymmetric somatic hybridization from an elite japonica rice cultivar (Dalixiang) and a wild rice (Oryza meyeriana), which shows high resistance to bacterial blight. It has a similar genetic background to its recurrent parent, Dalixiang, but Y73 has a high resistance to both bacterial blight and Agrobacterium. The transformation efficiency of Y73 was 35.7%, while Dalixiang had higher transformation efficiency (71.2%) under the same co-cultivation temperature (25 °C). These results indicate that the resistance to Agrobacterium tumefaciens in Y73 was linked with its characteristic of bacterial blight resistance, and was obtained from its donor parent, O. meyeriana. Further studies also showed that the resistance to A. tumefaciens was weakened at a lower temperature (20 °C). To study its molecular mechanism, the expression levels of genes associated with Agrobacterium-mediated transformation (OsMPKs, OsVIP1s and OsPR1s) in rice were investigated by Quantitative real-time PCR (qRT-PCR) analysis. The expression levels of OsMPK genes remained unchanged at different temperatures, while all OsVIP1s and almost all OsPR1s were up-regulated and transformation efficiency of Y73 was increased notably when infected by Agrobacterium at 20 °C compared with 25 °C. This study suggests that the bacterial blight resistance genes in Y73 also have an effect on Agrobacterium-mediated transformation and that the response to temperature is associated with the regulation of MAPK/VIP1 defense signaling pathway.     

Sulfoxaflor and the sulfoximine insecticides: Chemistry,mode of action and basis for efficacy on resistant insects

The sulfoximines, as exemplified by sulfoxaflor ([N-[methyloxido[1-[6-(trifluoromethyl)-3-pyridinyl]ethyl]-λ⁴-sulfanylidene] cyanamide] represent a new class of insecticides. Sulfoxaflor exhibits a high degree of efficacy against a wide range of sap-feeding insects, including those resistant to neonicotinoids and other insecticides. Sulfoxaflor is an agonist at insect nicotinic acetylcholine receptors (nAChRs) and functions in a manner distinct from other insecticides acting at nAChRs. The sulfoximines also exhibit structure activity relationships (SAR) that are different from other nAChR agonists such as the neonicotinoids. This review summarizes the sulfoximine SAR, mode of action and the biochemistry underlying the observed efficacy on resistant insect pests, with a particular focus on sulfoxaflor.     

Investigating nicotinic acetylcholine receptor expression in neonicotinoid resistant Myzus persicae FRC

The peach–potato aphid Myzus persicae is a pest of many commercial crops due to its polyphagous nature of feeding and has a well-documented history of acquiring resistance to insecticides. In 2009 a strain (M. persicae FRC) emerged in southern France with a point mutation (R81T) at the nicotinic acetylcholine receptor (nAChR), the target site for neonicotinoids such as imidacloprid. This point mutation was associated with the loss of the high affinity imidacloprid binding site (pM Kd), with the single remaining binding site (low nM Kd) highly overexpressed compared to laboratory controls (Bass et al., 2011 [1]). Here we report that after 2 years of continuous selection in the glass house environment with neonicotinoids, the total level of IMD-sensitive nAChRs (low nM Kd) in M. persicae FRC is now comparable to laboratory controls (pM and low nM Kd). Interestingly, despite this large reduction in IMD-sensitive nAChRs, this was not associated with any significant alteration in NNIC-lethality. Additionally, sustained absence of neonicotinoid-selection did not alter nAChR protein levels. We suggest that alterations in nAChR protein expression level described in the original characterisation of the field-isolated M. persicae FRC is unlikely to have been a direct consequence of the R81T mutation. Rather, we speculate that nAChR expression in aphids is likely influenced by as yet unknown conditions in the natural field environment that are absent in the laboratory setting.     

Molecular cloning and mRNA expression of a ryanodine receptor gene in the cotton bollworm, Helicoverpa armigera

Ryanodine receptors (RyRs) are the targets of novel diamide insecticides. The cotton bollworm, Helicoverpa armigera, is one of the most important cotton pests in the world. In this study, we report the full-length RyR cDNA sequence (named as HaRyR) of H. armigera. The 16,083-bp contiguous sequence encoded 5, 142 amino acid residues, which shares 80% and 78% overall identities with its homologues in Nilaparvata lugens (NlRyR) and Drosophila melanogaster (DmRyR), respectively. All hallmarks of RyR proteins are conserved in the HaRyR, including the GXRXGGGXGD motif conserved in the Ca²⁺ release channels and four copies of RyR domain unique to RyR channels. The previously identified seven lepidopteran-specific RyR residues were also found in HaRyR (N⁴⁹⁷⁷, N⁴⁹⁷⁹, N⁴⁹⁹⁰, L⁵⁰⁰⁵, L⁵⁰³⁶, N⁵⁰⁶⁸ and T⁵¹¹⁹). An amino acid sequence alignment showed that the N-terminal region of HaRyR (residues 188–295) shared high sequence identity with NlRyR (94%) and DmRyR (92%), and moderate sequence identity (47–50%) with three rabbit RyR isoforms, while the short segment of the C-terminal transmembrane region of HaRyR (residues 4632–4676) exhibited moderate sequence identity with NlRyR (69%) and DmRyR (67%), and low sequence identity (19–28%) with three rabbit RyR isoforms. In addition, expression analysis of HaRyR revealed that the mRNA expression level in eggs was significantly lower than in third instar larvae, pupae and adults, and anatomical regulation of HaRyR expression was also observed with the highest expression level in head compared with thorax and abdomen. Our results lay a foundation for comprehensive structural and functional characterization of HaRyR and for understanding of the molecular mechanisms of toxicity selectivity of diamide insecticides among different species.     

Toxicological and metabolic parameters of the teleost fish (Leporinus obtusidens) in response to commercial herbicides containing clomazone and propanil

Pesticides, such as herbicides can affect the metabolic and toxicological parameters on fish. For this reason, an experiment was carried out with the objective of to evaluate the effects of commercial formulations of clomazone and propanil herbicides on acetylcholinesterase (AChE), thiobarbituric acid-reactive substances (TBARS), catalase (CAT) and metabolic parameters in teleost fish (Leporinus obtusidens). Fish were exposed during 90 days to field measured concentration of the herbicides clomazone and propanil (376 and 1644 μg/L, respectively) on rice paddy water. Specific AChE activity in the brain and muscle decreased and TBARS levels decreased in brain, muscle and liver tissues. Liver catalase decreased after exposure to both herbicides. Metabolic parameters in the liver and white muscle showed different changes after exposure to both herbicides. In summary, the results showed that clomazone and propanil affects toxicological and metabolic parameters of piavas. These results suggest that environmentally relevant herbicides concentrations are toxic to Leporinus obtusidens.     

甜菜夜蛾对氯虫苯甲酰胺抗性种群选育及鱼尼丁受体基因表达特征

为明确甜菜夜蛾Spodoptera exigua对氯虫苯甲酰胺的抗性发展及抗性种群中鱼尼丁受体(ryanodine receptor,RyR)基因的表达量变化,室内采用饲料混毒法进行甜菜夜蛾对氯虫苯甲酰胺的抗性选育,荧光定量PCR技术研究了抗性和敏感种群之间RyR基因mRNA表达量的差异。结果表明,室内选育31代后获得一个抗性倍数为105.60倍的甜菜夜蛾抗性种群,其mRNA表达量在甜菜夜蛾不同发育阶段及抗性种群和敏感种群之间均存在差异,以表达量最低的卵期作为对照,抗性种群中1龄幼虫表达量最高,是卵期的154.58倍;其次是雄性成虫,其表达量是卵期的101.51倍;2~5龄幼虫分别是卵期的59.56、35.35、72.99和19.84倍。抗性种群中1、2和4龄幼虫mRNA表达量分别是敏感种群的5.99、2.79和2.14倍,其余阶段低于敏感种群。表明甜菜夜蛾对氯虫苯甲酰胺的抗药性可能主要表现在幼虫阶段,RyR基因的表达量变化与氯虫苯甲酰胺诱导有关。     

新烟碱类杀虫剂在韭菜中的内吸性及其对韭菜迟眼蕈蚊幼虫的毒力比较

通过药剂水培法,研究比较了不同培养条件对7种新烟碱类杀虫剂在韭菜中的内吸性及其对韭菜迟眼蕈蚊Bradysia odoriphaga(Diptera:Sciaridae)4龄幼虫的毒力。结果表明:随着水培时间的延长,韭菜中7种杀虫剂对供试幼虫的毒力均呈增强的趋势,其中在100 mg/L下,噻虫胺和吡虫啉产生毒力作用较快,水培72 h时对幼虫的致死率分别为32.38%和36.50%,168 h时,呋虫胺的致死率最高,达96.35%;同一药剂在相同浓度下处理,内吸至韭菜叶部产生的致死作用快于内吸至假茎所产生的作用,并且致死率更高;水培温度及药液浓度越高,内吸后毒力产生越快,且致死率越高。表明新烟碱类杀虫剂可以通过韭菜根部处理防治咀嚼式口器害虫韭菜迟眼蕈蚊。     

三种植物次生物质对B型烟粉虱羧酸酯酶活性及其对药剂敏感度的影响

采用人工饲料添加法,研究了不同浓度的槲皮素、2-十三烷酮和葫芦素B 3种植物次生物质对B型烟粉虱Bemisia tabaci羧酸酯酶(CarE)活性的影响,同时比较研究了用3种植物次生物质处理后敌敌畏、脱叶磷、灭多威、乐果和氧乐果5种药剂对CarE 的抑制中浓度(I50)的变化。结果表明: 0.01~1.0 mg/mL的槲皮素对B型烟粉虱成虫CarE活性均具有明显的诱导增加作用,最高为对照的4.32倍; 0.1和0.5 mg/mL的2-十三烷酮处理使CarE活性比对照分别下降了22%和58%; 0.75~30.0 mg/L的葫芦素B对烟粉虱CarE活性均表现为抑制作用;用1.0 mg/mL的槲皮素处理24 h后,敌敌畏和脱叶磷对烟粉虱CarE的I50值分别增加为对照的20.05和3.16倍; 3.75 mg/L 的葫芦素B处理后,敌敌畏和灭多威对CarE 的I50值分别增加为对照的25.94、2.45倍; 1.0 mg/mL的2-十三烷酮处理后敌敌畏、脱叶磷和乐果对CarE 的I50值分别降低了89%、83%和90%。     

The A302S mutation in Rdl that confers resistance to cyclodienes and limited cross-resistance to fipronil is undetectable in field populations of house flies from the USA

Fipronil is a relatively new insecticide with great potential for insect control, however widespread use of cyclodiene insecticides has selected for an A302S mutation in the Rdl (GABA gated chloride channel) allele. This mutation gives resistance to cyclodienes and limited cross-resistance to fipronil. Given the concern over the possible reduction in efficacy and/or lifetime that fipronil might be used for pest control (given the extensive use of cyclodienes in the past), it is important to know the frequency of the A302S Rdl mutation in field populations. To ascertain the relative frequency of the A302S Rdl mutation in house fly populations we used three experimental approaches. First, we attempted to select for fipronil resistance by initially treating 33,100 field collected flies and selecting 14 additional generations. We were unable to produce a highly resistant strain. Second, we directly sequenced field collected flies. Third, we tested field collected house flies with a diagnostic dose of dieldrin and then genotyped the survivors. Out of the 4750 flies tested, there were no Rdl resistance alleles detected. We conclude that the resistant Rdl allele is rare in house flies in the US due to decades without cyclodiene use and a fitness disadvantage (in the absence of cyclodienes) of the 302S Rdl allele. The limited cross-resistance provided by the cyclodiene resistant Rdl allele, combined with the very low frequency of this allele in field populations, suggests that fipronil could be a promising insecticide for house fly control.