氯虫苯甲酰胺对小菜蛾亚致死效应的研究

探讨亚致死剂量氯虫苯甲酰胺对小菜蛾2、3龄幼虫的亚致死效应,为氯虫苯甲酰胺的合理使用提供依据。采用药膜法确定氯虫苯甲酰胺对小菜蛾2龄和3龄幼虫的亚致死剂量(LC25、LC40),以试虫死亡率、取食量、化蛹时间、蛹重、羽化率为指标研究其对小菜蛾的亚致死效应。与对照相比,亚致死剂量氯虫苯甲酰胺处理的小菜蛾除死亡率显著增高外,幼虫生长明显受到抑制,幼虫至化蛹平均历期显著延长,取食量、蛹重和化蛹率明显降低。结果表明亚致死剂量氯虫苯甲酰胺对小菜蛾种群增长有一定抑制作用,对小菜蛾综合防治策略的制定有积极意义。     

亚致死浓度氯虫苯甲酰胺对小菜蛾生物学特性的影响

探讨亚致死剂量氯虫苯甲酰胺对小菜蛾3龄幼虫的亚致死效应,为氯虫苯甲酰胺的合理使用提供依据。采用叶片药膜法测定亚致死剂量氯虫苯甲酰胺对小菜蛾3龄幼虫生物学特性的影响。结果表明:与对照相比,亚致死浓度的氯虫苯甲酰胺处理后,小菜蛾4龄龄期和蛹期明显延长,化蛹率、羽化率、单雌产卵量明显降低。另外蛹重也明显低于对照。这表明亚致死剂量氯虫苯甲酰胺对小菜蛾种群增长有一定抑制作用,对小菜蛾综合防治策略的制定有积极意义。     

小菜蛾对丁烯氟虫腈的抗性选育及生物适合度

用丁烯氟虫腈对小菜蛾敏感品系在室内经7代次药剂汰选,获得了抗性品系(R),与敏感品系(S)比较其抗性指数为77.58倍。抗性汰选前后,采用点滴法测定的结果表明,小菜蛾对阿维菌素、毒死蜱、灭多威、杀虫单和高效氯氰菊酯的敏感性并无明显变化,其抗性指数为0.80~1.36,均不存在交互抗性。通过构建种群生命表,观察比较了小菜蛾抗丁烯氟虫腈品系和敏感品系的一系列生长发育和繁殖特征。结果表明,抗丁烯氟虫腈品系较敏感品系幼虫期和蛹历期延长,低龄幼虫龄期转换时的虫口死亡率增加,蛹重减轻,化蛹率和羽化率降低;在繁殖上表现为产卵量增加,而孵化率和雌雄比降低,雌成虫的数量、寿命减少。抗丁烯氟虫腈种群相对于敏感种群的相对适合度为0.84,抗性种群在繁殖能力上存在明显的生存劣势。     

小菜蛾CYP6BG1基因体外真核表达及其对氯虫苯甲酰胺的代谢

小菜蛾Plutella xylostella是为害十字花科作物的世界性重要害虫,对氯虫苯甲酰胺产生了严重抗性。已经明确小菜蛾CYP6BG1的过量表达与其对氯虫苯甲酰胺的抗性相关。本文进一步通过昆虫杆状病毒表达系统在Sf9细胞中表达了小菜蛾CYP6BG1和NADPH-细胞色素P450还原酶(CPR)蛋白,并检测Sf9细胞表达CYP6BG1蛋白后对氯虫苯甲酰胺的敏感变化及氯虫苯甲酰胺经CYP6BG1蛋白代谢后对小菜蛾幼虫毒力的变化。结果显示:Sf9细胞中与CPR共表达的CYP6BG1具有7-乙氧基香豆素O-脱乙基酶活性,且细胞对氯虫苯甲酰胺的敏感性显著下降。将表达的蛋白与不同浓度的氯虫苯甲酰胺孵育24 h后,对3龄小菜蛾幼虫致死率显著低于对照组。本研究结果间接表明CYP6BG1能够降解氯虫苯甲酰胺,从而增强小菜蛾对氯虫苯甲酰胺的抗性。     

氯虫苯甲酰胺对小菜蛾鱼尼丁受体基因mRNA表达量的影响

采用实时荧光定量PCR研究比较了氯虫苯甲酰胺、甲氨基阿维菌素苯甲酸盐和溴虫腈对小菜蛾鱼尼丁受体基因mRNA表达量的影响,研究分析了低剂量氯虫苯甲酰胺连续处理和间断处理对小菜蛾鱼尼丁受体基因表达量的影响,以及对氯虫苯甲酰胺产生抗性的小菜蛾与敏感小菜蛾鱼尼丁受体基因表达量的差异。结果表明:氯虫苯甲酰胺可提高小菜蛾鱼尼丁受体基因表达量,而甲氨基阿维菌素苯甲酸盐和溴虫腈对其表达量均无影响;0.05和0.01 mg/L的氯虫苯甲酰胺均可使小菜蛾鱼尼丁受体基因表达量增加;对氯虫苯甲酰胺产生抗性的小菜蛾鱼尼丁受体基因表达量是敏感小菜蛾的5.79倍。研究结果有助于阐明氯虫苯甲酰胺的作用机理,并可能为筛选作用于鱼尼丁受体的新化合物提供参考。     

四种P450基因调控小菜蛾对氯虫苯甲酰胺的抗性

为研究CYP6家族P450基因在小菜蛾Plutella xylostella代谢氯虫苯甲酰胺中的作用,利用浸叶法测定不同小菜蛾种群3龄幼虫对氯虫苯甲酰的抗性水平,采用实时荧光定量PCR(quantitative realtime PCR,q RT-PCR)方法分析CYP1v3、CYP1v4、CYP6B6和CYP6f这4种P450基因在小菜蛾不同抗性种群体内的表达差异及杀虫剂的短期诱导效应,并通过RNA干扰技术沉默4种P450基因后分析小菜蛾对氯虫苯甲酰胺的敏感性。结果显示,4种P450基因在中等抗性水平小菜蛾种群体内高表达;氯虫苯甲酰胺处理可诱导4种基因显著上调表达。分别沉默4种P450基因后,处理组小菜蛾的P450酶活力显著下降37.60%～54.82%,且处理组小菜蛾的死亡率显著高于对照组;同时沉默4种基因处理组的小菜蛾P450酶活力显著低于其他各处理组。表明4种P450基因可能同时参于小菜蛾对氯虫苯甲酰胺的代谢。     

张家口地区田间小菜蛾种群对6种新型杀虫剂的敏感性测定

用浸液法测定了河北省张家口地区田间小菜蛾种群对氯虫苯甲酰胺等几种新型杀虫剂的敏感性。田间小菜蛾种群对甲氨基阿维菌素苯甲酸盐、多杀菌素、茚虫威、氯虫苯甲酰胺、三氟甲吡醚、氰氟虫腙等的抗性倍数分别为5.19、2.22、0.25、1.07、1.16和0.92倍。敏感性测定结果表明,田间种群对甲氨基阿维菌素苯甲酸盐产生了低水平抗性,对多杀菌素、氯虫苯甲酰胺、三氟甲吡醚未产生明显抗药性,对茚虫威、氰氟虫腙相当敏感。     

我国部分地区田间小菜蛾种群对8种常用杀虫剂的抗性检测

采用浸叶法测定了来自5个地区的田间小菜蛾种群对8种杀虫剂的抗性水平。结果表明,5个地区小菜蛾种群对氯虫苯甲酰胺均产生了中等水平以上的抗性,江苏无锡和广东增城种群的抗性达1428.16倍和6642.12倍,为极高抗性水平;对丁醚脲均处于敏感或敏感性降低水平。5个地区的小菜蛾种群对甲氨基阿维菌素、虫螨腈、S-茚虫威、虫酰肼和唑虫酰胺等5种药剂的敏感性各异,大多处于敏感或敏感性降低水平,但也有些地区产生了中等水平的抗性。本研究结果明确了我国部分地区对小菜蛾抗药性情况,可为小菜蛾的抗性治理提供参考。     

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。     

广东草地贪夜蛾不同田间种群对4种杀虫剂的抗药性监测

采用饲料浸药法测定了草地贪夜蛾对氯虫苯甲酰胺、多杀霉素、甲氨基阿维菌素苯甲酸盐、虫螨腈的相对敏感基线,并测定了广东省广州、东莞、开平、平远和阳江等5地田间草地贪夜蛾种群对氯虫苯甲酰胺、多杀霉素、甲氨基阿维菌素苯甲酸盐和虫螨腈的抗药性。结果表明：上述4种药剂对敏感草地贪夜蛾种群2龄幼虫的LC₅₀在0.037～0.300 6 mg/L之间。除平远种群对氯虫苯甲酰胺保持敏感外,其余4个种群对氯虫苯甲酰胺抗性在3.51～9.99倍之间,属低水平抗性;阳江和东莞种群对多杀霉素的抗性分别为17.62倍和17.71倍,属中等水平抗性;其他3个种群对多杀霉素抗性在4.69～7.60倍之间,属低水平抗性;除平远草地贪夜蛾种群对甲氨基阿维菌素苯甲酸盐抗性倍数为5.34倍,属低水平抗性外,其他4个草地贪夜蛾种群对甲氨基阿维菌素苯甲酸盐抗性倍数在0.39～1.08倍之间,属于敏感水平;开平草地贪夜蛾种群对虫螨腈的抗性倍数为10.77倍,属中等水平抗性;广州、东莞和阳江草地贪夜蛾种群对虫螨腈的抗性倍数在4.21～8.84倍之间,属低水平抗性;平远草地贪夜蛾种群对虫螨腈的抗性倍数为2.37倍...     

小菜蛾对唑虫酰胺的抗性监测、抗性生化机制及交互抗性

为了明确小菜蛾Plutella xylostella对唑虫酰胺的抗性特征,采用生物生化方法测定了江西省5个蔬菜产区小菜蛾田间种群对唑虫酰胺的抗性水平,并研究了小菜蛾唑虫酰胺抗性品系对其它药剂的交互抗性和生化抗性机制。结果显示,分宜县和高安市小菜蛾田间种群对唑虫酰胺尚未产生明显抗性,永丰县、德安县和余江县小菜蛾种群对唑虫酰胺产生了低水平的抗性,抗性倍数为5.20~8.20倍;小菜蛾唑虫酰胺抗性品系对阿维菌素、氯虫苯甲酰胺、氟虫双酰胺和茚虫威有中低水平的交互抗性,抗性倍数分别为11.72、3.44、2.77和2.20倍,而对溴虫腈、定虫隆和丁醚脲无交互抗性;增效剂磷酸三苯酯和胡椒基丁醚对小菜蛾唑虫酰胺抗性品系均有显著增效作用,增效倍数分别为3.42倍和2.64倍;唑虫酰胺抗性品系的酯酶和多功能氧化酶活性均显著提高,分别为敏感品系的2.18倍和1.64倍。研究表明,小菜蛾对唑虫酰胺产生抗性可能与酯酶和多功能氧化酶活性的升高有关。     

小菜蛾中肠氨肽酶N2在昆虫细胞中的表达

利用昆虫细胞-杆状病毒表达系统(Bac-to-Bac baculovirus expression system)表达获得小菜蛾 Plutella xylostella(L.)中肠氨肽酶N2(aminopeptidase N,APN2)蛋白,成功建立了该蛋白在昆虫细胞中的表达体系。将对Cry1Ac毒素敏感和已产生抗性的小菜蛾中肠APN2基因插入表达载体pFAST Bac HTB中,并在草地夜蛾 Spodoptera frugiperda 细胞系(Sf9)中进行了重组表达。聚丙烯酰胺凝胶电泳(SDS-PAGE)及蛋白免疫印迹技术(Western-blotting)结果显示,在107 kDa处有1条特异性蛋白条带。研究表明,小菜蛾中肠APN2基因可在Sf9细胞中成功表达,这为继续研究其功能及抗性的关系奠定了基础。     

Reduced fitness associated with spinosad resistance in <Emphasis Type="Italic">Helicoverpa armigera</Emphasis>

The fitness cost of spinosad resistance was investigated in the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae). Laboratory experiments were conducted to compare relative fitness of H. armigera between the spinosad-susceptible and -resistant strains. During the experiments, the average development periods of the resistant strain were lengthened by 4–5 days, reflected in a prolongation of egg, larval and pupal periods. Furthermore, pupal survival, pupal weight, the mean life span of emerged adults, eggs laid and hatched decreased greatly in the resistant strain in comparison with the susceptible strain. Other life-cycle parameters such as larval survival, larval wet weights, prepupal periods, pupation ratio, and sex ratio did not change significantly. As a result, both net replacement rate (R ₀) and intrinsic rate of increase (r _m) were reduced for the resistant strain. Our results clearly indicated that relative fitness of resistant individuals was reduced in the absence of spinosad. Rational measures including pesticide rotations should be expected to delay development of resistance to spinosad in H. armigera field populations from China.     

The stability and biochemical basis of fufenozide resistance in a laboratory-selected strain of Plutella xylostella

Fufenozide, a novel non-steroidal ecdysone agonist, exhibits good efficacy and plays an important role in controlling lepidopteran pests in China. A high level of resistance to fufenozide (320.3-fold) in Plutella xylostella (L.) has been selected under laboratory conditions. In this study, the stability and biochemical basis of resistance to fufenozide were investigated. The resistance ratio to fufezonide declined substantially from 320.3- to 20.5-fold in the fufenozide resistant strain (JSR) when the selection pressure was removed for six consecutive generations. However, the susceptibility to fufezonide could not be fully restored after 28 generations without exposure to the compound. Increased relative fitness (0.87) was associated with the rapid reversal of resistance in the resistance decaying strain (JSRD). The enzymatic analyses showed reduced specific activities of monooxygenase (MO), glutathione S-transferases (GSTs), carboxylesterase and aryl-acylamidase in the JSRD strain, whereas the activity of chitinase increased. The current findings suggest the selected resistance to fufenozide in P. xylostella is not stable, and the four detoxifying enzymes including MO, GSTs, carboxylesterase, and aryl-acylamidase may be involved. For resistance management proposes, insecticide rotation should be a viable option to delay the occurrence of fufenozide resistance in the field.     

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.     

Residual toxicity and sublethal effects of chlorantraniliprole on Plutella xylostella (lepidoptera: plutellidae)

BACKGROUND: The diamondback moth (DBM), Plutella xylostella (L.), is the most important pest of cruciferous vegetables in the world. Chlorantraniliprole is a novel anthranilic diamide insecticide registered for the control of lepidopteran pests. The dose response, residual toxicity and sublethal effects of chlorantraniliprole applied for 48 h at LC₁₀ (0.02 mg L^?1) and LC₂₅ (0.06 mg L^?1) on P. xylostella were investigated. RESULTS: Leaf‐dip bioassays showed that chlorantraniliprole had a high level of toxicity against larvae of P. xylostella, and the 48 h LC₅₀ values were 0.23 and 0.25 mg L^?1 for a susceptible and field strain respectively. Chlorantraniliprole also had a long‐lasting effect when the larvae were exposed to chlorantraniliprole field sprayed on radish seedlings. Sublethal effects of chlorantraniliprole were indicated by reduced pupation, pupal weight and adult emergence rates. There was also an increase in the duration of female preoviposition period, decreased fecundity and egg hatch and decreased survival rates of the offspring. The mean values of the net reproductive rate (R₀), intrinsic rate of increase (r_m), finite rate of increase (λ) were significantly lower in the treatment than in control groups. CONCLUSIONS: These results indicate that chlorantraniliprole is effective against P. xylostella. The sublethal concentrations of chlorantraniliprole may reduce the population growth of P. xylostella by decreasing its survival and reproduction, and by delaying its development. Copyright © 2012 Society of Chemical Industry     

Cross-resistance patterns and fitness in fufenozide-resistant diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae)

BACKGROUND: Fufenozide is a novel non‐steroidal ecdysone agonist with good efficacy against diamondback moth (DBM), Plutella xylostella (Lepidoptera: Plutellidae). At present, it is widely applied for the control of a range of lepidopterous pests in China. This study compared the activities of fufenozide and 12 other insecticides against unselected and fufenozide‐selected strains of DBM to examine potential patterns of cross‐resistance. The relative fitness of the fufenozide‐selected strain was assessed to provide information pertinent to insecticide resistance management. RESULTS: Compared with the susceptible strain (JSS), the fufenozide‐resistant strain (JSR) showed high cross‐resistance to dibenzoylhydrazines and benzoylphenylureas, low cross‐resistance to abamectin and no cross‐resistance to organophosphates, carbamates and pyrethroids. JSR had a lower reproductive ability and a relative fitness of 0.5 compared with JSS. CONCLUSION: P. xylostella has the potential to develop resistance to fufenozide, albeit at the expense of fitness. Cross‐resistance between the same and other classes of insecticides is of concern, and should be a key consideration when implementing fufenozide‐based control strategies for this species. Copyright © 2011 Society of Chemical Industry