Management of diamondback moth,Plutella xylostella (Lepidoptera: Plutellidae) by mating disruption

Abstract Field trials were conducted in China in 2008 and 2009 to evaluate the efficacy of mating disruption (MD) on diamondback moth, Plutella xylostella, in cabbage, Brassica oleracea var. capitata. Effectiveness was positively correlated with the MD dispenser density in the field. A density of 167 MD dispensers per ha produced an average population decrease of about 50% compared to the conventional‐practice field. Significant fewer males were captured in pheromone‐treated and conventional‐practice fields than in the blank control field, but the difference was not significant between the pheromone‐treated and conventional‐practice fields. In addition, fewer eggs and larvae were observed in pheromone‐treated fields. Our results suggest mating disruption coupled with minimal insecticidal supplements is a promising solution for resistance management and control of diamondback moth infestation.     

羧酸酯酶介导的小菜蛾对氟虫腈的抗性

【目的】羧酸酯酶（carboxylesterases, CarEs）是昆虫重要的解毒代谢酶之一,可以介导靶标昆虫对多种杀虫剂的代谢抗性。本研究检测了羧酸酯酶对小菜蛾Plutella xylostella 抗药性的介导功能,旨在阐明羧酸酯酶在小菜蛾代谢解毒中的生理生化和分子机理。【方法】采用点滴法测定氟虫腈对小菜蛾敏感种群和抗氟虫腈种群的毒力,以及羧酸酯酶抑制剂磷酸三苯酯（triphenyl phosphate, TPP）对氟虫腈的增效作用;以LC₃₀和LC₅₀浓度的氟虫腈处理抗性小菜蛾,测定药剂处理后CarEs酶活性的变化;利用qRT-PCR技术分析Pxae22和Pxae31两个基因在小菜蛾不同发育阶段、组织和种群的表达模式;利用dsRNA干扰Pxae22和Pxae31后观察基因的表达变化和小菜蛾3龄幼虫对药剂敏感性的变化。【结果】TPP可以削弱小菜蛾3龄幼虫对氟虫腈的抗性,增效倍数约为6倍;使用较低剂量（LC₃₀和LC₅₀）氟虫腈处理小菜蛾3龄幼虫后,处理组CarEs比活力明显高于对照,提示氟虫腈对小菜蛾CarEs活性具有诱导作用。对羧酸酯酶基因Pxae22和Pxae31在小菜蛾不同发育阶段、4龄幼虫不同组织和不同种群3龄幼虫中的表达模式分析发现,这两个基因在小菜蛾4龄幼虫中的表达量最高;在4龄幼虫中以中肠组织中的表达量较高,头、表皮、脂肪体中的表达量很低; 抗性种群中的表达量显著高于敏感种群。通过干扰 Pxae22和 Pxae31后的qRT-PCR验证,两个基因的表达量均显著降低,进一步的氟虫腈毒力测定发现,干扰P xae22和 Pxae31后的小菜蛾3龄幼虫对氟虫腈的敏感性分别增加了1.63倍和1.73倍。【结论】羧酸酯酶在小菜蛾对氟虫腈解毒代谢中具有重要作用;Pxae22和Pxae31是小菜蛾的两个抗性相关基因,其表达水平的变化直接影响小菜蛾对氟虫腈的敏感性。     

Oviposition site selection by the diamondback moth,Plutella xylostella (L.) (Lepidoptera: Plutellidae)

AlthoughPlutella xylostella (L.) is a worldwide pest of cruciferous crops, relatively little is known about its oviposition behaviour. This study was undertaken to provide necessary information about mechanisms involved inP. xylostella host selection. Four oviposition behaviours were described. Moths were given artificial substrates treated with water, sinigrin, orBrassica napus (cv. Westar) squashes, combined withB. napus volatiles and/or grooves in the substrate. No eggs were deposited in the absence of olfactory and gustatory stimuli. Moths given gustatory but not olfactory stimuli deposited similar numbers of eggs but spent significantly more time performing olfactory-related behaviours. Conversely, moths given olfactory but not gustatory stimuli did not oviposit.SSubstrate grooves did not influence egg numbers but appeared to influence egg location. The order in which oviposition behaviours occurs and the relative importance of stimuli type are discussed.     

Functional and genetic characteristics of Chlorantraniliprole resistance in the diamondback moth,Plutella xylostella (Lepidoptera: Plutellidae)

The diamondback moth (Plutella xylostella) is a globally distributed and important economic pest, and it has developed resistance to all conventional insecticide classes used in the field. Chlorantraniliprole is a new chemical class of insecticide that acts as a conformation‐sensitive activator of the insect ryanodine receptor (RyR). In the present study, a field strain (16.3‐fold resistance to chlorantraniliprole) was collected in Korea and lab‐selected with chlorantraniliprole for more than one year. The resulting strain presented 2,157‐fold resistance to chlorantraniliprole. A point mutation (G4946E) in the RyR gene was observed at a high frequency in the resistant strain. Enzyme assays indicated that glutathione S‐transferase (GST) and P450 activity in the resistant strain were 2.4‐ and 1.96‐times higher than that of the susceptible strain, respectively. The expression of the RyR, GST (sigma, omega, and zeta) and CYP321E1 gene was higher in the resistant strain than in the susceptible strain. The F₁ progeny resulting from reciprocal crosses did not reveal maternal effects or a diamide‐susceptible phenotype, which suggests an autosomal nearly recessive mode of inheritance. In addition, we surveyed the susceptibility to 13 insecticides (3 diamides, 2 synthetic pyrethroids, 2 spinosyns, 1 organophosphate, 1 oxadiazine, 1 avermectin, and 3 others) in the chlorantraniliprole‐resistant strain. The resistant strain exhibited high cross‐resistance to flubendiamide (5,910 fold) and showed no cross‐resistance to spinetoram, spinosad, indoxacarb, and metaflumizone. These results can serve as an important basis for guiding the use of insecticides in the field.     

小菜蛾鞣化激素基因克隆及其功能分析

鞣化激素是调节昆虫表皮骨化和翅膀发育的一种神经激素, 尽管已经在许多不同种昆虫上克隆了鞣化激素基因, 但是关于小菜蛾 Plutella xylostella鞣化激素及其基因的研究至今未见报道。本研究克隆了两个小菜蛾鞣化激素基因Pxbursα和Pxbursβ （GenBank 登录号分别为KF498645和KF498646）全长cDNA, 其序列长度分别为537 bp和360 bp, 与已报道的其他昆虫的鞣化激素氨基酸序列一致性分别为51%~68% 和37%~57%。实时定量PCR分析发现Pxbursα和Pxbursβ均在蛹期表达量高, 而在幼虫期和成虫期的表达量低。以Pxbursα部分序列的双链RNA（dsRNA）饲喂小菜蛾4龄末期幼虫, 发现蛹期Pxbursα的表达受到了显著抑制, 小菜蛾的发育停滞在蛹期而无法正常羽化, 并最终死亡。由此推测, 小菜蛾鞣化激素基因在蛹期的大量表达对其生长发育和羽化具有重要的作用。     

光周期对小菜蛾羽化节律的影响

【目的】本研究旨在探究光周期对小菜蛾Plutella xylostella(Linn.)羽化节律的调控作用,为小菜蛾的季节性种群动态预测提供理论基础。【方法】在自然光周期和逆转光周期条件下,观察小菜蛾每日的羽化情况。【结果】在自然光周期和逆转光周期条件下,小菜蛾蛹在24 h均可以羽化。正常光周期下,羽化行为表现为近似抛物线形昼夜节律,雌成虫通常在黑暗阶段羽化（18:00-6:00）,而雄成虫大多在光照阶段羽化（6:00-18:00）。在逆转光周期下,小菜蛾蛹的羽化无明显规律。在正常光周期下,雌成虫的羽化频率在光期和暗期没有显著差异,但是,相比于暗期,雄成虫在光期羽化率较高;在逆转光周期后,雌成虫在暗期（6:00-18:00）的羽化数量比光期（18:00-6:00）高,雄成虫暗期的羽化数量比光期低。【结论】光周期逆转后小菜蛾雌成虫和雄成虫的羽化节律明显改变,这将有助于阐明害虫光周期变化与生物钟之间的关系。     

利用RNAi技术沉默小菜蛾类钙粘蛋白基因

RNA干扰（RNA interference, RNAi）是一种调控基因表达的方法, 其通过体外合成一段与内源靶基因同源的双链RNA（dsRNA）或siRNA, 导入生物体内, 使内源靶基因中同源mRNA降解, 从而达到阻抑基因表达的目的。类钙粘蛋白（cadherin-like protein）是位于昆虫中肠刷状缘膜囊(brush border membrane vesicles, BBMV)上与钙粘蛋白(cadherin)结构相似的物质, 是多种昆虫体内Bt杀虫蛋白的受体。本研究利用基因特异引物通过RT-PCR扩增了小菜蛾类钙粘蛋白基因的2个片段（CAD1和CAD2）, 合成相对应的双链RNA（double-stranded RNA, dsRNA）; 并将dsRNA通过显微注射导入小菜蛾3龄幼虫体内, 测定了不同靶位点、不同剂量、不同检测时间对目的基因mRNA表达量的影响。结果表明: 将70 nL CAD1对应的dsRNA注射到幼虫体内48 h后, 基因表达量显著下降, 72 h后恢复。免疫印迹检测结果表明, 类钙粘蛋白在注射dsRNA 48 h后幼虫BBMV中的含量明显下降。本实验成功实现了小菜蛾类钙粘蛋白基因的沉默, 该体系的成功建立为利用RNAi技术分析小菜蛾及其他鳞翅目昆虫基因的功能提供了参考。     

小菜蛾幼虫肠道细菌桃色欧文氏菌的代谢表型组学分析

[目的]肠道细菌桃色欧文氏菌Erwinia persicina是小菜蛾Plutella xylostella幼虫肠道的优势细菌,本研究旨在阐明桃色欧文氏菌的代谢表型特征.[方法]采用BIOLOG细胞表型芯片技术系统研究了桃色欧文氏菌的细胞表型;采用PM l-10代谢板,对桃色欧文氏菌的950种代谢表型进行了测定.[结果]桃色欧文氏菌能代谢39.47％的碳源、89.74％的氮源、100％的硫源和100％的磷源;高效代谢的碳源为有机酸类和碳水化合物类,高效代谢的氮源为氨基酸类.该肠道细菌表现出261种不同的氮源代谢通路和95种生物合成通路.桃色欧文氏菌具有广泛的适应性,能在分别具有高达9％氯化钠、4％氯化钾、5％硫酸钠、20％乙二醇、6％甲酸钠、2％尿素、6％乳酸钠、200mmol/L磷酸钠(pH 7.0)、20 mmol/L苯甲酸钠(pH 5.2)、100 mmol/L硫酸铵(pH 8.0)、100 mmol/L硝酸钠和100 mmol/L亚硝酸钠的渗透溶液中正常代谢;其适应pH值范围为4.5 ～10,最适7.0.在多种氨基酸的作用下,桃色欧文氏菌均表现出脱羧酶和脱氨酶活性.[结论]桃色欧文氏菌的代谢特征增加了我们对该肠道细菌,特别是其与宿主昆虫的互作及其在肠道环境中的适应性的认识,同时表明通过降低桃色欧文氏菌密度来防治小菜蛾的可能性.     

氰氟虫腙对小菜蛾阿维菌素抗性和敏感种群的亚致死效应(英文)

在实验室条件下采用生命表技术研究了氰氟虫腙亚致死剂量（LC₂₅）对小菜蛾Plutella xylostella阿维菌素抗性（AV-R）和敏感（AV-S）种群的亚致死效应, 旨在为小菜蛾对阿维菌素的抗性治理提供理论基础。结果表明： 氰氟虫腙对小菜蛾3龄幼虫抗性种群的LC₅₀和LC₂₅分别为0.24 mg/L和0.09 mg/L; 对敏感种群的LC₅₀和LC₂₅分别为0.20 mg/L和0.07 mg/L。氰氟虫腙亚致死剂量0.09 mg/L 处理小菜蛾后, 对处理代的影响表现为显著降低处理种群的化蛹率、 蛹重、 羽化率、 繁殖力; 明显延长蛹期, 缩短成虫产卵期和寿命; 对子代种群的影响表现为显著降低卵的孵化率、 幼虫各龄期的存活率, 延长发育历期。处理种群的内禀增长率（r_m）、 周限增长率（λ）和净增值率（R₀）显著低于对照种群（P<0.0001）。亚致死剂量的氰氟虫腙对小菜蛾抗性种群的影响大于敏感种群, 对处理代种群的影响大于子代种群。氰氟虫腙亚致死剂量可以极大地影响小菜蛾尤其是阿维菌素抗性种群的种群动态, 因此氰氟虫腙对于小菜蛾的抗性治理具有积极的作用。     

Reproductive morphology,copulation, and inter-populational variation in the diamondback moth,Plutella xylostella (L.) (Lepidoptera: Plutellidae)

Copulatory mechanisms and internal reproductive systems of male and female Plutella xylostella (L.) were investigated. Both male and female specimens exhibited characteristics typical of ditrysian Lepidoptera, with some peculiarities. Female structures appear to be relatively simple: the bursa copulatrix lacks cuticular signa within, the spermatheca lacks lagenar arms exhibited in some Lepidoptera, and colleterial glands have secretory system and reservoir combined. Male accessory gland ducts are joined distally, a condition that has not been described in other Lepidoptera. Genitalia of both sexes appear simple; females possess a posterior cuticular extension of abdominal sternites that houses the bursal duct and accomodates the needle-like aedeagus of a male during copulation. Despite the apparent simplicity, configuration of genitalia is highly specific in shape, angle, and size, which is likely to be important in maintaining reproductive isolation within the species. However, a population of P. xylostella from Australia showed some modification to both aedeagal and bursal structures. These variations suggest some important implications for considering species with worldwide distributions.     

Identification of the Diadegma species (Hymenoptera: Ichneumonidae, Campopleginae) attacking the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae)

The species of Diadegma that attack Plutella xylostella (Linnaeus) are revised. Following a morphometric study involving principal components and discriminant analyses, seven distinct morphospecies are recognized. One species is described as new: D. novaezealandiae from New Zealand. Diadegma mollipla (Holmgren) is the name for the species from sub-Saharan Africa and some Indian Ocean and South Atlantic islands. Diadegma varuna Gupta syn. nov. and D. niponica Kusigemati syn. nov. are both synonymized with D. fenestrale (Holmgren). Diadegma xylostellae Kusigemati is strongly presumed to be a synonym of D. semiclausum (Hellén). An illustrated identification key is provided and each species is described in a standard way.     

Occurrence of diamondback moth,Plutella xylostella (Lepidoptera: Plutellidae), and its larval parasitoids across Saskatchewan,Canada

The prevalence of the diamondback moth and its larval parasitoids, was explored across Saskatchewan, Canada. Higher numbers of diamondback moths were recorded early in the season than later and in the Mixed Grassland compared to three other ecoregions. The highest proportion of parasitism was observed in the Boreal Transition ecoregion in early season.     

Using yellow rocket as a trap crop for diamondback moth (Lepidoptera: Plutellidae)

Yellow rocket, Barbarea vulgaris (R. Br.) variety arcuata, was evaluated as a trap crop for diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), in cabbage, Brassica oleracea L. variety capitata, in 2003 and 2004. In 2003, the numbers of P. xylostella larvae found in field plots of cabbage alone were 5.2-11.3 times higher than those on cabbage plants in plots that included cabbage and several rows of yellow rocket. In an outdoor experiment in screenhouses, P. xylostella oviposition on cabbage was compared among six treatments that varied in the percentage of yellow rocket in relation to cabbage (0, 4, 8, 16, 24, and 32% of the plants were yellow rocket). Results indicated that the percentage of eggs laid on cabbage decreased as the percentage of yellow rocket in the treatment increased, but this decrease was not significant beyond 20% of the plants being yellow rocket. In 2004, the numbers of P. xylostella larvae in field plots of cabbage alone were 1.6-2.4 and 1.7-2.8 times higher than numbers in treatments with 10 and 20% trap crop, respectively. Sticky trap and sweep net captures of P. xylostella adults indicated that within-field dispersal was reduced by the presence of yellow rocket and aggregation occurred around yellow rocket plants. Our study suggests that using yellow rocket as a trap crop may reduce P. xylostella infestations in cabbage fields, and this possibility is discussed in the context of general crop and insect pest management practices in crucifers.     

利用寄主干扰效应防控小菜蛾田间种群的有效性

【目的】多寄主对植食性昆虫寄主选择行为具干扰效应。为探究在田间相似寄主混作是否能引起小菜蛾Plutella xylostella (L.)在寄主上的选择困难,减少在寄主上的产卵量,从而实现保护寄主植物的目标。【方法】本研究利用小菜蛾喜好性高的两种寄主（菜心Brassica campestris L.和芥菜B. juncea Coss）为试验对象,通过寄主植物单作、混作及不同配比与间隔种植等田间试验设计,研究了多寄主对小菜蛾寄主选择行为的干扰效应。【结果】在2年的田间试验中,两寄主混作处理上的虫量均低于单一寄主种植;2012年田间种群生命表数据显示,两种寄主田间混作处理上的小菜蛾种群趋势指数（I =4.797）低于单一寄主种植,菜心与芥菜单一种植处理上的I值分别为6.549与6.858;当菜心与芥菜这两种小菜蛾喜好性相近寄主同时存在时,对小菜蛾的干扰程度与两寄主的搭配比例有关,随着菜心所占比重的增大,田间虫量呈现先减后增的抛物线变化趋势;但两寄主间距在2 m内时,不同种植间隔对小菜蛾虫量变化无差异(F_3,8=0.204,P>0.05)。【结论】结果表明,在田间多寄主种植中,除了可利用陷阱作物来防控小菜蛾外,在相邻两畦地间以适当比例同时种植两种小菜蛾喜好性相近的寄主,利用两种寄主植物对植食性昆虫的寄主选择行为产生的干扰效应,也能有效影响田间小菜蛾的种群动态。     

Seasonal abundance of the parasitoid complex associated with the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae) in Hangzhou, China

An investigation of insect parasitoids of the diamondback moth, Plutella xylostella (Linnaeus), in brassica vegetable crops in the suburbs of Hangzhou was conducted during five periods from 1989 to 1997. Eight species of primary parasitoids were recorded: Trichogramma chilonis Ishii, Cotesia plutellae Kurdjumov, Microplitis sp., Oomyzus sokolowskii Kurdjumov, Diadromus collaris (Gravenhorst), Itoplectis naranyae (Ashmead), Exochus sp. and Brachymeria excarinata Gahan. Seven species of hyperparasitoids were also collected. Rates of parasitism of eggs of P. xylostella were usually very low. However, rates of parasitism of larvae and pupae were substantial and showed two peaks each year, around June-July and September-November respectively. Rates of parasitism during peaks were usually 10-60% and reached over 80% on a few occasions. Cotesia plutellae, O. sokolowskii and D. collaris were the major larval, larval-pupal and pupal parasitoids respectively. In the field, C. plutellae was active throughout the year. Oomyzus sokolowskii was active from May to October, entered a quiescent pupal stage in October-November to overwinter and did not emerge until next April-May. Diadromus collaris was recorded from April to July and October. Rates of parasitism of P. xylostella in radish and mustard fields were usually higher than those in cabbage and Chinese cabbage fields in the same locality. Negative correlations of parasitism rates between C. plutellae and O. sokolowskii indicate a competitive relationship for host larvae between these two larval parasitoids.     

Behavioural responses of diamondback moth Plutella xylostella (Lepidoptera: Plutellidae) to extracts derived from Melia azedarach and Azadirachta indica

The impact of three different doses of botanical insecticide derived from the syringa tree, Melia azedarach and the neem tree, Azadirachta indica was tested on the behaviour of the diamondback moth, Plutella xylostella (Linnaeus). Both botanical insecticides had a significant impact on larval behaviour. At higher doses the extracts showed feeding deterrent activity, with larvae preferring the untreated sides of cabbage leaves and consuming less of the treated half of cabbage leaves. The botanical insecticides had less of an effect on the oviposition behaviour of P. xylostella moths. In laboratory and glasshouse trials, significantly fewer eggs were oviposited on the plants that had been treated with syringa extracts. Therefore, the syringa extracts appear to have a repellent effect. In contrast, when exposed to the neem extracts the moths did not discriminate between control plants and treated plants. Behavioural observation indicated that, despite the lower number of eggs oviposited on cabbage treated with syringa extracts, the moths chose cabbage treated with the highest dose of syringa more often than they chose control cabbage plants. Similar observations were found in cabbage plants treated with neem, moths chose the medium dose more often than they chose the control. Oviposition and feeding deterrent properties are important factors in pest control, and results from this study indicate that botanical insecticides have the potential to be incorporated into control programmes for P. xylostella in South Africa.     

Reproductive attributes of the eulophid Oomyzus sokolowskii,a biological control agent of diamondback moth,Plutella xylostella (Lepidoptera: Plutellidae)

Oomyzus sokolowskii (Kurdjumov) is a gregarious larval-pupal parasitoid of the diamondback moth, Plutella xylostella L. Under laboratory conditions, we explored the reproductive attributes including the functional response of O. sokolowskii and the impact of mutual interference among female parasitoids on progeny production and progeny sex ratio. In the functional response experiment, groups of 1–50 late instar larvae were exposed to groups of five female parasitoids for 48 h. Both Holling's disc equation (type II response) and Williams and Martinez's model (type III response) fit significantly better than the null-model assuming density-independent parasitation efficiency. Williams and Martinez's model was only marginally better than Holling's disc equation which explained 82% of the variation in the number of parasitized hosts. According to Holling's disc equation, the estimated maximum number of hosts parasitised during 48 h was 17 per group of five parasitoids, the estimated attack efficiency of a single female parasitoid (a) was 0.0024 larvae per hour, and the estimated handling time of a single female (T _h) was 14.4 h per host. In the mutual interference experiment 50 host larvae were exposed to different numbers of female parasitoids (P=5, 10, 20, 30, 40 and 50) for 48 h. With increasing numbers of conspecifics, the proportion of male progeny increased and total number of progeny per parasitoid decreased, yielding an estimated mutual interference constant (m) (±SE) of 0.95±0.14. The total number of wasps emerging from individual host pupae increased with increasing number of conspecifics. The age-specific fecundity of O. sokolowskii was investigated by providing 10 host larvae daily to individual female parasitoids. After 3 days of oviposition, 90% of the females were still alive and had achieved 76.4% of their lifetime fecundity. The proportion of female progeny decreased with female age. No relationship was found between female longevity and lifetime fecundity. The results suggest that parasitoid and host densities influence the progeny production and sex allocation strategy of O. sokolowskii. The information obtained from this study would help to develop a mass-rearing protocol for O. sokolowskii.