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.     

Evidence for behavioural resistance by the diamondback moth, Plutella xylostella (L.)

Abstract:  The diamondback moth (DBM), Plutella xylostella (L.) (Lep., Plutellidae), is an important pest of brassicaceous crops worldwide. It has developed resistance to almost every synthetic insecticide applied in the field and consequently is often difficult to control in crucifer-growing areas. DBM oviposition behaviour was investigated in greenhouse and growth chamber conditions with seven cruciferous and one non-cruciferous plant varieties. In free choice tests, females deposited more eggs on the stem near the soil-stem interface than on leaves. Our findings suggest that DBM is capable of developing behavioural resistance through oviposition site selection to avoid lethal doses of foliar-applied insecticides in the field.     

Evaluating trap crops for diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae)

Potential trap crops for the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), were evaluated through a series of ovipositional preference and larval survival experiments in outdoor screenhouses in 2002 and 2003. Hosts examined as trap crops were glossy and waxy collards, Brassica oleracea L. variety acephala; Indian mustard, Brassica juncea (L.) Czern; and yellow rocket, Barbarea vulgaris (R. Br.) variety arcuata. More eggs were laid on the potential trap crops, with the exception of waxy collards, than on cabbage. When P. xylostella was offered multiple hosts at the same time, numbers of eggs laid on glossy collards, Indian mustard, and yellow rocket were 3, 18, and 12 times greater than on cabbage, respectively. Similarly, when P. xylostella was offered a single trap crop host and cabbage, numbers of eggs laid on glossy collards, Indian mustard, and yellow rocket were 300, 19, and 110 times greater than on cabbage, respectively. Our studies suggest differences in oviposition between the potential trap crops and cabbage were likely due to host volatiles, leaf morphology and color, or a combination of these factors, rather than to total leaf areas, leaf shape, or plant architecture. Two-choice tests with a Y-tube olfactometer indicated that plant volatiles were major factors in P. xylostella host preference. The percentage larval survival from egg to pupation was 22.2% on cabbage, 18.9% on waxy collards, and 24.4% on Indian mustard, whereas survival was significantly lower on glossy collards (6.7%) and yellow rocket (0%). Based on our tests, it seems that yellow rocket may be the best candidate for use as a trap crop for P. xylostella because it is highly attractive for oviposition, but larvae do not survive on it.     

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。     

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.     

小菜蛾取食对寄主植物蛋白质和糖含量影响研究

小菜蛾Plutella xylostella (L.)是重要的十字花科作物害虫,寄主植物体内营养物质的变化影响小菜蛾生长发育。本研究以小菜蛾嗜好性不同的菜心、芥菜、白菜和芥蓝等4种寄主植物为材料,应用生化方法测定了被小菜蛾取食后寄主植物体内蛋白和糖含量的变化。结果表明:小菜蛾取食2、4、6、8d后,菜心体内蛋白质含量显著升高2%～35%,葡萄糖含量明显下降9%～33%;芥菜、白菜和芥蓝体内蛋白质和葡萄糖含量显著下降1%～38%和3%～36%。小菜蛾取食后,白菜和菜心体内蔗糖含量显著升高,以菜心为害2d后升高最显著,达23.32%;芥菜和芥蓝则随为害程度的不同蔗糖含量略有差异。     

In vitro host range studies with a new baculovirus isolate from the diamondback moth Plutella xylostella (L.) (Plutellidae: Lepidoptera)

Summary The in vitro host range of a newly isolated baculovirus from the diamondback moth Plutella xylostella was tested against six lepidopteran cell lines. Two baculoviruses with host ranges from the alfalfa looper Autographa californica (A. californica multiple nucleopolyhedrovirus, AcMNPV) and the celery looper Anagrapha falcifera (AfMNPV) were also included in this study for comparative purposes. PxMNPV replicated in all six cell lines and produced occlusion bodies, with HV-AMI and TN-CLI cells producing the highest viral titers and greatest number of occlusion bodies. There was no significant replication of AcMNPV and AfMNPV in the HZ-FB33 cell line and thus no production of occlusion bodies. The restriction endonuclease profiles of the three baculoviruses showed similarities but could be readily distinguished from each other. Either HV-AM1 or TN-CL1 would be suitable cell lines for the in vitro production of PxMNPV.     

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。     

Oviposition by Plutella xylostella (Lepidoptera: Plutellidae) and effects of phylloplane waxiness

Three approaches were used to investigate effects of host plant epicuticular waxes on oviposition site selection by Plutella xylostella (L.). In the first approach, oviposition on canola (Brassica napus L.) that had epicuticular wax reduced by application of a carbamate herbicide (S-ethyl dipropylthiocarbamate) was compared with oviposition on untreated control plants. A second approach compared oviposition on sibling strains of B. napus with different wax blooms (glossy and waxy), and a third approach compared oviposition by P. xylostella on parafilm that had been applied to glossy and waxy B. napus strains for transfer of leaf components. Significantly more eggs were deposited on herbicide-treated plants (with reduced epicuticular wax) than on untreated controls. Similarly, more eggs were deposited on glossy than on waxy sibling strains of B. napus. In parafilm assays significantly more eggs were deposited on treated than on untreated parafilm. Several mechanisms could explain the differences in attractiveness of surfaces with varying wax content as oviposition sites for P. xylostella, including visual, chemical, and tactile differences between substrates. These mechanisms are discussed.     

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。     

Insecticide resistance in populations of the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), from the state of Pernambuco, Brazil

The diamondback moth Plutella xylostella (L.) has a great economic importance in Brassicaceae crops in many parts of the world. Recurrent infestations of this pest in growing areas of Pernambuco state, Brazil, have led farmers to frequently spray their crops with insecticides. However, control failures by several insecticides have been alleged by farmers. The objective of this study was to check whether resistance to insecticides could explain these control failures in P. xylostella. Populations of P. xylostella from Pernambuco were collected between January and April 2009. The resistance ratios of P. xylostella populations were compared among five different active ingredients: abamectin, methomyl, lufenuron, indoxacarb, and diafenthiuron by leaf dipping bioassays using foliar discs of kale leaves. Mortality data were submitted to probit analysis. The P. xylostella populations showed variable response and significant resistance to one or more insecticides. The population from Bezerros County exhibited the highest resistance ratios to indoxacarb (25.3 times), abamectin (61.7 times), and lufenuron (705.2 times), when compared to the reference population. The populations from Bonito and Jupi Counties were 33.0 and 12.0 times more resistant to lufenuron and abamectin, respectively, when compared with the reference population. Resistance to methomyl was the least common, but not less important, in at least four populations. These results indicated that control failures were associated with resistance by some of the evaluated insecticides, reinforcing the need for resistance management in areas of the state of Pernambuco.     

Larval survival,host plant preferences and developmental responses of the diamondback moth Plutella xylostella (Lepidoptera: Plutellidae) on wild brassicaceous species

The diamondback moth Plutella xylostella (L.) (Lepidoptera: Plutellidae) is an important pest of cultivated brassicaceous crops worldwide. The host plant preferences, developmental biology and survival and longevity of P. xylostella are relatively well understood on commercial crop species; however, its relationship with brassicaceous weeds is poorly known. Sinapis arvensis L., Erysimum cheiranthoides L. and Capsella bursa‐pastoris (L.) Medicus are among the most common brassicaceous weeds worldwide and can serve as important bridge hosts of P. xylostella. In this study, preference and performance of P. xylostella were compared on these weed species. In free‐choice situations, females deposited 5.5 and 18.8 times more eggs on S. arvensis than on E. cheiranthoides and C. bursa‐pastoris, respectively. Survival from neonate to pupa and from pupa to adult was highest on S. arvensis and E. cheiranthoides and lowest on C. bursa‐pastoris. Development was fastest, foliage consumption was greatest, pupae and silk cocoons were heaviest, adult body masses and longevities were highest and forewings were largest for both females and males when reared as larvae on S. arvensis. Realized fecundity of new generation adults was highest for individuals reared on S. arvensis compared to those reared on E. cheiranthoides or C. bursa‐pastoris. Relative growth rates of pupae and adults were highest on S. arvensis, suggesting that this plant species is a high‐quality host for P. xylostella compared with other species tested. Potential impacts of these wild brassicaceous species on P. xylostella populations are discussed.     

Seasonal population fluctuations of the diamondback moth,Plutella xylostella (L.) (Lep.: Plutellidae) on different cauliflower cultivars

The diamondback moth, Plutella xylostella (L.) (Lep.: Plutellidae) is one of the most important pests of cruciferous plants throughout the world. In recent years, it has been identified as a serious pest of the cauliflower fields in Tehran province. Resistance of P. xylostella to all main groups of insecticides has been recorded and it is ranked in the 20 most resistant pest species reported until now. According to many researchers, to solve the problem of pest resistance to chemical pesticides, an integrated pest management programme should be used. Despite this condition, it seems that the use of resistant cauliflower cultivars is an appropriate policy for integrated control of the pest in the field. In order to identify the most resistant cultivar in the field, eight cauliflower cultivars in a completely randomised design with five replicates were planted at the Shahed University research field (south of Tehran). Density of eggs, larvae and pupae of P. xylostella were measured every 10 days in these cultivars. The results showed that there is no significant difference between numbers of eggs per plant on different cultivars. But number of larvae and pupae per plant were significantly different among different cultivars. Smilla and Snow mystique cultivars had the highest number of larvae and pupae. On the other hand, Buris and Snow crown cultivars had the lowest number of pupae and Snow crown and SG cultivars had the lowest number of larvae per plant. According to the results, the Buris and Snow crown cultivars had the lowest infestation and had a kind of resistance to pest.     

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。     

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。     

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.