Effect of larval age and dosage of nuclear polyhedrosis virus on the susceptibility of diamondback moth, Plutella xylostella

A laboratory experiment was conducted to study the efficacy of nuclear polyhedrosis virus (NPV) with nine concentrations against all stadia of Plutella xylostella (L). The susceptibility of the larvae was correlated negatively with the period of development of the larvae and positively with the virus concentrations. The highest mortality of 84% was recorded in first stadium larvae compared to lowest mortality of 38% in fourth stadium larvae. The LC₅₀ was 5.5×10¹ and 4.0×10⁴ polyhedral inclusion bodies (PIB)/ml for first and fourth stadium larvae, respectively.     

Application of synthetic sex pheromone for management of diamondback moth, Plutella xylostella, in cabbage

Over a 2-year period field trials were conducted to assess the potential to disrupt mating ofPlutella xylostella (L.) using a commercial rope formulation of a 70:30 mixture of (Z)-11-hexadecenal and (Z)-11-hexadecenyl acetate, two components of the sex pheromone of the female. Screened field cages were placed into blocks of cabbage which were either treated with the pheromone or left untreated. Different densities of P. xylostella pupae were placed into each cage and then larval and pupal counts were made of the subsequent generation. In addition, sentinel females at mating stations were placed in each cage to assess the influence of the pheromone on the ability of males to locate and mate with females. Likewise, we used pheromone traps to assess whether the pheromone treatment influenced the ability of males to locate a pheromone source. In both years larval and pupal populations, produced as a result of the original inoculation, did not differ between pheromone-treated and untreated fields. The effect of pheromone treatment on larval and pupal numbers did not change with changes in inoculated P. xylostella density, however, the density of P. xylostella released caused significant differences in the density of the subsequent generation. No significant differences were detected between the number of sentinel female adult P. xylostella that successfully mated in pheromone-treated fields compared with untreated fields. Significant differences in the numbers of male P. xylostella caught in pheromone-baited traps occurred between pheromone-treated and untreated fields in the first trial of 1993, and in the first trial in 1994 but not in the second trial. Such differences are often thought of as indications of mating disruption, although our other data presented in this study and reports from other studies indicate this is not always the case. Previous studies on mating disruption of P. xylostella in larger scale field tests have been performed but the results have been variable and often ambiguous. Overall, our results indicate that mating disruption of P. xylostella with the present technology does not appear to work even under the very controlled situations which we utilized to eliminate insect movement between plots.     

A pheromone trap for dispersal of the pathogen Zoophthora radicans Brefeld. (zygomycetes: Entomophthorales) amongst populations of the diamondback moth,Plutella xylostella L. (lepidoptera: Yponomeutidae)

A sex pheromone trap designed as a novel pathogen delivery system is described. The strategy envisaged for its use is that male Plutella xylostella moths attracted to the pheromone enter the trap and are dosed with the fungus Zoophthora radicans. After habituation to the pheromone, the moths leave the trap and disperse to the crop carrying the fungal inoculum to susceptible larvae, thus initiating or enhancing an epizootic. A fluorescent marker in the trap was used to demonstrate that moths entered and exited from the trap and carried particles to crop plants at least 5 m away. As a preliminary study, this work shows the potential of this strategy for disseminating Z. radicans in populations of P. xylostella In addition, it could be considered as a model system with implications for other pest species where a pheromone and virulent pathogen are known.     

The survival ofZoophthora radicans (Zygomycetes: entomophthorales) isolates as hyphal bodies in mummified larvae ofPlutella xylostella (Lep.: Yponomeutidae)

The survival of three isolates ofZoophthora radicans (NW 250, NW 253 & NW 182) as hyphal bodies in dried larvae ofPlutella xylostella stored at 4, 10 and 20°C and 20% R.H was determined. After storage at 20°C, the production of conidia by all isolates was unaffected after 2 weeks but diminished increasingly after 4 and 8 weeks and was entirely lost after 16 weeks. By comparison conidium production at 10°C was unaffected after 16 weeks (isolates NW 250 and NW 182) and, 24 weeks (NW 253) of storage though it declined rapidly in all isolates thereafter. At 4°C many conidia were produced by all isolates even after 34 weeks of storage. These results are consistent with work on other entomophthoralean fungi in dried cadavers suggesting that this may be a common survival strategy in these fungi. NW 250, 253 and 182 were isolated fromP. xylostella in Malaysia and Taiwan, where conditions allow the host to remain active throughout the year. None produced resting sporesin vivo orin vitro but as hosts are always available the ability to survive short dry periods is probably more important than long-term survival for which resting spores are most adapted.      

Benzylideneacetone, an eicosanoid biosynthesis inhibitor enhances baculovirus pathogenicity in the diamondback moth, Plutella xylostella

Benzylideneacetone (BZA) is a monoterpenoid compound produced by an entomopathogenic bacterium, Xenorhabdus nematophila. BZA inhibits phospholipase A₂ to suppress biosynthesis of eicosanoids that mediate immune responses in insects. In response to per os infection of Autographa californica multiple nucleopolyhedrosis virus (AcMNPV), the diamondback moth, Plutella xylostella, developed red spots on the midgut epithelium. The midgut exhibiting red spot formation suffered abnormal cell integrity, such as genomic DNA fragmentation and condensed spots in the nucleoplasm. The number of red spots increased with viral dose and incubation time after the viral treatment. BZA inhibited the formation of the midgut red spots in a dose-dependent manner. However, the inhibitory effect of BZA on the red spot formation was reversed by addition of arachidonic acid, suggesting that the red spot response may be mediated by eicosanoids. BZA treatment resulted in significant enhancement of AcMNPV occlusion body (OB) pathogenicity to P. xylostella.     

Identification of chemical oviposition stimulants for the diamondback moth, Plutella xylostella, present in three species of Brassicaceae

Plant chemicals in three cruciferous crop species, Brassica napus L., B. juncea (L.) Czerniak, and Sinapis alba L., that stimulate oviposition in the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae) were investigated in laboratory bioassays. Aerial portions of 4- to 6-week-old plants were extracted and fractionated using ion-exchange liquid chromatography. The oviposition stimulants were identified as glucosinolates, which are found in all Brassicaceae species. Activity of extracts was largely eliminated by treatment with myrosinase or sulphatase, enzymes which degrade glucosinolates. Reference standards of the same glucosinolates and in the same concentrations as in the extracts were equally stimulatory. A test with eight different glucosinolates demonstrated that the moths do not discriminate between glucosinolates with different side-chain structures. However, in tests using allylglucosinolate the oviposition response was dose-dependent. One of the species tested, S. alba, contained a possible oviposition deterrent.

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Résumé Les produits chimiques trouvés dans trois espèces de crucifères cultivées, Brassica napus L., B. juncea (L.) Czerniak, et Sinapis alba L., qui stimulent l'oviposition chez la teigne des crucifères, Plutella xylostella (L.) (Lepidoptera: Plutellidae) ont été examinés. La partie aérienne des plants agés de 4 à 6 semaines a été extraite avec le methanol bouillant à 80%, le methanol pur, et l'éther. La concentration par filtration sur célite a donné comme resultat un extrait aqueux. Le materiel restant sur la célite a été dissout avec hexane pour donner un extrait lipophillique. L'extrait aqueux a été fractionné à l'aide de la chromatographie liquide sur échangeur d'ion pour donner trois fractions: neutre, cationique, et anionique. Les extraits et les fractions ont été ajustés à 1 g poids frais de tissu de plant par ml, appliqués sur du papier filtre, et exposés aux papillons femelles dans les essais de choix d'oviposition. L'oviposition a été stimulée fortement en présence de l'extrait aqueux, la fraction anionique, et quelque sous-fractions anioniques. Plus tard, il a été déterminé que ces derniers contenaient des glucosinolates.Chez les trois espèces de crucifères, les stimulants d'oviposition ont été identifiés comme étant des glucosinolates, que l'on retrouve dans toutes les espèces de crucifères. L'activité des extraits a été éliminée en grande partie par traitement avec myrosinase ou sulphatase, des enzymes qui dégradent spécifiquement les glucosinolates. Des standards de references des mêmes glucosinolates et aux même concentrations que dans les extraits ont eu également un effet stimulant. Un essai avec huit glucosinolates differentes à une concentration de 50 g/ml appliquées sur du papier filter à 3.2 g/cm² a démontré que les papillons ne discriminent pas entre les glucosinolates possédant des chaines secondaires differentes. Par contre, dans les essais utilisant l'allylglucosinolate, la réponse d'oviposition a été dépendente de la dose. S. alba a semblé contenir un inhibiteur de l'oviposition, qui est retrouvé dans l'extrait aqueux mais non pas dans la fraction anionique.

     

Reproductive compatibility and variation in survival and sex ratio between two geographic populations of Diadromus collaris, a pupal parasitoid of the diamondback moth, Plutella xylostella

Diadromus collaris (Gravenhorst)(Hymenoptera: Ichneumonidae) has been recordedin many parts of the world as a major pupalendoparasitoid of the diamondback moth Plutella xylostella (Linnaeus) (Lepidoptera:Plutellidae). Experiments were conducted tocompare reproductive compatibility andperformance as affected by temperature betweentwo geographic populations of D.collaris, one from Hangzhou, Zhejiang and theother from Lishan, Taiwan, China. The numbersand sex ratio of progeny in all possiblecrosses and backcrosses were similar to thoseobtained within each of the populations,demonstrating complete reproductivecompatibility between the two populations. Thetwo populations showed similar responses totemperature with respect to development time,adult longevity and number of host pupaeparasitized per female. However, the populationfrom Hangzhou achieved higher rates of survivalfrom larva to adult emergence at hightemperatures and had higher proportions offemale progeny at some temperatures, than thepopulation from Lishan. The Hangzhou populationachieved higher intrinsic rates of increase atall temperatures tested from 15 to33.5 °C, and the differences weresignificant at 30 and 32 °C. Thedifferences between the two populations inability to survive high temperatures mayreflect their adaptation to local climates. Thesignificance of the intraspecific variationsrevealed is discussed with regard to the use ofgeographic populations of the parasitoid in thebiological control of P. xylostella.     

Parasitism by Cotesia plutellae alters the hemocyte population and immunological function of the diamondback moth, Plutella xylostella

Cotesia plutellae, a solitary endoparasitoid wasp, parasitizes the diamondback moth, Plutella xylostella, and induces host immunosuppression and lethality in the late larval stage. This study focused on changes of cellular immunity in the parasitized P. xylostella in terms of hemocyte composition and cellular functions. In third and fourth instar larvae of nonparasitized P. xylostella, granular cells represented the main hemocyte type (60-70%) and plasmatocytes were also present at around 15% among the total hemocytes. Following parasitization by C. plutellae, the relative proportions of these two major hemocytes changed very little, but the total hemocyte counts exhibited a significant reduction. Functionally, the granular cells played a significant role in phagocytosis based on a fluorescence assay using fluorecein isothiocyanate-labeled bacteria. The phagocytic activity of the granular cells occurred as early as 5 min after incubation with the bacteria, and increased during the first 40 min of incubation. The parasitism by C. plutellae significantly inhibited phagocytosis of the granular cells. Plasmatocytes also exhibited minor phagocytic activity. Moreover, plasmatocyte phagocytosis was not inhibited by parasitism. On the other hand, hemocyte-spreading behavior in response to pathogen infection was significant only for plasmatocytes, which exhibited a characteristic spindle shape upon infection. A significant spreading of the plasmatocytes was found as early as 5 min after pathogen incubation and their ratio increased during the first 40 min.An insect cytokine, plasmatocyte-spreading peptide 1 (PSP1) from Pseudoplusia includens, was highly active in inducing plasmatocyte-spreading behavior of P. xylostella in a dose-dependent manner. P. xylostella parasitized by C. plutella was significantly inhibited in plasmatocyte-spreading in response to an active dose of PSP1. An in vivo encapsulation assay showed that the parasitized P. xylostella could not effectively form the hemocyte capsules around injected agarose beads. This research demonstrates that the parasitism of C. plutellae adversely affects the total hemocyte populations in number and function, which would contribute to host immunosuppression.     

Immunosuppression induced by entomopathogens is rescued by addition of apolipophorin III in the diamondback moth, Plutella xylostella

Apolipophorin III (ApoLpIII) has been known to play critical roles in lipid transport and immune activation in insects. This study reports a partial ApoLpIII gene cloned from the diamondback moth, Plutella xylostella. It showed that the gene was expressed in all developmental stages of P. xylostella. In larval stage, it was expressed in all tested tissues of hemocyte, fat body, gut, and epidermis. In response to bacterial challenge, the larvae showed an enhanced level of ApoLpIII expression by a quantitative real-time RT-PCR. RNA interference of ApoLpIII by its specific double stranded RNA (dsRNA) caused significant knockdown of its expression level and resulted in significant suppression in hemocyte nodule formation in response to bacterial challenge. However, larvae treated with the dsRNA exhibited a significant recovery in the cellular immune response by addition of a recombinant ApoLpIII. Parasitization by an endoparasitoid wasp, Cotesia plutellae, suppressed expression of ApoLpIII and resulted in a significant suppression in the hemocyte nodule formation. The addition of the recombinant ApoLpIII to the parasitized larvae significantly restored the hemocyte activity. Infection of an entomopathogenic bacterium, Xenorhabdus nematophila, caused potent pathogenicity of P. xylostella. However, the addition of the recombinant ApoLpIII to the infected larvae significantly prevented the lethal pathogenicity. This study suggests that ApoLpIII limits pathogenicity induced by parasitization or bacterial infection in P. xylostella.     

大菜粉蝶根虫瘟霉菌株转染小菜蛾后侵染力变化及其与寄主血淋巴酚氧化酶活性的关系

源于大菜粉蝶Pieris brassicae的根虫瘟霉Zoophthora radicans菌株R₀通过反复转染小菜蛾Plutella xylostella而分别获得转染菌株R₁、R₃和R₅。用这些菌株对小菜蛾2龄幼虫进行生物测定,发现菌株对寄主的侵染力有随转染次数增加而增强的趋势。接种后第1～6 天,R₀的LC₂₀（孢子数/mm²）分别为14.7、14.5、9.0、7.1、6.0和5.5;R₁的LC₂₀分别为9.6、5.0、4.2、3.6、3.1和3.0;R₃的LC₂₀分别为4.6、2.9、2.8、2.5、2.4和2.2; R₅的LC₂₀分别为5.2、3.7、3.2、2.8、2.6和2.6,接种后同一天菌株 R₃的LC₂₀值最小即侵染力最强。各菌株感染小菜蛾幼虫后可显著激活寄主血淋巴中的酚氧化酶活性,但R₁、R₃和R₅对酚氧化酶的激活程度显著低于原始菌株R₀。各菌株对小菜蛾的侵染力强弱指标log₁₀ (LC₂₀)与其侵染后寄主血淋巴酚氧化酶活性呈明显正相关（0.852<0.95）,表明R0在对新寄主转染过程中逐渐获得了逃避或克服新寄主免疫防御的能力,从而增强对新寄主的侵染力。     

小菜蛾碱性磷酸酶基因cDNA片段的克隆及其序列分析

利用DNAman设计简并引物,通过反转录一多聚酶链式反应(RT-PCR)克隆小菜蛾Plutella xylostella(L.)抗性、敏感种群中可能与苏云金芽孢杆菌(Bt)抗性相关的碱性磷酸酶基因。琼脂糖电泳结果显示扩增条带与预期片段长度一致,阳性克隆经测序获得了403bp的基因片断。经blast比较得出所克隆基因属于碱性磷酸酶基因。与敏感种群相比,抗性种群中该基因片段有18个碱基发生变化,有1个氨基酸发生替换(缬氨酸转变为苏氨酸)。研究结果对于进一步研究小菜蛾全基因结构、功能有重要的意义。     

抗生素和肠道细菌对小菜蛾适合度的影响

【目的】小菜蛾是危害十字花科蔬菜的世界性重要害虫。通过分析抗生素对小菜蛾的毒性效应,了解肠道细菌对小菜蛾适合度的影响,有助于更好地阐明小菜蛾肠道细菌的功能。【方法】利用抗生素处理含有高丰度肠道细菌的萝卜苗饲养品系(FZss)小菜蛾幼虫,同时利用抗生素处理饲料饲养的无肠道细菌(SLss)小菜蛾幼虫,分析抗生素及肠道细菌对小菜蛾适合度的影响。【结果】抗生素处理FZss品系小菜蛾导致了小菜蛾发育历期延长,虫重、蛹重、化蛹率、产卵量和成虫寿命降低。利用抗生素处理无肠道菌的SLss品系小菜蛾幼虫,小菜蛾化蛹率和单雌产卵量均显著降低,而对发育历期、虫重和蛹重则无影响。【结论】综合两个研究的结果发现抗生素处理后宿主适合度的降低一方面是由于抗生素的毒性效应导致,另一方面是由于小菜蛾肠道细菌的缺失引起。抗生素的毒性效应主要表现为对化蛹率和单雌产卵量的影响,而肠道细菌则对小菜蛾的发育历期、虫重、蛹重以及成虫寿命具有重要的促进作用。     

Competition and co-existence of Zoophthora radicans and Pandora blunckii, two co-occurring fungal pathogens of the diamondback moth, Plutella xylostella

The entomopathogenic fungi Zoophthora radicans and Pandora blunckii co-occur in field populations of Plutella xylostella and, therefore, are likely to interact during the infection process. We have investigated the possible outcomes of these interactions in the laboratory. Using four isolates, two of each fungal species, inter-specific interaction experiments were done in Petri dishes and on intact plants. In Petri dish experiments, larvae were inoculated directly using sporulating mats of mycelium, both species had the same opportunity to infect and only the relative concentration of conidia of each pathogen species applied was manipulated. In the intact plant experiments, larvae were placed onto fungus-contaminated plants, inoculation was passive and the probability of infection by either or both species of fungi depended on larval activity and proximity to inoculum. In the Petri dish experiment, the species with the largest concentration of conidia out-competed the other regardless of virulence, and results were similar in the intact plant experiment. The ecological implications for competition or co-existence of these two pathogens in the field are discussed.     

温度对小菜蛾生长发育和繁殖影响的研究进展

小菜蛾是典型的世界性害虫。本文综述了国内外有关温度对小菜蛾生长发育、存活、繁殖和种群增长等生物学特性影响的研究概况,提出了开展变温条件下小菜蛾生物学特性研究的重要性,这将有助于进一步弄清小菜蛾种群在田间自然条件下种群消长规律的内在机理,为利用当地的气象资料推测小菜蛾在某一时间段内的发生动态,确定防治适期,科学有效地指导防治提供理论依据。     

cDNA cloning and induction of tyrosine hydroxylase gene from the diamondback moth,Plutella xylostella

We cloned a full‐length tyrosine hydroxylase cDNA from the integument of the diamondback moth, Plutella xylostella. In the phylogenetic tree, tyrosine hydroxylase (PxTH) clustered with the other lepidopteran THs. Serine residues in the PxTH sequence, namely Ser²⁴, Ser³¹, Ser³⁵, Ser⁵³, and Ser⁶⁵, were predicted to be the target sites for phosphorylation based on PROSITE analysis. In particular, Ser³⁵ of PxTH is highly conserved across a broad phylogenetic range of animal taxa including rat and human. Western blot analysis using both PxTH‐Ab1 and PxTH‐Ab2 polyclonal antibodies verified the expression of PxTH in all life cycle stages of P. xylostella, namely the larval, pupal, and adult stages. To examine the possible immune function of PxTH in P. xylostella, PxTH gene expression was investigated by RT‐PCR and western blotting analysis after challenging P. xylostella with bacteria. PxTH expression was elevated 1 h post‐infection and was continued till 12 h of post‐infection relative to control larvae injected with sterile water. © 2010 Wiley Periodicals, Inc.     

亚精胺对小菜蛾幼虫生长及保护酶活力的影响

在实验室条件下测定经亚精胺处理后,对小菜蛾Plutella xylostella L.3龄幼虫取食量、存活、羽化率、蛹重及保护酶活力的影响。结果表明,亚精胺对小菜蛾幼虫的取食量、存活、羽化及保护酶活力均有明显的影响。经过亚精胺处理后的大部分时间段,3龄幼虫的取食量、存活率、化蛹率及体内SOD和POD活力均明显高于蒸馏水对照;对处理后24和36h CAT活力也有明显的促进作用;但亚精胺对其蛹重、蛹的羽化率却无明显的影响。     

小菜蛾幼虫偏好表达的microRNA鉴定及功能研究

[目的]microRNA(miRNA)在昆虫生长发育中发挥重要功能,本研究拟通过鉴定小菜蛾不同发育阶段的miRNA,挖掘幼虫偏好表达的miRNA及其潜在功能.[方法]对小菜蛾卵、3龄幼虫、蛹和成虫的miRNA开展高通量测序,结合生物信息学分析方法,筛选在幼虫期偏好表达的miRNA;借助实时荧光定量PCR技术,验证候选m...     

Host discrimination in two species of ichneumonid wasps, Diadegma spp., attacking larvae of Plutella xylostella

Host discrimination was investigated in two related species of ichneumonid wasps: Diadegma eucerophaga Horst. and D. fenestralis (Holmgr.) attacking a common host, the diamond-back moth Plutella xylostella (L.). In the laboratory, I studied: (1) the relative amounts of time spent between patches containing unparasitized or parasitized hosts, (2) the results of parasitoid encounters with parasitized and unparasitized hosts, (3) dissections to reveal whether or not the wasps preferred to oviposit in unparasitized hosts. D. eucerophaga showed discrimination by (1) avoiding patches containing parasitized hosts and by (2) preference to oviposit in unparasitized larvae. No discrimination was detected for D. fenestralis at any of the three levels.In the field, a population consisting primarity of D. eucerophaga was presented to plants containin either unparasitized or previously-parasitized larvae. Although superparasitism occurred, the wasps preferred to oviposit in the unparasitized larvae.

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Résumé La sélection a été examinée sur P. xylostella attaqué par deux espèces voisines Diadegma eucerophaga et D. fenestralis. La sélection a été examinée examinée à 3 niveaux au laboratoire: (1) par la comparaison des temps passés sur feuilles portant des chenilles parasitées ou non; (2) par l'analyse des résultats des rencontres avec chenilles parasitées ou non; (3) par des dissections pour examiner dans quel type la ponte a lieu de préférence. Les résultats prouvent une sélection chez D. eucerophaga qui évite les feuilles portant des chenilles parasitées, et pond de préférence dans les chenilles non parasitées. Les chenilles parasitées sont autant attaquées que les saines (insertion de la tarière), ce qui suggère peut-être un signal interne et l'aptitude à ne pas pondre après sa détection. Aucune preuve de sélection à ces trois niveaux n'a pu être apportée pour D. fenestralis. Une explication est suggérée.La sélection par une population naturelle de D. eucerophaga a aussi été étudiée dans la nature en lui présentant des chenilles parasitées ou non. Bien qu'il y ait eu un superparasitisme, les ichneumonides ont préféré pondre dans les chenilles non parasitées.

     

雷公藤生物碱对小菜蛾幼虫生长及其解毒酶系的影响

研究了雷公藤（Tripterygium wilfordii）生物碱对小菜蛾（Plutella xylostella L．）幼虫的生长发育和体内谷胱甘肽-S-转移酶（GSTs）和羧酸酯酶（CarEs）活力的影响。结果表明,在取食用LC10和LC90两种浓度雷公藤生物碱处理过的菜叶后,小菜蛾的生长发育均被抑制,抑制率随虫龄的增大而降低。2种浓度生物碱处理对GSTs酶活力的影响不相同,低浓度（LC10）处理后酶活力在1—2龄期被显著诱导升高,达对照的2．83倍,而2龄以后则被显著抑制,分别仅为对照的0．20（3龄）、0．39（4龄初）和0．27（4龄末）;在高浓度（LC90）处理下,4个不同发育阶段GSTs活力均高于对照,其中1—2龄期达对照的4．07倍,最为明显。CarEs活力在2种浓度处理下均显著高于对照,高浓度处理后在1—2、3、4龄初和4龄末期的酶活力分别为对照的2．97、3．29、3．27和2．06倍,低浓度处理后则分别为对照的2．59、2．58、2．31和1．91倍。