Global variation in the genetic and biochemical basis of diamondback moth resistance to Bacillus thuringiensis

Insecticidal proteins from the soil bacterium Bacillus thuringiensis (Bt) are becoming a cornerstone of ecologically sound pest management. However, if pests quickly adapt, the benefits of environmentally benign Bt toxins in sprays and genetically engineered crops will be short-lived. The diamondback moth (Plutella xylostella) is the first insect to evolve resistance to Bt in open-field populations. Here we report that populations from Hawaii and Pennsylvania share a genetic locus at which a recessive mutation associated with reduced toxin binding confers extremely high resistance to four Bt toxins. In contrast, resistance in a population from the Philippines shows multilocus control, a narrower spectrum, and for some Bt toxins, inheritance that is not recessive and not associated with reduced binding. The observed variation in the genetic and biochemical basis of resistance to Bt, which is unlike patterns documented for some synthetic insecticides, profoundly affects the choice of strategies for combating resistance.

Infection of the immature stages of Diadegma semiclausum, an endolarval parasitoid of the diamondback moth, by Beauveria bassiana

Interactions between the immature stages of Diadegma semiclausum, an endolarval parasitoid of Plutella xylostella, and the fungal entomopathogen Beauveria bassiana were investigated in the laboratory. Detrimental effects of B. bassiana on D. semiclausum cocoon production and adult parasitoid emergence increased with increasing pathogen concentration and some parasitoid larvae became infected by B. bassiana within hosts. The negative impact of B. bassiana on D. semiclausum cocoon production decreased as temporal separation between parasitism and pathogen exposure increased. Adult parasitoid emergence was significantly compromised by the highest rates of B. bassiana tested even when exposure of host larvae to the pathogen was delayed until one day before predicted parasitoid cocoon formation. Parasitoid pupae were infected by the pathogen in all B. bassiana treatments which did not preclude their development. (C) 2004 Elsevier Inc. All rights reserved.

Initiation of fungal epizootics in diamondback moth populations within a large field cage: proof of concept for auto-dissemination

Adult diamondback moths (DBM), Plutella xylostella L. (Lepidoptera: Plutellidae), inoculated with the fungus Zoophthora radicans, were released within a large field cage containing DBM-infested potted broccoli plants. Larvae and pupae on exposed and caged control plants were examined on five occasions over the next 48 days for evidence of Z. radicans infection. Infected larvae were first detected on exposed plants 4 days after the initial release of adults, and after 48 days the infection level reached 79%. Aerially borne conidia were a factor in transmission of the fungus. Infection had no effect on possible losses of larval and adult cadavers due to scavengers in field crops. In a trial to measure the influence of infection on dispersal, twice as many non-infected as infected males were recaptured in pheromone traps, although the difference in cumulative catch only became significant 3 days after release of the males. In a separate experiment, when adult moths were inoculated with Beauveria bassiana conidia and released into the field cage, DBM larvae collected from 37 of 96 plants sampled 4 days later subsequently died from B. bassiana infection. The distribution of plants from which the infected larvae were collected was random, but the distribution of infected larvae was clustered within the cage. These findings suggest that the auto-dissemination of fungal pathogens may be a feasible strategy for DBM control, provided that epizootics can be established and maintained when DBM population densities are low.

Contrary effects of jasmonate treatment of two closely related plant species on attraction of and oviposition by a specialist herbivore

Elevated jasmonic acid (JA) concentrations in response to herbivory can induce wounded plants to produce defences against herbivores. In laboratory and field experiments we compared the effects of exogenous JA treatment to two closely related cabbage species on the host-searching and oviposition preference of the diamondback moth (DBM), Plutella xylostella. JA-treated Chinese cabbage (Brassica campestris) was less attractive than untreated Chinese cabbage to ovipositing DBM, while JA-treatment of common cabbage (B. oleracea) made plants more attractive than untreated controls for oviposition by this insect. Similar effects were observed when plants of the two species were damaged by DBM larvae. In the absence of insect-feeding, or JA application, Chinese cabbage is much more attractive to DBM than common cabbage. Inducible resistance therefore appears to occur in a more susceptible plant and induced susceptibility appears to occur in a more resistant plant, suggesting a possible balance mechanism between constitutive and inducible defences to a specialist herbivore.

Experimental analysis of the influence of pest management practice on the efficacy of an endemic arthropod natural enemy complex of the diamondback moth.

Maximizing the contribution of endemic natural enemies to integrated pest management (IPM) programs requires a detailed knowledge of their interactions with the target pest. This experimental field study evaluated the impact of the endemic natural enemy complex of Plutella xylostella (L.) (Lepidoptera: Yponomeutidae) on pest populations in commercial cabbage crops in southeastern Queensland, Australia. Management data were used to score pest management practices at experimental sites on independent Brassica farms practicing a range of pest management strategies, and mechanical methods of natural enemy exclusion were used to assess the impact of natural enemies on introduced cohorts of P. xylostella at each site. Natural enemy impact was greatest at sites adopting IPM and least at sites practicing conventional pest management strategies. At IPM sites, the contribution of natural enemies to P. xylostella mortality permitted the cultivation of marketable crops with no yield loss but with a substantial reduction in insecticide inputs. Three species of larval parasitoids (Diadegma semiclausum Hellen [Hymenoptera: Ichneumonidae], Apanteles ippeus Nixon [Hymenoptera: Braconidae], and Oomyzus sokolowskii Kurdjumov [Hymenoptera: Eulophidae]) and one species of pupal parasitoid Diadromus collaris Gravenhorst (Hymenoptera: Ichneumonidae) attacked immature P. xylostella. The most abundant groups of predatory arthropods caught in pitfall traps were Araneae (Lycosidae) > Coleoptera (Carabidae, Coccinelidae, Staphylinidae) > Neuroptera (Chrysopidae) > Formicidae, whereas on crop foliage Araneae (Clubionidae, Oxyopidae) > Coleoptera (Coccinelidae) > Neuroptera (Chrysopidae) were most common. The abundance and diversity of natural enemies was greatest at sites that adopted IPM, correlating greater P. xylostella mortality at these sites. The efficacy of the natural enemy complex to pest mortality under different pest management regimes and appropriate strategies to optimize this important natural resource are discussed.

Oligomerization of Cry9Aa in solution without receptor binding, is not related with insecticidal activity

Background: Bacillus thuringiensis Cry toxins bind with different insect midgut proteins leading to toxin oligomerization, membrane insertion and pore formation. However, different Cry toxins had been shown to readily form high molecular weight oligomers or aggregates in solution in the absence of receptor interaction. The role of Cry oligomers formed in solution remains uncertain. The Cry9A proteins show high toxicity against different Lepidoptera, and no-cross resistance with Cry1A. Results: Cry9Aa655 protein formed oligomers easily in solution mediated by disulfide bonds, according to SDS-PAGE analysis under non-reducing and reducing conditions. However, oligomerization is not observed if Cry9Aa655 is activated with trypsin, suggesting that cysteine residues, C14 and C16, located in the N-terminal end that is processed during activation participate in this oligomerization. To determine the role of these residues on oligomerization and in toxicity single and double alanine substitution were constructed. In contrast to single C14A and C16A mutants, the double C14A–C16A mutant did not form oligomers in solution. Toxicity assays against Plutella xylostella showed that the C14A–C16A mutant had a similar insecticidal activity as the Cry9Aa655 protein indicating the oligomers of Cry9Aa formed in solution in the absence of receptor binding are not related with toxicity. Conclusions: The aggregation of Cry9Aa655 polypeptides was mediated by disulfide bonds. Cry9Aa655 C14 and C16C are involved in oligomerization in solution. These aggregate forms are not related to the mode of action of Cry9Aa leading to toxicity.

Efectividad de cuatro insecticidas en el control de larvas de Plutella maculipennis (Curtis) y Leptophobia aripa (Boisd) en el cultivo del repollo (Brassica oleracea var. superette)

Las plagas de foliadores como plutella y leptophobia constituyen uno de los principales problemas fitosanitarios en el cultivo del repollo en Nicaragua. Fundamentalmente contra plutella se han aplicado de forma irracional diversos insecticidas; solos y en combinaciones, lo que posiblemente ha hecho que esta plaga desarrolle resistencia a algunos productos plaguicidas, En este estudio se evaluo la efectividad de cuatro insecticidas: Dipel Decis, Lannate y Tamaron , contra las larvas de estos dos lepidópteros; encontrándose que estos tienen buen control sobre Leptophobia: sin embargo, contra Plutella el único efectivo fue Dipel, resultando los productos químicos en efectivo contra ella, esa diferencia de efectividad podría estar asociada al factos resistencia que plutella podría tener hacia dichos productos; a pesar de estos resultados los productores continúan utilizando estos insecticidas debido al bajo costo de los mismos. Producto de la política de subsidio del estado para los agroquímicos y al alto precio que reciben por el repollo debido a la inflación; lo que genera un desequilibrio entre el valor económico del repollo y los costos de aplicación de los insecticidas lo que les permite tener siempre ganancias aunque realicen numerosas aplicaciones teniendo esto como resultado una alteración en el ecosistema, por tanto , se debe discutir ampliamente el impacto que tiene la policía de subsidio de los agroquímicos en la agricultura nacional. El ingreso económico en repollo es una función del número de cabezas formadas y el precio /cabeza; leptophobia influye el rendimiento, afectando el número de cabezas formadas; el precio/cabezas esata afectado por las poblaciones de plutella y leptophobia por lo que la efectividad de los insecticidas se evalúa en base al ingreso económico que combina el rendimiento y la calidad del repollo en relación con la población de plagas que permaneces en el cultivo.

Evaluación de productos botánicos y biológicos para el control de plagas defoliadoras en el cultivo del repollo (Brassica oleracea) híbrido Izalco

Durante el ciclo del cultivo se presentaron plagas como: P. xylostella, Spodoptera sp. y Helulla phydeliales. Presentando mayor incidencia P. Jlylastello con un comportamiento diferente en cada etapa del cultivo en contrando que en las etapas de formación y llenado de cabezas (30-50 y 50-80 DDT) se presentaron las mayores poblaciones, considerando estas dos etapas corno períodos críticos, ya que la plaga en esta etapa afecta la calidad del producto lo que influye económicamente. Durante el ciclo del cultivo fue notorio el control eficaz que ejercio el tratamiento Extracto Acuoso de Semilla de Neem sobre P. xylostella; seguido por los tratamientos javelin (B. thuringiensis) y júpiter (Chlorfluasuron) que lograron mantener las poblaciones bajas, no así los tratamientos Dipel comercial, BT. Nacional y Extracto Acuoso de Semilla de Mamey los que mantuvieron poblaciones similares a las presentadas por el testigo (sin aplicación). El análisis económico de los tratamientos, indicó que el tratamiento de mayor rentabilidad fue el Extracto Acuoso de Semilla de Neem seguido por el tratamiento júpiter, los cuales poseen una tasa de retorno marginal aceptable en cambio los tratamientos, Dipel Comercial, Extracto Acuoso de Semilla de Mamey, BT Nacional no son recomendables para el control de P. xylostella.

Efecto de policultivo repollo-tomate sobre la entomofauna del cultivo de repollo

En el estudio realizado en el Valle de Sebaco, durante el periodo de Nov.90 - Mar.91, establecido con el objetivo de determinar el efecto del policultivo: (Repollo Tomate) sobre la entomofauna del cultivo de repollo (Brassica oleracea L.), sobre la calidad y rendimiento del repollo, cuyos tratamientos comprendieron monocultivo (Repollo­ Repollo) y policultivo (Repollo-Tomate) en dos diferentes modalidades de siembra (Tiempo: 10, 30 Dias Antes del Transplante, y espacio: surco, cama). Las poblaciones de P.xylostella fueron menores en policultivo, sin embargo sobre el resto de poblaciones de insectos (Diabrotica sp., áfidos, y Creontiades sp.) este sistema de asocio no tuvo efecto reductor. Ofreciendo igual protección de este defoliador al cultivo de repollo en las dos modalidades de siembra en estudio (tiempo y espacio). En la incidencia de insectos benéficos como: Arañas, D.insulare fueron iguales, en ambos sistemas de siembra, solo las poblaciones de Polybia sp. fueron Superiores en monocultivo, sin embargo al establecer una relación de insecto plagas (P. xyylostella) por insecto benéfico estos fueron superiores en policultivo lo que permitió ser un mecanismo de control de P, xylostella en el cultivo de repollo. de igual manera el porcentaje de parasitismo de D. insular e fue de 31.87% en monocultivo y de 34.72% para poliCultivo permitiendo este sistema de siembra un aumento en los enemigos naturales los que son un mecanismo de control de P. xylostella. El cultivo de tomate ejerce protección al cultivo de repollo de las plagas defoliadoras lo que permite obtener• una mejor calidad de cabezas con menor cantidad de área foliar dañada siendo del 20%, un porcentaje de cabezas formadas de 74% y peso de 1.10kg. lo que permite incrementar los rendimientos del cultivo de repollo. El Cultivo de tomate en sus dos modalidades de siembra permite obtener similares ingresos, y permite incrementar los rendimiento en el sistema de siembra en policultivo.

Efecto de poli-cultivo (repollo-tomate; repollo-zanahoria) sobre la incidencia de defoliadores del cultivo de repollo (Brassica oleracea) var. Supperette

Se estableció un experimento de campo con el objetivo de evaluar el efecto de los poli-cultivos repollo-tomate, repollo-zanahoria sobre la incidencia de las principales plagas defoliadoras del cultivo de repollo brassica oleracea además se evaluó un tratamiento de repollo en mono-cultivo donde se realizó aplicación del insecticida biológico dipel ( bacillus thuringensis. Las plagas predominantes durante el ensayo fueron plutella xilostella helulla phidilealis y ascia monuste. Para plutella se encontró diferencia significativa entre los tratamiento, siendo el poli-cultivo repollo-zanahoria que presento las menores poblaciones, en cuanto a helulla no se observó un efecto beneficioso del poli-cultivo ya que las poblaciones se mantuvieron estadísticamente iguales en los cuatro tratamientos, igual sucedió con las poblaciones de ascia. Se presentó mayores ingreso brutos en las parcelas de poli-cultivo repollo-tomate y en el mono-cultivo con aplicación de dipel, en el poli-cultivo repollo-zanahoria y en el mono-cultivo de repollo sin aplicaciones de insecticida los ingresos fueron bajos

Effects of a sex-ratio distorting endosymbiont on mtDNA variation in a global insect pest

Background: Patterns of mtDNA variation within a species reflect long-term population structure, but may also be influenced by maternally inherited endosymbionts, such as Wolbachia. These bacteria often alter host reproductive biology and can drive particular mtDNA haplotypes through populations. We investigated the impacts of Wolbachia infection and geography on mtDNA variation in the diamondback moth, a major global pest whose geographic distribution reflects both natural processes and transport via human agricultural activities. Results: The mtDNA phylogeny of 95 individuals sampled from 10 countries on four continents revealed two major clades. One contained only Wolbachia-infected individuals from Malaysia and Kenya, while the other contained only uninfected individuals, from all countries including Malaysia and Kenya. Within the uninfected group was a further clade containing all individuals from Australasia and displaying very limited sequence variation. In contrast, a biparental nuclear gene phylogeny did not have infected and uninfected clades, supporting the notion that maternally-inherited Wolbachia are responsible for the mtDNA pattern. Only about 5% (15/306) of our global sample of individuals was infected with the plutWBI isolate and even within infected local populations, many insects were uninfected. Comparisons of infected and uninfected isofemale lines revealed that plutWBI is associated with sex ratio distortion. Uninfected lines have a 1:1 sex ratio, while infected ones show a 2:1 female bias. Conclusion: The main correlate of mtDNA variation in P. xylostella is presence or absence of the plutWBI infection. This is associated with substantial sex ratio distortion and the underlying mechanisms deserve further study. In contrast, geographic origin is a poor predictor of moth mtDNA sequences, reflecting human activity in moving the insects around the globe. The exception is a clade of Australasian individuals, which may reflect a bottleneck during their recent introduction to this region.