Characterization and Identification of Proteolytic Bacteria From the Gut of the Velvetbean Caterpillar (Lepidoptera: Noctuidae)

The characterization and identification of proteolytic bacteria from the gut of the velvetbean caterpillar (Anticarsia gemmatalis) were the objectives of this study. Twelve aerobic and anaerobic isolates of proteolytic bacteria were obtained from the caterpillar gut in calcium caseinate agar. The number of colony forming units (CFUs) of proteolytic bacteria was higher when the bacteria were extracted from caterpillars reared on artificial diet rather than on soybean leaves (1.73 +/- 0.35 X 10(3) and 0.55 +/- 0.22 X 10(3) CFU/mg gut, respectively). The isolated bacteria were divided into five distinct groups, according to their polymerase chain reaction restriction fragment-length polymorphism profiles. After molecular analysis, biochemical tests and fatty acid profile determination, the bacteria were identified as Bacillus subtilis, Bacillus cereus, Enterococcus gallinarum, Enterococcus mundtii, and Staphylococcus xylosus. Bacterial proteolytic activity was assessed through in vitro colorimetric assays for (general) proteases, serine proteases, and cysteine proteases. The isolated bacteria were able of hydrolyzing all tested substrates, except Staphylococcus xylosus, which did not exhibit serine protease activity. This study provides support for the hypothesis that gut proteases from velvetbean caterpillar are not exclusively secreted by the insect cells but also by their symbiotic gut bacteria. The proteolytic activity from gut symbionts of the velvetbean caterpillar is suggestive of their potential role minimizing the potentially harmful consequences of protease inhibitors from some of this insect host plants, such as soybean, with implications for the management of this insect pest species.

Avaliação da ação bioinseticida de SBTI e vicilina de Erythrina velutina em enzimas digestivas e membrana peritrófica de larvas de Plodia interpunctella (Lepidoptera: Pyralidae)

Plodia interpunctella (Indian meal moth) is a cosmopolitan pest that attacks not only a wide range of stored grain as well other food products. Due to its economic importance several researches have focused in a method with ability to control this pest with few or no damage to the environment. The study of digestive enzymes inhibitors, lectins and chitin-binding proteins, has often been proposed as an alternative to reduce insect damage. In this study we report the major classes of digestive enzymes during larval growth in P. Interpunctella, being those proteinases actives at pH 9.5 and optimum temperature of 50 oC to both larvae of the 3rd instar and pre-pupal stage of development. In vitro and zymogram assays presented the effects of several inhibitors, such as SBTI, TLCK and PMSF to intestinal homogenate of 3rd instar larvae of 62%, 92% and 87% of inhibition and In pre-pupal stage of 87%, 62 % and 55% of inhibition, respectively. Zymograms showed inhibition of two low molecular masses protein bands by TLCK and that in presence of SBTI were retarded. These results are indicative of predominance of digestive serine proteinases in gut homogenate from Plodia interpunctella larvae. This serine proteinase was then used as a target to evaluate the effect of SBTI on larvae in in vivo assay. Effect of SBTI on mortality and larval mass was not observed at until 4% of concentration (w/w) in diets. Chitin, another target to insecticidal proteins, was observed by chemical method. Moreover, optic microscopy confirmed the presence of a peritrophic membrane. Established this target, in vivo effect of EvV, a chitin binding vicilin, evaluated during the larval development of P. interpunctella and was obtained a LD50 of 0,23% and WD50 of 0,27% to this protein. Mechanism of action was proposed through of the in vivo digestibility of EvV methodology. During the passage through the larval digestive tract was observed that EvV was susceptible to digestive enzymes and a reactive fragment, visualized by Western blotting, produced by digestion was recovered after dissociation of the peritrophic membrane. The bound of EvV to peritrophic membrane was confirmed by immunohystochemical assays that showed strong immunofluorescent signal of EvV-FITC binding and peritrophic membrane. These results are a indicative that vicilins could be utilized as potential insecticide to Plodia interpunctella and a control methods using EvV as bioinsecticide should be studied to reduce lost caused by storage insect pests

Influência de genótipos de couve (Brassica oleracea L. var. acephala DC.) na biologia de Plutella xylostella (L., 1758) (Lepidoptera: Plutellidae)

Objetivou-se, com este trabalho, avaliar a influência de alguns genótipos de couve (Brassica oleracea L. var. acephala DC.) no desenvolvimento de Plutella xylostella (L., 1758) (Lepidoptera: Plutellidae). Os genótipos avaliados foram: Manteiga de Ribeirão Pires I-2620, Roxa I-919, Manteiga de São José, Manteiga de Monte Alegre, Pires 2 de Campinas, Couve Comum, Couve de Arthur Nogueira 2, Couve de Arthur Nogueira 1. Lagartas recém-eclodidas foram mantida em discos foliares de 8 cm de diâmetro para cada genótipo. Foram analisados os seguintes parâmetros: duração e viabilidade das fases larval e pupal, longevidade e fecundidade de adultos, utilizando análises paramétricas e de agrupamentos para interpretação dos dados. Observou-se um prolongamento em dias no ciclo de P. xylostella, aumento no peso de pupa e maiores valores de viabilidade e fecundidade, durante a segunda geração. O genótipo Couve de Arthur Nogueira 2 foi menos favorável ao desenvolvimento de P. xylostella nas duas gerações, e Couve Comum demonstrou maior influência negativa ao inseto na segunda geração. Manteiga de Ribeirão Pires I-2620 foi o mais suscetível nas duas gerações, agrupando com este na segunda geração Pires 2 de Campinas e Manteiga de São José.

Biology of Plutella xylostella L. (Lepidoptera: Plutellidade) Reared on Cauliflower Genotypes

This work was conducted to evaluate biological parameters of Plutella xylostella L. reared on leaves of several cauliflower genotypes under laboratory conditions. The experiment was set in a randomized block design and arranged in a 6 x 2 factorial (genotypes x generations). Leaf disks of the cultivars Barcelona, Verona, Piracicaba Precoce, Sharon, Silver Streak, and Teresopolis Gigante were placed in Petri dishes with 12 newly-hatched larvae. Leaf disks were initially changed after the fourth day, but daily afterwards until the larvae reached the pupal stage. The same procedure was adopted for the second generation. Twenty adults of each sex were separated from each genotype to evaluate their longevity, and I 0 couples from each treatment were used to assess female fecundity. The lowest larval survival was obtained on the 'Silver Streak' (78.9%) and highest on 'Verona' (97.1%). The 'Silver Streak' and `Teresopolis Gigante' showed the lowest pupal weights (4.83 mg and 5.11 mg, respectively), as well as the lowest fecundity, 119.4 and 123.0 eggs/female, respectively, while 'Piracicaba Precoce' the highest (167.7 eggs/female). Males obtained from larvae reared on `Teresopolis Gigante' and 'Silver Streak' lived shorter (5.1 days), while the short-lived females were obtained from larvae reared on 'Barcelona' and 'Verona' (4.9 and 5.0 days). Insect development was prolonged in the second generation in all tested genotypes.

Diamondback moth oviposition nonpreference in cauliflower genotypes

The Diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), is one of the main plague-insect specie of Brassicaceae plants in Brazil and all over the world. The resistant genotypes use to its control is a promising alternative. This work aimed evaluates the eggs distribution along the plant, the adults' density per plant, and determine the cauliflower genotypes effect in the P. xylostella oviposition. The experiment was carried out at FCAV/UNESP-Jaboticabal Campus Phytossanity Department (Departamento de Fitossanidade). It was evaluated the eggs distribution, the P. xylostella adults density effect using Sharon hybrid, and tests with or without choose choice to determine the P. xylostella nonpreference in the Teresopolis Gigante, Verona, Barcelona, Sharon, Silver Streak, and Piracicaba Precoce genotypes. It is possible conclude that P. xylostella has higher willingness to oviposits in the stem than in the basal leaves. The three couple density of P. xylostella per plant is the best to discriminate cauliflower genotypes regarding the resistance grade to nonpreference choose choice to oviposition. During the P. xylostella oviposition preference tests with choose choice, the genotypes Sharon, Piracicaba Precoce, Barcelona, Verona e Teresopolis Gigante are less desirable to oviposition; while during the no choose choice tests the genotypes did not differ among them.

Biologia de Plutella xylostella L. (Lepidoptera: Plutellidae) em cultivaresde couve-flor

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeito de sinigrina aplicada em folhas de brássicas sobre características biológicos de Plutella xylostella (L.) (Lepidoptera: Plutellidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Palmistichus elaeisis (Hymenoptera: Eulophidae) parasitizing pupae of Citioica anthonilis (Lepidoptera: Saturniidae) collected on Piptadenia gonoacantha (Fabaceae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Preferência de oviposição de Grapholita molesta (busck) (lepidoptera: tortricidae) em cultivares de pessegueiro

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Preferência de Apanteles galleriae Wilkinson (Hymenoptera: Braconidae) por Galleria mellonella (L.) ou Achroia grisella (Fabricius) (Lepidoptera: Pyralidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Capacidade de parasitismo de Trichogramma exiguum Pinto & Platner, 1978 (Hymenoptera: Trichogrammatidae) em ovos de Plutella xylostella (L., 1758) (Lepidoptera: Plutellidae) em diferentes temperaturas

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Biologia de Ceraeochrysa claveri Navás (Neuroptera: Chrysopidae) predando Plutella xylostella (L.) (Lepidoptera: Plutellidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Classification of crucifer cultivars based on the life-history of diamondback moth (Plutella xylostella)

We compared different cultivars and hybrids of crucifers in relation to development and life-history of diamondback moth (Plutella xylostella) to classify the plants according to their resistance to the pest. The plants used were Manteiga da Geórgia kale, Bola de Neve cauliflower, Ramoso Piracicaba Precoce broccoli, Chato-de-quintal cabbage, and the hybrid cabbages Midori, TPC668, TPC308, and TPC681. We evaluated performance daily until the pupal stage. Pupae were assessed individually to determine the pupal weight, performance, and pupal period. We determined the sex ratio, fecundity, fertility, and longevity of the emerged adults and calculated their reproductive potential. Cabbage hybrids TPC668, TPC308, and TPC681 do not support the development and reproduction of the diamondback moth. These hybrids show a level of resistance that is similar to that found the commercially available hybrid Midori and cultivar Chato de Quintal, which are known to be resistant to the diamondback moth. This finding implies that the capitata (cabbage) cultivars are the most suitable for planting because they are more resistant to pest than the cultivar's moth, acephala (kale). © 2013 Copyright Taylor and Francis Group, LLC.

Efeito de cultivares de repolho e doses de extrato aquoso de nim na alimentação e biologia de Plutella xylostella (linnaeus) (lepidoptera: Plutellidae)

The isolated and associated effects of cabbage cultivars Ruby Ball and Chato de Quintal and aqueous extract of seeds of fruits of neem (Azadirachta indica A. Juss.) were evaluated in the concentrations of 5 and 10% on Plutella xylostella (L.) attractiveness, feeding and development. In free-choice no-preference for feeding test, leaf discs of each treatment were placed in Petri dishes where three caterpillars/treatment were released while in no-choice test two caterpillars were placed in Petri dishes with one leaf disc in each dish. In the biology test, neonate larvae were transferred to Petri dishes in the proportion of one per dish, whereas the biological parameters were evaluated: larvae mortality after 1, 3 and 5 days; pupae mortality; total mortality; six day-old larvae weight; 24 hour-old pupae; and adult longevity. Cultivar Ruby Ball showed lower attractiveness to the caterpillars in free-choice test. Neem extract in both concentrations was repellent to the caterpillars in free-choice test. There were no differences in the dry mass consumed by the caterpillars in free-choice and no-choice tests. There were no differences in larval, pupal and total mortality between the cultivars. Both doses of neem extract were equally efficient in the mortality of the diamondback moth. There were no differences in caterpillars and pupae weight and in the longevity of the adults of P. xyostella between the cultivars.