Identification and recombinant expression of an antimicrobial peptide (cecropin B-like) from soybean pest Anticarsia gemmatalis

Insects can be found in numerous diverse environments, being exposed to pathogenic organisms like fungi and bacteria. Once these pathogens cross insect physical barriers, the innate immune system operates through cellular and humoral responses. Antimicrobial peptides are small molecules produced by immune signaling cascades that develop an important and generalist role in insect defenses against a variety of microorganisms. In the present work, a cecropin B-like peptide (AgCecropB) sequence was identified in the velvetbean caterpillar Anticarsia gemmatalis and cloned in a bacterial plasmid vector for further heterologous expression and antimicrobial tests. Methods AgCecropB sequence (without the signal peptide) was cloned in the plasmid vector pET-M30-MBP and expressed in the Escherichia coli BL21(DE3) expression host. Expression was induced with IPTG and a recombinant peptide was purified using two affinity chromatography steps with Histrap column. The purified peptide was submitted to high-resolution mass spectrometry (HRMS) and structural analyses. Antimicrobial tests were performed using gram-positive (Bacillus thuringiensis) and gram-negative (Burkholderia kururiensis and E. coli) bacteria. Results AgCecropB was expressed in E. coli BL21 (DE3) at 28°C with IPTG 0.5 mM. The recombinant peptide was purified and enriched after purification steps. HRMS confirmed AgCrecropB molecular mass (4.6 kDa) and circular dichroism assay showed α-helix structure in the presence of SDS. AgCrecropB inhibited almost 50% of gram-positive B. thuringiensis bacteria growth. Conclusions The first cecropin B-like peptide was described in A. gemmatalis and a recombinant peptide was expressed using a bacterial platform. Data confirmed tertiary structure as predicted for the cecropin peptide family. AgCecropB was capable to inhibit B. thuringiensis growth in vitro.(AU)

Identificación de un aislamiento argentino de Spodoptera frugiperda granulovirus / Identification of an Argentinean isolate of Spodoptera frugiperda granulovirus

Abstract The fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), is an important maize pest. Due to the environmental impact and emergence of resistance caused by chemical pesticides and transgenic events, the use of baculoviruses becomes a safe and useful alternative for its control in integrated pest management strategies. Here we report the identification of a novel isolate of a granulovirus of S. frugiperda native to the central region of Argentina, named SfGV ARG. We observed that larvae infected with SfGV ARG showed a yellowish coloration, swollen body and, in some cases, severe lesions in the last abdominal segments. We confirmed the identity of the isolate by sequencing fragments of the lef-8, lef-9 and granulin genes and by calculating evolutionary distances using the Kimura-2-Parameter model. SfGV ARG DNA restriction pattern allowed to estimate a genome of at least 135 kb.

Resumen La oruga militar tardía, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), es una plaga importante del maíz. Debido al impacto ambiental y a la aparición de resistencia causados por los pesticidas químicos y los eventos transgénicos, el uso de baculovirus resulta una alternativa útil y saludable para su control en estrategias de manejo integrado de plagas. En este trabajo reportamos la identificación de un nuevo aislamiento del granulovirus de la S. frugiperda nativo de la región central de Argentina, SfGV ARG. Se observó que larvas infectadas con SfGV ARG mostraron coloración amarillenta, hinchazón y, en algunos casos, lesiones graves en los últimos segmentos abdominales. Se confirmó la identidad del aislamiento por secuenciación de fragmentos de los genes lef-8, lef-9y granulina, y por cálculo de distancias evolutivas usando el parámetro de Kimura-2. El patrón de restricción generado con el ADN genómico de SfGV ARG permitió estimar un tamaño de genoma de al menos 135 kb.

Effects of semi-purified fractions from stems of Clusia hilariana on the development of Dysdercus peruvianus

ABSTRACT Plants represent a huge source of substances, with pharmacological potential. Brazil has a diversity of agricultural insect pests and an urgent need for safer methods of insect control. Dysdercus peruvianus (Guerin-Meneville, 1831), Pyrrhocoridae, is an economically important species of the Order Hemiptera and a pest of cotton (Gossypium hirsutum L., Malvaceae). Secondary metabolites in stems of Clusia hilariana Schltdl., Clusiaceae, such as terpenes and benzophenones, have been reported to be insecticidal. The present study investigated the effects of semi-purified fractions of hexane crude extracts from male C. hilariana stems on development of D. peruvianus. Biological parameters at different stages of development including body malformations, range of molting period and toxicity were evaluated. Most insects died and failed to develop due to attachment of their exuviae to the abdomen. Deformations of wings and defective tarsi also occurred. The secondary metabolites from semi-purified fractions of C. hilariana caused mortality, interference in molting and metamorphosis, and body deformations, probably by interacting with the neuroendocrine system. The results demonstrate the potential of C. hilariana extracts as an alternative for the control of the phytophagous insect D. peruvianus and for the development of environmentally safe and biodegradable bio insecticides.

Checklist and description of three new chromatic patterns of Pachycoris torridus (Scopoli, 1772) (Hemiptera: Scutelleridae)

In the present paper, 27 chromatic patterns of the specie Pachycoris torridus (Scopoli, 1772) were grouped and three new patterns are described. Because of this high phenotypic polymorphism, P. torridus already been registered eight times as a new specie, highlighting the importance of the application of different tools to assist in taxonomy of this hemipterous of economic importance.

No presente artigo, 27 padro˜ es croma´ ticos da espe´cie Pachycoris torridus (Scopoli, 1772) sa˜o agrupados e treˆs novos padro˜ es sa˜o descritos. Devido a esse grande polimorfismo fenoti´pico, P. torridus ja´ foi registrado oito vezes como espe´cie nova, o que ressalta a importaˆncia da aplicac¸a˜o de diferentes ferramentas para auxiliar na taxonomia desse hemi´ptero de importaˆncia econoˆmica.

Desarrollo post-embrionario, fecundidad y consumo de alimento de Dichroplus exilis (Orthoptera: Acrididae) bajo condiciones controladas

Postembryonic development, fecundity and food consumption of Dichroplus exilis (Orthoptera: Acrididae) under controlled conditions. Dichroplus exilis is a widely distributed species in Southern South America. Although there have been reports of D. exilis as an agricultural pest, some recent observations suggest that the damage attributed to D. elongatus may actually have been caused by D. exilis. This study was conducted to determine the postembryonic life cycle stages, fertility and food consumption of this species under controlled conditions (30°C, 14L-10D, 40% RH).Individuals employed belong to the laboratory-hatched first generation (F1), from adults (n=64, ♀=28, ♂=36) collected in natural grasslands near Rafaela, Santa Fe province in North- Eastern Argentina. Three cohorts of 16, 17 and 20 individuals were monitored independently in acetate tubes on a daily basis, until death of the last insect. Average fecundity was 381.84, 38.54 eggs per female. Egg-pod incubation time was 14.4, 1.08 days and six nymphal instars were recorded. Nymphal development time was 41.38, 0.71 days (I=8.73, 0.20; II=6.38, 0.24; III=5.64, 0.33; IV=7.15; 0.43; V=9.76, 0.54; IV=7.85, 0.95). The recorded food consumption was 9.89, 1.08 (mg/ind/day) for nymphs IV, 18.04, 0.73 (mg/ind/day) for nymphs V-IV, 16.76, 1.06 (mg/ind/day) for pre-reproductive males, 28.09, 1.81 (mg/ind/day) for pre-reproductive females, 7.71, 0.91 (mg/ind/day) for reproductive males and 13.06, 0.71 (mg/ind/day) for reproductive females, while the average adult food consumption, regardless of sex and reproductive status, was 16.41, 4.32mg/day. Average food consumption of adult females was 17.47, 1.15mg, and was significantly higher than that of males (10.83, 0.91mg). Data obtained in this study showed that D. exilis exhibits at least some of the biological attributes needed to configure an actual or potential agricultural pest, albeit not yet recognized as such. Field monitoring ...

La Subfamilia Melanoplinae tiene una relevancia central dentro de la acridiofauna Argentina. Varias especies suelen ser numéricamente dominantes en las comunidades de acridios del país y algunas constituyen serias plagas. El objetivo de este trabajo fue conocer el ciclo de vida postembrionario, la fecundidad y supervivencia de las hembras y el consumo de alimento de Dichruplus exilis bajo condiciones controladas (30°C; 40%HR y 14L-10O). Para ello se realizó el seguimiento de tres cohortes de 16, 17 y 20 individuos. El tiempo de incubación fue de 14.4±1.08 días. La duración del desarrollo ninfal fue 41.38; 0.71 días (I 8.73; 0.20, II 6.38; 0.24, III 5.64; 0.33, IV 7.15; 0.43, V 9.76; 0.54; IV 7.85; 0.95); la fecundidad promedio fue de 381.84, 38.54huevos/hembra. El consumo registrado fue de 9.89; 1.08 (mg/ind./día) para las ninfas de IV, 18.04; 0.73 (mg/ind./día) para las de V y IV, mientras que el consumo promedio de los adultos, sin diferenciar sexo y estado reproductivo, fue de 16.41; 4.32mg/día. Estos parámetros, al ser comparados con los de otras especies afines, indican que D. exilis exhibe atributos biológicos que condicen con los de otros melanoplinos considerados plaga.