Search

The South American soybean pest, Rachiplusia nu (Guenée), is naturally infected by Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and Rachiplusia nu nucleopolyhedrovirus (RanuNPV). We compared their pathogenicity to fourth-instar R. nu larvae, by evaluating time to death and virus spread throughout the tissues in single and mixed infections. Bioassays showed that generalist AcMNPV had a faster speed of kill than specific RanuNPV, while the mixed-virus treatment did not statistically differ from AcMNPV alone. Histopathology evidenced similar tissue tropism for both viruses, but co-inoculation resulted in mostly AcMNPV-infected cells. In sequential inoculations, however, the first virus administered predominated over the second one. Implications on baculovirus interactions and biocontrol potential are discussed. Instituto de Microbiología y Zoología Agrícola (IMYZA) Fil: Decker Franco, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Decker Franco, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Taibo, Catalina Beatriz. Instituto Nacional de Tecnología Agropecuaria (INTA). Centro de Investigación en Ciencias Veterinarias y Agronómicas. Laboratorio Integral de Microscopía; Argentina Fil: Di Rienzo, Julio A. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias. Cátedra de Estadística y Biometría; Argentina Fil: Alfonso, Victoria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Alfonso, Victoria. Instituto Nacional de Tecnología Agropecuaria (INTA). Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina Fil: Arneodo, Joel D. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Arneodo, Joel D. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina

Chaetosiphon fragaefolii Cockerell (Hemiptera: Aphididae) is a worldwide strawberry pest with scarce records of parasitoids for its control. The parasitization rate of two generalist parasitoids, Aphidius colemani Viereck and Aphidius matricariae Haliday, their behavior and aphid defensive behaviors were evaluated with Aphis gossypii Glover (Hemiptera: Aphididae) as a comparative host. Defensive endosymbionts were also surveyed. C. fragaefolii was never parasitized in simple-choice tests, whereas in preference tests both parasitoids chose A. gossypii over C. fragaefolii. No contacts of A. matricariae with C. fragaefolii were observed while A. colemani made more antennal contacts and “stings” on A. gossypii than on C. fragaefolii with the latter exhibiting less defensive behaviors against the parasitoid. The bacterium Acinetobacter sp. was detected in all samples of C. fragaefolii, but with an unknown function. Further research on the possible defensive role of the glandular hairs covering the body of this aphid, the role of chemical signals and the existence of defensive internal mechanisms against parasitoids could provide plausible explanations for the absence of parasitism found.

The termite gut microbiome is dominated by lignocellulose degrading microorganisms. This study describes the intestinal microbiota of four Argentinian higher termite species with different feeding habits: Microcerotermes strunckii (hardwood), Nasutitermes corniger (softwood), Termes riograndensis (soil organic matter/grass) and Cornitermes cumulans (grass) by deep sequencing of amplified 16S rRNA and ITS genes. In addition, we have performed a taxonomic and gut community structure comparison incorporating into the analysis the previously reported microbiomes of additional termite species with varied diets. The bacterial phylum Spirochaetes was dominant in the guts of M. strunckii, N. corniger and C. cumulans, whereas Firmicutes predominated in the T. riograndensis gut microbiome. A single bacterial genus, Treponema (Spirochaetes), was dominant in all termite species, except for T. riograndensis. Both in our own sequenced samples and in the broader comparison, prokaryotic α-diversity was higher in the soil/grass feeders than in the wood feeders. Meanwhile, the β-diversity of prokaryotes and fungi was highly dissimilar among strict wood-feeders, whereas that of soil- and grass-feeders grouped more closely. Ascomycota and Basidiomycota were the only fungal phyla that could be identified in all gut samples, because of the lack of reference sequences in public databases. In summary, higher microbial diversity was recorded in termites with more versatile feeding sources, providing further evidence that diet, along with other factors (e.g., host taxonomy), influences the microbial community assembly in the termite gut. Instituto de Biotecnología Fil: Vikram, Surendra. University of Pretoria. Centre for Microbial Ecology and Genomics. Department Biochemistry. Genetics and Microbiology; Sudáfrica Fil: Arneodo Larochette, Joel Demian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Arneodo Larochette, Joel Demian. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Calcagno, Javier. Universidad Maimonides. Centro de Ciencias Naturales, Ambientales y Antropológicas; Argentina Fil: Ortiz, Maximiliano. University of Pretoria. Centre for Microbial Ecology and Genomics. Department Biochemistry. Genetics and Microbiology; Sudáfrica Fil: Mon, Maria Laura. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Mon, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Etcheverry, Clara. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas y Naturales y Agrimensura. Biología de los Invertebrados; Argentina Fil: Cowan, Donald. University of Pretoria. Centre for Microbial Ecology and Genomics. Department Biochemistry. Genetics and Microbiology; Sudáfrica Fil: Talia, Paola Mónica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Talia, Paola Mónica. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

In this study, we used shotgun metagenomic sequencing to characterise the microbial metabolic potential for lignocellulose transformation in the gut of two colonies of Argentine higher termite species with different feeding habits, Cortaritermes fulviceps and Nasutitermes aquilinus. Our goal was to assess the microbial community compositions and metabolic capacity, and to identify genes involved in lignocellulose degradation. Individuals from both termite species contained the same five dominant bacterial phyla (Spirochaetes, Firmicutes, Proteobacteria, Fibrobacteres and Bacteroidetes) although with different relative abundances. However, detected functional capacity varied, with C. fulviceps (a grass-wood-feeder) gut microbiome samples containing more genes related to amino acid metabolism, whereas N. aquilinus (a wood-feeder) gut microbiome samples were enriched in genes involved in carbohydrate metabolism and cellulose degradation. The C. fulviceps gut microbiome was enriched specifically in genes coding for debranching- and oligosaccharide-degrading enzymes. These findings suggest an association between the primary food source and the predicted categories of the enzymes present in the gut microbiomes of each species. To further investigate the termite microbiomes as sources of biotechnologically relevant glycosyl hydrolases, a putative GH10 endo-β-1,4-xylanase, Xyl10E, was cloned and expressed in Escherichia coli. Functional analysis of the recombinant metagenome-derived enzyme showed high specificity towards beechwood xylan (288.1 IU/mg), with the optimum activity at 50 °C and a pH-activity range from 5 to 10. These characteristics suggest that Xy110E may be a promising candidate for further development in lignocellulose deconstruction applications. Fil: Romero Victorica, Matias. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Soria, Marcelo Abel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; Argentina Fil: Batista García, Ramón Alberto. Universidad Autónoma del Estado de Morelos.; México Fil: Ceja Navarro, Javier A.. Lawrence Berkeley National Laboratory; Estados Unidos Fil: Vikram, Surendra. University of the Witwatersrand; Sudáfrica Fil: Ortiz, Maximiliano. University of Pretoria; Sudáfrica Fil: Ontañon, Ornella Mailén. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Ghio, Silvina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Martínez Ávila, Liliana. Universidad Autónoma del Estado de Morelos.; México Fil: Quintero García, Omar Jasiel. Universidad Autónoma del Estado de Morelos.; México Fil: Etcheverry, Clara. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Biología. Cátedra Biología de los Invertebrados; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Campos, Eleonora. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Cowan, Donald Arthur. University of Pretoria; Sudáfrica Fil: Arneodo Larochette, Joel Demián. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Talia, Paola Monica. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina

Laboratory trials were performed to determine the impact of three Neotropical predatory coccinellids (Cycloneda sanguinea, Eriopis connexa and Coleomegilla quadrifasciata) and a minute pirate bug (Orius insidiosus) on Chaetosiphon fragaefolii, an important strawberry aphid pest. The predation on C. fragaefolii nymphs and adults, as well as the time to the first attack of all predators were compared with predation on Aphis gossypii. Predator preferences for prey and aphid defensive behavior were also evaluated. Moreover, the effect of coccinellids on C. fragaefolii population growth was assessed in experimental greenhouse conditions. The predation rate varied among predators, being significantly lower for O. insidiosus than for the coccinellids. Consumption was higher on A. gossypii than on C. fragaefolii, regardless of the aphids developmental stage. The time to the first attack of all predators was longer in the presence of C. fragaefolii. Walking away and cornicle secretion were the most common antipredator behaviors of aphid against coccinellids and O. insidiosus, respectively. Coccinellids preferred A. gossypii over C. fragaefolii, while O. insidiosus showed indifference. Cycloneda sanguinea and E. connexa exhibited the highest suppression effect on the growth rate of C. fragaefolii. Thus, the four predators evaluated could contribute to reduce strawberry aphid populations, especially C. sanguinea and E. connexa. Fil: Francesena, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; Argentina Fil: Rocca, Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; Argentina Fil: Rizzo, Maria Estefania. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; Argentina Fil: Arneodo Larochette, Joel Demián. Instituto Nacional de Tecnología Agropecuaria; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Greco, Nancy Mabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; Argentina. Comisión de Investigaciones Científicas de la Provincia de Buenos Aires; Argentina

The Autographa californica multiple nucleopolyhedrovirus (AcMNPV) ac12 gene, which is conserved in ten other baculovirus, codes a predicted 217 amino acid protein of unknown function. In this study, we investigated the role of ac12 during baculovirus infection, by generating an ac12 knockout virus. The transfection of the recombinant genome in insect cells resulted in unaltered viral dispersion and occlusion body production when compared to the control bacmid. This finding demonstrates that ac12 is a non-essential gene. Transmission and scanning electron microscopy (SEM) analyses showed that ac12 knockout virus produced occlusion bodies morphologically similar to those obtained with the control and capable to occlude virions. However, a slight but significant size difference was detected by SEM observation of purified occlusion bodies. This difference suggests that ac12 may be involved in regulatory pathways of polyhedrin production or occlusion body assembly without affecting either viral occlusion or oral infectivity in Rachiplusia nu larvae. This was evidenced by bioassays that showed no significant differences in the conditions tested. A qPCR analysis of viral gene expression during infection evidenced regulatory effects of ac12 over some representative genes of different stages of the viral cycle. In this study, we also showed that ac12 is transcribed at early times after infection and remains detectable up to 72 hours post-infection. The mRNA is translated during the infection and results in a protein that encodes an F-box domain that interacts in vivo and in vitro with S phase kinase associated protein 1 (SKP1) adaptor protein, which is potentially involved in protein ubiquitination pathways.

Most of our knowledge about the consequences of tropospheric ozone pollution on aphid-plant interaction assumes the absence of a direct effect of ozone on aphids. However, the biological effects of ozone encompass changes in reactive oxygen species signalling, oxidative stress accumulation, and the immune response of both plants and animals. The main objective of this work was to evaluate the direct effect of ozone on Metopolophium dirhodum (Walker) aphids (Hemiptera: Aphididae), linking mechanisms that operate at individual scale (oxidative stress parameters and aphid-bacteria symbiosis persistence) with their consequences at population scale (mortality, dispersion, and population growth). Two experiments were conducted in open-top chambers with three contrasting ozone exposure regimes (6 h at 0.5 ± 0.3, 50 ± 5, or 150 ± 10 p.p.b.) and artificial diets to evaluate the direct effect of ozone on aphids. Exposure of aphids to ozone increased insect mortality compared to the control treatment. However, the populations established from the surviving insects of each treatment increased similarly. Consistently, aphid symbiosis with Buchnera aphidicola Munson et al. (Proteobacteria) and Hamiltonella defensa Moran et al. (Enterobacteriaceae) persisted in all treatments. Ozone also affected aphid behaviour. The proportion of insects dispersing from diet cages was lower in the ozone treatments than in the control treatment. In addition, lipid peroxidation was higher at 150 p.p.b. than at 50 p.p.b. treatment, although not different from that in the control. The reduction in aphid dispersion coupled with increased mortality suggests that direct exposure to ozone could lower aphid efficiency as virus vectors. These results could be partially associated with mechanisms operating at individual scale (accumulation of oxidative damage). Overall, these experiments encourage reconsidering the impact of the direct effects of ozone on aphids when assessing the consequences of this component of global change on plant-aphid interactions. Fil: Telesnicki, Marta Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; Argentina Fil: Martinez-Ghersa, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; Argentina Fil: Arneodo Larochette, Joel Demián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Ghersa, Claudio Marco. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; Argentina

The Helicoverpa/Heliothis complex can cause serious damage to agricultural crops. Phenotypic similarity makes it difficult to discriminate between closely related Helicoverpa species. Currently, morphology of the male genitalia complemented with molecular techniques constitutes the best approach for species identification. In this work, a broad microscopic examination of adult Helicoverpa zea (Boddie) males (n¼200) captured in central Argentina was carried out in order to provide a detailed description of the valvae and the phallus. A considerable degree of variability was recorded. Both rounded and sharp valve apices were observed and valvae were not always parallel-sided. Most evident differences were detected concerning the number of cornuti on the phallus. A range of 15–21 cornuti per phallus was recorded, the mode being 18. A significant minority of the samples (3.5%) displayed an abnormal genital condition showing a constricted phallus lacking cornuti, and pointed valvae. This form was initially attributed to a distinct species, Heliothis stombleri, and later proposed as a synonym of H. zea based on additional morphological observations and molecular studies. Here, a phylogenetic analysis combining mitochondrial (cytochrome oxidase subunit I) and nuclear (elongation factor –1 alpha) genes was performed on these and other Helicoverpa specimens collected in the same geographical region, in order to further verify the taxonomic status of H. stombleri. The tree topology clearly grouped H. stombleri with H. zea, supporting the assumption that the former represents, in fact, an anomalous form of the latter. Further experiments are needed to clarify the etiology of this anomaly and its persistence over time. El complejo Helicoverpa/Heliothis produce graves daños a la agricultura. Las especies del género Helicoverpa son difíciles de diferenciar porque poseen un fenotipo similar. Actualmente, su identificación se basa en características morfológicas de la genitalia masculina y en el uso de marcadores moleculares. En este trabajo, se examinaron por microscopía machos adultos de Helicoverpa zea (Boddie; n = 200) capturados en la zona central de Argentina, a fin de detallar la forma y variabilidad de valvas y falo. Las valvas no siempre presentaron lados paralelos, y mostraron ápices agudos o redondeados. Las diferencias más evidentes se refirieron al número de cornuti del falo (15 a 21, moda = 18). Una minoría significativa de las muestras (3,5%) exhibió una fuerte anomalía en el aspecto del aparato reproductor: falo sin cornuti, estrechado en su extremo, y valvas terminadas en punta. Esta variante, que había sido atribuida inicialmente a una especie distinta, Heliothis stombleri, fue luego considerada como sinónimo de H. zea mediante estudios morfológicos y moleculares. Para verificar esta conclusión, se llevó a cabo un análisis filogenético que combinó genes mitocondriales (COI) y nucleares (EF-1α) a partir de especímenes de H. stombleri y otras Helicoverpa spp. provenientes de la misma región. La topología de los árboles agrupó claramente a H. stombleri con H. zea, permitiendo suponer que, en efecto, ambas formas constituyen la misma especie. Experimentos adicionales son necesarios para clarificar la etiología de esta anomalía, así como su persistencia en el tiempo. Instituto de Biotecnología Fil: Balbi, Emilia Ines. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juárez; Argentina Fil: Flores, Fernando Miguel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juárez; Argentina Fil: Tosto, Daniela Sandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Arneodo Larochette, Joel Demian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Microbiología y Zoología Agrícola; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Typical baculovirus infection symptoms were observed in Chrysodeixis includens Walker (Lepidoptera: Noctuidae) larvae from a laboratory rearing in Tucumán, Argentina. Scanning electron microscopy revealed the occurrence of polyhedral occlusion bodies displaying a heterogeneous morphology. Amplification, sequencing, and phylogenetic analysis of conserved polyhedrin, lef-8 and lef-9 genes identified the pathogen, for the first time in Argentina, as a variant of Chrysodeixis includens nucleopolyhedrovirus (ChinNPV). The biocontrol potential of this new isolate is worthy of future research. Se observaron síntomas típicos de infección por baculovirus en larvas de Chrysodeixis includens Walker (Lepidoptera: Noctuidae) criadas en laboratorio en Tucumán, Argentina. Mediante microscopía electrónica de barrido, se observaron cuerpos de oclusión poliédricos de morfología heterogénea. La amplificación, secuenciación y análisis filogenético de tres genes conservados (poliedrina, lef-8 y lef-9) permitieron identificar al patógeno, por primera vez en Agentina, como una variante de Chrysodeixis includens nucleopolyhedrovirus (ChinNPV). Se prevé investigar el potencial biocontrolador de este nuevo aislamiento. Instituto de Microbiología y Zoología Agrícola Fil: Arneodo Larochette, Joel Demian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Microbiología y Zoología Agrícola; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Dami, Luciana Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Tecnología Agroindustrial del Noroeste Argentino. Provincia de Tucumán. Ministerio de Desarrollo Productivo. Estación Experimental Agroindustrial ; Argentina Fil: Jakubowicz, Violeta. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Microbiología y Zoología Agrícola; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Alzogaray, Raúl Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones Científicas y Técnicas para la Defensa. Centro de Investigación de Plagas e Insecticidas; Argentina Fil: Taibo, Catalina Beatriz. Instituto Nacional de Tecnología Agropecuaria (INTA). Centro de Investigación en Ciencias Veterinarias y Agronómicas. Servicio de Microscopía; Argentina

Rachiplusia nu (Lepidoptera: Noctuidae) is a key soybean pest in Argentina. Current management of this moth relies mainly on the use of synthetic insecticides and transgenic plants. In search of biological control-based alternatives, a baculovirus from R. nu (hereafter RanuNPV) was characterized and its insecticidal properties tested under laboratory conditions. RanuNPV occlusion bodies (OBs) were nearly tetrahedral, averaging 1.0 ± 0.2 µm in their longest edge and containing singly enveloped nucleocapsids. Histopathology of infected late-instar larvae revealed broad tissue tropism, where fat bodies and epidermis were the most affected organs. Phylogenetic analysis of concatenated polh, lef-8 and lef-9 partial sequences classified RanuNPV as a new species that clusters with other group II alphabaculoviruses infecting larvae of Plusiinae. Bioassays performed with R. nu neonates determined the median lethal dosage to be approximately 2.5 OBs/larva; most insects died within 4–5 days post inoculation showing typical baculovirus-induced liquefaction. No effects were observed in other lepidopteran species assayed, including Spodoptera frugiperda, Cydia pomonella and Diatraea saccharalis. High pathogenicity and host specificity make RanuNPV a good candidate for controlling R. nu.

Entomopathogens help regulate natural insect populations, and their use in pest management is the subject of active research. Classical and molecular techniques are currently available to elucidate disease etiology and pathogenesis. Here, we performed an Azan trichrome stain for light microscopic visualization of pathogens in the larval tissues of two Neotropical lepidopterans. Epinotia aporema larvae exhibiting symptoms of microsporidiosis were fortuitously detected in a rearing facility (IMyZA-INTA, Argentina), while Helicoverpa gelotopoeon specimens were experimentally infected with an Argentinean isolate of Helicoverpa armigera nucleopolyhedrovirus (HearNPV). Caterpillars were fixed in Duboscq-Brasil and paraffin embedded. Microtome sections (5 μm-thick) were deparaffinized and stained2 for observation under a Nikon Eclipse 80i microscope equipped with a Nikon DS-U3 digital camera. Inst. de Microbiología y Zoología Agrícola IMyZA Fil: Arneodo Larochette, Joel Demian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Microbiología y Zoología Agrícola; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Taibo, Catalina Beatriz. Instituto Nacional de Tecnología Agropecuaria (INTA). Centro de Investigación en Ciencias Veterinarias y Agronómicas. Servicio de Microscopía; Argentina Fil: Sciocco, Alicia Ines. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Microbiología y Zoología Agrícola; Argentina

Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae: Heliothinae) is among the most voracious global pests of agriculture. Adults of this species were identified recently in northern Argentina by dissection of male genitalia. In this work, a rapid and simple molecular tool was designed to distinguish H. armigera from the morphologically similar indigenous bollworms Helicoverpa zea (Boddie) and Helicoverpa gelotopoeon (Dyar), regardless of the life stage. Amplification of partial COI gene with a new primer pair, and subsequent digestion with endonuclease HinfI, yielded different RFLP profiles for the three main Helicoverpa pests currently present in South America. The method was validated in Helicoverpa specimens collected across Argentina, whose identity was further corroborated by COI sequencing and phylogenetic analysis. The data reported here constitute the first molecular confirmation of thispest in the country. The survey revealed the occurrence of H. armigera in northern and central Argentina, including the main soybean- and maize-producing area. Fil: Arneodo Larochette, Joel Demián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Balbi, Emilia I.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Córdoba. Estación Experimental Agropecuaria Marcos Juárez; Argentina Fil: Flores, Fernando M.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Córdoba. Estación Experimental Agropecuaria Marcos Juárez; Argentina Fil: Sciocco Cap, Alicia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina

Saccharum officinarum bagasse (common name: sugarcane bagasse) and Pennisetum purpureum (also known as Napier grass) are among the most promising feedstocks for bioethanol production in Argentina and Brazil. In this study, both biomasses were assessed before and after acid pretreatment and following hydrolysis with Nasutitermes aquilinus andCortaritermes fulviceps termite gut digestome. The chemical composition analysis of the biomasses after diluted acid pretreatment showed that the hemicellulose fraction was partially removed. The (hemi) cellulolytic activities were evaluated in bacterial culture supernatantsof termite gut homogenates grown in treated and untreated biomasses. In all cases, we detected significantly higher endoglucanase and xylanase activities using pretreated biomasses compared to untreated biomasses, carboxymethylcellulose and xylan. Several protein bands with (hemi) cellulolytic activity were detected in zymograms and two-dimensionalgel electrophoresis. Some proteins of these bands or spots were identified as xylanolytic peptides by mass spectrometry. Finally, the diversity of cultured cellulolytic bacterial endosymbionts associated to both Argentinean native termite species was analyzed. This study describes, for the first time, bacterial endosymbionts and endogenous (hemi) cellulases of two Argentinean native termites as well as their potential application in degradation of lignocellulosic biomass for bioethanol production. Fil: Ben Guerrero, Emiliano. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina Fil: Arneodo Larochette, Joel Demián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Bombarda Campanha, Raquel. Ministerio da Agricultura Pecuaria e Abastecimento de Brasil. Empresa Brasileira de Pesquisa Agropecuaria; Brasil Fil: Oliveira, Patrícia Abrão de. Ministerio da Agricultura Pecuaria e Abastecimento de Brasil. Empresa Brasileira de Pesquisa Agropecuaria; Brasil Fil: Labate, Mônica T. Veneziano. Universidade de Sao Paulo; Brasil Fil: Cataldi, Thaís Regiani. Universidade de Sao Paulo; Brasil Fil: Campos, Eleonora. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Cataldi, Ángel Adrián. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Labate, Carlos A.. Universidade de Sao Paulo; Brasil Fil: Rodrigues, Clenilson Martins. Ministerio da Agricultura Pecuaria e Abastecimento de Brasil. Empresa Brasileira de Pesquisa Agropecuaria; Brasil Fil: Talia, Paola Monica. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Se comparó la capacidad de ninfas de primer y tercer estadío de Delphacodes kuscheli Fennah (Hemiptera: Delphacidae) para adquirir y posteriormente transmitir el Mal de Río Cuarto virus (MRCV), bajo condiciones controladas. Ninfas I y III avirulíferas se alimentaron separadamente de plantas de trigo infectadas durante 48 horas, para luego ser colocadas en subgrupos de tres individuos sobre plantas de trigo sanas. Se realizaron transmisiones seriales utilizando períodos de inoculación de 24 horas. Ambos estadií os lograron adquirir y transmitir el MRCV, pero se evidenció una mayor cantidad de subgrupos infectivos cuando la adquisición se efectuó como ninfas I, así como una disminución significativa en la duración del período de latencia del virus respecto de los insectos que adquirieron el MRCV durante el tercer estadio ninfal.

O objetivo deste trabalho foi estabelecer uma tabela de vida para os estágios imaturos de Epinotia aporema, como parte de um estudo mais amplo para seu controle biológico. Os insetos foram criados em dieta artificial a 25±1ºC e 16:8 (luz:escuridão) horas de fotoperíodo. Para a identificação dos estágios larvais para estudos de interação inseto‑patógeno em condições de laboratório, as larguras de cápsula cefálica também foram determinadas. O período de incubação dos ovos foi de 4,13±0,30 dias, o estágio larval foi de 11,64±0,49 dias, e o tempo de desenvolvimento das pupas dependeu do sexo, com 8,51±0,69 dias para as fêmeas e 9,41±0,65 dias para os machos. Foram identificados cinco estágios larvais. The objective of this work was to establish a life table for the immature stages of Epinotia aporema, as part of a wider investigation on its biological control. Insects were reared on an artificial diet at 25±1ºC and a 16:8 (light:dark) hour photoperiod. For the identification of larval instars for the study of pathogen‑insect interactions under laboratory conditions, head capsule widths (HCWs) were also determined. The egg incubation period was 4.13±0.30 days, larval stage took 11.64±0.49 days, and the development time of the pupal phase was sex‑dependent with 8.51±0.69 days for the females and 9.41±0.65 days for the males. Five larval instars were identified. Fil: Arneodo Larochette, Joel Demián. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Quintana, Graciela. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Sciocco, Alicia Inés. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina

This is the first report of the association between the rose-grain aphid Metopolophium dirhodum, a potentially important cereal pest and the facultative symbiont Hamiltonella defensa. The infection with this gamma-proteobacterium was determined by PCR in laboratory-reared and field-collected specimens of an Argentinian population of the aphid. Partial bacterial 16S, IGS and 23S rRNA genes were sequenced and compared to other available Hamiltonella sequences by phylogenetic analysis. the present study provides new information on previously unknown M. dirhodum microbiota.

The Mal de Río Cuarto disease is caused by Mal de Río Cuarto virus (MRCV) transmitted by Delphacodes kuscheli. Comparative studies were carried out on the cytopathological alterations produced by MRCV in corn (Zea mays), wheat (Triticum aestivum) and barley (Hordeum vulgare), as seen with a transmission electron microscope. Corn plants were infected with viruliferous D. kuscheli collected from the endemic disease area (i.e. Río Cuarto County, Córdoba, Argentina). For the viral transmission to small grain cereal plants, laboratory rared insects were used. In this case, the inoculum source was wheat and barley plants infected with MRCV isolate grown in a greenhouse. Leaf samples with conspicuous symptoms were collected: enations and size reduction in corn; crenatures, swelling veins and dark green color in small grain cereals. Viral infection was corroborated by DAS-ELISA. Viroplasms containing complete and incomplete virus particles and fibrillar material were found in the cytoplasm of infected cells in all species. Mature virions were between 60 and 70 nm diameter. In wheat and barley, viroplasms and dispersed particles were observed only in phloem, while in corn virions were also found in cells of the bundle sheath. Crystalline arrays of particles were detected in corn enation constitutive cells. Tubular inclusions were found only in wheat samples. The three species showed abnormalities in the chloroplasts of affected cells. The results showed that MRCV cytopathology has similarities with other viruses from the genus Fijivirus, family family Reoviridae, but slight differences depending upon the host plant. La enfermedad denominada mal de Río Cuarto es causada por el Mal de Río Cuarto virus (MRCV) transmitido por Delphacodes kuscheli. Se reportan los resultados de un estudio de las alteraciones citopatológicas producidas por el MRCV en maíz (Zea mays), trigo (Triticum aestivum) y cebada (Hordeum vulgare), observadas en microscopio electrónico de transmisión. Las plantas de maíz fueron inoculadas con ejemplares infectivos de D. kuscheli colectados en la zona endémica de la enfermedad. Para la transmisión del virus a los cereales de grano fino se utilizaron insectos criados en laboratorio, siendo la fuente de inóculo plantas de trigo y cebada infectadas con un aislamiento del virus obtenido en Río Cuarto y mantenido en invernáculo. Se colectaron muestras de hojas con síntomas conspicuos en maíz, y en cereales de grano fino. La infección viral fue corroborada mediante DAS-ELISA. Se encontraron viroplasmas, conteniendo partículas virales completas e incompletas y material fibrilar, en el citoplasma de las células infectadas en todas las especies. Los viriones maduros midieron entre 60 y 70 nm de diámetro. En trigo y cebada, los viroplasmas y partículas dispersas fueron observados sólo en el floema, mientras que en maíz también se encontraron viriones en las células de la vaina. En maíz, se detectaron partículas formando arreglos cristalinos en las células de las enaciones. Exclusivamente en las muestras de trigo, se encontraron inclusiones tubulares. Las tres especies mostraron anomalías en los cloroplastos de las células afectadas. Los resultados obtenidos ponen de manifiesto que la citopatología propia del MRCV presenta semejanzas con la de otros fijivirus y pequeñas diferencias dependiendo de la planta hospedante.

Entre las enfermedades que afectan al cultivo de maíz (Zea mays) en Argentina, la producida por el virus del mal de Río Cuarto (MRCV) es la más importante. El MRCV pertenece a la familia Reoviridae, género Fijivirus, y su propagación en la naturaleza es realizada por Delphacodes kuscheli (Hemiptera: Delphacidae). La modalidad de transmisión para los miembros de este género de virus es persistente propagativa. Se estableció la necesidad de ajustar un sistema de transmisión eficiente del virus para estudios de caracterización, partiendo de poblaciones libres de virus criadas en laboratorio, para lo cual se ensayaron distintos períodos de adquisición, latencia e inoculación, evaluándose además un rango de hospedantes diferenciales. Se lograron obtener insectos libres de virus en cantidad suficiente para llevar a cabo los trabajos, mediante su cría en fitotrones y cámaras aclimatadas. La transmisión experimental del MRCV se efectuó exitosamente, bajo idénticas condiciones, empleando períodos de adquisición, latencia e inoculación de dos, 10 y uno día respectivamente para los cereales de grano fino y de dos, 10 y dos días para el maíz. Se infectaron de este modo las siguientes especies: maíz, cebada (Hordeum vulgare), avena (Avena sativa), trigo (Triticum aestivum), centeno (Secale cereale), grama rhodes (Chloris gayana) y alpiste (Phalaris canariensis). La detección del virus en las plantas inoculadas se efectuó mediante pruebas serológicas, análisis de dsRNA en electroforesis en gel de poliacrilamida (obteniéndose las 10 bandas típicas de los fijivirus) y microscopía electrónica, detectándose las partículas isométricas de entre 60 y 70 nm de diámetro. The disease caused by the "Mal de Rio Cuarto virus" (MRCV) is one of the most important diseases among those affecting maize (Zea mays) crops in Argentina. The MRCV belongs to the Reoviridae family, within the Fijivirus genus. It is spread in nature by Delphacodes kuscheli (Hemiptera: Delphacidae). The manner of transmission of the members of this virus genus is persistent propagative. An efficient virus transmission system was arranged in order to perform the various characterisation studies. These studies were carried out with healthy vector populations raised in the laboratory. Different acquisition, latency and inoculation periods were tested and a range of differential hosts was also evaluated, by breeding in phytotrons and controlled atmosphere chambers. A sufficient number of virus-free insects could thus be obtained to perform work. Experimental transmission of MRCV was successfully accomplished under identical conditions with acquisition, latency and inoculation periods of two, 10 and one days for fine grain cereals and two, 10 and two days for maize respectively. The following species were infected in this manner: maize (Zea mays), barley (Hordeum vulgare), oats (Avena sativa), wheat (Triticum aestivum), rye (Secale cereale), grama rhodes (Chloris gayana) and birdseed (Phalaris canariensis). Virus detection in inoculated plants was performed by means of serological tests, electrophoresis of the dsRNA in polyacrylamide gel (which showed the 10 typical bands of the fijiviruses) and electron microscopy, which showed 60 - 70 nm isometric particles.

Argentina is one of the leading soybean producing and exporting countries in the world. During the 2004/2005 cropping season, soybean production reached a record �8.� million tons. Although soybean is grown mainly in the central Pampas, it is also cultivated in the northwest of the country, where it plays a significant role in the local economy. Among the biotic factors affecting soybean yields in this region, fungal diseases account for the major economic losses (Wrather et al., Plant Dis. 81:107. 1��7). Nevertheless, diseases of viral etiology have also been recorded. Six different virus species have been detected infecting soybean in Argentina: Soybean mosaic virus (SMV), Alfalfa mosaic virus (AMV), Tobacco streak virus (TSV), Cucumber mosaic virus (CMV), Peanut mottle virus (PMV) and a geminivirus which has not been characterized yet. A survey conducted in Salta Province (NW Argentina) during late summer 2005 revealed the presence of soybean plants with systemic leaf chlorosis, distortion and stunting, which were negative in serological or hybridization tests to the viruses mentioned above. This work reports on the identification of the disease’s causal agent. Leaf dip preparations were made from leaves of symptomatic and asymptomatic (presumably healthy) soybean plants. Samples were ground in phosphate buffer pH 7 + 0.01% (w/v) sodium sulphite, stained with 2% uranyl acetate and examined under a JEOL 1220 electron microscope. Straight filamentous particles of about 6�0-650 nm, which resembled those of carlavirus, were consistently associated with the diseased plants, while no particles were observed in samples from asymptomatic soybeans. On this basis, plants were tested for Cowpea mild mottle virus (CPMMV, genus Carlavirus) by DAS-ELISA. Leaf samples were ground in extraction buffer (PBS pH 6.8 + 0.05% Tween 20 + 2% polyvinyl pyrrolidone) at a 1:5 (w/v) dilution. Specific polyclonal antisera (DSMZ GmbH, Germany) were used. Positive (supplied by DSMZ) and negative (healthy soybean) controls were included on each microtitre plate. After incubation with p-nitrophenyl phosphate at room temperature for 1 h, A405nm values greater than 0.�00 were obtained for the infected samples, compared with less than 0.100 for the healthy controls. CPMMV, a virus serologically unrelated to any other member of the genus, was first reported infecting Vigna unguiculata (L.) Walp. in Ghana (Brunt & Kenten, Ann. Appl. Biol. 74:67. 1�7�). Natural hosts of CPMMV include several species in the family Fabaceae. Its occurrence on soybean has been reported in southeast Asia, India and west Africa (Suryawanshi et al., Indian J. Virol. 5:12�. 1�8�). More recently, a disease causing stem necrosis of soybean in Brazil has also been attributed to CPMMV or a related carlavirus (Almeida et al., Fitopatol. Bras. �0:1�1. 2005). In Argentina, CPMMV has already been found on common bean (Phaseolus vulgaris L.) in Salta Province (RodriguezPardina et al., Australas. Plant Pathol. ��:12�. 2004), where large populations of the whitefly vector Bemisia tabaci Genn. may be responsible for the transmission of the pathogen to other crops. The possible spread of CPMMV to different soybean production regions, as well as its economic impact, still has to be determined.