Phylogeography Approach of Diloboderus abderus (Coleoptera: Melolonthidae) in the Southern Cone of America.

Diloboderus abderus (Sturm, 1826) (Coleoptera: Melolonthidae) is a serious soil pest of corn, wheat, oat, and natural and cultivated pastures in Argentina, Paraguay, Uruguay, and southern Brazil. Despite its economic importance, the genetic diversity and population structure of D. abderus remain unknown. We sequenced a fragment of the mitochondrial gene cytochrome oxidase I region (COI), of six populations of D. abderus from the Southern Cone of America. The mtDNA marker revealed a high haplotype diversity, high pairwise FST values, and significant genetic variations among populations. No correlation was found between genetic and geographical distances, yet the most common haplotype (Dab01) was present in four out of the six populations. Analysis of molecular variance showed that most of the variation was within populations of D. abderus. Tajima's D and Fu's FS tests indicated no evidence that D. abderus populations are under recent expansion. Our results indicate that genetic-based traits will likely remain localized or spread slowly, and management strategies need to be undertaken on a small scale.

First record of soybean stem fly Melanagromyza sojae (Diptera: Agromyzidae) in Paraguay confirmed by molecular evidence.

We provided the first scientific record of Melanagromyza sojae (Zehntner, 1900), through molecular characterization of partial mtDNA COI gene, that confirms the occurrence of this pest in Paraguay. Previously reported in Brazil, an outbreak of larvae of M. sojae known as the soybean stem fly (SSF) that belongs to the family Agromyzidae, was also noted in soybean fields from the Canindeyú, Alto Paraná and Itapúa Departments in Paraguay. This pest is highly polyphagous, attacking various host plant species from the family Fabaceae, such as soybean and other beans. The implications of SSF detection in Paraguay are discussed in relation to the current soybean cultivation practices from this agriculturally important South American region, including Brazil.

Soybean Stem Fly, Melanagromyza sojae (Diptera: Agromyzidae), in the New World: detection of high genetic diversity from soybean fields in Brazil.

Soybean Stem Fly (SSF), Melanagromyza sojae (Zehntner), belongs to the family Agromyzidae and is highly polyphagous, attacking many plant species of the family Fabaceae, including soybean and other beans. SSF is regarded as one of the most important pests in soybean fields of Asia (e.g., China, India), North East Africa (e.g., Egypt), parts of Russia, and South East Asia. Despite reports of Agromyzidae flies infesting soybean fields in Rio Grande do Sul State (Brazil) in 1983 and 2009 and periodic interceptions of SSF since the 1940s by the USA quarantine authorities, SSF has not been officially reported to have successfully established in the North and South Americas. In South America, M. sojae was recently confirmed using morphology and its complete mitochondrial DNA (mtDNA) was characterized. In the present study, we surveyed the genetic diversity of M. sojae, collected directly from soybean host plants, using partial mtDNA cytochrome oxidase I (COI) gene, and provide evidence of multiple (>10) maternal lineages in SSF populations in South America, potentially representing multiple incursion events. However, a single incursion involving multiple-female founders could not be ruled out. We identified a haplotype that was common in the fields of two Brazilian states and the individuals collected from Australia in 2013. The implications of SSF incursions in southern Brazil are discussed in relation to the current soybean agricultural practices, highlighting an urgent need for better understanding of SSF population movements in the New World, which is necessary for developing effective management options for this significant soybean pest.

Mitochondrial DNA COI characterization of Helicoverpa armigera (Lepidoptera: Noctuidae) from Paraguay and Uruguay.

Since its detection in Brazil in 2013, the Old World cotton bollworm Helicoverpa armigera has been reported in Argentina, Paraguay, and Bolivia. Here we present evidence extending the South American range of H. armigera to Uruguay, using polymerase chain reaction and sequencing of the partial mitochondrial DNA (mtDNA) cytochrome oxidase I region. Molecular characterization of this gene region from individuals from Paraguay also supports previous morphological identification of H. armigera in Paraguay. Shared mtDNA haplotypes in H. armigera from Brazil, Uruguay, and Paraguay were identified. Additional surveying of populations in this region will be imperative to better monitor and understand factors that are underpinning its presence and successful adaptation in these South American regions. We discuss our findings with respect to the development of resistance pest management strategies of this invasive insect pest in a predominantly monoculture soybean crop landscape in the Southern Cone region.

Comparative analysis of protocols for DNA extraction from soybean caterpillars.

Genomic DNA extraction is crucial for molecular research, including diagnostic and genome characterization of different organisms. The aim of this study was to comparatively analyze protocols of DNA extraction based on cell lysis by sarcosyl, cetyltrimethylammonium bromide, and sodium dodecyl sulfate, and to determine the most efficient method applicable to soybean caterpillars. DNA was extracted from specimens of Chrysodeixis includens and Spodoptera eridania using the aforementioned three methods. DNA quantification was performed using spectrophotometry and high molecular weight DNA ladders. The purity of the extracted DNA was determined by calculating the A260/A280 ratio. Cost and time for each DNA extraction method were estimated and analyzed statistically. The amount of DNA extracted by these three methods was sufficient for PCR amplification. The sarcosyl method yielded DNA of higher purity, because it generated a clearer pellet without viscosity, and yielded high quality amplification products of the COI gene I. The sarcosyl method showed lower cost per extraction and did not differ from the other methods with respect to preparation times. Cell lysis by sarcosyl represents the best method for DNA extraction in terms of yield, quality, and cost effectiveness.

Ecological Characterization of White Grubs (Coleoptera: Melolonthidae) Community in Cultivated and Noncultivated Fields.

Comparative studies on the density and diversity of white grubs community (Coleoptera: Melolonthidae) occurring in cultivated and noncultivated fields of the Planalto region of the state of Rio Grande do Sul, Brazil, are presented. Sampling was carried out in 23 municipalities during the 2009 and 2010 winter seasons. Cultivated and noncultivated fields were chosen in each locality. Melolontid larvae were collected for identification and counted to determine the population density. A mean of 12.9 larvae m(-2) were collected in cultivated areas against 10.5 larvae m(-2) in noncultivated areas. The latter were more diverse (H' = 2.52) than cultivated areas (H' = 2.26). Despite the high evenness index (J = 0.75 noncultivated and J = 0.74 cultivated), faunistic parameters indicated Cyclocephala flavipennis Arrow and Diloboderus abderus Sturm as an extremely dominant species in cultivated areas. These results showed that the population density of white grubs increases, and their community composition is affected in cultivated areas.

On the presence of five species of naupactini (Coleoptera: Curculionidae) damaging soybean in Brazil.

We report five broad-nosed weevils of the tribe Naupactini, Naupactus leucoloma Boheman, Naupactus minor (Buchanan), Naupactus peregrinus (Buchanan), Naupactus tremolerasi Hustache and Pantomorus viridisquamosus (Boheman) (Coleoptera: Curculionidae) damaging soybean plants in the state of Rio Grande do Sul, Brazil. Some of these species are recorded for the first time in Brazil, and they all belong to species groups naturally distributed in the prairies of southern Brazil, Uruguay, Central Argentina and Paraguay. Three of them have been introduced, established and expanded in other countries outside South America. The five species in conjunction with other native species of Naupactini could cause severe damages to soybean crops if the root-feeding larvae attack young growing plants.

First record of Teratopactus nodicollis (Coleoptera: Curculionidae) in dry bean (Phaseolus vulgaris).

Observations on the bioecology and damage of Teratopactus nodicollis Boheman on Phaseolus vulgaris were carried out on field samples by assessing the number of larvae and root damage in 40 ha of a dry bean field from the Federal District, Brazil (16°4'28.41"W; 47°30'21.13"S). Larvae caused the greatest damage at the stage of germination, emergence, and primary leaves, producing 50 % stand reduction. Most larvae pupated in August and September, and adult emergence occurred in middle October. Some larvae were infected with the fungus Metarhizium spp., a biological agent that would be naturally controlling this insect.

Wolbachia infection in the tribe Naupactini (Coleoptera, Curculionidae): association between thelytokous parthenogenesis and infection status.

Several parthenogenetic species of broad-nosed weevils exist, some of them of economic importance because of their pest status. Screening of the maternally inherited Wolbachia bacterium in 29 weevils of the tribe Naupactini, using multilocus sequence typing allowed us to assess a significant correlation between asexuality and infection, and suggests an involvement of Wolbachia in the origin of this reproductive mode. The nine Wolbachia strains retrieved from the Naupactini belong to the B supergroup. Phylogenetic analysis of these strains, along with other 23 strains obtained from arthropods and nematodes, supports previous hypotheses that horizontal transfer of Wolbachia amongst species from unrelated taxa has been pervasive.

Platynota rostrana (Walker) (Tortricidae) and Phidotricha erigens Raganot (Pyralidae): artificial diet effects on biological cycle

The lepidopterans Platynota rostrana (Walker) (Tortricidae) and Phidotricha erigens Raganot (Pyralidae) have been found frequently in citrus groves in São Paulo State in recent years. Since in Brazil, the fertility cycle of these two species is largely unknown, as are details of the damage wrought by them in crops, this research studied these aspects of the two species, which were kept under laboratory conditions (temperature 25 ± 2 °C, 70 ± 10 percent RH, 14 h photophase) and on an artificial diet. The duration of the biological cycle (egg-adult) for P. rostrana was 38.3 days and total viability was 44.0 percent; for P. erigens these values were 32.5 days and 63.6 percent, respectively. Both species showed five larval instars. Females of P. rostrana laid an average of 308 eggs, whereas those of P. erigens laid an average of 106 eggs. In both species, female pupae were heavier than males. Male and female longevity for both species was nearly 10 days. Based on the data obtained, the artificial diet produced better results in P. rostrana than in P. erigens. If these species, which have the potential to reach pest status in the citrus groves of São Paulo State, could be reared on an artificial diet, research on their control by alternative methods would be easier.

Os lepidópteros Platynota rostrana (Walker) (Tortricidae) e Phidotricha erigens Raganot (Pyralidae) têm sido constatados com freqüência nos pomares cítricos do Estado de São Paulo, nos últimos anos. O objetivo deste trabalho foi estudar a biologia das duas espécies, em condições de laboratório (temperatura 25 ± 2 °C, UR. 70 ± 10 por cento e fotofase de 14 h) em dieta artificial, elaborar uma tabela de vida de fertilidade e descrever os danos causados no campo, devido ao desconhecimento destes aspectos biológicos dos referidos insetos no Brasil. A duração do ciclo biológico (ovo-adulto) de P. rostrana foi de 38,3 dias e a viabilidade total de 44,0 por cento, enquanto para P. erigens foi de 32,5 dias e 63,6 por cento, respectivamente. Ambas as espécies apresentaram cinco ínstares e as pupas de fêmeas foram mais pesadas do que as de machos. As fêmeas de P. rostrana colocaram, em média, 308 ovos e as de P. erigens 106 ovos. A longevidade de machos e fêmeas das duas espécies foi próxima de 10 dias. Pela tabela de vida de fertilidade concluiu-se que P. rostrana tem melhor desempenho em dieta artificial que P. erigens. É possível criar estas espécies em dieta artificial, facilitando o desenvolvimento de pesquisas relacionadas ao seu controle por métodos alternativos, caso elas assumam o status de pragas nos pomares de São Paulo.

Platynota rostrana (Walker) (Tortricidae) AND Phidotricha erigens Raganot (Pyralidae): artificial diet effects on biological cycle.

The lepidopterans Platynota rostrana (Walker) (Tortricidae) and Phidotricha erigens Raganot (Pyralidae) have been found frequently in citrus groves in São Paulo State in recent years. Since in Brazil, the fertility cycle of these two species is largely unknown, as are details of the damage wrought by them in crops, this research studied these aspects of the two species, which were kept under laboratory conditions (temperature 25 +/- 2 degrees C, 70 +/- 10% RH, 14 h photophase) and on an artificial diet. The duration of the biological cycle (egg-adult) for P. rostrana was 38.3 days and total viability was 44.0%; for P. erigens these values were 32.5 days and 63.6%, respectively. Both species showed five larval instars. Females of P. rostrana laid an average of 308 eggs, whereas those of P. erigens laid an average of 106 eggs. In both species, female pupae were heavier than males. Male and female longevity for both species was nearly 10 days. Based on the data obtained, the artificial diet produced better results in P. rostrana than in P. erigens. If these species, which have the potential to reach pest status in the citrus groves of São Paulo State, could be reared on an artificial diet, research on their control by alternative methods would be easier.