Parasitism of the “Fuller’s rose weevil” Naupactus cervinus by Microctonus sp. in Argentina

We report the occurrence of an unidentified species of the wasp Microctonus Wesmael (Hymenoptera: Braconidae) parasitizing adults of the Fuller’s rose weevil Naupactus cervinus (Boheman) (Coleoptera: Curculionidae), a widespread pest of economically important crops included in the South American tribe Naupactini. Cytochrome c oxidase subunit I-based phylogenetic analysis indicates that the parasitoid is closely related to Microctonus hyperodae Loan. Their first instar larvae show slight morphological differences with this species. Superparasitism by first instar larvae occurred at low frequency. Some teratocytes were observed. Microctonus sp. and its host were infected with different strains of the reproductive parasite Wolbachia pipientis Hertig (Rickettsiales: Rickettsiaceae), although the bacterial lineage harbored by the wasp coincides with that infecting most parthenogenetic Naupactini. This multipartite association (weevil bearing both Wolbachia and Microctonus, and Microctonus bearing Wolbachia) opens challenging perspectives for future research on biological interactions and biological control.     

Biological characteristics of the egg phase of citrus root weevils

The goal of this work was to study some characteristics of the egg phase of three species of citrus root weevils. The insects were collected from citrus plants in Itapetininga, SP, and brought to the Laboratório de Biologia de Insetos of ESALQ/USP, in Piracicaba, SP, where the species Naupactus cervinus (Boheman), Naupactus versatilis (Hustache) and Parapantomorus fluctuosus (Boheman) were kept. Duration and viability of the egg phase were evaluated, and the lower temperature threshold and thermal constant (K) were calculated for these species. The species of citrus root weevils showed different duration of egg phases. The egg phase ranged from 40.4 to 13.8 N. cervinus, from 38.7 to 20.0 days for N. versatilis, and from 35.0 to 13.8 days for P. fluctuosus, depending upon temperature. The temperature thresholds of this stage were 8.1, 8.3, and 9.9 masculineC at thermal constant was 385.7, 397.7 and 294.1 degree-days, for N. cervinus, N. versatilis and P. fluctuosus respectively. The duration of the egg phases of N. cervinus and N. versatilis were similar at the same temperatures and P. fluctuosus had a faster development than Naupactus spp. in all temperatures tested.     

Subgenera of Charidotella Weise with description of a new subgenus and species from Brazil (Coleoptera,Chrysomelidae, Cassidinae,Cassidini)

A new subgenus and species, Charidotella (Chapadacassis subgen. n.) paradoxa sp. n. is described and figured from the Chapada plateau in Mato Grosso, Brazil. Subgenera of Charidotella Weise, 1896 are listed, supplemented with basic data, diagnostic table, and a key is proposed. Based on a study of respective type material following new combinations are proposed: Charidotella (Philaspis) stulta (Boheman, 1855), comb. n., Charidotella (Xenocassis) amoenula (Boheman, 1855), comb. n., Charidotella (Xenocassis) cyclographa (Boheman, 1855), comb. n., Charidotella (Xenocassis) discoidalis (Boheman, 1855), comb. n., Charidotella (Xenocassis) incerta (Boheman, 1855), comb. n., Charidotella (Xenocassis) purpurea (Linnaeus, 1758), comb. n., Charidotella (Xenocassis) myops (Boheman, 1855), comb. n. (all previously placed in the nominotypical subgenus), and Plagiometriona cingulata (Boheman, 1862), comb. n. (from Charidotella (Xenocassis)).     

Sudden death syndrome of soybean in South America is caused by four species of Fusarium: Fusarium brasiliense sp. nov., F. cuneirostrum sp. nov., F. tucumaniae, and F. virguliforme

Soybean sudden death syndrome (SDS) pathogens and dry bean root-rot pathogens were studied taxonomically, phylogenetically, and pathologically. Detailed phenotypic comparisons of macro- and microscopic features and phylogenetic analyses of multilocus DNA sequence data, including those on the nuclear ribosomal intergenic spacer region and the single copy nuclear gene translation elongation factor 1-a, indicated that they comprised five distinct species of Fusarium. Two new species causing soybean SDS in Brazil, F. brasiliense and F. cuneirostrum, are formally described. Fusarium cuneirostrum is responsible for soybean SDS in Brazil and dry bean or mung bean root-rot in the United States, Canada, and Japan. Strains of each species, including F. cuneirostrum isolates from dry bean and mung bean and F. phaseoli isolates from dry bean, were inoculated on soybean cultivar Pioneer 9492RR to determine their pathogenicity. Although intraspecific variation in pathogenicity was observed, all the species were able to induce typical SDS symptoms on soybean plants in the artificial inoculation tests. Comparisons of the key diagnostic morphological features reveal that all five species can be diagnosed using conidial morphology.     

RAPD and mitochondrial DNA analysis of the soybean stalk weevil, Sternechus subsignatus (Coleoptera: Curculionidae)

Sternechus subsignatus Boheman (Curculionidae: Sternechini) is one of the primary Curculionidae species that reduces soybean yield in Brazil. Initially, outbreaks were reported in southern Brazil in 1973; but, more recent, outbreaks were reported in Bahia (summer 1997-1998) and Maranh?o (summer 2003-2004), two states in northeastern Brazil. A putative related species, S. pinguis (Fabricius), was first detected in Salta Province, Argentina. The objective of this study was to evaluate intraspecific molecular polymorphisms of geographically distinct Sternechus populations. Randomly amplified polymorphic DNA (RAPD) profiles and partial mitochondrial cytochrome B (CytB) gene sequences were used to determine whether individual soybean stalk weevils were one of two different species and to infer pest invasion pattern. Putative S. pinguis and S. subsignatus populations were collected in San Agustin (Cruz Alta, Tucumán Province, Argentina) and different sampling sites in the Brazilian states of Paraná, Bahia and Maranh?o. Polymorphic bands were obtained by RAPD and analyzed by Dice coefficients. Populations from southern Brazil were more closely related genetically to an Argentinean group than the populations sampled in northeastern Brazil. The Londrina Co., Brazil population displayed the highest intra-population genetic similarity. Most of the soybean stalk weevils collected from San Agustin, Tucumán, Argentina were divergent from those collected in Brazil. Sequencing and parsimony analysis of CytB did not differentiate specimens collected in Argentina and Brazil. Thus, our data show that soybean stalk weevil outbreaks and population increases in northeastern Brazil involved local genotypes.     

Plant-Knowledge Adaptation in an Urban Setting: <Emphasis Type="Italic">Candomblé</Emphasis> Ethnobotany in New York City

Candomblé is an African-Brazilian religion that resulted from the adaptation of West African (especially Yoruba) beliefs in Brazil during and after the slave trade. This study seeks to understand the current evolution of Candomblé ethnobotanical knowledge as it travels from Brazil to New York City (NYC), therefore going through a second adaptation process. We identified which Brazilian plant species are still in use, which are being incorporated and/or replaced, and what factors are contributing to the ethnobotanical adaptation that is taking place in NYC. To accomplish this, we compiled an inventory of liturgical plants used by five highly skilled Candomblé practitioners living in NYC and then compared the vernacular and binomial Latin names of these plants to inventories previously published in Brazil by other authors. By doing this, we were able to distinguish patterns of knowledge continuity, assimilation, or substitution. Nearly two-thirds of the species identified in NYC’s inventory were cases of knowledge continuity, where most plants were used by at least four practitioners. Many of these frequently used species have survived the adaptation process from Africa to Brazil, and now from Brazil to NYC. Practitioners also assimilated (20%) and substituted (16%) some species. The assimilation process was mainly influenced by Santería, another Yoruba-derived religion widely practiced in NYC. Substitutions, however, were driven by two distinct forces. In one cohort (7%), species were morphologically and organoleptically similar to the original material, and replacements were mostly influenced by the easy accessibility of botanical materials. The other cohort (9%) was marked by a logical substitution process based on Yoruba rules of plant classification. Our results show that Candomblé practitioners in NYC are maintaining a notable level of cultural continuity, while cautiously assimilating new species and consciously or subconsciously replacing others. Although both accessibility of plant material and cultural forces play a role in the adaptation mechanism, the latter appears to be the most relevant to these highly skilled practitioners.     

Molecular Variability and Genetic Structure of Chrysodeixis includens (Lepidoptera: Noctuidae), an Important Soybean Defoliator in Brazil

This study provides the first genetic characterization of the soybean looper, Chrysodeixis includens (Walker, 1857), an important defoliating pest species of soybean crops in Brazil. Population genetic variability and the genetic structure of C. includens populations were evaluated by using ISSR markers with samples from the major soybean producing regions in Brazil in the growing seasons 2011/2012. Seven different primers were applied for population characterization of the molecular variability and genetic structure of 8 soybean looper populations from 8 states of Brazil. The seven ISSR loci generated 247 bands in 246 individuals of C. includens sampled. The expected heterozygosity (H _E) in the populations varied between 0.093 and 0.106, while the overall H _E was 0.099, indicating low genetic diversity. The analysis of molecular variance indicated that 98% of the variability was expressed among individuals within populations (F _ST = 0.021, p = 0.001). The low level of polymorphism over all populations, the high levels of gene flow, and the low genetic structure are indicatives of the exchange of genetic information between the different sampled regions. Population structuring suggests the presence of two major groups which do not correlate with their geographic sampling location in Brazil. These results may indicate recent recolonization of C. includens in Brazil or migration patterns following source-sink dynamics. Furthermore, the presence of two groups within C. includens suggests that a study on development of resistance or any other genetic-based trait needs to be evaluated on both groups, and pest management in soybean fields should be aware that differences may come to the control strategies they use.     

Temporal Variation in Seed Predation by Insects in a Population of Syagrus romanzoffiana (Arecaceae) in Santa Catarina Island, SC, Brazil

Insect seed predation may vary depending on seed production. The present study considers the hypothesis that the rates of seed predation tend to be smaller in years of higher fruit production. Thus, we monitored the production of fruits and predation of seeds of the palm Syagrus romanzoffiana over 2?years in the Atlantic Forest (Parque Municipal da Lagoa do Peri, Florianópolis, SC, Brazil), between July 2006 and June 2008. Plots of 0.25?m² were fitted under 20 mother plants and fruits were monthly collected for assessment of abundance and seed predation. There was variation in fruit production between the 2?years and among reproductive plants. Predation rates were high and occurred in the predispersal phase by the Curculionidae Revena rubiginosa Boheman, Anchylorhynchus aegrotus Fahraeus, and Anchylorhynchus variabilis Gyllenhal. Seed predation by these species of Anchylorhynchus is first registered in the present study. In average, about 60% of the seeds monthly produced in the population tend to escape insect predation in year of high or low production, becoming available for recruitment. The predation rate was not related to the amount of fruits produced per reproductive plant. Also, different than expected, there was a positive relation between the rates of seed predation and the total of fruits produced monthly on the plots. Thus, no evidence for the satiation of insect seed predators was found in this study with S. romanzoffiana.     

Analyses of the Xylem Sap Proteomes Identified Candidate Fusarium virguliforme Proteinacious Toxins

Background

Sudden death syndrome (SDS) caused by the ascomycete fungus, Fusarium virguliforme, exhibits root necrosis and leaf scorch or foliar SDS. The pathogen has never been identified from the above ground diseased foliar tissues. Foliar SDS is believed to be caused by host selective toxins, including FvTox1, secreted by the fungus. This study investigated if the xylem sap of F. virguliforme-infected soybean plants contains secreted F. virguliforme-proteins, some of which could cause foliar SDS development.

Results

Xylem sap samples were collected from five biological replications of F. virguliforme-infected and uninfected soybean plants under controlled conditions. We identified five F. virguliforme proteins from the xylem sap of the F. virguliforme-infected soybean plants by conducting LC-ESI-MS/MS analysis. These five proteins were also present in the excreted proteome of the pathogen in culture filtrates. One of these proteins showed high sequence identity to cerato-platanin, a phytotoxin produced by Ceratocystis fimbriata f. sp. platani to cause canker stain disease in the plane tree. Of over 500 soybean proteins identified in this study, 112 were present in at least 80% of the sap samples collected from F. virguliforme-infected and -uninfected control plants. We have identified four soybean defense proteins from the xylem sap of F. virguliforme-infected soybean plants. The data have been deposited to the ProteomeXchange with identifier PXD000873.

Conclusion

This study confirms that a few F. virguliforme proteins travel through the xylem, some of which could be involved in foliar SDS development. We have identified five candidate proteinaceous toxins, one of which showed high similarity to a previously characterized phytotoxin. We have also shown the presence of four soybean defense proteins in the xylem sap of F. virguliforme-infected soybean plants. This study laid the foundation for studying the molecular basis of foliar SDS development in soybean and possible defense mechanisms that may be involved in conferring immunity against F. virguliforme and other soybean pathogens.     

Symbiotic properties and first analyses of the genomic sequence of the fast growing model strain Sinorhizobium fredii HH103 nodulating soybean

Glycine max (soybean) plants can be nodulated by fast-growing rhizobial strains of the genus Sinorhizobium as well as by slow-growing strains clustered in the genus Bradyrhizobium. Fast-growing rhizobia strains with different soybean cultivar specificities have been isolated from Chinese soils and from other geographical regions. Most of these strains have been clustered into the species Sinorhizobium fredii. The S. fredii strain HH103 was isolated from soils of Hubei province, Central China and was first described in 1985. This strain is capable to nodulate American and Asiatic soybean cultivars and many other different legumes and is so far the best studied fast-growing soybean-nodulating strain. Additionally to the chromosome S. fredii HH103 carries five indigenous plasmids. The largest plasmid (pSfrHH103e) harbours genes for the production of diverse surface polysaccharides, such as exopolysaccharides (EPS), lipopolysaccharides (LPS), and capsular polysaccharides (KPS). The second largest plasmid (pSfrHH103d) is a typical symbiotic plasmid (pSym), carrying nodulation and nitrogen fixation genes. The present mini review focuses on symbiotic properties of S. fredii HH103, in particular on nodulation and surface polysaccharides aspects. The model strain S. fredii HH103 was chosen for genomic sequencing, which is currently in progress. First analyses of the draft genome sequence revealed an extensive synteny between the chromosomes of S. fredii HH103 and Rhizobium sp. NGR234.     

Advantageous attributes of larval whitefringed weevil, Naupactus leucoloma (Coleoptera: Curculionidae) for bioassaying soil fumigants, and responses to pure and plant-derived isothiocyanates

First instars of the soil-inhabiting whitefringed weevil, Naupactus leucoloma (Boheman), are a particularly good bioassay model for assessing volatile soil fumigants and biofumigants. Eggs are readily obtained and can be stored for long periods with larvae hatched on demand and the first instar is non-feeding, surviving without food or shelter. Longevity varies with temperature, but readily accommodates the period required to conduct bioassays without appreciable mortality of untreated controls. In vitro bioassays of pure methyl isothiocyanate, the active ingredient from metham sodium soil fumigant, and the less volatile 2-phenylethyl isothiocyanate, sensitively detected differences in toxicity and effects of temperature. Bioassay of volatiles emitted from hydrolysed tissue of various isothiocyanate-producing Brassica plants revealed widely varying toxicity effects, indicating that bioassays with N. leucoloma are a sensitive and relevant indicator of the potential of different plants for biofumigation of soil-borne pest organisms.     

Expression of soybean proteinase inhibitors in transgenic sugarcane plants: effects on natural defense against Diatraea saccharalis

The introduction and expression of proteinase inhibitor encoding genes into sugarcane (Saccharum officinarum L.) genome is an interesting strategy for conferring partial resistance to the sugarcane borer Diatraea saccharalis (Lepidoptera: Crambidae), the major insect pest of sugarcane in Brazil. To investigate the role of soybean (Glycine max L.) Kunitz trypsin inhibitor (SKTI) and soybean Bowman–Birk inhibitor (SBBI) in the control of D. saccharalis, the cDNAs encoding these proteinase inhibitors were placed under the control of the maize ubiquitin promoter (Ubi-1), and introduced into sugarcane callus using particle bombardment. Putative transgenic plants were initially identified after regeneration from callus growing in the presence of 30 mg l^–1 geneticin, while molecular characterization of transgenic plants revealed that both genes were incorporated into the sugarcane genome and expressed. We also carried out insect feeding trials using D. saccharalis neonate larvae and leaf tissue excised from propagated transgenic and untransformed plants, and found that the growth of larvae feeding on leaf tissue from transgenic plants containing BBI and Kunitz inhibitors was significantly retarded as compared to larvae fed on leaf tissue from untransformed plants. In greenhouse trials with transgenic sugarcane plants infested with D. saccharalis neonates, we found that these plants still presented the ‘dead heart’ symptom typically observed in susceptible plants in the field, suggesting that the retardation of the growth of D. saccharalis observed in the laboratory-based feeding trials was not sufficient to prevent this type of damage.     

Phenotypic, molecular phylogenetic, and pathogenetic characterization of Fusarium crassistipitatum sp. nov., a novel soybean sudden death syndrome pathogen from Argentina and Brazil

A novel soybean sudden death syndrome (SDS) pathogen from Argentina and Brazil is formally described herein as Fusarium crassistipitatum based on detailed phenotypic analyses of macro- and microscopic characters and phylogenetic analyses of multilocus DNA sequence data. Fusarium crassistipitatum can be distinguished from the other soybean SDS and bean (Phaseolus/Vigna) root rot pathogens (BRR) phenotypically by the production of yellowish colonies on PDA; and tall, stout, and mostly unbranched conidiophores with a thick-walled base, which form multiseptate conidia apically. Phylogenetic species recognition based on genealogical concordance of a six-gene dataset strongly supported the reciprocal monophyly of F. crassistipitatum with respect to the other SDS and BRR pathogens. Isolates of F. crassistipitatum were able to induce typical SDS foliar and root rot symptoms on soybean that were indistinguishable from those caused by three other SDS pathogens (i.e., F. virguliforme, F. brasiliense, and F. tucumaniae) on susceptible cultivars A-6445RG and N-4613RG in a pathogenicity experiment.     

Phylogeny of the tribe Naupactini (Coleoptera: Curculionidae) based on morphological characters

Naupactini (Curculionidae: Entiminae) is a primarily Neotropical tribe of broad‐nosed weevils with its highest genus and species diversity in South America. Despite several taxonomic contributions published during the last decades, the evolutionary history of Naupactini remains poorly understood. We present the first comprehensive phylogenetic analysis for this tribe based on a data matrix of 100 adult morphological characters scored for 70 species, representing 55 genera of Naupactini (ingroup) and four outgroups belonging to the entimine tribes Otiorhynchini, Entimini, Eustylini and Tanymecini. According to the most parsimonious tree Artipus does not belong to Naupactini; the genera with flat and broad antennae, formerly assigned to other entimine tribes, form a monophyletic group (Saurops (Curiades (Aptolemus (Platyomus)))) related to the clade (Megalostylus (Megalostylodes (Chamaelops Wagneriella))); and the genera distributed along the high Andes, Paramos and Puna form a natural group (Asymmathetes (Amphideritus (Leschenius (Amitrus (Obrieniolus (Melanocyphus Trichocyphus)))))), nested within a larger clade that includes Pantomorus, Naupactus and allied genera. Atrichonotus, Hoplopactus, Mimographus and Naupactus are not recovered as monophyletic. In order to address the taxonomic implications of our phylogenetic analysis, we propose the following nomenclatural changes: to transfer Artipus from Naupactini to Geonemini, to revalidate the genera Mimographopsis (type species M. viridicans), and to revalidate the genus Floresianus (type species F. sordidus). The evolution of selected characters is discussed. This published work has been registered in ZooBank, http://zoobank.org/urn:lsid:zoobank.org:pub:C8AA4388‐A2F0‐4E2D‐889A‐500BEA5A9DE1 .     

High Susceptibility to Cry1Ac and Low Resistance Allele Frequency Reduce the Risk of Resistance of Helicoverpa armigera to Bt Soybean in Brazil

The Old World bollworm, Helicoverpa armigera (Hübner), was recently introduced into Brazil, where it has caused extensive damage to cotton and soybean crops. MON 87701 × MON 89788 soybean, which expresses the Bt protein Cry1Ac, was recently deployed in Brazil, providing high levels of control against H. armigera. To assess the risk of resistance to the Cry1Ac protein expressed by MON 87701 × MON 89788 soybean in Brazil, we conducted studies to evaluate the baseline susceptibility of H. armigera to Cry1Ac, in planta efficacy including the assessment of the high-dose criterion, and the initial resistance allele frequency based on an F₂ screen. The mean Cry1Ac lethal concentration (LC₅₀) ranged from 0.11 to 1.82 μg·mL⁻¹ of diet among all H. armigera field populations collected from crop seasons 2013/14 to 2014/15, which indicated about 16.5-fold variation. MON 87701 × MON 89788 soybean exhibited a high level of efficacy against H. armigera and most likely met the high dose criterion against this target species in leaf tissue dilution bioassays up to 50 times. A total of 212 F₂ family lines of H. armigera were established from field collections sampled from seven locations across Brazil and were screened for the presence of MON 87701 × MON 89788 soybean resistance alleles. None of the 212 families survived on MON 87701 × MON 89788 soybean leaf tissue (estimated allele frequency = 0.0011). The responses of H. armigera to Cry1Ac protein, high susceptibility to MON 87701 × MON 89788 soybean, and low frequency of resistance alleles across the main soybean-producing regions support the assumptions of a high-dose/refuge strategy. However, maintenance of reasonable compliance with the refuge recommendation will be essential to delay the evolution of resistance in H. armigera to MON 87701 × MON 89788 soybean in Brazil.     

A new species of Philometra Costa, 1845 (Nematoda: Philometridae) from the freshwater fish (red piranha) Pygocentrus nattereri Kner (Characidae) in Amazonia, Brazil

A new nematode species, Philometra nattereri n. sp. (Philometridae), is described from female specimens found in the oculo-orbits and nasal cavity of the red piranha Pygocentrus nattereri Kner (Characiformes: Characidae) from five lakes in Central Amazonia, Brazil, collected in 2008 and 2009 (overall prevalence 12%, intensity 1?C3 nematodes per fish). Based on light and scanning electron microscopical examination, the new species differs from most other congeners parasitising freshwater fishes in that its oesophageal gland extends anteriorly far anterior to the level of the nerve-ring, in the presence of 14 small cephalic papillae arranged in two circles and in having two minute caudal projections. This is the first species of Philometra Costa, 1845 reported from fishes of the family Characidae and the second valid species of this genus parasitic in freshwater fishes of Brazil and South America.     

Spatial Distribution of Adult <Emphasis Type="Italic">Anthonomus grandis</Emphasis> Boheman (Coleoptera: Curculionidae) and Damage to Cotton Flower Buds Due to Feeding and Oviposition

The cotton boll weevil Anthonomus grandis Boheman (Coleoptera: Curculionidae) is the main pest in cotton crop around the world, directly affecting cotton production. In order to establish a sequential sampling plan, it is crucial to understand the spatial distribution of the pest population and the damage it causes to the crop through the different developmental stages of cotton plants. Therefore, this study aimed to investigate the spatial distribution of adults in the cultivation area and their oviposition and feeding behavior throughout the development of the cotton plants. The experiment was conducted in Maracaju, Mato Grosso do Sul, Brazil, in the 2012/2013 and 2013/2014 growing seasons, in an area of 10,000 m², planted with the cotton cultivar FM 993. The experimental area was divided into 100 plots of 100 m² (10 × 10 m) each, and five plants per plot were sampled weekly throughout the crop cycle. The number of flower buds with feeding and oviposition punctures and of adult A. grandis was recorded throughout the crop cycle in five plants per plot. After determining the aggregation indices (variance/mean ratio, Morisita’s index, exponent k of the negative binomial distribution, and Green’s coefficient) and adjusting the frequencies observed in the field to the distribution of frequencies (Poisson, negative binomial, and positive binomial) using the chi-squared test, it was observed that flower buds with punctures derived from feeding, oviposition, and feeding + oviposition showed an aggregated distribution in the cultivation area until 85 days after emergence and a random distribution after this stage. The adults of A. grandis presented a random distribution in the cultivation area.