Performance of Sirex noctilio’s biocontrol agent Deladenus siricidicola, in known and predicted hosts

Survival of the free-living mycetophagous form of Deladenus siricidicola, the major biological control agent of Sirex woodwasp, Sirex noctilio, was tested in known (Pinus taeda) and predicted novel (P. elliottii subsp. elliottii × P. caribaea var. hondurensis) hybrid host taxa. Trials were established in the field to simulate nematode dispersal both naturally by infected wasps and following commercial inoculation, as well as in the laboratory under controlled conditions. Nematodes showed reduced survival in hybrid pine compared with P. taeda for all tree-associated treatments, but performed equivalently in petri-dish bioassays containing substrate of each taxon. Growth of Amylostereum areolatum, the food source of D. siricidicola was lower on plates containing ground hybrid substrate than on plates containing ground P. taeda. Some physical differences were found between taxa, including differences in bordered pit diameters, tracheid widths, and basic density, but these did not consistently explain reduced performance. More plant secondary compounds (predominantly oleoresins) were present in hybrid taxa than in P. taeda, and in standing trees compared with felled trees. Our results suggested that D. siricidicola may not be as effective in hybrid pine taxa for the biological control of S. noctilio as it is in its current known host taxa, possibly because of reduced growth of its food source, A. areolatum in hybrid pine.

Seafloor heterogeneity influences the biodiversity–ecosystem functioning relationships in the deep sea

Theoretical ecology predicts that heterogeneous habitats allow more species to co-exist in a given area. In the deep sea, biodiversity is positively linked with ecosystem functioning, suggesting that deep-seabed heterogeneity could influence ecosystem functions and the relationships between biodiversity and ecosystem functioning (BEF). To shed light on the BEF relationships in a heterogeneous deep seabed, we investigated variations in meiofaunal biodiversity, biomass and ecosystem efficiency within and among different seabed morphologies (e.g., furrows, erosional troughs, sediment waves and other depositional structures, landslide scars and deposits) in a narrow geo-morphologically articulated sector of the Adriatic Sea. We show that distinct seafloor morphologies are characterized by highly diverse nematode assemblages, whereas areas sharing similar seabed morphologies host similar nematode assemblages. BEF relationships are consistently positive across the entire region, but different seabed morphologies are characterised by different slope coefficients of the relationship. Our results suggest that seafloor heterogeneity, allowing diversified assemblages across different habitats, increases diversity and influence ecosystem processes at the regional scale, and BEF relationships at smaller spatial scales. We conclude that high-resolution seabed mapping and a detailed analysis of the species distribution at the habitat scale are crucial for improving management of goods and services delivered by deep-sea ecosystems.

Étude de la toxicité causée par le gène C9orf72 dans la Sclérose Latérale Amyotrophique

La Sclérose Latérale Amyotrophique (SLA) est une maladie neurodégénérative qui affecte les neurones moteurs. 10% des cas sont des cas familiaux et l’étude de ces familles a mené à la découverte de plusieurs gènes pouvant causer la SLA, incluant SOD1, TARDBP et FUS. L’expansion de la répétition GGGGCC dans le gène C9orf72 est, à ce jour, la cause la plus connue de SLA. L’impact de cette expansion est encore méconnu et il reste à déterminer si la toxicité est causée par un gain de fonction, une perte de fonction ou les deux. Plusieurs gènes impliqués dans la SLA sont conservés entre le nématode Caenorhabditis elegans et l’humain. C. elegans est un vers transparent fréquemment utilisé pour des études anatomiques, comportementales et génétiques. Il possède une lignée cellulaire invariable qui inclue 302 neurones. Aussi, les mécanismes de réponse au stress ainsi que les mécanismes de vieillissement sont très bien conservés entre ce nématode et l’humain. Donc, notre groupe, et plusieurs autres, ont utilisé C. elegans pour étudier plusieurs aspects de la SLA. Pour mieux comprendre la toxicité causée par l’expansion GGGGCC de C9orf72, nous avons développé deux modèles de vers pour étudier l’impact d’une perte de fonction ainsi que d’un gain de toxicité de l’ARN. Pour voir les conséquences d’une perte de fonction, nous avons étudié l’orthologue de C9orf72 dans C. elegans, alfa-1 (ALS/FTD associated gene homolog). Les vers mutants alfa-1(ok3062) développent des problèmes moteurs causant une paralysie et une dégénérescence spécifique des neurones moteurs GABAergiques. De plus, les mutants sont sensibles au stress osmotique qui provoque une dégénérescence. D’autre part, l’expression de la séquence d’ARN contenant une répétition pathogénique GGGGCC cause aussi des problèmes moteurs et de la dégénérescence affectant les neurones moteurs. Nos résultats suggèrent donc qu’un gain de toxicité de l’ARN ainsi qu’une perte de fonction de C9orf72 sont donc toxiques pour les neurones. Puisque le mouvement du vers peut être rapidement évalué en cultivant les vers dans un milieu liquide, nous avons développé un criblage de molécules pouvant affecter le mouvement des vers mutants alfa-1 en culture liquide. Plus de 4 000 composés ont été évalués et 80 ameliore la mobilité des vers alfa-1. Onze molécules ont aussi été testées dans les vers exprimant l’expansion GGGGCC et huit diminuent aussi le phénotype moteur de ces vers. Finalement, des huit molécules qui diminent la toxicité causée par la perte de fonction de C9orf72 et la toxicité de l’ARN, deux restaurent aussi l’expression anormale de plusieurs transcrits d’ARN observée dans des cellules dérivées de patient C9orf72. Avec ce projet, nous voulons identifier des molécules pouvant affecter tous les modes de toxicité de C9orf72 et possiblement ouvrir de nouvelles avenues thérapeutiques

Prácticas culturales para el manejo de la Sigatoka negra en bananos y plátanos

Black sigatoka (Mycosphaerella fijiensis) control consists in the continuous use of fungicide sprays and cultural practices. Cultural control reduces inoculum sources of the pathogen and favorable environmental conditions for its development, as well as it increases the vigor of the plants. In order to manage the disease it is necessary to know its behavior through the time, its relationship with the weather and the crop management. The most important cultural practice to reduce the inoculum source is the removal of the whole affected leaves or only the diseased portions of the leaves. The removed tissue is placed over the ground and the use of urea is feasible to accelerate its decomposition. An alternative practice is the "mini-composting" that consists in forming small heaps with the dead leaves and harvested plants for its quick degradation, which allows to reduce the pathogen inoculum and to incorporate nutrients and organic matters into the soil. The early pruning of the tips of the Young leaves before they show symptoms of the disease and the quick elimination of harvested plants reduce the inoculum. Also, the agronomic management of the crop helps to reduce the favorable conditions and to increase the vigor of the plants: this management considers plantation density, drainage systems, irrigation methods, control of weeds, chemical-biological fertilization, and nematode control.

Aspectos sanitários do cultivo da alface americana, irrigada com águas recptoras de efluentes urbanos

Avaliou-se a influência da disposição de mangueiras gotejadoras nos canteiros e a injeção ou não de cloro na água de irrigação, nas condições sanitárias do solo e da alface americana irrigada (Lactuca sativa L.) com águas receptoras de efluentes urbanos. Foram realizadas análises microbiológicas de amostras de água do solo e da alface, no decorrer de todo o ciclo de cultivo. Objetivou-se determinar a possível existência de Salmonella spp. e de formas evolutivas de parasitos humanos e a quantidade de coliformes fecais, em pontos e épocas diferentes do experimento, impedindo assim o consumo da alface. Os resultados não indicaram a presença dos dois primeiros em nenhuma das amostras, mas sim de parasitos não humanos (nematóides) de vida livre no solo. em relação à quantidade de coliformes fecais (NMP ml-1), o valor encontrado na cultura atende às exigências da Secretaria de Vigilância Sanitária do Ministério da Saúde brasileiro, porém a presença dos nematóides em quantidades superiores ao permitido pela Organização Mundial de Saúde (OMS) inviabiliza o seu consumo.

First record of molluscs naturally infected with Angiostrongylus cantonensis (Chen, 1935) (Nematoda: Metastrongylidae) in Brazil

Seeking the identification of Angiostrongylus cantonensis as a potential etiological agent of three clinical cases of eosinophilic meningitis, mollusc specimens were collected in the state of Espírito Santo, Brazil. The snails were identified as Sarasinula marginata (45 specimens), Subulina octona (157), Achatina fulica (45) and Bradybaena similaris (23). Larvae obtained were submitted to polymerase chain reaction and restriction fragment length polymorphism diagnosis. Their genetic profile were corresponded to A. cantonensis. Rattus norvegicus experimentally infected with third-stage larvae, developed menigoencephalitis, and parasites became sexually mature in the lungs. Additionally, larvae obtained from A. fulica snails, from São Vicente, state of São Paulo, also showed genetic profiles of this nematode. This is the first record of Brazilian molluscs infected with this nematode species.

Melhoramento genético florestal e o desafio imposto pela doença da murchidão do pinheiro

Mestrado em Engenharia Florestal e dos Recursos Naturais - Instituto Superior de Agronomia - UL

Formulación bioinsecticida a partir del aceite esencial de ambrosia arborescens mill (altamisa) de aplicación canina

La presente investigación se planteó reemplazar el uso de insecticidas sintéticos, formulando un champú bioinsecticida de aplicación canina mediante la acción biocida del aceite esencial deAmbrosia arborescens Mill (Altamisa). La planta se recolectó en las laderas del rio Tomebamba, cercanas al Campus Balzay de la Universidad de Cuenca Parroquia San Joaquín. La recolección se realizó durante los meses de Enero a Marzo del 2016. El desarrollo y formulación del producto se realizó en el Laboratorio de Biotecnología, Facultad de Ciencias Químicas de la Universidad de Cuenca. La obtención del aceite esencial de A. arborescens se realizó mediante hidrodestilación por el método Clevenger, con un rendimiento del 0,14%. La actividad biocida se estableció en un ensayo “in vitro” ante el nematodo Panagrellus redivirus, determinándose la dosis letal (DL50) de 250 uL/mL. Debido a la moderada DL50y bajo rendimiento, se planteó como estrategia, determinar el DL50 del extracto orgánico de A. arborescens, el cual se obtuvo mediante una extracción con metanol, consiguiendo un rendimiento del 2 % y DL50de 31,25 uL/mL. De acuerdo estos resultados se procedió a realizar pruebas en pulgas de perros(Ctenocephalides canis) con el extracto de A. arborescens, estableciendo una efectividad del 100 % a la concentración de 46,875 mg/mL en el periodo de tiempo más corto, siendo esta la dosis aplicada para la formulación del champú. El extracto metanólico de A. arborescens presentó elevada actividad biocida, comparado con el aceite esencial. Esta sustancia activa es promisoria en la formulación de bioinsecticidas para mascotas.

Evaluation of the reaction of watermelon parent and F1 plants to Meloidogyne enterolobii.

The aim of this study was to evaluate the performance of progenies from Citrullus lanatus var. lanatus (cultivated watermelons) when crossed with progenies from C. lanatus var. citroides (fodder watermelon with a historic of resistance to the nematode Meloidogyne enterolobii). The parents and their F1s were evaluated for resistance to this nematode. In the initial stages of eleven treatments, watermelon seedlings plantlets were transplanted to plastic bags of six kilograms once the first leaves developed. Ten inoculated plants with 5,200 eggs in the soil near the stem of the plant and four non-inoculated ones were used in each treatment, in a complete block design. Sixty-two days after sowing, the following characteristics were evaluated: the length of the aerial part of the plant (LAP, in m), fresh mass of the aerial part (FMAP, in g), root fresh mass (RFM, in g), egg number (EN) and reproduction factor (RF). A comparison between the averages of inoculated and non-inoculated plants was performed using Scott-Knott test at 5% and the diallelic analysis was performed using the GENES program. The morphological characteristics did not allow for the identification of the parent plants or the F1s with respect to nematode resistance, but the variables EN and RF were useful for such identification. The analyses of the general and specific combining abilities indicate highly significant effects with respect to this resistance, showing additive gene effects as well as dominance and epistatic gene effects, allowing for identification of parents and F1s that can be used in watermelon breeding programs to improve resistance to the M. enterolobii.

Genome selection in fruit breeding: application to table grapes.

Genomic selection (GS) has recently been proposed as a new selection strategy which represents an innovative paradigm in crop improvement, now widely adopted in animal breeding. Genomic selection relies on phenotyping and high-density genotyping of a sufficiently large and representative sample of the target breeding population, so that the majority of loci that regulate a quantitative trait are in linkage disequilibrium with one or more molecular markers and can thus be captured by selection. In this study we address genomic selection in a practical fruit breeding context applying it to a breeding population of table grape obtained from a cross between the hybrid genotype D8909-15 (Vitis rupestris × Vitis arizonica/girdiana), which is resistant to dagger nematode and Pierce?s disease (PD), and ?B90-116?, a susceptible Vitis vinifera cultivar with desirable fruit characteristics. Our aim was to enhance the knowledge on the genomic variation of agronomical traits in table grape populations for future use in marker-assisted selection (MAS) and GS, by discovering a set of molecular markers associated with genomic regions involved in this variation. A number of Quantitative Trait Loci (QTL) were discovered but this method is inaccurate and the genetic architecture of the studied population was better captured by the BLasso method of genomic selection, which allowed for efficient inference about the genetic contribution of the various marker loci. The technology of genomic selection afforded greater efficiency than QTL analysis and can be very useful in speeding up the selection procedures for agronomic traits in table grapes.

Espécies de Meloidogyne em hortaliças e outras culturas provenientes de áreas periurbanas da África Subsaariana

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Departamento de Fitopatologia, Programa de Pós-Graduação em Fitopatologia, 2016.

Multiplicação do nematoide Meloidogyne paranaensis e velocidade de enraizamento de estacas caulinares em sete espécies de plantas medicinais

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Agronomia e Medicina Veterinária, Programa de Pós-Graduação em Agronomia, 2016.

Nematóide (Heterodera glycines Ichinohe) formador de cisto em soja.

O nematóide Heterodera glycines Ichinohe é considerado o parasita mais destrutivo da soja (Glycine max (L.) Merrill). A doença causada pela infestação desse nematóide, primeiramente obserada no Japão, recebeu a denominção de "nanismo amarelo", devido aos sintomas por ela produzidos nas plantas de soja infecatadas.

Development of a novel molecular tool for the rapid assessment of the biodiversity changes of benthic nematodes assemblages. Mares Conference Marine Ecosystems Helath and conservation

The molecular profiling system was developed using directed terminal-restriction fragment length polymorphism (dT-RFLP) to characterize soil nematode assemblages by relative abundance of feeding guilds and validation by comparison to traditional morphological method. The good performance of these molecular tools applied to soil nematodes assemblages create an opportunity to develop a novel approach for rapid assessment of the biodiversity changes of benthic nematodes assemblages of marine and estuarine sediments. The main aim of this research is to combine morphological and molecular analysis of estuarine nematodes assemblages, to establish a tool for fast assessment of the biodiversity changes within habitat recovery of Zostera noltii seagrass beds; and validate the dT-RFLP as a high-throughput tool to assess the system recovery. It was also proposed to develop a database of sequences related to individuals identified at species level to develop a new taxonomic reference system. A molecular phylogenetic analysis of the estuarine nematodes has being performed. After morphological identification, barcoding of 18S rDNA are being determined for each nematode species and the results have shown a good degree of concordance between traditional morphology-based identification and DNA sequences. The digest strategy developed for soil nematodes is not suitable for marine nematodes. Then five samples were cloned and sequenced and the sequence data was used to design a new dT-RFLP strategy to adapt this tool to marine assemblages. Several solutions were presented by DRAT and tested empirically to select the solution that cuts most efficiently, separating the different clusters. The results of quantitative PCR showed differences in nematode density between two sampling stations according the abundance of the nematode density obtained by the traditional methods. These results suggest that qPCR could be a robust tool for enumeration of nematode abundance, saving time.

Assessment of the Availability of Forest Biomass for Bioenergy Production in Southwest Portugal.

In 2014, Portugal was the seventh largest pellets producer in the World. Since the shortage of raw material is one of the major obstacles that the Portuguese sellets market faces, the need for a good assessment of biomass availability for energy purposes at both country and regional levels is reinforced. This work uses a Geographical Information System environment and remote sensing data to assess the availability and sustainability of forest biomass residues in a management unit with around 940 ha of maritime pine forest. The period considered goes from 2004 to 2015. The study area is located in Southwestern Portugal, close to a pellets factory; therefore the potential Contribution of the residual biomass generated in the management unit to the production of pellets is evaluated. An allometric function is used for the estimation of maritime pine above ground biomass. With this estimate, and considering several forest operations, the residual biomass available was assessed, according to stand composition and structure. This study shows that, when maritime pine forests are managed to produce wood, the amount of residues available for energy production is small (an average of 0.37 t ha -1  year -1 were generated in the study area between 2004 and 2015). As a contribution to the sustainability of the Portuguese pellets industries, new management models for maritime pine forests may be developed. The effect of the pinewood nematode on the availability of residual biomass can be clearly seen in this study. In the management unit considered, cuts were made to prevent dissemination of the disease. This contributes to a higher availability of forest residues in a specific period of time, but, in the medium term, they lead to a decrease in the amount of residues that can be used for energy purposes.