Essential oils as anti-nematode agents and their influence on in vitro nematode

Tese de doutoramento, Biologia (Biotecnologia), Universidade de Lisboa, Faculdade de Ciências, 2015Parasitic nematodes are among the most production-limiting plant pests. In Europe, the recent introduction of Bursaphelenchus xylophilus [the pinewood nematode (PWN)] and some species of Meloidogyne has proved damaging to forest ecosystems and crop production. Due to the very laborious and environment-dependent nature of greenhouse or field assays, in vitro host with parasite co-cultures can be a useful biotechnological tool to evaluate potential nematotoxic essential oils (EOs). The present work intended to a) screen EOs against PWN motility and M. chitwoodi [Columbia root-knot nematode (CRKN)] hatching, b) establish and characterize Pinus pinaster (maritime pine) in vitro cultures and Solanum tuberosum (potato) hairy roots (HR) cultures as well as P. pinaster with PWN and S. tuberosum HR with CRKN co-cultures, c) determine the effect of selected nematotoxic EOs on co-cultures growth or relative water content, nematode population and volatiles production. Ruta graveolens, Satureja montana and Thymbra capitata EOs revealed high activities against PWN. These and Dysphania ambrosioides and Filipendula ulmaria EOs also showed high inhibitory activity against CRKN. The established P. pinaster with PWN and S. tuberosum HR with CRKN co-cultures, displayed infection and developmental characteristics similar to those in nature. Of the most active EOs, those of R. graveolens and S. montana were chosen to be tested with in vitro co-cultures. S. montana EO was highly phytotoxic to both co-cultures, inhibiting potato HRs growth and inducing chlorosis and wilting in pine shoots. R. graveolens EO inhibited potato HRs with CRKN co-cultures growth but induced no macroscopic damages to in vitro P. pinaster with PWN co-cultures. In addition to constitutive compounds, biotransformation volatile compounds were detected after EOs addition to both co-cultures types. P. pinaster with PWN and S. tuberosum HR with CRKN co-cultures were a good system to mimic some of the natural infection conditions, allowing an overview of the combined host / parasite reactions, and being able to assist in the evaluation of EOs phytotoxicity and nematotoxic potential.Fundação para a Ciência e a Tecnologia (FCT), SFRH/BD/43738/2008, Pest-OE/EQB/LA0023/2011, PTDC/AGR-CFL/117026/201

Control of the root lesion Pratylenchus penetrans - the effect of nematocidal activity of plant-derived compounds

The root lesion nematode, Pratylenchus penetrans, is one of the most harmful plant parasites, responsible for worldwide productivity losses in a significant number of plant hosts. Generally, chemical control relies on synthetic compounds used through fumigation or direct contact, which offers a systemic protection. These control methods are costly and hazardous to the environment and to humans. Phytochemicals may play an important role in nematode control. The nematicidal activity of eight compounds that occur naturaly in plants, from two classes of compounds, was assessed at 2 mg/mL, for 24 h. Bioassays were performed following the standard direct contact methodology. P. penetrans was remarkably tolerant to the tested compounds, with mortality range between 1.0 and 5.8 %. To the best of our knowledge, the nematicidal activity of 4 phenolic compounds (catechin, caffeic acid, gallic acid and gentisic acid) was evaluated for the first time for P. penetrans

Plant-nematode co-cultures in the screening of sustainable nematicides against soil-dwelling parasitic nematodes of plants

The diseases caused by plant parasitic nematodes are still a serious constraint to modern global crop production. An increasing number of active compounds in commercial nematicidal formulations is being banned from use by common policies of pest management. Farmer communities report a low efficiency for the replacement pesticides, which reflects on crop yield and productivity. Novel sustainable biopesticides are urgently needed to cope with global food demands while respecting the most recent environmental policies. Plant-nematode co-cultures offer a stable biotechnological screening tool able to assess the active compound’s nematicidal activity and its effect on host tissues, simultaneously, in an easily accessible system that simulates natural infection. These systems are being developed and optimized at the Nematology laboratory of INIAV. Preliminary results were obtained for co-cultures of Solanum lycopersicum with the nematode Meloidogyne ethiopica and S. tuberosum with Globodera pallida. Future studies will target other plant parasitic nematodes, e.g., the root lesion nematodes (Pratylenchus penetrans) in transgenic roots of potato and the pinewood nematode (Bursaphelenchus xylophilus) on in vitro pine shoots (Pinus sp.)

Insights into the role of fungi in Pine Wilt Disease

Pine wilt disease (PWD) is a complex disease that severely affects the biodiversity and economy of Eurasian coniferous forests. Three factors are described as the main elements of the disease: the pinewood nematode (PWN) Bursaphelenchus xylophilus, the insect‐vector Monochamus spp., and the host tree, mainly Pinus spp. Nonetheless, other microbial interactors have also been considered. The study of mycoflora in PWD dates back the late seventies. Culturomic studies have revealed diverse fungal communities associated with all PWD key players, composed frequently of saprophytic fungi (i.e., Aspergillus, Fusarium, Trichoderma) but also of necrotrophic pathogens associated with bark beetles, such as ophiostomatoid or blue‐stain fungi. In particular, the ophiostomatoid fungi often recovered from wilted pine trees or insect pupal chambers/tunnels, are considered crucial for nematode multiplication and distribution in the host tree. Naturally occurring mycoflora, reported as possible biocontrol agents of the nematode, are also discussed in this review. This review discloses the contrasting effects of fungal communities in PWD and highlights promising fungal species as sources of PWD biocontrol in the framework of sustainable pest management actions

A review on the nematicidal activity of volatile allelochemicals against the pinewood nematode

The pinewood nematode (PWN), Bursaphelenchus xylophilus, induces shoot yellowing and wilting by injuring pine vascular tissues and resin canals. In Asia, it devastated wide forests of susceptible pine, causing drastic ecological, economic and cultural repercussions. In 1999, it reached Europe (Portugal) and despite the actions of the authorities, it rapidly advanced to the border areas of Spain, threatening the European pine forests. Chemical control is used in Asia with remarkable success, yet most nematicides are dangerous to human health and the environment. Natural volatile allelochemicals (VA) and derivatives are sustainable alternatives, providing many advantages to commercial nematicides. The present work summarizes available bibliographic information on VAs with activity against the PWN and reviews the chemical properties leading to nematoxicity. Published works reported over 250 VAs in direct contact assays. Compounds highly active against the PWN belong to monoterpenoid, phenylpropanoid and aliphatic alcohols, aldehydes, ketones, carboxylic acids, esters and sulphide groups. The presence of highly electronegative elements in these structures appears to increase activity. Trisulphides, coumarins, medium carbon chain length aliphatic alcohols and derivatives composed the top 5 most active allelochemicals. An indication of their mechanisms of action can be obtained from studying the reported structure-activity relationships. Keywords: Bursaphelenchus xylophilus; monoterpenoids; nematicide; oxygenated aliphatic compounds; phenylpropanoids; pinewood nematode; sulphides; toxicit

Toward an automated identification of Anastrepha fruit flies in the fraterculus group (Diptera, Tephritidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)In this study, we assess image analysis techniques as automatic identifiers of three Anastrepha species of quarantine importance, Anastrepha fraterculus (Wiedemann), Anastrepha obliqua (Macquart), and Anastrepha sororcula Zucchi, based on wing and aculeus images. The right wing and aculeus of 100 individuals of each species were mounted on microscope slides, and images were captured with a stereomicroscope and light microscope. For wing image analysis, we used the color descriptor Local Color Histogram; for aculei, we used the contour descriptor Edge Orientation Autocorrelogram. A Support Vector Machine classifier was used in the final stage of wing and aculeus classification. Very accurate species identifications were obtained based on wing and aculeus images, with average accuracies of 94 and 95%, respectively. These results are comparable to previous identification results based on morphometric techniques and to the results achieved by experienced entomologists. Wing and aculeus images produced equally accurate classifications, greatly facilitating the identification of these species. The proposed technique is therefore a promising option for separating these three closely related species in the fraterculus group.In this study, we assess image analysis techniques as automatic identifiers of three Anastrepha species of quarantine importance, Anastrepha fraterculus (Wiedemann), Anastrepha obliqua (Macquart), and Anastrepha sororcula Zucchi, based on wing and aculeus455554558CAPES - COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIORFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)09/54806-0; 14/16082-9sem informaçã

3D microsimulation of milkruns and pickers in warehouses using SIMIO

To help the Bosch Car Multimedia Portugal in Ferreiros, Braga to reduce its costs (both in time and space) with its warehouse, a micro simulation model is being developed in Simio. Particularly, the tool needs to be able to model pickers riding milkruns to collect containers of products, from a warehouse, to satisfy the needs of the production lines. In this sense, the storage strategy used on the warehouse, the quantity of requests a picker gets per shift, the time between shifts, the number of types of products, the arrival rate of requests, and the number of milkruns and pickers needs to be adjustable. Additionally, to design the corridors of the warehouse in a configurable way, an Add-in in C#, using the API of Simio, is being developed. Thus, this paper intends to document the first part of the simulation model developed, which consists on the pickers receiving requests and riding their milkruns to collect the respective containers from the warehouse. Five different Simio models compose the main simulation model. Conclusions and future work are discussed.This work has been co-supported by SI I&DT project in joint-promotion nº 36265/2013 (HMIEXCEL - 2013-2015 Project) and by FCT – Fundação para a Ciência e Tecnologia in the scope of the project: PEst-OE/EEI/UI0319/2014

Mechanical pruning and soil organic amending in two terroirs. Effects on wine chemical composition and sensory profile

The knowledge about the interaction between mechanical pruning and soil organic amending is still scarce. This study aimed to examine the effects of the interaction between these two practices on wine quality. Syrah grapes from two trial fields in Portugal subjected to two different pruning systems (mechanical pruning; hand spur pruning) and five different organic amendment treatments (control, biochar, municipal solid waste compost, cattle manure, and sewage sludge) were harvested and vinified for four years. Mechanical pruning significantly reduced wine alcoholic strength, pH, and total anthocyanins. Mechanical pruning and organic amendments, tendentially reduced wine total phenols and tannin power, known as an “estimation of the astringency potential of the wines”. Tasters found low but significant differences in global appreciation with the pruning system. Sludge tended to reduce wine global appreciation more than municipal solid waste compost and cattle manure, while biochar had no effect on tasters’ preference when compared to the control. There was strong relation between yield and tasters’ preference only above 6 kg/vine and 8 kg/vine depending on the terroir. Mechanical pruning tendentially has significant effects on wine quality when yield raises above a certain level. Thus, with this pruning system, the choice of the organic amendment and its amount must be done considering the destiny of the produced grapes. To the best of our knowledge, effects of the interaction of mechanical pruning with soil organic amending on wine quality are a noveltyinfo:eu-repo/semantics/publishedVersio

How parasitism genes are regulated: a motif to search for genes regulators in the plant parasitic nematode Bursaphelenchus xylophilus

Plant-parasitic nematodes threaten global agricultural and forestry systems. The search for new control strategies in line with the EU’s sustainability goals highlight significant knowledge gaps. Like all other plant pathogens, plant-parasitic nematodes deliver several parasitism proteins (effectors) into the host plant to cause disease. NemaWAARS project focuses on mechanism(s) of regulation and gene control expression of parasitism genes in pinewood nematode, Bursaphelenchus xylophilus. From the previous transcriptomic data derived from the pharyngeal gland cells (considered a specialized tissue potentially related to parasitism) we have identified a non-coding DNA motif - STATAWAARS - associated in the promotor region of highly abundant and secreted expressed genes. Given that this non-coding genetic signature unifies many sequences of unrelated parasitism genes, it implies the existence of a potential major regulator(s), that binds to this sequence to control the expression of downstream genes. We hypothesize that by disrupting this regulator(s), it would be possible to simultaneously disrupt the expression of many associated parasitism-related genes. To test the hypothesis the project aims to identify proteins (or complex of proteins) that bind in the promoter regions of parasitism-related genes (in vivo) or identify other regulatory candidates for master regulators of parasitism-related genes expression that are enriched in the pharyngeal gland cell tissues. For the best candidate regulatory proteins, an RNAi approach will target the selected gene candidates and evaluate the regulatory role in effector genes expression and in interaction with the host (in planta). Under an ongoing national and international collaborative network, the strategy in NemaWAARS will include innovative approaches to explore the regulators that govern effector gene expression applied in B. xylophilus research

Atlantic mammal traits: a dataset of morphological traits of mammals in the atlantic forest of south America

Measures of traits are the basis of functional biological diversity. Numerous works consider mean species-level measures of traits while ignoring individual variance within species. However, there is a large amount of variation within species and it is increasingly apparent that it is important to consider trait variation not only between species, but also within species. Mammals are an interesting group for investigating trait-based approaches because they play diverse and important ecological functions (e.g., pollination, seed dispersal, predation, grazing) that are correlated with functional traits. Here we compile a data set comprising morphological and life history information of 279 mammal species from 39,850 individuals of 388 populations ranging from −5.83 to −29.75 decimal degrees of latitude and −34.82 to −56.73 decimal degrees of longitude in the Atlantic forest of South America. We present trait information from 16,840 individuals of 181 species of non-volant mammals (Rodentia, Didelphimorphia, Carnivora, Primates, Cingulata, Artiodactyla, Pilosa, Lagomorpha, Perissodactyla) and from 23,010 individuals of 98 species of volant mammals (Chiroptera). The traits reported include body mass, age, sex, reproductive stage, as well as the geographic coordinates of sampling for all taxa. Moreover, we gathered information on forearm length for bats and body length and tail length for rodents and marsupials. No copyright restrictions are associated with the use of this data set. Please cite this data paper when the data are used in publications. We also request that researchers and teachers inform us of how they are using the data.Fil: Gonçalves, Fernando. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Bovendorp, Ricardo S.. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Beca, Gabrielle. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Bello, Carolina. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Costa Pereira, Raul. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Muylaert, Renata L.. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Rodarte, Raisa R.. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Villar, Nacho. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Souza, Rafael. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Graipel, Maurício E.. Universidade Federal de Santa Catarina; BrasilFil: Cherem, Jorge J.. Caipora Cooperativa, Florianopolis; BrasilFil: Faria, Deborah. Universidade Estadual de Santa Cruz; BrasilFil: Baumgarten, Julio. Universidade Estadual de Santa Cruz; BrasilFil: Alvarez, Martín R.. Universidade Estadual de Santa Cruz; BrasilFil: Vieira, Emerson M.. Universidade do Brasília; BrasilFil: Cáceres, Nilton. Universidade Federal de Santa María. Santa María; BrasilFil: Pardini, Renata. Universidade de Sao Paulo; BrasilFil: Leite, Yuri L. R.. Universidade Federal do Espírito Santo; BrasilFil: Costa, Leonora Pires. Universidade Federal do Espírito Santo; BrasilFil: Mello, Marco Aurelio Ribeiro. Universidade Federal de Minas Gerais; BrasilFil: Fischer, Erich. Universidade Federal do Mato Grosso do Sul; BrasilFil: Passos, Fernando C.. Universidade Federal do Paraná; BrasilFil: Varzinczak, Luiz H.. Universidade Federal do Paraná; BrasilFil: Prevedello, Jayme A.. Universidade do Estado de Rio do Janeiro; BrasilFil: Cruz-Neto, Ariovaldo P.. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Carvalho, Fernando. Universidade do Extremo Sul Catarinense; BrasilFil: Reis Percequillo, Alexandre. Universidade de Sao Paulo; BrasilFil: Paviolo, Agustin Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú | Universidad Nacional de Misiones. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú; ArgentinaFil: Duarte, José M. B.. Universidade Estadual Paulista Julio de Mesquita Filho; Brasil. Fundación Oswaldo Cruz; BrasilFil: Bernard, Enrico. Universidade Federal de Pernambuco; BrasilFil: Agostini, Ilaria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú | Universidad Nacional de Misiones. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú; ArgentinaFil: Lamattina, Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Ministerio de Salud de la Nación; ArgentinaFil: Vanderhoeven, Ezequiel Andres. Ministerio de Salud de la Nación; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentin