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The rice root-knot nematode (RRKN), Meloidogyne graminicola Golden and Birchfield 1965, is a dangerous crop pest that affects rice production on a global scale. The largest rice-producing countries struggle with the impacts of RRKN infestation, namely, underdeveloped plants and a reduction in rice grain that can reach up to 70% of crop yield. In addition, the shift to strategies of sustainable pest management is leading to a withdrawal of some of the most effective pesticides, given the dangers they pose to the environment and human health. Volatile metabolites produced by plants can offer safer alternatives. The present study characterized the nematicidal activity of volatile phytochemicals against the RRKN and compared the most active with commercial nematicides concerning their safety to the environment and human health. Rice plants were used to grow large numbers of RRKNs for direct-contact bioassays. Mortality induced by the volatiles was followed for four days on RRKN second-stage juveniles. Of the 18 volatiles tested, carvacrol, eugenol, geraniol, and methyl salicylate showed the highest mortalities (100%) and were compared to traditional nematicides using (eco)toxicological parameters reported on freely available databases. While methyl salicylate had a faster activity, carvacrol had more lasting effects. When compared to synthetic nematicides, these volatile phytochemicals were reported to have higher thresholds of toxicity and beneficial ecotoxicological parameters. Ultimately, finding safer alternatives to traditional pesticides can lower the use of damaging chemicals in farming and leverage the transition to agricultural practices with a lower impact on biodiversity. [ABSTRACT FROM AUTHOR]

Pines are the most economically important trees in the world and, together with eucalyptus, they dominate commercial forests. But the success of a relatively small number of widely planted species, such as Pinus pinaster, the maritime pine, comes at a price. Pines are attractive to damaging pathogens and insect pests, including the pinewood nematode (PWN), Bursaphelenchus xylophilus, the causal agent of pine wilt disease (PWD). Originally described in Japan, PWD has caused widespread destruction to forests in countries such as China, Taiwan, Portugal, Spain and the United States. PWN causes irreparable damage to the vascular system of its pine hosts, leading to mortality within 3 months. Pine sawyer beetles (Monochamus spp.) are key vectors of PWD, introducing the PWN to healthy trees during feeding. Other organisms contribute to PWD spread and development, including bacteria, fungi and bark beetles. Control measures include tree felling to prevent vector transmission of PWN, insecticide treatments, trapping of Monochamus spp. and tree breeding for plant resistance. The PWN is a quarantine pathogen and subject to regular legislation and phytosanitary measures aimed at restricting movement and preventing introduction to new areas. Current research is investigating the use of biopesticides against PWN and Monochamus spp. This review examines the biology, epidemiology, impact and management of PWD through published research, grey literature and interviews with people directly involved in the management of the disease in Portugal. [ABSTRACT FROM AUTHOR]

Numerous plastic products are used in agriculture, including containers, packaging, tunnels, drip irrigation tubing, and mulches. Large amounts of plastics are used as mulches on the soil surface for vegetable and fruit production (tomato, cucumber, watermelon, strawberry, and vine) to reduce weed competition, increase water and fertilizer use efficiency, and enhance crop yield. Portugal uses around 4500 t/year of polyethylene to cover approximately 23,000 ha of agricultural land, and only a small amount is recovered for recycling or secondary uses because of issues of contamination with the soil, vegetation, pesticides, and fertilizers. Cleaning and decontaminating polyethylene mulch are costly, and commercial technology is often not accessible or economical. Most plastic mulch is composed of polyethylene that degrades slowly and produces a large quantity of residues in the soil, with a negative impact on the environment. In the present study, the effects of long-term cultivation of blueberry using green 100% high-density polyethylene mulch in the south Portugal were evaluated for soil chemical and biological changes. High-density green plastic mulch did not contaminate the topsoil with di(2-ethylhexyl) phthalate, and heavy metals, buttotal nitrogen, organic carbon concentrations, electric conductivity, and microbial activity were significantly reduced in the planting row compared with the bare soil without mulching. Furthermore, the presence of plastic mulch did not negatively affect the presence of nematodes, and the number of Rhabditida (bacterial feeders) increased in the planting and covered row. [ABSTRACT FROM AUTHOR]

Considered one of the most devastating plant parasitic nematodes worldwide, Meloidogyne spp. (commonly known as the root-knot nematodes (RKNs)) are obligate sedentary endoparasites that establish in the roots, causing hyperplasia and hypertrophy of surrounding cells, triggering the formation of galls. These galls will affect root development and physiology, leading to substantial yield losses. During 2017–2022, an extensive survey of Meloidogyne species was undertaken in Portugal (mainland and islands). A total of 1,071 samples were collected by the National Plant Protection Organization (DGAV) and private farmers from different regions of the country and were analysed at the Laboratory of Nematology (NemaINIAV). Samples in which the presence of Meloidogyne sp. was detected were used to perform bioassays to obtain females and juveniles for further studies. Since the accurate identification of RKNs is an important aspect of crop management, morphological and biochemical characterisation was performed. The most common morphological features were observed, showing consistency with previous descriptions of the genus. The biochemical identification using the esterase (EST) phenotype revealed the phenotypes of Meloidogyne arenaria, M enterolobi, M. hispanica, M. hapla, M. incognita, M javanica, and M. luci. Meloidogyne incognita and M. javanica were found to be the most prevalent species in the different regions followed by M. arenaria and M. hapla. This is the first distribution report performed in Portugal on RKNs, contributing to the development of management strategies and to updated information on the status of these pests in Europe. [ABSTRACT FROM AUTHOR]

Soil health is intimately intertwined with ecosystem services. Climate change negatively impacts ecosystem functioning, by altering carbon and nitrogen biogeochemical cycles and shifting nutrient bioavailability, thus hampering food production and exacerbating biodiversity loss. Soil ecosystem services are provided by belowground biota, and as the most abundant metazoans on Earth, nematodes are key elements of soil food webs and reliable bioindicators of soil health. Here, we carry out a literature review from 2019, the year that the Intergovernmental Panel on Climate Change published a report relating and expressing serious concerns on the effects of climate change on the land degradation and sustainability of terrestrial ecosystems. We focus on documenting and discussing the composition of nematode communities contributing to improving soil health, and soil management practices to promote their presence and limit the effects of climate change on soils. By recognizing beneficial nematodes as plant-promoting agents, we could harness their potential to our benefit, catalyze decomposition services, improve plant performance, and increase carbon sequestration. This way, we will contribute to soil health and a well-balanced and well-managed system, making it possible to increase productivity, guarantee food security, and reduce the yield gap, with a limited human footprint on the environment. [ABSTRACT FROM AUTHOR]

Climate change is prompting a shift of tropical pests to locations with a previously lower probability of invasion. This is the case for root-knot nematodes, Meloidogyne sp., particularly of the tropical group. Among them, M. ethiopica is now considered a threat to European food security. The development of novel sustainable nematicides can be based on in vitro bioassays of highly active phytochemicals, e.g., volatiles from essential oils. However, a steady supply of nematodes is often very difficult and dependent on environmental conditions. In the present study, an in vitro co-culture system of M. ethiopica parasitizing hairy roots of Solanum lycopersicum was established, for the first time, to easily obtain populations of second-stage juveniles (J2). These were then used to screen the nematicidal activity of 10 volatile compounds characteristic of essential oils. Finally, information on the most successful compounds was reviewed to predict their environmental dispersion and ecotoxicological hazards. The M. ethiopica population obtained from the co-culture was morphologically similar to reported populations in natural conditions and could be accurately used in direct-contact bioassays. The aldehydes citral and citronellal induced complete mortality of the tested J2, at 1 mg/mL, while compounds from other chemical groups were not as successful. In comparison to commonly used commercial nematicides, citral and citronellal were less likely to accumulate in the water environmental compartment and have lower reported toxicities compared to aquatic organisms and to mammals. Overall, in vitro co-cultures showed the potential to expedite the screening and discovery of bioactive compounds as a contribution to the development of sustainable biopesticides, as well as to lower the impacts of modern farming on agroecosystems. [ABSTRACT FROM AUTHOR]

Control of the pinewood nematode (PWN), the causal agent of pine wilt disease, can be achieved through the trunk injection of nematicides; however, many pesticides have been linked to environmental and human health concerns. Essential oils (EOs) are suitable alternatives due to their biodegradability and low toxicity to mammals. These complex mixtures of plant volatiles often display multiple biological activities and synergistic interactions between their compounds. The present work profiled the toxicity of eight EOs against the PWN in comparison to their 1:1 mixtures, to screen for successful synergistic interactions. Additionally, the main compounds of the most synergistic mixtures were characterized for their predicted environmental fate and toxicity to mammals in comparison to emamectin benzoate, a commercial nematicide used against PWN. The mixtures of Cymbopogon citratus with Mentha piperita and of Foeniculum vulgare with Satureja montana EOs showed the highest activities, with half-maximal effective concentrations (EC50) of 0.09 and 0.05 µL/mL, respectively. For these, complete PWN mortality was reached after only ca. 15 min or 2 h of direct contact, respectively. Their major compounds had a higher predicted affinity to air and water environmental compartments and are reported to have very low toxicity to mammals, with low acute oral and dermal toxicities. In comparison, emamectin benzoate showed lower nematicidal activity, a higher affinity to the soil and sediments environmental compartments and higher reported oral and dermal toxicity to mammals. Overall, uncovering synergistic activities in combinations of EOs from plants of different families may prove to be a source of biopesticides with optimized toxicity against PWNs. [ABSTRACT FROM AUTHOR]

The potato cyst nematode (PCN), Globodera pallida, has acquired significant importance throughout Europe due to its widespread prevalence and negative effects on potato production. Thus, rapid and reliable diagnosis of PCN is critical during surveillance programs and for the implementation of control measures. The development of innovative technologies to overcome the limitations of current methodologies in achieving early detection is needed. Lab-on-a-chip devices can swiftly and accurately detect the presence of certain nucleotide sequences with high sensitivity and convert the presence of biological components into an understandable electrical signal by combining biosensors with microfluidics-based biochemical analysis. In this study, a specific DNA-probe sequence and PCR primers were designed to be used in a magnetoresistive biosensing platform to amplify the internal transcribed spacer region of the ribosomal DNA of G. pallida. Magnetic nanoparticles were used as the labelling agents of asymmetric PCR product through biotin–streptavidin interaction. Upon target hybridization to sensor immobilized oligo probes, the fringe field created by the magnetic nanoparticles produces a variation in the sensor's electrical resistance. The detection signal corresponds to the concentration of target molecules present in the sample. The results demonstrate the suitability of the magnetic biosensor to detect PCR target product and the specificity of the probe, which consistently distinguishes G. pallida (DV/V > 1%) from other cyst nematodes (DV/V < 1%), even when DNA mixtures were tested at different concentrations. This shows the magnetic biosensor's potential as a bioanalytical device for field applications and border phytosanitary inspections. [ABSTRACT FROM AUTHOR]

Simple Summary: Damage caused by plant-parasitic nematodes (PPNs) is usually underestimated as many farmers are oblivious of their presence. Root-knot nematodes (RKNs) are an economically important group of PPNs and one of the most devastating. The species Meloidogyne arenaria, M. hapla, M. incognita and M. javanica are considered the most important, due to their worldwide distribution and polyphagia, being the northern RKN, Meloidogyne hapla the most important species occurring in cold regions. During 2019–2022, surveys were carried out in different districts of Portugal in horticultural and ornamental crops. From the samples collected, nine were identified morphologically and molecularly as M. hapla, indicating that its prevalence is increasing, and demonstrating its importance and impact in agricultural systems. Root-knot nematodes (RKNs), Meloidogyne spp., are a group of plant-parasitic nematodes (PPNs) of great economic significance worldwide. The northern root-knot nematode, Meloidogyne hapla, is one of the most important species of RKNs occurring in cold regions. In Portugal so far, M. hapla has been found parasitizing potato and fig trees. During surveys carried out in 2019–2022 in fields for horticultural and ornamental production, soil and root samples were collected. Roots were observed under a stereomicroscope to determine the presence of galls. Nematodes were extracted from the soil. Morphological features showed a high similarity and consistency with previous descriptions of the genus. For molecular analysis, total genomic DNA was isolated from single nematodes and used to amplify in multiplex reaction using the species-specific primers JMV1, JMV2 and JMVhapla, and for sequencing of the ITS region with the primers TW81/AB28. Multiplex PCR amplification produced a specific fragment of 440 bp and PCR amplification of the ITS region yielded a single fragment of 550 bp, as expected. The obtained sequences showed a similarity ranging from 99.8% to 100% with the sequences of M. hapla available in the database. The phylogenetic tree revealed that the isolates grouped with M. hapla isolates. From the 690 samples collected, M. hapla was detected in three different hosts (grapevine, eucalyptus and potato) in four districts of mainland Portugal and on Madeira Island. To our knowledge, this is the first report of M. hapla infecting the grapevine and eucalyptus in Portugal. [ABSTRACT FROM AUTHOR]

Plant-parasitic nematodes (PPNs) are among the most notorious and underrated threats to food security and plant health worldwide, compromising crop yields and causing billions of dollars of losses annually. Chemical control strategies rely heavily on synthetic chemical nematicides to reduce PPN population densities, but their use is being progressively restricted due to environmental and human health concerns, so alternative control methods are urgently needed. Here, we review the potential of bacterial and fungal agents to suppress the most important PPNs, namely Aphelenchoides besseyi, Bursaphelenchus xylophilus, Ditylenchus dipsaci, Globodera spp., Heterodera spp., Meloidogyne spp., Nacobbus aberrans, Pratylenchus spp., Radopholus similis, Rotylenchulus reniformis, and Xiphinema index. [ABSTRACT FROM AUTHOR]

Considered one of the most devastating plant–parasitic nematodes worldwide, Bursaphelenchus xylophilus (commonly known as pinewood nematode, PWN) is the causal agent of the pine wilt disease in the Eurasian coniferous forests. This migratory parasitic nematode is carried by an insect vector (Monochamus spp.) into the host tree (Pinus species), where it can feed on parenchymal cells and reproduce massively, resulting in the tree wilting. In declining trees, PWN populations are strongly dependent on fungal communities colonizing the host (predominantly ophiostomatoid fungi known to cause sapwood blue-staining, the blue-stain fungi), which not only influence their development and life cycle but also the number of individuals carried by the insect vector into a new host. Our main aim is to understand if PWN-associated mycobiota plays a key role in the development of PWD, in interaction with the PWN and the insect vector, and to what extent it can be targeted to disrupt the disease cycle. For this purpose, we characterized the fungal communities of Pinus pinaster trees infected and non-infected with PWN in three collection sites in Continental Portugal with different PWD temporal incidences. Our results showed that non-infected P. pinaster mycoflora is more diverse (in terms of abundance and fungal richness) than PWN-infected pine trees in the most recent PWD foci, as opposed to the fungal communities of long-term PWD history sites. Then, due to their ecological importance for PWN survival, representatives of the main ophiostomatoid fungi isolated (Ophiostoma, Leptographium , and Graphilbum) were characterized for their adaptative response to temperature, competition in-between taxa, and as food source for PWN. Under the conditions studied, Leptographium isolates showed promising results for PWN control. They could outcompete the other species, especially O. ips , and significantly reduce the development of PWN populations when compared to Botrytis cinerea (routinely used for PWN lab culturing), suggesting this to be a natural antagonist not only for the other blue-stain species but also for the PWN. [ABSTRACT FROM AUTHOR]

Simple Summary: The presence of the oak pinhole borer, the insect Platypus cylindrus, in Portuguese cork oak stands has drastically increased in the past few decades. This beetle excavates long galleries in the trunk while inoculating fungi (called ambrosia fungi) transported in special organs (mycangia) that will serve as food source for its offspring. The combined action of extensive boring into the heartwood and the inoculation of fungi leads to an increase in tree mortality. A new ambrosial fungus, Ceratocystiopsis quercina, was isolated from the insects' mycangia and from wilting trees, namely from the staining patches it causes on wood. Platypus cylindrus is the most common ambrosia beetle in stands of Quercus suber in Portugal. This insect farms specialized fungi in sapwood galleries, using its mycangia to carry and store these organisms. Some ectosymbiotic fungi carried by P. cylindrus are phytopathogenic and cause extensive tree mortality and severe economic losses. To understand the role of P. cylindrus fungal symbionts in stands of Q. suber we examined beetle galleries present in declining and/or dying cork oak trees during field surveys. Logs with active galleries were obtained in situ and from captured emerging beetles. Insects were aseptically dissected, and their mycangia and intestine were retrieved. Morphological and molecular profiles of fungal isolates obtained from cultured insect parts were carried out to accurately characterize and identify isolated fungi. Molecular characterizations were performed with DNA sequence data from four loci, i.e., LSU, SSU, 5.8S-ITS2-28S, and TUB. Morphological results consistently showed a collection of Ophiostoma-like fungal axenic isolates, while phylogenies inferred that this collection constitutes an undescribed taxon reported herein for the first time in association with P. cylindrus in Portuguese cork oak stands. The novel species was erected as Ceratocystiopsis quercina sp. nov. and constitutes a new phytopathogenic fungal species associated with symptoms of vegetative cork oak decline. [ABSTRACT FROM AUTHOR]

Pine wilt disease (PWD), caused by the pinewood nematode (PWN) Bursaphelenchus xylophilus, is one of the most destructive diseases in trees of the genus Pinus and is responsible for significant environmental and economic losses in North America, Eastern Asia, and Western Europe. However, pine species are not equally affected, with some being tolerant/resistant while others are susceptible to nematode infection for reasons still unclear. The present study aims to investigate differential chemical responses of susceptible and tolerant/resistant pine species shortly after nematode infection by characterizing the phenolic profiles of adult Pinus sylvestris, Pinus pinaster, Pinus pinea, and Pinus halepensis. HPLC and LC-MS were used to identify and quantify the pine´s phenolic compounds: gallic acid, ferulic acid, taxifolin, rutin, resveratrol, (+)-secoisolariciresinol, (−)-epicatechin, protocatechuic acid hexoside, gallic acid hexoside, ferulic acid glucoside, quercetin hexoside, and two unidentified compounds (#A and #B). Prior to infection, we could not differentiate between nematode-tolerant/resistant and susceptible adult pine species based on their constitutive phenolic compounds. In the presence of the PWN, the phenolic profile allowed for a noticeable separation of the PWN-tolerant/resistant P. halepensis from the susceptible P. sylvestris, contrasting with a more homogenous response from P. pinea and P. pinaster. Observations on P. halepensis suggest that taxifolin, resveratrol, and rutin may have an active role in protecting against B. xylophilus, possibly in conjugation with other biochemical and anatomical characters. We emphasize the importance of studying pine tolerant/resistance on adult trees, and not on excised branches, saplings, or seedlings to accurately simulate the nematode–pine host interactions occurring under natural conditions. [ABSTRACT FROM AUTHOR]

Pin nematodes (Paratylenchus spp.) currently comprise 132 species of polyphagous plant ectoparasites with at least seven species that are plant pathogenic emphasizing the need for correct identification to establish an appropriate management strategy. Sequences of highly conserved regions of ribosomal and mitochondrial RNA genes are a powerful species-level diagnostic tool within Tylenchulidae. A nematological survey was conducted from 2019 to 2021 in commercial vineyards distributed in four major wine-producing regions in the Central and South Portugal. Nine populations of Paratylenchus spp. were isolated from the rhizosphere of grapevines that were characterized from morphological data and molecular phylogenetic analysis using two rRNA genes (D2–D3 expansion segments of the 28S, and partial 18S) and a region partial of the COI mtRNA gene. Contrasting morphological hypotheses with molecular data provided rapid detection of six species, specifically P. goodeyi, P. hamatus, P. pedrami, P. tenicaudatus, P. variabilis, and P. veruculatus. Bayesian inference (BI) phylogenetic trees for these molecular markers established phylogenetic relationships underscore the importance of using genomic and molecular phylogenetic data for accurate pin nematode identification at the species level. To our knowledge, this is the first report of Paratylenchus spp. in Portugal, extending the geographical distribution of these species in the Mediterranean Basin, and the first record of P. goodeyi, P. pedrami, P.variabilis, P. veruculatus and P. tenicaudatus parasitizing grapevine. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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Esteya vermicola is a potential biological agent of the pinewood nematode (PWN), Bursaphelenchus xylophilus, due to its high infectivity. However, knowledge of E. vermicola colonization in the host pine tree is less known. To reveal the distribution pattern of E. vermicola inside the pine tree and the interactions between fungus and PWN, biocontrol tests were conducted on 10 years old Pinus densiflora. A green fluorescence protein (GFP)‐tagged E. vermicola was used to observe the fungal hyphae. Real‐time TaqMan PCR quantification was applied to quantify the fungal hyphae in host tree. The results suggest that inoculation with the fungus significantly improved the survival rate of tested trees. Besides, the number of PWN extracted from fungal inoculated tested trees significantly decreased. Moreover, the fungal hyphae and lunate conidia grown in tested trees were observed with the GFP‐tagged E. vermicola. The real‐time quantification of E. vermicola illustrated the differential distribution of E. vermicola in wilted, wilting and healthy pine trees. Additionally, the promotion of fungal hyphal dispersion by B. xylophilus was found in the pine trees artificially infected with PWN. This study reveals the distribution of E. vermicola and PWN during biocontrol of pine wilt disease and provides direct evidence of the effectiveness of this fungus to control PWN. The migration of PWN infected by E. vermicola was supposed to improve the fungal extension in host pine tree. [ABSTRACT FROM AUTHOR]

Six species of the genus Bursaphelenchus were recovered as the result of a survey to assess the potential introduction of Bursaphelenchus xylophilus into the conifer forests of Romania. Bursaphelenchus xylophilus was not detected in the sampling regions. Bursaphelenchus dietrichi was recovered from one point, representing the first report of this species in the country. The other detected species, B. abietinus, B. andrassyi, B. hofmanni, B. fraudulentus and B. willibaldi, were characterized by their morphometric data and molecular analyses. The morphometrics of Romanian populations of B. hofmanni and B. willibaldi are presented for the first time. The phylogenetic relationships of the recovered species (except for B. abietinus, with no successful sequencing) were reconstructed using the sequences of the D2‐D3 expansion segments of the large subunit of ribosomal DNA (D2‐D3LSU rDNA). Some of the species were also characterized using the restriction fragment length polymorphism (RFLP) pattern of the internal transcribed spacer (ITS) fragment. The distribution, related host trees and phylogenetic relationships of the identified species are discussed. [ABSTRACT FROM AUTHOR]

As a result of the detection of the pinewood nematode Bursaphelenchus xylophilus in Portugal, and its subsequent spread to Spain, intense surveys were conducted to screen for the presence of Bursaphelenchus species in Romania. Herein, we report recent surveys of insects potentially vectoring Bursaphelenchus species collected using trap trees or pheromone‐baited traps placed in the forest. Trap felled spruce trees (Picea abies) and pheromone‐baited traps were installed in six different counties in Romania (Brașov, Sibiu, Suceava, Hunedoara, Timiș and Dâmbovița). Ten different species of insects distributed among Curculionidae and Cerambycidae were obtained. Nematodes were extracted from insects and observed to validate the presence of Bursaphelenchus specimens. One female identified as Monochamus sutor was the only specimen carrying nematodes in the genus Bursaphelenchus. Nematodes were identified as B. mucronatus based on morphological and molecular features. This is the first detection and report of natural spread of B. mucronatus in Romania. The absence of B. xylophilus was confirmed in the areas of Romania surveyed in this work. [ABSTRACT FROM AUTHOR]

Context Pine wilt disease is caused by the pinewood nematode (Bursaphelenchus xylophilus (Steiner et Buhren) Nickle), a quarantine pest, and is a concern to maritime pine (Pinus pinaster) in Portugal due to its economic, environmental, and social impacts. This disease is regarded as a major threat to European forests. Aims This paper aimed to evaluate the genetic variation in maritime pine families that were inoculated with pinewood nematode, identify the most resistant families, and establish the guidelines for a resistance improvement program. Methods Two-year-old half-sib progenies obtained from 96 plus trees were inoculated. The plants were monitored for survival on four different dates. The statistical analysis followed the mixed model theory. Results Genetic variability of the susceptibility to pine wilt disease was observed. At 157 days after inoculation, the 15 highest genetic ranking families out of 96 were selected, having a predicted survival mean of 15.6% instead of 11.0% on average for the all 96 families. Conclusion This study allows for the implementation of an improvement program to help control pine wilt disease. [ABSTRACT FROM AUTHOR]

The potato cyst nematodes (PCN) Globodera rostochiensis and G. pallida pose one of the greatest threats to potato crops worldwide and are subject to strict quarantine regulations in many countries. The identification of these Globodera species based on morphology may be ambiguous due to the variability of the main morphological features and the overlapping of the standard parameters in these two species; thus, confirmation via molecular methods is recommended. Multiplex PCR with speciesspecific primers (ITS5/PITSp4 + PITSr3) allows both species to be distinguished. However, despite the development of molecular identification methods, the morphological approach remains useful as a complementary diagnostic technique. In this work, we report results of morphological and molecular analyses that were carried out in two Globodera species from Portuguese potato fields. The average morphometric values of 40 cysts and 40 second-stage juveniles were generally within the expected ranges for G. pallida and G. rostochiensis with some variations noted. Molecular analysis with multiplex PCR confirmed the morphometric identification. The present results confirmed the occurrence of two potato cyst nematode species, G. rostochiensis and G. pallida. Surprisingly, the analysis of soils from Portuguese potato fields detected a greater number of samples infested with G. pallida, which is contrary to expectation as G. rostochiensis has been considered the most widespread species in Portugal. The distinction between the two species is therefore essential in order to detect their presence in the country with a view to re-evaluating the control measures implemented so far and adopting more effective practices. [ABSTRACT FROM AUTHOR]

Background The pinewood nematode ( PWN) Bursaphelenchus xylophilus is an important conifer disease worldwide. It is the direct cause of the death of millions of pines in south-east Asia (mainly Japan, China and Korea) and has been established in Portugal since 1999. The phasing out of methyl bromide has created an urgent need for alternative treatment of wood packaging materials. The effect of sulfuryl fluoride ( SF), a broad-spectrum fumigant used to control insects, was tested in Pinus pinaster boards naturally infested by PWN. Results Boards were fumigated for 24 h at three different temperatures (15, 20 and 30 °C) with dosage ranges of 3169-4407, 1901-4051 and 1385-2141 gh m−3 respectively. Treated wood was sampled for nematode identification and counting, before treatment and after 24 h, 72 h and 21 days. No survival was found in the 15 °C and 30 °C treatments, while at 20 °C the mortality ranged from 94.06 to 100%. Some reasons for the survival at 20 °C are presented. Conclusion Results confirm SF to be an effective quarantine treatment for PWN at 15 and 30 °C. Further studies are needed to obtain the most effective dosage at 20 °C, and to determine the toxicity of SF fumigation on B. xylophilus at other temperatures, especially at 25 °C. © 2012 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

The pinewood nematode (PWN), Bursaphelenchus xylophilus, is the causal agent of pine wilt disease (PWD) and a quarantine organism in many countries. Managing PWD involves strict regulations and heavy contingency plans, and present climate change scenarios predict a spread of the disease. The urgent need for sustainable management strategies has led to an increasing interest in promising biocontrol agents capable of suppressing the PWN, like endoparasitic nematophagous fungi of the Esteya genus. Here, we review different aspects of the biology and ecology of these nematophagous fungi and provide future prospects. [ABSTRACT FROM AUTHOR]

Simple Summary: New risks to plant health are constantly emerging. Such is the case of the rice root knot nematode Meloidogyne graminicola, adapted to flooded conditions and representing a risk to all types of rice agro-systems. It has been recently detected in Italy and added to the European and Mediterranean Plant Protection Organization (EPPO) Alert List. The presence of this nematode in Europe poses a threat to rice production, as there is a high probability to spread, due to trade activities and climate changes. In view of its importance, an extensive updated review was carried out. Rice (Oryza sativa L.) is one of the main cultivated crops worldwide and represents a staple food for more than half of the world population. Root-knot nematodes (RKNs), Meloidogyne spp., and particularly M. graminicola, are serious pests of rice, being, probably, the most economically important plant-parasitic nematode in this crop. M. graminicola is an obligate sedentary endoparasite adapted to flooded conditions. Until recently, M. graminicola was present mainly in irrigated rice fields in Asia, parts of the Americas, and South Africa. However, in July 2016, it was found in northern Italy in the Piedmont region and in May 2018 in the Lombardy region in the province of Pavia. Following the first detection in the EPPO region, this pest was included in the EPPO Alert List as its wide host range and ability to survive during long periods in environments with low oxygen content, represent a threat for rice production in the European Union. Considering the impact of this nematode on agriculture, a literature review focusing on M. graminicola distribution, biology, identification, and management was conducted. [ABSTRACT FROM AUTHOR]

The potato cyst nematode (PCN) Globodera pallida has acquired significant importance throughout Europe due to its nefarious effects on potato production. Rapid and reliable diagnosis of PCN is critical during the surveillance programs and for the implementation of control measures. Molecular DNA-based methods are available, but they require expensive laboratory facilities, equipment and trained technicians. Moreover, there is an additional need of time for sample shipment and testing. In this work, we have developed a new and simple assay which reliably discriminates G. pallida from other cyst nematodes in less than 40 min. This assay may be applied either on cysts or juveniles with the ability to detect a single juvenile of G. pallida in a sample of at least 40 juveniles of the non-target species G. rostochiensis. This test should be a tool to improve the performance of the laboratory and has the potential to be performed on-site. [ABSTRACT FROM AUTHOR]

Research Highlights: Raffaelea quercina sp. nov. is an ophiostomatoid fungus isolated from the ambrosia beetle Platypus cylindrus. The species occurs in symptomatic Portuguese cork oak trees, (Quercus suber L.), exhibiting vegetative decline. Background and Objectives: Quercus suber L. is a species restricted to the Mediterranean basin, of special economic importance as it constitutes the crucial raw material for the cork production industry, in particular for Portugal, the world's leading producer. Over the last three decades a progressive and alarming decline of cork oak trees has been observed across its distribution area, including Portugal. The ambrosia beetle Platypus cylindrus, commonly known as the oak pinhole borer, establishes symbiotic relationships with fungi from which it depends for survival and for oak colonization. Some of these fungi are ophiostomatoid species of the Raffaelea genus, known as ambrosia fungi associated with ambrosia beetles. Some Raffaelea species exhibit phytopathogenic activity causing wilting and/or death of trees. The objective of the present study is to identify the association between P. cylindrus and Raffaelea species in Portuguese cork oak stands showing symptoms of disease and decline. Materials and Methods: A total of 300 adult insects were collected as they emerged from cork oak logs, sampled from symptomatic trees. Axenic isolates of Raffaelea species were obtained from the beetles and their galleries in the trunks and identified based on morphological features and molecular analysis of the SSU and LSU rDNA regions. Results: Two Raffaelea species were identified, i.e., R. montetyi and a novel Raffaelea species closely related to R. canadensis. The novel species is morphologically and genetically characterized in this study, and erected as Raffaelea quercina M.L. Inácio, E. Sousa & F. Nóbrega, sp. nov. Raffaelea quercina constitutes a new phytopathogenic fungal species associated with P. cylindrus and cork oak trees exhibiting symptoms of vegetative decline. Conclusions: Raffaelea species appear to have a significant role in cork oak decline. Future research on the association between P. cylindrus and Raffaelea species, encompassing the trans-European and North African wide-range of cork oak stands, would further clarify the relationships between ambrosia beetles, associated fungi and cork oak decline, contributing to a better understanding of the phenomena and for strategies aiming to halt the continuous decline of the unique cork oak stands enclosed in the Mediterranean basin. [ABSTRACT FROM AUTHOR]

Morphological and molecular studies were conducted to characterize the specific identity of 36 isolates of potato cyst nematodes (PCNs) recovered from soil samples collected in several potato producing areas of Algeria. Morphometric data revealed that 44% of isolates contained Globodera pallida alone, 28% contained Globodera rostochiensis alone and 28% mixtures of the two species. Morphometric values of cysts and second-stage juveniles were generally distributed with slight differences in the expected ranges for both Globodera species. Inter- and intraspecific morphometric variability in nematode isolates was noted. Molecular analysis using conventional multiplex PCR with species-specific primers and TaqMan real-time PCR confirmed the morphological identification. In addition, the distribution of both potato cyst nematode species throughout various parts of the country was investigated. In the central areas, the isolates of G. pallida alone dominate, whereas isolates of G. rostochiensis alone are more frequent in the southern areas. In the eastern regions, mixed isolates are more representative. Most isolates examined in the western areas are mixtures of the two species or G. rostochiensis alone. Comparatively, G. pallida remains the most widely distributed species in its geographic range. This study confirms the presence of two PCN species, G. pallida and G. rostochiensis, in Algeria and provides additional information on their biogeographic distribution. [ABSTRACT FROM AUTHOR]

Potato is the third most important crop in the world after rice and wheat, with a great social and economic importance in Portugal as it is grown throughout the country, including the archipelagos of Madeira and the Azores. The tropical root-knot nematode (RKN) Meloidogyne luci is a polyphagous species with many of its host plants having economic importance and the ability to survive in temperate regions, which pose a risk to agricultural production. In 2019, M. luci was detected from soil samples collected from the council of Santo António in Pico Island (Azores). Bioassays were carried out to obtain females, egg masses, and second-stage juveniles to characterize this isolate morphologically, biochemically, and molecularly. The observed morphological features and morphometrics showed high similarity and consistency with previous descriptions. Concerning the biochemical characterization, the esterase (EST) phenotype displayed a pattern with three bands similar to the one previously described for M. luci and distinct from M. ethiopica. Regarding the molecular analysis, an 1800 bp region of the mitochondrial DNA between cytochrome oxidase subunit II (COII) and 16S rRNA genes was analyzed and the phylogenetic tree revealed that the isolate grouped with M. luci isolates (99.17%). This is the first report of M. luci parasitizing potato in the Azores islands, contributing additional information on the distribution of this plant-parasitic nematode. [ABSTRACT FROM AUTHOR]

Ambrosia beetles, including Platypus cylindrus, commonly act as vectors of Ophiostomatales. In an investigation of fungi associated with P. cylindrus, several species of Ophiostomatales were identified, including two species of Ophiostoma s. str. These two species were characterized based on comparisons of DNA sequences of the internal transcribed spacer regions including the 5.8S gene of rDNA and β‐tubulin gene. Two distinct clades were discernable in the phylogenetic trees, supported by high bootstrap values. One clade included the species Ophiostoma tsotsi, reported herein for the first time in Tunisia, and the species Ophiostoma quercus. [ABSTRACT FROM AUTHOR]