Joana M. S. Cardoso

University of Coimbra

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Isabel Abrantes

University of Coimbra

Luís Fonseca

University of Coimbra

Solange Oliveira

University of Évora

Maria do Rosário Félix

University of Évora

María Clara

University of Évora

Hugo Silva

University of Coimbra

Ivânia Esteves

University of Coimbra

Conceição Egas

University of Coimbra

Carla Maleita

University of Coimbra

Bruno Manadas

University of Coimbra

Cooperative Institutions

University of Coimbra

Consejo Superior de Investigaciones Científicas

University of Évora

Universidade Nova de Lisboa

University of Minho

Centre National de la Recherche Scientifique

Museo Nacional de Ciencias Naturales

Institute of Marine Research

Biocant

Instituto Nacional de Investigação Agrária e Veterinária

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Gene silencing in Bursaphelenchus xylophilus : knock down of a calponin gene and its effect on nematodes movement

Berichte aus dem Julius Kühn-Institut (2013)

Joana M. S. Cardoso Luís Fonseca Paula Gomes Conceição Egas Isabel Abrantes

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Complete genome sequence of a Tobacco necrosis virus D isolate from olive trees

Archives of Virology (2009)

Joana M. S. Cardoso Maria do Rosário Félix María Clara Solange Oliveira

Medical microbiology

Olive trees

Sequence (biology)

10.1007/s00705-009-0414-9

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Citations (13)

Bursaphelenchus tusciae in Tunisia associated with Hylurgus ligniperda

Forest Pathology (2016)

M. Mejri Luís Fonseca Joana M. S. Cardoso Mohamed Lahbib Ben Jamâa Isabel Abrantes

Summary Bursaphelenchus tusciae is reported for the first time in Tunisia and North Africa, associated with the insect Hylurgus ligniperda Fabricius (Coleoptera: Curculionidae: Scolytinae). Nematode identification was based on restriction fragment length polymorphism analysis and sequencing of the internal transcribed spacer (ITS) regions of ribosomal DNA. Phylogenetic analysis revealed that Tunisian B. tusciae clusters together with two other B. tusciae isolates forming a separate group close to B. hildegardae and B. eggersi . As H. ligniperda is among maritime pine scolytids pests in Tunisia and is widely distributed in North Africa, this study is an important contribution to the knowledge of Bursaphelenchus species associated with bark beetles of pine forests in Tunisia and North Africa.

10.1111/efp.12297

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Bursaphelenchus xylophilus in Pinus sylvestris—The First Report in Europe

Forests (2024)

Luís Fonseca Hugo Silva Joana M. S. Cardoso Ivânia Esteves Carla Maleita

The pinewood nematode (PWN), Bursaphelenchus xylophilus, is the causal agent of pine wilt disease and is considered an A2 quarantine organism by the European Plant Protection Organisation. In Europe, this nematode has been reported in Pinus pinaster, P. radiata, and P. nigra. In May 2024, severe wilting symptoms were observed in P. sylvestris trees at Serra da Lousã (Coimbra, the central area of continental Portugal). Wood samples were collected from six wilted trees, and the presence of PWN was investigated. From these, B. xylophilus specimens were detected in five out of the six trees. Species identification was performed based on species-specific morphological diagnostic characters, and this was confirmed by real-time PCR using species-specific primers targeting the B. xylophilus satellite DNA region. This study presents the first detection of B. xylophilus in P. sylvestris in Portugal and in Europe.

Xylophilus

10.3390/f15091556

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Species-Specific Molecular Detection of the Root Knot Nematode Meloidogyne luci

Biology (2021)

Carla Maleita Joana M. S. Cardoso Leidy Rusinque Ivânia Esteves Isabel Abrantes

Meloidogyne luci has been identified in various countries around the world parasitizing economically important crops and, due to its potential to cause serious damage to agriculture, was included in the European and Mediterranean Plant Protection Organization Alert List in 2017. This species shares morphological and molecular similarities with M. ethiopica and M. inornata, and a M. ethiopica group was therefore established. Although specific primers for the DNA amplification of species belonging to the M. ethiopica group have been developed previously, the primers were not species-specific, so molecular markers for the specific detection of M. luci are still needed. The objective of this study was to develop a SCAR marker for the detection of M. luci and the discrimination from other Meloidogyne spp. based on the intraspecific variability found in RAPD markers. RAPD screening of M. luci and M. ethiopica genome was used for the identification of a specific amplification product on M. luci, which was cloned, sequenced and converted into a SCAR marker. The specificity of the designed primers (Mlf/r) was tested and produced a fragment (771 bp) for all nine M. luci isolates with no amplification for the other nine Meloidogyne spp., including M. ethiopica and M. inornata. Additionally, the proper amplification of the M. luci SCAR-marker was also successful with DNA from galls of M. luci infected tomato roots. The results obtained in this study reveal that the specific molecular detection of M. luci was achieved and that the developed methodology can be used for routine diagnosis purposes, which are essential to monitoring the distribution and spread of M. luci in order to implement future effective and integrated nematode pest management programs.

Molecular marker

10.3390/biology10080775

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Meloidogyne luci, a new root‐knot nematode parasitizing potato in Portugal

Plant Pathology (2017)

Carla Maleita Ivânia Esteves Joana M. S. Cardoso Maria José da Cunha R. M. D. G. Carneiro

In 2013, during a field survey conducted in Portugal on potato, Solanum tuberosum , an unusual esterase ( EST ) phenotype was detected in a root‐knot nematode ( RKN ) from potato roots collected in Coimbra. This Portuguese isolate was purified and maintained on tomato, S. lycopersicum , and morphological, biochemical and molecular characteristics were studied. Perineal pattern morphology was highly variable, similar to Meloidogyne ethiopica and not useful for identification. The EST phenotype, from young egg‐laying females, displayed three bands similar to the Brazilian M. luci (L3) and distinct from M. ethiopica (E3). Phylogenetic analyses of mitochondrial cytochrome oxidase subunit I and the mitochondrial DNA region between COII and 16S rRNA genes revealed that the Portuguese isolate grouped with M. luci isolates close to M. ethiopica isolates. However, considering the ITS 1‐5.8S‐ ITS 2 region, the Portuguese isolate grouped with isolates of M. luci , M. ethiopica and M. hispanica , which limits the confidence of this region for M. luci diagnosis, and its differentiation from other species with morphological similarities. The M. luci pathogenicity to potato was also assessed in 16 commercial cultivars and compared with M. chitwoodi , considered to be a quarantine RKN species by EPPO . All potato cultivars were susceptible to both Meloidogyne species with gall indices of 5 and higher reproduction factor values ranging from 12.5 to 122.3, which suggests that M. luci may constitute a potential threat to potato production. In the present study, M. luci is reported for the first time attacking potato in Portugal.

Root-knot nematode

10.1111/ppa.12755

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Citations (38)

Direct molecular detection of the pinewood nematode, Bursaphelenchus xylophilus, from pine wood, bark and insect vector

European Journal of Plant Pathology (2011)

Joana M. S. Cardoso Luís Fonseca Isabel Abrantes

The pinewood nematode (PWN), Bursaphelenchus xylophilus, is the causal agent of pine wilt disease. The international economic impact of the introduction of the PWN into new areas has highlighted the need for the development of accurate and reliable detection methods of B. xylophilus, which are essential to define aspects of its control and management. In the present study, a methodology was developed for the direct detection of PWN by conventional PCR assay, with a species specific set of primers based on PWN satellite DNA, using total DNA extracted directly from maritime pine, Pinus pinaster, wood and bark samples, and from the insect vector, Monochamus galloprovincialis. This methodology involves homogenisation of wood, bark and insects using liquid nitrogen, DNA extraction and one or two PCR amplification steps, which permit the rapid and direct detection of one single nematode present in 100 mg of wood and bark and in one entire insect without the preliminary steps of nematode extraction.

Pinus pinaster

Xylophilus

Wilt disease

Pine wood

10.1007/s10658-011-9915-y

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Citations (32)

Virulence Biomarkers of Bursaphelenchus xylophilus: A Proteomic Approach

Frontiers in Plant Science (2022)

Joana M. S. Cardoso Sandra I. Anjo Bruno Manadas Hugo Silva Isabel Abrantes

The pinewood nematode (PWN), Bursaphelenchus xylophilus, one of the most serious forest pests worldwide, is considered the causal agent of the pine wilt disease (PWD). The main host species belong to the genus Pinus, and a variation in the susceptibility of several pine species to PWN infection is well-known. It is also recognized that there is variation in the virulence among B. xylophilus isolates. In the present study, we applied a quantitative mass spectrometry-based proteomics approach to perform a deep characterization of proteomic changes across two B. xylophilus isolates with different virulence from different hosts and geographical origins. A total of 1,456 proteins were quantified and compared in the two isolates secretomes, and a total of 2,741 proteins were quantified and compared in the nematode proteomes in pine tree extract and fungus stimuli conditions. From the proteomic analyses, a group of proteins was selected and identified as potential virulence biomarkers and shed light on putative most pathogenic proteins of this plant-parasitic nematode. Proteomic data are available via ProteomeXchange with identifier PXD029377.

Xylophilus

Wilt disease

Proteome

Pinus thunbergii

10.3389/fpls.2021.822289

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Citations (10)

Bursaphelenchus fungivorus from Pinus pinaster bark in Portugal

Forest Pathology (2013)

Luís Fonseca Joana M. S. Cardoso Jesús Morón-López Isabel Abrantes

Summary Bursaphelenchus fungivorus is reported for the first time in P ortugal, identified as associated with P inus pinaster bark and characterized on the basis of morphological and morphometrical characters for this species. Species identification was confirmed using restriction fragment length polymorphism analysis and sequencing of the internal transcribed spacer ( ITS ) regions of ribosomal DNA . Intraisolate genetic variability was detected among ITS sequences of the P ortuguese B . fungivorus isolate. Phylogenetic analysis, obtained from multiple sequence alignment between ITS sequences of B ursaphelenchus species, revealed that the P ortuguese B . fungivorus isolate clusters with other B . fungivorus isolates, forming a separate group close to B . seani highlighting a molecular proximity of these two species.

Spacer DNA

10.1111/efp.12077

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Citations (5)

Pine wilt disease: what do we know from proteomics?

BMC Plant Biology (2024)

Joana M. S. Cardoso Bruno Manadas Isabel Abrantes Lee Robertson Susana C. Arcos

Abstract Pine wilt disease (PWD) is a devastating forest disease caused by the pinewood nematode (PWN), Bursaphelenchus xylophilus , a migratory endoparasite that infects several coniferous species. During the last 20 years, advances have been made for understanding the molecular bases of PWN-host trees interactions. Major advances emerged from transcriptomic and genomic studies, which revealed some unique features related to PWN pathogenicity and constituted fundamental data that allowed the development of postgenomic studies. Here we review the proteomic approaches that were applied to study PWD and integrated the current knowledge on the molecular basis of the PWN pathogenicity. Proteomics has been useful for understanding cellular activities and protein functions involved in PWN-host trees interactions, shedding light into the mechanisms associated with PWN pathogenicity and being promising tools to better clarify host trees PWN resistance/susceptibility.

Wilt disease

Host resistance

10.1186/s12870-024-04771-9

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Citations (8)