Paloma Abad‐Campos

Universitat Politècnica de València

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Co-Authors

Josep Armengol

Universitat Politècnica de València

A. Pérez‐Sierra

Instituto Valenciano de Investigaciones Agrarias

Maela León

Universitat Politècnica de València

J. García‐Jiménez

Benemérita Universidad Autónoma de Puebla

José Vicente García-Jiménez

Universidad de Murcia

Antonio Vicent Civera

Instituto Valenciano de Investigaciones Agrarias

Mónica Berbegal

Universitat Politècnica de València

Sandra Alaniz

Universidad de la República

L. A. Álvarez

Universidad Nacional Agraria La Molina

David Gramaje

Consejo Superior de Investigaciones Científicas

Cooperative Institutions

Universitat Politècnica de València

Consejo Superior de Investigaciones Científicas

Universitat de València

United States Department of Agriculture

Agricultural Research Service

Mendel University in Brno

Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement

University of Algarve

Forest Research

University of Sousse

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Outbreak of Pitch Canker Caused by Fusarium circinatum on Pinus spp. in Northern Spain

Plant Disease (2005)

E. Landeras P. García Yaisely Orquidea Hernandez Fernandez M. Braña O. Fernández-Alonso

During the winter of 2003-2004, dieback symptoms were observed on Pinus radiata and P. pinaster in pine nurseries in Asturias (northern Spain). Small groups of affected seedlings appeared randomly distributed throughout the nurseries. The seedlings died rapidly, showing basal needle dieback, stem lesions, resin exudations, and wilting. Isolations from infected material onto potato dextrose agar (PDA) supplemented with 0.5 mg/ml of streptomycin sulfate and Komada's medium consistently yielded Fusarium sp. cultures. The isolates were transferred to PDA and Spezieller Nährstoffarmer agar and incubated at 25°C for 10 days with a 12-h photoperiod. The cultures were identified as Fusarium circinatum Nirenberg & O'Donnell (= Fusarium subglutinans Wollenweb. & Reinking), causal agent of pitch canker disease, on basis of the presence of polyphialides and characteristic sterile, coiled, hyphae (2). To further confirm their identity, a polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) based on histone H3 gene sequences (4) and a test based on the F. circinatum-specific primers, CIRC1A-CIRC4A, which amplifies a 360-bp DNA fragment of the intergenic spacer region of the nuclear ribosomal operon (3), were used. Results obtained with both techniques confirmed the morphological identification of the cultures. A representative culture has been placed in the Centraalbureau voor Schimmelcultures (CBS 117843). The pathogen was isolated only from seedlings of P. radiata and P. pinaster. Other species such as P. nigra, P. sylvestris, and Pseudotsuga menziesii, which were also grown in these nurseries, did not show symptoms. Pathogenicity was confirmed by inoculating 6- to 9-month-old P. radiata and P. pinaster seedlings. Small strips of bark (10 × 1 mm) were cut from the stems and similar sized pieces of PDA colonized by F. circinatum were placed in contact with the open wounds and covered with parafilm. Basal needle dieback was observed 10 days after inoculation that resulted in wilting of the seedlings. F. circinatum was reisolated from the affected stems fulfilling Koch's postulates. Later in the year, symptoms of pitch canker were also observed on 20-year-old P. radiata in one forest plantation in Cantabria (northern Spain). Infected branches and shoots of the trees exudated abundant resin, resulting in resinous cankers. The needles, distal to branch tip infections, wilt, fade to yellow then red, and fall from the tree. Affected trees showed noticeable crown dieback. The isolations from the cankers also yielded F. circinatum cultures that were identified as described above. Although a nonrefereed report appeared in 1998 (1), to our knowledge, this is the first report of F. circinatum on P. radiata and P. pinaster in Spain and in Europe. References: (1) L. D. Dwinell et al. Int. Congr. Plant Pathol. 7th. 3:9, 1998. (2) H. I. Nirenberg and K. O'Donnell. Mycologia 90:434, 1998. (3) W. Schweigkofler et al. Appl. Environ. Microbiol. 70:3512, 2004. (4) E. T. Steenkamp et al. Appl. Environ. Microbiol. 65:3401, 1999.

Pinus radiata

Canker

Potato dextrose agar

Collar rot

Intergenic region

10.1094/pd-89-1015a

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Citations (114)

Survey of Oomycetes Associated with Root and Crown Rot of Almond in Spain and Pathogenicity of Phytophthora niederhauserii and Phytopythium vexans to ‘Garnem’ Rootstock

Agriculture (2022)

Francisco Beluzán Xavier Miarnau Laura Torguet Josep Armengol Paloma Abad‐Campos

From 2018 to 2020, surveys of oomycetes associated with root and crown rot of almond (Prunus dulcis) were conducted on diseased young almond trees in commercial orchards and nurseries in six provinces of Spain. A total of 104 oomycete isolates were obtained from plant and soil samples, which h were identified by sequencing the internal transcribed spacer (ITS) region of the ribosomal DNA. Diverse species belonging to the genera Globisporangium, Phytophthora, Phytopythium and Pythium were found, Phytopythium vexans and Phytophthora niederhauserii being the most frequent. The pathogenicity of these two species to one-year-old almond seedlings of ‘Garnem’ (P. dulcis × P. persica) rootstock was studied. All seedlings inoculated with Pp. vexans and Ph. niederhauserii isolates showed severe symptoms at the late stage of the pathogenicity test (defoliation, wilting and dieback) and several plants died. Some isolates of Ph. niederhauserii significantly reduced the dry weight of the roots compared with the control, but this effect was not observed in seedlings inoculated with Pp. vexans. These results provide new information about the oomycete species present in almond crops in Spain and highlight the importance of carrying out frequent phytosanitary surveys for a better knowledge of potential risks posed by these soil-borne pathogens.

Oomycete

Root rot

Wilting

Prunus dulcis

Quercus robur

10.3390/agriculture12020294

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Citations (15)

The Use of Genus-Specific Amplicon Pyrosequencing to Assess Phytophthora Species Diversity Using eDNA from Soil and Water in Northern Spain

PLoS ONE (2015)

Santiago Català A. Pérez‐Sierra Paloma Abad‐Campos

Phytophthora is one of the most important and aggressive plant pathogenic genera in agriculture and forestry. Early detection and identification of its pathways of infection and spread are of high importance to minimize the threat they pose to natural ecosystems. eDNA was extracted from soil and water from forests and plantations in the north of Spain. Phytophthora-specific primers were adapted for use in high-throughput Sequencing (HTS). Primers were tested in a control reaction containing eight Phytophthora species and applied to water and soil eDNA samples from northern Spain. Different score coverage threshold values were tested for optimal Phytophthora species separation in a custom-curated database and in the control reaction. Clustering at 99% was the optimal criteria to separate most of the Phytophthora species. Multiple Molecular Operational Taxonomic Units (MOTUs) corresponding to 36 distinct Phytophthora species were amplified in the environmental samples. Pyrosequencing of amplicons from soil samples revealed low Phytophthora diversity (13 species) in comparison with the 35 species detected in water samples. Thirteen of the MOTUs detected in rivers and streams showed no close match to sequences in international sequence databases, revealing that eDNA pyrosequencing is a useful strategy to assess Phytophthora species diversity in natural ecosystems.

Pyrosequencing

Amplicon

Environmental DNA

10.1371/journal.pone.0119311

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Citations (92)

First Report of Leaf Spot and Twig Blight of Rhododendron spp. Caused by Phytophthora hibernalis in Spain

Plant Disease (2007)

L. A. Álvarez A. Pérez‐Sierra J. García‐Jiménez Paloma Abad‐Campos E. Landeras

During the early spring of 2004, an estimated 20% of containerized nursery stocks of Rhododendron spp. in Asturias (northern Spain) were affected by a foliar disease that has reoccurred annually. Leaf spots were dark brown to almost black, generally oval to round, visible from both sides of the leaf, and expanded to affect the entire leaf including the petiole. Affected leaves abscised from the plant. A Phytophthora sp. was consistently isolated from symptomatic leaf tissues on PARBH medium (3) and hyphal tips were transferred onto potato dextrose agar (PDA). Colonies grown on PDA at 20°C were submerged, had a growth rate of 2.2 mm/day, and had lobes of compact mycelium. Sporangia were semipapillate and caducous with a pedicel (20.0-) 37.7 (-52.5) μm long. Sporangia were asymmetrical in shape with the broadest point near the apex: 25.2 to 40.4 μm long × 10.2 to 15.8 μm wide (average 33.1 × 12.6 μm), and length/width ratio was 2.8:1. Chlamydospores were not observed. Isolates were homothallic and oogonia ranged from 26.5 to 27.5 μm in diameter. Antheridia were mostly amphigynous but occasionally paragynous. Oospores were plerotic and 23.1 to 25.5 μm in diameter. These characteristics conformed to those of Phytophthora hibernalis Carne (2). Sequences of the internal transcribed spacer regions on the isolates and comparison with other sequences in GenBank showed that they were identical to P. hibernalis (Accession No. AY827556.1 from Citrus sp.). For pathogenicity tests, four isolates of P. hibernalis were used to inoculate detached leaves of Rhododendron hybrid Brigitte. The underside of five detached leaves was inoculated with a drop of 40 μL of a suspension of 104 zoospores/ml. Controls were inoculated with a 40-μL drop of sterile distilled water. Leaves were incubated in a moist chamber at 20°C in the dark. A quantification of the lesion area was made 8 days after inoculation using the software Assess-APS. All inoculated leaves developed necrotic lesions that ranged from 0.246 to 1.512 cm2. P. hibernalis was reisolated from infected tissue. Symptoms were not detected on the controls. The test was repeated twice and similar results were obtained each time. P. hibernalis has been described previously as causing brown rot on citrus in Spain (4) and was isolated from rhododendron plants in California and Oregon (1). To our knowledge, this is the first record of P. hibernalis causing foliar blight on Rhododendron species in Spain as well as in Europe. References: (1) C. Blomquist et al. Online publication. doi:10.1094/PHP-2005-0728- 01-HN. Plant Health Progress, 2005. (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul MN. 1996. (3) S. N. Jeffers and S. B. Martin. Plant Dis. 70:1038, 1986. (4) J. J. Tuset. An. Inst. Nac. Investig. Agrar. Ser. Prot. Veg. N.7, 1977.

Pedicel

Petiole (insect anatomy)

Potato dextrose agar

Spots

Pycnidium

Antheridium

Chlamydospore

Oospore

Wilting

Apex (geometry)

10.1094/pdis-91-7-0909a

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Citations (7)

Novedades en la formulación de plaguicidas

Cuadernos de fitopatología: Revista técnica de fitopatología y entomología (1992)

A. Carreres Ortega Margarita Juárez Sanz Paloma Abad‐Campos

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First Report of Diaporthe amygdali Associated with Twig Canker and Shoot Blight of Nectarine in Spain

Plant Disease (2021)

Francisco Beluzán Diego Olmo Maela León Paloma Abad‐Campos Josep Armengol

Nectarine (Prunus persica (L.) Batsch var. nucipersica (Suckow) C. K. Schneid.) is a fruit crop widely cultivated throughout the Mediterranean basin. In Spain, it is mainly grown in eastern regions of the country. In March 2018, 5-year-old nectarine trees showing twig canker symptoms were observed after a rainy spring period in a 0.5 ha orchard located at Alaior, Menorca island (Spain). Cankers were frequent on affected trees (approximately, 80% of the total trees), thus leading to shoot blight. Ten twig segments of one-year old wood with cankers were cut, washed under running tap water, surface disinfected for 1 min in a 1.5% sodium hypochlorite solution and rinsed twice in sterile distilled water. Small pieces (2 mm) of affected tissues were taken from the margin of the cankers and plated on potato dextrose agar (PDA) supplemented with 0.5 g/L of streptomycin sulphate (PDAS). The plates were then incubated at 25 ºC in the dark for 7 to 10 d. Actively growing colonies were first hyphal-tipped and then transferred to PDA and 2% water agar supplemented with sterile pine needles and incubated at 21-22ºC under a 12h/12h near UV / darkness cycle during 21 d (León et al. 2020). Colonies were white at first, becoming light cream, with visible solitary and aggregate pycnidia at maturity. Alpha conidia were aseptate, fusiform, hyaline, multi-guttulated (mean ± SD = 7.4 ± 0.7 × 2.8 ± 0.4 µm, n = 100). Beta and gamma conidia were not observed. The morphological and cultural characteristics of the isolates were congruent with those of Diaporthe spp. (Gomes et al. 2013). The ITS1-5.8S-ITS2 (ITS) region and fragments of β-tubulin (tub2), the translation elongation factor 1-alpha (tef1-α) gene regions, histone H3 (his3) and calmodulin (cal) genes of representative isolate DAL-59 were amplified and sequenced (Santos et al. 2017). The BLASTn analysis revealed 100% similarity with sequences of D. mediterranea (Synonym D. amygdali) (Hilário et al. 2021) isolate DAL-34 from almond (ITS: MT007489, tub2: MT006686, tef1-α: MT006989, his3: MT007095 and cal: MT006761). Sequences of isolate DAL-59 were deposited in GenBank Database (ITS: MT007491, tub2: MT006688, tef1-α: MT006991, his3: MT007097 and cal: MT006763). Pathogenicity tests were conducted using one-year-old potted plants of nectarine cv. Boreal, which were inoculated with isolate DAL-59. In each plant, a 3 mm wound was made in the center of the main branch (about 30 cm length) with a scalpel. Colonized agar plugs with 3 mm diameter, which were obtained from active 10-day-old colonies growing on PDA, were inserted underneath the epidermis and the wounds sealed with Parafilm. Inoculated plants were incubated in a growth chamber at 23 ºC with 12 h of light per day. Controls were inoculated with uncolonized PDA plugs. There were twelve plants per treatment, which were arranged in a completely randomized design. Five days after inoculation necrosis development was observed in the area of inoculation. Wilting and twig blight symptoms over the lesion occurred 3-wk after inoculation and pycnidia were detected, while the controls remained asymptomatic. Diaporthe amygdali was re-isolated from symptomatic tissues and identified as described above to satisfy Koch's postulates. To our knowledge, this is the first report of D. amygdali causing twig canker and shoot blight disease on nectarine in Spain.

Canker

10.1094/pdis-10-20-2283-pdn

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Citations (2)

Patógenos fúngicos de cuarentena que afectan a los frutos cítricos. I: descripción

Levante Agrícola: Revista internacional de cítricos (2004)

Antonio Vicent Civera Paloma Abad‐Campos Josep Armengol José Vicente García-Jiménez

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Monosporascus eutypoides, a Cause of Root Rot and Vine Decline in Tunisia, and Evidence that M. cannonballus and M. eutypoides Are Distinct Species

Plant Disease (2012)

I. Ben Salem Kamila C. Correia N. Boughalleb Sami Jorge Michereff Maela León

Three Monosporascus eutypoides-like isolates recovered from cucurbit plants with symptoms of Monosporascus root rot and vine decline in Tunisia were compared to 28 isolates of M. cannonballus from 12 countries for phenotypic, genomic, and pathogenicity characteristics. Morphologically, M. cannonballus and M. eutypoides-like cultures were similar, each producing fertile perithecia in culture containing globose, smooth, dark brown to black ascospores. Nevertheless, all M. cannonballus isolates had one ascospore per ascus, while M. eutypoides-like isolates had mainly two to three ascospores per ascus (rarely one). The employment of the internal transcribed spacer (ITS) of nuclear ribosomal DNA, the elongation factor 1-α (EF-1α), and the β-tubulin (β-tub) gene sequence diversity analyses and the resulting phylogenies identified a level of polymorphism that enabled separation of M. cannonballus and M. eutypoides-like isolates. All isolates of M. cannonballus had identical EF-1α and β-tub sequences irrespective of very diverse geographic origins, which were different from the EF-1α and β-tub sequences of the M. eutypoides-like isolates (96 and 97% similarity, respectively). Similar results were obtained for the ITS region of rDNA. In addition, of three M. eutypoides-like isolates tested for pathogenicity, all three were pathogenic on watermelon, two were pathogenic on muskmelon, but only one was pathogenic on cucumber. The results demonstrate that the M. eutypoides-like isolates belong to the species M. eutypoides, and that M. cannonballus and M. eutypoides are distinct species.

Root rot

Ascus (bryozoa)

10.1094/pdis-05-12-0464-re

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Citations (23)

Phytophthora in forests and natural ecosystems

Paloma Abad‐Campos Ana Cristina Coelho Pilar Fernández‐Rebollo Luis V. García T. Jung

Natural forest

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Citations (52)

Detection of stolbur phytoplasma in willow in Spain

Bulletin of insectology (2011)

A. Alfaro‐Fernández Paloma Abad‐Campos D. Hernández-Llópis A. Serranofernández I. Font

Preliminary results of nested-PCR indicated that phytoplasmas were detected in willow (Salix babylonica Linn) showing yellows, ball-like structures and small leaves symptoms collected in Valencia Province (Eastern Spain). RFLP analyses showed that the phytoplasmas belonged to the stolbur group (16SrXII).

Phytoplasma

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Citations (5)