First Report of Bursaphelenchus xylophilus Causing Pine Wilt Disease on Pinus radiata in Spain
2015; American Phytopathological Society; Volume: 99; Issue: 10 Linguagem: Inglês
10.1094/pdis-03-15-0252-pdn
ISSN1943-7692
AutoresPaula F. Zamora, Vanessa Díaz Rodríguez, F. Renedo, A. Sanz, Juan Carlos Dominguez, G. Pérez-Escolar, J. S. da S. Miranda, Bustamante Álvarez, A. González-Casas, Eric Mayor, Montserrat Dueñas, Alicia B. Miravalles, Alfonso Navas, Lee Robertson, A. B. Martín,
Tópico(s)Parasite Biology and Host Interactions
ResumoHomePlant DiseaseVol. 99, No. 10First Report of Bursaphelenchus xylophilus Causing Pine Wilt Disease on Pinus radiata in Spain PreviousNext DISEASE NOTES OPENOpen Access licenseFirst Report of Bursaphelenchus xylophilus Causing Pine Wilt Disease on Pinus radiata in SpainP. Zamora, V. Rodríguez, F. Renedo, A. V. Sanz, J. C. Domínguez, G. Pérez-Escolar, J. Miranda, B. Álvarez, A. González-Casas, E. Mayor, M. Dueñas, A. Miravalles, A. Navas, L. Robertson, and A. B. MartínP. ZamoraSearch for more papers by this author, V. RodríguezSearch for more papers by this author, F. RenedoSearch for more papers by this author, A. V. SanzSearch for more papers by this author, J. C. DomínguezSearch for more papers by this author, G. Pérez-EscolarSearch for more papers by this author, J. MirandaSearch for more papers by this author, B. ÁlvarezSearch for more papers by this author, A. González-CasasSearch for more papers by this author, E. MayorSearch for more papers by this author, M. DueñasSearch for more papers by this author, A. MiravallesSearch for more papers by this author, A. NavasSearch for more papers by this author, L. RobertsonSearch for more papers by this author, and A. B. MartínSearch for more papers by this authorAffiliationsAuthors and Affiliations P. Zamora , Centro de Sanidad Forestal de Calabazanos, Consejería de Fomento y Medio Ambiente, Dirección General del Medio Natural, Junta de Castilla y León, Polígono Industrial de Villamuriel de Cerrato, s/n. 34190 Palencia, España V. Rodríguez F. Renedo , Dirección General del Medio Natural, Servicio de Defensa del Medio Natural. Consejería de Fomento y Medio Ambiente. Junta de Castilla y León. C/Rigoberto Cortejoso, 14. 47071 Valladolid. España A. V. Sanz J. C. Domínguez G. Pérez-Escolar J. Miranda B. Álvarez A. González-Casas E. Mayor M. Dueñas A. Miravalles , Centro de Sanidad Forestal de Calabazanos. Consejería de Fomento y Medio Ambiente. Dirección General del Medio Natural. Junta de Castilla y León. Polígono Industrial de Villamuriel de Cerrato, s/n. 34190 Palencia. España A. Navas L. Robertson , Laboratorio Nacional de Referencia Para la Identificación de Nematodos Fitopatógenos. Museo Nacional de Ciencias Naturales. CSIC. Departamento de Biodiversidad y Biología Evolutiva. C/José Gutiérrez Abascal 2. 28006 Madrid. España A. B. Martín , Centro de Sanidad Forestal de Calabazanos. Consejería de Fomento y Medio Ambiente. Dirección General del Medio Natural. Junta de Castilla y León. Polígono Industrial de Villamuriel de Cerrato, s/n. 34190 Palencia. España. Published Online:31 Jul 2015https://doi.org/10.1094/PDIS-03-15-0252-PDNAboutSectionsSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat The Pine Wood Nematode (PWN) Bursaphelenchus xylophilus (Steiner & Buhrer 1934) Nickle 1970, causal agent of pine wilt disease (PWD), was detected in the European territory for the first time in 1999 in Portugal, vectored by Monochamus galloprovincialis pine sawyer beetles. PWN has since been reported in three Spanish regions bordering Portugal: Extremadura; Galicia; and recently, Castilla y León. Pinus pinaster and P. nigra have been the only pine species infected by PWN in these countries so far. A 156-ha mixed coniferous plantation, including 67 ha of 40-year-old P. pinaster and P. radiata trees, located at Sancti-Spiritus (ETRS89: 30T, X 211795m, Y 4509866m) (Salamanca, Castilla y León, Spain), 30 km away from the Portuguese border, was surveyed for PWN presence. Ninety-four P. radiata trees showing crown dieback were sampled between December 2013 and January 2014. Wood samples were obtained at breast and crown height by drilling, and twigs were sampled by pole pruning. After 15 days incubation at 25°C, the samples were extracted with the Baermann funnel technique. Nematodes were morphologically identified as having subcylindrical female tail, prominent straight vulvar flap at 70 to 80% of body length from anterior end, tail tip with bursa in males, long and pointed rostrum in male spicula, <30 µm in length, and limbs with angular curvature (Anonymous 2013). Genetic identification was carried out at the Centro de Sanidad Forestal de Calabazanos (Castilla y León Autonomous Government), and the Spanish National Reference Laboratory for Plant Pathogen Nematodes (National Natural Science Museum, Madrid). B. xylophilus were placed in 10-µl nematode lysis buffer (1× PCR buffer, 60 mg/ml protease K) incubated at 60°C for 2 h and heated to 94°C for 15 min, then centrifuged. Five µl of the supernatant was used for PCR amplification. The primers J10-1, 5′-GGTGTCTAGTATAATATCAGAG-3′ and J10-2Rc, 5′-GTGAATTAGTGACGACGGAGTG-3′ (Castagnone et al. 2005), were used in 50-ml reactions with 2× Accuzyme (Bioline). Initial denaturation was carried out for a period of 2 min at 94°C followed by 35 cycles of 94°C for 1 min, 60°C for 1 min, 72°C for 1 min, and followed by a final 5-min extension period at 72°C. Sixty-nine sampled Monterey pines were found positive for PWN. In twenty-seven of these, presence of PWN in the different sampled tree parts was studied. Nematodes occurred up in the crown, either in the branches or in the upper trunk, but not in the base of the trunk in 15 trees that had live phloem. On the other hand, in 12 trees, nematodes had invaded the whole tree and the tree’s phloem was already dead. Although inoculation results had already shown that this species was susceptible to PWN (Dropkin et al. 1981; Evans et al. 1996, Furuno 1993), this is the first time that B. xylophilus is found naturally infecting trees of this species and causing PWD. Considering the high level of B. xylophilus natural infection in Monterey pines, this species should be regarded as susceptible to Pine Wilt Disease, at least when planted out of its natural range.References:Anonymous. 2013. EPPO Bull. 43:105. https://doi.org/10.1111/epp.12024 Crossref, Google ScholarCastagnone, C., et al. 2005. Eur. J. Plant Pathol. 112:191. https://doi.org/10.1007/s10658-004-0580-2 Crossref, ISI, Google ScholarDropkin, V. H., et al. 1981. Plant Dis. 65:1022. https://doi.org/10.1094/PD-65-1022 Crossref, ISI, Google ScholarEvans, H. F., et al. 1996. EPPO Bull. 26:199. Crossref, Google ScholarFuruno, T. 1993. Rep. Kyoto Univ. Forests. 25:20. 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