Ophiostomatoid fungi associated with declined Pinus pinaster stands in Spain

  • Gonzalo Álvarez Instituto Universitario de Investigación y Gestión Forestal Sostenible. Universidad de Valladolid-INIA. Palencia.
  • Mercedes Fernández Instituto Universitario de Investigación y Gestión Forestal Sostenible. Universidad de Valladolid-INIA. Palencia.
  • Julio J. Diez Instituto Universitario de Investigación y Gestión Forestal Sostenible. Universidad de Valladolid-INIA. Palencia.


Aim of study: We studied the presence of fungi and distribution patterns in relation to the health status of declining Pinus pinaster trees.

Area of study: Trees in two declining stands in Central Spain were allotted to three declining classes...

Material and Methods: Trees in two declining stands in Central Spain were allotted to three declining classes (healthy, declining and recently dead) and 3 trees of each class were felled in each stand. Wood slides (phloem and xylem) were taken at six positions along the trees and samples collected from fungal identification.

Main results: A total of 21 fungal taxa were isolated and identified; eleven of these species belonged to the Ophiostomatoid group. Ophiostoma minus was the most frequently isolated fungus and was identified in 22% of the samples, mainly associated to dead and diseased trees.

Research highlights: Together these results suggest a putative association of O. minor with the decline in this area, and thus we suggest paying more attention to this fungus as a potential agent of decline in P. pinaster stands.

Key words: Ophiostomatoid fungi; forest pathology; bluestain fungi; multivariate analyses.

Abbreviations: UTM: Universal Transverse Mercator coordinate system; MEA: Malt Extract Agar; DNA: Deoxyribonucleid acid; ITS: Internal Transcribed Spacer; BLAST: Basic Local Alignment Search Tool; CCA: Canonical Correspondence Analyses; PWN: Pine Wood Nematode.


Download data is not yet available.

Author Biography

Julio J. Diez, Instituto Universitario de Investigación y Gestión Forestal Sostenible. Universidad de Valladolid-INIA. Palencia.
Professor in Forest Pathology/Catedrático de Patología Forestal



Alvarez G, Fernández M, Diez JJ, 2008a. Hongos asociados al decaimiento del pino pinaster. Master Thesis, University of Valladolid. Spain.

Alvarez G, Fernández M, Diez JJ, 2008b. Hongos asociados al decaimiento de Pinus pinaster en España. Proc. XIV Congreso Sociedad Española de Fitopatología. Lugo (Spain). September 15-19.

Alvarez G, Fernández M, Diez JJ, 2009. Ophiostoma minus asociado a masas de Pinus pinaster (Aiton) con síntomas de decaimiento en Castilla y León. Proc. V Congreso Forestal Español. Ávila (Spain). September 21-25.

Barnett HL, Hunter BB, 1998. Illustrated genera of imperfect fungi. The American Phytopathological Society. St. Paul, Minnesota. USA. 218 pp.

Botella L, Diez, JJ, 2011. Phylogenic diversity of fungal enddophytes in Spanish stands of Pinus halepensis. Fungal Divers 47(1), 9–18. http://dx.doi.org/10.1007/s13225-010-0061-1

Botella L, Santamaría O, Díez JJ, 2010. Fungi associated with the decline of Pinus halepensis in Spain. Fungal Divers 40, 1–11. http://dx.doi.org/10.1007/s13225-010-0025-5

Bueno A, Diez JJ, Fernández MM, 2010. Ophiostomatoid fungi transported by Ips sexdentatus (Coleoptera; Scolytidae) in Pinus pinaster in NW Spain. Silva Fennica 44 (3), 387-397. http://dx.doi.org/10.14214/sf.137

Del Río M, Bravo F, Pando V, Sanz G, Sierra de Grado R, 2004. Influence of individual tree and stand attributes in stem straightness in Pinus pinaster Ait. stands. Ann For Sci 61, 141-148. http://dx.doi.org/10.1051/forest:2004005

DOE, 2009. Decreto 268/2009, de 28 de diciembre, por el que se adoptan medidas fitosanitarias para la erradicación y control del nematodo de la madera del pino Bursaphelenchus xylophilus (Steiner et Buhrer) Nickle et al., se regulan las indemnizaciones derivadas de la adopción de estas medidas y se dictan sus normas de aplicación para el año 2009 en la Comunidad Autónoma de Extremadura (Spain). Diario Oficial de Extremadura 248, pp 35397-35415.

DOG, 2012. Resolución do 16 de abril do 2012, da Secretaría Xeral de Medio Rural e Montes, polo que se establecen as restricións ás autorizacións de cortas de especies sensibles ao nematodo do piñeiro (Bursaphelenchus xylophilus) na zona demarcada das Neves. Diario Oficial de Galicia (Spain). 77. pp 14879-14880.

Fernández MM, García AE, Lieutier F, 2004. Effects of various densities of Ophiostoma ips inoculations on Pinus sylvestris in north-western Spain. For. Path 34(4), 213-223. http://dx.doi.org/10.1111/j.1439-0329.2004.00360.x

Ganley RJ, Newcombe G, 2006. Fungal endophytes in seeds and needles of Pinus monticola. Mycol Res 110, 318–327. http://dx.doi.org/10.1016/j.mycres.2005.10.005

Goidanich G, 1990. Manuale di patologia vegetale. Edizioni Agricole della Calderini, Bologna, Italy. 1283 pp.

Grylls B, Seifert KA, 1999. A synoptic key to species of Ophiostoma, Ceratocystis and Ceratocystiopsis. In: Ceratocystis and Ophiostoma. Taxonomy, Ecology and Pathogenicity (Wingfield MJ, Seifert KA, Webber JF. eds). pp 261-268.

Hanlin RT, 1998. Illustrated Genera of Ascomycetes. The American Phytopathological Society. St. Paul, Minnesota, USA. 164 pp.

Hausner G, Iranpour M, Kim JJ, Breuil C, Davis CN, Gibb EA, Reid J, Loewen PC, Hopkin AA, 2005. Fungi vectored by the introduced bark beetle Tomicus piniperda in Ontario, Canada, and comments on the taxonomy of Leptographium lundbergii, Leptographium terebrantis, Leptographium truncatum and Leptographium wingfieldii. Can J Bot 83, 1222. http://dx.doi.org/10.1139/b05-095

Hofstetter RW, Cronin JT, Klepzig KD, Moser JC, Ayres MP, 2006a. Antagonisms, mutualisms and commensalisms affect outbreak dynamics of the southern pine beetle. Oecologia 147, 679-691. http://dx.doi.org/10.1007/s00442-005-0312-0

Hofstetter RW, Klepzig KD, Moser JC, Ayres MP, 2006b. Seasonal dynamics of mites and fungi and their interaction with southern pine beetle. Environmental Entomology 35, 22-30. http://dx.doi.org/10.1603/0046-225X-35.1.22

Hu HL, Jeewon R, Zhou DQ, Zhou TX, Hyde HD, 2007. Phylogenetic diversity of endophytic Pestalotiopsis species in Pinus armandii and Ribes spp.: evidence from rDNA and β- tubulin gene phylogenies. Fungal Divers 24, 1–22.

Jacobs K, Wingfield MJ, 2001. Leptographium species: tree pathogens, insect asociates, and agents of blue-stain. American Phytopathological Society, St. Paul, Minnesota, USA. 207 pp.

Jankowiak, R, 2006. Fungi associated with Tomicus piniperda in Poland and assessment of their virulence using Scots pine seedlings. Ann Forest Sci 63, 801-808. http://dx.doi.org/10.1051/forest:2006063

Jankowiak R, Rossa R, Bilanski P, 2007. Contribution to pathogenicity of three blue-stain fungi associated with the pine sawyer beetle (Monochamus galloprovincialis) (Coleoptera: Cerambycidae) to Scots pine in Poland. Phytopathol Polon 46, 37-46.

Jankowiak R, Rossa R, 2007. Filamentous fungi associated with Monochamus galloprovincialis and Acanthocinus aedilis (Coleoptera: Cerambycidae) in Scots pine. Pol Bot J 52, 143-149.

Kiffer E, Morelet M, 1999. Les deuteromycetes, classification et clés d'identification generique. INRA. París. France. 306 pp.

Kuekam GN, Wingfield MJ, Roux J, 2013. Ceratocystis species, including two new taxa, from Eucalyptus trees in South Africa. Aus Plant Pathol 42(3), 283-311.

Langstrom B, Solheim H, Hellqvist C, Gref R, 1993. Effects of pruning young Scots pines on host vigour and susceptibility to Leptographium wingfieldii and Ophiostoma minus, two blue-stain fungi associated with Tomicus piniperda. Eur J Forest Pathol 23, 400-415. http://dx.doi.org/10.1111/j.1439-0329.1993.tb00820.x

Lanier L, Joly P, Bondoux P, Bellemère A, 1978. Mycologie et Patologie Forestières. Masson, Paris. 487 pp.

Legendre P, Legendre L, 1998. Numerical Ecology. Elsevier, Amsterdam, Holland. 852 pp.

Lieutier F, Yart A, Garcia J, Ham MC, Morelet M, Levieux J, 1989. Blue-stain fungi associated with two bark beetle pests of Scots pine (Pinus sylvestris): a preliminary study of fungal aggressiveness to the host tree. Ann Sci Forest 46, 210-216.

Lombardero MJ, Ayres MP, Hofstetter RW, Moser JC, Lepzig KD, 2003. Strong indirect interactions of Tarsonemus mites (Acarina: Tarsonemidae) and Dendroctonus frontalis (Coleoptera: Scolytidae). Oikos 102, 243-252. http://dx.doi.org/10.1034/j.1600-0706.2003.12599.x

López SR, Romón P, Iturrondobeitia JC, Goldazarena A, 2007. Los escolítidos del País Vasco. Guía práctica para su identificación y control. Servicio Central de Publicaciones del Gobierno Vasco. Vitoria-Gasteiz, Spain. 198 pp.

Maehara N, Hata K, Futai K, 2005. Effect of blue-stain fungi on the number of Bursaphelenchus xylophilus (Nematoda: Aphelenchoididae) carried by Monochamus alternatus (Coleoptera: Cerambycidae). Nematology 7, 161-167. http://dx.doi.org/10.1163/1568541054879557

Martínez P, Alves F, Diez, J, 2012. In vitro and in vivo interactions between Trichoderma viride and Fusarium circinatum. Silva Fennica 46(3), 303–316.

Masuya H, Kaneko S, Yamaoka Y, 2003. Comparative virulence of blue-stain fungi isolated from Japanese red pine. J. For. Res 8, 83-88. http://dx.doi.org/10.1007/s103100300009

Otrosina WJ, Bannwart D, Roncadori, RW, 1999. Root-infecting fungi associated with decline of longleaf pine in the southeastern United States. Plant and Soil 217, 145-150. http://dx.doi.org/10.1023/A:1004645115446

Piou D, Lieutier F, 1989. Symptomatological observations and possible roles of Ophiostoma minus and Tomicus piniperda in the decline of Scots pine in the forest of Orleans. Ann Sci Forest 46, 39-53. http://dx.doi.org/10.1051/forest:19890104

Popp MP, Johnson JD, Lesney MS, 1995. Characterization of the induced response of slash pine to inoculation with bark beetle vectored fungi. Tree Physiology 15, 619-623. http://dx.doi.org/10.1093/treephys/15.9.619

Prieto-Recio C, Romeralo C, Bezos D, Martin-Garcia J, Martinez-Alvarez, P, Botella L, Diez JJ, 2012. First report of Heterobasidion annosum on Pinus pinaster in Spain. Plant Dis. 96, 770. http://dx.doi.org/10.1094/PDIS-10-11-0890-PDN

Romón P, Zhou XD, Iturrondobeitia JC, Wingfield MJ, Goldazarena A, 2007. Ophiostoma species (Ascomycetes: Ophiostomatales) associated with bark beetles (Coleoptera: Scolytinae) colonizing Pinus radiata in northern Spain. Can J Microbiol 53, 756-767. http://dx.doi.org/10.1139/W07-001

Santamaria O, Diez JJ, 2005. Fungi in leaves, twigs and stem bark of Populus tremula from northern Spain. Forest Pathol 35, 95–104. http://dx.doi.org/10.1111/j.1439-0329.2004.00389.x

Scott JJ, Dong-Chan O, Yuceer C, Klepzig M, Clardy KD, Curie CR, 2008. Bacterial protection of beetle-fungus mutualism. Science 322, 63. http://dx.doi.org/10.1126/science.1160423

Serrada, R, 2004. Apuntes de Selvicultura. Fundación Conde del Valle Salazar. Escuela de Ingeniería Técnica Forestal. Madrid.

Six DL, Klepzig KD, 2004. Dendroctonus bark beetles as model systems for studies on symbiosis. Symbiosis 37, 207-232.

Solheim H, Langstrom B, Hellqvist C, 1993. Pathogenicity of the blue-stain fungi Leptographium wingfieldii and Ophiostoma minus to Scots pine: effect of tree pruning and inoculum density. Can. J. For. Res 23, 1438-1443. http://dx.doi.org/10.1139/x93-181

Solheim H, Langstrom B, 1991. Blue-stain fungi associated with Tomicus piniperda in Sweden and preliminary observations on their pathogenicity. Ann Sci Forest 48, 149-156. http://dx.doi.org/10.1051/forest:19910203

Sutton BC, 1980. The coelomycetes. Fungi imperfecti with pycnidia, acervuli and stromata. Commonwealth Mycological Institute, England, UK. 696 pp.

Ter Braak CJF, Šmilauer P, 2002. CANOCO Reference Manual and CanoDraw for Windows User's Guide: Software for Canonical Community Ordination (version 4.5). Microcomputer Power (Ithaca NY). USA.

Togashi K, 2004. A new method for loading Bursaphelenchus xylophilus (Nematoda: Aphelenchoididae) on adult Monochamus alternatus (Coleoptera: Cerambycidae). J Econ Entomol 97, 941-945. http://dx.doi.org/10.1093/jee/97.3.941

Warren JE, Edwards OR, Linit MJ, 1995. Influence of bluestain fungi on laboratory rearing of pinewood nematode infested beetles. Fund Appl Nematol 18.

Wingfield MJ, Seifert KA, Webber JF, 1993. Ceratocystis and Ophiostoma. Taxonomy, Ecology and Pathogenicity. American Phytopathological Press, St. Paul, Minnesota, U.S.A. 293 pp.

Wolfaardt JF, Wingfield MJ, Kendrick WB, 1992. Synoptic key and computer database for identification of species of Ceratocystis sensu lato. S. Afr. J. Bot. 58, 277-285.

Zamora P, Martínez-Ruiz C, Diez JJ. 2008. Fungi in needles and twigs of pine plantations from northern Spain. Fungal Divers 30, 171–184.

Zhou XD, De Beer ZW, Wingfield BD, Wingfield MJ, 2001. Ophiostomatoid fungi associated with three pine-infesting bark beetles in South Africa. Sydowia 53, 290-300.

Zhou XD, De Beer ZW, Wingfield BD, Wingfield MJ, 2002. Infection sequence and pathogenicity of Ophiostoma ips, Leptographium serpens and L. lundbergii to pines in South Africa. Fungal Diversity 10, 229-240.

How to Cite
ÁlvarezG., FernándezM., & DiezJ. J. (2015). Ophiostomatoid fungi associated with declined Pinus pinaster stands in Spain. Forest Systems, 24(1), e006. https://doi.org/10.5424/fs/2015241-05707
Research Articles