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3. Sampling and detection strategies for the pine pitch canker (PPC) disease pathogen Fusarium Circinatum in Europe

Author

European Commission, Ministerio de Economía y Competitividad (España), Fundação para a Ciência e a Tecnologia (Portugal), Vainio, Eeva [0000-0002-6739-7968], Bezos, Diana [0000-0002-0423-4627], Bragança, H. [0000-0002-4554-2722], Cleary, M. [0000-0002-0318-5974], Fourie, Gerda [0000-0003-2650-5448], Georgieva, M. [0000-0001-6580-4147], Ghelardini, L. [0000-0002-3180-4226], Hannunen, Salla [0000-0002-0998-4240], Ioos, Renaud [0000-0001-9359-5098], Martín-García, Jorge [0000-0002-3057-3710], Martínez-Álvarez, Pablo [0000-0002-4605-7702], Mullett, M. [0000-0002-6013-0347], Oszako, Tomasz [0000-0003-4688-2582], Papazova-Anakieva, Irena [0000-0002-7260-4685], Piškur, B. [0000-0002-9914-4930], Romeralo, Carmen [0000-0002-8510-9915], Sanz-Ros, Antonio V. [0000-0002-4791-8308], Steenkamp, Emma T. [0000-0003-0217-8219], Tubby, K. [0000-0001-6864-4185], Wingfield, Michael J. [0000-0001-9346-2009], Díez, Julio Javier [0000-0003-0558-8141], Vainio, Eeva, Bezos, Diana, Bragança, H., Cleary, M., Fourie, Gerda, Georgieva, M., Ghelardini, L., Hannunen, Salla, Ioos, Renaud, Martín-García, Jorge, Martínez-Álvarez, Pablo, Mullett, M., Oszako, Tomasz, Papazova-Anakieva, Irena, Piškur, B., Romeralo, Carmen, Sanz-Ros, Antonio V., Steenkamp, Emma T., Tubby, K., Wingfield, Michael J., Díez, Julio Javier, European Commission, Ministerio de Economía y Competitividad (España), Fundação para a Ciência e a Tecnologia (Portugal), Vainio, Eeva [0000-0002-6739-7968], Bezos, Diana [0000-0002-0423-4627], Bragança, H. [0000-0002-4554-2722], Cleary, M. [0000-0002-0318-5974], Fourie, Gerda [0000-0003-2650-5448], Georgieva, M. [0000-0001-6580-4147], Ghelardini, L. [0000-0002-3180-4226], Hannunen, Salla [0000-0002-0998-4240], Ioos, Renaud [0000-0001-9359-5098], Martín-García, Jorge [0000-0002-3057-3710], Martínez-Álvarez, Pablo [0000-0002-4605-7702], Mullett, M. [0000-0002-6013-0347], Oszako, Tomasz [0000-0003-4688-2582], Papazova-Anakieva, Irena [0000-0002-7260-4685], Piškur, B. [0000-0002-9914-4930], Romeralo, Carmen [0000-0002-8510-9915], Sanz-Ros, Antonio V. [0000-0002-4791-8308], Steenkamp, Emma T. [0000-0003-0217-8219], Tubby, K. [0000-0001-6864-4185], Wingfield, Michael J. [0000-0001-9346-2009], Díez, Julio Javier [0000-0003-0558-8141], Vainio, Eeva, Bezos, Diana, Bragança, H., Cleary, M., Fourie, Gerda, Georgieva, M., Ghelardini, L., Hannunen, Salla, Ioos, Renaud, Martín-García, Jorge, Martínez-Álvarez, Pablo, Mullett, M., Oszako, Tomasz, Papazova-Anakieva, Irena, Piškur, B., Romeralo, Carmen, Sanz-Ros, Antonio V., Steenkamp, Emma T., Tubby, K., Wingfield, Michael J., and Díez, Julio Javier

Abstract

Fusarium circinatum Nirenberg & O'Donnel is listed among the species recommended for regulation as quarantine pests in Europe. Over 60 Pinus species are susceptible to the pathogen and it also causes disease on Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) and species in genera such as Picea and Larix. The European Food Safety Authority considers the probability of new introductions-via contaminated seeds, wood material, soil and growing substrates, natural means and human activities-into the EU very likely. Due to early detection, constant surveillance and control measures, F. circinatum outbreaks have officially been eradicated in Italy and France. However, the global spread of F. circinatum suggests that the pathogen will continue to be encountered in new environments in the future. Therefore, continuous surveillance of reproductive material, nurseries and plantations, prompt control measures and realistic contingency plans will be important in Europe and elsewhere to limit disease spread and the "bridgehead effect", where new introductions of a tree pathogen become increasingly likely as new environments are invaded, must be considered. Therefore, survey programs already implemented to limit the spread in Europe and that could be helpful for other EU countries are summarized in this review. These surveys include not only countries where pitch canker is present, such as Portugal and Spain, but also several other EU countries where F. circinatum is not present. Sampling protocols for seeds, seedlings, twigs, branches, shoots, soil samples, spore traps and insects from different studies are collated and compiled in this review. Likewise, methodology for morphological and molecular identification is herein presented. These include conventional PCR with a target-specific region located in the intergenic spacer region, as well as several real-time PCR protocols, with different levels of specificity and sensitivity. Finally, the global situation and future perspec

Published
2019

4. Vertical transmission of Fusarium circinatum mitoviruses FcMV1 and FcMV2-2 via microconidia

Author

Ministerio de Economía y Competitividad (España), European Commission, Romeralo, Carmen [0000-0002-8510-9915], Bezos, Diana [0000-0002-0423-4627], Martínez-Álvarez, Pablo [0000-0002-4605-7702], Díez, Julio Javier [0000-0003-0558-8141], Romeralo, Carmen, Bezos, Diana, Martínez-Álvarez, Pablo, Díez, Julio Javier, Ministerio de Economía y Competitividad (España), European Commission, Romeralo, Carmen [0000-0002-8510-9915], Bezos, Diana [0000-0002-0423-4627], Martínez-Álvarez, Pablo [0000-0002-4605-7702], Díez, Julio Javier [0000-0003-0558-8141], Romeralo, Carmen, Bezos, Diana, Martínez-Álvarez, Pablo, and Díez, Julio Javier

Abstract

Pine Pitch Canker disease, caused by the pathogenic fungus Fusarium circinatum, affects conifer species worldwide. However, the virulence of the pathogen may be affected by the presence of mycoviruses. The aim of this laboratory-based study was to investigate the probability and rate of transmission of F. circinatum mitoviruses FcMV1 and FcMV2-2 via microconidia. Ten isolates of mitovirus-infected F. circinatum were subcultured to produce a total of 100 single-spore colonies (ten replicates per isolate). The total RNA and cDNA obtained from each spore isolate (monosporic culture) were amplified by PCR with specific primers for detection of F. circinatum mitoviruses FcMV1 and FcMV2-2. The mitoviruses were detected in a high percentage of the individual spore isolates (between 60% and 100% depending on the fungal isolate). However, the probability of transmission was not statistically significantly associated with either the F. circinatum isolate or the viral strain. A high proportion of transmission via microconidia is critical for development of a biological control program against Pine Pitch Canker (PPC) disease in forests. However, further studies are needed to establish the effect of these mitoviruses on the virulence of F. circinatum.

Published
2018

10. Sampling and Detection Strategies for the Pine Pitch Canker (PPC) Disease Pathogen Fusarium circinatum in Europe.

Author

Vainio, Eeva J., Bezos, Diana, Bragança, Helena, Cleary, Michelle, Fourie, Gerda, Georgieva, Margarita, Ghelardini, Luisa, Hannunen, Salla, Ioos, Renaud, Martín-García, Jorge, Martínez-Álvarez, Pablo, Mullett, Martin, Oszako, Tomasz, Papazova-Anakieva, Irena, Piškur, Barbara, Romeralo, Carmen, Sanz-Ros, Antonio V., Steenkamp, Emma T., Tubby, Katherine, and Wingfield, Michael J.

Subjects
FUSARIOSIS, INSECT trapping, DOUGLAS fir, PINE, SOIL sampling, PHYTOSANITATION, PINACEAE
Abstract

Fusarium circinatum Nirenberg & O'Donnel is listed among the species recommended for regulation as quarantine pests in Europe. Over 60 Pinus species are susceptible to the pathogen and it also causes disease on Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) and species in genera such as Picea and Larix. The European Food Safety Authority considers the probability of new introductions—via contaminated seeds, wood material, soil and growing substrates, natural means and human activities—into the EU very likely. Due to early detection, constant surveillance and control measures, F. circinatum outbreaks have officially been eradicated in Italy and France. However, the global spread of F. circinatum suggests that the pathogen will continue to be encountered in new environments in the future. Therefore, continuous surveillance of reproductive material, nurseries and plantations, prompt control measures and realistic contingency plans will be important in Europe and elsewhere to limit disease spread and the "bridgehead effect", where new introductions of a tree pathogen become increasingly likely as new environments are invaded, must be considered. Therefore, survey programs already implemented to limit the spread in Europe and that could be helpful for other EU countries are summarized in this review. These surveys include not only countries where pitch canker is present, such as Portugal and Spain, but also several other EU countries where F. circinatum is not present. Sampling protocols for seeds, seedlings, twigs, branches, shoots, soil samples, spore traps and insects from different studies are collated and compiled in this review. Likewise, methodology for morphological and molecular identification is herein presented. These include conventional PCR with a target-specific region located in the intergenic spacer region, as well as several real-time PCR protocols, with different levels of specificity and sensitivity. Finally, the global situation and future perspectives are addressed. [ABSTRACT FROM AUTHOR]

Published
2019
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