Fusarium oxysporum and F. verticillioides associated with damping-off in Pinus spp.

Caciara Gonzatto Maciel, Clair Walker, Ricardo Feliciano dos Santos, Marlove Fátima Brião Muniz, Daniele Lemos Brum

Resumo


Occurrence of Fusarium spp. is one of the problems, most limiting to growth of seedlings, in nurseries. This pathogen can be transmitted via seeds and causes damages to the seedlings during pre- and post-emergence stages. The present study aimed to identify Fusarium spp. at the species level based on morphological and molecular characteristics and to verify the pathogenicity of these isolates in seeds lots of Pinus elliottii and P. taeda. For this, we used two Fusarium isolates and five lots of Pinus spp. seeds. Morphological characterization was performed based on a key, specific to Fusarium spp. identification, whereas, molecular identification was carried out by amplification and sequencing of the regions from internal transcribed spacer (ITS) and the elongation factor 1-α (tef1). The pathogenicity test was conducted through the contact of the seeds with fungal culture for 48 h, followed by sowing them in sand. The variables evaluated were emergency speed index, percentage of emergency, non-emergency seeds, symptomatic seedlings, and seedling damping-off. One isolate, F1UFSM, was identified as F. verticillioides and another isolate, F2UFSM, was identified as F. oxysporum. Both the isolates were pathogenic to the seeds of Pinus spp., causing a reduction in the percentage of emergence and seedling damping-off.

Palavras-chave


ITS; Elongation factor (tef1); Pinus taeda; Pinus elliottii

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Referências


ALFENAS, A. C.; MAFIA, R. G. Métodos em fitopatologia. Viçosa: UFV, 2007. 382 p. BRASIL. Ministério da Agricultura, Pecuária e Abastecimento. Regra para análise de sementes. Brasília, DF, 2009. 395 p.

ARIF, M.; CHAWLA, S.; ZAIDI, N. W. et al. Development of specific primers for genus Fusarium and F. solani using rDNA sub-unit and transcription elongation factor (TEF-1α) gene. African Journal of Biotechnology, v. 11, n. 2, p. 444-447, 2012.

DELLAPORTA, S. L.; WOOD, J.; HICKS, J. B. A plant DNA minipreparation: version II. Plant Molecular Biology Reporter, v. 1, n. 4, p. 19-21, 1983.

FERREIRA, D. F. Sisvar: sistemas de análises de variância para dados balanceados: programa de análises estatísticas e planejamento de experimentos. versão 5.3 (Biud 75). Lavras: Universidade Federal de Lavras, 2010.

GEISER, D. M. et al. FUSARIUM-ID v.1.0: a DNA sequence database for identifying Fusarium. European Journal Plant Pathology, v. 110, n. 5, p. 473-479, 2004.

GERLACH, W.; NIRENBERG, H. The genus Fusarium: - a pictorial atlas. Berlin: Biologische Bundesanstaltfür Land - und. Forstwirtschoft, 1982. 406 p.

GRIGOLETTI JUNIOR, A.; AUER, C. G. Fusariose em mudas de Pinus taeda. Colombo: Embrapa Florestas, 2006. 3 p. (Comunicado técnico 166).

GUPTA, V. K.; MISRA, A. K.; GAUR, R. K. Growth characteristics of Fusarium spp. causing wilt disease in Psidium guajava l. in Índia. Journal of Plant Protection Research, v. 50, n. 4, p. 451-462, 2010.

LANDERAS, E. et al. Outbreak of pitch canker caused by Fusarium circinatum on Pinus spp. in Northern Spain. Plant Disease, v. 89, n. 9, p. 1015, 2005.

LAZAROTTO, M. et al. Mycelia growth and sporulation of Fusarium chlamydosporum species complex under different culture conditions. Revista de Ciências Agrárias, v. 57, n. 1, p. 35-40, 2014.

LESLIE, J. F.; SUMMERELL, B. A. The Fusarium laboratory manual. 1. ed. USA: Blackwell Publishing, 2006. 388 p.

LORI, G. A.; SALERNO, M. I. Fusarium species on seeds of Pinus taeda L. and Pinus elliottii Engelm. in Argentina. Journal of Plant Diseases and Protection, v. 110, n. 5, p. 437-443, 2003.

MACIEL, C. G. et al. First report of Fusarium sambucinum associated on Pinus elliottii seeds in Brazil. Plant Disease, v. 97, n. 7, p. 995, 2013.

MAGUIRE, J. D. Speed of germination aid in selection and evaluation for seeding emergence and vigor. Crop Science, v. 2, n. 2, p. 76-177, 1962.

MARTÍN-PINTO, P.; PAJARES, J.; DÍEZ, J. Pathogenicity of Fusarium verticillioides and Fusarium oxysporum on Pinus nigra seedlings in northwest Spain. Forest Pathology, v. 38, n. 2, p. 78-82, 2008.

MENEZES, J. P. et al. Variabilidade genética na região its do rDNA de isolados de Trichoderma spp. (Biocontrolador) e Fusarium oxysporum f. sp. chrysanthemi. Ciência e Agrotecnologia, v. 34, n. 1, p. 132-139, 2010.

NELSON, P. E.; TOUSSON, T. A.; MARASAS, W. F. O. Fusarium species: an illustrated manual for identification. Philadelphia: Pennsylvania State University Press, 1983. 193 p.

O’DONNELL, K. Molecular phylogeny of the Nectria haematococca-Fusarium solani species complex. Mycologia, v. 92, n. 5, p. 919-938, 2000.

OCAMB, C. M.; JUZWIK, J.; MARTIN, F. B. Fusarium spp. and Pinus strobus seedlings: root disease pathogens and taxa associated with seed. New Forests, v. 24, n. 1, p. 67-79, 2002.

PFENNING, L. H. et al. First report and characterization of Fusarium circinatum, the causal agent of pitch canker in Brazil. Tropical Plant Pathology, v. 39, n. 3, p. 210-216, 2014.

REHNER, S. A.; BUCKLEY, E. A Beauveria phylogeny inferred from nuclear ITS and EF1-a sequences: evidence for cryptic diversification and links to Cordyceps teleomorphs. Mycologia, v. 97, n.1, p. 84-98, 2005.

SCHMITZ, A.; RIESNER, D. Purification of nucleic acids by selective precipitation with polyethylene glycol 6000. Analytical Biochemistry, v. 354, n. 2, p. 311-313, 2006.

SCHOCH, C. L. et al. Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi. Proceedings of the National Academy of Sciences of the United States of America, v. 109, n. 16, p. 1-6, 2012.

SILVA, J. L.; TEIXEIRA, R. N. V. Esporulação e crescimento micelial de Fusarium solani em diferentes meios de cultura e regimes de luminosidade. Agro@mbiente On-line, v. 6, n. 1, p. 47-52, 2012.

SKOVGAARD, K. et al. Fusarium commune is a new species identified by morphological and molecular phylogenetic data. Mycologia, v. 95, n. 4, p. 630-636, 2003.

STEENKAMP, E. T. et al. Fusarium circinatum and pitch canker of Pinus in Colombia. Australasian Plant Pathology, v. 41, n. 5, p. 483-491, 2012.

TAMURA, K.; DUDLEY, J.; NEI, M. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biology and Evolution, v. 24, n. 8, p. 1596-1599, 2007.

TEIXEIRA, H.; VIEIRA, M.; MACHADO, J. C. Marcadores RAPD na análise da diversidade genética de isolados de Acremonium strictum. Fitopatologia Brasileira, v. 29, n. 6, p. 651-655, 2004.

TSUI, C. K. M.; WOODHALL, J.; CHEN, W. Molecular techniques for pathogen identification and fungus detection in the environment. Ima Fungus, v. 2, n. 2, p. 177-189, 2011.

VENTURA, J. A. Taxonomia de Fusarium e seus segredos: parte I - história, meios e procedimentos de cultivo. Revisão Anual de Patologia de Plantas, v. 7, n. 1, p. 271-298, 1999.

WHITE, T. J. et al. Amplification and direct sequencing of fungal ribosomal RNA genes frophylogenetics. In: INNIS, M. A. et al. (Ed.). PCR Protocols: a guide to methods and applications. New York: Academic Press, 1990. p. 315-322.




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