The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
권호 표지
DOI QR코드

DOI QR Code

Construction of a System for the Strawberry Nursery Production towards Elimination of Latent Infection of Anthracnose Fungi by a Combination of PCR and Microtube Hybridization

  • Furuta, Kazuyoshi (Agroscience Research Laboratories, Plant Biotechnology Center, Hokusan Co., Ltd.) ;
  • Nagashima, Saki (Research Faculty of Agriculture, Hokkaido University) ;
  • Inukai, Tsuyoshi (Research Faculty of Agriculture, Hokkaido University) ;
  • Masuta, Chikara (Research Faculty of Agriculture, Hokkaido University)
  • Received : 2016.05.27
  • Accepted : 2016.10.04
  • Published : 2017.02.01
Download PDF
⟨ Previous Next ⟩

Abstract

One of the major problems in strawberry production is difficulty in diagnosis of anthracnose caused by Colletotrichum acutatum or Glomerella cingulata in latent infection stage. We here developed a diagnostic tool for the latent infection consisting of initial culturing of fungi, DNA extraction, synthesis of PCR-amplified probes and microtube hybridization (MTH) using a macroarray. The initial culturing step is convenient to lure the fungi out of the plant tissues, and to extract PCR-inhibitor-free DNA directly from fungal hyphae. For specific detection of the fungi, PCR primers were designed to amplify the fungal MAT1-2 gene. The subsequent MTH step using the PCR products as probes can replace the laborious electrophoresis step providing us sequence information and high-throughput screening. Using this method, we have conducted a survey for a few thousands nursery plants every year for three consecutive years, and finally succeeded in eliminating latent infection in the third year of challenge.

Keywords

References

  1. Curry, K. J., Abril, M., Avant, J. B. and Smith, B. J. 2002. Strawberry anthracnose: histopathology of Colletotrichum acutatum and C. fragariae. Phytopathology 92:1055-1063. https://doi.org/10.1094/PHYTO.2002.92.10.1055
  2. Denoyes-Rothan, B., Guerin, G., Lerceteau-Kohler, E. and Risser, G. 2005. Inheritance of resistance to Colletotrichum acutatum in Fragaria x ananassa. Phytopathology 95:405-412. https://doi.org/10.1094/PHYTO-95-0405
  3. Du, M., Schardl, C. L., Nuckles, E. M. and Vaillancourt, L. J. 2005. Using mating-type gene sequences for improved phylogenetic resolution of Collectotrichum species complexes. Mycologia 97:641-658. https://doi.org/10.1080/15572536.2006.11832795
  4. Freeman, S., Horowitz, S. and Sharon, A. 2001. Pathogenic and nonpathogenic lifestyles in Colletotrichum acutatum from strawberry and other plants. Phytopathology 91:986-992. https://doi.org/10.1094/PHYTO.2001.91.10.986
  5. Freeman, S. and Katan, T. 1997. Identification of Colletotrichum species responsible for anthracnose and root necrosis of strawberry in Israel. Phytopathology 87:516-521. https://doi.org/10.1094/PHYTO.1997.87.5.516
  6. Garrido, C., Carbu, M., Fernandez-Acero, F. J., Boonham, N., Colyer, A., Cantoral, J. M. and Budge, G. 2009. Development of protocols for detection of Colletotrichum acutatum and monitoring of strawberry anthracnose using real-time PCR. Plant Pathol. 58:43-51. https://doi.org/10.1111/j.1365-3059.2008.01933.x
  7. Kikuchi, M., Ogawara, T., Hashimoto, Y., Miyamoto, T., Kaneda, M. and Tomita, Y. 2010. Identification of Colletotrichum species causing strawberry anthracnose and distribution of fungal strains resistant to some fungicides in Ibaraki prefecture. Bull. Hortic. Inst. Ibaraki Agric. Cent. 17:35-42 (in Japanese).
  8. Kuchta, P., Jecz, T. and Korbin, M. 2008. The suitability of PCR-based techniques for detecting Verticillium dahliae in strawberry plants and soil. J. Fruit Ornam. Plant Res. 16: 295-304.
  9. Leandro, L. F., Gleason, M. L., Nutter, F. W., Wegulo, S. N. and Dixon, P. M. 2001. Germination and sporulation of Colletotrichum acutatum on symptomless strawberry leaves. Phytopathology 91:659-664. https://doi.org/10.1094/PHYTO.2001.91.7.659
  10. Legard, D. E. 2000. Colletotrichum diseases of strawberry in Florida. In: Colletotrichum: host specificity, pathology, and host-pathogen interaction, eds. by D. Prusky, S. Freeman and M. B. Dickman, pp. 292-299. APS Press, St. Paul, MN, USA.
  11. Lerceteau-Kohler, E., Guerin, G. and Denoyes-Rothan, B. 2005. Identification of SCAR markers linked to Rca2 anthracnose resistance gene and their assessment in strawberry germplasm. Theor. Appl. Genet. 111:862-870. https://doi.org/10.1007/s00122-005-0008-1
  12. MacKenzie, S. J., Legard, D. E., Timmer, L. W., Chandler, C. K. and Peres, N. A. 2006. Resistance of strawberry cultivars to crown rot caused by Colletotrichum gloeosporioides isolates from Florida is nonspecific. Plant Dis. 90:1091-1097. https://doi.org/10.1094/PD-90-1091
  13. Martinez-Culebras, P. V., Barrio, E., Garcia, M. D. and Querol, A. 2000. Identification of Colletotrichum species responsible for anthracnose of strawberry based on the internal transcribed spacers of the ribosomal region. FEMS Microbiol. Lett. 189:97-101. https://doi.org/10.1111/j.1574-6968.2000.tb09213.x
  14. Parikka, P. and Lemmetty, A. 2004. Tracing latent infection of Colletotrichum acutatum on strawberry by PCR. Eur. J. Plant Pathol. 110:393-398. https://doi.org/10.1023/B:EJPP.0000021073.67137.d2
  15. Shieh, J., To, C., Carramao, J., Nishimura, N., Maruta, Y., Hashimoto, Y., Wright, D., Wu, H. C. and Azarani, A. 2002. High-throughput array production using precision glass syringes. Biotechniques 32:1360-1362, 1364-1365.
  16. Shimura, H., Furuta, K. and Masuta, C. 2015. Detection of plant viruses in mixed infection by macroarray-assisted method. In: Plant virology protocols: new approaches to detect viruses and host responses, eds. by I. Uyeda and C. Masuta, pp. 1-11. Humana Press, New York, NY, USA.
  17. Smith, B. J. and Black, L. L. 1990. Morphological, cultural, and pathogenic variation among Colletotrichum species isolated from strawberry. Plant Dis. 74:69-76. https://doi.org/10.1094/PD-74-0069
  18. Sreenivasaprasad, S., Shardada, K., Brown, A. E. and Mills, P. R. 1996. PCR-based detection of Colletotrichum acutatum on strawberry. Plant Pathol. 45:650-655. https://doi.org/10.1046/j.1365-3059.1996.d01-3.x
  19. Sugiyama, S., Masuta, C., Sekiguchi, H., Uehara, T., Shimura, H. and Maruta, Y. 2008. A simple, sensitive, specific detection of mixed infection of multiple plant viruses using macroarray and microtube hybridization. J. Virol. Methods 153:241-244. https://doi.org/10.1016/j.jviromet.2008.07.028
  20. Sumino, A., Shinmura, A., Narimatsu, Y., Matsui, R. and Nishimura, S. 2009. Occurrence of strawberry anthracnose caused by Glomerella cingulata (Colletotrichum gloeosporioides) in Hokkaido. Annu. Rep. Soc. Plant Prot. North Jpn. 60:82-87 (in Japanese).
  21. Urena-Padilla, A. R., MacKenzie, S. J., Bowen, B. W. and Legard, D. E. 2002. Etiology and population genetics of Colletotrichum spp. causing crown and fruit rot of strawberry. Phytopathology 92:1245-1252. https://doi.org/10.1094/PHYTO.2002.92.11.1245
  22. White, T. J., Bruns, T., Lee, S. and Taylor, J. W. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: PCR protocols: a guide to methods and applications, ed. by M. A. Innis, pp. 315-322. Academic Press, San Diego, CA, USA.