The Plant Pathology Journal

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

DOI QR Code

Development qRT-PCR Protocol to Predict Strawberry Fusarium Wilt Occurrence

  • Hong, Sung Won (Division of Applied Life Science (BK21Plus), Gyeongsang National University) ;
  • Kim, Da-Ran (Department of Plant Medicine, Institute of Agriculture & Life Science, Gyeongsang National University) ;
  • Kim, Ji Su (Division of Applied Life Science (BK21Plus), Gyeongsang National University) ;
  • Cho, Gyeongjun (Division of Applied Life Science (BK21Plus), Gyeongsang National University) ;
  • Jeon, Chang Wook (Division of Applied Life Science (BK21Plus), Gyeongsang National University) ;
  • Kwak, Youn-Sig (Division of Applied Life Science (BK21Plus), Gyeongsang National University)
  • Received : 2017.12.23
  • Accepted : 2018.01.30
  • Published : 2018.06.01
Download PDF
⟨ Previous Next ⟩

Abstract

Strawberry Fusarium wilt disease, caused by Fusarium oxysporum f. sp. fragariae, is the most devastating disease in strawberry production. The pathogen produces chlamydospores which tolerate against harsh environment, fungicide and survive for decades in soil. Development of detection and quantification techniques are regarded significantly in many soilborne pathogens to prevent damage from diseases. In this study, we improved specific-quantitative primers for F. oxysporum f. sp. fragariae to reveal correlation between the pathogen density and the disease severity. Standard curve $r^2$ value of the specific-quantitative primers for qRT-PCR and meting curve were over 0.99 and $80.5^{\circ}C$, respectively. Over pathogen $10^5cfu/g$ of soil was required to cause the disease in both lab and field conditions. With the minimum density to develop the wilt disease, the pathogen affected near 60% in nursery plantation. A biological control microbe agent and soil solarization reduced the pathogen population 2-fold and 1.5-fold in soil, respectively. The developed F. oxysporum f. sp. fragariae specific qRT-PCR protocol may contribute to evaluating soil healthiness and appropriate decision making to control the disease.

Keywords

References

  1. Anaya, N. and Roncero, M. I. G. 1995. Skippy, a retrotransposon from the fungal plant pathogen Fusarium oxysporum. Mol. Gen. Genet. 249:637-647. https://doi.org/10.1007/BF00418033
  2. Ashworth, L. and Gaona, S. 1982. Evaluation of clear polyethylene mulch for controlling Verticillium wilt in established pistachio nut groves. Phytopathology 72:243-246. https://doi.org/10.1094/Phyto-72-243
  3. Barrett, L. G., Kniskern, J. M., Bodenhausen, N., Zhang, W. and Bergelson, J. 2009. Continua of specifcity and virulence in plant host-pathogen interactions: causes and consequences. New Phytol. 183:513-529. https://doi.org/10.1111/j.1469-8137.2009.02927.x
  4. Bilodeau, G. J., Koike, S. T., Uribe, P. and Martin, F. N. 2012. Development of an assay for rapid detection and quantifcation of Verticillium dahliae in soil. Phytopathology 102:331-343. https://doi.org/10.1094/PHYTO-05-11-0130
  5. Bonants, P. J., van Gent-Pelzer, M. P., Hooftman, R., Cooke, D. E., Guy, D. C. and Duncan, J. M. 2004. A combination of baiting and different PCR formats, including measurement of real-time quantitative fuorescence, for the detection of Phytophthora fragariae in strawberry plants. Eur. J. Plant Pathol. 110:689-702. https://doi.org/10.1023/B:EJPP.0000041551.26970.0e
  6. Capote, N., Pastrana, A. M., Aguado, A. and Sanchez-Torres, P. 2012. Molecular tools for detection of plant pathogenic fungi and fungicide resistance. In: Plant pathology, ed. by C. J. R. Cumagun, pp. 151-202. InTech, Rijeka, Croatia.
  7. Cha, J. Y., Han, S., Hong, H. J., Cho, H., Kim, D., Kwon, Y., Kwon, S. K., Crusemann, M., Lee, Y. B., Kim, J. F., Giaever, G., Nislow, C., Moore, B. S., Thomashow, L. S., Weller, D. M. and Kwak, Y. S. 2016. Microbial and biochemical basis of a Fusarium wilt-suppressive soil. ISME J. 10:119-129. https://doi.org/10.1038/ismej.2015.95
  8. Chalvet, F., Grimaldi, C., Kaper, F., Langin, T. and Daboussi, M. 2003. Hop, an active mutator-like element in the genome of the fungus Fusarium oxysporum. Mol. Biol. Evol. 20:1362-1375. https://doi.org/10.1093/molbev/msg155
  9. Filion, M., Arnaud, M. and Jabaji-Hare, S. 2003. Quantification of Fusarium solani f. sp. phaseoli in mycorrhizal bean plants and surrounding mycorrhizosphere soil using real-time polymerase chain reaction and direct isolations on selective media. Phytopathology 93:229-235. https://doi.org/10.1094/PHYTO.2003.93.2.229
  10. Frank, S. A. 1996. Models of parasite virulence. Q. Rev. Biol. 71:37-78. https://doi.org/10.1086/419267
  11. Hua-Van, A., Daviere, J., Kaper, F., Langin, T. and Daboussi, M. 2000. Genome organization in Fusarium oxysporum: clusters of class II transposons. Curr. Genet. 37:339-347. https://doi.org/10.1007/s002940050537
  12. Jimenez-Fernandez, D., Montes-Borrego, M., Navas-Cortes, J. A., Jimenez-Diaz, R. M. and Landa, B. B. 2010. Identification and quantifcation of Fusarium oxysporum in planta and soil by means of an improved specifc and quantitative PCR assay. Appl. Soil Ecol. 46:372-382. https://doi.org/10.1016/j.apsoil.2010.10.001
  13. Katan, J. 1981. Solar heating (solarization) of soil for control of soilborne pests. Annu. Rev. Phytopathol. 19:211-236. https://doi.org/10.1146/annurev.py.19.090181.001235
  14. Kim, D., Gang, G., Cho, H., Yoon, H. and Kwak, Y. 2015. Evaluation of antimicrobial activity and disease control effcacy of sodium dichloroisocyanurate (NaDCC) against major strawberry diseases. Korean J. Pestic. Sci. 19:47-53 (in Korean). https://doi.org/10.7585/kjps.2015.19.1.47
  15. Kodama, T. 1974. Characters of strawberry yellows caused by Fusarium and difference of its effect on the grown varieties. Bull. Nara Agri. Expt. Sta. 6:68-75.
  16. Kodama, T. and Fukuit, T. 1982. Application of solar heating with plastic-flm mulching in the out-door feld for control of Fusarium wilt of strawberry. Japanese J. Phytopathol. 48:699-701. https://doi.org/10.3186/jjphytopath.48.699
  17. Li, M., Asano, T., Suga, H. and Kageyama, K. 2011. A multiplex PCR for the detection of Phytophthora nicotianae and P. cactorum, and a survey of their occurrence in strawberry production areas of Japan. Plant Dis. 95:1270-1278. https://doi.org/10.1094/PDIS-01-11-0076
  18. Lievens, B., Brouwer, M., Vanachter, A. C., Cammue, B. P. and Thomma, B. P. 2006. Real-time PCR for detection and quantifcation of fungal and oomycete tomato pathogens in plant and soil samples. Plant Sci. 171:155-165. https://doi.org/10.1016/j.plantsci.2006.03.009
  19. Lievens, B., Brouwer, M., Vanachter, A. C., Levesque, C. A., Cammue, B. and Thomma, B. P. 2005. Quantitative assessment of phytopathogenic fungi in various substrates using a DNA macroarray. Environ. Microbial. 7:1698-1710. https://doi.org/10.1111/j.1462-2920.2005.00816.x
  20. Lievens, B., Rep, M. and Thomma, B. P. 2008. Recent developments in the molecular discrimination of formae speciales of Fusarium oxysporum. Pest Manag. Sci. 64781-788. https://doi.org/10.1002/ps.1564
  21. Maas, J. L. 1998. Compendium of strawberry diseases. APS press, St. Paul, MN, USA.
  22. McCartney, H. A., Foster, S. J. and Fraaije, B. A. and Ward, E. 2003. Molecular diagnostics for fungal plant pathogens. Pest Manag. Sci. 59:129-142. https://doi.org/10.1002/ps.575
  23. Nagaraian, G., Nam, M., Song, J., Yoo, S. and Kim, H. 2004. Genetic variation in Fusarium oxysporum f. sp. fagariae populations based RAPD and rDNA RFLP analyses. Plant Pathol. J. 20:264-270. https://doi.org/10.5423/PPJ.2004.20.4.264
  24. Pasquali, M., Acquadro, A., Balmas, V., Migheli, Q., Gullino, M. L. and Garibaldi, A. 2004. Development of PCR primers for a new Fusarium oxysporum pathogenic on Paris daisy (Argyranthemum frutescens L.). Eur. J. Plant Pathol. 110:7-11.
  25. Pinkerton, J., Ivors, K., Miller, M. and Moore, L. 2000. Effect of soil solarization and cover crops on populations of selected soilborne plant pathogens in western Oregon. Plant Dis. 84:952-960. https://doi.org/10.1094/PDIS.2000.84.9.952
  26. Porebski, S., Bailey, L. G. and Baum, R. 1997. Modifcation of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components. Plant Mol. Biol. Rep. 15:8-15. https://doi.org/10.1007/BF02772108
  27. Raio, A., Zoina, A. and Moore, L. 1997. The effect of solar heating of soil on natural and inoculated agrobacteria. Plant Pathol. 46:320-328. https://doi.org/10.1046/j.1365-3059.1997.d01-28.x
  28. Sankaran, S., Mishra, A., Ehsani, R. and Davis, C. 2010. A review of advanced techniques for detecting plant diseases. Comput. Electron. Agric. 72:1-13. https://doi.org/10.1016/j.compag.2010.02.007
  29. Scarlett, K., Tesoriero, L., Daniel, R. and Guest, D. 2013. Detection and quantification of Fusarium oxysporum f. sp. cuc- umerinum in environmental samples using a specifc quantitative PCR assay. Eur. J. Plant Pathol. 137:315-324. https://doi.org/10.1007/s10658-013-0244-1
  30. Schena, L., Li Destri, N. M., Sanzani, S., Faedda, R., Ippolito, A. and Cacciola, S. 2013. Development of quantitative PCR detection methods for phytopathogenic fungi and oomycetes. J. Plant Pathol. 95:7-24.
  31. Suga, H., Hirayama, Y., Morishima, M., Suzuki, T., Kageyama, K. and Hyakumachi, M. 2013. Development of PCR primers to identify Fusarium oxysporum f. sp. fragariae. Plant Dis. 97:619-625. https://doi.org/10.1094/PDIS-07-12-0663-RE
  32. Tezuka, N. and Makino, T. 1991. Biological control of Fusarium wilt of strawberry by nonpathogenic Fusarium oxysporum isolated from strawberry. Japanese J. Phytopathol. 57:506-511. https://doi.org/10.3186/jjphytopath.57.506
  33. van Dam, P., Fokkens, L., Schmidt, S. M., Linmans, J. H., Kistler, H. C., Ma, L. and Rep, M. 2016. Effector profles distinguish formae speciales of Fusarium oxysporum. Environ. Microbiol. 18:4087-4102. https://doi.org/10.1111/1462-2920.13445
  34. Winks, B. and Williams, Y. 1965. A wilt of strawberry caused by a new form of Fusarium oxysporum. Queensland J. Agric. Anim. Sci. 22:475-479.
  35. Zambounis, A., Paplomatas, E. and Tsaftaris, A. 2007. Intergenic spacer-RFLP analysis and direct quantifcation of Australian Fusarium oxysporum f. sp. vasinfectum isolates from soil and infected cotton tissues. Plant Dis. 91:1564-1573. https://doi.org/10.1094/PDIS-91-12-1564