Korean Journal of Agricultural Science (농업과학연구)

Institute of Agricultural Science, CNU (충남대학교 농업과학연구소)
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Screening assay for tomato plants resistant to Fusarium oxysporum f. sp. lycopersici race 2 using the expression of the avr2 gene as a selection marker

  • Kim, Mi-Reu (Department of Applied Biology, College of Agriculture & Life Sciences, Chungnam National University) ;
  • Lee, Jeong Jin (Department of Applied Biology, College of Agriculture & Life Sciences, Chungnam National University) ;
  • Min, Jiyoung (Department of Applied Biology, College of Agriculture & Life Sciences, Chungnam National University) ;
  • Kim, Sun Ha (Department of Applied Biology, College of Agriculture & Life Sciences, Chungnam National University) ;
  • Kim, Dae-Gyu (Department of Applied Biology, College of Agriculture & Life Sciences, Chungnam National University) ;
  • Oh, Sang-Keun (Department of Applied Biology, College of Agriculture & Life Sciences, Chungnam National University)
  • Received : 2021.01.20
  • Accepted : 2021.02.05
  • Published : 2021.03.01
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Abstract

Fusarium wilt disease of tomato plants caused by Fusarium oxysporum f.sp. lycopersici (FOL race2) is one of the most important diseases of tomatoes worldwide. In the competition between tomato and FOL, the FOL can win by overcoming the immune system of tomato plants. Resistant interaction between the FOL race2 and tomato plants is controlled by avirulence genes (AVR2) in FOL and the corresponding resistance genes (I2) in tomato plants. In this study, 7 FOL isolates (KACC) were used to test their pathogenicity, and FOL race2 was selected because it is a broad problem in Korea. The Fol40044 isolates showed the most severe pathogenicity, and the avr2 gene was also isolated and identified. Moreover, to select resistance, 20 tomato varieties were inoculated with the Fol40044, and the degree of pathogenicity was evaluated by analyzing the expression of the avr2 gene. As a result, three resistant tomato varieties (PCNUF73, PCNUF101, PCNUF113) were selected, and the expression of the avr2 gene was much lower than that of the control Heinz cultivar. This result shows that the screening assay is very efficient when the avr2 gene is used as a marker to evaluate the expression level when selecting varieties resistant to tomato wilt disease. Based on these results, it is possible to isolate the I2 gene, which exhibits resistance and molecular biological interactions with the AVR2 gene from the three tomato-resistant varieties. The I2 gene provides breeders more opportunities for Fusarium disease resistance and may contribute to our understanding of their interactions with the FOL and host plant.

Keywords

Acknowledgement

이 연구는 충남대학교 연구우수장학금에 의해 지원되었음.

References

  1. Alexander LJ, Tucker CM. 1945. Physiological specialization in the tomato wilt fungus Fusarium oxysporum f. sp. lycopersici. Journal of Agricultural Research 70:303-313.
  2. Amini J, Sidovich D. 2010. The effects of fungicides on Fusarium oxysporum f. sp lycopersici associated with Fusarium wilt of tomato. Journal of Plant Protection Research 50:172-178.
  3. Bohn GW, Tucker CM. 1939. Immunity to Fusarium wilt in the tomato. Science 89:603-604. https://doi.org/10.1126/science.89.2322.603
  4. Cao L, Blekemolen MC, Tintor N, Cornelissen BJC, Takken FLW. 2018. The Fusarium oxysporum Avr2-Six5 effector pair alters plasmodesmatal sxclusion selectivity to facilitate cell-to-cell movement of Avr2. Molecular Plant 11:691-705. https://doi.org/10.1016/j.molp.2018.02.011
  5. Davies JML, 1982. Verticillium and Fusarium wilt of tomato. p. 6. Ministry of Agriculture, Fishery and Food, Northumberland, UK.
  6. De Lamo FJ, Constantin ME, Fresno DH, Boeren S, Rep M, Takken FLW. 2018. Xylem sap proteomics reveals distinct differences Between R gene- and Endophyte-mediated resistance Against Fusarium wilt disease in tomato. Frontiers in Microbiology 9:2977. https://doi.org/10.3389/fmicb.2018.02977
  7. Di X, Gomila J, Ma L, van den Burg HA, Takken FLW. 2016. Uptake of the Fusarium effector Avr2 by tomato is not a cell autonomous event. Front Plant Science 7:1915.
  8. Gabe HL. 1975. Standardization of nomenclature for pathogenic races of Fusarium oxysporum f. sp. lycopersici. Transactions of the British Mycological Society 64:156-159. https://doi.org/10.1016/s0007-1536(75)80089-1
  9. Gawehns F, Cornelissen BJ, Takken FLW. 2013. The potential of effector-target genes in breeding for plant innate immunity. Microbe Biotechnology 6:223-229. https://doi.org/10.1111/1751-7915.12023
  10. Gururani MA, Venkatesh J, Upadhyaya CP, Nookaraju A, Pandey SK, Park SW. 2012. Plant disease resistance genes: Current status and future directions. Physiological and Molecular Plant Pathology 78:51-65. https://doi.org/10.1016/j.pmpp.2012.01.002
  11. Houterman PM, Cornelissen BJ, Rep M. 2008. Suppression of plant resistance gene-based immunity by a fungal effector. PLoS Pathogens 4:e1000061. https://doi.org/10.1371/journal.ppat.1000061
  12. Houterman PM, Ma L, van Ooijen G, de Vroomen MJ, Cornelissen BJ, Takken FLW, Rep M. 2009. The effector protein Avr2 of the xylem-colonizing fungus Fusarium oxysporum activates the tomato resistance protein I-2 intracellularly. Plant Jounal 58:970-978. https://doi.org/10.1111/j.1365-313X.2009.03838.x
  13. Houterman PM, Speijer D, Dekker HL, de Koster CG, Cornelissen BJ, Rep M. 2007. The mixed xylem sap proteome of Fusarium oxysporum-infected tomato plants. Molecular Plant Pathology 8:215-221. https://doi.org/10.1111/j.1364-3703.2007.00384.x
  14. Huang CC, Lindhout P. 1997. Screening for resistance in wild Lycopersicon species to Fusarium oxysporum f. sp. lycopersici race 1 and race 2. Euphytica 93:145-153. https://doi.org/10.1023/A:1002943805229
  15. Hur OS, Ro NY, Ko HC, Kim SG, Rhee JH, Gwag JG, Oh SJ. 2012. Screening for resistance to Fusarium oxysporum f. sp. lycopersici Race 3 using molecular marker in tomato germplasm. Research in Plant Disease 18:304-309. [in Korean] https://doi.org/10.5423/RPD.2012.18.4.304
  16. Jones JB, Jones JP, Stall RE, Zitter TA. 1991. Compendium of tomato diseases. American Phytopathological Society, St. Paul, MNk, USA.
  17. Jones JD, Dangl JL. 2006. The plant immune system. Nature 444:323-329. https://doi.org/10.1038/nature05286
  18. Kim HJ, Lee HR, Hyun JY, Won DC, Hong DO, Cho H, Lee KA, Her NH, Lee JH, Harn CH. 2011. Application of disease resistance markers for developing elite tomato varieties and lines. Horticultural Science & Technology 29:336-344. [in Korean]
  19. Li J, Chitwood J, Menda N, Mueller L, Hutton SF. 2018. Linkage between the I-3 gene for resistance to Fusarium wilt race 3 and increased sensitivity to bacterial spot in tomato. Theory Apply Genetics 131:145-155. https://doi.org/10.1007/s00122-017-2991-4
  20. Ma L, Cornelissen BJ, Takken FLW. 2013. A nuclear localization for Avr2 from Fusarium oxysporum is required to activate the tomato resistance protein I-2. Front Plant Science 4:94. https://doi.org/10.3389/fpls.2013.00094
  21. Ma L, Houterman PM, Gawehns F, Cao L, Sillo F, Richter H. 2015. The AVR2- SIX5 gene pair is required to activate I-2-mediated immunity in tomato. New Phytology 208:507-518. https://doi.org/10.1111/nph.13455
  22. McGrath DJ, Gillespie D, Vawdrey L. 1987. Inheritance of resistance to Fusarium oxysporum f. sp. lycopersici races 2 and 3 in Lycopersicon pennellii. Australian Journal of Agricultural Research 38:729-733. https://doi.org/10.1071/AR9870729
  23. Mes JJ. 1999. Molecular aspects of the interaction between tomato and Fusarium oxysporum f. sp. lycopersici. Ph.D. Thesis, University of Amsterdam, Amsterdam, Netherlands.
  24. Michielse CB, Rep M. 2009. Pathogen profile update: Fusarium oxysporum. Molecular Plant Pathology 10:311-324. https://doi.org/10.1111/j.1364-3703.2009.00538.x
  25. Park GR, Jang HA, Jo SH, Park Y, Oh SK, Nam M. 2018. Development of SNP marker set for marker-assisted backcrossing (MABC) in cultivating tomato varieties. Korean Journal of Agricultural Science 45:385-400. [in Korean] https://doi.org/10.7744/KJOAS.20180061
  26. Park JS. 1958. Fungus disease of plants in Korea (I). College of Agric., Chungnam National University Bulletin 1:62. [In Korean]
  27. Park MS, Jang KS, Choi YH, Kim JC, Choi GJ. 2013. Simple mass-screening methods for resistance of tomato to Fusarium oxysporum f. sp. lycopersici. Horticultural Science and Technology 31:110-116. [in Korean] https://doi.org/10.7235/hort.2013.12134
  28. RDA (Rural Development Administration). 2020. Tomato. RDA, Suwon, Korea. [in Korean]
  29. Rep M, Meijer M, Houterman PM, van der Does HC, Cornelissen BJ. 2005. Fusarium oxysporum evades I-3-mediated resistance without altering the matching avirulence gene. Molecular Plant Microbe Interaction 18:15-23. https://doi.org/10.1094/MPMI-18-0015
  30. Rep M, van der Does HC, Meijer M, van Wijk R, Houterman PM. 2004. A small, cysteine-rich protein secreted by Fusarium oxysporum during colonization of xylem vessels is required for I-3-mediated resistance in tomato. Mol Microbiol 53:1373-1383. https://doi.org/10.1111/j.1365-2958.2004.04177.x
  31. Saitoh K, Togashi K, Arie T, Teraoka T. 2006. A simple method for a mini preparation of fungal DNA. Journal of General Plant Pathology 72:348-350. https://doi.org/10.1007/s10327-006-0300-1
  32. Schmidt SM, Houterman PM, Schreiver I, Ma L, Amyotte S, Chellappan B, Boeren S, Takken FLW, Rep M. 2013. MITEs in the promoters of effector genes allow prediction of novel virulence genes in Fusarium oxysporum. BMC Genomics 14:119. https://doi.org/10.1186/1471-2164-14-119
  33. Simons G, Groenendijk J, Wijbrandi J, Reijans M, Groenen J, Diergaarde P, Van der Lee T, Bleeker M, Onstenk J, de Both M. 1998. Dissection of the fusarium I2 gene cluster in tomato reveals six homologs and one active gene copy. Plant Cell 10:1055-1068. https://doi.org/10.2307/3870690
  34. Takken FLW, Suzan RL, GabrieEls HEJ, Westerink N, Lu R, de Wit PJGM, Joosten MHAJ. 2000. A functional cloning strategy, based on a binary PVX-expression vector, to isolate HR-inducing cDNAs of plant pathogens. The Plant Journal 24:275-283. https://doi.org/10.1046/j.1365-313x.2000.00866.x
  35. Takken F, Rep M. 2010. The arms race between tomato and Fusarium oxysporum. Molecular Plant Pathology 11:309-314. https://doi.org/10.1111/j.1364-3703.2009.00605.x
  36. van der Does M, Knippertz P, Zschenderlein P, Giles Harrison R, Stuut JBW. 2018. The mysterious long-range transport of giant mineral dust particles. Science Advances 4:eaau2768. https://doi.org/10.1126/sciadv.aau2768