, 214:7 | Cite as

Marker assisted backcrossing for introgression of Fusarium wilt resistance gene into melon

  • Naeimeh Sousaraei
  • Hossein Ramshini
  • Mahmoud Lotfi
  • Abbas Sharzei


Fusarium wilt, caused by Fusarium oxysporum f. sp. melonis is a common vascular wilt fungal disease in melon across the world. The resistance gene to race 1 of this causal agent, Fom-2, has been previously cloned and its sequence is available. The objective of this research was the introgression of Fom-2 from one resistant (Isabelle) genotype into two susceptible cultivars (Garmak and Tile-torogh) via marker assisted backcrossing. First, the leucine-rich repeats (LRR) domain of Fom-2 from resistant and susceptible genotypes was sequenced to develop functional markers. A length of 1274 bp of the 3′ end of this gene was isolated, cloned and sequenced. The difference between resistant and susceptible genotypes in this region was 28 nucleotide substitutions. Two allele specific primer pairs, Fom2-R409 and Fom2-S253, were designed based on nucleotide substitutions to amplify resistant and susceptible alleles, respectively. For introgression of the gene, donor (Isabelle) and recurrent (Garmak and Tile-torogh) parents were crossed. Resistant plants in BC1F1 and BC2F1 generations were first detected using artificial pathogen inoculation and later the plants were genotyped by functional markers to validate their resistance. The resistant plants were also selected phenotypically in each generation for background genome recovery, which conduced to high similarity of BC3 generation with the recurrent parents. It was proved the developed markers are more precise and efficient than inoculation trial and could be used as confident tools for screening of resistant melon genotypes to Fusarium wilt.


Allele specific PCR Cucumis melo Leucine-rich repeats Marker assisted selection Sequence characterized amplified regions 


  1. Andersen JR, Lubberstedt T (2003) Functional markers in plants. Trends Plant Sci 8:554–560CrossRefPubMedGoogle Scholar
  2. Banihashemi Z (2010) Reaction of Cucumis melo cultivars to races of Fusarium oxysporum f. sp. melonis the cause of melon vascular wilt. Iran J Plant Pathol 46:5–7Google Scholar
  3. Bisognin DA (2002) Origin and evolution of cultivated cucurbits. Ciencia Rural 32:715–723CrossRefGoogle Scholar
  4. Brotman Y, Kovalski ÆI, Dogimont ÆC, Perl-treves R (2005) Molecular markers linked to papaya ring spot virus resistance and Fusarium race 2 resistance in melon. Theor Appl Genet 110(2):337–345. CrossRefPubMedGoogle Scholar
  5. Burger Y, Katzir N, Tzuri G et al (2003) Variation in the response of melon genotypes to Fusarium oxysporum f.sp. melonis race 1 determined by inoculation. Plant Pathol 2:204–211CrossRefGoogle Scholar
  6. Collard BC, Mackill DJ (2008) Marker-assisted selection: an approach for precision plant breeding in the twenty-first century. Philos Trans R Soc Lond B. 363:557–572CrossRefGoogle Scholar
  7. Diaz A, Fergany M, Formisano G et al (2011) A consensus linkage map for molecular markers and Quantitative Trait Loci associated with economically important traits in melon (Cucumis melo L.). BMC Plant Biol 11:111. CrossRefPubMedPubMedCentralGoogle Scholar
  8. Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acids Symp Ser 1999:95–98Google Scholar
  9. Harel-Beja R, Tzuri G, Portnoy V et al (2010) A genetic map of melon highly enriched with fruit quality QTLs and EST markers, including sugar and carotenoid metabolism genes. Theor Appl Genet 121:511–533. CrossRefPubMedGoogle Scholar
  10. Hayashi K, Yoshida H, Ashikawa I (2006) Development of PCR-based allele-specific and InDel marker sets for nine rice blast resistance genes. Theor Appl Genet 113:251–260CrossRefPubMedGoogle Scholar
  11. Jeong HS, Jang S, Han K et al (2015) Marker-assisted backcross breeding for development of pepper varieties (Capsicum annuum) containing capsinoids. Mol Breed 35:1–10. CrossRefGoogle Scholar
  12. Joobeur T, King JJ, Nolin SJ, Thomas CE, Dean RA (2004) The fusarium wilt resistance locus Fom-2 of melon contains a single resistance gene with complex features. Plant J 39:283–297CrossRefPubMedGoogle Scholar
  13. Lau WCP, Rafii MY, Ismail MR et al (2017) Development of advanced fragrant rice lines from MR269 × Basmati 370 through marker-assisted backcrossing. Euphytica 213:11. CrossRefGoogle Scholar
  14. Leida C, Moser C, Esteras C et al (2015) Variability of candidate genes, genetic structure and association with sugar accumulation and climacteric behavior in a broad germplasm collection of melon (Cucumis melo L.). BMC Genet 16:28. CrossRefPubMedPubMedCentralGoogle Scholar
  15. Li Y-H et al (2009) Development of SNP markers and haplotype analysis of the candidate gene for rhg1, which confers resistance to soybean cyst nematode in soybean. Mol Breed 24:63–76CrossRefGoogle Scholar
  16. Madadkhah E, Lotfi M, Nabipour A, Rahmanpour S, Banihashemi Z, Shoorooei M (2012) Enzymatic activities in roots of melon genotypes infected with Fusarium oxysporum f.sp. melonis race 1. Sci Hortic 135:171–176CrossRefGoogle Scholar
  17. McHale L, Tan X, Koehl P, Michelmore RW (2006) Plant NBS-LRR proteins: adaptable guards. Genome Biol 7(4):212. CrossRefPubMedPubMedCentralGoogle Scholar
  18. Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucl Acids Res. 8:4321–4326CrossRefPubMedPubMedCentralGoogle Scholar
  19. Oumouloud A, Mokhtari M, Chikh-Rouhou H et al (2012) Characterization of the Fusarium wilt resistance Fom-2 gene in melon. Mol Breed 30:325–334CrossRefGoogle Scholar
  20. Oumouloud A, El Otmani M, Alvarez J (2015) Molecular characterization of Fom-1 gene and development of functional markers for molecular breeding of resistance to Fusarium race 2 in melon. Euphytica 205:1–11CrossRefGoogle Scholar
  21. Perchepied L, Pitrat M (2004) Polygenic inheritance of partial resistance to Fusarium oxysporum f.sp. melonis race 1.2 in melon. Phytopathology 94:1331–1336CrossRefPubMedGoogle Scholar
  22. Perin C, Hagen L, De Conto V et al (2002) A reference map of Cucumis melo based on two recombinant inbred line populations. Theor Appl Genet 104:1017–1034CrossRefPubMedGoogle Scholar
  23. Risser G, Banihashimi Z, Davis DW (1976) A proposed nomenclature of Fusarium oxysporum f.sp. melonis races and resistance genes in Cucumis melo. Phytopathology 66:1105–1106CrossRefGoogle Scholar
  24. Sherf AF, MacNab AA (1986) Vegetable diseases and their control. Wiley, New YorkGoogle Scholar
  25. Wang Y-H, Thomas CE, Dean RA (2000) Genetic mapping of a fusarium wilt resistance gene (Fom-2) in melon (Cucumis melo L.). Mol Breed 6:379–389CrossRefGoogle Scholar
  26. Wang S, Yang J, Zhang M (2011) Developments of functional markers for Fom-2-mediated fusarium wilt resistance based on single nucleotide polymorphism in melon (Cucumis melo L.). Mol Breed 27:385–393CrossRefGoogle Scholar
  27. Whitaker VM, Bradeen JM, Debener T, Biber A, Hokanson SC (2010) Rdr3, a novel locus conferring black spot disease resistance in tetraploid rose: genetic analysis, LRR profiling, and SCAR marker development. Theor Appl Genet 120:573–585CrossRefPubMedGoogle Scholar
  28. Win KM, Korinsak S, Sirithunya P et al (2013) Marker assisted introgression of multiple genes for bacterial blight resistance into aromatic Myanmar rice MK-75. F Crop Res 154:164–171. CrossRefGoogle Scholar
  29. Yadav PS, Mishra VK, Arun B et al (2015) Enhanced resistance in wheat against stem rust achieved by marker assisted backcrossing involving three independent Sr genes. Curr Plant Biol 2:25–33. CrossRefGoogle Scholar
  30. Yohannes T, Tesfamichael A, Kiambi D et al (2015) Marker-assisted introgression improves Striga resistance in an Eritrean Farmer-Preferred Sorghum Variety. F Crop Res 173:22–29. CrossRefGoogle Scholar
  31. Zheng XY, Wolff DW (2000) Randomly amplified polymorphic DNA markers linked to fusarium wilt resistance in diverse melons. HortScience 35:716–721Google Scholar
  32. Zheng XY, Wolff DW, Baudracco-Arnas S, Pitrat M (1999) Development and utility of cleaved amplified polymorphic sequences (CAPS) and restriction fragment length polymorphisms (RFLPs) linked to the Fom-2 fusarium wilt resistance gene in melon (Cucumis melo L.). Theor Appl Genet 99:453–463CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2017

Authors and Affiliations

  1. 1.Department of HorticultureUniversity of Tehran-Aburaihan campusTehranIran
  2. 2.Deptartment of Agronomy and Plant BreedingUniversity of Tehran-Aburaihan campusTehranIran
  3. 3.University of TehranTehranIran
  4. 4.Deptartment of Entomology and Plant DiseasesUniversity of Tehran-Aburaihan campusTehranIran

Personalised recommendations