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Molecular characterization of Fom-1 gene and development of functional markers for molecular breeding of resistance to Fusarium race 2 in melon

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Abstract

Melon Fusarium wilt disease caused by the soil-borne pathogen Fusarium oxysporum f. sp. melonis (Fom) is one of the most devastating melon diseases worldwide. Recently, the Fom-1 gene responsible for resistance against Fom races 0 and 2 was cloned. In this study we amplified, cloned and sequenced full genomic DNA and cDNA of Fom-1 from several melon resistant and susceptible accessions using three pairs of primers designed within this gene. Sequence analyses showed that this gene contains four exons interrupted by three introns. The comparative sequence analysis of the cloned cDNA amplicons from resistant and susceptible genotypes revealed eight nucleotide substitutions, within Fom-1 coding regions, among which four were non-synonymous. RT-PCR revealed that the Fom-1 expression is induced by Fom race 2 inoculation. The Fom-1 predicted protein (FOM-1) exhibits a tripartite modular structure composed of an N-terminal TIR domain, a central NB–ARC domain and a C-terminal LRR domain. FOM-1 from resistant melon accessions share four amino acid differences relative to the FOM-1 protein in susceptible ones. Two amino acid substitutions N56K and R103H were located at the FOM-1 TIR domain and the substitution E385K in the NB–ARC domain. Based on single nucleotide polymorphisms within the coding region of the Fom-1 locus, we have generated two CAPS markers, Fom-1R and Fom-1S. Results from screening various melon accessions clearly demonstrated the usefulness of both functional CAPS markers in the marker-assisted selection for melon breeding programs.

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References

  • Anderson PA, Lawrence GJ, Morrish BC, Ayliffe MA, Finnegan EJ, Ellis JG (1997) Inactivation of the flax rust resistance gene M associated with loss of a repeated unit within the leucine-rich repeat coding region. Plant Cell 9:641–651

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Bernoux M, Ve T, Williams S, Warren C, Hatters D, Valkov E, Zhang X, Ellis JG, Kobe B, Dodds PN (2011) Structural and functional analysis of a plant resistance protein TIR domain reveals interfaces for self-association, signaling, and autoregulation. Cell Host Microbe 9:200–211

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Brotman Y, Kovalski I, Dogimont C, Pitrat M, Portnoy V, Perl-Treves R (2005) Molecular markers linked to papaya ring spot virus resistance and Fusarium race 2 resistance in melon. Theor Appl Genet 110:337–345

    Article  CAS  PubMed  Google Scholar 

  • Brotman Y, Normantovich M, Goldenberg Z, Zvirin Z, Kovalski I, Stovbun N, Donniger T, Bolger AM, Troadec C, Bendahmane A, Cohen R, Katzir N, Pitrat M, Dogimont C, Perl-Treves R (2013) Dual resistance of melon to Fusarium oxysporum races 0 and 2 and to Papaya ring-spot virus is controlled by a pair of head-to-head oriented NB–LRR genes of unusual architecture. Mol Plant 6:235–238

    Article  CAS  PubMed  Google Scholar 

  • Chen X, Shang J, Chen D, Lei C, Zou Y, Zhai W, Liu G, Xu J, Ling Z, Cao G, Ma B, Wang Y, Zhao X, Li S, Zhu L (2006) A B-lectin receptor kinase gene conferring rice blast resistance. Plant J 46:794–804

    Article  CAS  PubMed  Google Scholar 

  • Dinesh-Kumar SP, Baker BJ (2000) Alternatively spliced N resistance gene transcripts: their possible role in tobacco mosaic virus resistance. Proc Natl Acad Sci USA 97:1908–1913

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Dodds PN, Lawrence GJ, Ellis JG (2001) Six amino acid changes confined to the leucine-rich repeat beta-strand/beta-turn motif determine the difference between the P and P2 rust resistance specificities in flax. Plant Cell 13:163–178

    CAS  PubMed Central  PubMed  Google Scholar 

  • Doyle JJ, Doyle JL (1987) A rapid isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15

    Google Scholar 

  • Fradin EF, Zhang Z, Ayala JCJ, Castroverde CDM, Nazar RN, Robb J, Liu CM, Thomma BPHJ (2009) Genetic dissection of Verticillium wilt resistance mediated by tomato Ve1. Plant Physiol 150:320–332

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Garcia-Mas J, Leeuwen HV, Monfort A, Carmen de Vicente M, Puigdomenech P, Arus P (2001) Cloning and mapping of resistance gene homologues in melon. Plant Sci 161:165–172

    Article  CAS  Google Scholar 

  • Garcia-Mas J et al. (2012) The genome of melon (Cucumis melo L.). Proc Natl Acad Sci USA 109:11872–11877

  • Hammond-Kosack KE, Jones JDG (1997) Plant disease resistance genes. Annu Rev Plant Physiol Plant Mol Biol 48:575–607

    Article  CAS  PubMed  Google Scholar 

  • 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 39:283–297

    Article  CAS  PubMed  Google Scholar 

  • Jupe F, Pritchard L, Etherington GJ, Mackenzie K, Cock PJ, Wright F, Kumar Sharma S, Bolser D, Bryan GJ, Jones JD, Hein I (2012) Identification and localization of the NB–LRR gene family within the potato genome. BMC Genom 13:75

    Article  CAS  Google Scholar 

  • Kar B, Nanda S, Nayak PK, Nayak S, Joshi RK (2013) Molecular characterization and functional analysis of CzR1, a coiledcoil-nucleotide-binding-site-leucine-rich repeat R-gene from Curcuma zedoaria Loeb. that confers resistance to Pythium aphanidermatum. Physiol Mol Plant Pathol 83:59–68

    Article  CAS  Google Scholar 

  • Lawrence GJ, Finnegan EJ, Ayliffe MA, Ellis JG (1995) The L6 gene for flax rust resistance is related to the Arabidopsis bacterial resistance gene RPS2 and tobacco viral resistance gene N. Plant Cell 7:1195–1206

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Leister D, Kurth J, Laurie DA, Yano M, Sasaki T, Devos K, Graner A, Schulze-Lefert P (1998) Rapid reorganization of resistance gene homologues in cereal genomes. Proc Natl Acad Sci USA 95:370–375

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Lukasik-Shreepaathy E, Slootweg E, Richter H, Goverse A, Cornelissen BJC, Takken FLW (2012) Dual regulatory roles of the extended N terminus for Activation of the tomato Mi-1.2 resistance protein. Mol Plant-Microbe Interact 25:1045–1057

    Article  CAS  PubMed  Google Scholar 

  • Mes J, Van Doorn A, Wijbrandi J, Simons G, Cornelissen B, Haring M (2000) Expression of the Fusarium resistance gene I-2 co-localizes with the site of fungal containment. Plant J 23:183–193

    Article  CAS  PubMed  Google Scholar 

  • Meyers BC, Kozik A, Griego A, Kuang H, Michelmore RW (2003) Genome-wide analysis of NBS–LRR-encoding genes in Arabidopsis. Plant Cell 15:809–834

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Oumouloud A, Arnedo-Andres MS, Gonzalez-Torres R, Álvarez JM (2008) Development of molecular markers linked to the Fom-1 locus for resistance to Fusarium race 2 in melon. Euphytica 164:347–356

    Article  CAS  Google Scholar 

  • Oumouloud A, Arnedo-Andres MS, González-Torres R, Álvarez JM (2009) Morphological and molecular characterization of melon accessions resistant to Fusarium wilts. Euphytica 169:69–79

    Article  Google Scholar 

  • Oumouloud A, Arnedo-Andrés MS, González-Torres R, Álvarez JM (2010) Inheritance of resistance to Fusarium oxysporum f. sp. melonis races 0 and 2 in melon accession Tortuga. Euphytica 176:183–189

    Article  Google Scholar 

  • Oumouloud A, Mokhtari M, Chikh-Rouhou H, Arnedo-Andrés MS, González-Torres R, Álvarez JM (2012) Characterization of the Fusarium wilt resistance Fom-2 gene in melon. Mol Breed 30:325–334

    Article  CAS  Google Scholar 

  • Oumouloud A, El-Otmani M, Chikh-Rouhou H, Garcés-Claver A, González-Torres R, Perl-Treves R, Alvarez JM (2013) Breeding melon for resistance to Fusarium wilt: recent developments. Euphytica 192:155–169

    Article  CAS  Google Scholar 

  • Risser G, Banihashimi Z, Davis DW (1976) A proposed nomenclature of Fusarium oxysporum f. sp. melonis races and résistance genes in Cucumis melo. Phytopathology 66:1105–1106

    Article  Google Scholar 

  • Rozen S, Skaletsky H (2000) Primer3 on the www for general users and for biologist programmers. In: Krawetz SA, Misener S (eds) Bioinformatics methods and protocols: methods in molecular biology, vol 132. Humana Press, Totowa, pp 365–386

    Chapter  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Press, Plainview

    Google Scholar 

  • Simpson GG, Filipowicz W (1996) Splicing of precursors to mRNA in higher plants: mechanism, regulation, and subnuclear organization of the spliceosomal machinery. Plant Mol Biol 32:1–41

    Article  CAS  PubMed  Google Scholar 

  • Song J, Bradeen JM, Naess SK, Raasch JA, Wielgus SM, Haberlach GT, Liu J, Kuang H, Austin-Phillips S, Buell CR, Helgeson JP, Jiang J (2003) Gene RB cloned from Solanum bulbocastanum confers broad spectrum resistance to potato late blight. Proc Natl Acad Sci USA 100:9128–9133

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Tan X, Meyers BC, Kozik A, West M, Morgante M, Clair DA, Bent AF, Michelmore RW (2007) Global expression analysis of nucleotide binding site-leucine rich repeat-encoding and related genes in Arabidopsis. BMC Plant Biol 7:56

    Article  PubMed Central  PubMed  Google Scholar 

  • Tezuka T, Waki K, Yashiro K, Kuzuya M, Ishikawa T, Takatsu Y, Miyagi M (2009) Construction of a linkage map and identification of DNA markers linked to Fom-1, a gene conferring resistance to Fusarium oxysporum f.sp. melonis race 2 in melon. Euphytica 168:177–188

    Article  CAS  Google Scholar 

  • Tezuka T, Waki K, Kuzuya M, Ishikawa T, Takatsu Y, Miyagi M (2011) Development of new DNA markers linked to the Fusarium wilt resistance locus Fom-1 in melon. Plant Breed 130:261–267

    Article  CAS  Google Scholar 

  • Van Leeuwen H, Garcia-Mas J, Coca M, Puigdoménech P, Monfort A (2005) Analysis of the melon genome in regions encompassing TIR–NBS–LRR resistance genes. Mol Gen Genomics 273:240–251

    Article  CAS  Google Scholar 

  • Van Ooijen G, Mayr G, Albrecht M, Cornelissen BJ, Takken FL (2008a) Transcomplementation, but not physical association of the CC–NB–ARC and LRR domains of tomato R protein Mi-1.2 is altered by mutations in the ARC2 subdomain. Mol Plant 1:401–410

    Article  PubMed  Google Scholar 

  • Van Ooijen G, Mayr G, Kasiem MM, Albrecht M, Cornelissen BJ, Takken FL (2008b) Structure-function analysis of the NB–ARC domain of plant disease resistance proteins. J Exp Bot 59:1383–1397

    Article  PubMed  Google Scholar 

  • 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 27:385–393

    Article  CAS  Google Scholar 

  • Wang X, Xu B, Zhao L, Gao P, MA H, Luan F (2014) Expression analysis of Fusarium wilt resistance gene in melon by real-time quantitative PCR. Pak J Bot 46:713–717

    CAS  Google Scholar 

  • Whitham S, Dinesh-Kumar SP, Choi D, Hehl R, Corr C, Baker B (1994) The product of the tobacco mosaic virus resistance gene N: similarity to toll and the interleukin-1 receptor. Cell 78:1101–1115

    Article  CAS  PubMed  Google Scholar 

  • Wladimir IL, Tameling JH, Vossen MA, Lengauer T, Berden JA, Haring MA, Cornelissen BJC, Takken FLW (2006) Mutations in the NB–ARC Domain of I-2 that impair ATP hydrolysis cause autoactivation. Plant Physiol 140:1233–1245

    Article  Google Scholar 

  • Xiang Y, Cao Y, Xu C, Li X, Wang S (2006) Xa3, conferring resistance for rice bacterial blight and encoding a receptor kinase-like protein, is the same as Xa26. Theor Appl Genet 113:1347–1355

    Article  CAS  PubMed  Google Scholar 

  • Yu YG, Buss GR, Maroof MA (1996) Isolation of a superfamily of candidate disease-resistance genes in soybean based on a conserved nucleotide-binding site. Proc Natl Acad Sci USA 93:11751–11756

    Article  CAS  PubMed Central  PubMed  Google Scholar 

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Authors and Affiliations

  1. Institut Agronomique et Vétérinaire Hassan II, P.O. Box 1123, Agadir, 80 000, Agadir, Morocco

    A. Oumouloud & M. El Otmani

  2. Centro de Investigación y Tecnología Agroalimentaria de Aragón, Apartado 727, 50080, Zaragoza, Spain

    J. M. Álvarez

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  1. A. Oumouloud
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  2. M. El Otmani
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  3. J. M. Álvarez
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Correspondence to A. Oumouloud.

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Oumouloud, A., El Otmani, M. & Álvarez, J.M. Molecular characterization of Fom-1 gene and development of functional markers for molecular breeding of resistance to Fusarium race 2 in melon. Euphytica 205, 491–501 (2015). https://doi.org/10.1007/s10681-015-1420-5

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