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

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 fu...

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Bibliographic Details
Published in:Euphytica 2015-09, Vol.205 (2), p.491-501
Main Authors: Oumouloud, A, El Otmani, M, Álvarez, J. M
Format: Article
Language:English
Subjects:
amino acid substitution
amino acids
Analysis
Biomarkers
Biomedical and Life Sciences
Biotechnology
complementary DNA
exons
Fruits
Fusarium oxysporum
Fusarium wilt
Genes
Genetic aspects
genetic markers
Genetic polymorphisms
Genetic research
genotype
introns
Life Sciences
loci
marker-assisted selection
Melonis
melons
Molecular biology
pathogens
plant breeding
Plant diseases
Plant Genetics and Genomics
Plant Pathology
Plant Physiology
Plant Sciences
races
Resistance to control
reverse transcriptase polymerase chain reaction
screening
sequence analysis
single nucleotide polymorphism
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Summary: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.
ISSN:0014-2336
1573-5060
DOI:10.1007/s10681-015-1420-5