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  • 1
    ISSN: 1439-0523
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: A polymerase chain reaction (PCR)-based method was developed to isolate microsatellite markers from large-insert genomic DNA clones of bacterial artificial chromosome (BAC) libraries. The method is fast and economic since it does not require subcloning. It was applied to isolate a microsatellite marker from a BAC clone of the chromosomal region containing the apple scab resistance gene Vf. The Vf gene of Malus floribunda 821 is the most widely used source of scab resistance in apple breeding. A second microsatellite was found on the extremity of a BAC clone flanking the Vf locus. The two microsatellites allowed the identification of the presence of the Vf gene in the scab-resistant accessions M. micromalus SA573-3, ‘Golden Gem’, M. prunifolia 19651 and MA 16 not previously known to carry this gene. They were also used to verify the correctness of the published genealogical tree of the Vf cultivar ‘Florina’, in which a probable mistake was identified. This analysis shows the importance of genotyping the Vf locus when choosing scab-resistant germplasm as parents in breeding programmes.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1617-4623
    Keywords: Key wordsVf ; Malus×domestica ; Resistance gene ; Chromosome walking ; Inverse PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A positional cloning project was started in apple with the aim of isolating the Vf resistance gene of Malus floribunda 821. Vf confers resistance against apple scab, the most important disease in apple orchards. A chromosome walk starting from two molecular markers (M18-CAPS and AM19-SCAR) flanking Vf was performed, using a bacterial artificial chromosome (BAC) library containing inserts of the cultivar Florina, which is heterozygous for Vf. Thirteen BAC clones spanning the region between the two markers were identified in nine chromosome walking steps. The size of the resulting contig is approximately 550 kb. In order to map the Vf region in more detail, we analyzed over 2000 plants from different populations segregating for Vf with markers produced from BAC end sequences. In this way, we were able to restrict the possible location of the Vf gene to a minimum of five clones spanning an interval of approximately 350 kb.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 99 (1999), S. 1012-1017 
    ISSN: 1432-2242
    Keywords: Key words Malus ×domestica ; Venturia inaequalis ; Malus floribunda ; Disease resistance ; Genetic map
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A map-based cloning scheme is being used to isolate the Vf resistance gene of apple against the fungus Venturia inaequalis. Vf is a major dominant gene that is inherited in a Mendelian manner and influenced by minor genes that modify its activity. The two recently published markers M18 and AL07, bracketing Vf, were tested on 1179 progeny plants of three crosses to fine-map Vf. M18 and AL07 were positioned at 0.2 cM and 1.1 cM from Vf respectively, for a total distance between the two markers of 1.3 cM. Physical mapping by pulsed-field gel electrophoresis, using M18 and AL07 as probes, demonstrated that both markers hybridize to a common 870 kb NotI restriction fragment. We therefore found a relationship between physical and genetic distance of 670 kb/cM in the Vf region. This led us to the conclusion that a chromosome walk using the recently published apple BAC library starting from M18 and AL07 is feasible.
    Type of Medium: Electronic Resource
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