AFLP and RAPD Characterization of Phaeoacremonium aleophilum Associated with Vitis vinifera Decline in Spain |
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Authors: | Laura Martín Luis E Sáenz de Miera María Teresa Martín |
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Institution: | 1. Instituto Tecnológico Agrario de Castilla y León (ITACYL), , 47071 Valladolid, Spain;2. área de Genética, Universidad de León, , 24071 León, Spain |
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Abstract: | The genetic diversity among Spanish isolates of the fungus Phaeoacremonium aleophilum, one of the major causes of grapevine decline, was determined using random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) techniques. Using RAPD, a large genetic variation was observed among 36 Pm. aleophilum single‐spore cultures, with 76 (82.6%) polymorphic bands generated by 12 RAPD primers. A neighbour‐joining dendrogram showing the RAPD patterns of diversity revealed four groups of haplotypes. The Bayesian and principal components clustering analysis revealed three groups of haplotypes. When more than one isolate of Pm. aleophilum was obtained from a single vine, different haplotypes were found. Seventeen single‐spore isolates were used for AFLP analysis. Five primer combinations produced 358 scorable markers, of which 309 (86.3%) were polymorphic. The analysis based on genetic distance as well as clustering analysis confirmed three main groups largely in agreement with those returned by the RAPD results. The Mantel correlation between the RAPD and AFLP distance matrices ranged from R = 0.5931 to R = 0.6294. The high level of haplotype diversity among the RAPD and AFLP markers suggests that sexual reproduction and genetic recombination may occur between Pm. aleophilum haplotypes in Spain. The AFLP approach revealed a greater number of polymorphic markers. A relationship between the genetic profile of the infecting isolate of Pm. aleophilum and the age or decline symptoms of the grapevines may exist. |
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Keywords: | esca and Petri disease Eutypa dieback genetic diversity grapevine molecular markers |
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