Genetic diversity of arabica coffee (Coffea arabica L.) using 20 microsatellite markers in the germplasm bank of UNESUM, Ecuador.
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Abstract
With the objective of determining the genetic diversity of arabica coffee (Coffea arabica L.) by applying 20 microsatellite markers in 20 accessions of the germplasm bank of the Universidad Estatal del Sur de Manabí (UNESUM), young leaflets were collected from the upper middle third of each plant of the 20 coffee accessions conserved in vivo at Finca Andil, in small sealed envelopes and placed in a box with 500 g of silica gel for drying and preservation of the leaflet samples, which were then sent to the Molecular Biology laboratory of the Santa Catalina Experimental Station of the National Institute of Agricultural and Forestry Research (INIAF). The QTA - genotyping analysis was performed with M13 Tailing technology for 20 microsatellite markers. The gels were analyzed visually, determining the weight of the fragments in base pairs (bp) amplified for QTA-genotyping in reference to a marker of known fragment weight. The bp of each marker for each of the coffee accessions (absence/presence) were recorded in an Excel spreadsheet for subsequent statistical analysis. For each marker assayed, a Chi-square test was applied to compare the means of presence levels in the accessions belonging to each marker class (absence/presence) determined, with respect to the bp of the alleles reported by other researchers. The results showed that the SSR markers used have a low or limited detection of genetic diversity, and that limitations could be observed in the levels of heterozygosity and homozygosity at specific loci due to the inability of these SSRs to distinguish alleles from homologous chromosomes, as well as the probability of finding null alleles in polyploids. Moreover, the Cam22 marker was found to be monomorphic.
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