Desenvolvimento de um método molecular para a identificação de clones de variedades de Coffea canephora

Abstract

Recent advances in coffee genomics have enabled the development of several molecular tools for the identification and characterization of the available germplasm for genetic improvement of coffee, as well as allowed the development of molecular methods for the identification and differentiation of genetic materials. Therefore, the objective of this work was to develop a fast and low-cost molecular method for the identification of Coffea canephora clones. First, we selected SNPs (Single Nucleotide Polymorphisms) that were inserted into the cutting site of EcoRI restriction enzyme and that had allelic discrimination. Primers flanking this region were designed, PCRs were performed, and the amplification products were enzymatically cut with EcoRI. After performing electrophoresis, the resulting bands were compared with those of the control genome, allowing the identification and description of the alleles. Our results show that the developed method was efficient in the genetic discrimination of several coffee clones from varieties present in the National Cultivar Protection System of the Ministry of Agriculture, Livestock and Food Supply (MAPA).