Genotyping of Vanilla planifolia Jacks ex Andrews from the Totonacapan region using Inter-Simple Sequence Repeat markers

Abstract

Vanilla is the only genus of plants in the Orchidaceae family used in the food industry, being the most widely used flavoring worldwide and the second most valuable economically. Cultivation is of great biocultural importance in the Totonacapan region, in the state of Veracruz, as this region is considered its center of origin and distribution. However, little is known regarding the number of genotypes present in this area, as well as the genetic diversity, which is vital for the improvement of this crop. Therefore, the objective of this study was to perform the genotyping of 60 Vanilla planifolia accessions from the Germplasm Bank of The National Institute of Forestry, Agriculture, and Livestock Research (Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias - INIFAP), Experimental Field Ixtacuaco, which were selected and collected in the Totonacapan region. For this purpose, the number of V. planifolia genotypes was determined based on ISSR (Inter-Simple Sequence Repeat) molecular markers. Three DNA extraction protocols from plants and fungi described in the literature were evaluated, and a fourth protocol was adapted with different procedures from the previous protocols, seeking to obtain a high concentration and good quality and integrity. Once quality DNA was obtained, fourteen ISSR primers were evaluated, of which three primers that showed the highest number of amplified bands were selected. Subsequently, the genotyping of the 60 accessions was carried out, resulting in nine genotypes with primer 808 and six genotypes with primers 812 and 856. These results allowed the genotyping of Vanilla planifolia accessions with ISSR markers, which will help evaluate the genetic diversity of this crop in the Totonacapan region in the future.