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Molecular Characterization of Wild Microorganisms Associated with Artisanal Cacao Fermentations in the Municipality of Los Andes, Nariño, Colombia
This study focuses on the molecular characterization of microorganisms associated with cacao fermentation in the department of Nariño, Colombia. Through the analysis of the ITS1-5.8S-ITS2 and D1/D2 rRNA (26S) regions for yeasts, and 16S rRNA for lactic acid bacteria (LAB) and acetic acid bacteria (AAB), various microorganisms involved in cacao fermentation were identified. Additionally, the in-silico RFLP (Restriction Fragment Length Polymorphism) technique was applied to resolve intraspecific relationships and establish differentiated molecular patterns. Isolates of Saccharomyces cerevisiae, Pichia kudriavzevii, Candida tropicalis, Candida glabrata, Levilactobacillus brevis, Lactiplantibacillus plantarum, Acetobacter fabarum, Acetobacter okinawensis, and Acetobacter tropicalis were identified. The use of restriction enzymes such as AluI, MseI, HinfI, HhaI, and HaeIII was key to discriminating between species and resolving intraspecific relationships in the clades. These results provide a solid foundation for better characterizing the microbial diversity in cacao fermentation in this region.
