Morphological and Molecular Identification of Mycobiota Associated with Juniperus monosperma (Engelm.) Sarg. Blight in Mexico

Abstract

The aim of the investigation was to identify the agents associated with blight in Juniperus monosperma at Lirios region, Arteaga, Coahuila. Botanical material was collected at Lirios, Arteaga, and taken to the laboratory. Pathogens were isolated in ADP culture medium and identifying by morphological criteria and molecular using primers ITS1 and ITS4. DNA extraction by the Dellaporta method, the visualization of obtained DNA was performed by electrophoresis on a 2% (p/v) agarose gel. DNA quantification was performed on a NanoDrop 1000 spectrophotometer, and DNA amplification was carried out in the Veriti thermocycler. Obtained sequences were aligned and compared with those available in the GenBank database of the National Center for Biotechnology Information (NCBI), using the BLAST algorithm (Basic Local Aligment Search Tool) to find conserved sequences. Pathogens were inoculated in stems of J. monosperma. The agents associated with blight were Alternaria sp., Aspergillus sp. and Rosellinia sp. and the sequence obtained compared with BLAST coincided only with Alternaria sp. and Aspergillus sp. with the access codes KP027305.1 and MG551283.1, respectively. According to data taken in field, Alternaria sp. behaved as the highest prevalence species and severity associated with blight in J. monosperma, and Rosellinia sp. in laboratory conditions.