Browsing by Author "Farez Toapanta, Erick Dennis"

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    “Identificación morfológica y molecular de la cepa nativa (Beauveria SPP) 2023-2024”
    (Ecuador : Latacunga : Universidad Técnica de Cotopaxi (UTC), 2024-08) Farez Toapanta, Erick Dennis; Chancusig Espín, Edwin Marcelo
    The objective of this research work was to morphologically and molecularly identify a native strain of Beauveria spp, from the Microbiology Laboratory of the Technical University of Cotopaxi. To reactivate the strains, multiplication methods such as scalpel, toothpicks and seeding loop were used. It was determined that the scalpel turned out to be the most efficient to obtain the isolation of Beauveria spp, since they showed greater sterility and managed to inhibit the contamination of the Petri dishes by bacteria. The macroscopic characteristics of Beauveria spp. Thus, it was possible to identify various morphological structures including hyphae, conidiophores and conidia. Subsequently, the manual extraction of DNA from the fungus was carried out, optimizing a protocol obtained from bibliographic sources and adapting it to the conditions found in the Microbiology laboratory of the Agronomy major. The protocol for PCR (Polymerase Chain Reaction) amplification of the ITS (Internal Transcribed Spacer) region of the fungal genome was standardized and the size of the fragment was verified by horizontal electrophoresis. Next, the amplified samples were sequenced using Sanger sequencing, through the IDGEN Company. Finally, the bioinformatic analysis of the sequences obtained was carried out using the Geneious Prime v2024.0.7 software, and the comparison of the consensus sequence against those reported in the NCBI (National Center for Biotechnology Information) was carried out through the BLASTn application (Basic local alignment search tool). Through molecular identification, it was determined that the fungus corresponded to Trichoderma hamatum. The bases of the protocols for manual extraction of fungal DNA, PCR amplification of the ITS region and bioinformatic analysis were established. It is necessary to carry out the molecular identification of fungi in the laboratory since this way we can know their genus and specific species for which it will contribute significantly to the characterization of microorganisms for the effective application as biological controllers of agricultural pests, thus promoting sustainable and friendly practices. With the environment in the management of crops and ecosystems.