Evaluación de comunidades microbianas edáficas oxidantes del amonio durante un ciclo de cultivo de papa criolla (Solanum phureja)
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La nitrificación depende enteramente del metabolismo microbiano e inicia con la transformación del amonio a hidroxilamina, catalizada por la enzima AMO (Amonio Monooxigenasa). El avance de las tecnologías independientes de cultivo, ha permitido conocer la secuencia del gen que codifica para el sitio activo de dicha enzima (amoA), identificándolo como un potente marcador taxonómico y funcional especifico para el rastreo de las comunidades microbianas oxidantes del amonio (COA) en diversos ambientes. El objetivo de este trabajo fue evaluar las COA edáficas asociadas a un cultivo de papa (Solanum phureja) ubicado en la sabana de Bogotá D.C, con un manejo orgánico de 20 años, a través de la cuantificación de la abundancia del gen amoA por qPCR usando los cebadores específicos. Los valores de abundancia no pudieron ser determinados por problemas inherentes a los cebadores utilizados para la qPCR, que pese a ser ampliamente implementados para este tipo de estudios en diferentes ambientes, no cumplen los criterios requeridos por la técnica de cuantificación Sybr green, causando la formación de dímeros de primers de alto Tm, que interfirieron con la eficiencia de la amplificación, demostrando que no son adecuados para la determinación de la abundancia del gen amoA de los suelos evaluados.
Abstract. Nitrification depends entirely on microbial metabolism and begins with the conversion of ammonia to hydroxylamine, catalyzed by AMO (Ammonia Monooxygenase). The progress of independent culture technologies has allowed the knowledge of enzyme active site of the (amoA) gene and identified it as a powerful taxonomic and functional marker specific for tracking microbial communities of ammonia oxidizers (COA) in different environments. The aim of this study was to evaluate the associated soil COA potato crop (Solanum phureja) located in the savannah of Bogotá, with organic management 20 years, through the quantification of amoA gene abundance by qPCR using specific primers. Abundance values, could not be determined because of inherent problems in the primers used for qPCR, which despite being widely deployed for such studies in different environments, do not meet the criteria required by the quantification technique, causing the formation of primers dimer that interfered with the efficiency of the amplification, demonstrating that they are not suitable for determining the abundance of the gene amoA of soils evaluated.
Abstract. Nitrification depends entirely on microbial metabolism and begins with the conversion of ammonia to hydroxylamine, catalyzed by AMO (Ammonia Monooxygenase). The progress of independent culture technologies has allowed the knowledge of enzyme active site of the (amoA) gene and identified it as a powerful taxonomic and functional marker specific for tracking microbial communities of ammonia oxidizers (COA) in different environments. The aim of this study was to evaluate the associated soil COA potato crop (Solanum phureja) located in the savannah of Bogotá, with organic management 20 years, through the quantification of amoA gene abundance by qPCR using specific primers. Abundance values, could not be determined because of inherent problems in the primers used for qPCR, which despite being widely deployed for such studies in different environments, do not meet the criteria required by the quantification technique, causing the formation of primers dimer that interfered with the efficiency of the amplification, demonstrating that they are not suitable for determining the abundance of the gene amoA of soils evaluated.