Examinando por Autor "Medina Bardales, Anderson E."
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Ítem De novo assembly and functional annotation of Myrciaria dubia fruit transcriptome reveals multiple metabolic pathways for L-ascorbic acid biosynthesis(BioMed Central, 2015-11-24) Castro Gómez, Juan Carlos; Maddox, J. Dylan; Cobos Ruiz, Marianela; Requena Anicama, David Ruben; Zimic Peralta, Mirko Juan; Bombarely Gomez, Aureliano; Imán Correa, Sixto Alfredo; Cerdeira Gutiérrez, Luis Alexander; Medina Bardales, Anderson E.Myrciaria dubia is an Amazonian fruit shrub that produces numerous bioactive phytochemicals, but is best known by its high L-ascorbic acid (AsA) content in fruits. Pronounced variation in AsA content has been observed both within and among individuals, but the genetic factors responsible for this variation are largely unknown. The goals of this research, therefore, were to assemble, characterize, and annotate the fruit transcriptome of M. dubia in order to reconstruct metabolic pathways and determine if multiple pathways contribute to AsA biosynthesis.RESULTS: In total 24,551,882 high-quality sequence reads were de novo assembled into 70,048 unigenes (mean length = 1150 bp, N50 = 1775 bp). Assembled sequences were annotated using BLASTX against public databases such as TAIR, GR-protein, FB, MGI, RGD, ZFIN, SGN, WB, TIGR_CMR, and JCVI-CMR with 75.2 % of unigenes having annotations. Of the three core GO annotation categories, biological processes comprised 53.6 % of the total assigned annotations, whereas cellular components and molecular functions comprised 23.3 and 23.1 %, respectively. Based on the KEGG pathway assignment of the functionally annotated transcripts, five metabolic pathways for AsA biosynthesis were identified: animal-like pathway, myo-inositol pathway, L-gulose pathway, D-mannose/L-galactose pathway, and uronic acid pathway. All transcripts coding enzymes involved in the ascorbate-glutathione cycle were also identified. Finally, we used the assembly to identified 6314 genic microsatellites and 23,481 high quality SNPs. CONCLUSIONS: This study describes the first next-generation sequencing effort and transcriptome annotation of a non-model Amazonian plant that is relevant for AsA production and other bioactive phytochemicals. Genes encoding key enzymes were successfully identified and metabolic pathways involved in biosynthesis of AsA, anthocyanins, and other metabolic pathways have been reconstructed. The identification of these genes and pathways is in agreement with the empirically observed capability of M. dubia to synthesize and accumulate AsA and other important molecules, and adds to our current knowledge of the molecular biology and biochemistry of their production in plants. By providing insights into the mechanisms underpinning these metabolic processes, these results can be used to direct efforts to genetically manipulate this organism in order to enhance the production of these bioactive phytochemicals.The accumulation of AsA precursor and discovery of genes associated with their biosynthesis and metabolism in M. dubia is intriguing and worthy of further investigation. The sequences and pathways produced here present the genetic framework required for further studies. Quantitative transcriptomics in concert with studies of the genome, proteome, and metabolome under conditions that stimulate production and accumulation of AsA and their precursors are needed to provide a more comprehensive view of how these pathways for AsA metabolism are regulated and linked in this species.Ítem Semillas y plántulas de Myrciaria dubia “camu-camu”: biometría, germinación y crecimiento inicial(Universidad Nacional de Trujillo. Facultad de Ciencias Agropecuarias, 2014-07-05) Medina Bardales, Anderson E.; Córdova Pisco, Evelyn; Fasabi Flores, Anthony; Rodríguez Mashacuri, Napoleón; Cobos Ruiz, Marianela; Imán Correa, Sixto Alfredo; Castro Gómez, Juan Carlos[ES] Los objetivos planteados fueron determinar la biometría de las semillas, registrar la respiración celular en el proceso de imbibición, evaluar el efecto inductor de la germinación de compuestos químicos y analizar el crecimiento inicial. La respiración celular durante la imbibición se midió por reducción del azul de metileno y del Janus Green B. El efecto inductor de la germinación de cinco compuestos químicos fue evaluado. El crecimiento inicial se registró con estereoscopio. Los resultados mostraron que las semillas tienen en promedio un peso de 520±172 mg, largo de 13,50±1,76 mm, ancho de 10,61±1,14 mm y espesor de 4,68±0,62 mm. Estas variables mostraron correlaciones significativas (r de 0,494 a 0,897 y p < 0,001). Las semillas fueron metabólicamente activas durante la imbibición, el cual se evidenció por la reducción de ambos aceptores electrónicos. El nitrito de sodio y el cianuro de potasio indujeron la germinación de las semillas en >90% y 65% respectivamente. El crecimiento hasta plántula constó de seis etapas y ocurrió en un periodo de 22 días después del brote de la radícula. En conclusión, las semillas de M. dubia muestran una amplia variación en sus datos biométricos (peso, largo, ancho y espesor), pero muestran correlaciones significativas entre sí. Asimismo, las semillas son metabólicamente activas en el proceso de imbibición y su germinación es inducida por el nitrito de sodio y el cianuro de potasio. Además, el crecimiento inicial hasta plántula ocurre en un periodo de 22 días después del brote de la radícula y consta de seis etapas bien diferenciadas.--- [EN] The objectives were to determine seed biometrics, recording cellular respiration in the imbibition process, evaluate the effect of inducing chemicals in germination and early growth analysis. Cellular respiration during imbibition was measured by reduction of methylene blue and Janus Green B. The inductor effect of five chemical compounds in germination was evaluated. Initial growth was recorded by stereoscopic. Results showed that the seeds have an average weight of 520 ± 172 mg, length of 13.50 ± 1.76 mm, width of 10.61 ± 1.14 mm, and thickness of 4.68 ± 0 62 mm. These variables showed significant correlations (r of 0.494 to 0.897 and p <0.001). Seeds were metabolically active during imbibition, which was evidenced by reduction of both electronic acceptors. Sodium nitrite and potassium cyanide induced germination of seeds in >90% and 65% respectively. Seedling initial growth consisted of six stages and occurred over a period of 22 days after radicle emergence. In conclusion, M. dubia seeds show wide variation in their biometrics (weight, length, width and thickness), but show significant correlations with each other. Likewise, seeds are metabolically active in imbibition process, and germination is induced by sodium nitrite and potassium cyanide. Furthermore, seedling initial growth to occur over a period of 22 days after emergence of the radicle and consists of six distinct stages.