Os efeitos da luz no metaboloma da cultura in vitro do micélio de Lasiodiplodia Theobromae

Abstract

Fungi defined a group of microorganisms of great industrial and scientific interest. Some strains of species of fungi are used in the production of food, pigments, drinks and medicines. Lasiodiplodia is a genus of the Botryosphaeriaceae family, and a class of Dothideomycetes. This family contains species that occur as saprophytes, parasites or endophytes in a wide variety of host plants, being considered a cosmopolitan species. In order to identify and monitor the metabolites present in the culture of the endophytic fungus Lasiodiplodia theobromae, the fungus was grown in PD medium (Potato-Dextrose) for four weeks, under continuous observation (160 rpm), with the presence and absence of light radiation. Mycelial cultures were collected every seven days, lyophilized and macerated in a buffer solution (sodium phosphate) pH 6.0 and with 10% D2O. As buffer solutions containing extracted metabolites, they were transferred to 5mm NMR tubes and analyzed by 1H-NMR (Bruker 400 MHz). The data obtained were pre-processed using the TopSpin® software, and the region containing the water signal was suppressed using the NOESY1D experiment (pulse sequence noesypr1d). To identify the metabolites, the literature databases were used. The fungus morphologically identified as L. theobromae, had the metabolites isoleucine, leucine, valine, ethanol, lactate, alanine, lysine, acetate, N6-acetylisine, N-Acetylornithine, methionine, acetone, glutamate, succinate, pyruvate, glutamine, aspartate, methylguanidine, betaine, methanol, guanidoacetate, glucose-6-phosphate, glucose, UDP-glucose, uracil, guanosine, fumarate, tyrosine and histidine emerged from L. theobromae cultures grown under both conditions of presence and presence of luminous. The data matrix with the appropriate and normalized values was obtained with the AMIX® software and the multivariate analyzes were performed with the SIMCA® software and an online platform www.metaboanalyst.ca. A multivariate statistical analysis revealed that despite the difference in pigment and biomass production, the metabolism of cultures is qualitatively similar, but quantitatively different, with the highest concentration of the metabolites glycerol, choline, phosphocholine, betaine, acetoacetate, acetone and β-hydroxybutyrate in cultures exposed to light, and a higher concentration of the metabolites tyrosine, phenylacetate, valine, leucine and isoleucine in cultures not exposed to light, revealing that as an exported culture, the photoperiod had its oxidative metabolism intensified, ultimately revealed a greater induced tyrosine oxidation by light, suggesting that L. theobromae cultures perform DOPA-melanin instead of DHN-melanin.