Banana is one of the most important edible crops in the world, however, it’s attacked by different pathogens, one of the most prominent of these is the fungus Mycosphaerella fijiensis, causal agent of Black Sigatoka. The environmental and economic issues related to the pesticides used for its control have encouraged the search for cleaner alternative biomolecules. Previous studies were made by the Biotechnology group from the Universidad de Antioquia searching for a biological alternative for the control of M. fijiensis; in these the antifungal capacity of the fungus Ganoderma lucidum was determined as an antagonist and then as a source of protein extracts with inhibitory activity in vitro and in greenhouse plants; these findings were the foundation of this work, which focuses in the study of the enzymatic capacity of the proteins present in the protein extracts, due to their potential ability to degrade different compounds, including polysaccharides, lipids, peptides and nucleic acids, constituents of essential parts of a living cell; therefore these extracts can act as possible antifungal agents. In this study, protein extracts of G. lucidum obtained from bioreactor cultures (BIOFLO 110 ®) were characterized in terms of their deoxyribonuclease, ribonuclease, protease, glucanase and chitinase enzymatic activities. The extracts were also fractionated and each fraction obtained was evaluated for its inhibitory capacity against the phytopathogen fungus M. fijiensis, and through mass spectrometry analysis the presence of different enzymes with antifungal potential was confirmed.
<p class="p1"><strong>ABSTRACT</strong></p><p class="p2"><em>Stevia rebaudiana</em> (Asteraceae) is a plant of economic importance because of its medicinal properties and the presence of sweetener compounds on its leaves. These compounds can be a substitute for sucrose in a wide variety of products used by persons with diabetes and obesity problems. To standardize an efficient and effective propagation method for the different Stevia genotypes grown in Colombia, this study evaluated the effect of different combinations of the plant growth regulators 2,4-dichlorophenoxyacetic acid (2,4-D), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), 6-(gamma, gamma-dimethylallylamino) purine (2iP) and Zeatin on the induction and development of somatic embryos. Adenine and coconut water were also evaluated as supplements in the basal culture medium Murashige and Skoog Basal Salt Mixture (MS) with glutamine. The combination of 2,4-D (18.09 μM) and 2iP (7.38 μM) produced the highest number of somatic embryos per explant, which had well-defined characteristics. The genotype showed a significant effect on the embryogenic response. In the “SRQ-93” genotype, the formation and development of somatic embryos was achieved, whereas the genotypes “Bertoni” and “Morita II” only yielded embryogenic and non-embryogenic calli, respectively. The conversion to seedlings was achieved on the regeneration medium containing gibberellic acid (GA<span class="s1">3</span>) (0.29 μM) and activated charcoal. </p><p class="p1"><strong>RESUMEN</strong></p><p class="p2"><em>Stevia rebaudiana</em> (Asteraceae), es una planta de gran importancia económica debido a sus propiedades medicinales y a la presencia de compuestos endulzantes en sus hojas, los cuales pueden sustituir la sacarosa en gran variedad de productos utilizados por personas con problemas de diabetes y obesidad. Con el propósito de estandarizar un método de propagación eficiente y efectivo para diferentes genotipos de Stevia cultivados en Colombia, en la presente investigación se evaluó el efecto sobre la inducción y desarrollo de embriones somáticos de diferentes combinaciones de los reguladores de crecimiento vegetal 2,4-D, IAA, IBA, 2iP y Zeatina, además de los suplementos adenina y agua de coco en el medio de cultivo basal Murashige y Skoog (1962), adicionado con glutamina. Con la combinación 2,4-D (18.09 μM) y 2iP (7.38 μM) se obtuvo el mayor número embriones somáticos por explante con características bien definidas. El genotipo tuvo un efecto significativo en la repuesta embriogénica, en el genotipo “SRQ-93” se logró la formación y el desarrollo de embriones somáticos, mientras que en los genotipos “Bertoni” y “Morita II”, solo se obtuvo callo embriogénico y no embriogénico respectivamente. La conversión a plántulas se alcanzó en el medio de regeneración conteniendo GA3 (0.29 μM) y carbón activado.</p><p class="p2"> </p>
In order to understand the causes of lack of regeneration in cacao somatic embryos, two cacao varieties with different responses to regeneration potential were described based on their capacity to store different compounds. It is well known that seed reserves play a central role in the regenerative capability of somatic embryos; thus, we followed histochemical changes and reserve fluctuations of proteins, polysaccharides and polyphenols during somatic embryogenesis (SE) in the two cacao varieties. The study showed that, in somatic embryos of the regenerating variety, polyphenols were localized mainly in the periphery of the embryo (epidermal cells) and proteins were the main storage substance in the embryo expression medium, while the non-regenerating variety had a high presence of polysaccharides with random distribution of polyphenols at the end of the embryo induction step.Keywords: antioxidants, histology, reserve accumulation, recalcitrance, somatic embryogenesis. RESUMENDos variedades de cacao con diferentes respuestas a la regeneración fueron descritas en función de su capacidad para almacenar diferentes compuestos, con el fin de aproximarse al entendimiento de las causas de la falta de regeneración en embriones somáticos de cacao. Es bien sabido que las reservas de semillas desempeñan un papel central en la capacidad de regeneración de embriones somáticos; por tanto, se realizó un seguimiento de cambios histoquímicos y fluctuaciones de reserva de proteínas, polisacáridos y polifenoles durante la embriogénesis somática (SE) en dos variedades de cacao. El estudio mostró que, en los embriones somáticos de la variedad regenerante, los polifenoles se localizaron principalmente en la periferia del embrión (células de la epidermis) y las proteínas fueron el componente principal de almacenamiento en el medio de expresión de embriones, mientras que la variedad no regenerante tenía una alta presencia de polisacáridos y una distribución aleatoria de los polifenoles en el final de la etapa de inducción de embriones. Palabras clave: acumulación de reservas, antioxidantes, embriogénesis somática, histología, recalcitrancia.
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