Nuevos progresos en el proyecto de investigación…

En lo que va de semestre, hemos estado trabajando fuertemente para cumplir a cabalidad las metas propuestas para este semestre. Hasta el momento, hemos estado purificando cepas, congelándolas una vez estas se encuentran puras y haciendo extracción de DNA. Para la próxima semana vamos a estar haciendo PCR con el propósito de amplificar el 16S RNA y luego hacer electroforesis de gel.

Cabe destacar, que hemos tenido algunas dificultades en estos días, debido a que se nos acabaron algunos materiales, como por ejemplo los platos Petri, y hasta que no llegue la orden que enviamos a comprar, no podemos continuar con la purificación de cepas, ni pasando las cepas a platos nuevos para su crecimiento. Debido a esto hemos decidido continuar con las extracciones de las cepas que tenemos congeladas, y realizar PCR. Estamos alternando algunos pasos en nuestra investigación, con el propósito de continuar trabajando efectivamente, y no tener que parar debido a la falta de ciertos materiales.

En cuanto al progreso de la investigación en una escala del 1-5, con relación a las metas establecidas en el mes de enero, puedo clasificarla como un 2. Entiéndase que llevamos algo de progreso, logrando al menos un 10% de los objetivos propuestos. A medida que continúe el semestre esperamos poder cumplir con todos los objetivos propuestos, o por lo menos con la mayoría de ellos.

Abstract de mi proyecto de investigación:

Extremophilic Diversity in Mangrove’s Rhizosphere; a Source of Putative
Novel Microorganisms with Biotechnological Potential

Mangroves are ecosystems of great importance for the natural maintenance of the coastal lands by complex interactions. The presence of halophilic bacteria had been documented on the leaves of these plants, but little is known about the microbial diversity present on the black mangrove rhizosphere. This is the reason why this research is oriented towards the determination of the microbial diversity present at these mangroves. An analysis of the diversity of halophilic or halotolerant microorganisms present in the soil of these black mangroves rhizosphere found in two different localities in the south area of Puerto Rico, the Salinas of Cabo Rojo and the Boquerón National Forest was carried out using culture-dependent and culture-independent techniques. Soil samples from six different plants of each locality and at three different depths (0cm, 10cm and 20cm) were taken and processed. As a culture-dependent, Seghal Gibbon’s (SG) media was used and subsequently techniques of purification in four quadrants and micro dilutions were applied. By using the culture-dependent technique, a representative sample of the diversity present in the soil was not recovered, so a DNA total extraction and PCR of 16sRNA were used to determine the presence of bacteria. Techniques like Terminal Restriction Fragment Polymorphism (TRFLP), was used to determine the diversity of the soil; and environmental gene libraries, which are used to obtain sequences of possible microorganisms that live in those places and are not cultivable, is going to be applied. This project hypothesizes whether there are halophilic/halotolerant microorganisms present in the black mangroves and if so, does this microbial diversity vary according to the depth and/or place. Thus far, it has been found that the black mangroves in the Salinas and in Boquerón contained presence of halophilic and halotolerant microorganisms. These characteristics are determined by physiological characterizations. Also, from the 16sRNA samples from two different plants at 0cm and 20cm from each location were amplified successfully. From the PCR of the TRFLP, without digestion, amplification of all the Boquerón samples was obtained. The results obtained from this research will provide information about the physiology and chemistry of the microbial populations interacting with the plant and provide novel organisms with biotechnological potential as well.

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