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One of the techniques I learned during my classes and laboratories is the extraction and amplification of DNA. I will use this technique in my project to make a more specific study of the types of fungi that Thalassia testudinum have encrypted inside the leaves. This experiment will be conducted with normal leaves and leaves after processing them with the rigorous sterilization protocol that is used to cultivate them in vitro. After extracting the DNA it will be sequenced to ensure that we have in the “endophytic fungi library” all the cultivable species that are present inside T. testudinum leaves.

Using a scale of 1-5; I would say that my project has only a 2 because wrong materials came to the laboratory and we had to wait several weeks for the correct. But we already have the aquarium system with T. testudinum growing , one of our goals from the beginning.

Aspergillus sp.

Here we go again!

What’s new?!?!

Hello!!!! Welcome to my blog!

For this semester our laboratory team has the same objectives of last semester, but we are trying new ways to reach our goal. Our most important task is to obtain axenic cultures of Thalassia testudinum to study mutual relationships between the endophytes and the plant.

At this time, we are collecting samples; on Buye Beach in Cabo Rojo, Puerto Rico, looking for flowers, seeds and plants of T. testudinum. Last semester we were doing in vitro cultures of tissue from the rhizomes, but for this semester will use tissue from flowers, leaves, rhizomes and seeds. We will do these various tests to see which tissue is best developed and which has less population of microorganisms.  Also, we will be changing the pH and concentration of growth factors in culture media. All these arrangements are being made to optimize the growth conditions of axenic T. testudinum.

During this semester I will develop several skills like culturing tissue at a cellular level, working with new protocols for sterilization, preparation of media for different types of tissue, and work in microscopy using scanning and transmission electron microscopes.

Thalassia testudinum figure

Visiting the bio-blogs

It was a very enriching experience to visit the bio-blogs of other young researchers like me. This assignment gave me the opportunity to learn about other research projects that are being carried out in our college system. I really enjoy the experience because the projects themes where very different, but in the lines written by my companions I found something in common, the passion of someone who enjoys what is doing, and the great interest that we share for research and problem solving.

Coqui

The first bio-blog I visited was Josue Negron’s blog. He is an UPRH biology student and is working with new amphibian species. Right now his team is documenting the basic reproductive parameters of a recently discovered Coqui, Eleutherodactylus juanariveroi. Also they are orienting the community about these new species, and I believe this is very important so they have the knowledge that is a need to preserve this species.

The second bio-blog I visited was Odemaris Narvaez’s blog. She is an UPRM industrial microbiology student and is working with algae and the different organisms that live in association with them. Currently she is working with a cyanobacteria called Microcystis aeruginosa. This organism can cause great damage to the tissue it infects, for example the toxin its produces can cause liver cancer. I found her research very interesting because she is going to isolate the DNA that codifies for the microcystin (toxin) of M. aeruginosa and make different tests in human tissues to observe their interactions.

The third bio-blog I visited was Keren Valentin’s blog. She is an UPRM chemical engineering student and is working with electrochemistry using a technique called cyclic voltametry to study the electroactivity of Hemoglobin I (HB I). It is important to study this molecule because it has a mechanism involved in the transportation of H2S, also HB I has great affinity for this compound; this characteristics give an excellent idea to develop a biosensor capable of identify and quantify H2S directly. The development of this biosensor is very important because the ones that exist in these days are not so good. H2S is related with contamination (like the presence of certain bacteria) and several diseases.

Last post, November 2009!

One of the most important lessons that I have learned this semester is to be patient and perseverant. In vitro plant tissue culture is a process that has to be carried in extremely sterile conditions and the person working with the tissue (me) has to be very careful to avoid tissue damage. Also I have to wait long time in order to obtain positive results (tissue growth and no contamination).

At this time my second trial on in vitro cultures of Thalassia testudinum is growing very well! I am making new nutritive medium with different concentrations of auxines and cytokinins (auxines are hormones that affect plant growth, they can be characterized by their ability to induce cell elongation in stems, and meanwhile cytokinins are compounds which promote cell division), to transplant the tissues with the best development range and determine the best growth conditions for the T. testudinum tissues.

My next semester is full of hard work from beginning to end! Once we have all the tissue growing in the required conditions, a system of little aquariums would be installed and part of the plants will be put in there, in order to induce propagation of the sterile clones, and the other part will be use in different tests with endophytic fungi to determine their function in the plant.

Second Post: October, 2009

Hello! Welcome to my blog!

I hope this month has been full of enriching experiences for you! Every day is a new opportunity given to us to learn new things, so take this privilege to the fullest! As part of this blog entry I will share my experiences during this month in a new research project.

Currently there is a wide range of articles dealing with the subject of micropropagation of plants and seagrasses where they explain clearly what micropropagation means and implies procedures and variations used to improve the development of certain specific plants. But no article has been made on in vitro micropropagation of Thalassia testudinum tissue. In many articles mentioned the great difficulty with this seagrass when it comes to growing in a laboratory under sterile conditions, but has never been effectively micropropagate. So, as I said in the last post, our job is to find an effective method to achieve micropropagation of T. testudinum tissue.

After a month of working in the laboratory on my new project I had the opportunity to interact more with undergraduates and graduates students working under the tutelage of my mentor, Professor Sandra Maldonado-Ramirez. I have had the opportunity to work with other professors and laboratories. Thanks to this experience and Prof. Dimuth Siritunga, I have learned new techniques for tissue culture and new ways to keep these tissue cultures under strict aseptic conditions. Right now I’m making several tests to see under what conditions the tissue culture is better developed. We are ranging from light-dark cycles to concentrations and environmental conditions; there are many variables that we must take into consideration until we find the right conditions for the better development of tissues. Once we get the ideal conditions and begin proliferating normal tissue, we will proceed to the other stage of the investigation.

Welcome!

Hi!

My name is Ileana Marrero-Berríos, and I’m an undergraduate student of the Industrial Biotechnology program at UPR Mayaguez, Colegio!

Currently, I’m working with Prof. Sandra Maldonado-Ramirez in the Mycology and Aerobiology Laboratory. We are conducting a research on endophytic fungi and the microbial nitrogen cycle on in-vitro cultures of the tropical seagrass Talassia testudinum. Endophytic fungi are organisms that live in association with plants in most or all their life cycle. They grow between plant tissues without any outward manifestation. Previous studies have shown that these have an important role in plant tissue they inhabit. One of our objectives is to determine these roles. In order to perform this research, we have to micropropagate T. testudinum; micropropagation is a way to clone plants axenically (sterile). But, till this day no one has successfully propagated T. testudinum in sterile conditions, so we have a great challenge on our hands.

Me!!

I began my research project with Prof. Sandra Maldonado-Ramirez last semester (January-May, 2009).
Since that day, I started working with T. testudinum and endophytes. I was interested in making a survey of endophytic fungi in tissue of T. testudinum and compare it to a survey held by Selimar Malavé-Vega in 2006. We observed the variation in the population of endophytes with the pass of time and climate changes in Buyé Beach in Cabo Rojo, Puerto Rico. I really enjoy this experience, and I learned more about aseptically techniques, literature search and how to write a good scientific report.

Prof. Sandra Maldonado Ramirez PhD

First post- August 27, 2009

Hello!

My name is Ileana Marrero-Berrios. I’m an Industrial Biotechnology student at UPR Mayaguez.