Pressley Wilson, University of South Carolina Aiken
The Approach: In my previous post, I discussed (1) the importance of an organism’s microbiome in relation to its health and (2) the importance of algae in marine ecosystems due to their ability in producing oxygen, removing nitrogen and phosphorus from water, and exchanging inorganic carbon.
Considering the importance of these two variables, this summer I am researching the relationship between the algae microbiome and algae species in One’ula Beach, Honolulu, Hawai’i.
The objectives of my research project are:
- Identify relationship between algae microbiome and algae species
- Identify relationship between the microbiome with algae’s morphology
This study will predict there is a variation in the microbiome between algae species, due to the different species characteristics, such as calcification.
The five algal species of different morphologies were hand-sampled by Dr. Heather Spalding at the intertidal region of One’ula Beach. The algae samples were rinsed with artificial saltwater to remove dirt and loosely associated bacteria. After cleaning, each species were placed into (1) a micro-centrifuge tube with 0.5 mL of RNA-later or (2) a 15 milliliter conical tube with 1.5 mL RNA-later. The RNA-later is a DNA preservation agent, was used to stabilize the algae DNA. The samples were stored at 4°C overnight. This overnight incubation allowed the RNA-later to penetrate the bacterial and algal cells to the DNA. After this incubation all tubes were frozen at -20°C and shipped overnight on dry ice to the College of Charleston, South Carolina, where they were stored in -80°C freezer until DNA extraction.
Sample Analysis a
A MoBio Fast DNA Spin Kit was used to extract the DNA from the algal samples. This DNA is then tested by PCR to determine if bacteria are present on the algae. To determine the number of bacteria present, qPCR is used. This molecular biology technique is used to quantify specific genes in a sample.
The PCR samples will be analyzed using gel electrophoresis, a molecular biology procedure that uses an electrical current to separate the components of the sample DNA by size. The qPCR data will be compared to a known DNA standard to determine the number of bacteria in our samples and calculations will be performed using excel.
I would like to thank Dr. Heather Fullerton for her guidance and support with this project and Dr. Heather Spalding for her sample collection. This project is supported by the Fort Johnson REU Program, NSF DBI-1757899.