| Non-Scientific
Abstract Phytoplankton are one-celled organisms
that live in either fresh or salt water and perform photosynthesis. They
are responsible for about 40% of the planetıs total annual
photosynthetic production.However, it is very difficult to measure the
number of species of phytoplankton and how many of each kind there are
in a sample of seawater. In the past, phytoplankton diversity assessment
has been done by looking at physical characteristics, but microorganisms
are often very small and very similar looking.Particular gene sequences,
however, are usually unique to species. The goal of this study was to
develop a method, using new highly sensitive and efficient molecular
biology techniques, to assess the genetic diversity in complex marine
eukaryotic microbial communities and to assess the protocolıs efficacy.
Such a method would be important for measuring microbial community
structure across naturally occurring and artificially generated
differences in the ocean.A method was developed to get the DNA from the
cells, make many copies of the DNA with a label on one end, and cut the
DNA.Using T-RFLP (terminal restriction fragment length polymorphism), a
machine can plot how long each fragment of DNA is (just the piece that
is labeled) by how many there are. Theoretically, each fragment length
should be unique to each species.All steps were completed with exception
of T-RFLP, which is still in progress. Each step was developed
successfully except for the amplification of the DNA.
Abstract There is a growing debate among scientists
whether iron fertilization in the oceans will lead to more carbon
fixation by the oceanıs phytoplankton. As a result, it is more important
than ever to develop a quick, easy, and accurate way to assess
phytoplankton diversity and community structure. Historically, marine
microbial organisms have been characterized morphologically though it is
difficult to differentiate some groups using morphological criteria
because of their small size and lack of distinctive taxonomic
characteristics. The goal of this study was to develop a protocol, using
new highly sensitive and efficient molecular biology techniques, to
assess the genetic diversity in complex marine eukaryotic microbial
communities and to assess the protocolıs efficacy. Given their extreme
diversity, eukaryotic phytoplankton was used as a model community to
study microbial marine genetic diversity. The resulting protocol
includes: cell filtration,DNA extraction, PCR amplification, restriction
digests, and T-RFLP.All steps were completed with exception of T-RFLP,
which is still in progress. Successful DNA extraction and restriction
digest methods were found that worked for all species tested, though
there were some problems with PCR amplification, including preferential
amplification and non-specific PCR products. |
