Phytophthora belongs to a group of plant pathogens found in the Kingdom Protista and is known to be highly destructive to plants. The water mold has been found all over the U.S. but just in Washington and Oregon, there are over 30 different species. This water mold is a significant problem in the Pacific Northwest because it infects Pacific Rhododendron leaves. Comprehending the factors that contribute most to the diversification of the Phytophthora species is essential in sourcing and minimizing its spread. We anticipate that the presence of Phytophthora species in residential areas, influenced by human activities, will procure a higher diversity rate of at least 3 or more species. This expectation is rooted in the understanding that the human influence on these environments contribute to an increase in landscaping that works for the proliferation of diverse Phytophthora species. Our location sampling of choice for the bacteria was Chambers Creek located in Pierce County, WA. Chambers Creek has mostly residentially areas surrounding. We gathered four leaves from a rhododendron plant on the Pierce College Fort Steilacoom. We placed the collected rhododendron leaves into bags which were then submerged in the water and left undisturbed for a duration of approximately nine days for observation and analysis. We plan to inoculate the necrotic tissue into V8 agar. We plan to isolate DNA of the Cox-1 subunit protein and sequence it to then use bioinformatics to identify species of Phytophthora. We will use class data to determine if residential areas have an effect on Phytophthora diversity.

Our undergraduate research project is on Phytophthora. Phytophthora translates to “plant destroyer”. The purpose of our research is to use watershed studies and bioinformatics to find patterns in the diversity of Phytophthora species found in local streams in Washington State. We assessed prior studies to estimate the probability of discovering Phytophthora in Chambers Creek, a local stream. Our hypothesis is, in comparison to freshwater, water sources closer to the ocean will have a higher diversity of Phytophthora species because they have more sources of contamination and higher saltwater concentrations. We chose the stream location based on previous watershed information we had. Additionally, we baited Chambers Creek with Rhododendron leaves in an attempt to grow Phytophthora. We then cultured the lesions on the leaves onto a V8 agar to grow Phytophthora. After that we isolated the DNA and “cleaned” it by using reagents to remove unused primers and deoxynucleoside triphosphates, then sent it off for sequencing. At this time, we used gel electrophoresis to determine the quality and quantity of our DNA. Lastly, we used bioinformatics to become familiar with sequence analysis using the National Center for Biotechnology Information Basic Local Alignment Search Tool. Three out of the four samples we obtained had no sequence matches but one sample was 100% positive for Phytophthora bilorbang. In conclusion our hypothesis was correct. Since our water source was subject to higher saltwater concentrations, it was more contaminated and had a higher diversity of Phytophthora species. Doing this research and discovering this information will help add to the knowledge of Phytophthora in Washington State. This can lead to where Phytophthora treatment is needed to protect our environment.