Aquatic Plant Life Biodiversity in Long Island’s Rivers Madison DiPalo, Hanna Engelhardt, & Alison Kovarik Mentor: Mr. James Ostensen Eastport- South Manor Junior Senior High School Discussion Most of the species we identified are non-native but still share vital roles in the local ecosystems of Long Island. Examples include Bur-Reed which is an abundant food source for many local organisms such as, the White Winged Scoter Ducks, the Mute Swan, and other waterfowl. The Eelgrass and the Field Pepperwort provide habitat as a breeding ground for native insects such as the Green Darner dragonfly. Eelgrass is a vital, native species to Long Island’s rivers. Eelgrass is an indicator species that indicates a productive and healthy ecosystem with proper water quality conditions. Eelgrass has seen a decline due to the long term exposure to European Swan species. These swans not only consume the Eelgrass in abundant amounts, they cause eutrophication due to their large production of fecal matter. Where a problem arises is concerning native species of Ulva. Ulva rigida, along with Ulva scandinavia (more commonly known as ‘sea lettuce’) has become a major problem concerning nutrient abundance in rivers. Factors such as heavy reliance on fertilizers, and pesticides on Long Island, along with faulty sewer systems such as those in Mastic- Shirley along the Forge River has allowed excess nutrient leaching, such as excess phosphate, and nitrogen, into the rivers. Ulva thrive in conditions with excess nutrients. Ulva, a form of algae, under the right conditions, cause algal blooms which halt the production of oxygen through photosynthesis among bottom dwelling river plants such as Eelgrass. This decreases the amount of available dissolved oxygen in rivers, which can cause problems such as massive fish die-offs. In extreme cases, as of that in Brittany Beach, France, thousands of pounds of algae washed up on the beach due to the presence of excess nutrients. The decomposition of the algae let off noxious gas that killed off local wild boar. This algae caused many cases of sickness in the area among the death of a truck driver who was attempting to remove the algae. The gas caused the driver to pass out, and crash the truck resulting in fatality. References Figure 2f from: Irimia R, Gottschling M (2016) Taxonomic revision of Rochefortia Sw. (Ehretiaceae, Boraginales). Biodiversity Data Journal 4: E7720. https://doi.org/10.3897/BDJ.4.e7720. (n.d.). doi:10.3897/bdj.4.e7720.figure2f Peconic Estuary – Protecting & Restoring Long Island's Peconic Bays. (n.d.). Retrieved from https://www.peconicestuary.org/ (n.d.). Retrieved from https://www.brookhavenny.gov/Elected-Officials/Council-District-6/Forge-River Zostera marina L. (n.d.). Retrieved from https://gobotany.newenglandwild.org/species/zostera/marina/ Giant Bur-Reed (Sparganium eurycarpum). (n.d.). Retrieved from http://www.illinoiswildflowers.info/wetland/plants/gnt_burreed.html Lepidium campestre (L.) Ait. f. (n.d.). Retrieved from https://gobotany.newenglandwild.org/species/lepidium/campestre/ Acknowledgements A thank you to Stony Brook University for letting us use your PCR extraction equipment, and being so incredibly helpful. Also, a thank you to Cold Spring Harbor Laboratory for assisting us in sequencing of our samples. Lastly, a thank you to Barcode Long Island for conducting and establishing this project. Our teacher mentor Mr. Ostensen and Eastport South Manor High School. Abstract Biodiversity is crucial to maintaining ecosystem stability. Measuring the volume of different organisms and their functions in the environment can provide us with a greater understanding of the health and well-being of the environment. Biodiversity is used as an indicator of the well-being of an environment. The presence of luscious plant life in an ecosystem indicates a healthy ecosystem, while the presence of algal species can indicate an unhealthy ecosystem experiencing problems such as eutrophication, low dissolved oxygen, along with many other issues. These issues, along with a lack of genetic diversity can cause not only a decline in certain species, but a decline in the overall productivity and health of an entire ecosystem. Introduction The Peconic Estuary lies between the North and South Forks of Long Island. In recent years, the re-occurrence of Brown Tide algal blooms has put important aquatic organisms in risk for decline. A common organism experiencing such threats is, Eelgrass (represented in our study). The species is the most ecologically significant species of bay-bottom plant in the Peconic Estuary system. It is important that studies such as this are continually performed to monitor the stability and progress of Eelgrass and other organisms. The Forge River is located on the North Shore of Long Island. This river used to be a center for recreation and leisure activities but recently it has been experiencing major changes. The water quality has been significantly degraded due to increased pollution and recent episodes of Hypoxia. Such conditions put native species in risk. For this experiment, we selected a diverse sampling of aquatic plant species living on and near the Forge river and Peconic Estuary. We then sequenced the DNA using the RbCL and tuFa genes to identify the genus of each species. IMAGE 1: Image of barcoded DNA sequence of the species we collected. Used the primer RbCL. IMAGE 2: Image of barcoded DNA sequence of the species we collected. Used the primer tuFa. IMAGE 3: Image of the dna alignment *highlighted species indicate the species we outlined in the discussion. Results Materials & Methods 12 samples of plants growing in the Peconic Estuary and the Forge River area were collected DNA was isolated from each plant sample; it was placed into a tube with lysis solution, grinded up, incubated for 10 minutes, and then centrifuge for 1 minute Then the supernatant was transferred to a new tube and silica resin was added, mix and incubate for 5 minutes, centrifuge for 30 seconds After that remove supernatant, add wash buffer, vortex, centrifuge for 30 seconds, remove supernatant, add wash buffer, vortex, centrifuge for 30 seconds, remove the remaining supernatant Then add dH2O and mix by pipetting in and out, incubate for 5 minutes, centrifuge for 30 seconds, transfer supernatant to new tube, store at -20°C Amplify the DNA by PCR; add primer mix into the tube, then add DNA, add mineral oil, amplify in thermal cycler, store at -20°C Analyze PCR products by gel electrophoresis; pour gel, let it set for 20 minutes, load gel with RCBL and TUFA genes, electrophorese for 30 minutes at 130 volts Sequence PCR product and analyze results; send sample for sequencing, analyze results using bioinformatics DNA subway was used to create a barcode and a phylogenetic tree