Shifts in Microbial Communities Based Upon Transect Habitat By Logan Greydanus A PROJECT Submitted to Oregon State University University Honors College in partial fulfillment of the requirements for the degree of Honors Baccalaureate of Science in Microbiology (Honors Scholar) Presented May 30, 2014 Commencement June, 2014 ! "! AN ABSTRACT OF THE THESIS OF Logan Greydanus for the degree of Honors Scholar of Science in Microbiology presented on May 30th, 2014. Title: Shifts in Microbial Communities Based Upon Transect Habitat Abstract Approved: Rebecca Vega-Thurber Coral reef health has been in severe decline around the globe in the past several decades, in many cases due to direct human impact. Human action, such as overfishing, habitat destruction and nutrient loading, has caused coral coverage to drop to record lows, threatening the future of these critically important biodiversity hotspots. This study investigates the changes in microbial community along reef transects on the South Pacific island of Moorea. Samples were collected at three distinct habitats along reef transects: Fringing Reef, Back Reef, and Cresting Reef. Reef habitats were compared between three regions around the island: North, East and West. Results indicate that there is a significant difference in Bacterial and Archaeal (16S) species between the inner fringing reef habitat and the outer Cresting and Back reef habitats. This difference can consistently be seen across all regions sampled around Moorea. Comparison of the 16S communities based solely on region resulted in no significant difference, suggesting that the primary driver of microbial diversity is location on transect and not region. Based on these results, it appears the Bacterial and Archaeal communities are defined by the reef habitat they belong to and that these communities are not unique to region. Keywords: Bacteria, marine microbiome, coral reefs, DNA sequencing, 16S Corresponding E-mail address: logangreydanus@gmail.com ! #! ! Copyright Logan Greydanus May 30th, 2014 All Rights Reserved ! $! Shifts in Microbial Communities Based Upon Transect Habitat By Logan Greydanus A PROJECT Submitted to Oregon State University University Honors College in partial fulfillment of the requirements for the degree of Honors Baccalaureate of Science in Microbiology (Honors Scholar) Presented May 30, 2014 Commencement June, 2014 ! %! Honors Scholar of Science in Microbiology project of Logan Greydanus presented on May 30th, 2014. APPROVED: Mentor, representing Microbiology Committee Member, representing Microbiology Committee Member, representing Microbiology Dean, University Honors College I understand that my project will become part of the permanent collection of Oregon State University, University Honors College. My signature below authorizes release of my project to any reader upon request. Logan Greydanus, Author ! &! Table of Contents Introduction: …………………………………………………………………………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ist of Figures Figure 1: Moorea…………………………………………………………………... 10 Table 1: PCR Reagent Concentrations and Volumes ……………………………... 12 Figure 2: Taxa Summary Filtered by Region at the Family Level ………………... 16 Figure 3: Taxa Summary Filtered by Reef Habitat at the Family Level.………….. 16 Figure 4: MDS Plots………………………………………………………………. 18 Figure 5: Genera Level Top 5……………………………………………………... 21 ! B! ! 7! "#$%&'!$(!)$*+,-$./!0,112($&$3'!4.'35!67,(!8+.('3*&!9.-$&.&! :(&+,52*&$,(;! =2,).!,++>3!,+@,+3+/*!*1+!D23*!0-E+,3+!D),-/+!+<23F3*+D3!2/!*1+!@.)/+*6! ?.*129G1!*1+F!D)H+!9@!)!3D)..!@+,<+/*)G+!2>!*1+!*2*).!D),-/+!1)I-*)*J!*1+!0-E+,3-*F! 2>!.->+!)/0!/9DI+,!2>!+<2.2G-<).!,2.+3!>-..+0!-3!*,+D+/02936!K9D)/3!0+,-E+!G,+)*! 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