whats new is old resolving the identity of leptothrix ochracea using single cell genomics, pyrosequencing and fish新的是旧解决纤毛菌属的身份ochracea使用单细胞基因组学,焦磷酸测序和鱼.pdfVIP

What’s New Is Old: Resolving the Identity of Leptothrix ochracea Using Single Cell Genomics, Pyrosequencing and FISH 1 1,2 1 1 Emily J. Fleming *, Amy E. Langdon , Manuel Martinez-Garcia , Ramunas Stepanauskas , Nicole J. 1 1 1 Poulton , E. Dashiell P. Masland , David Emerson 1 Bigelow Laboratory for Ocean Sciences, West Boothbay Harbor, Maine, United States of America, 2 Department of Biology, Swarthmore College, Swarthmore, Pennsylvania, United States of America Abstract Leptothrix ochracea is a common inhabitant of freshwater iron seeps and iron-rich wetlands. Its defining characteristic is copious production of extracellular sheaths encrusted with iron oxyhydroxides. Surprisingly, over 90% of these sheaths are empty, hence, what appears to be an abundant population of iron-oxidizing bacteria, consists of relatively few cells. Because L. ochracea has proven difficult to cultivate, its identification is based solely on habitat preference and morphology. We utilized cultivation-independent techniques to resolve this long-standing enigma. By selecting the actively growing edge of a Leptothrix-containing iron mat, a conventional SSU rRNA gene clone library was obtained that had 29 clones (42% of the total library) related to the Leptothrix/Sphaerotilus group (#96% identical to cultured representatives). A pyrotagged library of the V4 hypervariable region constructed from the bulk mat showed that 7.2% of the total sequences also belonged to the Leptothrix/Sphaerotilus group. Sorting of individual L. ochracea sheaths, followed by whole genome amplification (WGA) and PCR identified a SSU rRNA sequence that clustered closely with the putative Lepto