Exploring microbial diversity and taxonomy using SSU rRNA hypervariable tag sequencing
Huse, Susan M.
Huber, Julie A.
Mark Welch, David B.
Relman, David A.
Sogin, Mitchell L.
MetadataShow full item record
Massively parallel pyrosequencing of hypervariable regions from small subunit ribosomal RNA (SSU rRNA) genes can sample a microbial community two or three orders of magnitude more deeply per dollar and per hour than capillary sequencing of full-length SSU rRNA. As with full-length rRNA surveys, each sequence read is a tag surrogate for a single microbe. However, rather than assigning taxonomy by creating gene trees de novo that include all experimental sequences and certain reference taxa, we compare the hypervariable region tags to an extensive database of rRNA sequences and assign taxonomy based on the best match in a Global Alignment for Sequence Taxonomy (GAST) process. The resulting taxonomic census provides information on both composition and diversity of the microbial community. To determine the effectiveness of using only hypervariable region tags for assessing microbial community membership, we compared the taxonomy assigned to the V3 and V6 hypervariable regions with the taxonomy assigned to full-length SSU rRNA sequences isolated from both the human gut and a deep-sea hydrothermal vent. The hypervariable region tags and full-length rRNA sequences provided equivalent taxonomy and measures of relative abundance of microbial communities, even for tags up to 15% divergent from their nearest reference match. The greater sampling depth per dollar afforded by massively parallel pyrosequencing reveals many more members of the “rare biosphere” than does capillary sequencing of the full-length gene. In addition, tag sequencing eliminates cloning bias and the sequences are short enough to be completely sequenced in a single read, maximizing the number of organisms sampled in a run while minimizing chimera formation. This technique allows the cost-effective exploration of changes in microbial community structure, including the rare biosphere, over space and time and can be applied immediately to initiatives, such as the Human Microbiome Project.
© 2008 Huse et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS Genetics 4 (2008): e1000255, doi:10.1371/journal.pgen.1000255.
The following license files are associated with this item:
Showing items related by title, author, creator and subject.
Feary, David A.; Burt, John A.; Bauman, Andrew G.; Al Hazeem, Shaker; Abdel-Moati, Mohamed A.; Al-Khalifa, Khalifa A.; Anderson, Donald M.; Amos, Carl; Baker, Andrew; Bartholomew, Aaron; Bentok, Rita; Cavalcante, Georgenes H.; Chen, Chaolun Allen; Coles, Steve L.; Dab, Koosha; Fowler, Ashley M.; George, David; Grandcourt, Edwin; Hill, Ross; John, David M.; Jones, David A.; Keshavmurthy, Shashank; Mahmoud, Huda; Moradi Och Tapeh, Mahdi; Mostafavi, Pargol Ghavam; Naser, Humood; Pichonz, Michel; Purkis, Sam; Riegl, Bernhard; Samimi-Namin, Kaveh; Sheppard, Charles; Samiei, Jahangir Vajed; Voolstra, Christian R.; Wiedenmann, Joerg (2013-02)Expert opinion was assessed to identify current knowledge gaps in determining future changes in Arabian/Persian Gulf (thereafter ‘Gulf’) coral reefs. Thirty-one participants submitted 71 research questions that were ...
David, John; Garrity, George M.; Greuter, Werner; Hawksworth, David L.; Jahn, Regine; Kirk, Paul M.; McNeill, John; Michel, Ellinor; Knapp, Sandra; Patterson, David J.; Tindall, Brian J.; Todd, Jonathan A.; Tol, Jan van; Turland, Nicholas J. (Pensoft, 2012-05-08)A set of terms recommended for use in facilitating communication in biological nomenclature is presented as a table showing broadly equivalent terms used in the traditional Codes of nomenclature. These terms are intended ...
Carter, William E.; Aubrey, David G.; Baker, Trevor; Boucher, Claude; LeProvost, Christian; Pugh, David T.; Peltier, W. R.; Zumberge, Mark A.; Rapp, Richard H.; Schultz, Robert E.; Emery, Kenneth O.; Enfield, David B. (Woods Hole Oceanographic Institution, 1989-08)Under the auspices of the International Association for Physical Sciences of the Ocean (IAPSO) a committee was established to identify the oceanographic and geophysical requirements for fixing Tide Gauge Bench Maries ...