The rapid assignment of ruminal fungi to presumptive genera using ITS1 and ITS2 RNA secondary structures to produce group-specific fingerprints.
Identification of microbial community members in complex environmental samples is time consuming and repetitive. Here, ribosomal sequences and hidden Markov models are used in a novel approach to rapidly assign fungi to their presumptive genera. The ITS1 and ITS2 fragments from a range of axenic, anaerobic gut fungal cultures, including several type strains, were isolated and the RNA secondary structures predicted for these sequences were used to generate a fingerprinting program. The methodology was then tested and the algorithms improved using a collection of environmentally derived sequences, providing a rapid indicator of the fungal diversity and numbers of novel sequence groups within the environmental sample from which they were derived. While the methodology was developed to assist in investigations involving the rumen ecosystem, it has potential generic application in studying diversity and population dynamics in other microbial ecosystems.
Tuckwell
Danny S
DS
F2G Ltd, Lankro Way, Eccles, Manchester M30 0BH, UK.
Nicholson
Matthew J
MJ
School of Biological Sciences, University of Manchester, Oxford Road, Manchester M13 9PT, UK.|Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth, Ceredigion, SY23 3EB, UK.
McSweeney
Christopher S
CS
CSIRO Livestock Industries, Queensland Bioscience Precinct, Carmody Road, St Lucia, Brisbane, Australia.
Theodorou
Michael K
MK
Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth, Ceredigion, SY23 3EB, UK.
Brookman
Jayne L
JL
Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth, Ceredigion, SY23 3EB, UK.
D000818:Animals; D001483:Base Sequence; D016172:DNA Fingerprinting / Q000379:methods*; D004271:DNA, Fungal / Q000032:analysis; D021903:DNA, Ribosomal Spacer / Q000032:analysis*; D005658:Fungi / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D008390:Markov Chains; D008969:Molecular Sequence Data; D016533:Mycological Typing Techniques; D009690:Nucleic Acid Conformation; D012417:Rumen / Q000382:microbiology*; D012418:Ruminants / Q000382:microbiology; D017422:Sequence Analysis, DNA; D013997:Time Factors
D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer