Journal
JOURNAL OF MICROBIOLOGICAL METHODS
Volume 77, Issue 2, Pages 229-234Publisher
ELSEVIER
DOI: 10.1016/j.mimet.2009.02.007
Keywords
beta-tubulin gene; Multiple displacement amplification; rDNA-ITS; Single-spore; Whole genome amplification
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Funding
- National Basic Research Program [2006CB100204]
- National Natural Science Foundation of China [30670013]
- Education Ministry of China [1307049]
- Program for Changjiang Scholars and Innovative Research Team in University [200558, IRT0748]
- Natural Sciences and Engineering Research Council of Canada
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Rust fungi are obligate parasites and cannot be routinely cultured to obtain sufficient biomass for DNA extractions. Multiple displacement amplification (MDA) was demonstrated in this study for whole genome amplification from single spores of the rust fungus, Puccinia striiformis. The genomic DNA coverage and fidelity of this method was evaluated by PCR amplification and sequencing of two genetic markers: portions of the multi-copy nuclear ribosomal DNA internal transcribed spacer region (ITS) and the single copy beta-tubulin gene from two geographical diverse isolates. our results show that MDA is a valuable tool for whole genome amplification from single spores, and we propose that MDA-amplified DNA can be used for molecular genetic analysis of the wheat yellow rust fungus. (C) 2009 Elsevier B.V. All rights reserved.
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