title: Whole genome amplification of the rust Puccinia striiformis f. sp. tritici from single spores.
abstract: 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.
authors.affiliation: College of Plant Protection, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: Yanchun
authors.initials: Y
authors.lastname: Wang
authors.forename: Mingqi
authors.initials: M
authors.lastname: Zhu
authors.forename: Rong
authors.initials: R
authors.lastname: Zhang
authors.forename: Hanli
authors.initials: H
authors.lastname: Yang
authors.forename: Yang
authors.initials: Y
authors.lastname: Wang
authors.forename: Guangyu
authors.initials: G
authors.lastname: Sun
authors.forename: Shelin
authors.initials: S
authors.lastname: Jin
authors.forename: Tom
authors.initials: T
authors.lastname: Hsiang
mesh_terms: D001483:Base Sequence; D001487:Basidiomycota / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D005800:Genes, Fungal; D016681:Genome, Fungal; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013172:Spores, Fungal / Q000235:genetics*; D014908:Triticum / Q000382:microbiology; D014404:Tubulin / Q000235:genetics
chemical_list: D017931:DNA Primers; D021903:DNA, Ribosomal Spacer; D014404:Tubulin