title: Direct DNA extraction method of an obligate parasitic fungus from infected plant tissue.
abstract: Powdery mildew and rust fungi are obligate parasites that cannot live without host organisms. They are difficult to culture in synthetic medium in the laboratory. Genomic DNA extraction is one of the basic molecular techniques used to study the genetic structure of populations. In this study, 2 different DNA extraction methods, Chelex-100 and cetyltrimethylammonium bromide (CTAB), were used to extract DNA from euonymus powdery mildew and Puccinia striiformis f. sp Tritici. Polymerase chain reaction was carried out with a race-specific-marker rDNA-internal transcribed spacer sequence. Both DNA extraction methods were compared and analyzed. The results showed that both Chelex-100 and CTAB were effective for extracting genomic DNA from infected plant tissue. However, less DNA was required for the Chelex-100 method than for the CTAB method, and the Chelex-100 method involved fewer steps, was simpler and safer, and did not require organic solvents compared to the CTAB method. DNA quality was evaluated by polymerase chain reaction, and the results showed that genomic DNA extracted using the Chelex-100 method was better than that using CTAB method, and was sufficient for studying the genetic structure of population.
authors.affiliation: Key Laboratory of Agro-Biodiversity and Pest Management of Education Ministry of China, Yunnan Agricultural University, Kunming, Yunnan, China.
authors.forename: L
authors.initials: L
authors.lastname: Liu
authors.affiliation: Key Laboratory of Agro-Biodiversity and Pest Management of Education Ministry of China, Yunnan Agricultural University, Kunming, Yunnan, China.
authors.forename: C L
authors.initials: CL
authors.lastname: Wang
authors.affiliation: Key Laboratory of Agro-Biodiversity and Pest Management of Education Ministry of China, Yunnan Agricultural University, Kunming, Yunnan, China.
authors.forename: W Y
authors.initials: WY
authors.lastname: Peng
authors.affiliation: Key Laboratory of Agro-Biodiversity and Pest Management of Education Ministry of China, Yunnan Agricultural University, Kunming, Yunnan, China.
authors.forename: J
authors.initials: J
authors.lastname: Yang
authors.affiliation: Key Laboratory of Agro-Biodiversity and Pest Management of Education Ministry of China, Yunnan Agricultural University, Kunming, Yunnan, China.
authors.forename: M Q
authors.initials: MQ
authors.lastname: Lan
authors.affiliation: Key Laboratory of Agro-Biodiversity and Pest Management of Education Ministry of China, Yunnan Agricultural University, Kunming, Yunnan, China.
authors.forename: B
authors.initials: B
authors.lastname: Zhang
authors.affiliation: Agricultural Environment and Resources Research Institute, Yunnan Academy of Agricultural Sciences, Kunming, Yunnan, China.
authors.forename: J B
authors.initials: JB
authors.lastname: Li
authors.affiliation: Key Laboratory of Agro-Biodiversity and Pest Management of Education Ministry of China, Yunnan Agricultural University, Kunming, Yunnan, China.
authors.forename: Y Y
authors.initials: YY
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Agro-Biodiversity and Pest Management of Education Ministry of China, Yunnan Agricultural University, Kunming, Yunnan, China.
authors.forename: C Y
authors.initials: CY
authors.lastname: Li
mesh_terms: D000818:Animals; D004271:DNA, Fungal / Q000302:isolation & purification*; D005658:Fungi / Q000145:classification* / Q000235:genetics*; D010271:Parasites / Q000382:microbiology*; D010641:Phenotype; D010935:Plant Diseases / Q000382:microbiology* / Q000469:parasitology*
chemical_list: D004271:DNA, Fungal