A strategy for recovering high quality genomic DNA from a large number of Phytophthora isolates

被引:39
作者
Lamour, Kurt [1 ]
Finley, Ledare [1 ]
机构
[1] Univ Tennessee, Inst Agr, Dept Entomol & Plant Pathol, Knoxville, TN 37901 USA
关键词
AFLP; oomycetes; reverse genetics; TILLING;
D O I
10.3852/mycologia.98.3.514
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
We present a strategy to recover high molecular weight genomic DNA from large numbers of isolates of Phytophthora. Included are steps for generating mycelial mass in 24-well reuseable deep well plates, efficient lyophilization and disruption of the mycelium and genomic DNA extraction with 96-well glass fiber filter plates. The resulting DNA is consistently high molecular weight and is suitable for applications that require high quality DNA such as AFLP analysis and TILLING. A single operator easily can manage mycelium preparation and/or DNA extraction from 384 isolates in a single day and this approach might be useful for other fungi or fungi-like organisms that can be grown in liquid media.
引用
收藏
页码:514 / 517
页数:4
相关论文
共 10 条
[1]   Use of a single primer to fluorescently label selective amplified fragment length polymorphism reactions [J].
Habera, L ;
Smith, N ;
Donahoo, R ;
Lamour, K .
BIOTECHNIQUES, 2004, 37 (06) :902-+
[2]   Single-nucleotide mutations for plant functional genomics [J].
Henikoff, S ;
Comai, L .
ANNUAL REVIEW OF PLANT BIOLOGY, 2003, 54 :375-401
[3]   Etiology of Phytophthora drechsleri and P-nicotianae (=P. parasitica) diseases affecting floriculture crops [J].
Lamour, KH ;
Daughtrey, ML ;
Benson, DM ;
Hwang, J ;
Hausbeck, MK .
PLANT DISEASE, 2003, 87 (07) :854-858
[4]   Effect of crop rotation on the survival of Phytophthora capsici in Michigan [J].
Lamour, KH ;
Hausbeck, MK .
PLANT DISEASE, 2003, 87 (07) :841-845
[5]   Mefenoxam insensitivity and the sexual stage of Phytophthora capsici in Michigan cucurbit fields [J].
Lamour, KH ;
Hausbeck, MK .
PHYTOPATHOLOGY, 2000, 90 (04) :396-400
[6]   The spatiotemporal genetic structure of Phytophthora capsici in Michigan and implications for disease management [J].
Lamour, KH ;
Hausbeck, MK .
PHYTOPATHOLOGY, 2002, 92 (06) :681-684
[7]   Targeting induced local lesions in genomes (TILLING) for plant functional genomics [J].
McCallum, CM ;
Comai, L ;
Greene, EA ;
Henikoff, S .
PLANT PHYSIOLOGY, 2000, 123 (02) :439-442
[8]   A TILLING reverse genetics tool and a web-accessible collection of mutants of the legume Lotus japonicus [J].
Perry, JA ;
Wang, TL ;
Welham, TJ ;
Gardner, S ;
Pike, JM ;
Yoshida, S ;
Parniske, M .
PLANT PHYSIOLOGY, 2003, 131 (03) :866-871
[9]   Large-scale discovery of induced point mutations with high-throughput TILLING [J].
Till, BJ ;
Reynolds, SH ;
Greene, EA ;
Codomo, CA ;
Enns, LC ;
Johnson, JE ;
Burtner, C ;
Odden, AR ;
Young, K ;
Taylor, NE ;
Henikoff, JG ;
Comai, L ;
Henikoff, S .
GENOME RESEARCH, 2003, 13 (03) :524-530
[10]   Target-selected mutant screen by TILLING in Drosophila [J].
Winkler, S ;
Schwabedissen, A ;
Backasch, D ;
Bökel, C ;
Seidel, C ;
Bönisch, S ;
Fürthauer, M ;
Kuhrs, A ;
Cobreros, L ;
Brand, M ;
González-Gaitán, M .
GENOME RESEARCH, 2005, 15 (05) :718-723