The differentiation of Phytophthora species that are pathogenic on potatoes by an asymmetric PCR combined with single-strand conformation polymorphism analysis.
The goal of this study was to develop a polymerase chain reaction (PCR) capable of differentiating Phytophthora species that are pathogenic on potatoes using a single primer pair. To achieve this objective, primers were derived from conserved regions flanking variable sequences in the internal transcribed spacer 1 (ITS1) of Phytophthora species. One primer pair produced a 140 bp product from P. infestans, P. erythroseptica and P. nicotianae. The PCR products were purified and used in an asymmetric PCR (A-PCR) protocol to generate single-strand DNA (ssDNA). The ssDNA of the Phytophthora potato pathogens reproducibly migrated in non-denaturing polyacrylamide gels in a species-specific manner.
Scott
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DL
US Department of Agriculture, Agriculture Research Service, Beltsville, MD 20705, USA. dscott1906@aol.com
Clark
C W
CW
Fyffe
A E
AE
Walker
M D
MD
Deah
K L
KL
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D004277:DNA, Single-Stranded / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010838:Phytophthora / Q000145:classification / Q000235:genetics* / Q000472:pathogenicity*; D010935:Plant Diseases / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D011198:Solanum tuberosum / Q000382:microbiology; D013045:Species Specificity; D014774:Virulence / Q000235:genetics
D017931:DNA Primers; D004271:DNA, Fungal; D004277:DNA, Single-Stranded