Development of microsatellite markers for Pythium helicoides.
A strategy combining dual-suppression PCR and thermal asymmetric interlaced PCR was used to determine sequences flanking microsatellite regions in Pythium helicoides. The primer pairs were designed to amplify loci containing (AC)n, (GA)n, (AGC)n, (CAC)n(CAA)n, (TCA)n and (CTTT)n repeats from the P. helicoides nuclear genome. The PCR products of each primer pair, amplified from three representative isolates collected from different hosts and locations, were cloned and sequenced. Different degrees of polymorphism were detected among these microsatellite markers. The numbers of alleles were 6, 2, 4, 11, 4 and 4 in YL-AC, YL-AGC, YL-CAA, YL-CTTT, YL-GA and YL-TCA, respectively. Allele analysis of 30 P. helicoides isolates showed length polymorphisms in all loci, except for YL-AC, using capillary electrophoresis. Thus, we have developed a simple method for designing PCR primers to amplify microsatellite markers from P. helicoides.
Yin-Ling
The United Graduate School of Agricultural Science, Gifu University, Gifu, Japan. yinling@green.gifu-u.ac.jp
Zhou
Wei
W
Motohashi
Keiichi
K
Suga
Haruhisa
H
Fukui
Hirokazu
H
Kageyama
Koji
K
D017931:DNA Primers; D005828:Genetics, Population; D007564:Japan; D031962:Kalanchoe / Q000382:microbiology; D018895:Microsatellite Repeats / Q000235:genetics*; D010935:Plant Diseases / Q000382:microbiology*; D018517:Plant Roots / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D011775:Pythium / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D027826:Rosa / Q000382:microbiology; D017422:Sequence Analysis, DNA
D017931:DNA Primers