Journal of Microbiology

Selection of RAPD Markers for Phytophthora infestans and PCR Detection of Phytophthora infestans from Potatoes: Kyoung Su Kim , Youn Su Lee; J. Microbiol. 2001;39(2):126-132.

Abstract: For rapid and secure differentiation of P. infestans from other Phytophthora species, two fragments obtained from randomly amplified polymorphic DNA (RAPD) profiles were selected as markers. Also, primers for in polymerase chain reaction (PCR) to detect P. infestans specifically were developed by analyzing the sequences of ITSII regions in rDNA of Phytophthora species. The primers, PISP-1 and ITS3 amplified a single. Fragment 450 bp of about in P. infestans , but not in other fungal or bacterial isolates. Annealing temperatures and template DNA quantities were varied for the optimization of PCR conditions. From the result of the PCR detection study, species-specific primers were selected under annealing temperatures ranging from 55 C to 61 C, and template DNA levels ranging from 10 pg to 100 ng.

Genetic DNA Marker for A2 mating type in Phytophthora infestans: Kwon Jong Kim , Youn Su Lee; J. Microbiol. 2002;40(4):254-259.

Abstract: The Phytophthora infestans requires two mating types for sexual reproduction. Amplified fragment length polymorphism (AFLP) was used to specifically detect different mating types of P. infestans . The AFLP primers E+AA (5'-GACTGCGTACCAATTCAA-3') and M+CAA (5'-GATGAGTCCTGAGTAACAA-3') detected a fragment that is specific in the A2 mating type of P. infestans . This fragment was cloned and sequenced. Based on the sequence data, PHYB-1 and PHYB-2 primer were designed to detect the A2 mating type of P. infestans. A single 347 bp segment was observed in the A2 mating type of P. infestans, but not in the A1 mating type of P. infestans or other Phytophthora spp. Identification of mating type was performed with phenotype (sexual reproduction) and genotype (CAPs marker) methods. Two factors, the annealing temperature and template DNA quantity, were investigated to determine the optimal conditions. Using mating type-specific primers, a unique band was obtained within annealing temperatures of 57 ℃-62℃ and DNA levels of 10pg-100 ng (data not shown).

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