• Korean Journal of Veterinary Research
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• v.42 no.3
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• pp.363-370
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• 2002

For the development of diagnostic polymerase chain reaction (PCR) to fungal infection by dermatophytes Trichophyton and Microsporum, detection of the fungal DNA by PCR and analysis of the DNA pattern were undertaken in the present study. A total of 15 strains were tested and those consisted of 3 reference strains and 12 isolates such as: reference strains of T mentagrophytes (downy type, ATCC 9533), T rubrum (IFO 6204) and M gypseum (ATCC 9083), and each isolate of T mentogrophytes (powdery type), T mentagrophytes (granular type), T mentogrophytes (purple-red type), T rubrum, T raubitschekii, T tonsurans, T equinum, T ajelloi, T verrucosum, M cookei, M nanum and M gypseum. The DNA were purely isolated from all strains of Trichophyton spp. and Microsporum spp. by a simple method partly consisted of disruption of fungal cells by lyophilization and grinding and extraction of fungal DNA without phenol treatment which is a routine procedure in DNA isolation. For the detection of fungal DNAs, optimal condition of PCR was determined as preheating once at $94^{\circ}C$ for 5 min, 35 cycles of denaturation at $94^{\circ}C$ for 1 min, annealing at $38^{\circ}C$ for 1 min and polymerization at $72^{\circ}C$ for 2 min, and 1 cycle of final extension at $72^{\circ}C$ for 5 min. In PCR using arbitrary primers AP-1 (5' ACCCGACCTG3') and AP-2 (5' ACGGGCCAGT3'), DNAs in various numbers and sizes were detected from different species of Trichophyton and Microsporum, while DNAs in similar size were also detected in all strains of Trichophyton spp. and Microsporum spp. There were unique DNAs observed from certain dermatophytes by AP-1 such as 1,900 bases in T rubrum, 950 and 1,100 bases in T raubitscheldi, 2,100 bases in T equinum, 400 bases in T verrucosum and 1,150 bases in M gypseum. The unique DNAs were also observed by AP-2 such as 1,200 bases in T ajelloi, 250 bases in T verrucosum, 1,150 bases in M cookei and 2,000 bases in M nanum. The results indicated that PCR can detect a specific DNA from certain Trychophyton and Microsporum spp, which can be the information for further development of diagoomc PCR to dennatophytes.

• The Journal of the Korean Society for Microbiology
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• v.35 no.1
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• pp.19-29
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• 2000

Although most persons infected with Helicobacter pylori harbor a single strain of the organism, multiple strain colonization in the same patient is also occasionally reported in developed countries. The aims of this study were to determine the prevalence of multiple strain colonization in Korean patients and to detect the cagA, iceA1, and babA status of H. pylori isolated from the antrum and body of the stomach. H. pylori was obtained from 35 patients from the antrum and body of the stomach. The genomic diversity of H. pylori was determined by random amplified polymorphic DNA analysis. The status of cagA, iceA1, and babA genes of H. pylori was assessed by polymerase chain reaction with appropriate primers. Clearly different diversity patterns were identified among the isolates from 35 individual patients. Eighteen (51.4%) patients had a single strain of H. pylori. Eight (22.9%) and nine (25.7%) patients had subtypically (one or two bands difference) and typically (clearly different pattern) different strains of H. pylori in the antrum and body, respectively. Among the 70 isolates of H. pylori from 35 patients, the positive rates of 349-bp and 208-bp cagA gene fragments and the iceA1 gene were 68/70 (97.1%), 68/70 (97.1%), and 58/70 (82.9%), respectively. However, the babA gene was found in 22/66 cases (31.4%). In five out of 18 patients with a single strain, the genetic status of cagA, iceA1, and babA varied between the isolates from the antrum and the body. In 8/17 patients with sub typically or typically different strains, the gene status differed between antrum and body isolates. The prevalence of co-colonization with typically or subtypically different strains is high in Korea, and sub-clones with different pathogenic gene status exist within strains of identical RAPD patterns.

• Korean Journal of Plant Resources
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• v.17 no.2
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• pp.107-115
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• 2004

To develop universal primers for phylogenetic analysis and species-specific marker for breeding program of kiwifruit, eighteens primers were designed from kiwifruit genome-specific repeat sequences. Seven species including twenty two varieties collected from native eastern Asia were examined using 18 to 22 mer kiwifruit target(KT) primers. among eighteen primers, we selected seven primers for phylogenetic relationship. The genus Actinidia was divided into two large groups; group I,A. arguta, A. melanandra, A. kolomikta, and A. marcrosperma, characterized by the non-hair in fruits and loaves or a few pubescences only in young stage, which belongs to the section Leiocarpae, and group II, A. chinensis, A. deliciosa, and A. eriantha, characterized by a lot of hairs only in young fruit stage and with a lot of hairs or fuzzes in leaves and branches, which belongs to the section Stellatae. Group II especially belongs to the series Perfectae of the section Stellatae and was divided into two subgroups; subgroup I containing A. chinensis and A. deliciosa, and subgroup II containing A. eriantha. In contrast, the two species, A. chinensis and A. deliciosa, which are known to have common parents, were divided into two independent subgroups with 80％ of a similarity value. On the other hand, we selected KT6F for variety specific bands, KT12E primers for 'Hayward' and 'Tomuri'. KT7F or KT12F primers were useful for analysis of inheritance pattern in kiwifruit cross-breeding. We suggest that these primers will be a powerful tool for elucidating phylogenetic relationship and selection of novelty kiwifruit in a breeding program.

• Journal of Mushroom
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• v.19 no.2
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• pp.88-95
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• 2021

This study was carried out to breed new variety of Pleurotus nebrodensis. We have collected and tested characteristics of genetic resources from domestic and abroad since 2015. The varieties of P. nebrodensis from China are grown by farmers, but those have been unstable fruiting and are weak against bacterial diseases. To solve this problem, we bred the unique domestic variety 'Uram' of P. nebrodensis and the results of the characteristic test for the new 'Uram' are as follows. The proper temperature for the mycelial growth was 26~29℃ and fruit body growth temperature was 15~18℃. It was similar to the control variety KME65035 of P. nebrodensis in the pileus form of a flat and white color. The number of days required for initial fruting was 5 days for bottle cultivation and 6 days for bag cultivation which was 2-4 days shorter than that of the control variety. The pileus diameter was 32.6-37.0 mm which was smaller but the fruit body length was 130.4 mm, which was longer than those of the control variety. The effective number of fruit bodies was 1.8 in bottle cultivation and 2.9 in bag cultivation, which was more than those of the control variety. The yield rate was 93.3-100%, which was more stable than those of the control variety. In bottle cultivation and bag cultivation, the yield was 173.1 g/bottle (1100 cc) and 283.4 g/bag (1.2 kg), respectively, which was 25-44% higher than those of the control variety 138.0 g/bottle (1100 cc) and 197.4 g bag (1.2 kg). When incubating the parent and control varieties of 'Uram', the replacement line was clear and as a result of mycelial DNA RAPD-PCR reaction, the band pattern was different from that of the parent and control varieties, confirming the hybrid species.