• Parasites, Hosts and Diseases
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• v.33 no.4
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• pp.341-348
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• 1995

Genetic status of Acnnthamoebc sap. were tested on the basis of random amplified polymorphic DNA (RAPD) marker analysis. Four previously established Accnthcmoebn species, 4 Korean isolates of Acnnthamoeba sp., and one American isolate of Acanthcmoebc sp. were analyzed by RAPD-PCR using an arbitrary decamer primers. Amplification products were fractionated by agarose gel electrophoresis and slainrd by ethidium bromide . Eighteen primers produced DNA amplification profiles revealing clear differences among 4 species. Nine of them also produced DNA amplification profiles which included some isolate-specific amplification products. On the basis of amplified fragments by 18 primers, the pairwise similarity indices between A. culbensoni and other species (i.e. A. hntchetti, A. trinngularis, A. polyphaga) were 0.300, 0.308, and 0.313, respectively. Similarity index between A. hctchetti and A. triansulcris was 0.833. The mean similarity index among the 3 Korean isolates (YM-2, -3, -4) was 0.959 and 0.832 among them and 2 other species (A. hatchetti and A. triongulnris). The mean similarity index among YM-5 and other Korean isolates (YM-2, -3, -4) was 0.237. However, the similarity index between YM-5 and A. culbeksoni was 0.857, which suggests that YM-5 is genetically more similar to A. culbertsoni than other Korean isolates. Phonogram reconstructed by UPGMA method revealed that there are two groups: one group consists of A. hctchetti, A. tlonsulcns, and 3 Korean isolates (YM-2, -3, -4) , and the other group consists of A. cuLbensoni. A. polwphosc, HOV, and YM-5.

• Korean journal of applied entomology
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• v.37 no.1
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• pp.31-37
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• 1998

Random amplified polymorphic DNA (RAPD) markers were used to analyze genetic similarity among 8 clones of apierous green peach aphid, two types (M. persicae Sulzer and M. nicotianae lack man) classified by their mo~hologi~cahla raters and host preference (Blackman, 1987), collected from tobacco plants. The genetic variation among these clones was evaluated by polymerase chain reaction amplification with 20 random primers. The higher GC contents of primers, the better in amplification efficiency of PCR reaction in general. The genetic similarities among eight aphid clones were analyzed from UPGMA (unweighted pair group average method) cluster analysis based on simple matching coefficient. The range of genetic similarity coefficients was 0.414 to 0.808. The most close relationship among the clones was similarity coefficient of 0.808 between the PG2 and the PG3 clone. The eight aphid clones analyzed were clustered into three groups by the genetic similarity coefficient. The first group, PG1, PG2, PG3 clone including in M. persicae type by their morphological characters and RED clone in M. nicotianae type was clustered at the genetic similarity coefficient of 0.643. The second group, GR1, GR2, BRN in M. nicotianae type was at the 0.636;and the third group was DBR clone in M. persicae type. The results did not indicate any correlation between m&-phological types (M. persicae and M. nicotianae) and RAPD polymorphism. We could not detect any obvious genetic relationships of the two morphological types of the green peach aphid collected from tobacco plants.

• FLOWER RESEARCH JOURNAL
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• v.19 no.4
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• pp.231-237
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• 2011

In this study, we performed ovule culture after reciprocal crosses of two Alstroemeria accessions and investigated genetic contribution of parents by using RAPD markers. The best method was half-ovule culture on MS medium supplemented with $60g{\cdot}L^{-1}$ sucrose and $2.2g{\cdot}L^{-1}$ gelrite at 14 days after pollination. Embryos began to germinate after 6 weeks of culture. The complete plantlets were formed after 4 months of culture. In eight progenies and two parental cultivars, 59 polymorphic bands were obtained out of 89 total bands by RAPD analysis using 7 primers. Eight $F_1$ progenies from the crosses between two accessions using reciprocal crosses showed 1:1 contribution of maternal and paternal parents. It is confirmed that $F_1$ progenies were obtained from parental accessions by using RAPD markers. We conclude this cross combination showed pre-fertilization barriers with incompatibility between stigma or style, and pollen because progeny number was different in each cross combination. Thereby, it warrants overcoming pre-fertilization barrier together with post-fertilization barrier in order to broaden the heterozygosity within progeny populations in Alstroemeria breeding program.

• Journal of Life Science
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• v.11 no.1
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• pp.65-67
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• 2001

In order to develop a rapid and simple method for testing the purity of radish hybrid seeds using a procedure based on the PCR(Polymerase chain reaction), eighty random primers were screened with the genomic DNA extracted from five day old seedlings of inbred parent lines and their F1 hybrids. Two primers, HRM-02 (5'-GAGACCAGAC-3') and HRM-19(5'-TGAGGCGTGT-3'), generate reproducible unique PCR patterns which can identify each parent lines as well as their hybrids. In actual test of randomly selected hybrid seeds using the two marker primers, the purity tested by one primer was exactly same as that of other primer. It suggests that one marker primer selected in this experiment is enough for the purity test of radish hybrid seeds. We demonstrates the use of RAPD(randomly amplified polymorphic DNAs) markers to identify each of inbred parent lines and hybrids by rapid and simple method.

• The Plant Pathology Journal
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• v.26 no.1
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• pp.83-89
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• 2010

Taxonomic position of 46 Korean isolates of Rhizoctonia solani which were classified into nine intraspecific groups by anastomosis and cultural characteristics was analyzed by randomly amplified polymorphic DNA (RAPD) and sequence analyses of the internal transcribed spacer (ITS) regions of ribosomal DNA. All the isolates within each group showed highly similar band patterns in RAPD. The ITS regions of the isolates within the same groups showed a high level of sequence similarity above 96.0% whereas similarities among different groups were below 94.4%. When compared with several reference strains of R. solani from foreign countries, all the Korean isolates were clustered with the foreign isolates belonging to the same groups in the phylogenetic tree. All six Korean strains of AG-4 were identified as HG-1 out of 3 subgroup of AG-4. We discussed taxonomic position of Korean isolates of R. solani and showed that sequence analysis with ITS regions could be a rapid and useful method for identification of intraspecific group of R. solani.

• Korean Journal of Agricultural Science
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• v.23 no.1
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• pp.99-107
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• 1996

Eighteen nuclear probes were used to examine RFLP(restriction fragment length polymorphism) between six species of Artemisia spp. of Korea. Total DNA from six different species of Artemisia was separately cut with three restrict enzymes. The PstI enzyme was showed to reduce the variation of polymorphisms than the other two enzymes(EcoRl and BamHI). The genetic variation of polymorphism was similar between the Dhewegiki-ssug and Cham-ssug. RAPD analysis was applied to the same six species of Artemisia spp. in order to assess the degree of DNA polymorphism within the Artemisia genus. Six species of Artemisia were evaluated for variation using a set of 11 random 10-mer primers. Nine out of the eleven primers revealed scorable polymorphisms between six species of Artemisia spp. Genetic distances between each of the species were calculated and cluster analysis was used to generate a dendrogram showing phylogenetic relationships between them This result indicates that molecular markers will be more usable in intraspecific study of Artemisia spp. than isoenzyme markers.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.56 no.1
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• pp.35-43
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• 2011

Thirty five isolates of Trichoderma species collected from seven different locations of Bangladesh were studied for morphological characters and molecular variation. Mycelial diameters of the isolates varied from 8.28 cm to 9.00 cm. Based on colony colour, isolates were grouped into five such as dark green, green, light green, yellowish green and whitish green. Maximum isolates were green and light green. On the basis of growth habit and colony consistency, the isolates were categorized into three groups, in which most species had fast growth and were compact in appearance. PCR-based Random Amplified Polymorphic DNA (RAPD) technique employing 3 decamer primers produced 36 scorable bands of which all (100%) were polymorphic. The co-efficient of gene differentiation (Gst) was 1.0000 reflecting the existence of high level of genetic diversity among the isolates. The Unweighted Pair Group Method of Arithmetic Means (UPGMA) dendrogram constructed from Nei's (1972) genetic distance produced 2 main clusters (13 isolates in cluster 1 and 22 isolates in cluster 2). The result indicating their genetic diversity has opened new possibility of using the most efficient and more isolates of Trichoderma in the preparation of biopesticide and decomposition of municipality waste.

• Food Science of Animal Resources
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• v.26 no.3
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• pp.394-401
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• 2006

Forty Clostridium perfringens isolates were obtained from twelve animal products, following the examination of eighty six beef, pork, broiler chicken and salami meat products, and eleven milk powder products. There were 21 isolates from salami stored at $25^{\circ}C$, 3 isolates from pork, 4 isolates from beef, 9 isolates from broiler chicken, and 3 isolates from milk powder. Only the cpa gene encoding a toxin among the 5 toxin genes tested (cpa, cpb, etx, iap, and cpe) was detected in all forty isolates, suggesting contamination with C. perfringens type A. DNA fingerprinting analysis using PCR of the tRNA intergenic spacer (tDNA-PCR) and the 16S-23S internal transcribed spacer (ITS-PCR), and randomly amplified polymorphic DNA (RAPD) analysis were attempted to differentiate the isolates. RAPD analysis was the most discriminating method among the three PCR analyses. Isolates from the same products tended to show similar RAPD patterns. Antimicrobial susceptibility tests showed that some isolates from broiler chickens had the same antibiogram with multiple resistance to streptomycin, colistin, and ciprofloxacin. Antibiograms were similar between isolates from the same livestock products, but differed considerably between the products.

• Journal of Mushroom
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• v.10 no.2
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• pp.93-99
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• 2012

This study was carried to identify a correct species and asses genetic diversity within the same species of Grifola spp. preserved in Division of applied Microbiology. Contaminated isolates showed different growth rates, morphology and color of hyphae. We have reconstructed the phylogenetic tree of a select group of Grifola spp. using nucleotide sequences of the internal transcribed spacer region(ITS) region. The phylogenetic tree was constructed by using the neighbor-joining method. PELF primers of 20-mer were used to assess genetic diversity of preserved isolates. Sequence analysis showed that four strains were identified completely different nomenclature. According to the analysis of ITS sequences, the genus Grifola clustered into one group, most of which correlated with species-groups identified by RAPD method. Eight isolates included strain GM01 showed high similarity with Grifola frondosa. All isolates were collected in the Japan(GM01, GM02, GM03) was identified as Grifola frondosa and isolates of the China(GM05, GM06, GM08) was identified as Bjerkandera fumosa, Grifola frondosa and Dichomitus squalens, respectively. RAPD analysis of genetic polymorphisms of genus Grifola showed a very different band patterns on the isolat. As the result of RAPD and ITS region sequences analysis for preserved isolates, it seems likely that 4 isolates of Grifola spp. may be need to reclassify or eliminate from preserved catalogue.

• Journal of Mushroom
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• v.9 no.1
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• pp.27-33
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• 2011

This study was carried to identify a correct species and asses genetic diversity within the same species of Trametes spp. preserved in Division of applied Microbiology The morphological and cultural characteristics of preserved strains were observed through microscope and investigated on PDA, respectively. Contaminated isolates showed different growth rates, morphology and color of hyphae. We have reconstructed the phylogenetic tree of a select group of Trametes spp. using nucleotide sequences of the internal transcribed spacer region(ITS) region. The phylogenetic tree was constructed by using the neighbor-joining method. PELF primers of 20-mer were used to assess genetic diversity of preserved isolates. Sequence analysis showed that five strains were different species and six strains were identified completely different nomenclature. According to the analysis of ITS sequences, the genus Trametes clustered into four distinct group, most of which correlated with species-groups identified by RAPD method. Seven isolates included TM 01 strain showed high similarity with Trametes versicolr, TM 07 and TM 10 high similarity with Trametes gibbosa, and TM 05 high similarity with Trametes elegans. But isolates collected in the United States was identified as T. junipericola. T. gibbosa and T. versicolor by RAPD analysis of genetic polymorphisms showed a very different band patterns and these strains showed different band patterns on areas. As the result of RAPD and ITS region sequences analysis for preserved isolates, it seems likely that 11 isolates of Trametes spp. may be need to reclassify or eliminate from preserved catalogue.