• Journal of Mushroom
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• v.10 no.1
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• pp.37-43
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• 2012

This study was carried to identify a correct species and asses genetic diversity within the same species of Polyporus 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 Polyporus 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 three strains were different species and four strains were identified completely different nomenclature. According to the analysis of ITS sequences, the genus Polyporus clustered into five distinct group, most of which correlated with species-groups identified by RAPD method. Four isolates included strain PM02 showed high similarity with P. arcularius, four isolates included strain PM03 high similarity with P. alveolaris, three isolates included strain PM01 high similarity with P. tuberaster, and PM 06 and PM04 high similarity with P. brumalis and P. squamossus. Isolates were collected in the United States(PM10, PM11) was identified as P. alveolarius and P. arcularius. RAPD analysis of genetic polymorphisms of genus Polyporus showed a very different band patterns. As the result of RAPD and ITS region sequences analysis for preserved isolates, it seems likely that 6 isolates of Polyporus spp. may be need to reclassify or eliminate from preserved catalogue.

• Korean Journal of Breeding Science
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• v.41 no.4
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• pp.437-443
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• 2009

In this study, we used the random amplified polymorphic DNA (RAPD) technique to evaluate the genetic relationships among 29 grapevine cultivars (Vitis spp.). Sixty selective primers detected a total of 558 polymorphic bands. By UPGMA (unweighted pair-group method arithmetic average) cluster analysis with 558 polymorphic bands, the 29 grapevine cultivars were divided into six major groups at 58.8% genetic similarity. The "Super Hamburg" was clustered in group I. Group II consisted of "Wonkyo RA-23", "Muscat Hamburg", "Tano Red", and "Tankeumchu". Group III consisted of "Alden", "Wonkyo RA -21", "Wonkyo RA-30", and "Dutchess". Group IV included 14 grapevine cultivars ("Heukgoosul", "Heukbosuk", "Suok", "Wonkyo RA-29", "Wonkyo RA-22", "Kyoho", "Pione", "Beniizu", "Golden Muscat", "Jinok", "Doonuri", "Campbell Early", "Delaware", and "Schuyler"). Group V consisted of "Hongdan", "Tamnara", "Hongisul", and "Himrod seedless". Group VI included 2 cultivars ("Cheongsoo", and "S. 9110").

• Korean Journal of Medicinal Crop Science
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• v.10 no.3
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• pp.194-199
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• 2002

Extracted genomic DNA from 16 accessions of Codonopsis lanceolata collected from South Korea and the Baekdoo Mt. areas of China were analyzed for their genetic relationships by RAPD. Twenty 10-mer-oligonucleotide primers having reproductive polymorphism were selected for the RAPD analysis. The size of amplified DNA was almost between 125 bp and 2.0 kbp. Sixteen collected Codonopsis lanceolata were analyzed with 20 primers which generated 73(49.3%) polymorphic bands among 148 PCR products. The mean number of polymorphic bands were 7.4 and varied $1{\sim}9$ per primer. It was, thus, demonstrated that RAPD was useful for detecting polymorphism in Codonopsis lanceolata. The range of 1-F value(genetic similarity) was from 0.682 to 0.959. These results indicate variable genetic similarities. By UPGMA (Unweighted Pair Group Method using an Arithmetic average) cluster analysis based on 1-F value, genetic distance among the 16 collected Codonopsis lanceolata was $0.133{\sim}0.400$. It was certainly classified into two groups between collected accessions from Korea and China, and the genetic distance was about 0.281. Both accessions collected from Korea and China showed miner differences, while the genetic relationships of Tonghua Xian and Liuhe Xian from China was farthest with other accessions collected.

• Journal of Korean Society of Forest Science
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• v.87 no.3
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• pp.422-428
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• 1998

Polymerase chain reaction(PCR)-based inter-simple sequence repeats(I-SSR) markers were analyzed in 48 megagametophytes of a single tree of Abies koreana $W_{ILS}$. Nineteen of the 35 primers, screened with 6 megagametophyte DNA and produced the clearest amplification products in the preliminary experiment, were used for PCR with 48 megagametophyte DNAs sampled from a single tree. On the basis of the chi-square test, a total of 51 amplicons, amplified by the 19 primers, were revealed to be segregated according to the Mendelian ratio(i.e., 1 : 1 segregation ratio) in the 48 megagametophytes at 5% significance level. Based on the linkage analysis, the observed 51 Mendelian loci turned out to be unlinked each other, which suggested that they are evenly distributed in the genome. However, majority of RAPD markers are known to belong to the independent linkage blocks, which frequently results in the amplification of RAPD markers from the restricted regions of the genome. Owing to the nature of even distribution of the 51 loci observed in this study, the I-SSR markers could give better resolution of estimating genetic diversity from the whole genome than RAPD markers. And I-SSR markers are also more suitable than RAPD markers for reconstructing phylogenetic relationship by a cladistic method which requires to fulfil the assumption of independent evolution of the different characters.

• Korean Journal of Medicinal Crop Science
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• v.10 no.1
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• pp.17-23
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• 2002

The seeds of Coix lachryma-jobi Linne var. mayuen Stapf. are used as dietary food for obesity and diabetes under the names of Yulmu in Korea and Yiyiren(薏苡仁) in China. It is one of the drugs promoting diuresis to eliminate the wetness-evil from the lower warmer in the traditional Korean medicine. According to ancient textbook of the traditional Korean medicine, it should be applied to patients with phlegm and heat, etc. The establishment of the method for the discrimination of Coisis Semen is very important for the quality control of drugs. Random amplified polymorphic DNA(RAPD) analysis and anatomical characteristics were used for the discrimination of Coix lachryma-jobi $Linn\acute{e}$ var. mayuen $S_{TAPF}$. and C. lachryma-jobi $Linn\acute{e}$. In the RAPD analysis with 20 primers, 8 primers gave informative and reproducible bands with the genomic DNA. From the cluster analysis, the genus Coix were divided into two groups at similarity coefficient of 0.863.

• Korean Journal of Medicinal Crop Science
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• v.15 no.1
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• pp.1-5
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• 2007

To develop a convenient method for discriminating between violet flowered lines and white flowered lines in Chinese bellflower, RAPD analysis was carried out and SCAR markers were generated. Eighteen specific RAPD bands were obtained from 6 OPERON primer sets. Two out of eighteen RAPD bands were cloned into pGEM-T-Easy vectors and then subjected to the nucleotide sequence analysis. PgR1 and PgR2 DNA fragment, each specific for violet and white flowered lines, consist of 887 bp and 863 bp sequences, respectively. Two SCAR markers were developed from RAPD clones: SPgR1 (355 bp) from PgR1 and SPgR2 (493 bp) from PgR2. One (SPgR2) of these two markers was useful to differentiate between violet flowered lines and white flowered lines in Chinese bellflower.

• Journal of Life Science
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• v.18 no.4
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• pp.435-440
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• 2008

Random amplified polymorphic DNA (RAPD) markers were used to identify the genetic diversities among and within varieties and landraces of Rehmannia glutinosa. Polymorphic and reproducible bands were produced by 10 primers out of total 20 primers used in the experiment. In RAPD analysis of the 11 genotypes, 64 fragments out of 73 amplified genomic DNA fragments were polymorphic which represented an average 6.4 polymorphic fragments per primer. Number of amplified fragments with random primers ranged from 2 (OPA-1) to 13 (OPA-11) and varied in size from 200 bp to 1,400 bp. Especially, OPA-10, OPA-11 and OPA-19 primers showed specific bands for varieties of Korea Jiwhang and Jiwhang il ho, which could be useful for discriminating from other varieties and landraces of R. glutinosa. Percentage polymorphism ranged from a minimum of 50% (OPA-1) to a maximum of 100% (OPA-11), with an average of 87.7%. Similarity coefficients were higher in the genotypes of Korea Jiwhang and Jiwhang il ho than in other populations. In cluster analysis, genotypes of Korea Jiwhang, Jiwhang il ho, and Japanese accession were separated from those of other varieties and landraces. Average of genetic diversity within the population $(H_S)$ was 0.110, while average of total genetic diversity $(H_T)$ was 0.229. Across all RAPD makers the $G_{ST}$ value was 0.517, indicating that about 52% of the total genetic variation could be explained by RAPDs differences while the remaining 48% might be attributable to differences among samples. Consequently, RAPD analysis was useful method to discriminate different populations such as domestic varieties and other landraces. The results of the present study will be used to understand the population and evolutionary genetics of R. gllutinosa.

• The Korean Journal of Mycology
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• v.25 no.2 s.81
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• pp.121-129
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• 1997

Tricholoma matsutake (TM=T. caligatum var. nauseoum) is, for an agricultural income earned by Forestry by-products, a very important mushroom in Korea. The mycelia isolated from the basidiocarps were compared with basidiocarps of TM by the random amplified polymorphisms of RAPD-DNA bands. The mycelia were confirmed to be originated from the basidiocarps of TM by cluster analyses of the DNA-bands made from RAPDs and Southern blotting with the band (0.75 kb) identified. The mycelia defined were observed to grow very slowly at the rate of 10 cm per month at $25^{\circ}C$ and also to be semi-transparent and submerged in on PDA. The method developed in this work was considered to be very useful for confirming the mycelia originated from the ectomycorrhizal mushrooms and also to be applied for the fungal mycelia isolated from the commercial useful mushrooms.

• The Plant Pathology Journal
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• v.22 no.1
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• pp.36-45
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• 2006

Twenty isolates of Didymella bryoniae were isolated from infected cucurbit plants in various growing areas of southern Korea in 2001 and 2002. Random Amplified Polymorphic DNA (RAPD) group [RG] I of D. bryoniae was more virulent than RG IV to watermelon. Virulence of the RG I isolate was strong to moderate to cucumber, whereas that of the RG IV varied from strong, moderate to weak. Two hundred seventy-three amplified fragments were produced with 40 primers, and were analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYSPC. At the distance level of 0.7, two major genomic DNA RAPD groups were differentiated among 20 isolates. The RG I included 7 isolates from watermelon and one isolate from melon, whereas the RG IV included 12 isolates from squash, cucumber, watermelon and melon. Amplification of internal transcribed spacer (ITS) region and small subunit rRNA region from the 20 isolates yielded respectively a single fragment. Restriction pattern with 12 restriction enzymes was identical for all isolates tested, suggesting that variation in the ITS and small subunit within the D. bryoniae were low. Amplification of the genomic DNAs of the tested isolates with the sequence characterized amplified regions (SCAR) primer RG IF-RG IR specific for RG I group resulted in a single band of 650bp fragment for 8 isolates out of the 20 isolates. Therefore, these 8 isolates could be assigned into RG I. The same experiments done with RG IIF-RG IIR resulted in no amplified PCR product for the 20 isolates tested. An about 1.4 kb-fragment amplified from the RG IV isolates was specifically hybridized with PCR fragments amplified from genomic DNAs of the RG IV isolates only, suggesting that this PCR product could be used for discriminating the RG IV isolates from the RG I isolates as well other fungal species.

• Journal of the Korean Wood Science and Technology
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• v.34 no.6
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• pp.96-103
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• 2006

A simple method for identifying of ectomycorrhizal fungi was presented, using the polymerase chain reaction (PCR) to amplify the ITS (Internal transcribed spacer) regions of the nuclear ribosormal repeat. The sequences analyzed 6 species, Pisolithus tinctorius, Chroogomphus rutilus, Leucogyrophana pinastri, Suillus granulatus, Lactarius laeticolorus, and Suillus bovinus at hongreung forest, and analysed 10 species, Craterellus lutescens, Thelephoroid mycorrhizal, Lactarius quieticolor, Tricholoma matsutake, Lactarius chrysorrheus, Sarcodon aspratus, Russula versicolor, Suillus luteus, Tricholoma terreum, and Amanita vaginata at hongcheon forest. Finally, the amplification by PCR with ITS1-ITS4 primers offers good results over classical identification for ectomycorrhizal fungi species.