• The Korea Journal of Herbology
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• v.24 no.4
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• pp.115-120
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• 2009

Objectives : Due to the morphological similarity and frequent occurrence of intermediate forms as well as morphological variations of aerial part, the correct identification between Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma is very difficult. To develop a reliable method for correct identification and improving the quality standards of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma, we analyzed RAPD and developed SCAR marker. Methods : To amplify target DNA at the genomic level, 32 Operon 10-mer random primers were applied with four Rheum species, R. officinale, R. palmatum, R. tanguticum and R. undulatum. The nucleotide sequences were determined and species-specific primers were prepared depending on the species-specific RAPD amplicons after subcloned into the pGEM-Teasy vector. To develop the SCAR markers, species-specific PCR amplification and multiplex-PCR were carried out using the single species-specific primer pairs and combinations of them, respectively. Results : We used RAPD analysis of four Rheum plant species to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed two SCAR markers that amplified 314 bp and 390 bp DNA fragments in only R. undulatum but not in R. officinale, R. palmatum, R. tanguticum and R. undulatum, for distinguishing Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. Conclusions : These genetic markers can be used for the efficient discrimination of plants species and commercial herbal medicines between Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma, to ultimately prevent indiscriminate distribution and prescription of these herbal medicines.

• Korean Journal of Plant Resources
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• v.20 no.1
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• pp.79-85
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• 2007

The species in genus Saxifiraga distributed in circumpolar arctic are taxonomically difficult to study. RAPD analyses were performed to compare the genetic diversity of the 16 Saxifrages originated from the Norwegian Arctic Svalbard and Korea. The 12 accessions of URP primers were tested and 4 of which showed polymorphism were selected. Total 79 (44.8%) DNA bands were scored and analyzed by UPGMA cluster analysis. The results indicated that all of the 9 Saxifraga species from Svalbard showed high genetic diversity than those from Korea. The Similarity matrix and cluster analyses indicated that the Saxifraga species from Svalbard and Korea can be divided into two different subgroups. RAPDs of the Saxifraga species of Korea showed higher homologous patterns than those of Arctic Saxifrage. Among the Saxifraga species, we found that the morphological similarity reflects the genetic similarity. The geographic distance, clonal reproduction, and environmental condition may contribute the high level of genetic diversity between Saxifraga species from the two isolated regions.

• Journal of Mushroom
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• v.13 no.3
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• pp.243-249
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• 2015

Agaricus bitorquis is an edible white mushroom of the genus that is cultivated at high temperature($25{\pm}1^{\circ}C$) unlike A. bisporus is cultivate at $16{\pm}2^{\circ}C$. Unlike Agaricus bisporus, an edible white A. bitorquis mushroom is cultivated at high temperature ($25{\pm}1^{\circ}C$). Most farmers cultivate this mushroom for a long cultivation period in Korea. For this reason, we made heterokayons to develop a new cultivar that generate fruitbodies for short cultivation period. Over one hundred SSIs(single spores isolates) were collected from selected A. bitorquis ASI1151 and ASI1349 strains. Seventy-three SSIs were germinated on CDA(compost dextrose agar) media after 20 days (minimum) or 83 days (maximum) incubation under different media condition. The mycelial growth rate of germinated SSIs was different. 9 homokaryons in ASI 1151 and 11 homokaryons in ASI 1349 from SSIs were selected by OPN-02 primer in RAPD analaysis. Also this primer was used to select heterokaryon that cross among each homokaryon with compatible locus. Therefore 44 compatible matings were confirmed of 99 crossed lines.

• Korean Journal of Plant Resources
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• v.18 no.3
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• pp.359-366
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• 2005

Effects of neutron beam irradiation on seed germination, growth and RAPD pattern of tobacco (Nicotiana tabacum L. cv.; N. plumbaginifolia) and rice (Orya sativa L. cv.) plants were estimated. Seed germination rate was not significantly changed by the neutron beam treatment in both tobacco and rice seeds. And there was no significant differance in growth of the plants by the neutron beam treatment. Interestingly, however, some of morphological changes, including leaf shape (about $36\%$), stem color and leaf color were observed in neutron beam treated tobacco plants. In addition, abnormal flower in petal was observed in the neutron beam treated plant. This results indicate that neutron beam is able to use as an effective mutagen in plant mutations. Scorable products from 20 primers were obtained by RAPD analysis in the leaves of the beam irradiated tobacco plants and most of the plants showed the similar band patterns.

• The Korean Journal of Malacology
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• v.24 no.1
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• pp.11-24
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• 2008

GDNA was isolated from the jedo venus clam (Protothaca jedoensis, Lischke) from Boryeong (jedo venus clam from Boryeong JVCB) and Wonsan (jedo venus clam from Wonsan; JVCW) located in the West Sea and the East Sea of Korean Peninsula, respectively and we performed clustering analyses, DNA polymorphisms and the populations genetic variations. In the present study, the seven decamer primer generated the one hundred and eleven major/minor specific bands in JVCB population and ninety four-specific bands in JVCW population. Seven primers generated the unique shared bands to each population of one hundred and seventy-six, on average of 25,1, in JVCB population from Boryeong and three hundred thirty, on average of 47,1, in JVCW population from Wonsan, respectively. The dendrogram obtained by the seven oligonucleotides primers, indicates two genetic clusters. Especially, two Protothaca between the individual WONSAN no. 12 and BORYEONG no. 10 showed the longest genetic distance (0.537) in comparison with other individuals used. Accordingly, RAPD analysis showed that the JVCB was a little more genetically diverse than the JVCW population. This result implies the genetic similarity owing to rearing in the same and/or similar circumstances or inbreeding within the JVCW population. So to speak, JVCB population may have high levels of genomic DNA variability owing to the introduction of the wild individuals from the other sites to sampling sites although it may be the geographically diverse distribution of this species. However, it was confirmed that it did not appear like that really in this study. We feel convinced that RAPD analysis discovered a significant genetic distance between two Protothaca population pairs (P<0.001). The existence of population discrimination and genetic diversity between two Protothaca populations was identified by RAPD analysis.

• The Korean Journal of Mycology
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• v.26 no.4 s.87
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• pp.466-477
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• 1998

Twenty-two Phellinus strains were characterized using colony morphologies and polymerase chain reaction (PCR) to divide into Phellinus linteus. There were some differences in mycelial growth and colony shapes among the strains when they were grown on various media such as PDA, MCM, MEA and YM. Phellinus linteus was slowly growing, formed golden-yellow colony, and produced blue pigment on PDA media. When the regions of internal transcribed spacer (ITS) were amplified from ribosomal RNA (rRNA) coding genes of P. igniarius and P. linteus strains by means of PCR, two types of band (700 bp and 800 bp) were appeared, respectively. For the amplified intergenic region I (IGRI), P. igniarius strains showed a different band among 500, 600, 700 and 800 bp according to the strains, whereas P. linteus strains did one specific band of 700 bp. By polymorphism analysis after digesting the amplified products with 6 different restriction enzymes, a band specific to P. linteus was generated when the products for ITS region were digested with HaeIII, suggesting that the enzyme digestion could provide effective method to distinguish between P. igniarius and P. linteus. And also, the analysis of genetic relationship showed that the genetic similarities were 89% and 95% in P. igniarius and P. linteus strains, respectively. Random amplification polymorphic DNA (RAPD) analysis using multiple primer sets and arbitrarily primed PCR (AP-PCR) with ITS3 primer could also result in a reproducible way to identify P. linteus strains.

• 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.

• The Korean Journal of Mycology
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• v.38 no.2
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• pp.105-111
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• 2010

Lentinus lepideus, known as train wrecker fungus, has been used for nutritional and medicinal purposes. Recently, commercial cultivation technique and a new cultivar of the mushroom were developed. To investigate the genetic diversity and phylogenetic relationship for identifying the mushroom strains and cultivar, one commercial and 13 strains of Lentinus lepideus from different geographical regions of Korea were analyzed by ITS regions of rDNA and RAPD of genomic DNA. Three strains of Lentinus edodes were also used for the analysis. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 173 to 179 bp and 203 to 205 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical with 156 base pairs. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into four clusters, while 3 strains of L. edodes was divided into a new cluster. Ten primers out of 20 arbitrary primers used in the RAPD-PCR efficiently amplified the genomic DNA. The numbers of amplified DNA bands varied with the primers and strains, with polymorphic DNA fragments in the range from 0.2 to 2.6 kb. The results showed that phylogenetic relationship among Korean strains of Lentnus lepideus is high, but genetic diversity is low.

• Korean Journal of Medicinal Crop Science
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• v.21 no.6
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• pp.444-454
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• 2013

The development of random amplified polymorphic DNA (RAPD) and expressed sequence tag-derived simple sequence repeats (EST-SSRs) provided a useful tool for investigating Korean ginseng genetic diversity. In this study, 18 polymorphic markers (7 RAPD and 11 EST-SSR) selected to assess the genetic diversity in 31 ginseng accessions (11 Korean ginseng cultivars and 20 breeding lines). In RAPD analysis, a total of 53 unique polymorphic bands were obtained from ginseng accessions and number of amplicons ranged from 4 to 11 with a mean of 7.5 bands. Pair-wise genetic similarity coefficient (Nei) among all pairs of ginseng accessions varied from 0.01 to 0.32, with a mean of 0.11. On the basis of the resulting data, the 31 ginseng accessions were grouped into six clusters. As a result of EST-SSR analysis, 11 EST-SSR markers detected polymorphisms among the 31 ginseng accessions and revealed 49 alleles with a mean of 4.45 alleles per primer. The polymorphism information content (PIC) value ranged from 0.06 to 0.31, with an average of 0.198. The 31 ginseng accessions were classified into five groups by cluster analysis based on Nei's genetic distances. Consequently, the results of ginseng-specific RAPD and EST-SSR markers may prove useful for the evaluation of genetic diversity and discrimination of Korean ginseng cultivars and breeding lines.

• Korean Journal of Plant Resources
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• v.18 no.2
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• pp.260-269
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• 2005

Taxonomic studies on morphological, principal component analysis (PCA), palynological, RAPD and PCR-RELP analysis were conducted to intraspecific relationships of Rubus oldhami. Three types of Rubus oldhami based on the flower characters such as petal length and number were used in this study. Among the 14 morphological characters, perianth length, calyx lobe length, apical leaflet shape and leaflet length were used to distinguish for each type. The pollen characters such as shape, aperture number, surface sculpture were showed very similar among three types. Eight primers out of 20 arbitrary primers were screened for three types, and were revealed 33 ($60\%$) polymorphic bands. The phonogram by RAPD data showed incongruent with morphological analysis. Even though ten restriction endonucleases produced 20 restriction sites, polymorphic bands were not observed. Based on the results, three types of Rubus oldhami divided well by morphological characters, but pollen and DNA data were not supported. Therefore, type 1 and 2 which different from type 3 by flower characters considered as a temporary hybrid or ecotype because of their similar habitats.