• Title/Summary/Keyword: RAPD technique

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Interactions of Newly Isolated Orchid Mycorrhizal Fungi with Korean Cymbidium kanran Hybrid 'Chungsu'

  • Lee, Jun-Ki;Lee, Sang-Sun;Eom, Ahn-Heum;Paek, Kee-Yoeup
    • Mycobiology
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    • v.31 no.3
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    • pp.151-156
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    • 2003
  • Two fungal isolates obtained from roots of Cymbidium goeriingii in Jeju island were confirmed to be symbiotic with orchid plantlets, and were compared with other orchid mycorrhizal(OM) fungi previously isolated. The two isolates differed in their peloton structures formed in the roots of Cymbidium kanran hybrid 'Chungsu' and in responses of orchid plant. These two isolates differed from the additionally tested OM fungi in some features, and from root damaging species of Rhizoctonia and Fusarium as based on cluster analysis after PCR-RAPD with the primers, Bioneer-28 and OPO-2. With this simple and fast technique, it was possible to distinguish OM fungi from the plant root pathogenic fungi based on calculation of their polymorphic bands. This technique can therefore be helpful to distinguish the OM fungi from the root pathogens. Particularly, the new isolates are considered as new resource of symbiotic fungi for horticultural industries.

Random Amplified Polymorphic DNA (RAPD) Identification of Genetic Variation in Chlorella species

  • CHO Jung Jong;KIM Yong-Tae;HUR Sung Bum;KIM Young Tae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.6
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    • pp.761-769
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    • 1996
  • The random amplified polymorphic DNA (RAPD) technique was used to characterize 18 reference strains of microalgae, mostly Chlorella species, collected from various localities around Korea peninsular. Eighteen strains consist of four genera of the family marine Chlorella from 12 samples, two genera of fresh water Chlorella from three samples, and three genera on Nannochloris. Twenty 10-mer anonymous primers were screened for amplification of genomic DNA extracted from samples using the CTAB extraction method. Nineteen of these oligonucleotide primers were positive or band producing. Three of 20 random primers (OPA 10, OPA 12, and OPA 18) resulted in both clear band and a high degree of reproducibility and showed some potential to be used to discriminate individual samples of both genetically hetero-and homogeneous populations, in determining phylogenetic relationships between species within a genus and developing individual fingerprints for each samples.

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Analysis of Genetic Relationship by RAPD Technique for Codonopsis lanceolata Trauty Collected from the Baekdoo Mountain and Korea (백두산지역과 국내 더덕 수집종의 RAPD에 의한 유연관계 분석)

  • Doo, Hong-Soo;Ryu, Jeom-Ho;Lee, Kang-Soo;Li, Hu Lin;Liu, Xian Hu
    • 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.

Development and Application of PCR-based Markers for the Discrimination of Bang-Poong and Related Species (방풍류의 감별을 위한 분자마커의 탐색과 활용)

  • Hong, Seong-Mi;Lee, Mi-Young;Koh, Jae-Chul;Ko, Byoung-Soeb
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.1-6
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    • 2004
  • Bang-Poong and related species are an important herbal medicine. However, it is difficult to determine the commercial dry material through anatomical and chemotaxonomical characteristics. Here, we used a PCR-based technique for an accurate discrimination of Bang-Poong and related species. With the RAPD primers, 215 RAPDSs(random amplified polymorphic DNAs) were obtained, and 98% of them showed polymorphic patterns. RAPDs from the four primers were appropriate for the discrimination of S. divaricata $(T_{URCZ{\cdot}})\;S_{CHISKIN}$, those from the six primers for P. japonicum $T_{HUNBERG}$, those from the four primers for P. terebinthaceum $F_{ISHER}$, and those from the six primers for G. littoralis Fr. $S_{CHMIDT}$. The specific bands from the primer 425 were obtained and used to develop SCAR (sequence characterized amplified region) markers, based on the sequence information of the RAPD markers. The SCAR primers generated a 215 bp fragment specific to Peucedanum terebinthaceum $F_{ISHER}$, and a 177 bp and a 300 bp fragment specific to G. littoralis Fr. $S_{CHMIDT}$. As a result, the three SCAR markers were able to discriminate from two Bang-Poong related species.

Genetic Diversity and Population Structure of Brassica juncea by Random Amplified Polymorphic DNA (RAPD) (RAPD를 이용한 겨자의 유전적 다양성과 집단구조)

  • Oh, Yung-Hee;Moon, Sung-Gi;Chae, Yang-Hee;Hong, Hwa-Jin;Cho, Min-Cheol;Park, So-Hye;Huh, Man-Kyu
    • Journal of Life Science
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    • v.20 no.10
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    • pp.1538-1543
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    • 2010
  • This study was conducted to examine the genetic diversity and population structure of 17 Brassica juncea populations in Korea. The technique of random amplified polymorphic DNA (RAPD) produced 60 polymorphic loci and 18 monomorphic loci. In a simple measure of intraspecies variability by the percentage of polymorphic bands, the Jindo population of Cheonnam showed the highest (29.5%). The cultivar exhibited the lowest variation (12.8%). Mean number of alleles per locus (A) and the effective number of alleles per locus ($A_E$) were 1.221 and 1.167, respectively. As the typical populations of this species were small, isolated, and patchily distributed in their natural populations, they maintained a low level of genetic diversity of fourteen primers. On a per locus basis, total genetic diversity values ($H_T$) and interlocus variation in the within-population genetic diversity ($H_S$) were 0.347 and 0.141, respectively. On a per-locus basis, the proportion of total genetic variation due to differences among populations ($G_{ST}$) was 0.589. This indicated that about 58.9% of the total variation was among populations. The estimate of gene flow, based on $G_{ST}$, was very low among Korean populations of B. juncea ($N_m$=0.617). These results suggest that the geological distance dispersal of wild B. juncea is the best event. RAPD markers are very effective in classifying natural population levels of B. juncea in Korea.

RAPD Identification of Genetic Variation in Ulvales Seaweed (RAPD기법을 이용한 갈파래목 해조류의 유전 변이 분석)

  • CHO Yong-Chul;PARK Ji Won;JIN Hyung-Joo;NAM Bo-Hye;SOHN Chul Hyun;HONG Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.3
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    • pp.388-392
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    • 1997
  • The random amplified polymorphic DNAs (RAPD) technique was used to characterize seven isolates of the green seaweed Ulvales collected from Songjeng, Haeundae, Jumunjin, Dadaepo and Wando in Korea. Total DNA was extracted by the LiCl extraction method from thalli of green seaweed. The extracted DNA (3 ng) in $25{\mu}\ell$ reaction volume was amplified by 45 cycles of the polymerase chain reaction with arbitrary primers. Thirty-four primers resulted in 1227 PCR products ranged 240 bp to 1.5 kb of both conserved and polymorphic bands. Genetic similarities of the seven isolates calculated by Jaccard's equation were ranged from $7\%\;to\;36\%$. Monostroma nitidum (Wando) was shown to be most distantly related with the others based on genetic similarity and did not produce the amplified band of 630 bp, common in Ulvales using primer OPB-01 (CATCCCCTG).

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Differentiation of indigenous balloon flower (Platycodon grandiflorum DC.) germ lines in South Korea by using RAPD analyses (RAPD분석 기술을 이용한 토종도라지의 기원 분석)

  • Kim, Tae-Won;Lee, Soo-Jin;Kim, Man-Bae;Park, Chun-Geon;Shin, Yong-Wook;Cho, Young-Son;Lee, Shin-Woo
    • Journal of Plant Biotechnology
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    • v.41 no.1
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    • pp.19-25
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    • 2014
  • The total production volume has been sharply increased from year 2008 in Gyeongnam province, South Korea by the policy of preservation and promotion of indigenous balloon flower germ lines. In an attempt to assist the Gyeongnam province's policy, in this study, we tried to establish a technique to differentiate the indigenous balloon flower germ lines with those collected within South Korea and China. Our preliminary results indicated that RAPD analyses with five different primers exhibited high frequency of polymorphic DNA bands up to 76.9% and phylogenetic tree indicated that some of the indigenous lines can be easily differentiated with others. However, it was suggested that more advanced techniques such as single nucleotide polymorphic markers need to be developed in particular, by using extra-chromosomal DNA.

Assessment of Genetic Diversity of Hedera spp. Using RAPD Marker Technique (RAPD Marker를 이용한 Hedera속 식물의 다양성 조사)

  • Jung, Mi Soon;Joung, Youn Hwa;Lee, Ja Hyun;Choi, Jeong Keun;Kim, Kwang Soo;Han, Tae Ho
    • FLOWER RESEARCH JOURNAL
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    • v.16 no.1
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    • pp.28-35
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    • 2008
  • Eleven accessions of Hedera helix, three accessions of Hedera rhombea, one accession of Fatshedera lizei, and one accession of Fatsia japonica were collected and their genetic diversity was measured by using 10 RAPD primers. Approximately ninety seven percentage of polymorphism was detected, because broad germplasm, three genus, was used. Total 97 bands were scored and a dendrogram was constructed by using an UPGMA method. Accessions belonging to Hedera helix tightly clustered in one group: eight accessions showed extremely narrow genetic differences and the other three accessions also showed significant similarity. Despite of their genetic similarity, they showed morphological variations. The morphological variation with a narrow genetic differences indicated that the ivy cultivars have been indeed developed from a mutation breeding program. Genetically most unrelated Fatsia japonica showed a genetic distance of 0.63 on the average between other species. An accession from Fatshedera lizei developed by crossing between two genus, Hedera helix and Fatsia japonica, was allocated together with accessions from Hedera rhombea.

Analysis of Genetic Relationships of Grapevine Cultivars (Vitis ssp.) in Korea Using RAPD Markers (RAPD를 이용한 한국 포도 품종의 계통유연관계 분석)

  • Yoo, Ki Yeol;Cho, Kang-Hee;Shin, Il-Sheob;Kim, Jeong Hee;Heo, Seong;Noh, Jung Ho;Kim, Hyun Ran
    • 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").