• Title/Summary/Keyword: RAPD primer analysis

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Molecular Genetic Classification of Hypsizigus marmoreus and Development of Strain-specific DNA Markers (느티만가닥버섯의 분자유전학적 분류 및 품종특이적 DNA 마커 탐색)

  • Lim, Yun-Jeong;Lee, Chang-Yun;Park, Jeong-Eun;Kim, Sang-Woo;Lee, Hyun-Sook;Ro, Hyeon-Su
    • The Korean Journal of Mycology
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    • v.38 no.1
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    • pp.34-39
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    • 2010
  • We have attempted to verify 30 strains of Hypsizigus marmoreus from various mushroom stocks in Korea using random amplified polymorphic DNA (RAPD) methodology. Chromosomal DNAs of them were extracted and subjected to PCR analyses with 3 random primers. Each PCR produced approximately 30 distinct PCR bands with the size from 200 bp to 3000 bp. A dendrogram was acquired using the unweighted pair-group method with arithmetic average (UPGMA) clustering methodology on the basis of the DNA band pattern. The analysis revealed that 30 strains of H. marmoreus were clustered into two distinct clusters. Cluster 1 contained 3 subgroups while the cluster 2 consisted of rather diverse strains. Interestingly, Hm3-10, a wild strain collected from Deog-Yu mountain, was not included in either clusters, indicative of uniqueness of this strain. We nextly attempted to develop strain-specific DNA markers to verify a specific strain. A unique band in the RAPD gel lane of Hm0-4 was extracted and its sequence was determined. PCR with a primer set from the determined sequence revealed that the primer set gave a 250 bp DNA band only for Hm0-4, indicating that this approach works well for the strain-specific identification of H. marmoreus.

Analysis of Genetic Relationships of Korean Native Polygonatum spp. (우리 나라 자생 둥굴레속 식물의 유연관계 분석)

  • Jang, Kye-Hyun;Song, Geun-Woo;Chung, Jong-Il;Kang, Jin-Ho
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.3
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    • pp.214-218
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    • 2004
  • Germplasms having valuable characters are increasjngly important for modern breeding programs. This study was conducted to obtain the basic data for effective use of genetic resources of Polygonatum. The relationship of seven Polygonatum species collected widely in Korea was analyzed by RAPD markers. Total number of alleles amplified by nineteen random primers were 114 to 157, and variation in number of alleles was also diverse among seven species examined. The seven species were divided into two groups; one was of Polygonatum stenophyllum and Polygonatum humile. the other was of Polygonatum inflatum, Polygonatum lasianthum, Polygonatum odoratum var. pluriflorum (1), (2), and for variegatum.

Genetic Variability of Pleurotus ostreatus Monospore Isolates by Random Amplified Polymorphic DNA Analysis (DNA 다형성 분석에 의한 느타리버섯 단포자 분리주의 유전적 변이성)

  • Song, Yeong-Jae;Jeong, Mi-Jeong;Kim, Beom-Gi;Rho, Yeong-Deok;Ryu, Jin-Chang;Yoo, Young-Bok
    • The Korean Journal of Mycology
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    • v.24 no.3 s.78
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    • pp.186-205
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    • 1996
  • This study was carried out to obtain data concerning the genetic variability of Pleurotus ostreatus. Monospores of P. ostreatus were isolated and cultured to estimate differences in the rate of mycelial growth and genetic similarity among the isolates. Although the overall growth rates were slow compared to their parental dikaryon except 2-MI 4, significant differences in the rate of mycelial growth were observed among isolates. Random amplified polymorphic DNA (RAPD) analysis using twenty six random primers showed 345 polymorphic DNA bands from 35 monospore isolates and their parental dikaryon. DNA bands ranged from 0.1 to 4.0 Kb in size. Most various polymorphic DNA bands within monospore isolates were obtained when we used J (OPA-01) or W (OPB-04). The 36-MI 103 showed totally different band patterns from those of the others. RAPD analysis revealed that there is approximatly 75% genetic similarity between monospore isolates and their parental dikaryon except 36-MI 103, which showed only 49% genetic similarity. In addition, genetic similarity degrees were classified into four groups: I (parental dikaryon), II (fast growing type), III (moderate growing type), and IV (slow growing type). However, there is no correlation between mating type and mycelial growth rates.

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Species Identification of the Tropical Abalone (Haliotis asinina, Haliotis ovina, and Haliotis varia) in Thailand Using RAPD and SCAR Markers

  • Klinbunga, Sirawut;Amparyup, Piti;Leelatanawit, Rungnapa;Tassanakajon, Anchalee;Hirono, Ikuo;Aoki, Takashi;Jarayabhand, Padermsak;Menasveta, Piamsak
    • BMB Reports
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    • v.37 no.2
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    • pp.213-222
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    • 2004
  • A randomly amplified polymorphic DNA (RAPD) analysis was used to identify the species- and population-specific markers of abalone; Haliotis asinina, H. ovina, and H. varia in Thai waters. Fifteen species-specific and six population-specific RAPD markers were identified. In addition, an 1650 bp band (UBC195) that was restricted to H. ovina from the Gulf of Thailand (east) was also found. All of the specific RAPD markers were cloned and sequenced. Twenty pairs of primers were designed and specificity-tested (N = 12 and 4 for target and non-target species, respectively). Seven primer pairs (CUHA1, 2, 4, 11, 12, 13, and 14) were specifically amplified by H. asinina DNA, whereas a single pair of primers showed specificity with H. ovina (CUHO3) and H. varia (CUHV1), respectively. Four primer pairs, including CUHA2, CUHA12, CUHO3, and CUHV1, were further examined against 216 individuals of abalone (N = 111, 73, and 32, respectively). Results indicated the species-specific nature of all of them, except CUHO3, with the sensitivity of detection of 100 pg and 20 pg of the target DNA template for CUHA2 and CUHA12 and CUHV1, respectively. The species-origin of the frozen, ethanol-preserved, dried, and boiled H. asinina specimens could also be successfully identified by CUHA2.

Relatedness Among Indiginous Members of Sclerotinia sclerotiorum by Mycelial Compatibility and RAPD Analysis in the Jordan Valley

  • Osofee, H.;Hameed, K.M.;Mahasneh, A.
    • The Plant Pathology Journal
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    • v.21 no.2
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    • pp.106-110
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    • 2005
  • Sclerotinia sclerotiorum attacks most of the vegetable crops in the Jordan valley. Twenty-five samples/isolates were obtained in a complete coverage of that region. They were characterized for their mycelium incompatibility, and specific gene amplified using the primer SSREV/SSFWD. All isolates gave similar single band around 278 bp. Thirteen isolates were completely incompatible with the other 12 ones. The latter ones fell into four subgroups of mycelium incompatibility. RAPD analysis using three primers (OPA-2, OPA-10, and OPA-18) clustered the 25 isolates into subgroups in agreement with their morphological separation, indicating close correlation between amplified gene(s) and the gene(s) of incompatibility. All highly virulent isolates were among the group of 13, indicating a well established genomic type pathogen in this region.

Development of Narrow Leaf Type Cultivar "Konhee" in Zoysiagrass (세엽 한국잔디류 신품종 '건희(Konhee)' 육성)

  • 김두환;이재필;김종빈;모숙연
    • Asian Journal of Turfgrass Science
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    • v.13 no.3
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    • pp.147-151
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    • 1999
  • "Konhee" zoysiagrass[Zoysia matrella (L) Merr.](Patent registration no. 2000-723), a vegetative cultivar, was developed by the Dept. of Horticultural Science, Konkuk University, Seoul. "Konhee" was selected from the cross, ZKV6$\times$ZKV10, in 1997 and F1 seed were produced in the greenhouse in 1996. "Konhee" is superior to the other fine leaf zoysiagrass lines in many traits such as erect type, plant height($8.5\pm$2.0cm), short 3rd stolon length($3.4\pm$0.5cm), dark green, fine leaf($2.3\pm$0.2mm), low first sheath height($0.9\pm$0.2cm), rapid establishment and recoverage, many stolon number, and high shoot density. When "Konhee" was compared to the five other zoysisagrass lines at the DNA level using 35 PCR primers, it had the specific bands with primer No. 740, 544, 765, 772 by RAPD analysis.No. 740, 544, 765, 772 by RAPD analysis.

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Development of RAPD-SCAR Molecular Marker Related to Seed-hair Characteristic in Carrot (당근(Daucus carota var. sativa) 종자모 형질 관련 RAPD-SCAR 분자표지 개발)

  • Shim, Eun-Jo;Park, Sung-Kwan;Oh, Gyu-Dong;Jun, Sang-Jin;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.756-763
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    • 2013
  • Mechanical hair removal of carrot seed causes seed injuries and suppresses the germination in carrot cultivation. This study was performed to develop molecular markers for breeding high quality cultivars with short-hair seed. To meet this objective, random amplified polymorphic DNA (RAPD)-sequence characterized amplified region (SCAR) markers specifically linked to seed-hair characteristic were identified using CT-SMR 616 OP 389-1 line with short-haired seed and CT-SMR 616 OP 616-33 line with long-haired seed, bred by self-pollination for 6 years from 2008 to 2013, as parents. After seed hair lengths of these lines were analyzed using microscope, next generations were advanced and compared with the molecular markers polymorphism. From RAPD analysis using fixed lines in 2011, twelve RAPD primers showing polymorphic bands specific between the two lines were identified from 80 random primers. To develop RAPD-SACR marker, SCAR primers were designed based on sequence analysis of these specific RAPD bands and more than three combinations of primers were tested. As a result, it was found that the $SCA2_{1.2}$ amplified single polymorphic band from short-haired seed line. To confirm this result, $SCA2_{1.2}$ marker was retested by applying to the 2012 and 2013 progenies. Finally, it was concluded that the developed $SCA2_{1.2}$ marker distinguished short-haired line from long-haired seed line. Therefore, SCAR marker, $SCA2_{1.2}$ is expected to be utilized for breeding of the short-haired seed cultivars.

Genetic Relationship of Genus Rosa Germplasm and Genetic Diversity of Rosa rugosa in Korea (장미속 유전자원의 유전적 관계와 국내 해당화의 유전적 다양성)

  • Joung, Youn-Hwa;Kim, Seung-Tae;Kim, Gi-Jun;Lee, Ja-Hyun;Gi, Gwang-Yeon;Han, Tae-Ho
    • Horticultural Science & Technology
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    • v.28 no.6
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    • pp.1003-1013
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    • 2010
  • Fifty-nine accessions of 23 species in genus Rosa were collected, and 15 accessions of Rosa rugosa were collected throughout 10 regions of Korea. Their genetic relationship was investigated by using morphological analysis and RAPD marker. The morphological analysis was measured for 7 quantitative and quantified for 4 qualitative traits. RAPD analysis obtained a total of 959 polymorphic bands by using twenty primers. Morphological analysis classified most according to the rose section system except for several species. The cluster analysis of genus Rosa based on RAPD data could identify the subgenus $Platyrhodon$ and $Eurosa$. The subgenus $Eurosa$ separated five sections; $Gallicanae$, $Cinnamomeae$, $Pimpinellifoliae$, $Synstylae$ and $Caninae$. Correlation analysis between morphological and RAPD analysis showed low significance ($r$ = 0.35). The accessions of R. rugosa belonged to the section $Cinnamomeae$ clustered into three groups at genetic distance ranging from 0.28 on the base of RAPD analysis. In conclusion, the genetic relationship of the genus Rosa was consistent to the previously reported rose section system, and domestic collections of $R.$ $rugosa$ were separated from 3 groups on the base of RAPD marker.

Analysis of Genetic Relationships in Hypericum erectum Thunb. by RAPD (RAPD 방법을 이용한 고추나물의 유연관계분석)

  • Kim, Sun-Hee;Kim, Eung-Sik;Kim, Sung-Ho;Ahn, Jun-Cheul;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.4
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    • pp.141-145
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    • 2005
  • The genetic relationships of Hypericum erectum, H. ascyron and H. perforatum Thunb. by RAPD using total 46 primers were analysed 30 primers among primers tested showed the amplification band in all. The amplified DNA fragment ranged from 0.25 to 6.6 kb in size. The 411 bands (34.4%) among 1194 bands derived from 30 primers were polymorphic, and 13.7 bands per primer were observed. The phenograms for six analyzed individuals by RAPD markers were not matched well with those of the result by morphological characters. They were clustered monophyletic at the similarity coefficient value ranged from 0.24 to 0.96.

Construction of a Genetic Linkage Map in Radish(Raphanus sativus L.) Using RAPD Markers (RAPD 마커를 이용한 무의 유전자지도 작성)

  • Ahn, Choon-Hee;Choi, Su-Ryun;Lim, Yong-Pyo;Chung, Hae-Joon;Yae, Byeong-Woo;Yoon, Wha-Mo
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.151-159
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    • 2002
  • Genetic map and molecular marker have a great importance in improving and facilitating crop breeding program as well as in genome analysis and map-based cloning of genes representing desirable characters. This study aimed at developing RAPD markers and constructing a genetic linkage map using 82 BC$_1$F$_1$individuals originated from the cross between '835' and B$_2$in radish (Raphanus sativus L.). One of the parents for genetic linkage map construction, '835'(P$_1$) of egg type is susceptible to Fusarium wilt and have medium resistance to virus infection and the other parent, B$_2$(P$_2$) of round type, is susceptible to Fusarium wilt and virus, Screening of 394 RAPD primers in BC$_1$F$_1$) population resulted in selecting 128 polymorphic markers which displayed 1:1 segregation pattern. Two markers failed to display 1:1 segregation and showed the segregation ratio skewed to maternal genotype. Selected markers were categorized into 14 linkage group based on LOD score represented by MAPMAKER/EXP program. Five groups composed of single marker among them were excluded from the linkage map, and consequently, the remaining groups are well matched with the number of radish chromosome (n=9). The linkage map constructed with 128 markers covers 1,688.3 cM and the average distance between markers was 13.8 cM. For developing STS marker, we determined the partial nucleotide sequence of OPE10 marker at both ends and designed a oligonucleotide primer pair based on this sequence. STS PCR using the primer pair displayed a single, clear band of which segregation is perfectly matched with that of OPE10 marker. This implies that RAPD markers could readily convert into clear and reliable STS markers.