• Title/Summary/Keyword: polymorphic bands

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Early Identification of Putative Zygotic Seedlings in Citrus Crosses between 'Morita unshiu' (Citrus. unshiu Marc.) and 'Ponkan' (C. reticulata Blanco) Using RAPD and SRAP (RAPD와 SRAP 방법을 이용한 '성전온주'(C. unshiu Marc.)와 '병감'(C. reticulate Blanco) 교잡실생 식별)

  • Yun, Su-Hyun;Moon, Young-Sun;Jin, Seong-Beom;Kang, In-Kyu;Lee, Dong-Hoon
    • Journal of Life Science
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    • v.21 no.4
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    • pp.502-508
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    • 2011
  • The purpose of this study was to evaluate the methods of identifying zygotic seedlings of crosses between 'Morita unshiu' (Citrus. unshiu Marc.) and 'Ponkan' (C. reticulata Blanco). In order to investigate the frequency and position of zygotic seedlings and to determine the efficiency of zygotic seedling identification, random amplified polymorphic DNA (RAPD) and sequence-related amplified polymorphism (SRAP) were performed using UBC (9, 27, 229, 230, and 254) primers and F4/R27, F7/R14, F12/R10, and F44/R62 primer sets, respectively. A total of 37 putative zygotic seedlings out of 55 individuals were selected by RAPD and SRAP. The F7/R14 primer pair showed a screening ability of 45.5% (25/55), whereas the primer UBC27 revealed the highest efficiency of zygotic seedling identification (50.9%, 28/55). When both UBC27 and F7/R14 were properly used for selection of hybridized seedlings of 'Morita unshiu' (C. unshiu Marc.) and 'Ponkan' (C. reticulata Blanco), screening efficiency was increased to 60% (33/55) for putative zygotic seedlings. Thus, it is possible to select putative hybrid zygotic seedlings in an accurate and effective manner by RAPD and SRAP.

The Discrimination of Coisis Semen and Coisis lacrima-jobi Semen by the Random Amplified Polymorphic DNAs and Anatomical Characteristics (의이인과 염주의 RAPD분석 및 해부학적 특징에 의한 감별)

  • Lee, Mi-Young;Im, Seung-Hi;Kim, Ho-Kyoung;Han, Keong-Sik;Choi, Yong-Hyu;Ju, Young-Seung;Oh, Seung-Eun;Ko, Byoung-Seob
    • 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.

The classification and comparison of genetic diversity of genus Malus using RAPD (RAPD를 이용한 능금속 식물종의 계통관계와 유전적 다양성)

  • Huh, Man-Kyu
    • Journal of Life Science
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    • v.17 no.6 s.86
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    • pp.756-761
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    • 2007
  • Cenus Malus is a long-lived woody species primarily distributed throughout Asia. Many species of this genus are regarded as agriculturally and ecologically important. The phynetics and genetic diversity among eight species of genus Malus were reconstructed using the random amplified polymorphic DNA (RAPD) markers. In a simple measure of intraspecies variability by the percentage of polymorphic bands, the M. micromalus exhibited the lowest variation (34.7%). The M. pumila showed the highest (50.0%). Mean number of alleles per locus (A) ranged from 1.347 to 1.500 with a mean of 1.437. The phenotypic frequency of each band was calculated and used in estimating genetic diversify (H) within species. The mean of H was 0.190 across species, varying from 0.155 to 0.220. In particular, two cultivated species, M. pumila and M. asiatica, had high expected diversity, 0.314 and 0.307, respectively. On a per locus basis, the proportion of total genetic variation due to differences among species ranged from 0.388 to 0.472 with a mean of 0.423, indicating that 42.3% of the total variation was found among species. The phylogenetic tree showed three distinct elates. One includes M. sieversii, M. pumila, and M. asiatica. Another includes three M. baccata taxa. The other includes M. sieboldii, M. floribunsa, and M. micromalus. One variety and one form of M. sieboldii were well separated each other. RAPD markers are useful in germ-plasm classification of genus Malus and evolutionary studies.

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.

Discrimination of Potato Varieties by Random Amplified Polymorphic DNA Analysis (RAPD에 의한 감자 품종의 구분)

  • Seo, Hyo Won;Yi, Jung Yoon;Cho, Hyun Mook;Park, Young Eun;Oh, Seung Eun
    • Horticultural Science & Technology
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    • v.19 no.1
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    • pp.29-33
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    • 2001
  • This study was carried out to discriminate potato cultivars and breeding lines by specific molecular markers using random amplified polymorphic DNA (RAPD) analysis. The genotypes of potatoes used for analysis were eight cultivars and five breeding lines. Some of those show much phenotypic resemblances among them because 'Jopung', 'Daekwan70', 'Gawon', and 'Daekwan72' have immediate parental relationship with 'Superior', 'Irish Cobbler', 'Namsuh', and 'Atlantic', respectively. So, there are many difficulties to distinguish the varieties by the morphological characteristics. Three URP primers, URP2, URP4, and URP8 were selected for promising primers to discriminate potato genotypes or cultivars. The three URP primers were shown very high reproducibility because of the relatively high annealing temperature and long primer size. Although the results of similarity analyses did not always reflect the genetic relationship between potato varieties, the reproducible pattern of amplified DNA bands by URP primers showed possibility for molecular markers for discrimination of potato genotype or cultivar.

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Genetic variations and relationships of Phragmites japonica and P. communis according to water environment change (수환경변화에 따른 갈대와 달뿌리풀의 유전적 변이 및 유연관계)

  • Kim, Yong-Hyun;Kim, Joo-Hwan
    • Korean Journal of Plant Resources
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    • v.22 no.2
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    • pp.152-158
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    • 2009
  • We performed RAPD analysis to investigate the inter-/intraspecific relationships and regional variations of Phragmites japonica and P. communis according to the environmental change. One hundred and fourty nine genetic effective polymorphic bands between 300 bp and 1,900 bp were marked from RAPD PCR with nine oligoprimers. From the RAPD analysis by Nei-Li's genetic distance, the dissimilarity indices among the populations of Phragmites japonica were relatively low from 0.012 to 0.061, and Phragmites communis were also low from 0.033 to 0.095. It showed the close genetic relationships among the same species populations, and both species were distinctly independent with relatively high level of dissimilarity indices (0.043 - 0.132). The obvious genetic markers to distinguish two species were confirmed and those profiles were suggested. From the UPGMA phenogram by RAPD analysis, both species showed the water environment related cluster patterns by distributional regions. RAPD analysis was useful to delimit two species taxonomically and to investigate the genetic relationships among inter-/intraspecific populations.

Analysis of Genetic Relationship of Pear (Pyrus spp.) Germplasms Using AFLP Markers (AFLP 표지를 이용한 배 유전자원의 유연관계 분석)

  • Cho, Kang-Hee;Shin, Il Sheob;Kim, Hyun Ran;Kim, Jeong-Hee;Heo, Seong;Yoo, Ki Yeol
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.444-450
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    • 2009
  • Amplified fragment length polymorphism (AFLP) marker was utilized for evaluation of genetic diversity of 60 pear germplasms. Twenty selective AFLP primer pairs generated a total of 522 polymorphic amplification products. From UPGMA (unweighted pair-group method arithmetic average) cluster analysis by using polymorphic bands, the pear germplasms were divided into four clusters by similarity index of 0.691. The first cluster (I) included European pears belonging to Pyrus communis and wild species such as P. nivalis and P. cordata. The second cluster (II) included Ussurian pea pears belonging to P. betulaefolia and P. fauriei. The third cluster (III) included pea pears belonging to P. calleryana and P. koehnei. Most of germplasms belonging to P. pyrifolia and P. ussuriensis, and interspecific hybrids were included in the fourth (IV) cluster. Therefore pear germplasms originated from East Asia were closely related to P. pyrifolia and P. ussuriensis. Similarity values among the tested pear germplasms ranged from 0.584 to 0.879, and the average similarity value was 0.686.

Polymorphism analysis of tri- and tetranucleotide repeat microsatellite markers in Hanwoo cattle

  • Shil Jin;Jeong Il Won;Hyoun Ju Kim;Byoungho Park;Sung Woo Kim;Ui Hyung Kim;Sung-Sik Kang;Hyun-Jeong Lee;Sung Jin Moon;Myung Sun Park;Yong Teak Sim;Sun Sik Jang;Nam Young Kim
    • Journal of Animal Science and Technology
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    • v.66 no.4
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    • pp.717-725
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    • 2024
  • The Hanwoo traceability system currently utilizes 11 dinucleotide repeat microsatellite (MS) markers. However, dinucleotide repeat markers are known to have a high incidence of polymerase chain reaction (PCR) artifacts, such as stutter bands, which can complicate the accurate reading of alleles. In this study, we examined the polymorphisms of the 11 dinucleotide repeat MS markers currently employed in traceability systems. Additionally, we explored four trinucleotide repeat MS markers and one tetranucleotide repeat MS marker in a sample of 1,106 Hanwoo cattle. We also assessed the potential utility of the tri- and tetranucleotide repeat MS markers. The polymorphic information content (PIC) of the five tri- and tetranucleotide repeat markers ranged from 0.663 to 0.767 (mean: 0.722), sufficiently polymorphic and slightly higher than the mean (0.716) of the current 11 dinucleotide repeat markers. Using all 16 markers, the mean PIC was 0.718. The estimated probability of identity (PI) was 3.13 × 10-12 using the 11 dinucleotide repeat markers, 7.03 × 10-6 using the five tri- and tetranucleotide repeat markers, and 2.39 × 10-17 using all 16 markers; the respective PIhalf-sibs values were 2.69 × 10-9, 1.29 × 10-4, and 3.42 × 10-13; and the respective PIsibs values were 3.89 × 10-5, 9.6 × 10-3, and 3.69 × 10-7. The probability of exclusion1 (PE1) was 0.999864 for the 11 dinucleotide repeat markers, 0.981141 for five of the tri- and tetranucleotide repeat markers, and > 0.99 for all 16 markers; the respective PE2 values were 0.994632, 0.901369, and > 0.99; and the respective PE3 values were 0.998702, > 0.99, and > 0.99. The five investigated triand tetranucleotide repeat MS markers can be used in combination with the 11 existing MS markers to improve the accuracy of individual identification and paternity testing in Hanwoo.

Detection of Pseudomonas tolaasii Causing Brown Blotch Disease of Mushroom with Species-specific DNA Probe (종 특이 DNA probe를 이용한 버섯 세균성 갈반병 병원균(Pseudomonas tolaasii)의 검출)

  • Kwon, Soon-Wo;Go, Seung-Joo;Cheun, Meung-Sook;Kang, Hee-Wan;Oh, Se-Jong;Chang, Who-Bong;Ryu, Jin-Chang
    • The Korean Journal of Mycology
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    • v.27 no.2 s.89
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    • pp.132-137
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    • 1999
  • This study was carried out to develop the molecular marker for the detection of Pseudomonas tolaasii, a causative agent of bacterial brown blotch disease of oyster mushroom (Pleurotus ostreatus). When several primers designed from repetitive sequences and pectin lyase genes of bacteria were used to produce DNA polymorphism from different Pseudomonas spp. isolated from edible mushrooms, PEU1 primer derived from pectin lyase gene produced polymorphic bands differentiating P. tolaasii strains from other Pseudomonas species. Two bands, 1.0kb and 0.4kb, found commonly in 6 isolates of P. tolaasii were cloned into pGEM-T vector which were designated as pPTOP1 and pPTOP2, respectively, to use as probe. The 0.4 kb insert of pPTOP2 hybridized to only 6 isolates of P. tolaasii, but did not to the other Pseudomonas species. As few as $1.5{\times}10^3$ colony forming unit (cfu) of P. tolaasii could be detected by dot blot hybridization with the cloned 0.4kb DNA in pPTOP2.

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Pholiota adiposa and its Related Species Collected from the Wild Forestry (야생에서 채집된 검은비늘버섯(Pholiota adiposa)균에 관한 연구)

  • Lee, Sang-Sun;Kim, Mi-Hye;Chang, Hu-Bong;Shin, Chun-Sik;Lee, Min-Woong
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.574-582
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    • 1998
  • Five basidiocarps of Pholiota species have been collected from the areas of BubJu Temple for last two years, and identified to those of P. adiposa or Pholiota species. The taxonomy of these basidiocarps with the morphological aspects was compared with the analysis obtained from the polymorphisms of PCR-RAPD bands made after reacted with the random primers. The polymorphic variations were observed within the species of the basidiocarps, but not between genomic DNA's of the mycelia obtained and the basidiocarps. Several different bands made from the primers (28 and 36) in PCR-RAPD reactions were identified within the genus of Pholiota and speculated to be specific for the individual basidiocarp of P. adiposa collected. The primers employed here were considered to be very useful for distinguishing the individual isolates or basidiocarps collected from the fields. Also, the basidiospores were obtained from the sporeprints of the above basidiocarps as a simple agar and confirmed with observations of clamp connection under microscopes. The matings of them indicated the 'tetrapolar' type, being different from the 'bipolar' type reported by Japanese basidiocarps of P. adiposa or P. nameko. Based on our work, the edible fungi collected were speculated to be a new breeding resource for those of Pholiota commercialized in Japan.

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