• 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.

• Weed & Turfgrass Science
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• v.2 no.2
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• pp.191-197
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• 2013

Two medium-leaf ecotypes (CY6069, CY6097) belonging to one species (Zoysia japonica) of Korean lawngrasses were selected in sod production fields in Jang Seong, Korea. They were reported to have distinct morphological and growth rate characteristics different from the preferred medium-leaf type zoysiagrass in Korea. This study was conducted to define further the genotypic difference at the molecular level and to develop DNA marker based on the specific DNA fragment. Polymorphic DNA fragments were first explored by using randomly amplified polymorphic DNA (RAPD) primers, which were then converted into PCR-based sequence characterized amplified region (SCAR) markers. The CY6069-specific primer set amplified about 550 bp successfully, while the CY6097 marker produced the expected 690 bp band, by which those markers were nominated by CY6069_550 and CY6069_690 SCARs, respectively. Together with the reported morphological and other phenotypic features, the SCAR markers confirmed in this study will be useful to identify those medium-leaf zoysiagrass genotypes when they are cultivated with other vegetatively propagated warm-season turfgrasses in sod farms.

• Parasites, Hosts and Diseases
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• v.33 no.4
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• pp.341-348
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• 1995

Genetic status of Acnnthamoebc sap. were tested on the basis of random amplified polymorphic DNA (RAPD) marker analysis. Four previously established Accnthcmoebn species, 4 Korean isolates of Acnnthamoeba sp., and one American isolate of Acanthcmoebc sp. were analyzed by RAPD-PCR using an arbitrary decamer primers. Amplification products were fractionated by agarose gel electrophoresis and slainrd by ethidium bromide . Eighteen primers produced DNA amplification profiles revealing clear differences among 4 species. Nine of them also produced DNA amplification profiles which included some isolate-specific amplification products. On the basis of amplified fragments by 18 primers, the pairwise similarity indices between A. culbensoni and other species (i.e. A. hntchetti, A. trinngularis, A. polyphaga) were 0.300, 0.308, and 0.313, respectively. Similarity index between A. hctchetti and A. triansulcris was 0.833. The mean similarity index among the 3 Korean isolates (YM-2, -3, -4) was 0.959 and 0.832 among them and 2 other species (A. hatchetti and A. triongulnris). The mean similarity index among YM-5 and other Korean isolates (YM-2, -3, -4) was 0.237. However, the similarity index between YM-5 and A. culbeksoni was 0.857, which suggests that YM-5 is genetically more similar to A. culbertsoni than other Korean isolates. Phonogram reconstructed by UPGMA method revealed that there are two groups: one group consists of A. hctchetti, A. tlonsulcns, and 3 Korean isolates (YM-2, -3, -4) , and the other group consists of A. cuLbensoni. A. polwphosc, HOV, and YM-5.

• Journal of Life Science
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• v.31 no.6
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• pp.537-542
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• 2021

Schisandra nigra Max., a dioecious plant native to Jeju Island in Korea, is cultivated on a small scale for fruit production. As fruit-producing female individuals are generally considered to be more valuable than male, early identification of plant sex at the seedling stage is important. In this study, a sequence-characterized amplified region (SCAR) marker associated with a female-specific region in the genome of S. nigra was investigated. Of 120 randomly amplified polymorphic DNA (RAPD) primers, one primer (OPB-03) consistently amplified a 749 bp band in female plants. The female-specific PCR product was isolated and cloned, and the nucleotide sequences were then determined. Southern hybridization performed using the female-specific fragment as a probe produced positive signals only in genomic DNA from the female plants. This result revealed that the 749 bp segment of DNA was present in the genome of female plants but absent in the genome of male plants. A SCAR primer pair was designed based on the RAPD marker to amplify a 436 bp fragment in the genomic DNA of female plants. This primer pair amplified the expected size of DNA fragment in female plants and four monoecious individuals collected from a natural population. The SCAR marker identified in this study can be used to distinguish female-flowering individuals at the seedling stage.

• Horticultural Science & Technology
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• v.31 no.6
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• pp.798-806
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• 2013

Precise, fast, and cost-effective identification of crop cultivars is essential for plant breeder's rights. Traditional methods for identification of persimmon cultivars are based on the evaluation of sets of morphological characteristics. However, it is difficult to distinguish closely related cultivars using only morphological traits. This study was conducted to develop DNA markers for identification of the 32 persimmon cultivars in Korea and Japan. A total of 309 randomly amplified polymorphic DNA (RAPD) markers were identified using 40 different random primers. Various number of polymorphic bands ranged from 4 (OPP-08) to 14 (UBC159) were detected with an average of 7.7. Resulting 57 RAPD fragments were selected, and their sequences were determined for developing sequence characterized amplified region (SCAR) markers. As a result, 15 of 57 RAPD fragments were successfully converted to SCAR markers. Single polymorphic bands of the same size as or smaller than the RAPD fragments were amplified depending on SCAR markers. Among these markers, a combination of eight SCAR markers (PS225_200, PSN05_420, PSF13_523, PSN11_540, PS372_567, PS485_569, PSP08_635, and PS631_735) provided sufficient polymorphisms to identify 32 persimmon cultivars. These newly developed markers will be a fast and reliable tool to identify persimmon cultivars.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2002.11b
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• pp.50-50
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• 2002

배추무사마귀병 저항성 유전 양식을 증명하기 위해서 CR계 F1에서 유래된 F2 세대를 포장시험과 유묘 검정을 실시하였다. F$_2$ 세대의 7 집단은 단인자우성으로 3:1의 분리비를 보였고, 5 집단은 중복 유전자가 관여하는 9:7의 유전 분리비를 보였다. 배추무사마귀병 저항성 유전자와 연관된 DNA 마커를 개발하기 위하여 CR-Saerona F$_2$ 집단을 배추무사마귀병 발병포장에서 재배하여 저항성 평가를 하였다. 220개의 임의의 프라이머를 이용하여 BSA-RAPD (Bulked segregant analysis-Randomly amplified polymorphic DNA)를 수행하였지만 CR-Saerona F2 집단에서 배추무사마귀병 저항성 유전자와 꼭 들어맞는 DNA 마커는 발견되지 않았다. 300개의 임의의 프라이머를 이용하여 CR-Saerona에서 유래된 F$_2$ 세대를 QTL 분석하였다. 저항성 정도는 발병지수에 따라 조사되었고 QTL 분석을 위해 one-way ANOVA 테스트를 하였다. 통계분석 결과 두 프라이머(K16-1, L2-2)가 저항성과의 상관관계를 보여 주었으나 유의성은 인정되지 않았다.

• Korean Journal of Plant Resources
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• v.16 no.3
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• pp.251-256
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• 2003

Genetic variation in field grown Korean ginseng(Panax ginseng C.A.Meyer) was evaluated using random amplified polymorphic(RAPD) markers. (This experiment was carried to collect the local native from farm of Chungnam National University in Korea in order to investigate genetic variation.) Some morphological characters showed considerable variation ranging 22 to 68cm in plant hight, 10 to 38mm in root diameter, 16 to 86g in root weight, and culum color and flowering date, respectively. Ten RAPD primers out of the 32 which produced reproducible bands in 662 Korean ginseng plants were selected for the further study. The total number of bands generated by 10 primers were 108 and among them 103 were polymorphic among the 662 plants with the polymorphism ratio of 94.5％. A total of 662 plants were classified into 16 groups based on polymorphic data with an URP 05 primer.

• 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.

• Horticultural Science & Technology
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• v.33 no.4
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• pp.598-604
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• 2015

This study was carried out to develop molecular techniques to allow the selection of zygotic seedlings in the early stage of the plant development. We identified 37 pollen-specific molecular markers from RAPD analysis and successfully used them for identification of the zygotic seedlings from various hybrid crosses. Three Satsuma mandarin cultivars ('Morita unshiu', 'Nangan 20' and 'Miyagawawase') were used as mother parents and seven cultivars ('Ponkan', 'Lee', 'Kinokuni', 'Shiranuhi', 'Tamnaneunbong', 'Shinyegam', and 'Sunburst' mandarins) served as pollen parents. PCR analysis showed that 2 primers could identify zygotic hybrid seedlings. Among them, an UBC-27 primer was used to identify the zygotic seedlings from hybrid crosses of "'Nangan 20' ${\times}$ 'Kinokuni'" mandarin, "'Nangan 20 ${\times}$ Ponkan'" mandarin and "'Miyagawawase ${\times}$ Sunburst'" tangerine. In total 29 out of 40 seedlings (73%), 9 out of 47 seedlings (19%), and 13 out of 45 (29%) were identified as zygotic seedlings, respectively. These results can show that the pollen-specific markers selected in this study can be used effectively for early identification of zygotic seedlings from Citrus hybrid crosses.

• 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.