• Title/Summary/Keyword: Polymorphic markers

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AFLP analysis to assess genomic stability in Solanum regenerants derived from wild and cultivated species

  • Aversano, Riccardo;Di Dato, Francesco;Di Matteo, Antonio;Frusciante, Luigi;Carputo, Domenico
    • Plant Biotechnology Reports
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    • v.5 no.3
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    • pp.265-271
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    • 2011
  • The cultivated potato as well as its tuber-bearing relatives are considered model plants for cell and tissue culture, and therefore for exploiting the genetic variation induced by in vitro culture. The association between molecular stability and tissue culture in different genetic backgrounds and ploidy levels has already been explored. However, it still remains to be ascertained whether somaclonal variation differs between callus-derived chromosome-doubled and undoubled regenerants. Our research aimed at investigating, through amplified fragment length polymorphism (AFLP) markers, the genetic changes in marker-banding patterns of diploid and tetraploid regenerants obtained from one clone each of Solanum bulbocastanum Dunal and S. cardiophyllum Lindl (both 2n = 2x = 24) and tetraploids from cultivated S. tuberosum L. (2n = 4x = 48). Pairwise comparisons between the banding patterns of regenerants and parents allowed detecting considerable changes associated to in vitro culture both at diploid and tetraploid level. The percentages of polymorphic bands between diploid and tetraploid regenerants were, respectively, 57 and 69% in S. bulbocastanum and 58 and 63% in S. cardiophyllum. On average, the frequencies of lost parental fragments in regenerants were significantly higher than novel bands both in S. bulbocastanum (48 vs. 22%) and S. tuberosum (36 vs. 18%) regenerants. By contrast, in S. cardiophyllum, a similar incidence of the two events was detected (32 vs. 29%). Our results revealed that structural changes after tissue culture process strongly affected the genome of the species studied, but diploid and tetraploids regenerated plants responded equally.

Population analysis of eelgrass, Zostera marina L. in Geojedo, Gaedo, and Jedo on the southern coastal water of Korea using RAPD-PCR (RAPD 방법을 이용한 거제도, 개도, 제도해역에서 채집한 말잘피 개체분석)

  • Cho, Eun-Seob;Lee, Sang-Yong;Kim, Jeong-Bae
    • Journal of Life Science
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    • v.17 no.4 s.84
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    • pp.455-461
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    • 2007
  • Assessments of population genetic structure and diversity can be of value in formulating management plans for threatened eelgrass(Zostera maim). Using randomly amplified polymorphic DNA markers, we found evidence of significant genetic structure among the populations of eelgrass sampled at three areas(Geojedo, Gaedo, and Jedo). A highly isolated(>100 km apart) population from the Geojedo had a long genetic distance(0.16), whereas the populations from the Gaedo and Jedo(<10 km apart) exhibited far less distance(0.08). The analysis of similarity within population showed that Geojedo was over 70%, which was of lower value than of Gaedo and Jedo. Based on these results, we realized that heterogeneous population was in accordance with geographic separation. This is caused by limited seed dispersal and interrupted gene flow, although the sample size is small.

Limits of Direct PCR Amplification from Seaweeds Using Arbitrary and ITS Primers (해조류로부터 Arbitrary 및 ITS Primer들을 사용한 직접 PCR 유전자 증폭반응의 한계)

  • 김용국;진형주;박선미;진덕희;홍용기
    • Journal of Life Science
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    • v.9 no.1
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    • pp.15-21
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    • 1999
  • The random amplified ploymorphic DNAs (RAPD) assay is a simple and useful tool in identification of appropriate genetic markers, that requires no knowledge of target DNA sequence. RAPD products were generated directly from seaweed tissues, without prior nucleic acid extraction, of Porphyra yezoensis, Ulva pertusa and Undaria pinnatifida. The nuclear rDNA internal transcribed spacer (ITS) fragment however was not amplified directly from the seaweed tissues. Using DNA extracted by the LiCl method, both the ITS and RAPD's have been amplified by the polymerase chain reaction. RAPD of P yezoensis, thallus (n) and conchocelis (2n) produced lots of different polymorphic bands (36-50$\%$) depending on the arbitrary primers used. Difference was also observed between direct tissues amplification and DNA extracts amplification (53-57$\%$). Thus it is important to use the same ploidy of tissue for DNA extraction and as a RAPD template.

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Characterization of Novel Trichoderma asperellum Isolates to Select Effective Biocontrol Agents Against Tomato Fusarium Wilt

  • El_Komy, Mahmoud H.;Saleh, Amgad A.;Eranthodi, Anas;Molan, Younes Y.
    • The Plant Pathology Journal
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    • v.31 no.1
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    • pp.50-60
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    • 2015
  • The use of novel isolates of Trichoderma with efficient antagonistic capacity against Fusarium oxysporum f. sp. lycopersici (FOL) is a promising alternative strategy to pesticides for tomato wilt management. We evaluated the antagonistic activity of 30 isolates of T. asperellum against 4 different isolates of FOL. The production of extracellular cell wall degrading enzymes of the antagonistic isolates was also measured. The random amplified polymorphic DNA (RAPD) method was applied to assess the genetic variability among the T. asperellum isolates. All of the T. asperellum isolates significantly reduced the mycelial growth of FOL isolates but the amount of growth reduction varied significantly as well. There was a correlation between the antagonistic capacity of T. asperellum isolates towards FOL and their lytic enzyme production. Isolates showing high levels of chitinase and ${\beta}$-1,3-glucanase activities strongly inhibited the growth of FOL isolates. RAPD analysis showed a high level of genetic variation among T. asperellum isolates. The UPGMA dendrogram revealed that T. asperellum isolates could not be grouped by their antagonistic behavior or lytic enzymes production. Six isolates of T. asperellum were highly antagonistic towards FOL and potentially could be used in commercial agriculture to control tomato wilt. Our results are consistent with the conclusion that understanding the genetic variation within Trichoderma isolates and their biochemical capabilities are required for the selection of effective indigenous fungal strains for the use as biocontrol agents.

Genetic Diversity and Phylogenetic Relationship of Genus Phyllostachys by RAPD Markers (RAPD분자 마커를 이용한 왕대속 대나무의 유전적 다양성 및 계통 관계)

  • Lee, Song-Jin;Huh, Man-Kyu;Shin, Hyun-Cheol;Huh, Hong-Wook
    • Journal of Life Science
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    • v.20 no.6
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    • pp.819-824
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    • 2010
  • Genus Phyllostachys is a long-lived woody species primarily distributed throughout South East Asia. Many species of this genus has been regarded as medically and ecologically important in the world. We evaluated representative samples of the four taxa with RAPD to estimate genetic relationships within the genus Phyllostachys. The percentages of polymorphic loci were 8.9-33.3% at the species level. P. bambusoides was found to show lower genetic diversity (H=0.018) than other species. Total genetic diversity ($H_T$) was 0.315, genetic diversity within populations ($H_S$) was 0.043, the proportion of total genetic diversity partitioned among populations ($G_{ST}$) was 0.659 and the gene flow (Nm) was 0.0263. As some Korean populations were isolated and patchily distributed, they exhibited low levels of genetic diversity. The four taxa of the genus Phyllostachys analyzed were distinctly related to a monophyletic. P. nigra var. henonis. Stapf was found to be more closely related to P. pubescens than to P. nigra. P. bambusoides was quite distinct from the remaining species.

Development and characterization of microsatellite markers for an endangered species, $Epinephelus$ $bruneus$, to establish a conservation program

  • An, Hye-Suck;Kim, Jae-Woo;Lee, Jang-Wook;Kim, Shin-Kwon;Lee, Bae-Ik;Kim, Dae-Jung;Kim, Yi-Cheong
    • Animal cells and systems
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    • v.16 no.1
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    • pp.50-56
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    • 2012
  • Kelp grouper ($Epinephelus$ $bruneus$ Bloch 1793) is a commercially important fish in Korea. In recent years, the catch of kelp grouper in the coastal waters of Korea has significantly declined. Despite its importance, little is known about its genetic diversity and conservation efforts are hampered. In this study, we isolated and characterized 12 microsatellite loci using an enrichment method based on magnetic/biotin capture of microsatellite sequences from a size-selected genomic library. All loci were readily amplified and contained TG/CA denucleotide repeats. To characterize each locus, 30 individuals from a natural E. bruneus population in the coastal waters of Jeju Island, Korea, were genotyped. All loci except three, KEm118, KEm154, and KEm219, were polymorphic, with an average of 8.1 alleles per locus (range 2-18). The mean observed and expected heterozygosities were 0.47 (range 0.19-1.00) and 0.61 (range 0.29-0.92), respectively. A significant deviation from Hardy-Weinberg equilibrium was observed at three loci (KEm134, KEm184, and KEm283). These findings will be useful for effective monitoring and management of genetic variation of kelp grouper as well as for the implementation of a fisheries conservation program.

Genetic Variation in the Natural Populations of Abies holophylla Max. Based on RAPD Analysis (RAPD 분석(分析)에 의한 전나무 천연집단(天然集團)의 유전변이(遺傳變異))

  • Kim, In Sik;Hyun, Jung Oh
    • Journal of Korean Society of Forest Science
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    • v.88 no.3
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    • pp.408-418
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    • 1999
  • On the basin of RAPD analysis, genetic diversity and structure of the natural populations of Abies holophylla was estimated by AMOVA procedure. The average value of percent of polymorphic markers was 71.9%. Most variation existed among individuals within population(80.2%). Genetic differentiation among populations(${\Phi}_{ST}$) was 0.198. When the populations were grouped as two region(i.e., Taebaek and Sobaek Mountain Regions), 8.5% of the total genetic variation was explained as regional differences. The heterogeneity of molecular variance among populations was investigated with Bartlett's test, which revealed that populations of Mt. Taebaek and Mt. Gariwang were more heterogeneous. Generally, the populations of Taebaek Mountain Reion were more heterogeneous than those of Sobaek Mountain Reion. Finally, the applicability of AMOVA to the populations frenetic study was discussed in comparison with other measures of genetic differentiation which were widely used.

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Morphological Characteristics of Bemisia tabaci(Gennadius) (Homoptera: Aleyrodidae) and Discrimination of Their Biotypes in Korea by DNA Makers (담배가루이 Bemisia tabaci(Gennadius)(Homoptera: Aleyrodidae)의 형태적 특징과 DNA 표식자에 의한 biotype 판별)

  • 이명렬;안성복;조왕수
    • Korean journal of applied entomology
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    • v.39 no.1
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    • pp.5-12
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    • 2000
  • The sweetpotato whiteflies, Bemisia tabaci(Gennadius), were found recently in Korea on Glycine max, Euphorbia pulcherrima, and Rosa hybrida. The biotype identity of Bemisia tabaci in Korea was determined by several DNA markers including the random amplified polymorphic DNAs, and restriction fragments length polymorphism of mitochondrial 12S and 16S rRNA genes. The electromorph profiles of DNA fragments from the rose(Jincheon) and poinsettia(Seoul) populations in Korea are both identical to those of B biotypes distributed in Australia, Israel, and Japan. The populations of B. tabaci collected on Glycine max, Ipomea batatas, and Perilla frutescens in different localities retained the same DNA markes with the population from Lonicera japonica and shikoku of Japan. These populations are non-B biotype and considered as an indigenous type in the Far Eastern Asia Region including Korea and Japan, Morphological Characteristics of B. Tabaci were also observed by the scanning electron microscope and described with the comparison to the other important whitefly pest, Trialeurodes vaporariorum (Westwood).

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Preimplantation genetic diagnosis for Charcot-Marie-Tooth disease

  • Lee, Hyoung-Song;Kim, Min Jee;Ko, Duck Sung;Jeon, Eun Jin;Kim, Jin Young;Kang, Inn Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.40 no.4
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    • pp.163-168
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    • 2013
  • Objective: Preimplantation genetic diagnosis (PGD) is an assisted reproductive technique for couples carrying genetic risks. Charcot-Marie-Tooth (CMT) disease is the most common hereditary neuropathy, with a prevalence rate of 1/2,500. In this study, we report on our experience with PGD cycles performed for CMT types 1A and 2F. Methods: Before clinical PGD, we assessed the amplification rate and allele drop-out (ADO) rate of multiplex fluorescent polymerase chain reaction (PCR) followed by fragment analysis or sequencing using single lymphocytes. We performed six cycles of PGD for CMT1A and one cycle for CMT2F. Results: Two duplex and two triplex protocols were developed according to the available markers for each CMT1A couple. Depending on the PCR protocols, the amplification rates and ADO rates ranged from 90.0% to 98.3% and 0.0% to 11.1%, respectively. For CMT2F, the amplification rates and ADO rates were 93.3% and 4.8%, respectively. In case of CMT1A, 60 out of 63 embryos (95.2%) were diagnosed and 13 out of 21 unaffected embryos were transferred in five cycles. Two pregnancies were achieved and three babies were delivered without any complications. In the case of CMT2F, a total of eight embryos were analyzed and diagnosed. Seven embryos were diagnosed as unaffected and four embryos were transferred, resulting in a twin pregnancy. Two healthy babies were delivered. Conclusion: This is the first report of successful pregnancy and delivery after specific PGD for CMT disease in Korea. Our PGD procedure could provide healthy babies to couples with a high risk of transmitting genetic diseases.

Retrotransposon Microsatellite Amplified Polymorphism Strain Fingerprinting Markers Applicable to Various Mushroom Species

  • Le, Quy Vang;Won, Hyo-Kyung;Lee, Tae-Soo;Lee, Chang-Yun;Lee, Hyun-Sook;Ro, Hyeon-Su
    • Mycobiology
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    • v.36 no.3
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    • pp.161-166
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    • 2008
  • The retrotransposon marY1 is a gypsy family retroelement, which is detected ubiquitously within the fungal taxonomic groups in which mushrooms are included. To utilize marY1 as a molecular marker for the DNA fingerprinting of mushrooms, oligonucleotides marY1-LTR-L and marY1-LTR-R were designed on the basis of highly conserved regions from the multiple sequence alignment of 30 marY1 sequences retrieved from a nucleotide sequence database. In accordance with $\underline{Re}trotransposon$ $\underline{M}icrosatellite$ $\underline{A}mplified$ $\underline{P}olymorphism$ (REMAP) fingerprinting methodology, the two oligonucleotides were utilized together with the short sequence repeat primers UBC807 and UBC818 for polymerase chain reaction using templates from different mushroom genomic DNAs. Among the tested oligonucleotides, the marY1-LTR-L and UBC807 primer set yielded the greatest amount of abundance and variation in terms of DNA band numbers and patterns. This method was successfully applied to 10 mushroom species, and the primer set successfully discriminated between different commercial mushroom cultivars of the same strains of 14 Pleurotus ostreatus and 16 P. eryngii. REMAP reproducibility was superior to other popular DNA fingerprinting methodologies including the random amplified polymorphic DNA method.