• Title/Summary/Keyword: Random Amplification of Polymorphic DNA

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Development of a SCAR Marker for Sex Identification in Asparagus

  • Kim, Seong-Cheol;Jung, Yong-Hwan;Seong, Ki-Cheol;Chun, Seung-Jong;Kim, Chun Hwan;Lim, Chan Kyu;Joa, Jae-Ho;Lee, Dong-Sun
    • Korean Journal of Plant Resources
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    • v.27 no.3
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    • pp.236-241
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    • 2014
  • A sex-linked random amplified polymorphic DNA (RAPD) marker was identified from Asparagus officinalis L. and was converted into a sequence-characterized amplified regions (SCAR) marker for the large-scale screening of male and female plants. A total of 100 arbitrary decamer oligonucleotide primers were used for the RAPD analysis. Among them, the primer UBC347 amplified one female-specific 400 base pair DNA. Subsequently, the amplified RAPD fragment was cloned and sequenced. The fragment was abundant in AT and shared sequence homology with retrotransposon elements. On the basis of the sequence obtained, a pair of SCAR primer was designed. The amplification product, named F400, was the same size as the respective RAPD fragment from which it was derived. The F400 SCAR marker resulted to be female-specific in the three asparagus varieties tested in this study. This SCAR marker can be used for an early and rapid identification of female and male plants during breeding programs of asparagus.

Development of a sequence-characterized amplified region (SCAR) marker for female off-season flowering detection in date palm (Phoenix dactylifera L.)

  • Lalita Kethirun;Puangpaka Umpunjun;Ngarmnij Chuenboonngarm;Unchera Viboonjun
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.190-199
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    • 2023
  • Date palm (Phoenix dactylifera L.: Arecaceae) is a dioecious species where only female trees bear fruits. In their natural state, date palms produce dates once a year. However, in Thailand, some trees were observed to produce dates during the off-season, despite no variations in morphology. The availability of such off-season fruits can significantly increase their market value. Interestingly, most female off-season date palms investigated in this study were obtained through micropropagation. Hence, there is an urgent need for genetic markers to distinguish female offseason flowering plantlets within tissue culture systems. In this study, we aimed to develop random amplification of polymorphic DNA-sequence characterized amplified region (RAPD-SCAR) markers for the identification of female off-season flowering date palms cultivated in Thailand. A total of 160 random decamer primers were employed to screen for specific RAPD markers in off-season flowering male and female populations. Out of these, only one primer, OPN-02, generated distinct genomic DNA patterns in female off-season flowering (FOFdp) individuals compared to female seasonal flowering genotypes. Based on the RAPD-specific sequence, specific SCAR primers denoted as FOFdpF and FOFdpR were developed. These SCAR primers amplified a single 517-bp DNA fragment, predominantly found in off-season flowering populations, with an accuracy rate of 60%. These findings underscore the potential of SCAR marker technology for tracking offseason flowering in date palms. Notably, a BLAST analysis revealed a substantial similarity between the SCAR marker sequence and the transcript variant mRNA from Phoenix dactylifera encoding the SET DOMAIN GROUP 40 protein. In Arabidopsis, this protein is involved in the epigenetic regulation of flowering time. The genetic potential of the off-season flowering traits warrants further elucidation.

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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Characteristics of Dioscorea alata L. Introduced from Tropical and Subtropical Regions (도입 마(Dioscorea alata L.)의 특성 분석)

  • Chang, K.J.;Yoo, K.O.;Park, C.H.;Park, J.I.;Hong, K.H.;Park, J.H.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.3 no.1
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    • pp.48-69
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    • 2001
  • A lot of clones of the genus Dioscorea have been introduced from some tropical and subtropical regions since 1997. In 33 clones of water yams (Dioscorea alata L.), some morphological characteristics were investigated at the field. Variation ranges of the total weight and tuber number per stump were within the ranges from 90 to 2,147 g with an average of 610 g ; and 1.3-4.7 with an average of 2.8, respectively. The color tones observed on the tuber-flesh were sorted into 3 color-categories, i.e., white, pale brown and pale purple, and those on leaves were sorted into 3 color-categories, i.e., green, heavy green and purplish green. Intraspecific genetic relationship of 19 variation types of the Yam classified by their external morphological characteristics such as leaf and tuber shape was assessed by DNA using random and specific primers. Twenty two out of 113 primers (100 random[10-mer] primers, two 15 mer [M13 core sequence, and (GGAT)4 sequence]) had been used in PCR-amplification. Only 12 primers, however, were successful in DNA amplification in all of the analyzed plants, resulting in 93 randomly and specifically amplified DNA fragments. The analyzed taxa showed very high polymorphisms(69 bands, 71.0%), allowing individual taxon to be identified based on DNA fingerprinting. Monomorphic bands among total amplified DNA bands of each primer was low under the 50%. Similarity indices between accessions were computed from PCR(polymerase chain reaction) data, and genetic relationships among intraspecific variations were closely related at the levels ranging from 0.66 to 0.90.

Genetic diversity analysis of high yielding rice (Oryza sativa) varieties cultivated in Bangladesh

  • Epe, Isma Akter;Bir, Md. Shahidul Haque;Choudhury, Abul Kashem;Khatun, Asma;Aktar, Most Mohshina;Arefin, Md. Shamsul;Islam, Mohammed Aminul;Park, Kee Woong
    • Korean Journal of Agricultural Science
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    • v.48 no.2
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    • pp.283-297
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    • 2021
  • Investigation of genetic diversity and molecular characterization in high yielding rice varieties is important for their identification. The experiment was conducted during 2016 - 2017 to analyse the genetic diversity of fifteen high yielding rice varieties in Bangladesh by using random amplification of polymorphic DNA (RAPD) markers. Polymorphism was revealed in 12 RAPD primers out of 30, whereas no other reaction was detected on the remaining 18 primers. The 40 out of 45 bands (88.89%) polymorphics were produced by the primers and ranged from 50 to 100%. The maximum number of polymorphic bands was produced by the primer OPB-18 whereas the lowest number of polymorphic bands belonged to OPC-12. The genetic similarity coefficients were determined with the RAPD data, which ranged from 0.47 to 0.94. The unweighted paired group of arithmetic means (UPGMA) dendrogram presented the studied rice varieties into two major clusters. Moreover, the value of Nei's genetic diversity is 0.26 and the Shanon information index is 0.41. The study produced distinct positions, suggesting that the genotypes were different from each other. The results indicated that these markers could be efficient for comparing the genetic relationships, patterns of variation, and measurement of genetic distance among rice varieties. Considering all of these results, RAPD analysis is found to be an effective tool for estimating the genetic diversity of different rice varieties. The outcomes of this research may contribute to the germplasm data of rice accessions and a future breeding program of rice genotypes.

Randomly Amplified Polymorphic DNA Analyses of Pestalotiopsis theae Isolated from Sweet Persimon (재배되는 단감나무로 부터 분리한 Pestalotiopsis theae의 RAPD 기법을 이용한 유전특성의 비교분석)

  • Lee, Youn-Su;Woo, Su-Jin;Choi, Hei-Sun;Kim, Kyoung-Su;Kang, Won-Hee;Kim, Myoung-Jo;Shim, Jae-Ouk;Chang, Tae-Hyun;Lim, Tae-Heon
    • The Korean Journal of Mycology
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    • v.26 no.3 s.86
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    • pp.365-372
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    • 1998
  • In this study, we evaluated the genetic relationships of fourty seven Pestalotiopsis theae isolates collected from diseased sweet persimon in various places in southern part of Korea using RAPD (Randomly Amplified Polymorphic DNAs) method. As a result of the amplification, eight primers showed total of 86 bands ranging from 0.3 Kb to 3.2 Kb. Among those 86 bands, 84 polymorphic bands were used for bionominal matrix code (0, 1), and UPGMA dendrogram analysis. Similarities among the compared isolates ranged from below 60% to more than 95%. Most of the compared isolates showed $50{\sim}80%$ similarities. The number of isolate pairs which showed more than 80% similarity were 248. The number of isolate pairs which showed $50{\sim}80%$ similarity were 789, and the number of isolate pairs which showed below 50% similarity were 21. Isolate SP-21 (No.9) showed below 50% similarity with all the isolates compared. At 50% similarity level, all the isolates compared, except isolate SP-21 (No.9), were included in one big group. At 65% similarity level, all the isolates compared, except isolate SP-21 (No.9), were divided into three different groups. At 75% similarity level, all the isolates compared, except isolates SP-47 (No. 23) and SP-21 (No.9), were divided into six different groups.

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Analysis of Genetic Relatedness by Random Amplified Polymorphic DNA (RAPD) in Pecan Taxa (RAPD를 이용한 Pecan 품종의 유전적 관계 분석)

  • 신동영;김회택;박종인;노일섭
    • Korean Journal of Plant Resources
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    • v.13 no.1
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    • pp.1-10
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    • 2000
  • Pecan is deciduous tree and belongs to the Julandaceae family. Pecan is an economically important as a nut and timber crop. Heterozygosity is expected to be high for typically cross-pollinated. Yet little is known about the nature of genetic variation within this species. In addition, the pedigree of many pecan cultivars remains unknown or is questionable. In this study, the phylogenetic relationships between 22 pecan cultivars and its analyzed by RAPD (randomly amplified polymorphic DNA). PCR Amplification used 40 randomly selected oligoes as primers. Based on their genetic similarities derived from the RAPD data, the 22 pecan cultivars were classified into different five groups in agarose gel. The 22 pecan cultivars were classified into five sectional groups by UPGMA clustering analysis, too. C. flacra and Black walnut showed the 0.9 of similarity index and Farley, Pawnee showed the 0.85 of similarity index. The 22 pecan cultivars were classified into different five groups by analysis of the 4% polyacrylamide gel fraction. (Group I : 1, 2, 3, 4, 13, 16, 17, 20, 21 Group II : 14,18 GroupIII : 6,12 GroupIV : 5, 11, 15, 19, 22 CroupV : 7, 8, 9, 10) Group V show the 1.0 of similarity index and Farley, Sturya, Clarke, Pawnee show the 0.98 of similarity index and Kiowa, Schley show the 0.92 of similarity index. Results from this study indicated that RAPD can be used to establish the genetic relationships among the 22 pecan cultivars. Similarity coefficients generally agreed with what would be predicted in cultivars with known pedigrees, and we could accurately construct relationships among cultivars. In addition, we have shown that RAPD provides useful information on the origin of unknown cultivars.

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Breeding and characterization of a long shelf-life cultivar 'DanBi 7Ho' by mono-mono crossing in Pleurotus eryngii

  • Kim, Min Keun;Sim, Soon Ae;Park, Ji Hye;Ryu, Jae San;Choi, Si Lim;Hong, Kwang Pyo
    • Journal of Mushroom
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    • v.16 no.3
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    • pp.140-146
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    • 2018
  • Pleurotus eryngii is one of the most commercially important mushrooms cultivated in Korea. However, the shelf-life of the fruiting body is short, limiting its export. A new hybrid strain H17 of P. eryngii was developed to extend the shelf-life by mono-mono crossing between monokaryotic strains derived from DanBi and KNR2774. Although the cultivation period of H17 was slightly longer than that of the reference cultivar Kenneutari No.2, the quality did not change and remained normal after a period of 65.0 days at $4^{\circ}C$. This result was significantly different from that of the reference cultivar Kenneutari No.2. Analysis of the genetic characteristics of the new hybrid strain H17 revealed a different profile from that of the parental and reference cultivars when random amplification of polymorphic DNA (RAPD) primers was used. These results demonstrate that H17 is a new cultivar with improved storability after harvesting.

Characterization of Quinolone-Resistant Clinical Isolates of Escherichia coil in Korea

  • Lee, Yeonhee;Yoojung Oh;Park, Seohyung;Misun Ha
    • Journal of Microbiology
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    • v.40 no.2
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    • pp.98-103
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    • 2002
  • Twenty-eight clinical isolates of Escherichia coil, composed of thirteen norfloxacin resistant isolates (MIC of >16${\mu}$g/ml), one intermediately resistant isolate (MIC of 8${\mu}$g/ml), and fourteen susceptible isolates (MIC of <4${\mu}$g/ml), were randomly selected to study the norfloxacin resistance mechanism and phylogeny in clinical isolates in Korea. Eleven nofloxacin resistant isolates and one susceptible isolate were multi-drug resistant (MDR). Every norfloxacin resistant isolate with MIC higher than 32${\mu}$g/ml had the same three mutations: Ser83\longrightarrowLeu and Asp87\longrightarrowAsn or Tyr in GyrA and Ser80\longrightarrowIle in ParC. Whereas a resistant isolate with MIC of 16${\mu}$g/ml had three mutations but Asp87 in GyrA was replaced with Gly instead of Asn. The intermediately resistant isolate had the same two mutations in GyrA but a different mutation in ParC, Glu84\longrightarrowLys. Among the susceptible isolates, two isolates with MIC of 4${\mu}$g/ml had one mutation: Ser83\longrightarrowiLeu in GyrA, and no mutation was found in the susceptible isolates. Resistant isolates showed higher efflux activity than the susceptible ones, with random amplification of polymorphic DNA (RAPD), six susceptible isolates form a separate group from the rest of the isolates.

Intraspecific genetic variation in Corynandra chelidonii (Angiosperms: Cleomaceae) as revealed by SCoT, ISSR and RAPD analyses

  • Sirangi, Subash;Jogam, Phanikanth;Nemali, Gandhi;Ajmeera, Ragan;Abbagani, Sadanandam;Raju, Vatsavaya S.
    • Journal of Plant Biotechnology
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    • v.47 no.4
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    • pp.289-297
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    • 2020
  • The genetic diversity of two subpopulations of Corynandra chelidonii, one of terrestrial and the other of aquatic environments, was measured with molecular markers, such as start codon targeted (SCoT), inter simple sequence repeats (ISSR), and random amplification of polymorphic DNA (RAPD). The traditional morphological traits such as habitat, habit, leaf morphology, the colour of the sepals and petals, number of stamens, and seed morphology formed the base for their realization as two varieties, C. chelidonii var. pallae and C. chelidonii var. chelidonii. The polymorphism between the two variants was 100% with the primers SCoT-2 and OPA-1 and 4, while maximum polymorphism was detected with ISSR-2, SCoT-3, and OPA-3. The study used, for the first time, more than one molecular marker to assess the genetic variation underscoring the morphological variation in Corynandra chelidonii (L.f.) Cochrane & Iltis. The study justifies the recognition of the two subpopulations of Corynandra chelidonii from aquatic and terrestrial environments as two distinct varieties, C. chelidonii var. pallae (Reddy & Raju) V.S.Raju and C. chelidonii var. chelidonii, respectively, based on the traditional taxonomic evidence.