• Title/Summary/Keyword: PCR-based DNA fingerprinting

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Intraspecific variations of the Yam (Dioscorea alata L.) based on external morphology and DNA marker analysis

  • Chang, Kwang-Jin;Yoo, Ki-Oug;Park, Cheol-Ho;Lim, Hak-Tae;Michio Onjo;Park, Byoung-Jae
    • Plant Resources
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    • v.3 no.3
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    • pp.211-218
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    • 2000
  • Intraspecific genetic relationship of 19 variation types of the Yam (Dioscorea alata) classified by their external morphological characteristics such as leaf and tuber shape were assessed by DNA using random and specific primer. 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 success 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. These DNA data were not matched well with those of morphological characters since they were divided into two major groups at the similarity coefficient value of 0.70. Therefore, Grouping of species into variation types by mainly morphological charactistics was suggested unreasonable.

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Genotypic and Phenotypic Diversity of PGPR Fluorescent Pseudomonads Isolated from the Rhizosphere of Sugarcane (Saccharum officinarum L.)

  • Rameshkumar, Neelamegam;Ayyadurai, Niraikulam;Kayalvizhi, Nagarajan;Gunasekaran, Paramsamy
    • Journal of Microbiology and Biotechnology
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    • v.22 no.1
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    • pp.13-24
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    • 2012
  • The genetic diversity of plant growth-promoting rhizobacterial (PGPR) fluorescent pseudomonads associated with the sugarcane (Saccharum officinarum L.) rhizosphere was analyzed. Selected isolates were screened for plant growthpromoting properties including production of indole acetic acid, phosphate solubilization, denitrification ability, and production of antifungal metabolites. Furthermore, 16S rDNA sequence analysis was performed to identify and differentiate these isolates. Based on 16S rDNA sequence similarity, the isolates were designated as Pseudomonas plecoglossicida, P. fluorescens, P. libaniensis, and P. aeruginosa. Differentiation of isolates belonging to the same group was achieved through different genomic DNA fingerprinting techniques, including randomly amplified polymorphic DNA (RAPD), amplified ribosomal DNA restriction analysis (ARDRA), repetitive extragenic palindromic (REP), enterobacterial repetitive intergenic consensus (ERIC), and bacterial repetitive BOX elements (BOX) analyses. The genetic diversity observed among the isolates and rep-PCR-generated fingerprinting patterns revealed that PGPR fluorescent pseudomonads are associated with the rhizosphere of sugarcane and that P. plecoglossicida is a dominant species. The knowledge obtained herein regarding the genetic and functional diversity of fluorescent pseudomonads associated with the sugarcane rhizosphere is useful for understanding their ecological role and potential utilization in sustainable agriculture.

DNA Fingerprinting Analysis of the Genus Phytophthora in Korea

  • Park, Dong-Suk;Kang, Hee-Wan;Lee, Mi-Hee;Park, Young-Jin;Lee, Byoung-Moo;Hahn, Jang-Ho;Go, Seung-Joo
    • Mycobiology
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    • v.31 no.4
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    • pp.235-247
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    • 2003
  • In order to investigate biodiversity and establish identification system for Phytophthora spp. in Korea, a variety of band pattern was produced by using the URP(universal rice primer). The fingerprint patterns of Phytophthora spp. showed many common and variable fragments according to their isolates in distinct genotypes. In particular, P. drechsleri was classified into four distinct types(I to IV). P. drechsleri(KACC 40498 and KACC 40499) and P. cryptogea(KACC 40413) appeared to have almost equal bands despite their being different species. Ninety isolates of Phytophthora spp. were clustered into 13 groups based on UPGMA(unweighted pair group method with arithmetic means) analysis. These DNA fingerprinting data would be helpful for inter- and intra-species identification of Phytophthora species.

Distribution and characteristics of Staphylococcus aureus subtypes isolated from dairy herds (젖소 목장에서 분리된 황색포도상구균의 아형 분포와 특성)

  • Yoo, Jong-hyun;Park, Hee-myung;Oh, Tae-ho;Sohn, Dae-ho;Han, Hong-ryul
    • Korean Journal of Veterinary Research
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    • v.39 no.5
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    • pp.995-1005
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    • 1999
  • Staphylococcus aureus is one of most prevalent intramammary pathogens and have characteristics which are not easily eradicated. Recently, to understand the sources and transmission of S aureus, many studies have focused on the subtyping of field isolate. This study was preformed to investigate the distribution pattern and characteristics of the isolates using phenotyping and genotyping. Samples were collected from milk of each udder, cow bodies (perianal region, vagina, tail, udder skin, sole) and environment (floor, liner, milker's hands, water, towel, insect) from 6 herds located in Kyung-gi province. Forty five strains of S aureus were isolated from 3 dairy herds (A, B, C) and were typed by hemolytic pattern, antibiotic resistant pattern, enterotoxin typing and PCR-based DNA fingerprinting. Slime productivity was also compared by each subtype to examine potential infectiousness. Of 45 strains, 41 were isolated from milk samples and 4 were isolated from liners. No strains isolated in the bodies and environment. Forty five strains isolated were classified as 18 subtypes by phenotyping and genotyping. There was common subtype between A and B herd, but the subtype of C herd showed different pattern. Among predominant subtypes, 60% of S aureus strain isolated from A and B herd showed subtype I and 50% of S aureus strain isolated from C herd belonged to subtype VI and X II. Neither somatic cell count (SCC) nor slime production was significantly different between predominant and minor subtypes. In summary, the study revealed that liners play more important roles in the mode of transmission than environmental sources. Several subtypes can be found in a herd, only a few subtype, however, was largely associated with the majority of infection.

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DNA Profiles of Trichoderma spp. in Korea

  • Park, Dong-Suk;Kang, Hee-Wan;Park, Young-Jin;Lee, Mi-Hee;Lee, Byoung-Moo;Hahn, Jang-Ho;Go, Seung-Joo
    • Mycobiology
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    • v.32 no.1
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    • pp.24-34
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    • 2004
  • Molecular approaches, internal transcribed spacer(ITS) sequences of ribosomal DNA, and Universal Rice Primer Polymerase Chain Reaction(URP-PCR) were used to investigate the genetic diversity, taxonomic complexity, and relationships of Trichoderma species in mushroom farms. Forty-one isolates of 13 Trichoderma spp. were used in this study and clustered into eight groups. The DNA fingerprint patterns and ITS1 region sequence alignment data showed similar results, but not in some species, such as T. virens, T. atroviride, T. harzianum, and T. aureoviride. Results of this study have proven that the morphology-based taxonomic system has some limitations in terms of classification. The data obtained in this study would be a good index for classifying indistinguishable Trichoderma strains.

Molecular Typing of Vibrio parahaemolyticus by Repetitive Element-PCR (rep-PCR) (Repetitive Element-PCR (rep-PCR)을 이용한 Vibrio parahaemolyticus 의 분자유전학적 아형 분류)

  • Kim, Won Sik;Hong, Seung Bok;Lee, Kyung;Lee, Jung Nam;Shin, Kyeong Seob
    • Korean Journal of Clinical Laboratory Science
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    • v.36 no.1
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    • pp.1-6
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    • 2004
  • The enterobacterial repetitive intergenic consensus (ERIC)-PCR is a recently described DNA fingerprinting technique based on amplification of repetitive element distributed in bacteria. We applied of ERIC-PCR to clinical isolates of Vibrio parahaemolyticus and other bacteria associated diarrhea. Twenty isolates of V. parahaemolyticus were used for intragenic genotyping, which were isolated from 2001 to 2002 in Chungbuk National University hospital. For interspecies genotyping, V. vulnificus, V. alginolyticus, V. parahaemolyticus, Escherichia coli, Salmonella and Shigella spp. were used. The genotyping were analyzed by ERIC-PCR. The genotyping of V. parahaemolyticus were grouped two major pattern (A, B) and were subdivided into 10 subtypes (A1, A2, B1-B8) by ERIC-PCR. These method distinctly differentiated bacterial species associated diarrhea. Those results show that ERIC-PCR can be reliable and efficient method for genotyping of V. parahaemolyticus and bacteria associated diarrhea.

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Comparative studies of the five edible mountain vegetables by DNA marker fingerprinting (DNA marker 지문법에 의한 취나물 5종 (청옥취 , 개미취 , 참취 , 수리취 , 곰취)의 비교연구)

  • 유기억
    • Korean Journal of Plant Resources
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    • v.9 no.3
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    • pp.305-310
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    • 1996
  • Five edible mountain vegetables(Saussurea sp. Aster tataricus, A. scaber. Synurus deltoides, Ligularia fischeri) were investigated on the basis of amplified DNA polymorphisms resulted from PCR (polymerase chain reaction) analysis. The sampled plants consisted of 38 individuals in 5 taxa. Only 10 primers out of 62 primers (60 random [10-mer] primers, two 15-mer [M13 core sequence, and (GGAT) sequence]) tested gave rise to polymorphisms in all of the tested plants, producing 176 DAN fragments amplified. Intraspecific polymorphisms found in each taxa showed intraspecies constancy (31.1-61.1%) in the banding patterns of individual plants: Saussurea sp. 31.1%, 15 bands, Aster tataricus, 40.9%, 18 bands, A. scaber. 38.5%, 15 bands. Synurus deltoides, 34.7%, 17 bands, and Ligularia fischeri, 61.1%, 22 brands, respectively. All five species were well classified from each other at the 0.93 level of similarity index value. Intraspecific and interspecific variations were appeared at the levels ranging from 0.62 to 0.99. Based on these results, our PCR analyses support the previous data derived from external morphology of the 5 edible mountain vegetables, but very low levels o intraspecific variations were detected in all of these taxa.

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PCR Approach for Detection and Typing of Epidermodysplasia Verruciformis-associated Human Papillomavirus Types

  • Shin, Yong-Jae;Choi, Hyun-Chul;Kim, Chan-Gil;Lee, Kwang-Ho
    • The Journal of Korean Society of Virology
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    • v.29 no.1
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    • pp.39-44
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    • 1999
  • Warts, or verrucae, are benign epithelial proliferations of the skin and mucosa caused by infection with human papillomaviruses (HPV). It is now recognized that there are many different HPV types. Especially type3 is most frequently observed in flat wart. Other types, such as type2, 10, 14, 27, 28, 29, 38, and 41 are rarely encounted in flat wart. We describe here a simple and economic method for detection and identification of epidermodysplasia verruciformis-associated HPV. The method is based on polymerase chain reaction (PCR) amplification and restriction analysis. The method has been developed with cloned HPV DNA and DNA from clinical samples. Clinical samples are from either frozen tissue or paraffin-embedded tissue. Genomic fragments were obtained from two different HPV types (3 and 10). The amplification fragments were identified by a form of miniature fingerprinting, with a set of restriction enzymes that gave a unique digestion pattern for each HPV type. We have tested 74 clinical samples. Only type3 among these clinical samples is detected, and one sample is involved in neither type3 nor type10.

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Two Genetically Distinct Groups of Acidovorax citrulli are Present in Watermelon-growing Fields in Korea

  • Choi, Okhee;Cho, Su Kyung;Kang, Byeongsam;Cho, Jaeyeong;Park, Jiyeong;Lee, Yeyeong;Kim, Jinwoo
    • Journal of agriculture & life science
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    • v.50 no.2
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    • pp.53-59
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    • 2016
  • Bacterial fruit blotch(BFB) of cucurbits caused by Acidovorax citrulli(Acc) continues to diminish fruit yields. The aim of this study was to address whether two genetically distinct populations of Acc are present in watermelon-growing fields in Korea. For this purpose, we used the enterobacterial repetitive intergenic consensus polymerase chain reaction(ERIC-PCR) profiling and substrate-utilization profiles. According to the results of ERIC-PCR, group I and II strains showed clearly differentiated PCR-based fingerprinting profiles. Differences between group I and II strains included amplification of unique, group-specific DNA fragments such as the 1.3-, 0.28-, and 0.25-kb fragments in ERIC-PCR. Acc stains belonging to group I did not use L-leucine, whereas group II strains did use the substrate. Our results support the genetic differentiation of Acc strains into two groups and demonstrate that Acc strains from both groups are previously existed in watermelon-growing fields in Korea. Information about the genetic diversity of Acc under the present study will help scientists and managers form strategies to control BFB.

Differentiation Among Commercial Strains of Pleurotus spp. Based on DNA Fingerprinting Using Universal Rice Primer (URP) (Universal Rice Primer(URP)에 의한 DNA 핵산지문법을 이용한 느타리의 유통 품종간 구분)

  • Seo, Kyoung-In;Jang, Kab-Yeul;Yoo, Young-Bok;Park, Soon-Young;Kim, Kwang-Ho;Kong, Won-Sik
    • The Korean Journal of Mycology
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    • v.36 no.2
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    • pp.130-137
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    • 2008
  • To distinct the commercial strains in Pleurotus spp., 81 strains in eight Pleurotus species were used. DNA fingerprinting using URP-PCR was conducted to determine the phylogenetic relationships among Pleurotus strains. DNA profiles of Pleurotus species obtained by twelve URP primers were analyzed for genetic similarity by NTSYS program. We could divide strains into ten clusters, in which three of them belong to P. ostreatus and the others to the different species, respectively. At the 76% similarity level, 70 P. ostreatus strains were distinguished into three clusters. Cluster I contained 35 strains and some of them showed almost 100% similarity, one strain closely related to Weonhyeong and six strains closely related to Wangheukpyeong. In cluster II, twenty-one out of 23 strains showed 100% to Suhan. Cluster III contained twelve strains, including six strains closely related to Chunchu-2. The results suggested that there are many same strains with different names in mushroom spawn market.