• Title/Summary/Keyword: RAPD PCR

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Studies on the Genetic Diversity using RAPD in Leuconostoc sp. Isolated from Kimchi (김치에서 분리한 Leuconostoc속의 RAPD 법을 이용한 유전적 변이도 추론)

  • Kim, Eun-Kyung;Ryu, Chun-Sun;So, Myung-Hwan;Kim, Young-Bae
    • Korean Journal of Food Science and Technology
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    • v.29 no.5
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    • pp.1063-1066
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    • 1997
  • Thirty four Leuconostoc strains originated from kimchi were subjected to analyse their genetical similarity by using RAPD-PCR. At the 0.5 level of similarity, L. mesenteroides subsp. mesenteroides and L. mesenteroides subsp. dextranicum. formed 3 and 2 clusters, and each L. lactis and L. paramesenteroides strain was clustered in one, respectively. Every type strain tested as a control, belongs to the cluster or one of the clusters of the corresponding species or subspecies as the isolates from kimchi, respectively. However the similarity between a and b cluster was less than 0.2 and c cluster is even less similar to both of them (a and b). L. mesenteroides subsp. mesenteroides strains from kimchi have shown a varaiety of RAPD patterns in spite of morphological and physiological similarities.

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Genetic Diversity and Phylogenetic Relationships among Microsporidian Isolates from the Indian Tasar Silkworm, Antheraea mylitta, as Revealed by RAPD Fingerprinting Technique

  • Hassan, Wazid;Nath, B. Surendra
    • International Journal of Industrial Entomology and Biomaterials
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    • v.29 no.2
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    • pp.169-178
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    • 2014
  • In this study, we investigated genetic diversity of 22 microsporidian isolates infecting tropical tasar silkworm, Antheraea mylitta collected from various geographical forest locations in the state of Jharkhand, India, using polymerase chain reaction (PCR)-based marker assay: random amplified polymorphic DNA (RAPD). A type species, NIK-1s_mys was used as control for comparison. The shape of mature microsporidians was found to be oval to elongate, measuring 3.80 to $5.10{\mu}m$ in length and 2.56 to $3.30{\mu}m$ in width. Of the 20 RAPD primers screened, 16 primers generated reproducible profiles with 298 polymorphic fragments displaying high degree of polymorphism (97%). A total of 14 RAPD primers produced 45 unique putative genetic markers, which were used to differentiate the microsporidians. Calculation of genetic distance coefficients based on dice coefficient method and clustering with un-weighted pair group method using arithmetic average (UPGMA) analysis was conducted to unravel the genetic diversity of microsporidians infecting tasar silkworm. The similarity coefficients varied from 0.059 to 0.980. UPGMA analysis generated a dendrogram with four microsporidian groups, which appear to be different from each other as well as from NIK-1s_mys. Two-dimensional distribution based on Euclidean distance matrix also revealed considerable variability among different microsporidians identified from the tasar silkworms. Clustering of few microsporidian isolates was in accordance with the geographic origin. The results indicate that the RAPD profiles and specific/unique genetic markers can be used for differentiating as well as to identify different microsporidians with considerable accuracy.

Evaluation of the Genetic Diversity of Biovar 3 Strains of Pseudomonas syringae pv. actinidiae Isolated in Korea (RAPD 지문을 통한 우리나라에서 분리된 Pseudomonas syringae pv. actinidiae biovar 3 균주의 유전적 다양성 평가)

  • Lee, Young Sun;Kim, Gyoung Hee;Koh, Young Jin;Jung, Jae Sung
    • Journal of Life Science
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    • v.30 no.1
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    • pp.1-9
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    • 2020
  • Pseudomonas syringae pv. actinidiae, the causal agent of a bacterial canker disease in kiwifruit, is subdivided into five genetically distinct populations, namely biovars 1, 2, 3, 5, and 6. Of these, strains belonging to biovar 3 are responsible for a pandemic bacterial canker of kiwifruits since 2008. This study aimed to characterize the structure of the biovar 3 population and investigate the origin of biovar 3 strains isolated in Korea. The genetic variability of fifteen biovar 3 strains, thirteen Korean and two Chinese, were evaluated through random amplified polymorphic DNA (RAPD)-PCR. The RAPD results revealed the presence of eight lineages, designated as subgroups I-VIII, across the biovar 3 strains used in this study. As the strains in subgroups II and III from China were not found in the Korean examples, we concluded that six genetically different biovar 3 subgroups (I, IV, V, VI, VII, and VIII) are present in Korea. In PCR analysis using primers specific to the strains of New Zealand and Europe, Korean strains in subgroups V and VI amplified the relevant DNA bands, suggesting that these were introduced from these two origins, respectively. PCR primers specific to subgroup VIII were developed to monitor the spread of the first biovar 3 strain in Korea, and investigations revealed that this strain was not found in Korea after its first occurrence.

Molecular Biological Diagnosis of Meloidogyne Species Occurring in Korea

  • Oh, Hyung-Keun;Bae, Chang-Hwan;Kim, Man-Il;Wan, Xinlong;Oh, Seung-Han;Han, Yeon-Soo;Lee, Hyang-Burm;Kim, Ik-Soo
    • The Plant Pathology Journal
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    • v.25 no.3
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    • pp.247-255
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    • 2009
  • Root-knot nematode species, such as Meloidogyne hapla, M. incognita, M. arenaria, and M. javanica are the most economically notorious nematode pests, causing serious damage to a variety of crops throughout the world. In this study, DNA sequence analyses were performed on the D3 expansion segment of the 28S gene in the ribosomal DNA in an effort to characterize genetic variations in the three Meloidogyne species obtained from Korea and four species from the United States. Further, PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism), SCAR (Sequence Characterized Amplified Region) PCR and RAPD (Randomly Amplified Polymorphic DNA) were also utilized to develop methods for the accurate and rapid species identification of the root-knot nematode species. In the sequence analysis of the D3 expansion segment, only a few nucleotide sequence variations were detected among M. incognita, M. arenaria, and M, javanica, but not M. hapla. As a result of our haplotype analysis, haplotype 5 was shown to be common in M. arenaria, M. incognita, M. javanica, but not in the facultatively parthenogenetic species, M. hapla. PCR-RFLP analysis involving the amplification of the mitochondrial COII and large ribosomal RNA (lrRNA) regions yielded one distinct amplicon for M. hapla at 500 bp, thereby enabling us to distinguish M. hapla from M. incognita, M. arenaria, and M. javanica reproduced via obligate mitotic parthenogenesis. SCAR markers were used to successfully identify the four tested root-knot nematode species. Furthermore, newly attempted RAPD primers for some available root-knot nematodes also provided some species-specific amplification patterns that could also be used to distinguish among root-knot nematode species for quarantine purposes.

Variation of RAPD patterns between Male and Female Genomic DNAs in Dioecious Rumex acetosa L. (자웅이주 식물 수영 (Rumex acetosa L.)에서 암.수에 따른 RAPD pattern의 다양성 분석)

  • 김동순;구달회;허윤강;방재욱
    • Korean Journal of Plant Resources
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    • v.16 no.1
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    • pp.55-60
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    • 2003
  • The genetic variation of random amplified polymorphic DNA (RAPD) patterns of genomic DNAs was investigated in dioecious plant Rumex acetosa L., which carries different sex chromosome complements in female (2n=12A+XX) and male (2n=12A+XY$_1$Y$_2$). One hundred and twenty random primers consisted of 10-mer were used for PCR amplification. Polymorphic bands were found in 24 primers. Specific bands for female and male were 16 and 18, respectively. Especially, a band of 1,440 bp from the OPC-10 primer was male specific. These sex specific RAPD markers are used to understanding the sex determination mechanism in plants.

Genetic Diversity of Didymella bryoniae for RAPD Profiles Substantiated by SCAR Marker in Korea

  • Shim, Chang-Ki;Seo, Il-Kyo;Jee, Hyeong-Jin;Kim, Hee-Kyu
    • The Plant Pathology Journal
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    • v.22 no.1
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    • pp.36-45
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    • 2006
  • Twenty isolates of Didymella bryoniae were isolated from infected cucurbit plants in various growing areas of southern Korea in 2001 and 2002. Random Amplified Polymorphic DNA (RAPD) group [RG] I of D. bryoniae was more virulent than RG IV to watermelon. Virulence of the RG I isolate was strong to moderate to cucumber, whereas that of the RG IV varied from strong, moderate to weak. Two hundred seventy-three amplified fragments were produced with 40 primers, and were analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYSPC. At the distance level of 0.7, two major genomic DNA RAPD groups were differentiated among 20 isolates. The RG I included 7 isolates from watermelon and one isolate from melon, whereas the RG IV included 12 isolates from squash, cucumber, watermelon and melon. Amplification of internal transcribed spacer (ITS) region and small subunit rRNA region from the 20 isolates yielded respectively a single fragment. Restriction pattern with 12 restriction enzymes was identical for all isolates tested, suggesting that variation in the ITS and small subunit within the D. bryoniae were low. Amplification of the genomic DNAs of the tested isolates with the sequence characterized amplified regions (SCAR) primer RG IF-RG IR specific for RG I group resulted in a single band of 650bp fragment for 8 isolates out of the 20 isolates. Therefore, these 8 isolates could be assigned into RG I. The same experiments done with RG IIF-RG IIR resulted in no amplified PCR product for the 20 isolates tested. An about 1.4 kb-fragment amplified from the RG IV isolates was specifically hybridized with PCR fragments amplified from genomic DNAs of the RG IV isolates only, suggesting that this PCR product could be used for discriminating the RG IV isolates from the RG I isolates as well other fungal species.

Genetic Polymorphism of Marsh Clam (Corbicula leana) Identified by RAPD- PCR

  • Yoon Jong-Man;Park Kwan-Ha;Choe Sun-Nam
    • Fisheries and Aquatic Sciences
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    • v.6 no.1
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    • pp.13-19
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    • 2003
  • Genomic DNA from the muscle of marsh clam (Corbicula leana) from Gochang, Muan and a Chinese site was extracted to identify genetic differences and similarity by randomly amplified polymorphic DNAs-polymerase chain reaction (RAPD- PCR). Out of 20 primers, seven primers produced amplified fragments which were consistently polymorphic. A total of 1,246 amplified products were produced of which 530 were polymorphic $(42.5\%)$. The number of polymorphic bands produced per primer varied from 40 to 122 with an average of 75.7 in marsh clam from Gochang. 3.28 of the 23.0 polymorphic bands per lane were found to be polymorphic. Also, about $4.34\%$ of total polymorphic bands were specific to marsh clam from Gochang. The major common bands of 0.28 kb generated by primer OPB-15 (GGAGGGTGTT) were present in every individuals, which were polymorphic. This common bands in every individuals should be diagnostic of specific strains, species and/or their relatedness. Primer OPB-19 (ACCCCCGAAG) produced the highest number of 12 specific bands. The intra-population variation was revealed in the band patterns identified by this primer. The random primer OPB-12 (CCTTGACGCA) yielded the amplified fragments which were consistently polymorphic between the marsh clams from Gochang and from Muan. This primer produced a total of 77 polymorphic bands: 31 bands from Gochang, 14 from Muan and 32 from the Chinese populations. An average of polymorphic bands were 1.8 from Gochang and 2.5 from the Chinese populations. This value obtained from the Chinese population was higher than those from the two domestic populations. Generally, the RAPD polymorphism generated by these primers may be useful as a genetic marker for strain or population identification of marsh clam.

Use of RAPD Fingerprinting for Discriminating Two Populations of Hilsa shad (Tenualosa ilisha Ham.) from Inland Rivers of Bangladesh

  • Shifat, Rehnuma;Begum, Anwara;Khan, Haseena
    • BMB Reports
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    • v.36 no.5
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    • pp.462-467
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    • 2003
  • The Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) was applied to analyze the genetic variation of the Hilsa shad, Tenualosa ilisha Ham., from the two major inland rivers (Padma and Meghna) in Bangladesh. Twenty-eight random 10-mer primers were primarily scored in 8 individuals from each of the two locations. Fifteen primers, which gave polymorphism, were selected and used in the final analysis of 34 individuals from the two sites. Using these primers, 480 scorable DNA fragments were found, of which 98 (20.41%) were polymorphic. By comparing the RAPD banding patterns, variations were found between and within the populations. A dendrogram was constructed with the polymorphic fragments to analyze the genetic distances between the Hilsa shad populations. The results show two major clusters of Padma and Meghna, assuming different spawning populations with different stocks or races of Hilsa shad in the major Bangladesh rivers.