• Title/Summary/Keyword: RAPD primer analysis

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Molecular Characterization of Seaweeds Using RAPD and Differential Display

  • HONG Yong-Ki;KIM Yong-Tae;KIM Se-Kwon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.6
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    • pp.770-778
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    • 1996
  • A rapid and economical method of simultaneous extraction of DNA and RNA from seaweeds has been developed by the use of lithium chloride. Lithium chloride facilitates the softening of cell walls resulting in a decrease in both compressive and tensile modulus of elasticity. The DNA was characterized by high molecular weight larger than 27 kb and a relative lack of carbohydrate and protein contamination. The DNA and RNA extracted by the method from many seaweeds were of sufficient quality to be used as a template for per amplification with a plant intergenic gene primer set, for RAPD analysis with arbitrary primers, and for differential display with arbitrary primers in the morphologically distinct regions of the matured Porphyra thallus. The cDNA polymorphism indicated that the reproductive tissue types (male, female, patch) had a relatively high degree of similarity; the vegetative tissue types (dividing, non-dividing) also showed a similar pattern with respect to each other. Holdfast tissue had very low similarity with the other tissues, but appeared most similar to vegetative non-dividing tissue type.

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Genetic Relationships of Lactuca spp. Revealed by RAPD, Inter-SSR, AFLP, and PCR-RFLP Analyses

  • Yang, Tae-Jin;Jang, Suk-Woo;Kim, Won-Bae
    • Journal of Crop Science and Biotechnology
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    • v.10 no.1
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    • pp.27-32
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    • 2007
  • RAPD, Inter-SSR, and AFLP markers were used to assess the genetic diversity of lettuce cultivars and the phylogenetic relationships in Lactuca spp. A total of 216 polymorphic bands from seven RAPD primers, four Inter-SSR primers, and five AFLP primer combinations were used to elucidate the genetic similarity among lettuce cultivars. Forty-four lettuce accessions were subdivided into discrete branches according to plant type: crisphead, butterhead, and stem type, with some exceptions. The leafy- and cos-type accessions were intermingled in other groups with no discrete branch indicating that these are more diverse than others. Three accessions, including the Korean cultivar 'Cheongchima', the Korean local landrace 'Jinjam', and the German cultivar 'Lolla Rossa' were classified as the most diverse accessions. Twenty bands were unique in specific cultivars. Among these, three were specific in a plant type; one in Korean leafy type, one in crisphead type, and one in cos type lettuce. In the phylogenetic analysis among Lactuca species, L. saligna, L. serriola, and L. georgica clustered in a sister branch of the L. sativa complex. Two L. virosa accessions show the highest intra-specific relationships. L. perennis outlied from all the other Lactuca species at a genetic similarity of 0.53 and clustered with two Cichorium species, C. intybus and C. endivia, with genetic similarity of 0.67. The phylogenetic tree was supported by data from polymorphism of chloroplast genome which was revealed by PCR-RFLP.

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Genetic Relationships Analysis of Asparagus cochinchinensis $L_{OUR}$ Collections by Random Amplified Polymorphic DNA (RAPD에 의한 천문동 수집종의 유연관계 분석)

  • Kang, Chan-Ho;Park, Chun-Bong;Choi, Joung-Sik;Choi, Yeong-Geun
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.5
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    • pp.384-391
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    • 2002
  • To analyze the genetic relationships among 23 accessions of Asparagus cochinensis $L_{OUR}$, random amplified polymorphic DNA(RAPD) analysis was performed using artificially synthesized 10 primers. The range of polymorphism was $42.9{\sim}91.7%$ with an average of 72.9% in 85 randomly and specifically amplified DNA fragments. On the. basis of similarity coefficient analysis by unweighed pairgroup method, arithmetic average method(UPGMA), 23 accessions of Asparagus cochinensis $L_{OUR}$ could be classified into 6 groups at the similarity coefficient value of 0.82. Group I contained 5 accessions, Group II contained 6 accessions, Group III contained 6 accessions, GroupIV contained 2 accessions, Group V contained 2 accessions and Group VI contained 2 accessions. The range of total genetic similarity coefficient value of 23 accessions of Asparagus cochinchinensis $L_{OUR}$ was $0.47{\sim}0.92$ and average value was 0.76. To obtain more exact data from PCR, we also tried to develope enhanced RAPD techniques using Bovine Lacto Transfer Technique Optimizer(BLOTTO). In RAPD analysis of Asparagus cochinensis $L_{OUR}$, we could obtain better RAPD results by adding BLOTTO at a final concentration of 1%.

Identification of Molecular Markers Linked to Ti Locus in Soybean

  • Kim Myung Sik;Park Min Jung;Hwang Jung Gyu;Jo Soo Ho;Ko Mi Suk;Chung Jong Il
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.5
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    • pp.419-422
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    • 2004
  • Soybean is a major source of protein meal in the world. Kunitz trypsin inhibitor (KTI) protein is responsible for the inferior nutritional quality of unheated or incompletely heated soybean meal. The objective of this research was to identify RAPD markers linked to KTI protein allele using bulked segregant analysis. Cultivar Jinpumkong2 (TiTi) was crossed with C242 (titi, absence of KTI protein) and F. seeds were planted. The $F_1$. plants were grown in the greenhouse to produce $F_2$ seeds. Each $F_2$ seed from $F_1$. plants was analysed electrophoretically to determine the presence of the KTI protein band. The present and absent bulks contained twenty individuals each, which were selected on the basis of the KTI protein electrophoresis, respectively. Total 94 $F_2$ individuals were constructed and 1,000 Operon random primers were used to identify RAPD primers linked to the Ti locus. The presence of KTI protein is dominant to the lack of a KTI protein and Kunitz trypsin inhibit protein band is controlled by a single locus. Four RAPD primers (OPAC12, OPAR15, OPO12, and OPC08) were linked to the Ti locus. RAPD primer OPO12 was linked to Ti locus, controlling kunitz trypsin inhibitor protein at a distance of 16.0 cM. This results may assist in study of developing fine map including Ti locus in soybean.

Mycelial Propagation and Molecular Phylogenetic Relationships of Commercially Cultivated Agrocybe cylindracea based on ITS Sequences and RAPD

  • Alam, Nuhu;Kim, Jeong-Hwa;Shim, Mi-Ja;Lee, U-Youn;Lee, Tae-Soo
    • Mycobiology
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    • v.38 no.2
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    • pp.89-96
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    • 2010
  • This study evaluated the optimal vegetative growth conditions and molecular phylogenetic relationships of eleven strains of Agrocybe cylindracea collected from different ecological regions of Korea, China and Taiwan. The optimal temperature and pH for mycelial growth were observed at $25^{\circ}C$ and 6. Potato dextrose agar and Hennerberg were the favorable media for vegetative growth, whereas glucose tryptone was unfavorable. Dextrin, maltose, and fructose were the most effective carbon sources. The most suitable nitrogen sources were arginine and glycine, whereas methionine, alanine, histidine, and urea were least effective for the mycelial propagation of A. cylindracea. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The sequence of ITS2 was more variable than that of ITS1, while the 5.8S sequences were identical. The reciprocal homologies of the ITS sequences ranged from 98 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) using 20 arbitrary primers. Fifteen primers efficiently amplified the genomic DNA. The average number of polymorphic bands observed per primer was 3.8. The numbers of amplified bands varied based on the primers and strains, with polymorphic fragments ranging from 0.1 to 2.9 kb. The results of RAPD analysis were similar to the ITS region sequences. The results revealed that RAPD and ITS techniques were well suited for detecting the genetic diversity of all A. cylindracea strains tested.

Identification and Genetic Diversity of Korean Tomato Cultivars by RAPD Markers (한국 내 토마토 재재종의 RAPD에 의한 동정과 유전적 다양성)

  • Huh, Man-Kyu;Youn, Sun-Joo;Kang, Sun-Chul
    • Journal of Life Science
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    • v.21 no.1
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    • pp.15-21
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    • 2011
  • Cultivated tomato, Lycopersicum esculentum, is a very important crop. We selected 36 cultivars and studied them for identification and polymorphism by employing random amplified DNA (RAPD) analysis with 80 oligonucleotide primers. Of the 80 primers, 36 primers (45.0%) were polymorphic. Detection of polymorphism in cultivated tomato opens up the possibility of development of its molecular map by judicious selection of genotypes. Molecular markers can also be used for cultivar identification and protection of the plant breeder's intellectual property rights (plant breeders' rights, PBRs). As an example, DNA polymorphism using OPC-13 primer that did not produce the OPC-13-01 band was only found in Junk Pink and Ailsa Craighp cultivars. OPA-12-03 and OPB-15-07 were fragments specific to the TK-70 cultivar and were absent in other cultivars. DNA polymorphism in cultivated tomato in this study was correlated with a type of inflorescence, although some cultivars had exceptions. These approaches will be useful for developing marker-assisted selection tools for genetic enhancement of the tomato plant for desirable traits.

A STUDY ON THE MYCELIA CUL TURE AND GENETIC CHARACTERISTICS IN Lepista nuda (민자주방망이버섯(Lepista nuda)의 균사배양 및 유전적 특성에 관한 연구)

  • 김종봉;황성구
    • Journal of Life Science
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    • v.11 no.5
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    • pp.496-501
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    • 2001
  • This study was carried out to investigate the cultural characteristics and the genetic characteristics in Lepists nuda. The mycelia cultural characteristics which include the specificity for pH, carbon source, nitrogen source and medium were studied using petridish culture and liquid media culture respectively. The genetic characteristics were also investigated using random amplified polymorphic DNA (RAPD) analysis. To find out the mycelial growth rate by various medium, mycelia was cultured for 25 days in PDA, YM, MEA, GPB and Yamanaka. The Yamanaka was superior to the other media in supporting the mycelial growth. pH6 produced the best result in the test of an optimal pH. In experiment for optimal carbon source, starch showa 46.8$\pm$1.7mm of diameter of mycelial colony as the best and for nitrogen source, yeast extract showed a good effect as well. In investigation for genetic characteristics, Lepists nuda was amplified by 7 primers among 10 primers and Pleurotus ostreatus was also amplified by 7 primers. From the RAPD analysis between species, the band patterns were different.

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Analysis of Morphological and Genetic Relationships amomg Isolates of the Artificially Cultivated Mushroom, Hypsizygusmarmoreus (느티만가닥버섯 균주의 형태 및 유전적 유연관계 분석)

  • Kim, Min-Kyung
    • The Korean Journal of Mycology
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    • v.48 no.3
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    • pp.313-323
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    • 2020
  • To investigate the morphological characteristics and genetic relationships among isolate of the artificially cultivated mushroom Hypsizygus marmoreus, 111 isolates were collected from Korea and other countries. Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) and ITS rDNA sequencing were used to confirm the genetic relationships among the collected H. marmoreus isolates. As a result of RAPD analysis using universal rice primer (URP)-PCR, all isolates of H. marmoreus clustered into three groups, which showed high sequence similarity (>90%). In addition, isolates with morphological and geographical differences formed independent clusters. However, it was impossible to distinguish between brown and white strains. Sixteen strains showing morphological and geographic differences were selected, and their ITS region sequences (640 bp) were aligned and compared. The ITS region sequences belonging to these isolates showed 94.8-99.1% similarities to those of publicly available H. marmoreus strains in GenBank. In conclusion, there were differences among isolates in terms of morphology and the area from which they were collected, but all the isolates used in the experiment were classified as H. marmoreus.

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.

Assessment of Bandsharing Values in RAPD-PCR Analysis of Dwarf Cattle of Kerala

  • Suprabha, P.;Anilkumar, K.;Aravindakshan, T.V.;Raghunandanan, K.V.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.9
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    • pp.1217-1220
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    • 2005
  • Random amplified polymorphic DNA (RAPD-PCR) analysis of 56 animals of four different genetic groups of dwarf cattle in Kerala was done as a single step analysis. Bandsharing (BS) values were calculated for animals of each group and between groups as an analytical tool to find out genetic variation among animals. The different factors affecting BS values were estimated using Harvey''s Least squares analysis. The effects of genetic group, Guanine-cytosine (GC) content of primer and gel on BS values were found significant. Bandsharing values of Kasargode-Highrange dwarf animals were significantly different from Vechur, Vatakara and their combinations. The Vechur, Vatakara and Vechur-Vatakara combinations were found to be more uniform (high BS value) compared with other combinations. The bandsharing value was lowest with primers of GC content 90% and highest with 80% GC content. The effect of gel on BS value points to the need of adjustments of gel factor for calculation of BS values.