• Title/Summary/Keyword: RAPD primer analysis

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Distinction between Cold-sensitive and -tolerant Jute by DNA Polymorphisms

  • Hossain, Mohammad Belayat;Awal, Aleya;Rahman, Mohammad Aminur;Haque, Samiul;Khan, Haseena
    • BMB Reports
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    • v.36 no.5
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    • pp.427-432
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    • 2003
  • Jute is the principal coarse fiber for commercial production and use in Bangladesh. Therefore, the development of a high-yielding and environmental-stress tolerant jute variety would be beneficial for the agro economy of Bangladesh. Two molecular fingerprinting techniques, random-amplified polymorphic DNA (RAPD) and amplified-fragment length polymorphism (AFLP) were applied on six jute samples. Two of them were cold-sensitive varieties and the remaining four were cold-tolerant accessions. RAPD and AFLP fingerprints were employed to generate polymorphism between the cold-sensitive varieties and cold-tolerant accessions because of their simplicity, and also because there is no available sequence information on jute. RAPD data were obtained by using 30 arbitrary oligonucleotide primers. Five primers were found to give polymorphism between the varieties that were tested. AFLP fingerprints were generated using 25 combinations of selective-amplification primers. Eight primer combinations gave the best results with 93 polymorphic fragments, and they were able to discriminate the two cold-sensitive and four cold-tolerant jute populations. A cluster analysis, based on the RAPD and AFLP fingerprint data, showed the population-specific grouping of individuals. This information could be useful later in marker-aided selection between the cold-sensitive varieties and cold-tolerant jute accessions.

Induction of Valiant of Cyrtomium caryoptideum var. coreanum Nakai by Chemical Mutagenesis In vitro and RAPD Analysis (기내에서 화학돌연변이원 처리에 의한 참쇠고비의 변이주 유기 및 RAPD 분석)

  • Jeong Jin-A;Lee Cheol-Hee
    • Korean Journal of Plant Resources
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    • v.19 no.2
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    • pp.374-380
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    • 2006
  • With the aim of inducing mutation in fern Cyrtomium caryoptideum var. coreanum, rhizome segments of In vitro-grown cultures were treated with chemical mutagens such as EMS, NMU and colchicine. Based on regeneration ratios, sensitivities for each treatments were assessed and also optimum treatment condition of each mutagens was explored. Optimum concentration for EMS treatment was considered to be 20 to 50mM and for NMU 5 to 10mM. NMU was found to be more effective in inducing chlorophyll and morphological variations than EMS. The RAPD were performed to check the genetic modification of phenotypical variants. As a result, polymorphic DNA band patterns between wild type and variants were observed by two 10-mer primers.

DNA Profiles Analysed by Polymerase Chain Reaction-Randorn Amplified Polymorphic DNAs in Shortnecked Clam (Ruditapes philippinarum) Populations

  • Yoon, Jong-Man;Kim, Yong-Ho;Kim, Jong-Yeon
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2002.05a
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    • pp.281-282
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    • 2002
  • Out of 20 primers, 6 generated a total of 1,111 major and minor RAPD bands, producing approximately 4.2 average polymorphic bands per primer in shortnecked clam (Ruditapes philippinarum) population from Anmyeondo. The Bandsharing value altered from 0.15 to 0.74, with the average f 0.51, as calculated by bandsharing analysis. The RAPD profiles obtained with DNAs of two populations from Anmyeondo and Seocheon, respectively, were considerably different (0.20 and 0.51, respectively). The varying degrees of difference among populations amy also be of relevance to the restricted hybridization of wild bivalve. Besides gene mapping and breeding applications, PCR-RAPD systems could be very useful for the rapid certification and quality control of seed production and for every projects based on PCR amplification of specific bivalve DNA fragments.

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DNA Profiles Analysed by Polymerase Chain Reaction-Random Amplified Polymorphic DNAs in Shortnecked Clam (Ruditapes philippinarum) Populations

  • Yoon, Jong-Man;Kim, Yong-Ho;Kim, Jong-Yeon
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2002.08a
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    • pp.172-174
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    • 2002
  • Out of 20 primers, 6 generated a total of 1,11 major and minor RAPD bands, producing approximately 4.2 average polymorphic bands pe primer in shortnecked clam (Ruditapes philippinarum) population from Anmyeondo. The bandsharing value altered form 0.15 to 0.74, with the average of 0.5, as calculated by bandsharing analysis. The RAPD profiles obtained with DNAs of two populations from Anmyeondo and Seocheon, respectively, were considerably different (0.20 and 0.51, respectively). The varying degrees of difference among populations may also be of relevance to the retricted hybridization of wild bivalve. Besides gene mapping and breeding applications, PCR-RAPD system could be very useful for the rapid certification and quality control of seed production and for every projects based on PCR amplification of specific bivalve DNA fragments.

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Analysis of Lactobacillus casei and Mutant Strains by Polymerase Chain Reaction (Polymerase Chain Reaction에 의한 Lactobacillus casei 및 돌연변이 균주들의 비교 분석)

  • Nam, Jin-Sik;Lee, Jeong-Jun;Shin, Myeong-Su;Na, Seog-Hwan;Baek, Young-Jin;Yoo, Min
    • Microbiology and Biotechnology Letters
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    • v.22 no.6
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    • pp.577-583
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    • 1994
  • To classify Lactobacillus casei strains on the basis of difference in their chromosomal DNA sequence, we have performed polymerase chain reactions on their chromosomal DNA by using random primers, and followed by analyzing randomly amplified polymorphic DNA fragments. We also developed a mini-preparative method to isolate PCR-grade chromosomal DNA from Lacto- bacillus casei strains within 3 hours. Based on RAPD pattems by polymerase chain reactions with degenerated random primers, 4 Lactobacillus casei strains and 2 mutant strains were successfully discriminated. Results were very sensitive, strain-specific and reproducible. It was also reliable. These results suggest that RAPD may be applied efficiently for the identification of several Lactoba- cillus casei strains.

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Bandsharing Values and Genetic Distances of Two Wild Shortnecked Clam, Ruditapes philippinarum Populations from the Yellow Sea Assessed by Random Amplified Polymorphic DNAs-Polymerase Chain Reaction

  • Yoon, Jong-Man;Kim, Yong-Ho
    • Journal of Aquaculture
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    • v.17 no.1
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    • pp.12-23
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    • 2004
  • Genomic DNAs were extracted from the muscle of twenty-two specimens of two shortnecked clam, Ruditapes phifippinarum populations collected in Anmyeondo and Seocheon. Genetic differences within and between populations were analysed by random amplified polymorphic DNAs-polymerase chain reaction (RAPD-PCR) using twenty arbitrary decamer primers. Out of 20 primers, 6 generated a total of 1,111 major and minor RAPD bands from individuals of two sites, producing approximately 4.2 average polymorphic bands per primer in individuals from Anmyeondo and ranging in size from less than 50 to larger than 1,500 base pairs (bp). The electrophoretic analysis of RAPD products amplified showed moderate levels of similarity among the different individuals in Seo-cheon population. The average bandsharing values (BS value) of the samples within population from Anmyeondo ranged from 0.155 to 0.684, whereas it was 0.143∼0.782 within population from Seocheon. The average BS value between individuals No. 13 and No. 14 from Seocheon was 0.782 which was higher than that of those from Anmyeondo. The single linkage dendrogram resulted from three primers (OPA-08, -09 and -20), indicating six genetic groupings composed of group 1 (No.4, 8 and 10), group 2 (No. 18), group 3 (No.2, 5 and 7), group 4 (No. 1, 3, 6, 9, 11, 12, 13, 14, 15 and 17), group 5 (16, 19 and 20) and group 6 (No. 21 and 22). In the Seocheon population, the individual No. 18 clustered distinctly from the others of this population. The observed genetic distance between the two populations from Anmyeondo and Seocheon was more than 0.209 (0.247 and 0.275). The shortest genetic distance (0.094) displaying significant molecular differences was between individuals No. 13 and No. 14. Especially, the genetic distance between individuals No. 22 and the remnants among individuals in two geographical populations was highest (0.275). This result illustrated that individual No.22 is distinct from other individuals within two shortnecked populations. The different geographical features of two sites may have caused the genetic diversity in two shortnecked clam populations.

The Discrimination of Coisis Semen and Coisis lacrima-jobi Semen by the Random Amplified Polymorphic DNAs and Anatomical Characteristics (의이인과 염주의 RAPD분석 및 해부학적 특징에 의한 감별)

  • Lee, Mi-Young;Im, Seung-Hi;Kim, Ho-Kyoung;Han, Keong-Sik;Choi, Yong-Hyu;Ju, Young-Seung;Oh, Seung-Eun;Ko, Byoung-Seob
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.1
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    • pp.17-23
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    • 2002
  • The seeds of Coix lachryma-jobi Linne var. mayuen Stapf. are used as dietary food for obesity and diabetes under the names of Yulmu in Korea and Yiyiren(薏苡仁) in China. It is one of the drugs promoting diuresis to eliminate the wetness-evil from the lower warmer in the traditional Korean medicine. According to ancient textbook of the traditional Korean medicine, it should be applied to patients with phlegm and heat, etc. The establishment of the method for the discrimination of Coisis Semen is very important for the quality control of drugs. Random amplified polymorphic DNA(RAPD) analysis and anatomical characteristics were used for the discrimination of Coix lachryma-jobi $Linn\acute{e}$ var. mayuen $S_{TAPF}$. and C. lachryma-jobi $Linn\acute{e}$. In the RAPD analysis with 20 primers, 8 primers gave informative and reproducible bands with the genomic DNA. From the cluster analysis, the genus Coix were divided into two groups at similarity coefficient of 0.863.

Analysis of Genetic Diversity in Thirteen Turfgrass Cultivars Cultivated at Golf Courses Using RAPD Markers (RAPD마커를 이용한 국내골프장의 잔디 13 품종의 유전적 다양성 분석)

  • Kim, Min-Jeong;Kim, Tae-Soo;Shim, Chang-Ki;Kim, Yong-Ki;Jee, Hyeong-Jin
    • Weed & Turfgrass Science
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    • v.1 no.4
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    • pp.57-63
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    • 2012
  • This study was carried our to examine the genetic relationship of 13 commercial turfgrass cultivars using Random Amplified Polymorphic DNA to provide genetic informations more efficient golf course management. Analysis of 56 random hexamer primers generated 13 to 54 polymorphic bands among the 13 cultivars with an average of 30.7 bands per primer. The results of cluster analysis based on RAPDs revealed that three major variety groups: Group I - 'Shadow II', 'Aurora Gold', 'Little Bighorn Blue', 'PennA-1', and 'PennA-4'; Group II - 'Midnight II', 'Prosperity', 'Moon light SLT', 'Bright star SLT', and 'Silver dollar'; and Group III - 'Olympic Gold', 'Silver Star', and 'Tar Heel II'. The genetic similarity coefficients among 13 turfgrass cultivars ranged from 0.039 to 1.0 with highest coefficient in Group III. Studies on morphological characters and the effective molecular markers such as sequence characterized amplified regions are further needed to identify relationships and genetic diversities within species and among species.

Genetic Diversity of Rehmannia glutinosa Genotypes Assessed by Molecular Markers (분자표지자에 의한 지황 유전집단의 유전적 다양성)

  • Bang, Kyong-Hwan;Chung, Jong-Wook;Kim, Young-Chang;Lee, Jei-Wan;Kim, Hong-Sig;Kim, Dong-Hwi
    • Journal of Life Science
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    • v.18 no.4
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    • pp.435-440
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    • 2008
  • Random amplified polymorphic DNA (RAPD) markers were used to identify the genetic diversities among and within varieties and landraces of Rehmannia glutinosa. Polymorphic and reproducible bands were produced by 10 primers out of total 20 primers used in the experiment. In RAPD analysis of the 11 genotypes, 64 fragments out of 73 amplified genomic DNA fragments were polymorphic which represented an average 6.4 polymorphic fragments per primer. Number of amplified fragments with random primers ranged from 2 (OPA-1) to 13 (OPA-11) and varied in size from 200 bp to 1,400 bp. Especially, OPA-10, OPA-11 and OPA-19 primers showed specific bands for varieties of Korea Jiwhang and Jiwhang il ho, which could be useful for discriminating from other varieties and landraces of R. glutinosa. Percentage polymorphism ranged from a minimum of 50% (OPA-1) to a maximum of 100% (OPA-11), with an average of 87.7%. Similarity coefficients were higher in the genotypes of Korea Jiwhang and Jiwhang il ho than in other populations. In cluster analysis, genotypes of Korea Jiwhang, Jiwhang il ho, and Japanese accession were separated from those of other varieties and landraces. Average of genetic diversity within the population $(H_S)$ was 0.110, while average of total genetic diversity $(H_T)$ was 0.229. Across all RAPD makers the $G_{ST}$ value was 0.517, indicating that about 52% of the total genetic variation could be explained by RAPDs differences while the remaining 48% might be attributable to differences among samples. Consequently, RAPD analysis was useful method to discriminate different populations such as domestic varieties and other landraces. The results of the present study will be used to understand the population and evolutionary genetics of R. gllutinosa.

Phylogenetic Relationships and Genetic Diversity in Collected Resources of Carthamus tinctorius by Random Amplified Polymorphic DNA Markers (RAPD 마커에 의한 수집된 홍화자원에서 계통관계와 유전적 다양성)

  • Sung, Jung-Sook;Cho, Gyu-Taek;Lee, Gi-An;Baek, Hyung-Jin;Huh, Man-Kyu
    • Journal of Life Science
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    • v.20 no.12
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    • pp.1764-1771
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    • 2010
  • Carthamus tinctorius L. (Compositae) is an herb primarily distributed throughout in the world. The species is regarded as ecologically important in the world. Safflower was used for medicines, as well as making red (carthamin) and yellow dyes. We have used the RAPD technique to investigate the phylogenetic relationships and genetic diversity of C. tinctorius. We obtained 123 bands from all the 26 cultivars. The average number of bands was 9.5 per primer. The genetic diversity of safflower is found among cultivars and there is a high among-cultivar differentiation. The OPC18-01 band is the specific marker for Syria cultivar, whereas no products were detected in individuals from other country cultivars. We found seven phenetic bands for determining the specific marker of cultivars with SCAR markers. Though the number of individuals sampled for analysis was small and probably not fully representative of the total available diversity in C. tinctorius, this study demonstrates that the regions (Morocco, Syria, and Turkey) of the Mediterranean Sea were more variable than other regions with the exception of India. In this result, although only simple result of RAPD is difficult to assert the center of species diversity of C. tinctorius, the regions of the Mediterranean Sea may be the most probable candidate for the origin of safflower. India was also the candidate of the center or secondary center of species diversity of C. tinctorius. RAPD markers were effective in classifying cultivar levels of safflower.