• Title/Summary/Keyword: Polymorphic markers

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Genetic Variability Based on Randomly Amplified Polymorphic DNA in Kacip Fatimah (Labisia pumila Benth & Hook f) collected from Melaka and Negeri Sembilan States of Malaysia

  • Bhore, Subhash J.;Nurul, A.H.;Shah, Farida H.
    • Journal of Forest and Environmental Science
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    • v.25 no.2
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    • pp.93-100
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    • 2009
  • In Malaysia, Labisia pumila Benth & Hook f, popularly known as 'Kacip Fatimah' has been used traditionally to treat various elements of the woman's health in Malay community. The objective of this study was to develop randomly amplified polymorphic DNA (RAPD) based DNA markers for the identification of L. pumila and to distinguish its three varieties from each other. Total DNA from nine accessions of L. pumila was extracted by CTAB method and polymerase chain reactions (PCR) were carried out to amplify the segments of DNA using different primers to develop DNA barcode using RAPD technique. To find out variety-specific DNA marker/s, twenty different 10-mer primer sequences with annealing temperature from 36-$40^{\circ}C$ were evaluated in triplicate. Out of 20 random primers, two primers (OPA-1 and OPA-2/A10) were selected which produced reliable RAPD band patterns. To have DNA based handle, two RAPD amplification products were cloned and sequenced to determine the identity of the DNA. RAPD analysis using two random primers generated 72 discrete bands ranging in size 200 bp-3,000 bp. Fifty nine of these were polymorphic loci (82%) and thirteen were non-polymorphic loci (18%). A total of 32 bands polymorphic loci (72%) were amplified with primer OPA-1 and analyzed by cluster analysis and UPGMA (Unweighted Pair Group Method with Arithmetic) to present a dendogram depicting the degree of genetic relationship among nine accessions of L. pumila. Our results shows the reasonable genetic diversity among the L. pumila varieties and within varieties; and two RAPD marker sequences obtained could be used to identify L. pumila at species level.

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Genetic diversity analysis of fourteen geese breeds based on microsatellite genotyping technique

  • Moniem, Hebatallah Abdel;Zong, Yang Yao;Abdallah, Alwasella;Chen, Guo-hong
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.11
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    • pp.1664-1672
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    • 2019
  • Objective: This study aimed to measure genetic diversity and to determine the relationships among fourteen goose breeds. Methods: Microsatellite markers were isolated from the genomic DNA of geese based on previous literature. The DNA segments, including short tandem repeats, were tested for their diversity among fourteen populations of geese. The diversity was tested on both breeds and loci level and by mean of unweighted pair group method with arithmetic mean and structure program, phylogenetic tree and population structure were tested. Results: A total of 108 distinct alleles (1%) were observed across the fourteen breeds, with 36 out of the 108 alleles (33.2%) being unique to only one breed. Genetic parameters were measured per the 14 breeds and the 9 loci. Medium to high heterozygosity was reported with high effective numbers of alleles (Ne). Polymorphic information contents (PIC) of the screened loci was found to be highly polymorphic for eleven breeds; while 3 breeds were reported moderately polymorphic. Breeding coefficient ($F_{IS}$) ranged from -0.033 to 0.358, and the pair wise genetic differentiation ($F_{ST}$) ranged from 0.01 to 0.36 across the fourteen breeds; for the 9 loci observed and expected heterozygosity, and Ne were same as the breeds parameters, PIC of the screened loci reported 6 loci highly polymorphic and 3 loci to be medium polymorphic, and $F_{IS}$ ranged from -0.113 to 0.368. In addition, genetic distance estimate revealed a close genetic distance between Canada goose and Hortobagy goose breeds by 0.04, and the highest distance was between Taihu goose and Graylag goose (anser anser) breed by 0.54. Conclusion: Cluster analyses were made, and they revealed that goose breeds had hybridized frequently, resulting in a loss of genetic distinctiveness for some breeds.

Studies on Genetic Variation of Different Chinese Duck Populations with Random Amplified Polymorphic DNA Analysis

  • Su, Y.;Liu, C.W.;Liu, L.;Ye, C.H.;Cao, W.Q.;Huang, Y.Q.;Zheng, J.;Cai, D.Y.;Olowofeso, O.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.4
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    • pp.475-481
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    • 2006
  • The genetic polymorphism and relationships of Muscovy, Cherry Valley Meat ducks, Partridge ducks and their crossbreds $F_1$ and $F_2$, respectively, were studied using a random amplified polymorphic DNA (RAPD) technique. The results showed that RAPD markers were effective for the analysis of genetic relationships among ducks. Amplification with 20-primers gave 760 reproducible amplified fragments. The percentage of polymorphic marker band was 74.70%, which indicates that the RAPD technique had higher efficiency of polymorphism detection and sensitivity in studying the genetic variations among ducks and showed that the genetic polymorphism was abundant between two species of duck populations. The average index of genetic distance in hybrid $F_2$ was 0.2341 and higher than that of its parents, which indicates that the genetic diversity was improved by crossbreeding with Muscovy.

Genetic Diversity and Population Structure of Comus controversa Hemsley Using RAPD (RAPD에 의한 층층나무의 유전적 다양성과 집단구조)

  • Moon, Sung-Gi;Huh, Man-Kyu
    • Journal of Life Science
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    • v.18 no.2
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    • pp.175-179
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    • 2008
  • Cornus controversa is a long-lived woody species mostly distributed in East Asia. Random amplified polymorphic DNA (RAPD) markers were used to investigate the genetic diversity and population structure of Korean populations of this species. A high level of genetic variation was found in seven populations of C. controversa. The mean genetic diversity (H) was 0.222 across populations, varying from 0.200 to 0.238. Eighty of the 93 loci (86.0%) showed detectable polymorphism in at least one population. Total genetic diversity values ($H_T$) varied between 0.192 and 0.231, giving an average overall polymorphic loci of 0.212. The interlocus variation of genetic diversity within populations ($H_S$) was high (0.167). Mean of genetic diversity in C. controversa was higher than average values for species with similar life history traits. The sexual reproduction, perennial habitat, and longevity are proposed as possible factors contributing to high genetic diversity. On a per locus basis, the proportion of total genetic variation due to differences among populations ($G_{ST}$) ranged from 0.169 to 0.278 with a mean of 0.216, indicating that about 21.6% of the total genetic variation was among populations. An indirect estimate of the number of migrants per generation (Nm=1.893) indicated that gene flow was extensive among Korean populations of C. controversa.

Genetic Diversity and DNA Polymorphism in Platycodon grandiflorum DC. Collected from East-Asian Area

  • Park, Chun-Geun;Yan, Zhi-Yi;Lee, Sang-Chul;Shon, Tae-Kwon;Park, Hee-Woon;Jin, Dong-Chun
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.2
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    • pp.115-120
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    • 2005
  • Broadening the genetic base of Platycodon grandiflorum DC. cultivar to sustain improvement requires assessment of genetic diversity available in P. grandiflorum DC.. The objective of this study was to analyze the genetic variation, genetic relationship among 48 samples collected from East-Asian Area by means of RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction) markers. From the 18 primers tested, produced total 211 bands with an average of 11.7 bands per primer and obtained 103 polymorphic band with an average of 5.7 bands per primer,s revealed relatively high percentage of polymorphic bands (48.8%). The genetic similarities calculated from RAPD data varied from 0.688 to 0.994 and were clustered to six major groups on a criterion of 0.78 similarity coefficient. The present study has revealed the significant genetic similarity among the samples tested. The analysis of genetic relationships in P. grandiflorum using RAPD-PCR banding data can be useful for the breed improvement.

Development of microsatellite markers for Hosta capitata (Asparagaceae) and amplification in related taxa

  • CHOI, Mi-Jung;LEE, Jung-Hyun;CHO, Won-Bum;HAN, Eun-Kyeong;CHOI, Hyeok-Jae
    • Korean Journal of Plant Taxonomy
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    • v.50 no.3
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    • pp.327-332
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    • 2020
  • Microsatellite markers were developed as a tool for phylogeographic studies of Hosta capitata. We also assessed cross-amplification in species closely related to Hosta capitata. We produced 28 polymorphic microsatellite markers by mapping 300 bp paired-end reads obtained from Illumina MiSeq data of H. capitata. In H. capitata, the number of alleles per locus ranged from 1 to 13. Observed and expected heterozygosity ranged from 0.000 to 0.844 and 0.000 to 0.832, respectively. Additionally, 13 loci were successfully transferable to the related species of H. minor and H. venusta. These markers will provide a powerful genetic tool not only for elucidating the phylogeographic patterns of H. capitata populations but also for studying the genetic delimitation of H. capitata from its related species.

Construction of genetic linkage maps of Allium cepa using genotyping-by-sequencing

  • Lee, Daewoong;Chung, Yong Suk;Kim, Changsoo;Jun, Tae-Hwan
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.117-117
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    • 2017
  • The onion (Allium cepa L.) is the most widely cultivated species of the genus Allium, especially it has been valued because of the pungent flavor and aroma. Allium species including onion has very large genome sizes ranging from approximately 10 to 20 Gbp, which have complicated genomic studies and precluded genome sequencing until recently. A population of 186 F2 individuals derived from a cross of 'Umjinara' ${\times}$ 'Sinsunhwang' and the two parental lines were used for this study. For the development of framework map, various types of markers including SSRs, RAPD, SNPs, and CAPS makers have been used for polymorphism test. Especially, a lot of SNP and CAPS loci were developed from the onion transcriptome sequence by RNASEQ of two parental lines. The GBS libraries have been constructed based on a modified protocol from Poland Lab using a two-enzyme system. We have been developing markers showing polymorphism between two parental lines, and genotyping for all F2 individuals were finished for a number of polymorphic markers. For the construction of GBS libraries, a set of 192 barcoded adapters were generated from complementary oligonucleotides with XhoI overhang sequence and unique barcodes of length 4-8 bp and they have been tested using two parental linesto determine the optimum conditions for GBS analysis.

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Genetic Diversity of 14 Indigenous Grey Goose Breeds in China Based on Microsatellite Markers

  • Tu, Yunjie;Chen, K.W.;Zhang, S.J.;Tang, Q.P.;Gao, Y.S.;Yang, N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.1
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    • pp.1-6
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    • 2006
  • This experiment first cloned some microsatellite sequences for goose species by magnetic beads enriched method and studied the genetic structure research of 14 indigenous grey goose breeds using 19 developed and 12 searched microsatellite markers with middle polymorphism. According to the allele frequencies of 31 microsatellite sites, mean heterozygosity (H), polymorphism information content (PIC) and $D_A$ genetic distances were calculated for 31-microsatellite sites. The results showed that 25 of 31microsatellite sites were middle polymorphic, so the 25 microsatellite markers were effective markers for analysis of genetic relationship among goose breeds. The mean heterozygosity was between 0.4985 and 0.6916. The highest was in the Xupu (0.6916), and in the Yan was the lowest (0.4985) which was consistent with that of PIC. The phylogenetic tree was completed through analysis of UPGMA. Fencheng Grey, Shoutou, Yangjiang and Magang were grouped firstly, then Xongguo Grey, Wugang Tong, Changle and Youjiang were the second group; Gang, Yan Xupu and Yili were the third group; Yongkang Grey and Wuzeng were the fourth group. The results could provide basic molecular data for the research on the characteristics of local breeds in the eastern China, and a scientific basis for the conservation and utilization of those breeds.

Identification of RAPD Markers Associated with Grain Weight in Rice

  • Lee, Hyung-Gyu;Kim, Kyung-Min;Sohn, Jae-Keun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.4
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    • pp.261-265
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    • 2001
  • This study was carried out to select randomly amplified polymorphic DNA (RAPD) markers associated with grain weight of a large-grain mutant, Hyacp 39-26-1, derived from anther culture of a rice cultivar, 'Hwayeongbyeo'. The segregation mode for grain weight in an F$_2$ population from a cross, 'Hwayeongbyeo/Hyacp 39-26-1', showed a nearly normal distribution. One hundred and ninety-one F$_2$plants ranged from 21.8 g to 34.7 g in 1,000-grain weight with a mean of 26.8 g. Five hundred and twenty primers were used to detect the RAPD markers associated with the grain weight of the large-grain mutant. Of these primers, 54 primers showed polymorphism between 'Hwayeongbyeo' and 'Hyacp 39-26-1'. Four RAPD markers (OPB18, OPH07, OPT20, and OPX20) were significantly related to the grain weight of twenty one F$_3$ lines derived from the cross, 'Hwayeongbyeo/Hyacp 39-26-1'. This RAPD marker could facilitate the early and efficient selection of high-yield lines through improvement of grain weight in rice.

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Second locus for late-onset familial Amyotrophic Lateral Sclerosis (가족성 근위축성측삭경화증을 유발시키는 두 번째 유전자 위치)

  • 홍성출
    • Journal of Life Science
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    • v.11 no.3
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    • pp.279-283
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    • 2001
  • Amyotrophic lateral sclerosis(ALS) is a progressive neurologic disorder resulting from the degeneration of upper and lower motor neurons, and is inherited in 10% of cases. About 20% of familial ALS, clinically indistinguishable from sporadic ALS, is caused by mutations of Cu/Zn superoxide dismutase on chromosome 21q22.21 inherited as an autosomal dominant trait. We now report a new locus in the non-SOD1 dominantly inherited ALS. We screened a large ALS family with 11 affected individuals and one obligate gene carrier with genome-wide ABI polymorphic markers using the ABI 377 automated system. No evidence of linkage was obtained with the autosomal markers. We next screened this family with X chromosome markers as there was no evidence of male-to-male tran-smission of the disease. Linkage was established with several X chromosome markers with a lod score up to 3.8; almost the maximum possible score in this family. Our finding imply that a gene for the dominant expression of a neuronal degeneration is coded on X chromosome and raise the question of the role of X-linked genes that escape inactivation in this pathogenesis. More importantly, our finding that a gene causing ALS is localized on X-chromosome has direct investigational relevance to sporadic ALS, where epidemiological studies show male gender predominance(1.3:1) and earlier onset in men by 5-10 years.

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