• Title/Summary/Keyword: Polymorphic Loci

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Chromosome-specific polymorphic SSR markers in tropical eucalypt species using low coverage whole genome sequences: systematic characterization and validation

  • Patturaj, Maheswari;Munusamy, Aiswarya;Kannan, Nithishkumar;Kandasamy, Ulaganathan;Ramasamy, Yasodha
    • Genomics & Informatics
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    • v.19 no.3
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    • pp.33.1-33.10
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    • 2021
  • Eucalyptus is one of the major plantation species with wide variety of industrial uses. Polymorphic and informative simple sequence repeats (SSRs) have broad range of applications in genetic analysis. In this study, two individuals of Eucalyptus tereticornis (ET217 and ET86), one individual each from E. camaldulensis (EC17) and E. grandis (EG9) were subjected to whole genome resequencing. Low coverage (10×) genome sequencing was used to find polymorphic SSRs between the individuals. Average number of SSR loci identified was 95,513 and the density of SSRs per Mb was from 157.39 in EG9 to 155.08 in EC17. Among all the SSRs detected, the most abundant repeat motifs were di-nucleotide (59.6%-62.5%), followed by tri- (23.7%-27.2%), tetra- (5.2%-5.6%), penta- (5.0%-5.3%), and hexa-nucleotide (2.7%-2.9%). The predominant SSR motif units were AG/CT and AAG/TTC. Computational genome analysis predicted the SSR length variations between the individuals and identified the gene functions of SSR containing sequences. Selected subset of polymorphic markers was validated in a full-sib family of eucalypts. Additionally, genome-wide characterization of single nucleotide polymorphisms, InDels and transcriptional regulators were carried out. These variations will find their utility in genome-wide association studies as well as understanding of molecular mechanisms involved in key economic traits. The genomic resources generated in this study would provide an impetus to integrate genomics in marker-trait associations and breeding of tropical eucalypts.

Genetic Variation of Several Isoenzymes in Pinus densiflora for. multicaulis (반송(Pinus densiflora for. multicaulis)의 몇 가지 동위효소(同位酵素)의 유전변이(遺傳變異))

  • Hwang, Jae Woo;Lee, Seok Woo
    • Journal of Korean Society of Forest Science
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    • v.85 no.3
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    • pp.409-415
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    • 1996
  • Isozyme variations in 4 enzyme systems in Pinus densiflora for. multicaulis in 31 individual trees collected throughout the country were studied by starch gel electrophoresis using haploid megagametophyte tissue to compare with those of Pinus densiflora. A minimum of 7 loci were found to code for isozymes of the enzyme systems. No variation was found at locus GOT-A; at the remaining 6 variable loci(GDH-A, GOT-B, GOT-C, IDH-A, LAP-,A, LAP-B) and 2 to 4 alleles were identified. We could not find any marker alleles to distinguish Pinus densiflora for. multicaulis from Pinus densiflora at the isozymes studied here. Allele frequency distributions at each loci were almost all the same as those of P. densiflora. The percentage of polymorphic loci(99% level), the number of alleles per locus, the observed and expected heterozygosities were 85.7, 2.3, 0.165 and 0.186%, respectively. The level of genetic diversity in Pinus densiflora for. multicaulis seemed to be less than that of P. densiflora.

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Study of Genetic Diversity among Simmental Cross Cattle in West Sumatra Based on Microsatellite Markers

  • Agung, Paskah Partogi;Saputra, Ferdy;Septian, Wike Andre;Lusiana, Lusiana;Zein, Moch. Syamsul Arifin;Sulandari, Sri;Anwar, Saiful;Wulandari, Ari Sulistyo;Said, Syahruddin;Tappa, Baharuddin
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.2
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    • pp.176-183
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    • 2016
  • A study was conducted to assess the genetic diversity among Simmental Cross cattle in West Sumatra using microsatellite DNA markers. A total of 176 individual cattle blood samples was used for obtaining DNA samples. Twelve primers of microsatellite loci as recommended by FAO were used to identify the genetic diversity of the Simmental Cross cattle population. Multiplex DNA fragment analysis method was used for allele identification. All the microsatellite loci in this study were highly polymorphic and all of the identified alleles were able to classify the cattle population into several groups based on their genetic distance. The heterozygosity values of microsatellite loci in this study ranged from 0.556 to 0.782. The polymorphism information content (PIC) value of the 12 observed loci is high (PIC>0.5). The highest PIC value in the Simmental cattle population was 0.893 (locus TGLA53), while the lowest value was 0.529 (locus BM1818). Based on the genetic distance value, the subpopulation of the Simmental Cross-Agam and the Simmental Cross-Limapuluh Kota was exceptionally close to the Simmental Purebred thus indicating that a grading-up process has taken place with the Simmental Purebred. In view of the advantages possessed by the Simmental Cross cattle and the evaluation of the genetic diversity results, a number of subpopulations in this study can be considered as the initial (base) population for the Simmental Cross cattle breeding programs in West Sumatra, Indonesia.

The Genetic Diversity of Trans-caucasian Native Sheep Breeds

  • Hirbo, Jibril;Muigai, Anne;Naqvi, A.N.;Rege, E.D.;Hanotte, Olivier
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.7
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    • pp.943-952
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    • 2006
  • The genetic variation in 10 indigenous Caucasian sheep breeds was studied with 14 micro-satellite loci in order to determine the genetic diversity among and between the breeds. Five breeds from Asia, five breeds from Europe and one breed from Africa, were included in order to study any relationships or influences they may have with the Caucasian sheep analyzed. A Karakul population from Uzbekistan was included in the study to see whether there was any Central Asian influence. All the 14 loci were found to be polymorphic in all the breeds, with the exception of ILST0056, which was monomorphic in Imeretian. A total of 231 alleles were generated from all the 688 individuals of the sheep analyzed. The mean number of alleles (MNA) at each locus was 16.5. The total number of alleles detected in all samples ranged from 13 in several loci to 23 in OarJMP029. Out of total 308 Hardy-Weinberg Equilibrium (HWE) tests, 85 gave significant results. After Bonferroni correction for multiple tests, 30 comparisons still remained significant to the experimental levels. The Gala population was the most diverse and Imeretian the least diverse with a MNA of 8.50 and 5.51, respectively. Gene diversity estimates exhibited the same trend and ranged from 0.803 in Gala and 0.623 in Imeretian, but generally there is higher diversity among the Caucasian breeds in comparison to other eference breeds. The closest breeds were Tushin and Bozakh with Da of 0.113 and most distant breeds were $Djallonk{\acute{e}}$ and North Rondalsy with Da of 0.445. Principal Component (PC) analyses were done. PC1 described 14% of the differences. PC2, which described 13% of the differences, further separated the Caucasian breeds from Asian breeds except Karakul and Awasi, and the two British breeds. PC3 described 10% of the differences, allowing better differentiation of the Caucasian breeds. A moderate degree of reliability was observed for individual-breed assignment from the 14 loci using different approaches among which the Bayesian method proved to be the most efficient. About 72% of individuals analyzed were correctly assigned to their respective breeds.

Development and characterization of microsatellite markers for an endangered species, $Epinephelus$ $bruneus$, to establish a conservation program

  • An, Hye-Suck;Kim, Jae-Woo;Lee, Jang-Wook;Kim, Shin-Kwon;Lee, Bae-Ik;Kim, Dae-Jung;Kim, Yi-Cheong
    • Animal cells and systems
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    • v.16 no.1
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    • pp.50-56
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    • 2012
  • Kelp grouper ($Epinephelus$ $bruneus$ Bloch 1793) is a commercially important fish in Korea. In recent years, the catch of kelp grouper in the coastal waters of Korea has significantly declined. Despite its importance, little is known about its genetic diversity and conservation efforts are hampered. In this study, we isolated and characterized 12 microsatellite loci using an enrichment method based on magnetic/biotin capture of microsatellite sequences from a size-selected genomic library. All loci were readily amplified and contained TG/CA denucleotide repeats. To characterize each locus, 30 individuals from a natural E. bruneus population in the coastal waters of Jeju Island, Korea, were genotyped. All loci except three, KEm118, KEm154, and KEm219, were polymorphic, with an average of 8.1 alleles per locus (range 2-18). The mean observed and expected heterozygosities were 0.47 (range 0.19-1.00) and 0.61 (range 0.29-0.92), respectively. A significant deviation from Hardy-Weinberg equilibrium was observed at three loci (KEm134, KEm184, and KEm283). These findings will be useful for effective monitoring and management of genetic variation of kelp grouper as well as for the implementation of a fisheries conservation program.

Ethnic Differences in Allelic Frequencies of Two (CA)n Microsatellite Markers Located on Chromosome 5q

  • Hong, Sung-Soo;Chae, Jae-Jin;Goh, Sung-Ho;Yong, Koong-Nam;Lee, Chung-Choo
    • Animal cells and systems
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    • v.1 no.1
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    • pp.123-128
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    • 1997
  • The characteristics of allelic polymorphisms of the two (CA)n microsatellite (p599 and ㅅ599) markers spanning the long arm of chromosome 5 were studied in 52 DNA samples from unrelated inhabitants of Seoul (Korea) by using the polymerase chain reaction (PCR) to investigate differences in allele frequencies between Korean and Caucasian populations. The 6 alleles were observed for p599 (CA)n with a polymorphism informative content (PIC) value of 0.71 and 9 alleles for ㅅ599 (CA)n with a PIC value of 0.82. The observed heterozygote frequencies of the loci were estimated to 0.730 and 0.846, respectively. Several allele frequencies of two loci showed significant differences between Korean and Caucasian populations. Genotype data from the two loci were consistent with the Hardy-Weinberg equilibrium by x2 test. Linkage disequilibrium between p599 (CA)n and ㅅ599 (CA)n loci was observed in x2 test between the observed and expected frequency of allelic association. The probability of matching calculated at each locus was 0.104 for p599 (CA)n and 0.043 for ㅅ599 (CA)n, respectively. These results demonstrate the need to determine populationspecific allele frequency distributions for polymorphic markers when performing genetic linkage studies in racially defined several populations.

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Development of a High-Resolution Multi-Locus Microsatellite Typing Method for Colletotrichum gloeosporioides

  • Mehta, Nikita;Hagen, Ferry;Aamir, Sadaf;Singh, Sanjay K.;Baghela, Abhishek
    • Mycobiology
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    • v.45 no.4
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    • pp.401-408
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    • 2017
  • Colletotrichum gloeosporioides is an economically important fungal pathogen causing substantial yield losses indifferent host plants. To understand the genetic diversity and molecular epidemiology of this fungus, we have developed a novel, high-resolution multi-locus microsatellite typing (MLMT) method. Bioinformatic analysis of C. gloeosporioides unannotated genome sequence yielded eight potential microsatellite loci, of which five, CG1 $(GT)_n$, CG2 $(GT1)_n$, CG3 $(TC)_n$, CG4 $(CT)_n$, and CG5 $(CT1)_n$ were selected for further study based on their universal amplification potential, reproducibility, and repeat number polymorphism. The selected microsatellites were used to analyze 31 strains of C. gloeosporioides isolated from 20 different host plants from India. All microsatellite loci were found to be polymorphic, and the approximate fragment sizes of microsatellite loci CG1, CG2, CG3, CG4, and CG5 were in ranges of 213-241, 197-227, 231-265, 209-275, and 132-188, respectively. Among the 31 isolates, 55 different genotypes were identified. The Simpson's index of diversity (D) values for the individual locus ranged from 0.79 to 0.92, with the D value of all combined five microsatellite loci being 0.99. Microsatellite data analysis revealed that isolates from Ocimum sanctum, Capsicum annuum (chili pepper), and Mangifera indica (mango) formed distinct clusters, therefore exhibited some level of correlation between certain genotypes and host. The developed MLMT method would be a powerful tool for studying the genetic diversity and any possible genotype-host correlation in C. gloeosporioides.

Genetic Diversity and Phenetic Relationships of Genus Oxalis in Korea Using Random Amplified Polymorphic DNA (RAPD) Markers (RAPD마크를 이용한 한국 내 괭이밥속 식물의 유전적 다양성과 표현형 관계)

  • Huh, Man Kyu;Choi, Byoung-Ki
    • Journal of Life Science
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    • v.24 no.7
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    • pp.707-712
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    • 2014
  • We evaluated the phenetic relationships within six taxa of genus Oxalis L. in Korea with random amplified polymorphic DNA (RAPD) markers. Ten primers produced 125 bands for six taxa, and the mean number of bands per primer was 12.5. Across the six taxa, 121 (96.8%) bands were polymorphic, and only four were monomorphic. The mean number of RAPD phenotypes across the six taxa varied from 3.6 (O. stricta and O. corymbosa) to 4.8 (O. corniculata for. rubrifolia). In a simple measure of intraspecies variability according to the percentage of polymorphic bands, O. stricta and O. corymbosa exhibited the lowest variation (28.8%), and O. corniculata for. rubrifolia showed the highest (38.4%). A mean of 32.7% of the loci was polymorphic within taxa. The total interspecies genetic diversity ($H_T$) and intraspecies genetic diversity ($H_S$) was 0.362 and 0.122, respectively. On a per-locus basis, the proportion of total genetic variation due to differences among species ($G_{ST}$) was 0.663. This indicates that about 66.3% of the total variation was among species. The node of O. stricta and O. corniculata for. rubrifolia was strongly supported, with a high bootstrap value in the NJ tree and sistered with O. corniculata. According to RAPD analysis, the number of chromosomes was not congruent with a phenetic relationship.

Establishment of an Individual Identification System Based on Microsatellite Polymorphisms in Korean Cattle (Hanwoo)

  • Yoon, Du-Hak;Kong, Hong-Sik;Oh, Jae-Don;Lee, Jun-Heon;Cho, Byung-Wook;Kim, Jong-Dae;Jeon, Ki-Jun;Jo, Chang-Yun;Jeon, Gwang-Joo;Lee, Hak-Kyo
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.6
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    • pp.762-766
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    • 2005
  • This study was conducted to establish an individual identification system comprising of 19 microsatellite markers located on different bovine autosomes. The markers were typed on 257 animals from five cattle breeds. In total, 112 alleles were detected from the genotyping of 19 microsatellite markers. The average heterozygosities ranged from 0.292 to 0.824 and the polymorphic information content (PIC) ranged from 0.274 to 0.817 in Hanwoo. We found that there were differences in allele frequencies in Hanwoo when compared with other cattle breeds. The calculated cumulative power of discrimination (CPD) was 99.999% when nine microsatellite loci were used for analysis in the individual identification system. Also the matching probability, the probability that two unrelated animals would show the same genotypes, was estimated to be $0.44{\times}10^{-9}$. Therefore, the nine markers used in this study will be used for individual identification in two million Hanwoo individuals.

Assessment of genetic diversity and distance of three Cicuta virosa populations in South Korea

  • Nam, Bo Eun;Kim, Jae Geun;Shin, Cha Jeong
    • Journal of Ecology and Environment
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    • v.36 no.3
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    • pp.205-210
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    • 2013
  • Cicuta virosa L. (Apiaceae) is a perennial emergent plant designated as an endangered species in South Korea. According to the former records, only four natural habitats remain in South Korea. A former study suggested that three of four populations (Pyeongchang: PC, Hoengseong: HS, Gunsan: GS) would be classified as different ecotypes based on their different morphological characteristics and life cycle under different environmental conditions. To evaluate this suggestion, we estimated genetic diversity in each population and distance among three populations by random amplification of polymorphic DNA. Seven random primers generated a total of 61 different banding positions, 36 (59%) of them were polymorphic. Nei's gene diversity and the Shannon diversity index increased in the order of PC < HS < GS, which is the same order of population size. In the two-dimensional (2D) plot of first two principal components in principal component analysis with the presence of 61 loci, individuals could be grouped as three populations easily (proportion of variance = 0.6125). Nei's genetic distance for the three populations showed the same tendency with the geographical distance within three populations. And it is also similar to the result of discriminant analysis with the morphological or life-cycle factors from the previous study. From the results, we concluded that three different populations of C. virosa should be classified as ecotypes based on not only morphology and phenology but genetic differences in terms of diversity and distance as well.