• Journal of Life Science
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• v.16 no.5
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• pp.740-745
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• 2006

Inter simple sequence repeat (ISSR) markers were performed in order to analyse the phylogenetic relationships of four taxa of Castor-aralia (Kalopanax pictus): K. pictus, K. pictus var. magnificus, K. pictus var. maximowiczii, and thornless K. pictus. The 11 primers were produced 64 reproducible ISSR bands. Analysis of ISSR from individual plants of Korean K. pictus resulted in 41 polymorphic bands with 64.1%. When species were grouped by four taxa, within group diversity was 0.115 $(H_S)$, while among group diversity was 0.467 $(G_{ST})$ on a per locus basis. The estimated gene flow (Nm) for K. pictus var. maximowiczii and K. pictus var. magnificus were very higher than K. pictus. It is suggested that the isolation of geographical distance and reproductive isolation among K. pictus populations may have played roles in shaping the population structure of this species. In phenetic tree, ISSR markers are very effective in classifying natural populations as well as taxon levels of genus Kalopanax in Korea.

• Molecules and Cells
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• v.26 no.3
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• pp.250-257
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• 2008

Microsatellites or simple sequence repeats (SSR) are widely distributed in eukaryotic genomes and are informative genetic markers. Despite many advantages of SSR markers such as a high degree of allelic polymorphisms, co-dominant inheritance, multi-allelism, and genome-wide coverage in various plant species, they also have shortcomings such as low polymorphic rates between genetically close lines, especially in Capsicum annuum. We developed an alternative technique to SSR by normalizing and alternating anchored primers in random amplified microsatellite polymorphisms (RAMP). This technique, designated reverse random amplified microsatellite polymorphism (rRAMP), allows the detection of nucleotide variation in the 3' region flanking an SSR using normalized anchored and random primer combinations. The reproducibility and frequency of polymorphic loci in rRAMP was vigorously enhanced by translocation of the 5' anchor of repeat sequences to the 3' end position and selective use of moderate arbitrary primers. In our study, the PCR banding pattern of rRAMP was highly dependent on the frequency of repeat motifs and primer combinations with random primers. Linkage analysis showed that rRAMP markers were well scattered on an intra-specific pepper map. Based on these results, we suggest that this technique is useful for studying genetic diversity, molecular fingerprinting, and rapidly constructing molecular maps for diverse plant species.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.48 no.1
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• pp.56-64
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• 2003

If a quantitative trait loci (QTL) marker identified in a population is applicable to different populations (marker universality), this will not only reduce the labor and cost in marker assisted selection (MAS), but accelerate the application of molecular markers to real breeding programs. Present study aims to evaluate the defined QTL related markers from a population to a different breeding population for the MAS. Four rice breeding populations were subjected to seventy-five simple sequence repeat (SSR) markers which were already identified for their polymorphism information content (PIC) in the parents of the crossings. Among them, eight markers were evaluated for their correlation between presence of marker alleles and phenotypic expression in breeding populations. A reasonable level of polymorphism for the mapped markers originated from any sources of rice accessions was observed between crosses of any sources (marker repeatability). However, correlation between presence of markers and expression of the traits in rice breeding populations was not significant except for minor portion of traits and markers examined (failure of marker universality). In the present study, various strategies were discussed to develop new markers with universality of breeding application.

• Korean Journal of Medicinal Crop Science
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• v.25 no.6
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• pp.411-417
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• 2017

Background: Adenophora triphylla var. japonica (Regel) H. Hara shows vegetative growth with radical leaves during the first year and shows reproductive growth with cauline leaves and bolting during the second year. In addition, the shape of the plant varies within the same species. For this reason, there are limitations to classifying the species by visual examination. However, there is not sufficient genetic information or molecular tools to analyze the genetic diversity of the plant. Methods and Results: Approximately 34.59 Gbp of raw data containing 342,487,502 reads was obtained from next generation sequencing (NGS) and these reads were assembled into 357,211 scaffolds. A total of 84,106 simple sequence repeat (SSR) regions were identified and 14,133 primer sets were designed. From the designed primer sets, 95 were randomly selected and were applied to the genomic DNA which was extracted from five plants and pooled. Thirty-nine primer sets showing more than two bands were finally selected as SSR markers, and were used for the genetic relationship analysis. Conclusions: The 39 novel SSR markers developed in this study could be used for the genetic diversity analysis, variety identification, new variety development and molecular breeding of A. triphylla.

• Mycobiology
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• v.46 no.4
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• pp.421-428
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• 2018

The white button mushroom (Agaricus bisporus) is one of the most widely cultivated species of edible mushroom. Despite its economic importance, relatively little is known about the genetic diversity of this species. Illumina paired-end sequencing produced 43,871,558 clean reads and 69,174 contigs were generated from five offspring. These contigs were subsequently assembled into 57,594 unigenes. The unigenes were annotated with reference genome in which 6,559 unigenes were associated with clusters, indicating orthologous genes. Gene ontology classification assigned many unigenes. Based on genome data of the five offspring, 44 polymorphic simple sequence repeat (SSR) markers were developed. The major allele frequency ranged from 0.42 to 0.92. The number of genotypes and the number of alleles ranged from 1 to 4, and from 2 to 4, respectively. The observed heterozygosity and the expected heterozygosity ranged from 0.00 to 1.00, and from 0.15 to 0.64, respectively. The polymorphic information content value ranged from 0.14 to 0.57. The genetic distances and UPGMA clustering discriminated offspring strains. The SSR markers developed in this study can be applied in polymorphism analyses of button mushroom and for cultivar discrimination.

• Plant Breeding and Biotechnology
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• v.6 no.4
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• pp.321-336
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• 2018

Sesame (Sesamum indicum L.) is an important oilseed crop grown in tropical and subtropical areas. The objective of this study was to investigate the genetic relationships among 129 sesame landraces and cultivars using simple sequence repeat (SSR) markers. Out of 70 SSRs, 23 were found to be informative and produced 157 alleles. The number of alleles per locus ranged from 3 - 14, whereas polymorphic information content ranged from 0.33 - 0.86. A distance-based phylogenetic analysis revealed two major and six minor clusters. The population structure analysis using a Bayesian model-based program in STRUCTURE 2.3.4 divided 129 sesame accessions into three major populations (K = 3). Based on pairwise comparison estimates, Pop1 was observed to be genetically close to Pop2 with $F_{ST}$ value of 0.15, while Pop2 and Pop3 were genetically closest with $F_{ST}$ value of 0.08. Analysis of molecular variance revealed a high percentage of variability among individuals within populations (85.84%) than among the populations (14.16%). Similarly, a high variance was observed among the individuals within the country of origins (90.45%) than between the countries of origins. The grouping of genotypes in clusters was not related to their geographic origin indicating considerable gene flow among sesame genotypes across the selected geographic regions. The SSR markers used in the present study were able to distinguish closely linked sesame genotypes, thereby showing their usefulness in assessing the potentially important source of genetic variation. These markers can be used for future sesame varietal classification, conservation, and other breeding purposes.

• Journal of Mushroom
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• v.18 no.4
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• pp.323-330
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• 2020

Agaricus bisporus is an important edible mushroom that is used as a functional food. In this study, European A. bisporus strains were analyzed for genetic diversity, population structure, and genetic differentiation using simple sequence repeat (SSR) markers. European A. bisporus strains were divided into four groups by distance-based analysis and two subpopulations by model-based analysis. The SSR markers used in this study did not group European A. bisporus strains by geographical region or pileus color. Genetic diversity was high in Group 4 based on distance-based analysis and Pop. 2 based on model-based analysis. A. bisporus strains showed very low genetic differentiation. The results of this study can be used for breeding A. bisporus in the future.

• The Korean Journal of Mycology
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• v.42 no.2
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• pp.159-164
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• 2014

For development of a method for differentiation of Pleurotus eryngii cultivars, simple sequence repeats (SSR) from whole genomic DNA sequence analysis was used for genotyping and two multiplex-SSR primer sets were developed. These SSR primer sets were employed to distinguish 12 cultivars and strains. Five polymorphic markers were selected based on the genotyping results. PCR using each primer produced one to four distinct bands ranging in size from 200 to 300 bp. Polymorphism information content (PIC) values of the five markers were in the range of 0.6627 to 0.6848 with an average of 0.6775. Unweighted pairgroup method with arithmetic mean clustering analysis based on genetic distances using five SSR markers classified 12 cultivars into two clusters. Cluster I and II were comprised of four and eight cultivars, respectively. Two multiplex sets, Multi-1 (SSR312 and SSR366) and Multi-2 (SSR178 and SSR277) completely discriminated 12 cultivars and strains with 21 alleles and a PIC value of 0.9090. These results might be useful in providing an efficient method for the identification of P. eryngii cultivars with separate PCR reactions.

• Journal of Mushroom
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• v.14 no.4
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• pp.174-178
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• 2016

Simple sequence repeats (SSR), also referred to "microsatellites" consist of tandemly repeated short DNA sequence motifs and have been applied in various marker-based studies. SSRs were isolated and characterized from 'Heuktari' and 'Miso', which are major oyster mushroom cultivars in Korea, by genome sequencing and bioinformatic analysis. The genome sizes of 'Heuktari' and 'Miso' were estimated to be 40.8 and 40.3 Mb, respectively, which are larger than those of other P. ostreatus species (PC9 and PC10) and smaller than those of P. eryngii (KNR2312P5). In total, 949 and 968 SSRs were found in the 'Heuktari' and 'Miso' genomes, respectively. Comparative analysis of five mushrooms including P. ostreatus var. florida (PC9 and PC15) and P. eryngii revealed that the number of SSRs in 'Heuktari' and 'Miso' were the highest among them. All mushrooms studied showed similar SSR distribution patterns. Tri-, hexa-, and octanucleotide motifs accounted for the top three fractions of all SSRs.

• Molecules and Cells
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• v.39 no.2
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• pp.141-148
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• 2016

Oriental melon (Cucumis melo L. var. makuwa) is one of six subspecies of melon and is cultivated widely in East Asia, including China, Japan, and Korea. Although oriental melon is economically valuable in Asia and is genetically distinct from other subspecies, few reports of genome-scale research on oriental melon have been published. We generated 30.5 and 36.8 Gb of raw RNA sequence data from the female and male flowers, leaves, roots, and fruit of two oriental melon varieties, Korean landrace (KM) and Breeding line of NongWoo Bio Co. (NW), respectively. From the raw reads, 64,998 transcripts from KM and 100,234 transcripts from NW were de novo assembled. The assembled transcripts were used to identify molecular markers (e.g., single-nucleotide polymorphisms and simple sequence repeats), detect tissue-specific expressed genes, and construct a genetic linkage map. In total, 234 single-nucleotide polymorphisms and 25 simple sequence repeats were screened from 7,871 and 8,052 candidates, respectively, between the KM and NW varieties and used for construction of a genetic map with 94 F2 population specimens. The genetic linkage map consisted of 12 linkage groups, and 248 markers were assigned. These transcriptome and molecular marker data provide information useful for molecular breeding of oriental melon and further comparative studies of the Cucurbitaceae family.