• Horticultural Science & Technology
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• v.31 no.3
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• pp.337-343
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• 2013

The aims of this study were to compare genetic distances among 14 Phalaenopsis varieties using simple sequence repeat (SSR) and sequence-related amplified polymorphism (SRAP) marker systems and to determine the discrimination using SSR. A total of 111 SSR primers and 30 SRAP combinations were initially screened. Twelve SSR primers and thirty SRAP combinations showed high polymorphism among the 14 Phalaenopsis varieties including domestic breeding varieties, conserved in National Institute of Horticultural & Herbal Science (NIHHS). The amplified DNA fragments were separated by denaturing acrylamide gels and detected by silver staining method. A total of 474 polymorphic bands, including 55 by SSRs and 419 by SRAPs, were identified and used for genetic diversity analysis. Polymorphic bands were scored for calculating a simple matching coefficient of genetic similarity and cluster analysis with multi-variate statistical package (MVSP) 3.1. Fourteen Phalaenopsis varieties were classified into three major groups at similarity coefficient value of 0.683 and 0.66 using SRAP and SSR, respectively. Also we could discriminate these domestic breeding Palaenopsis varieties using only SSR 20 and SSR 22. The results indicate that SSR analysis is effective for discrimination among Phalaenopsis varieties and SRAP is useful for genetic diversity when there is no sequence information. These studied SSR and SRAP markers will be useful tools for genotype identification, germplasm conservation and genetic relationship study in Phalaenopsis.

• Journal of Korean Society of Forest Science
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• v.95 no.4
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• pp.435-442
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• 2006

A total of 167 peculiar haplotypes confirmed from 28 cpSR variants that were observed in 19 populations of Japanese red pine in Korea through cpSSR marker analysis. Thirteen individuals that showed identical haplotype dispersed evenly in 10 populations, and the average number of effective haplotype within population was 13.37. Estimate of genetic diversity (He) was 0.987 on the basis of cpSSR haplotype variants that was equivalent to or higher than the estimates reported in other studies on some forest tree species. Estimation of genetic diversity (S.I.) on the basis of cpSSR variants composing each haplotype revealed the highest estimate of 1.109 for the population of Gangwon-Yeongwol and the lowest estimate of 0.411 for the population of Gyeongbuk Mungyeong with the average of 0.887. Most of observed cpSSR variants appeared to exist commonly in 19 populations (97.62%), and genetic differentiation of cpSSR variants among populations was turned out to be weak (${\Phi}_{ST}=0.024$). Relatively fast rate of mutation of cpSSR marker might be a major cause for such weak population differentiation. There was no identical haplotype shared between 39 population pairs of 173 pair-wise population pairs. Estimation of genetic distance among 19 populations on the basis of population pairs was also impossible, that might be resulted from restricted migration among 19 populations. Considering the observed distribution patterns of cpSSR variants in addition to the previous studies on I-SSR variants, informations on the present geographic location and genetic status of populations should be considered together for effective sustainable management of the genetic resources of Japanese red pine in Korea.

• Horticultural Science & Technology
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• v.31 no.6
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• pp.772-781
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• 2013

The objective of this study was to develop simple sequence repeat (SSR) markers from expressed sequence tags (EST) of lettuce (Lactuca sativa) and identify 9 germplasms from 3 wild species of lettuce and 61 commercial cultivars using the developed EST-SSR markers. A total of 81,330 lettuce ESTs from NCBI databases were used to search for SSR and 4,229 SSR loci were identified. The highest proportion (59.12%, 2500) was represented by trinucleotide, followed by dinucleotide (29.70%, 1256) and hexanucleotide (6.62%, 280) among SSR repeat motifs. Totally 474 EST-SSR primers were developed from EST and a random set of 267 primers was used to assess the genetic diversity among 9 germplasms and 61 cultivars. Out of 267 primers, 47 EST-SSR markers showed polymorphism between 7 cultivars. Twenty-six EST-SSR markers among 47 EST-SSR markers showed high polymorphism, reproducibility, and band clearance. The relationship between 26 markers genotypes and 70 accessions was analyzed. Totally 127 polymorphic amplified fragments were obtained by 26 EST-SSR markers and two to nine SSR alleles were detected for each locus with an average of 4.88 alleles per locus. Average polymorphism information content was 0.542, ranging from 0.269 to 0.768. Genetic distance of clusters ranged from 0.05 to 0.94 between 70 accessions and dendrogram at a similarity of 0.34 gave 7 main clusters. Analysis of genetic diversity revealed by these 26 EST-SSR markers showed that the 9 germplasms and 61 commercial cultivars were discriminated by marker genotypes. These newly developed EST-SSR markers will be useful for cultivar identification and distinctness, uniformity and stability test of lettuce.

• Genomics & Informatics
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• v.9 no.4
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• pp.189-193
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• 2011

Simple sequence repeats (SSRs) are ubiquitous short tandem duplications found within eukaryotic genomes. Their length variability and abundance throughout the genome has led them to be widely used as molecular markers for crop-breeding programs, facilitating the use of marker-assisted selection as well as estimation of genetic population structure. Here, we report a software application, "SSR-Primer Generator " for SSR discovery, SSR-primer design, and homology-based search of in silico amplicons from a DNA sequence dataset. On submission of multiple FASTA-format DNA sequences, those analyses are batch processed in a Java runtime environment (JRE) platform, in a pipeline, and the resulting data are visualized in HTML tabular format. This application will be a useful tool for reducing the time and costs associated with the development and application of SSR markers.

• Horticultural Science & Technology
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• v.32 no.6
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• pp.845-852
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• 2014

Among various abiotic stress factors, soil salinity decreases the photosynthetic rate, growth, and yield of plants. Recently, many genes have been reported to enhance salt tolerance. The objective of this study was to characterize the Brassica rapa Salt Stress Resistance (BrSSR) gene, of which the function was unclear, although the full-length sequence was known. To characterize the role of BrSSR, a B. rapa Chinese cabbage inbred line ('CT001') was transformed with pSL94 vector containing the full length BrSSR cDNA. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis showed that the expression of BrSSR in the transgenic line was 2.59-fold higher than that in the wild type. Analysis of phenotypic characteristics showed that plants overexpressing BrSSR were resistant to salinity stress and showed normal growth. Microarray analysis of BrSSR over-expressing plants confirmed that BrSSR was strongly associated with ERD15 (AT2G41430), a gene encoding a protein containing a PAM2 motif (AT4G14270), and GABA-T (AT3G22200), all of which have been associated with salt tolerance, in the co-expression network of genes related to salt stress. The results of this study indicate that BrSSR plays an important role in plant growth and tolerance to salinity.

• 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.

• Horticultural Science & Technology
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• v.29 no.4
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• pp.366-373
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• 2011

To analyze the genetic relationships among Korean potato cultivars and to develop cultivar identification method using DNA markers, we carried out genotyping using simple sequence repeats (SSR) analysis and developed multiplex-SSR set. Initially, we designed 92 SSR primer combinations reported previously and applied them to twenty four Korean potato cultivars. Among the 92 SSR markers, we selected 14 SSR markers based on polymorphism information contents (PIC) values. PIC values of the selected 14 markers ranged from 0.48 to 0.89 with an average of 0.76. PIC value of PSSR-29 was the lowest with 0.48 and PSSR-191 was the highest with 0.89. UPGMA clustering analysis based on genetic distances using 14 SSR markers classified 21 potato cultivars into 2 clusters. Cluster I and II included 16 and 5 cultivars, respectively. And 3 cultivars were not classified into major cluster group I and II. These 14 SSR markers generated a total of 121 alleles and the average number of alleles per SSR marker was 10.8 with a range from 3 to 34. Among the selected markers, we combined three SSR markers, PSSR-17, PSSR-24 and PSSR-24, as a multiplex-SSR set. This multiplex-SSR set used in the study can distinguish all the cultivars with one time PCR and PAGE (Polyacrylamide gel electrophoresis) analysis and PIC value of multiplex-SSR set was 0.95.

• Proceedings of the KAIS Fall Conference
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• 2009.12a
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• pp.501-504
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• 2009

Sympathetic skin response (SSR) is defined as a minute change of skin potential after electrical stimulation. This test measures the change in voltage that originates from the surface of the skin and is attributed to sudomotor activity. The aim of this study was to define the criteria for validation of the responses. 40 normal subjects (20-73 years of age) with non-sympathetic dysfunction were tested and SSR was generated form all subjects. SSR latency was 1331.22${\pm}$177.51ms in the right palm, 1331.74${\pm}$156.42ms in the left palm, 1851.79${\pm}$220.99ms in the right sole, and 1874.10${\pm}$215.01ms in the left sole. And SSR amplitude was 595.83${\pm}$221.16${\mu}$V in the right palm, 605.33${\pm}$226.45${\mu}$V in the left palm, 291.76${\pm}$133.36${\mu}$V in the right sole, and 288.77${\pm}$129.70${\mu}$ V in the left sole. SSR latency and amplitude had no significantly difference between the right and the left side. SSR latency was consistently shorter (p<0.001) and SSR amplitude higher (p<0.001) in feet than in hands. SSR waveforms were P-type (32 subjects, 75%) and N-type (8 subjects, 25%), respectively. The SSR latency and amplitude in palms/soles were closely correlated with age (p<0.05) and height (p<0.05). The SSR test is one of methods assessing impairment of sympathetic fibers in peripheral neuropathy as well as a disorder of sympathetic system in other diseases and so our results from normal healthy subjects can be used as clinical criteria for SSR test.

• Horticultural Science & Technology
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• v.31 no.6
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• pp.748-755
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• 2013

This study was conducted to find a salt tolerance gene in Brassica rapa. In order to meet this objective, we analyzed data from a KBGP-24K oligo chip [BrEMD (Brassica rapa EST and microarray database)] of the B. rapa ssp. pekinensis 'Chiifu' under salt stress (250 mM NaCl). From the B. rapa KBGP-24K microarray chip analysis, 202 salt-responsive unigenes were primarily selected under salt stress. Of these, a gene with unknown function but known full-length sequence was chosen to closely investigate the gene function. The selected gene was named BrSSR (B. rapa salt stress resistance). BrSSR contains a 285 bp open reading frame encoding a putative 94-amino acid protein, and a DUF581 domain. The pSL94 vector was designed to over-express BrSSR, and was used to transform tobacco plants for salt tolerance analysis. T1 transgenic tobacco plants that over-expressed BrSSR were selected by PCR and DNA blot analyses. Quantitative real-time RT PCR revealed that the expression of BrSSR in transgenic tobacco plants increased by approximately 3.8-fold. Similar results were obtained by RNA blot analysis. Phenotypic characteristics analysis showed that transgenic tobacco plants with over-expressed BrSSR were more salt-tolerant than the wild type control under 250 mM NaCl for 5 days. Based on these results, we hypothesized that the over-expression of BrSSR may be closely related to the enhancement of salt tolerance.

• Journal of Korean Society of Forest Science
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• v.102 no.1
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• pp.59-65
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• 2013

The jujube is an important fruit tree species in Korea. Traditionally, classifications of jujube cultivars have been based on morphological characters; however, morphological identification can be problematic because morphological traits are affected by environmental conditions. Therefore, DNA markers are now being used for the rapid and accurate identification of plant species. Inter-simple sequence repeat (I-SSR) is one of the best DNA-based molecular marker techniques, which is useful for studying genetic relations and for the identification of closely related cultivars. In this study, 5 Korean jujube trees and 1 jujube tree imported from China were analyzed for 16 I-SSR primers. Amplification of the genomic DNA of jujube cultivars by using I-SSR analysis generated 100 bands, with an average of 6.25 bands per primer, of which 45 bands (45%) were polymorphic. The number of amplified fragments with I-SSR primers ranged from 2 to 13. The percentage of polymorphism ranged from 10% to 100%. I-SSR finger printing profiles showed that 'Boeun jujube' and 'Daeri jujube' had characteristic DNA patterns, indicating unequivocal cultivar identification at molecular level. According to the results of clustering analysis, the genetic similarity coefficient ranged from 0.68 to 0.92. 'Boeun jujube' and 'Daeri jujube' were divided into independent groups, and 'Bokjo jujube', 'Geumseong jujube', 'Wolchul jujube', and 'Mudeung jujube' were placed in the same group. Therefore, I-SSR markers are suitable for the discrimination of 'Boeun jujube' and 'Daeri jujube' cultivars.