• Title/Summary/Keyword: gDNA-SSR

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Characterization and Gene Co-expression Network Analysis of a Salt Tolerance-related Gene, BrSSR, in Brassica rapa (배추에서 염 저항성 관련 유전자, BrSSR의 기능 검정 및 발현 네트워크 분석)

  • Yu, Jae-Gyeong;Lee, Gi-Ho;Park, Ji-Hyun;Park, Young-Doo
    • 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.

Molecular Characterization of Hypernodulation in Soybean

  • Van, Kyu-Jung;Ha, Bo-Keun;Hwang, Eun-Young;Kim, Moon-Young;Heu, Sung-Gi;Lee, Suk-Ha
    • The Plant Pathology Journal
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    • v.19 no.1
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    • pp.24-29
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    • 2003
  • SS2-2, a hypernodulating soybean mutant was isolated by EMS mutagenesis from Sinpaldalkong 2. This auto-regulation mutant showed greater number of nodules and smaller plant size than its wild type Sinpaldalkong 2. SSR markers were used to identify DNA variation at SSR loci from different soybean LG. The only SSR marker that detected a length polymorphism between SS2-2 and its wild type ancestor was Satt294 on LG C1 instead of LG H, locating a hypernodulating gene. Sequencing data of flanking Satt294 indicated that the size variation was due to extra stretch of TTA repeats of the SSR motif in SS2-2, along with $A\longrightarrow$G transversion. In spite of phenotypic differences between the wild type and its hypernodulating mutants, genomic DNA poly-morphisms at microsatellite loci could not control regulation of nodule formation. The cDNA-AFLP method was applied to compare differential display of cDNA between Sinpaldalkong 2 and SS2-2. After isolation and sequence comparison with many AELP fragments, several interesting genes were identified. Northern blot analysis, immunolocalization and/or the yeast two-hybrid system with these genes might provide information on regulation of nodule development in SS2-2.

Development and Molecular Characterization of Novel Polymorphic Genomic DNA SSR Markers in Lentinula edodes

  • Moon, Suyun;Lee, Hwa-Yong;Shim, Donghwan;Kim, Myungkil;Ka, Kang-Hyeon;Ryoo, Rhim;Ko, Han-Gyu;Koo, Chang-Duck;Chung, Jong-Wook;Ryu, Hojin
    • Mycobiology
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    • v.45 no.2
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    • pp.105-109
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    • 2017
  • Sixteen genomic DNA simple sequence repeat (SSR) markers of Lentinula edodes were developed from 205 SSR motifs present in 46.1-Mb long L. edodes genome sequences. The number of alleles ranged from 3-14 and the major allele frequency was distributed from 0.17-0.96. The values of observed and expected heterozygosity ranged from 0.00-0.76 and 0.07-0.90, respectively. The polymorphic information content value ranged from 0.07-0.89. A dendrogram, based on 16 SSR markers clustered by the paired hierarchical clustering' method, showed that 33 shiitake cultivars could be divided into three major groups and successfully identified. These SSR markers will contribute to the efficient breeding of this species by providing diversity in shiitake varieties. Furthermore, the genomic information covered by the markers can provide a valuable resource for genetic linkage map construction, molecular mapping, and marker-assisted selection in the shiitake mushroom.

Development of SSR markers for genetic mapping of Korean ginseng and authentication of Korean ginseng cultivars

  • Kim, Nam-Hoon;Choi, Hong-Il;Jung, Ju-Yeon;Choi, Beom-Soon;Ahn, In-Ok;Lee, Joon-Soo;Yang, Tae-Jin
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.10a
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    • pp.11-11
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    • 2010
  • The Korean ginseng, Panax ginseng C. A. Meyer is a popular medicinal herb in Araliaceae. Genetic map in crops provides valuable information for breeding, genetic and genomic researches. However, little information is available for construction of genetic map in ginseng. Up to now, we have produced large amounts of expressed sequence tags (ESTs) from four ginseng cultivars (37Mb, 49Mb, 39Mb, 47Mb from Gopoong, Gumpoong, Chunpoong and Yunpoong respectively using pyrosequencing technique and 5Mb from normalized full-length cDNA library of Chunpoong) to obtain comprehensive information of gene expression, and constructed EST database including ESTs from public database. Till now, we designed 261 SSR primer sets using EST sequences and identified 106 intergenic polymorphic markers. And 44 of the 106 showed polymorphisms among panax ginseng cultivars. Among 44 markers, 27 SSR polymorphic markers were inspected to 51 $F_2$ population from Yunpoong x Chunpoong, which showed good at the fitness of Mendellian segregation ratio 1:2:1. To enrich the number of markers, and thus construct high resolution genetic map which can be used as frame map for further genome sequencing. we are planning to develop large scale EST-derived SNP markers which are available in the F2 population. This study provides genetic information as well as foundation for ginseng researches such as genetics, genomics, breeding, and the final goal for whole genome sequencing. This study was supported by Technology Development Program for Agriculture and Forestry, Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea (Grant No. 609001-051SB210).

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Molecular Characterization of 170 New gDNA-SSR Markers for Genetic Diversity in Button Mushroom (Agaricus bisporus)

  • An, Hyejin;Jo, Ick-Hyun;Oh, Youn-Lee;Jang, Kab-Yeul;Kong, Won-Sik;Sung, Jwa-Kyung;So, Yoon-Sup;Chung, Jong-Wook
    • Mycobiology
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    • v.47 no.4
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    • pp.527-532
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    • 2019
  • We designed 170 new simple sequence repeat (SSR) markers based on the whole-genome sequence data of button mushroom (Agaricus bisporus), and selected 121 polymorphic markers. A total of 121 polymorphic markers, the average major allele frequency (MAF) and the average number of alleles (NA) were 0.50 and 5.47, respectively. The average number of genotypes (NG), observed heterozygosity (HO), expected heterozygosity (HE), and polymorphic information content (PIC) were 6.177, 0.227, 0.619, and 0.569, respectively. Pearson's correlation coefficient showed that MAF was negatively correlated with NG (-0.683), NA (-0.600), HO (-0.584), and PIC (-0.941). NG, NA, HO, and PIC were positively correlated with other polymorphic parameters except for MAF. UPGMA clustering showed that 26 A. bisporus accessions were classified into 3 groups, and each accession was differentiated. The 121 SSR markers should facilitate the use of molecular markers in button mushroom breeding and genetic studies.

Genetic Variation in Sprout-related Traits and Microsatellite DNA Loci of Soybean

  • Lee, Suk-Ha;Kyujung Van;Kim, Moon-Young;Gwag, Jae-Gyun;Bae, Kyung-Geun;Oh, Young-Jin;Kim, Kyong-Ho;Park, Ho-Ki
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.5
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    • pp.413-418
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    • 2003
  • Genetic diversity and soybean sprout-related traits were evaluated in a total of 72 soybean accessions (60 Glycine max, 7 Glycine soja, and 5 Glycine gracilis). 100-seed weight (SW) was greatly varied and ranged from 3.2g to 32.3g in 72 soybean accessions. Positive correlation was observed between GR and hypocotyl length (HL), whereas negative correlation was observed between SW and hypocotyl diameter (HD). Re-evaluation by discarding two soybean genotypes characterized with low GR indicated that much higher correlation of sprout yield (SY) with HD and SW. Based on the principal component analysis (PCA) for sprout-related traits, 57 accessions were classified. Soybean genotypes with better traits for sprout, such as small size of seeds and high SY, were characterized with high PCA 1 and PCA 2 values. The seed size in second is small but showed low GR and SY, whereas the third has large seed, high GR and more than 400% SY. In genetic similarity analysis using 60 SSR marker genotyping, 72 accessions were classified into three major and several minor groups. Nine of twelve accessions that were identified as the representatives of soybean for sprout based on PCA were in a group by the SSR marker analysis, indicating the SSR marker selection of parental genotypes for soybean sprout improvement program.

Genetic Relationships between Gardenia jasminoides var. radicans and G. jasminoides for. grandiflora Using ISSR Markers (SSR을 이용한 꽃치자와 열매치자의 유전적 관계)

  • Huh, Man-Kyu
    • Journal of Life Science
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    • v.17 no.1 s.81
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    • pp.24-30
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    • 2007
  • Inter simple sequence repeat (ISSR) markers were performed in order to analyse the genetic relation-ships of both taxa of Gardenia jasminoides var. radicans and G. jasminoides for. grandifora. Over the 88 fragments, only one locus (ISSR-11-05) was specific to G. jasminoides var. radicans and only one (ISSR-09-05) G. jasminoides for. grandiflora. Although G. jasminoides var. radicans showed low levels of alleles and Shannon's information index than G. jasminoides for. Grandiflora, however, there was not significant differences (p > 0.05). For both taxa the mean genetic diversity of natural populations was higher than that of cultivation populations. It was suggested that domestication processes via artificial selection do not have eroded the high levels of genetic diversity. ISSR markers were more effective in classifying natural populations of wild G. jasminoides in East Asia as well as cultivated G. jasminoides. The information about the phylogenetic relationship of G. jasminoides var. radicans and its closely related species is very valuable of the systematics of genus Gardenia, the origin of cultivated G. jasminoides, and future G. jasminoides breeding.

Identification of Quantitative Trait Loci Associated with Seed Size and Weight in Soybean

  • Kim, Hong-Sik;Lee, Suk-Ha;Park, Keum-Yong;Lee, Yeong-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.45 no.4
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    • pp.227-231
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    • 2000
  • Small seed size is one of the major traits of soybean cultivars for sprouts with regard to high sprout yield. This study was conducted to identify quantitative trait loci (QTL) for seed size and weight in a set of F 6 seeds of 89 lines derived from a cross between 'Pureunkong', a soybean cultivar developed for sprouts and 'Jinpumkong 2', a soybean cultivar with no beany taste in seed due to the lack of lipoxygenases. The genetic map of 25 linkage groups with a total of 98 markers including RFLP, RAPD, SSR and classical markers was constructed from this F/sbu 5/-derived population and was used for QTL analysis. 'Pureunkong' was significantly smaller (P<0.01) than 'Jinpumkong 2' in seed size and seed weight. Genetic variation was detected and transgressive segregation was common in the population for these traits. Seven DNA markers including opT14-1600 in LG A2, opF02-400 in LG B2, Satt100, opC09-700, opG04-730 and opQll-650 in LG C2, and opY07-1100 & 1000 in LG(unknown) were significantly associated and accounted for 4.7 to 10.9% and 5.1 to 10.1 % of the phenotypic variation in seed size and seed weight, respectively. 'Pureunkong' alleles increased seed size and seed weight at the all four significant marker loci on the LG C2. These marker loci in LG C2 were closely linked and were presumed to be a single QTL. Overall, at least three independent QTLs from 3 linkage groups (A2, B2, and C2) were putatively involved in the control of seed size and seed weight.

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Evaluation of Genetic Differentiation of Albizia lucida Populations from Eastern Region of the Indian Sub-continent by ISSR Markers

  • Aparajita, Subhashree;Rout, G.R.
    • Journal of Forest and Environmental Science
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    • v.24 no.1
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    • pp.27-34
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    • 2008
  • Level and distribution of genetic diversity in seven populations of Albizia lucida Benth. in eastern region of the Indian sub-continent were estimated using ISSR markers. Relatively higher level of genetic diversity within populations was observed in seven populations of A. lucida (mean of 0.38). From the result of AMOVA, majority of genetic diversity was allocated within populations (96.2%) resulting in a moderate degree of population differentiation. The observed distribution pattern of I-SSR variant among the populations was coincided with the typical pattern of long-lived woody tree species. Genetic relationships among the populations, reconstructed by UPGMA method, revealed two genetic groups. The population of Anugul and Bargarh turned out to be the most closely related despite a distance location between them. These formations will be of great value in the development of conservation plans for species exhibiting high levels of genetic differentiation due to fragmentation, such as indication of conservation unit size, which populations should be chosen as priority in conservation plans and which samples should be introduced in areas with a low number of individuals of A. lucida.

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GENE EXPRESSION PATTERNS INDUCED BY $TAXOL^{(R)}$ AND CYCLOSPORIN A IN ORAL SQUAMOUS CELL CARCINOMA CELL LINE USING CDNA MICROARRAY (cDNA Microarray를 이용한 구강편평세포암종 세포주에서 $Taxol^{(R)}$과 Cyclosporin A로 유도된 유전자 발현양상)

  • Kim, Yong-Kwan;Lee, Jae-Hoon;Kim, Chul-Hwan
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.3
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    • pp.202-212
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    • 2006
  • It is well-known that paclitaxel($Taxol^{(R)}$), which is extracted from the pacific and English yew, has been used as a chemotherapeutic agent for ovarian carcinoma and advanced breast carcinoma and Cyclosporin A, which is highly lipophilic cyclic peptide and isolated from a fungus, has been also used as an useful immunosuppressive drug after transplantation and is associated with cellular apoptosis. Since 1953, in which James Watson, Rosalind Franklin and Francis Crick discovered the double helical structure of DNA, a few kinds of techniques for identifying gene expression have been developed. In postgenomic period, many of researchers have used the DNA microarray which is high throughput screening technique to screen large numbers of gene expression simultaneously. In this study, we searched and screened the gene expression in the oral squamous cell carcinoma cell lines treated with $Taxol^{(R)}$, cyclosporin or cyclosporin combined with $Taxol^{(R)}$ using cDNA microarray. The results were as following; 1. It was useful that the appropriate concentration of Cyclosporin A and $Taxol^{(R)}$ used in oral squamous cell carcinoma cell line was under 1${\mu}g/ml$ and 3${\mu}g/ml$. 2. In the experimental group in which $Taxol^{(R)}$ and $Taxol^{(R)}$ + Cyclosporin A were used, the cell growth was extremely decreased. 3. In the group in which Cyclosporin A was used, the MTT assay was rarely decreased which means the activity of succinyl dehydrogenase is remained in mitochondria but in the group in which the mixture of Cyclosporin A and $Taxol^{(R)}$ were used, the MTT assay was extremely decreased. 4. In the each group in which Cyclosporin A(3 ${\mu}g/ml$) and $Taxol^{(R)}$(1 ${\mu}g/ml$) were used, the cell arrest was appeared in $G_2/M$ phase and in the group in which $Taxol^{(R)}$(3 ${\mu}g/ml$) was used, the cell arrest was appeared in both S phase and $G_2/M$ phase. 5. In the oral squamous cell carcinoma cell line treated with $Taxol^{(R)}$, several genes including ANGPTL4, RALBP1 and TXNRD1, associated with apoptosis, SUI1, MAC30, RRAGA and CTGF, related with cell growth, HUS1 and DUSP5, related with cell cycle and proliferation, ATF4 and CEBPG, associated with transcription factor, BTG1 and VEGF, associated with angiogenesis, FDPS, FCER1G, GPA33 and EPHA4 associated with signal transduction and receptor activity and AKR1C2 and UGTA10 related with carcinogenesis were detected in increased levels. The genes that showed increaced expression in the oral squamous cell carcinoma cell line treated with Cyclosporin A were CYR61, SERPINB2, SSR3 and UPA3A which are known as genes associated with cell growth, carcinogenesis, receptor activity and transcription factor. The genes expressed in the HN22 cell line treated with cyclosporin combined with $taxol^{(R)}$ were ALCAM and GTSE1 associated with cancer invasiveness and cell cycle regulation.