• 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.

• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.67-75
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• 2011

The faulty regulation of imprinting gene lead to the abnormal development of reconstructed embryo after nuclear transfer. However, the correlation between the imprinting status of donor cell and preimplantation stage of embryo development is not yet clear. In this study, to determine this correlation, we used the porcine spermatogonial stem cell (pSSC) and fetal fibroblast (pFF) as donor cells. As the results, the isolated cells with laminin matrix selection strongly expressed the GFR ${\alpha}$-1 and PLZF genes of SSCs specific markers. The pSSCs were maintained to 12 passages and positive for the pluripotent marker including OCT4, SSEA1 and NANOG. The methylation analysis of H19 DMR of pSSCs revealed that the zinc finger protein binding sites CTCF3 of H19 DMRs displayed an androgenic imprinting pattern (92.7%). Also, to investigate the reprogramming potential of pSSCs as donor cell, we compared the development rate and methylation status of H19 gene between the reconstructed embryos from pFF and pSSC. This result showed no significant differences of the development rate between the pFFs ($11.2{\pm}0.8%$) and SSCs ($13.3{\pm}1.1%$). However, interestingly, while the CTCF3 methylation status of pFF-NT blastocyst was decreased (36.3%), and the CTCF3 methylation status of pSSC-NT blastocyst was maintained. Therefore, this result suggested that the genomic imprinting status of pSSCs is more effective than that of normal somatic cells for the normal development because the maintenance of imprinting pattern is very important in early embryo stage.

• The Plant Pathology Journal
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• v.25 no.4
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• pp.408-416
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• 2009

A molecular survey was conducted to identify the presence of the bacterial blight resistance genes (Xa1, Xa4, xa5, xa13 and Xa21) in 86 accessions of aromatic rice obtained from germplasm. The results revealed that the resistance gene Xa4 (32.5%), Xa21 (17%), and xa5 (16%) were widely observed in tested rice germplasm. Among tested rice germplasm, 49 accessions showed the presence of more than one of five R genes, and 37 accessions possessed none of the R gene. TALLi and 05-IRRi-M-46 showed the presence of Xa4, xa5, xa13 and Xa21. Rice race $415{\times}Ir352$ exhibited positive amplicon for the Xa1, Xa4, xa5 and Xa21. Hyangmibyeo1hos, Ir841-85-1-1-2 and Jasmine85 showed the positive amplicon for the Xa1, Xa4 and xa5 genes. Yekywin Yinkya Hmwe and Khao Dawk Mali105 showed the presence of Xa1, Xa4 and Xa21 gene. Masino Basmati showed the presence of xa5, xa13, Xa21 genes. Xa1 and Xa21 genes were noticed in Mihayngbyeo, Tarana Deshi, Mayataung and AZUCENA. Hyangmibyeo2ho, Basmati 6311 and Basmati405 possessed only two R genes such as Xa4 and xa5, and xa5 and xa13, respectively. The evaluation results of bacterial blight resistance genes in aromatic rice germplasm will help in breeding of multi disease resistant varieties.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.10
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• pp.1490-1495
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• 2007

Insulin-like growth factor binding protein-4 (IGFBP-4) is a member of the IGF super family, and regulates the action of IGFs. The polymorphism of porcine IGFBP-4 gene in 17 pig breeds (total n = 570) was detected by PCR-SSCP, and alleles A and B were detected. In these pig breeds, it was found that exotic pig breeds carried high frequencies of allele A, while Chinese native pig breeds carried high frequencies of allele B. The role of porcine IGFBP-4 was investigated in 172 F2 offspring of a $Lantang{\times}Lantang $ population. Forty eight growth traits were recorded for analyzing the association between IGFBP-4 gene polymorphism and quantitative performance traits. In this resource family, pigs with AA genotype had higher fore-body weight, bone weight of mid-body, bone weight of rear-body, fore-leg weight and rear-leg weight than those pigs with BB genotype (p<0.05); while pigs which carried BB genotype had higher back-fat thickness at C point and lard weight than those pigs with AA genotype (p<0.05); pigs with AA genotype had higher body weight than those with BB genotype; for meat quality traits, pigs with AA genotype had higher meat color than those of BB genotype (p<0.01), and pigs with BB genotype had higher marbling than those of AA and AB genotypes (p<0.01 and p<0.05, respectively).Based on these results, it is necessary to do more studies on IGFBP-4 before using the IGFBP-4 locus for the application of marker-assisted selection programs.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.10
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• pp.1375-1378
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• 2005

The objective of this study was to investigate polymorphisms of BMPRIB (bone morphogenetic protein type IB receptor) gene and its effect on litter size traits in sheep. Three populations including 101 Small Tailed Han sheep, 79 Poll Dorset and 81 hybrids (Poll Dorset${\times}$Small Tailed Han sheep) were used to detect the polymorphisms in exon 6 region of sheep BMPRIB gene. A fragment of approximately 190bp was amplified by one pair of primers, the polymorphism was revealed from the analysis of three populations by the technique of PCR -SSCP, and a mutation from A to G at 746 of the coding region was confirmed by sequencing in several individual. Statistical results indicated the distribution of allele B (with a A$\longrightarrow$G mutation) and A (without mutation) or genotype AA, AB and BB frequencies differed in three populations. BB genotype (44.55%) and B allele (66.34%) frequencies of Small Tailed Han sheep were higher than those of the others. Analysis of variance showed that the polymorphism of BMPRIB gene was associated with positive effect on litter size traits. The means of genotype BB and AB were about 1.04 and 0.74 more than genotype AA for litter size (p<0.05). Analysis of BMPRIB genotype effects on litter size in three populations indicates the existence of genotype BB or B allele increases the litter size. It suggested that the polymorphism in exon 6 (at 746 in the coding region) of sheep BMPRIB gene may be used as a marker for early selection of prolificacy in sheep.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.5
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• pp.669-676
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• 2007

This study was conducted to detect quantitative trait loci (QTL) affecting growth and body composition in an $F_2$ reference population of Korean native pig and Landrace crossbreds. The three-generation mapping population was generated with 411 progeny from 38 $F_2$ full-sib families, and 133 genetic markers were used to produce a sex-average map of the 18 autosomes. The data set was analyzed using least squares Mendelian and parent-of-origin interval-mapping models. Lack-of-fit tests between the models were used to characterize QTL for mode of expressions. A total of 8 (39) QTL were detected at the 5% genome (chromosome)-wise level for the 17 analyzed traits. Of the 47 QTL detected, 21 QTL were classified as Mendelian expressed, 13 QTL as paternally expressed, 6 QTL as maternally expressed, and 7 QTL as partially expressed. Of the detected QTL at 5% genome-wise level, two QTL had Mendelian mode of inheritance on SSC6 and SSC9 for backfat thickness and bone weight, respectively, two QTL were maternally expressed for leather weight and front leg weight on SSC6 and SSC12, respectively, one QTL was paternally expressed for birth weight on SSC4, and three QTL were partially expressed for hot carcass weight and rear leg weight on SSC6, and bone weight on SSC13. Many of the Mendelian QTL had a dominant (complete or overdominant) mode of gene action, and only a few of the QTL were primarily additive, which reflects that heterosis for growth is appreciable in a cross between Korean native pig and Landrace. Our results indicate that alternate breed alleles of growth and body composition QTL are segregating between the two breeds, which could be utilized for genetic improvement of growth via marker-assisted selection.

• Journal of Microbiology and Biotechnology
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• v.25 no.2
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• pp.196-205
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• 2015

A Sulfolobus-E. coli shuttle vector for an efficient expression of the target gene in S. acidocaldarius strain was constructed. The plasmid-based vector pSM21 and its derivative pSM21N were generated based on the pUC18 and Sulfolobus cryptic plasmid pRN1. They carried the S. solfataricus P2 pyrEF gene for the selection marker, a multiple cloning site (MCS) with C-terminal histidine tag, and a constitutive promoter of the S. acidocaldarius gdhA gene for strong expression of the target gene, as well as the pBR322 origin and ampicillin-resistant gene for E. coli propagation. The advantage of pSM21 over other Sulfolobus shuttle vectors is that it contains a MCS and a histidine tag for the simple and easy cloning of a target gene as well as one-step purification by histidine affinity chromatography. For successful expression of the foreign genes, two genes from archaeal origins (PH0193 and Ta0298) were cloned into pSM21N and the functional expression was examined by enzyme activity assay. The recombinant PH0193 was successfully expressed under the control of the gdhA promoter and purified from the cultures by His-tag affinity chromatography. The yield was approximately 1 mg of protein per liter of cultures. The enzyme activity measurements of PH0913 and Ta0298 revealed that both proteins were expressed as an active form in S. acidocaldarius. These results indicate that the pSM21N shuttle vector can be used for the functional expression of foreign archaeal genes that form insoluble aggregates in the E. coli system.

• Korean Journal of Plant Resources
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• v.29 no.6
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• pp.658-669
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• 2016

Rice blast, caused by a fungus Magnaporthe oryzae, is one of the most devastating diseases of rice worldwide. Analyzing the valuable genetic resources is important in making progress towards blast resistance. Molecular screening of major rice blast resistance (R) genes was determined in 2,509 accessions of rice germplasm from different geographic regions of Asia and Europe using PCR based markers which showed linkage to twelve major blast R genes, Pik-p, Pi39, Pit, Pik-m, Pi-d(t)2, Pii, Pib, Pik, Pita, Pita/Pita-2, Pi5, and Piz-t. Out of 2,509 accessions, only two accessions had maximum nine blast resistance genes followed by eighteen accessions each with eight R genes. The polygenic combination of three genes was possessed by maximum number of accessions (824), while among others 48 accessions possessed seven genes, 119 accessions had six genes, 267 accessions had five genes, 487 accessions had four genes, 646 accessions had two genes, and 98 accessions had single R gene. The Pik-p gene appeared to be omnipresent and was detected in all germplasm. Furthermore, principal component analysis (PCA) indicated that Pita, Pita/Pita-2, Pi-d(t)2, Pib and Pit were the major genes responsible for resistance in the germplasm. The present investigation revealed that a set of 68 elite germplasm accessions would have a competitive edge over the current resistance donors being utilized in the breeding programs. Overall, these results might be useful to identify and incorporate the resistance genes from germplasm into elite cultivars through marker assisted selection in rice breeding.

• Korean Journal of Medicinal Crop Science
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• v.24 no.1
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• pp.55-61
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• 2016

Background : Plant breeding requires the collection of genetically diverse genetic resources. Studies on the characteristics of Platycodon grandiflorum resources have not been carried out so far. The present study was carried out to discriminate P. grandiflorum based on morphological characteristics and genetic diversity using simple sequence repeat (SSR) markers. Methods and Results :We collected 11 P. grandiflorum cultivars: Maries II, Hakone double white, Hakone double blue, Fuji white, Fuji pink, Fuji blue, Astra white, Astra pink, Astra blue, Astra semi-double blue and Jangbaek. Analyses of the morphological characteristics of the collection were conducted for aerial parts (flower, stem and leaf) and underground parts (root). Next, the genetic diversity of all P. grandiflorum resources was analyzed using SSR markers employing the DNA fragment analysis method. We determined that the 11 P. grandiflorum cultivars analyzed could be classified by plant length, leaf number and root characteristic. Based on the genetic diversity analysis, these cultivars were classified into four distinct groups. Conclusions : These findings could be used for further research on cultivar development using molecular breeding techniques and for conservation of the genetic diversity of P. grandiflorum. Moreover, the markers could be used for genetic mapping of the plant and marker-assisted selection for crop breeding.

• Korean Journal of Microbiology
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• v.41 no.2
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• pp.146-151
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• 2005

To construct an amylolytic industrial strain of Saccharomyces cerevisiae, the glucoamylase cDNA gene (GAl) from Aspergillus awamori was expressed under the control of the alcohol dehydrogenase gene promoter (ADC1p) and integrated into the chromosomes of industrial S. cerevisiae. An integrative cassette lacking bacterial ampicillin resistance gene but containing the GA1 gene, $\delta$ sequences of Ty1 retrotransposon as target sites for homologous recombination and S. cerevisiae aureobasidin A resistance gene (AUR1-C) as the selection marker was constructed to obtain a strain eligible for commercial use. Industrial S. cerevisiae transformed with this 15-integrative cassette efficiently secreted glucoamylase into the medium and grew on starch as the sole carbon source. The transformants were mitotically stable for 100 generations in nonselective medium.