• Title/Summary/Keyword: genetic markers

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Discordance between Morphological and Molecular Variations of the Genus Macroramphosus (Macroramphosidae) from Korea (한국산 대주둥치속(대주둥치과) 어류의 형태와 분자 변이의 불일치)

  • Sohn, Min-Soo;Kim, Jin-Koo
    • Korean Journal of Ichthyology
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    • v.32 no.4
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    • pp.199-209
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    • 2020
  • In order to clarify the taxonomic status of the Korean Macroramphosus species, which were previously confused, we investigated morphological and molecular variations of Macroramphosus (18 individuals) from Korea, and Macroramphosus (35 individuals) from Japan and Taiwan, and compared with those of M. scolopax from type locality (Mediterranean Sea). Although the Korean and Japanese specimens of Macroramphosus were clearly divided into two types in the first dorsal spine length (22.8~32.1% in A-type vs. 15.6~21.4% in B-type), distance between the first dorsal fin and second dorsal fin (6.4~9.7% vs. 8.6~13.3%), and body depth (20.0~28.0% vs. 17.3~22.6%), no genetic differences among all individuals of longspine snipefish between them were found at the specific level [d=0.0~3.3% in control region (CR); 0.0~1.3% in cytochrome b (cytb); 0.0~0.5% in cytochrome c oxidase subunit I (COI)]. Whereas, they were well distinguished in genetics (9.9~11.5% in CR; 3.8~4.6% in cytb; 1.2~3.6% in COI) from those of M. scolopax in Mediterranean Sea. It needs the scientific name of the longspine snipefish (M. scolopax) in Korea be changed as M. japonicus (and/or M. sagifue). However, our results could not find evidence of consistency between morphological and mitochondrial DNA variations which suggests that their differentiation event may occur fairly recently. Further studies using more sensitive markers such as microsatellite are needed to clarify the degree of gene flow between the two types.

Mutations in GJB2 as Major Causes of Autosomal Recessive Non-Syndromic Hearing Loss: First Report of c.299-300delAT Mutation in Kurdish Population of Iran

  • Azadegan-Dehkordi, Fatemeh;Bahrami, Tayyebe;Shirzad, Maryam;Karbasi, Gelareh;Yazdanpanahi, Nasrin;Farrokhi, Effat;Koohiyan, Mahbobeh;Tabatabaiefar, Mohammad Amin;Hashemzadeh-Chaleshtori, Morteza
    • Journal of Audiology & Otology
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    • v.23 no.1
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    • pp.20-26
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    • 2019
  • Background and Objectives: Autosomal recessive non-syndromic hearing loss (ARNSHL) with genetic origin is common (1/2000 births). ARNSHL can be associated with mutations in gap junction protein beta 2 (GJB2). To this end, this cohort investigation aimed to find the contribution of GJB2 gene mutations with the genotype-phenotype correlations in 45 ARNSHL cases in the Kurdish population. Subjects and Methods: Genomic DNA was extracted from a total of 45 ARNSHL families. The linkage analysis with 3 short tandem repeat markers linked to GJB2 was performed on 45 ARNSHL families. Only 9 of these families were linked to the DFNB1 locus. All the 45 families who took part were sequenced for confirmation linkage analysis (to perform a large project). Results: A total of three different mutations were determined. Two of which [c.35delG and c.-23+1G>A (IVS1+1G>A)] were previously reported but (c.299-300delAT) mutation was novel in the Kurdish population. The homozygous pathogenic mutations of GJB2 gene was observed in nine out of the 45 families (20%), also heterozygous genotype (c.35delG/N)+(c.-23+1G>A/c.-23+1G>A) were observed in 4/45 families (8.8%). The degree of hearing loss (HL) in patients with other mutations was less severe than patients with c.35delG homozygous mutation (p<0.001). Conclusions: Our data suggest that GJB2 mutations constitute 20% of the etiology of ARNSHL in Iran; moreover, the c.35delG mutation is the most common HL cause in the Kurdish population. Therefore, these mutations should be included in the molecular testing of HL in this population.

Mutations in GJB2 as Major Causes of Autosomal Recessive Non-Syndromic Hearing Loss: First Report of c.299-300delAT Mutation in Kurdish Population of Iran

  • Azadegan-Dehkordi, Fatemeh;Bahrami, Tayyebe;Shirzad, Maryam;Karbasi, Gelareh;Yazdanpanahi, Nasrin;Farrokhi, Effat;Koohiyan, Mahbobeh;Tabatabaiefar, Mohammad Amin;Hashemzadeh-Chaleshtori, Morteza
    • Korean Journal of Audiology
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    • v.23 no.1
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    • pp.20-26
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    • 2019
  • Background and Objectives: Autosomal recessive non-syndromic hearing loss (ARNSHL) with genetic origin is common (1/2000 births). ARNSHL can be associated with mutations in gap junction protein beta 2 (GJB2). To this end, this cohort investigation aimed to find the contribution of GJB2 gene mutations with the genotype-phenotype correlations in 45 ARNSHL cases in the Kurdish population. Subjects and Methods: Genomic DNA was extracted from a total of 45 ARNSHL families. The linkage analysis with 3 short tandem repeat markers linked to GJB2 was performed on 45 ARNSHL families. Only 9 of these families were linked to the DFNB1 locus. All the 45 families who took part were sequenced for confirmation linkage analysis (to perform a large project). Results: A total of three different mutations were determined. Two of which [c.35delG and c.-23+1G>A (IVS1+1G>A)] were previously reported but (c.299-300delAT) mutation was novel in the Kurdish population. The homozygous pathogenic mutations of GJB2 gene was observed in nine out of the 45 families (20%), also heterozygous genotype (c.35delG/N)+(c.-23+1G>A/c.-23+1G>A) were observed in 4/45 families (8.8%). The degree of hearing loss (HL) in patients with other mutations was less severe than patients with c.35delG homozygous mutation (p<0.001). Conclusions: Our data suggest that GJB2 mutations constitute 20% of the etiology of ARNSHL in Iran; moreover, the c.35delG mutation is the most common HL cause in the Kurdish population. Therefore, these mutations should be included in the molecular testing of HL in this population.

Species Identification and Monitoring of Labeling Compliance for Commercial Pufferfish Products Sold in Korean On-line Markets (국내 온라인 유통 복어 제품의 종판별 및 표시사항 모니터링 연구)

  • Ji Young Lee;Kun Hee Kim;Tae Sun Kang
    • Journal of Food Hygiene and Safety
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    • v.38 no.6
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    • pp.464-475
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    • 2023
  • In this study, based on an analysis of two DNA barcode markers (cytochrome c oxidase subunit I and cytochrome b genes), we performed species identification and monitored labeling compliance for 50 commercial pufferfish products sold in on-line markets in Korea. Using these barcode sequences as a query for species identification and phylogenetic analysis, we screened the GenBank database. A total of seven pufferfish species (Takifugu chinensis, T. pseudommus, T. xanthopterus, T. alboplumbeus, T. porphyreus, T. vermicularis, and Lagocephalus cheesemanii) were identified and we detected 35 products (70%) that were non-compliant with the corresponding label information. Moreover, the labels on 12 commercial products contained only the general common name (i.e., pufferfish), although not the scientific or Korean names for the 21 edible pufferfish species. Furthermore, the proportion of mislabeled highly processed products (n = 9, 81.8%) was higher than that of simply processed products (n = 26, 66.7%). With respect to the country of origin, the percentage of mislabeled Chinese products (n = 8, 80%) was higher than that of Korean products (n = 26, 66.7%). In addition, the market and dialect names of different pufferfish species were labeled only as Jolbok or Milbok, whereas two non-edible pufferfish species (T. vermicularis and T. pseudommus) were used in six commercial pufferfish products described as JolboK and Gumbok on their labels, which could be attributable to the complex classification system used for pufferfish. These monitoring results highlight the necessity to develop genetic methods that can be used to identify the 21 edible pufferfish species, as well as the need for regulatory monitoring of commercial pufferfish products.

Studies of Molecular Breeding Technique Using Genome Information on Edible Mushrooms

  • Kong, Won-Sik;Woo, Sung-I;Jang, Kab-Yeul;Shin, Pyung-Gyun;Oh, Youn-Lee;Kim, Eun-sun;Oh, Min-Jee;Park, Young-Jin;Lee, Chang-Soo;Kim, Jong-Guk
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.53-53
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    • 2015
  • Agrobacterium tumefaciens-mediated transformation(ATMT) of Flammulina velutipes was used to produce a diverse number of transformants to discover the functions of gene that is vital for its variation color, spore pattern and cellulolytic activity. Futhermore, the transformant pool will be used as a good genetic resource for studying gene functions. Agrobacterium-mediated transformation was conducted in order to generate intentional mutants of F. velutipes strain KACC42777. Then Agrobacterium tumefaciens AGL-1 harboring pBGgHg was transformed into F. velutipes. This method is use to determine the functional gene of F. velutipes. Inverse PCR was used to insert T-DNA into the tagged chromosomal DNA segments and conducting sequence analysis of the F. velutipes. But this experiment had trouble in diverse morphological mutants because of dikaryotic nature of mushroom. It needed to make monokaryotic fruiting varients which introduced genes of compatible mating types. In this study, next generation sequencing data was generated from 28 strains of Flammulina velutipes with different phenotypes using Illumina Hiseq platform. Filtered short reads were initially aligned to the reference genome (KACC42780) to construct a SNP matrix. And then we built a phylogenetic tree based on the validated SNPs. The inferred tree represented that white- and brown- fruitbody forming strains were generally separated although three brown strains, 4103, 4028, and 4195, were grouped with white ones. This topological relationship was consistently reappeared even when we used randomly selected SNPs. Group I containing 4062, 4148, and 4195 strains and group II containing 4188, 4190, and 4194 strains formed early-divergent lineages with robust nodal supports, suggesting that they are independent groups from the members in main clades. To elucidate the distinction between white-fruitbody forming strains isolated from Korea and Japan, phylogenetic analysis was performed using their SNP data with group I members as outgroup. However, no significant genetic variation was noticed in this study. A total of 28 strains of Flammulina velutipes were analyzed to identify the genomic regions responsible for producing white-fruiting body. NGS data was yielded by using Illumina Hiseq platform. Short reads were filtered by quality score and read length were mapped on the reference genome (KACC42780). Between the white- and brown fruitbody forming strains. There is a high possibility that SNPs can be detected among the white strains as homozygous because white phenotype is recessive in F. velutipes. Thus, we constructed SNP matrix within 8 white strains. SNPs discovered between mono3 and mono19, the parental monokaryotic strains of 4210 strain (white), were excluded from the candidate. If the genotypes of SNPs detected between white and brown strains were identical with those in mono3 and mono19 strains, they were included in candidate as a priority. As a result, if more than 5 candidates SNPs were localized in single gene, we regarded as they are possibly related to the white color. In F. velutipes genome, chr01, chr04, chr07,chr11 regions were identified to be associated with white fruitbody forming. White and Brown Fruitbody strains can be used as an identification marker for F. veluipes. We can develop some molecular markers to identify colored strains and discriminate national white varieties against Japanese ones.

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SSR Profiling and Its Variation in Soybean Germplasm (콩 유전자원의 SSR Profiling과 변이)

  • Yoon, Mun-Sup;Lee, Jeong-Ran;Baek, Hyung-Jin;Cho, Gyu-Taek;Kim, Chang-Yung;Cho, Yang-Hee;Kim, Tae-San;Cho, Eun-Gi
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.1
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    • pp.81-88
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    • 2007
  • The evaluation of soybean germplasm has mainly been carried out by morphological characters at Genetic Resources Division, Rural Development Administration (RDA). However, this information has been limited serving a diverse information for user and effectively managing the soybean germplasm. To resolve this problem, soybean collection conserved at RDA gene bank was profiled using nine soybean SSR (Simple Sequence Repeat) markers. Soybean SSR allele was confirmed using genescan and genotyper softwares of automatic sequencer for accurate genotyping of each accession and continuous accumulation of data. SSR profiling of soybean germplasm has been carried out from 2,855 (Satt458) to 4,368 (Satt197) accessions by locus. The number of allele revealed 267 with an average of 29.6 in total accession, and varied from a low of 21 (Satt532 and Satt141) to a high of 58 (Sat_074). Although the number of accessions of wild soybean is less than that of soybean landraces, Korean wild soybean is more variable than other soybean landraces populations in total number of alleles. However, Korean soybean landraces were more variable than Korean wild soybeans in 5 loci. In the allele frequency, wild soybean accessions showed an even distribution in all alleles and higher distribution in low ladder than in high ladder. Also, Korean soybean landraces revealed a high condensed frequency in Satt286 (202 bp, 232 bp), Chinese soybean landraces in Satt197 (171 bp) and Satt458 (173 bp), and Japanese soybean landraces in Sat_074 (244 bp) and Satt458 (170 bp). These SSR profile information will be provided as indications of redundancies or omissions of accessions and can aid in managing soybean collection held at RDA gene bank. The information on diversity analysis could help to enlarge the genetic diversity of materials in breeding program, and could be used to develop a core collection of soybean germplasm.

Improvement of Selection Efficiency for Bacterial Blight Resistance Using SNP Marker in Rice (SNP 마커를 이용한 벼 흰잎마름병 저항성 선발 효율 증진)

  • Shin, Woon-Chul;Baek, So-Hyeon;Seo, Chun-Sun;Kang, Hyeon-Jung;Kim, Chung-Kon;Shin, Mun-Sik;Lee, Gang-Seob;Hahn, Jang-Ho;Kim, Hyun-Soon
    • Journal of Plant Biotechnology
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    • v.33 no.4
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    • pp.309-313
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    • 2006
  • Discovery of single nucleotide polymorphisms (SNPs), including small insertions and deletions, is one of the hot topics in genetic research. The most common type of sequence variant consists of single base differences or small insertions and deletions at specific nucleotide positions. Significance of SNPs in rice is increasing for genetic research, positional cloning and molecular breeding. $F_2$ 170 lines and $F_3$ 194 lines derived from Sangjuchalbyeo/HR13721-53-3-1-3-3-2-2 Were used for Searching SNP markers related to bacterial blight resistance. Sangjuchalbyeo is susceptible to bacterial blight, but HR13721-53-3-1-3-3-2-2 has Xa1 gene resistant to bacterial blight. Individual lines were inoculated with $K_1$ race of bacterial blight and resistant or susceptible was evaluated after 3 weeks from inoculation. The genotypes of population were analysed by PCR-RFLP for SNP marker developing. The segregation of $F_2\;and\;F_3$ population showed almost 3:1, 1:1 ratio, respectively. Analysis of genotype using SNP marker is capable of confirming resistance for $K_1$ race and genotype through amplifying the gene using 16PFXal primer and digested the PCR product with Eco RV. There were close relation between resistance test for $K_1$ race and SNP marker genotype. Especially, DNA analysis using SNP marker is capable of judging homozygote/heterozygote in $F_2$ population compared with resistant test for Kl race. So, it seems to improve the selection efficiency in disease resistant breeding.

Agronomic and Genetic Evaluation on a Dull Mutant Line Derived from the Sodium Azide Treated 'Namil', a Non-Glutinous Japonica Rice (남일벼 돌연변이 유래 중간찰 계통의 작물학적 특성 및 배유특성 지배유전자위 표지)

  • Chun, Jae-Buhm;Jeung, Ji-Ung;Cho, Seong-Woo;Kim, Woo-Jae;Ha, Ki-Young;Kang, Kyung-Ho;Ko, Jae-Kwon;Kim, Hyun-Soon;Kim, Bo-Kyeong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.4
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    • pp.448-457
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    • 2015
  • Developing rice lines with various amylose contents is necessary to diverse usages of rice in terms of raw materials for processed food production, and thereby to promote rice consumption in Korea. A rice mutant line, 'Namil(SA)-dull1' was established through sodium azide mutagenesis on 'Namil', a non-glutinous Korean Japonica rice cultivar. Namil(SA)-dull1' had dull endosperm characteristics and the evaluated amylose content was 12.2%. A total of 94 F2 progenies from a cross between 'Namil(SA)-dull1' and 'Milyang23', a non-glutinous Tongil-type rice cultivar, was used for genetic studies on the endosperm amylose content. Association analyses, between marker genotypes of 53 SSR anchor markers and evaluated amylose contents of each 94 F2:3 seeds, initially localized rice chromosome 6 as the harboring place for the modified allele(s) directing low amylose content of 'Namil(SA)-dull1'. By increasing SSR marker density on the putative chromosomal region followed by association analyses, the target region was narrowed down 0.94 Mbp segment, expanding from 28.95 Mbp to 29.89 Mbp, on rice chromosome 6 pseudomolecule. Among the SSR loci, RM7555 explained 84.2% of total variation of amylose contents in the $F_2$ population. Further physical mapping on the target region directing low amylose content of 'Namil(SA)-dull1' would increase the breeding efficiency in developing promising rice cultivars with various endosperm characteristics.

Single Nucleotide Polymorphisms (SNPs) Discovery in GHSR Gene and Their Association Analysis with Economic Traits in Korean Native Chickens (GHSR 유전자 내 유전변이의 탐색과 한국재래계의 성장 및 산란 특성에 미치는 연관성 분석)

  • Choi, So-Young;Hong, Min-Wook;Yang, Song-Yi;Kim, Chong-Dae;Jeong, Dong Kee;Hong, Yeong Ho;Lee, Sung-Jin
    • Korean Journal of Poultry Science
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    • v.43 no.4
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    • pp.273-279
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    • 2016
  • Recently, it was reported that certain polymorphisms in the growth hormone secretagogue receptor gene (GHSR) are associated with the growth of chickens. However, the correlation between GHSR polymorphisms and economic traits has not been investigated in Korean native chickens (KNCs). Therefore, the objective of this study was to confirm the suitability of the GHSR gene as a candidate for genomic selection and identify a genetic marker for KNCs. A total of 220 KNCs from six breeds raised at the National Institute of Animal Science were genotyped for the c.739+726 SNP in the GHSR gene using polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP), and the sequence for a subset of 30 birds was analyzed using direct sequencing. The association between the SNP genotypes and the economic traits of the KNCs was analyzed using the statistical package for the social science (SPSS) software program. The association analysis between the c.739+726T>C SNP and economic traits revealed that the SNP was significantly associated with body weight at 150 and 270 days (BW150 and BW270, respectively) in all KNCs (p<0.01), BW150 in KNC (Gary) (p<0.05), and egg production number in KNC (White, p<0.05). In addition, the SNPs discovered using direct sequencing (513A>G, 517A>T) had a significant effect on the body weight and egg production traits (p<0.05). In conclusion, these results might be useful as a basis for studies on the improvement of KNC breeds. Furthermore, these results suggest that the SNPs (c.739+726T>C, 513A>G, and 517A>T) located in the GHSR gene could be useful molecular genetic markers for KNCs.

Strain Improvement of the Genus Pleurotus by Protoplast Fusion (원형질체(原形質體) 융합(融合)에 의한 느타리버섯속(屬)의 품종개발(品種開發))

  • Yoo, Young-Bok;You, Chang-Hyun;Cha, Dong-Yeul
    • The Korean Journal of Mycology
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    • v.21 no.3
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    • pp.200-211
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    • 1993
  • Somatic hybrids of Pleurotus florida ASI 2016 and Pleurotus ostreatus ASI 2018 were obtained by protoplast fusion. The 40 fusants($P1{\sim}P40$) was examined for the yield on fermented and pasteurized rice straw in a tray. The carpophore yield of them were showed as the range of $27.0{\sim}155.2$, based on parental values of 100(ASI 2018), The pilei of fusants between orange white colored P. florida and dark grey colored P. ostreatus had mixed colors in the young stage. Other breeding programmes were performed to improve new varieties with high yield and good quality. A new oyster mushroom variety, Wonhyeongneutaribeosus(P72), was developed at the Agricultural Sciences Institute, Rural Development Administration in 1990. This P.florida-ostreatus-ostreatus hybrid P72 was selected from 38 protoplast fusion products($P41{\sim}P78$) between P.florida-ostreatus recombinant P5-M 43-arg rib and P. ostreatus ASI 2-13-0 2001-19-pro orn. The yield indexes of 38 hybrids ranged $40.5{\sim}152.7$ compared with the parental values of 100(ASI 2001). Hybrid P72 was characterized by the large fruiting bundle of semispherical shape with long stipe and by the small and circular pileus, resulting in lower harvesting cost. A significant increase in carpophore production was observed in somatic hybrids of protoplasts due to heterosis. A comparision of hybrid with parents P72 was made using isozyme analysis. The esterase banding patterns could be characterized by new bands in the hybrids. Seven fusion products of four crosses between P.florida ASI 2016 and P. ostreatus ASI 2018 were analysed with respect to the distribution of progenies and segregation of gene markers by random basidiospore analysis. Segregation of alleles should yield progeny of four genotypes in a Mendelian ratio of 1 : 1 : 1 : 1 for prototrophs, auxotrophs of one parental type, auxotrophs of the other parental type, and auxotrophic recombinants, respectively. However, five fusants of them did not detect one parental, P.ostreatus, type. Basidiospores could yield progeny of 16 genotypes in the cross of one of the recombinant P5-M43-arg $rib{\times}P. ostreatus$ ASI 2-13-pro orn but the segregants of three fusants were not detected clearly. The allele ratio of loci could be expected 1 : 1 : 1 : 1 for arg, rib, pro and orn. The ratio, however, would be changed to 4 : 1 : 1 : 1 with increasing proportion of argo In almost all the fusants, prototrophic recombinants were recovered in large numbers against auxotrophic markers. Parental genotypes were recovered with the recombinant progeny amounting to $38.68{\sim}99.56%$. The analysis provides proof of heterokaryosis and strong evidence for haploidy of vegetative nuclei, a sexual cycle consisting of nuclear fusion and meiosis.

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