• Title/Summary/Keyword: Molecular Marker

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Application and utilization of marker assisted selection for biotic stress resistance in hybrid rice (Oryza sativa L.)

  • Song, Jae-Young;Ouk, Sothea;Nogoy, Franz Marielle;Nino, Marjohn C.;Kwon, Soon Wook;Ha, Woongoo;Kang, Kwon-Kyoo;Cho, Yong-Gu
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.317-331
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    • 2016
  • Development of disease resistant plant is one of the important objectives in rice breeding programs because biotic stresses can adversely affect rice growth and yield losses. This study was conducted to identify lines with multiple-resistance genes to biotic stress among 173 hybrid rice breeding lines and germplasms using DNA-based markers. Our results showed that one hybrid rice line [IR98161-2-1-1-k1-3 (IR86409-3-1-1-1-1-1/IRBB66)] possessed 5 bacterial blight resistance genes (Xa4, xa5, Xa7, Xa13 and Xa21) while two hybrid rice lines [IR98161-2-1-1-k1-2 (IR86409-3-1-1-1-1-1/IRBB66) and 7292s (IR75589-31-27-8-33S(S1)/IR102758B)] possessed 3 bacterial blight resistance genes (Xa4, Xa7 and Xa21, and Xa3, Xa4 and xa5). Molecular survey on rice blast disease revealed that most of these lines had two different resistant genes. Only 11 lines possessed Pib, Pi-5, and Pi-ta. In addition, we further surveyed the distribution of insect resistant genes, such as Bph1, Bph18(t), and Wbph. Three hybrid breeding lines [IR98161-2-1-1-k1-3 (IR86409-3-1-1-1-1-1/IRBB66), IR98161-2-1-1-k1-2 (IR86409-3-1-1-1-1-1/IRBB66), and 7292s (IR75589-31-27-8-33S(S1) /IR102758B)] contained all three resistance genes. Finally, we obtained four hybrid rice breeding lines and germplasms [IR98161-2-1-1-k1-2 (IR86409-3-1-1-1-1-1/IRBB66), Damm-Noeub Khmau, 7290s, and 7292s (IR75589-31-27-8-33S(S1)/IR102758B)] possessing six-gene combination. They are expected to provide higher level of multiple resistance to biotic stress. This study is important for genotyping hybrid rice with resistance to diverse diseases and pests. Results obtained in this study suggest that identification of pyramided resistance genes is very important for screening hybrid rice breeding lines and germplasms accurately for disease and pest resistance. We will expand their cultivation safely through bioassays against diseases, pests, and disaster in its main export countries.

A Novel SNP in the Exon 8 Region of the CLMN Gene and Its Association with Marbling Score in Hanwoo (한우 CLMN 유전자 exon 8번 영역의 신규 단일염기다형과 근내지방도의 연관성에 관한 연구)

  • Shin, Sung-Chul;Chung, Eui-Ryong
    • Journal of Life Science
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    • v.29 no.12
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    • pp.1314-1320
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    • 2019
  • This study aimed to identify single nucleotide polymorphisms in exon 8 region of the calmin (CLMN) gene and to evaluate their associations with meat yield and quality traits in Hanwoo (Korean cattle). We compared gene expression pattern of CLMN between high and low marbling score groups with extremely different intramuscular fat content of the longissimus lumborum muscles in Hanwoo. The CLMN gene was highly expressed in the high marbling score group. Total of nine SNPs were identified in the exon 8 region of CLMN gene, genotyping of the SNPs was carried out using direct-sequencing analysis in Hanwoo population (n=300) to evaluate their association with meat yield and quality traits. As a result, g.23249G>C in exon 8 was significantly associated with marbling score. Animals with the CC and GC genotypes had higher marbling score than those with the GG genotype. We constructed haplotypes of CLMN gene by linkage disequilibrium analysis and analyzed association between haplotypes and meat yield and quality traits. Haplotype of CLMN gene was associated with marbling score. As a result, animals with the CC-CC haplotype (g.23249G>C and g.23465T>C SNPs) had higher marbling score than those with CT and GT haplotypes. These findings suggest that the SNPs of bovine CLMN gene may be a useful molecular marker for selection of meat quality traits in Hanwoo.

Association of A/T Rich Microsatellites with Responses to Artificial Selection for Larval Developmental Duration in the Silkworm Bombyx mori

  • Pradeep, Appukuttan Nair Retnabhavan;Awasthi, Arvind Kumar;Urs, Raje Siddaraje
    • Molecules and Cells
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    • v.25 no.4
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    • pp.467-478
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    • 2008
  • Simple sequence repeats (SSRs) and interSSR (ISSR) marker systems were used in this study to reveal genetic changes induced by artificial selection for short/long larval duration in the tropical strain Nistari of the silkworm Bombyx mori. Artificial selection separated longer larval duration (LLD) ($29.428{\pm}0.723days$) and shorter larval duration (SLD) ($22.573{\pm}0.839days$) lines from a base, inbred population of Nistari (larval span of $23.143{\pm}0.35days$). SSR polymorphism was observed between the LLD and SLD lines at one microsatellite locus, Bmsat106 ($CA_7$) and at two loci of 1074 bp and 823 bp generated with the ISSR primer UBC873. Each of these loci was present only in the LLD line. The loci segregated in the third generation of selection and were fixed in opposite directions. In the $F_2$ generation of the $LLD{\times}SLD$ lines, the alleles of Bmsat106 and $UBC873_{1074bp}$ segregated in a 1:1 ratio and the loci were present only in the LLD individuals. $UBC873_{823bp}$ was homozygous. Single factor ANOVA showed a significant association between the segregating loci and longer larval duration. Together, the two alleles contributed to an 18% increase in larval duration. The nucleotide sequences of the $UBC873_{1074bp}$ and $UBC873_{823bp}$ loci had 67% A/T content and consisted of direct, reverse, complementary and palindromic repeats. The repeats appeared to be "nested" (59%) in larger repeats or as clustered elements adjacent to other repeats. Of 203 microsatellites identified, dinucleotides (67.8%) predominated and were rich in A/T and T/A motifs. The sequences of the $UBC873_{1074bp}$ and $UBC873_{823bp}$ loci showed similarity (E = 0.0) to contigs located in Scaffold 010774 and Scaffold 000139, respectively, of the B. mori genome. BLASTN analysis of the $UBC873_{1074bp}$ sequence showed significant homology of (nt.) 45-122 with upstream region of three exons from Bombyx. The complete sequence of this locus showed ~49% nucleotide conservation with transposon 412 of Drosophila melanogaster and the Ikirara insertions of Anopheles gambiae. The A + T richness and lack of coding potential of these small loci, and their absence in the SLD line, reflect the active process of genetic change associated with the switch to short larval duration as an adaptation to the tropics.

Highly Expressed Integrin-α8 Induces Epithelial to Mesenchymal Transition-Like Features in Multiple Myeloma with Early Relapse

  • Ryu, Jiyeon;Koh, Youngil;Park, Hyejoo;Kim, Dae Yoon;Kim, Dong Chan;Byun, Ja Min;Lee, Hyun Jung;Yoon, Sung-Soo
    • Molecules and Cells
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    • v.39 no.12
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    • pp.898-908
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    • 2016
  • Despite recent groundbreaking advances in multiple myeloma (MM) treatment, most MM patients ultimately experience relapse, and the relapse biology is not entirely understood. To define altered gene expression in MM relapse, gene expression profiles were examined and compared among 16 MM patients grouped by 12 months progression-free survival (PFS) after autologous stem cell transplantation. To maximize the difference between prognostic groups, patients at each end of the PFS spectrum (the four with the shortest PFS and four with the longest PFS) were chosen for additional analyses. We discovered that integrin-${\alpha}8$ (ITGA8) is highly expressed in MM patients with early relapse. The integrin family is well known to be involved in MM progression; however, the role of integrin-${\alpha}8$ is largely unknown. We functionally overexpressed integrin-${\alpha}8$ in MM cell lines, and surprisingly, stemness features including $HIF1{\alpha}$, VEGF, OCT4, and Nanog, as well as epithelial mesenchymal transition (EMT)-related phenotypes, including N-cadherin, Slug, Snail and CXCR4, were induced. These, consequently, enhanced migration and invasion abilities, which are crucial to MM pathogenesis. Moreover, the gain of integrin-${\alpha}8$ expression mediated drug resistance against melphalan and bortezomib, which are the main therapeutic agents in MM. The cBioPortal genomic database revealed that ITGA8 have significant tendency to co-occur with PDGFRA and PDGFRB and their mRNA expression were up-regulated in ITGA8 overexpressed MM cells. In summary, integrin-${\alpha}8$, which was up-regulated in MM of early relapse, mediates EMT-like phenotype, enhancing migration and invasion; therefore, it could serve as a potential marker of MM relapse and be a new therapeutic target.

Molecular Identification of Pooideae, Poaceae in Korea (국내 농경지에 발생하는 포아풀아과 잡초의 분자생물학적 동정)

  • Lee, Jeongran;Kim, Chang-Seok;Lee, In-Yong
    • Weed & Turfgrass Science
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    • v.4 no.1
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    • pp.18-25
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    • 2015
  • A universal DNA barcoding for agricultural noxious weeds is a powerful technique for species identification without morphological knowledge, by using short sections of DNA from a specific region of the genome. Two standard barcode markers, chloroplast rbcL and matK, and a supplementary nuclear ribosomal Internal Transcribed Spacer (ITS) region were used to examine the effectiveness of the markers for Pooideae barcoding using 163 individuals of 29 taxa across 16 genera of Korean Pooideae. The rbcL and ITS revealed a good level of amplification and sequencing success while matK did not. Barcode gaps were 78.6% for rbcL, 96.2% for matK, and 91.7% for ITS, respectively. Resolving powers were 89.3% for rbcL, 92.3% for matK, and 79.1% for ITS. The matK obtained the best both barcode gap and resolving power. However, it should be considered not to employ matK for Pooideae barcode because of low rate of PCR amplification and sequencing success. As a single DNA marker, rbcL and ITS were reasonable for Pooideae barcode. Barcode gap and resolving power were increased when ITS was incorporated into the rbcL. The barcode sequences were deposited to the National Center for Biotechnology Information (NCBI) database for public use.

Usefulness of FDG-PET/CT as a Diagnostic Tool for Routine Post Therapy Evaluation in Endometrial Cancer (자궁내막암의 치료 후 루틴 추적검사 방법으로서 FDG-PET/CT의 유용성)

  • Lee, Shin-Jae;Jeon, Tae-Joo;Kim, Seung-Jo;Kim, Hee-Jin;An, Hee-Jung
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.4
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    • pp.301-308
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    • 2009
  • Purpose: The aim of this study was to evaluate the usefulness of FDG-PET/CT as follow up imaging tool in patients with endometrial cancer after therapy. Material and Methods: One hundred one patients with endometrial cancer who underwent FDG PET/CT after the treatment of this disease were included in this study population (25-79 yr old, Mean age 50.6 yr old) and all these patients also performed various laboratory and imaging studies such as serum tumor marker, CT or MRI. The lesions having increased focal FDG uptake were classified into benign, equivocal, and malignant one according to their pattern and activity. Tumor recurrence was confirmed by histopathological results and other clinical and imaging data. Results: Among the 19 patients with 30 malignant or equivocal hot uptakes, 11 of 14 patients supposed to be malignant finding in PET/CT were proved to be tumor recurrence, while one of 5 patients with equivocal lesions were recurred malignancy, Two false negative cases were turned out to be peritoneal carcinomatosis, Estimated sensitivity, specificity and accuracy of PET/CT for diagnosis of recurrence in endometrial carcinoma after treatment were 86 %, 92 % and 91 %, respectively. Positive and negative predictive values in the same issue were 63% and 98%, respectively. Conclusion: FDG-PET/CT is useful for regular work up of endometrial carcinoma after the treatment because of its high negative predictive value as well as high sensitivity and specificity.

Quantification of urea in serum by isotope dilution HPLC/MS (동위원소 희석 HPLC/MS에 의한 혈청 내 urea의 정량)

  • Lee, Hwashim;Park, Sangryoul
    • Analytical Science and Technology
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    • v.18 no.4
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    • pp.271-277
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    • 2005
  • Urea in blood has been measured as an effective marker for diagnosis of renal function. Urea which is e end-product of nitrogen containing metabolites such as proteins is filtered through glomeruli of kidneys and then excreted as urine. If the renal function is deteriorated, the urea concentration in blood will be increased, from which the healthiness of renal function is judged. In order to improve the confidence of diagnosis results, the results must keep traceability chain to certified reference materials, which was certified by primary reference method. In this study, we proposed isotope dilution-liquid chromatography/mass spectrometry (ID-LC/MS) as a candidate primary method, in which $15^N_2$-urea is used as an internal reference material. The developed method is highly accurate in principle and is convenient as it does not require cumbersome derivatization. 0.1 mmol/L ammonium chloride was selected as a mobile phase for HPLC because it provided low interference in MS analysis of relatively low molecular weighted urea. HPLC and MS were connected with an electrospray ionization (ESI) interface of positive mode, which provided high sensitivity and reproducibility. The developed method was validated with internationally recognized reference materials, and we have obtained satisfactory results in an international ring trial. The expanded uncertainty calculated according to ISO guide was 1.8% at 95% confidence interval. The developed method is being used as a primary reference measurement method such as for certification of serum certified reference materials (CRMs).

Association between the Polymorphism of the Fatty acid binding protein 5 (FABP5) Gene within the BTA 14 QTL Region and Carcass/Meat Quality Traits in Hanwoo (한우 14번 염색체 QTL 영역내 Fatty acid binding protein 5 유전자의 다형성과 도체 및 육질 형질과의 관련성 분석)

  • Heo, Kang-Nyeong;Kim, Nam-Kuk;Lee, Seung-Hwan;Kim, Nam-Young;Jeon, Jin-Tae;Park, Eung-Woo;Oh, Sung-Jong;Kim, Tae-Hun;Seong, Hwan-Hoo;Yoon, Du-Hak
    • Journal of Animal Science and Technology
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    • v.53 no.4
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    • pp.311-317
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    • 2011
  • The aim of this study was to evaluate the association between economic traits of Korean cattle (Hanwoo) and genetic variation in fatty acid binding protein 5 (FABP5) gene within QTL region of carcass weight and marbling score traits on BTA 14. We sequenced for detection of single nucleotide polymorphism (SNP) with 24 unrelated Hanwoo samples and identified four SNPs (-1141A>G, 949A>G, 969A>G and 1085C>G). Relationship between the genotypes of 583 Hanwoo individuals by PCR-RFLP and economic traits were analyzed by the mixed regression model implemented in the ASReml program. As the result of statistical analysis, SNP1 (-1141A>G) showed significant effect (p<0.003) on marbling score (MS) and SNP2 (949A>G) showed significant effect (p<0.034) on eye muscle area (EMA). Further studies are required to validate the significant SNPs in a bigger population, but the SNPs (-1141A>G and 949A>G) of FABP5 could be a genetic marker to estimate molecular breeding value (MEBV) for carcass traits in Hanwoo.

Genotypes of Lysophospholipase I (LYPLA1) Gene Associate with Carcass Traits of the Jeju Crossbred Cattle (Hanwoo × Jeju Black cattle) and Hanwoo Populations (제주도산 한우와 제주흑한우 집단의 도체형질에 대한 Lysophospholipase I (LYPLA1) 유전자형의 효과)

  • Han, Sang-Hyun;Seong, Pil-Nam;Cho, In-Cheol
    • Journal of Life Science
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    • v.27 no.7
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    • pp.760-766
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    • 2017
  • Genotypes of the nucleotide substitution g.23655332G>C of SNP marker rs385360448 at Lysophospholipase I (LYPLA1) gene intron 7, were tested for their effect on the carcass traits of Hanwoo and Jeju Crossbred cattle ($F_1$ progeny produced between Hanwoo ${\times}$ Jeju Black cattle) populations on Jeju Island. In the Hanwoo steer population, the meats containing LYPLA1 rs385360448 G/- genotypes showed significantly higher marbling scores and greater texture indices, compared to those of rs385360448 C/C homozygous animals (p<0.05). However, the LYPLA1 genotypes were not associated with the levels of carcass weight, backfat thickness, eye muscle area (EMA), meat color, and fat color (p>0.05). On the other hand, in the JCC steer population, the LYPLA1 G/- harboring meats showed significantly greater EMA levels, compared to those of C/C homozygotes (p<0.05). The results of the present study indicate that the LYPLA1 genotypes could alter the levels of intramuscular fat deposition, texture index, and eye muscle area via phospholipid metabolism in the Longissimus dorsi muscle of the cattle. These findings suggested that LYPLA1 genotypes may effect molecular genetic markers in the improvement of carcass traits of Hanwoo and Jeju Black industrial cattle populations on Jeju Island.

Mitochondrial Genetic Diversity and Phylogenetic Relationships of Siberian Flying Squirrel(Pteromys volans) Populations

  • Lee, Mu-Yeong;Park, Sun-Kyung;Hong, Yoon-Jee;Kim, Young-Jun;Voloshina, Inna;Myslenkov, Alexander;Saveljev, Alexander P.;Choi, Tae-Young;Piao, Ren-Zhu;An, Jung-Hwa;Lee, Mun-Han;Lee, Hang;Min, Mi-Sook
    • Animal cells and systems
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    • v.12 no.4
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    • pp.269-277
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    • 2008
  • Siberian flying squirrel, an endangered species in South Korea, is distributed through major mountain regions of South Korea. The number of Siberian flying squirrel(Pteromys volans) in South Korea has decreased and their habitats are fragmented and isolated because of anthropogenic activities. So far no molecular genetic data has, however, been available for their conservation and management. To obtain better information concerning genetic diversity and phylogenetic relationships of the Siberian flying squirrel in South Korea, we examined 14 individuals from South Korea, 7 individuals from Russia, and 5 individuals from northeastern China along with previously published 29 haplotypes for 1,140 bp of the mtDNA cytochrome b gene. The 14 new individuals from South Korea had 7 haplotypes which were not observed in the regions of Russia and Hokkaido. The level of genetic diversity(0.616%) in the South Korean population was lower than that in eastern Russia(0.950%). The geographical distribution of mtDNA haplotypes and reduced median network confirmed that there are three major lineages of Siberian flying squirrel, occupying; Far Eastern, northern Eurasia, and the island of Hokkaido. The South Korean population only slightly distinct from the Eurasia, and eastern Russian population, and is part of the lineage Far Eastern. Based on these, we suggest that the South Korean population could be considered to belong to one partial ESU(Far Eastern) of three partial ESUs but a different management unit. However, the conservation priorities should be reconfirmed by nuclear genetic marker and ecological data.