• Journal of Animal Science and Technology
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• v.63 no.2
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• pp.262-271
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• 2021

Communication among epididymal epithelial cells creates the best luminal condition where spermatozoa mature, transport and are stored. Vacuolar ATPase (V-ATPase) and cytokeratin 5 (KRT5) have been used as signal indicators for clear and basal cells of the epididymal epithelium, respectively, in mice, rats, bats, and pigs; however, these two markers have not yet been described in the epididymis of bulls. Here, we examined the presence and distribution of the B1 subunit of V-ATPase (B1-VATPase) and KRT5 in the distinct regions of adult bovine epididymides, specifically, the caput, corpus, and cauda. Immunofluorescence staining and confocal microscopy showed that narrow shaped-clear cells were placed in the caput and corpus regions of the bovine epididymis; however, they were absent in the cauda epididymis. In addition, B1-VATPase was highly expressed in the cauda spermatozoa; however, it was rarely detected in the caput spermatozoa. On the other hand, KRT5-positive cells, basal cells, were maintained beneath the basal lamina and they had the traditional form with a dome-shaped morphology from the caput to cauda region of the bovine epididymis. The co-expression of B1-VATPase and KRT5 was confined to basal cells placed in the basal region of the epithelium. In summary, 1) clear cells were present with region-specific localization, 2) B1-VATPase was present in the corpus and cauda spermatozoa but absent in the caput, 3) co-expressed cells with B1-VATPase and KRT5 were present in the adult bovine epididymis, and 4) B1-VATPase was not a specific marker for clear cells in the bovine epididymis. Therefore, the perfect epididymal luminal condition created by the specific expression and localization patterns of B1-VATPase might be necessary to obtain fertilizing capacity of spermatozoa in the bovine epididymis.

• Journal of Periodontal and Implant Science
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• v.52 no.3
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• pp.242-257
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• 2022

Purpose: This study investigated periodontal ligament (PDL) restoration in osseointegrated implants using stem cells. Methods: Commercial pure titanium and zirconium oxide (zirconia) were coated with beta-tricalcium phosphate (β-TCP) using a long-pulse Nd:YAG laser (1,064 nm). Isolated bone marrow mesenchymal cells (BMMSCs) from rabbit tibia and femur, isolated PDL stem cells (PDLSCs) from the lower right incisor, and co-cultured BMMSCs and PDLSCs were tested for periostin markers using an immunofluorescent assay. Implants with 3D-engineered tissue were implanted into the lower right central incisors after extraction from rabbits. Forty implants (Ti or zirconia) were subdivided according to the duration of implantation (healing period: 45 or 90 days). Each subgroup (20 implants) was subdivided into 4 groups (without cells, PDLSC sheets, BMMSC sheets, and co-culture cell sheets). All groups underwent histological testing involving haematoxylin and eosin staining and immunohistochemistry, stereoscopic analysis to measure the PDL width, and field emission scanning electron microscopy (FESEM). The natural lower central incisors were used as controls. Results: The BMMSCs co-cultured with PDLSCs generated a well-formed PDL tissue that exhibited positive periostin expression. Histological analysis showed that the implantation of coated (Ti and zirconia) dental implants without a cell sheet resulted in a well-osseointegrated implant at both healing intervals, which was confirmed with FESEM analysis and negative periostin expression. The mesenchymal tissue structured from PDLSCs only or co-cultured (BMMSCs and PDLSCs) could form a natural periodontal tissue with no significant difference between Ti and zirconia implants, consequently forming a biohybrid dental implant. Green fluorescence for periostin was clearly detected around the biohybrid implants after 45 and 90 days. FESEM showed the invasion of PDL-like fibres perpendicular to the cementum of the bio-hybrid implants. Conclusions: β-TCP-coated (Ti and zirconia) implants generated periodontal tissue and formed biohybrid implants when mesenchymal-tissue-layered cell sheets were isolated from PDLSCs alone or co-cultured BMMSCs and PDLSCs.

• Animal Bioscience
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• v.36 no.9
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• pp.1357-1366
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• 2023

Objective: This study aimed to identify the single-nucleotide polymorphisms (SNPs) in the dual-specificity phosphatase 8 (DUSP8) and insulin-like growth factor 2 (IGF2) genes and to explore their effects on inosine-5'-monophosphate (IMP), inosine, and hypoxanthine contents in Korean native chicken -red-brown line (KNC-R Line). Methods: A total sample of 284 (males, n = 127; females n = 157) and 230 (males, n = 106; females, n = 124) aged of 10 weeks old KNC-R line was used for genotyping of DUSP8 and IGF2 genes, respectively. One SNP (rs313443014 C>T) in DUSP8 gene and two SNPs (rs315806609A/G and rs313810945T/C) in IGF2 gene were used for genotyping by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and KASP methods, respectively. The Two-way analysis of variance of the R program was used to associate DUSP8 and IGF2 genotypes with nucleotide contents in KNC-R chickens. Results: The DUSP8 (rs313443014 C>T) was polymorphic in KNC-R line and showed three genotypes: CC, CT, and TT. The IGF2 gene (rs315806609A/G and rs313810945T/C) was also polymorphic and had three genotypes per SNP, including GG, AG, and AA for the SNP rs315806609A/G and genotypes: CC, CT, and TT for the SNP rs313810945T/C. Association resulted into a strong significant association (p<0.01) with IMP, inosine, and hypoxanthine. Moreover, the significant effect of sex (p<0.05) on nucleotide content was also observed. Conclusion: The SNPs in the DUSP8 and IGF2 genes might be used as genetic markers in the selection and production of chickens with highly flavored meat.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2020.12a
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• pp.63-63
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• 2020

Rice stripe virus (RSV) disease is one of the major constraints in rice production, transmitted by the small brown planthopper (SBPH; Laodelphax striatellus). Upon RSV infection, plants develop typical symptoms, which include chlorosis and weakness of newly emerged leaves, white and yellow spots, stripe on leaves, and necrotic and wilting leaves, resulting in plant growth inhibition, oxidative damage that may culminate in programmed cell death (PCD) and plant death in severe epidemics. Although RSV-resistant quantitative trait loci (QTLs), Stv-a, Stv-b, and Stv-bi, were mapped using various resistant varieties, one RSV-resistant gene, OsSOT1, has been identified so far. In this study, we used the rice cultivar Zenith, known to carry Stv-b, to investigate novel RSV-genes through fine mapping. Therefore, we crossed Zenith (Donor parent, RSV resistant) with Ilpum (Recurrent parent, RSV susceptible) to fine-map using a BC₂F₂ population of 2100 plants. Chromosome segment introgression lines that were heterozygous at a different region were selected, two types of heterozygous lines showed an heterozygous genotype between Sid2 and Sid75 to Indel9 and RM6680. Interestingly, we identified qSTV11^Z region harboring Stv-b, covering about 171-kb region between the InDel markers Sid75 and Indel8. The localization of qSTV11^Z provides useful information that could be used for marker-assisted selection and determination of genetic resources in rice breeding.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2020.12a
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• pp.54-54
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• 2020

지구온난화에 따라 농업부문 신재생에너지의 중요성이 증대되고 있으며, 화본과 식물은 바이오에너지작물의 중요한 소재를 제공하고 있다. 화본과 식물의 기내대량증식연구의 일환으로 홍띠식물의 기내 재생 식물체의 유전적 안정성에 대한 기초자료를 제공할 목적으로 기내배양으로 재분화시킨 홍띠(Imperata cylindrica 'Rubra') 재분화 식물체 중 녹색체 재생식물체를 대상으로 ISSR 표지를 사용하여 유전적 안정성을 조사하였다. 재분화식물체는 MS (Murashige and Skoog, 1962)배지에 생장조절제를 첨가한 배지에서 배양하였다. 생장점 부위를 적출하여 캘러스를 유도하고(0.1 mg/L 2,4-D와 2 mg/L BA), 캘러스 증식(0.1 mg/L 2,4-D와 0.05 mg/L BA), 신초 재분화( 0.01 mg/L NAA와 2 mg/L BA) 후 MS배지에서 식물체를 양성하고 순화시켰다. 배양은 26±2℃, 25 µmol/m²/s, 14h/10h (day/night) 광조건 하에서 실시하였다. 재분화식물체는 홍띠 및 녹색 재분화식물체 2 종류로 나타났는데, 이는 생장점에서는 홍띠가 분화되었음에도 불구하고 생장점 주변조직에서 유래한 녹새체가 분화된 후 우세하게 자라서 녹색재생체가 우점하는 것으로 추정된다. ISSR 분석은 대조구로 모식물체 홍띠를(8개체), 재분화식물체는 녹색체 중, 1년간 노지포장에서 재배중인 녹색체(10개체)와 실험실내 화분에서 재배중인 시료를(10개체) 사용하였다. ISSR 밴드패턴을 비교한 결과, 재분화체는 실내포트 재배식물체 10.3%, 노지1년 재배식물체 8.3%로 대조구의 4.1%보다 유전적 다형성 비율이 2배 이상 높게 나타났다. 또한 재분화식물체들의 유전적 유사도를 평가하고 군집분석을 실시하였다.

• ALGAE
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• v.39 no.1
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• pp.43-50
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• 2024

Oomycetes are ubiquitous heterotrophs of considerable economic and ecological importance. Lately their diversity in marine environments has been shown to be greatly underappreciated and many lineages of intracellular holocarpic parasites, infecting micro- and macro-algae, remain to be fully described taxonomically. Among them, pathogens of marine red algae have been studied extensively as they infect important seaweed crops. Throughout the 20th century, most intracellular, holocarpic biotrophic oomycetes that infect red algae have been assigned to the genus Olpidiopsis Cornu. However, 18S rRNA sequencing of Olpidiopsis saprolegniae, the species considered the generitype for Olpidiopsis, suggests that this genus is not closely related to the marine pathogens and that the latter requires a nomenclatural update. Here, we compile and reanalyze all recently published 18S rRNA sequence data for marine holocarpic oomycetes, with a particular focus on holocarpic pathogens of red algae. Their taxonomy has been revised twice over the past four years, with suggestions to transfer them first into the genus Pontisma and then Sirolpidium, and into a monogeneric order, Pontismatales. We show however, that previously published topologies and the proposed taxa Pontisma, Sirolpidium, and Pontismatales are unsupported. We highlight that name changes that are unfounded and premature create confusion in interested parties, especially concerning pathogens of marine red algae that infect important seaweed crops. We thus propose that the names of these holocarpic biotrophic parasites of red algae are retained temporarily, until a supported topology is produced with more genetic markers to enable the circumscription of species and higher-level taxa.

• The Plant Pathology Journal
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• v.40 no.2
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• pp.171-191
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• 2024

Identification of Helicotylenchus species is very challenging due to phenotypic plasticity and existence of cryptic species complexes. Recently, the use of rDNA barcodes has proven to be useful for identification of Helicotylenchus. Molecular markers are a quick diagnostic tool and are crucial for discriminating related species and resolving cryptic species complexes within this speciose genus. However, DNA barcoding is not an error-free approach. The public databases appear to be marred by incorrect sequences, arising from sequencing errors, mislabeling, and misidentifications. Herein, we provide a comprehensive analysis of the newly obtained, and published DNA sequences of Helicotylenchus, revealing the potential faults in the available DNA barcodes. A total of 97 sequences (25 nearly full-length 18S-rRNA, 12 partial 28S-rRNA, 16 partial internal transcribed spacer [ITS]-rRNA, and 44 partial cytochrome c oxidase subunit I [COI] gene sequences) were newly obtained in the present study. Phylogenetic relationships between species are given as inferred from the analyses of 103 sequences of 18S-rRNA, 469 sequences of 28S-rRNA, 183 sequences of ITS-rRNA, and 63 sequences of COI. Remarks on suggested corrections of published accessions in GenBank database are given. Additionally, COI gene sequences of H. dihystera, H. asiaticus and the contentious H. microlobus are provided herein for the first time. Similar to rDNA gene analyses, the COI sequences support the genetic distinctness and validity of H. microlobus. DNA barcodes from type material are needed for resolving the taxonomic status of the unresolved taxonomic groups within the genus.

• The Korean Journal of Mycology
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• v.27 no.6 s.93
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• pp.394-398
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• 1999

Taxonomic and genetic analysis of Phytophthora species belonging to six different morphological groups (GI, GII, GIII, GIV, GV, GVI) was conducted using RAPD method. Amplified fragments ranged $0.3{\sim}3.2$ kb in their molecular weights. Among total of 145 bands, there were 109 polymorphic bands. Seven isolates of P. infestans showed high similarities of $0.92{\sim}0.99$, and P. infestans isolate 3 from potato showed similarities of $0.93{\sim}0.95$ compared with other P. infestans. Among isolates of P. capsici, similarities of $0.77{\sim}0.86$ were observed and they were grouped in 80% level. P. cinnamomi and P. cryptogea isolates which belonging to group GVI showed very similar RAPD fingerprinting pattern. Primers OPA-04, OPA-17, OPA-18, OPA-19, and OPB-12 showed high level of differences among the tested isolates in major bands and molecular weights. The similarity between the isolates was 0.67. P. megasperma and P. sojae in group GV showed similarity of 0.65. These two isolates showed big differences in single major band in reactions with primers OPA-08, OPA-17, and OPA-19. Phytophthora-specific and P. infestans-specific molecular markers were also selected with one of the random primers tested. In reaction with primer OPA-20, all the genus Phytophthora showed common band at 600 bp, and all the P. infestans isolates showed specific band at 680 bp. These markers can be useful for identification of Phytophthora speices or P. infestans. As a result, P. infestans isolated from tomato and/or potato can easily be differentiated from other Phytophthora species with this primer.

• Tuberculosis and Respiratory Diseases
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• v.50 no.6
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• pp.668-675
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• 2001

Background : Non-smalll lung cancer(NSCLC) develops as a result of the accumulation of multiple genetic abnormalities. Loss of heterozygosity(LOH) is one of the most frequent genetic alterations that is found in NSCLC, and the chromosomal regions that display a high rate of LOH are thought to harbor tumor suppressor genes(TSGs). This study was done to determine the frequency of LOH in 21q with the aim of identifying potential TSG loci. Method : Thirty-nine surgically resected NSCLCs were analysed. Patients peripheral lymphocytes were used as the source of the normal DNA. Five microsatellite Inarkers of 21q were used to study LOH : 21q21.1(D21S1432, and D21S1994); 21q21.2-21.3(D21S1442) ; 21q22.1(21S1445) ; and 21q22.2-22.3(D21S266). The fractional allelic loss(FAL) in a tumor was calculated as the ratio of the number of markers showing LOH to the number of informative markers. Result : LOH for at least one locus was detected in 21 of 39 tumors(53.8%). Among the 21 tumors with LOH, 5(21.8%) showed LOH at almost all informative loci. Although statistically not significant, LOH was found more frequently in squamous cell carcinomas(15 of 23, 65.2%) than in adenocarcinomas(6 of 16, 37.5%). In the squamous cell carcinomas the frequency of LOH was higher in stage II-III (80.0%) than in stage I (53.8%). The FAL value in squamous cell carcinomas($0.431{\pm}0.375$) was significantly higher than that found in adenocarcinomas($0.l92{\pm}0.276$). Conclusion : These results suggest that LOH on 21q may be involved in the development of NSCLC, and that TSG(s) that contribute to the pathogenesis of NSCLC may exist on 21q.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.91-91
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• 2017

Drought is the most serious abiotic stress limiting rice production. However, little progress has been made in the genetic analysis of drought tolerance, because it is a complex trait controlled by a number of genes and affected by various environmental factors. In here, we screened 218 rice genetic resources for drought tolerance at vegetative stage and selected 32 highly drought tolerant varieties in greenhouse. Under rain-fed conditions, Grain yield of Nagdong was decreased by 53.3% from 517 kg/10a to 241 kg/10a when compare to irrigation condition. By comparison, grain yield of Samgang was decreased by 23.6% from 550 kg/10a to 420 kg/10a. The variety Samgang exhibited strong drought tolerance and stable yield in rain-fed conditions and was selected for further study. To identify QTLs for drought tolerance, we examined visual drought tolerance (VDT) and relative water content (RWC) using a doubled haploid (DH) population consisted of 101 lines derived from a cross between Samgang (a drought tolerance variety) and Nagdong (a drought sensitive variety). Three QTLs for VDT were located on chromosomes 2, 6, and 11, respectively, and explained 41.8% of the total phenotypic variance. qVDT2, flanked by markers RM324 and S2016, explained 8.8% of the phenotypic variance with LOD score of 3.3 and an additive effect of -0.6. qVDT6 was flanked by S6022 and S6023 and explained 12.7% of the phenotypic variance with LOD score of 5.0 and an additive effect of -0.7. qVDT11, flanked by markers RM26765 and RM287, explained 19.9% of the phenotypic variance with LOD score of 7.1 and an additive effect of -1.0. qRWC11 was the only QTL for RWC to be identified; it was in the same locus as qVDT11. qRWC11 explained 19.6% of the phenotypic variance, with a LOD score of 4.0 and an additive effect of 9.7. To determine QTL effects on drought tolerance in rain-fed paddy conditions, seven DH lines were selected according to the number of QTLs they contained. Of the drought tolerance associated QTLs, qVDT2 and qVDT6 did not affect tiller formation, but qVDT11increased tiller number. Tiller formation was most stable when qVDT2 and qVDT11 were combined. DH lines with both of these drought tolerance associated QTLs exhibited the most stable tiller formation. These results suggest that qVDT11 is important for drought tolerance and stable tiller formation under drought stress condition in field.