• Animal Bioscience
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• v.37 no.5
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• pp.929-943
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• 2024

Objective: The present study was executed to explore the molecular mechanism of fibroblast growth factor 10 (FGF10) gene in bovine adipogenesis. Methods: The bovine FGF10 gene was overexpressed through Ad-FGF10 or inhibited through siFGF10 and their negative control (NC) in bovine adipocytes, and the multiplicity of infection, transfection efficiency, interference efficiency were evaluated through quantitative real-time polymerase chain reaction, western blotting and fluorescence microscopy. The lipid droplets, triglycerides (TG) content and the expression levels of adipogenic marker genes were measured during preadipocytes differentiation. The differentially expressed genes were explored through deep RNA sequencing. Results: The highest mRNA level was found in omasum, subcutaneous fat, and intramuscular fat. Moreover, the highest mRNA level was found in adipocytes at day 4 of differentiation. The results of red-oil o staining showed that overexpression (Ad-FGF10) of the FGF10 gene significantly (p<0.05) reduced the lipid droplets and TG content, and their down-regulation (siFGF10) increased the measurement of lipid droplets and TG in differentiated bovine adipocytes. Furthermore, the overexpression of the FGF10 gene down regulated the mRNA levels of adipogenic marker genes such as CCAAT enhancer binding protein alpha (C/EBPα), fatty acid binding protein (FABP4), peroxisome proliferator-activated receptor-γ (PPARγ), lipoprotein lipase (LPL), and Fas cell surface death receptor (FAS), similarly, down-regulation of the FGF10 gene enriched the mRNA levels of C/EBPα, PPARγ, FABP4, and LPL genes (p<0.01). Additionally, the protein levels of PPARγ and FABP4 were reduced (p<0.05) in adipocytes infected with Ad-FGF10 gene and enriched in adipocytes transfected with siFGF10. Moreover, a total of 1,774 differentially expressed genes (DEGs) including 157 up regulated and 1,617 down regulated genes were explored in adipocytes infected with Ad-FGF10 or Ad-NC through deep RNA-sequencing. The top Kyoto encyclopedia of genes and genomes pathways regulated through DEGs were the PPAR signaling pathway, cell cycle, base excision repair, DNA replication, apoptosis, and regulation of lipolysis in adipocytes. Conclusion: Therefore, we can conclude that the FGF10 gene is a negative regulator of bovine adipogenesis and could be used as a candidate gene in marker-assisted selection.

• Korean Journal of Plant Resources
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• v.34 no.5
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• pp.443-450
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• 2021

Peach flesh color is commercially important criteria for classification and has implications for nutritional quality. To breed new yellow-fleshed peach cultivar many cross seedlings and generations should be maintained. Therefore it is necessary to develop early selection molecular markers for screening cross seedlings and germplasm with economically important traits to increase breeding efficiency. For the comparison of transcription profiles in peach varieties with a different flesh color expression, two cDNA libraries were constructed. Differences in gene expression between yellow-fleshed peach cultivar, 'Changhowon Hwangdo' and white-fleshed peach cultivar, 'Mibaekdo' were analyzed by next-generation sequencing (NGS). Expressed sequence tag (EST) of clones from the two varieties was selected for nucleotide sequence determination and homology searches. Putative single nucleotide polymorphisms (SNPs) were screened from peach EST contigs by high resolution melting (HRM) analysis, SNP ID ppa002847m:cds and ppa002540m:cds displayed specific difference between 17 yellow-fleshed and 21 white-fleshed peach varieties. The SNP markers for distinguishing yellow and white fleshed peach varieties by HRM analysis offers the opportunity to use early selection. This SNP markers could be useful for marker assisted breeding and provide a good reference for relevant research on molecular mechanisms of color variation in peach varieties.

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.193-201
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• 2001

This study was designed to examine the factors affecting in fertilization and development of embryos in vitro, and to examine whether zone drilling by laser irradiation can improve the hatching rate of IVF embryos from DNA marker-proved Hanwoo. DNA markers related to marbling score were identified using DNA fingerprinting with Ml3 probe and restriction enzyme Hae III. Oocytes were aspirated from immature ovarian follicles using a combined method of rectal ovarian-palpation and transvaginal ultrasound-guidance(6.5MHz) under local anesthesia. The aspirated oocytes were washed twice with fresh D-PBS containing 5% FBS and were rewashed 4 to 5 times with TCM-199 containing 5% FBS. A morphological grade of I to IV was assigned to each oocyte. Data were analyzed using the GLM procedure of SAS. Sperm separation methods did not have any significant effect on cleavage or developmental abilities of IVF embryos. Significantly(P<0.05) higher cleavage rate was observed in embryos from GI(60.0%, 3/5), GII(69.2%, 18/26) and GIII(62.1%, 59/95) compared to embryos from GIV oocytes(36.2%, 25/69). And the developmental rate to blastocyst stage was higher(P<0.05) in embryos from GI(33.3%, 1/3) and GII oocytes(38.9%, 7/18) than those from GIII(16.9%,10/59) and GIV oocytes(4.0%, 1/25). There was no significant difference in development of IVF embryos to blastocyst by media for in vitro culture. Proportion of hatched blastocyst was significantly(P<0.05) higher in embryos received zona drilling by laser than those of non-drilled.

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.183-191
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• 2001

This study was designed to establish the superior method for IVF embryos from DNA marker-proved Hanwoo cattle. DNA markers related to marbling score were identified using DNA fingerprinting with Ml3 probe and restriction enzyme Hae III. Oocytes were aspirated from unstimulated. immature ovarian follicles using a combined method of rectal ovarian-palpation and transvaginal ultrasound-guidance(6.5MHz) under local abesthesia. The aspirated oocytes were washed twice with fresh D-PBS containing 5% FBS and were rewashed 4 to 5 times with TCM-199 containing 5% FBS. A morphological grade of I to IV was assigned to each oocyte. Data were analysed using the GLM procedure of SAS. Mean number of follicles identified on ultrasound was 5.5 $\pm$2.9 in right and 4.3 $\pm$2.8 in left ovaries, respectively. The highest follicles(16.6$\pm$2.6) were found in 5101 cow compared to others. Recovery rate of follicular oocytes in individual cow was highest in 5101 cow with 89.3% in > 2mm and 94.0% in $\leq$ 2mm follicles. Total recovery rate was significantly(P<0.01) higher in $\leq$ 2mm(85.7%, 130/154) than > 2mm follicles(74.2%, 201/271). Significantly more oocytcs of Grade IV were recovered from > 2mm follicles. Mean number follicles recovered was 4.8$\pm$3.7. 3.0$\pm$3.4 and 0.3$\pm$0.6 in $\leq$2mm, 2~6mm and $\geq$6mm follicles. respectively. Our results imply that the more fertilizablc oocytes can be recovered from invisible-immature follicles by the combination of simultaneous rectal ovarian-palpation and ultrasound-guided approach in Hanwoo cattle.

• Korean Journal of Breeding Science
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• v.40 no.4
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• pp.408-413
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• 2008

Rice blast resistance is considered one of the most important traits in rice breeding and the disease, caused by Magnaporthe grisea Barr, has brought significant crop losses annually. Moreover, breakdown of resistance normally occurs in two to five years after cultivar release, thus a more durable resistance is needed for better control of this disease. We developed a new variety, Keumo3, which showed strong resistance to leaf blast. It was tested in 2003 to 2007 at fourteen blast nursery sites covering entire rice-growing regions of South Korea. It showed resistance reactions in 12 regions and moderate in 2 regions without showing susceptible reactions. Durability test by sequential planting method indicated that this variety had better resistance. Results showed that Keumo3 was incompatible against 19 blast isolates with the exception of KI101 by artificial inoculation. To understand the genetic control of blast resistance in rice cultivar Keumo3 and facilitate its utilization, recombinant inbred lines (RIL) consisting of 290 F5 lines derived from Akidagomachi/Keumo3 were analyzed and genotyped with Pizt InDel marker zt56591. The recombination value between the marker allele of zt56591 and bioassay data of blast nursery test was 1.1%. These results indicated that MAS can be applied in selecting breeding populations for blast resistance using zt56591 as DNA marker.

• 한국균학회소식:학술대회논문집
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• 2015.05a
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• pp.33-33
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• 2015

Clubroot disease is the major threat to the production of Chinese cabbage (Brassica rapa L.) in Japan. Although the breeding of the clubtoot resistant (CR) cultivars is one of the most efficient ways to control this disease, the CR cultivars do not always have effects due to the breakdown of resistance. Therefore, it is necessary to develop the breeding strategy to accumulate multiple CR genes in a single cultivar effectively. We have identified two incomplete dominant CR loci, Crr1 and Crr2, which are originated from the European CR turnip Siloga. To investigate the effectiveness of marker-assisted selection (MAS) for CR breeding, the inbred line with Crr1 and Crr2 was crossed with parental lines of the existing CR $F_1$ cultivar of Chinese cabbage, followed by 5 times of MAS and backcrossing. The $F_1$ derived from a cross between the resulting parental lines improved the clubroot resistance as expected and had the same morphological characters as the original $F_1$ cultivar. We have shown that the Crr1 locus comprised two loci: Crr1a, which by itself conferred resistance to the mild isolate; and Crr1b, which had a minor effect, but was not required for Crr1a-mediated resistance. Further genetic analysis suggested that Crr1b was necessary to acquire resistance to the more virulent isolate in combination with Crr2. Molecular characterization of Crr1a encoding TIR-NB-LRR class of R protein revealed that there were at least 4 alleles in Japanese CR cultivars of Chinese cabbage. PCR analysis with Crr1a-specific markers demonstrated that the functional alleles were predicted to be present in European CR turnips, Debra and 77b besides Siloga, whereas rarely in Japanese CR cultivars, indicating that Crr1a is an useful source to improve the resistance of Chinese cabbage cultivars.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.11
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• pp.1555-1559
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• 2003

The objective of the present study was to explore associations between five microsatellites linked to $Fec^B$ and $FecX^I$ genes and litter size in Small Tail Han sheep. The polymorphisms of five microsatellite loci, OarAE101, BM1329, BMS2508, TGLA54 and TGLA68 were detected in 244 ewes of Small Tail Han sheep. Analysis of association between three microsatellite loci (BMS2508, BM1329 and OarAE101) located in the 10 cM region covering the $Fec^B$ gene (Booroola gene) and litter size in Small Tail Han sheep indicated that BMS2508 had significant effect on litter size in the second parity (p<0.05), but no significant effect on litter size in the first parity (p>0.05), while the other two microsatellite loci had no significant effect on litter size in both the first and the second parity in Small Tail Han sheep (p>0.05). At microsatellite locus BMS2508, least squares means in the second parity of genotypes 101/111 and 99/109 were significantly higher than those of genotypes 99/99, 99/101, 99/111 and 99/115 (p<0.05); least squares mean in the second parity of genotype 101/111 was significantly higher than that of genotypes 109/111 and 111/111 (p<0.05). Results of this study also indicated that two microsatellite loci (TGLA54 and TGLA68) that confined the 28.7 cM region covering the $FecX^I$ gene (Inverdale gene) did not affect litter size in both the first and the second parity in Small Tail Han sheep significantly (p>0.05). The information found in the present study is very important for improving the reproductive performance in sheep breeds by marker assisted selection.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.4
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• pp.422-427
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• 1998

The gene and genotypic frequencies of ${\kappa}$-casein (${\kappa}$-CN), ${\beta}$-lactoglobulin (${\beta}$-LG), growth hormone (bGH) and prolactin (bPRL) loci in Korean cattle were investigated using PCR-RFLP analyses. Genomic DNA samples were obtained from 290 cows and 30 AI bulls. In both cows and bulls, the most predominant genotypes of ${\kappa}$-CN, ${\beta}$-LG, bGH and bPRL loci were AB, BB, AA and AA, respecitively. The frequencies of A and B alleles for ${\kappa}$-CN locus were .612 and .388 for cows and .567 and .433 for bulls. The respective frequencies of A and B alleles for ${\beta}$-LG locus were .153 and .847 in cows and .217 and .783 in bulls. The frequencies of A and B alleles for bGH locus were .769 and .231 in cows and .784 and .216 in bulls, respectively. The frequencies of A and B alleles for bPRL locus were .678 and .322 for cows and .767 and .233 for bulls. Differences in frequencies of these alleles were not significant between cows and bulls at all loci examined. If the DNA polymorphisms of these candidate genes are associated with economically important traits, they could serve as genetic markers for genetic improvement in future marker-assisted selection programs in Korean cattle.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.4
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• pp.484-488
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• 2008

We investigated the effect of the prolactin receptor gene (PRLR) on total number of born (TNB), number of born alive (NBA) and number of weaned (NW) piglets in Large White (LW), White Meaty (WM) and Landrace (L) sows from six Slovak breeding farms. The frequency of A allele was 0.48, 0.49 and 0.47 in LW, WM and L, respectively. We found numerous highly significant effects of PRLR locus on TNB ($p{\leq}0.01$; $p{\leq}0.05$) in all tested breeds. The most marked difference of +$1.31{\pm}0.45pigs/L$ was found between AA and BB genotypes in WM. Within the other breeds the difference between the homozygous genotypes reached up to +$0.94{\pm}0.3$ and +$1.21{\pm}0.19$ pigs per litter in LW and L, respectively. We also identified significant differences between AA and AB genotypes related to TNB in L. Similarly NBA, as well as NW traits were significantly affected ($p{\leq}0.01$; $p{\leq}0.05$) by the genotype just in LW and L. The homozygous genotype AA was favourable in all breeds and traits. Our results showed the possibility of PRLR utilization in marker-assisted selection within breeding programs to increase reproductive traits of pigs in Slovakia.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.12
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• pp.1651-1659
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• 2011

This study was conducted to detect quantitative trait loci (QTL) for thirteen growth and meat quality traits in pigs by combing QTL experimental populations. Two F2 reference populations that were sired by Korea native pig (KNP) and dammed by Landrace (LN) or Yorkshire (YK) were generated to construct linkage maps using 123 genetic markers (mostly microsatellites) and to perform QTL analysis on porcine chromosomes (SSCs) 1, 2, 3, 6, 7, 8, 9, 11, 13, 14, and 15. A set of line-cross models was applied to detect QTL, and a series of lack-of-fit tests between the models was used to characterize inheritance mode of QTL. A total of 23, 11 and 19 QTL were detected at 5% chromosome-wise level for the data sets of KNP${\times}$LN, KNP${\times}$YK cross and joint sets of the two cross populations, respectively. With the joint data, two Mendelian expressed QTL for live weight and cooking loss were detected on SSC3 and SSC15 at 1% chromosome-wise level, respectively. Another Mendelian expressed QTL was detected for CIE a on SSC7 at 5% genome-wise level. Our results suggest that QTL analysis by combining data from two QTL populations increase power for QTL detection, which could provide more accurate genetic information in subsequent marker-assisted selection.