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Characterization of exopolysaccharide-producing lactic acid bacteria from Taiwanese ropy fermented milk and their application in low-fat fermented milk

  • Ng, Ker-Sin;Chang, Yu-Chun;Chen, Yen-Po;Lo, Ya-Hsuan;Wang, Sheng-Yao;Chen, Ming-Ju
    • Animal Bioscience
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    • v.35 no.2
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    • pp.281-289
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    • 2022
  • Objective: The aim of this study was to characterize the exopolysaccharides (EPS)-producing lactic acid bacteria from Taiwanese ropy fermented milk (TRFM) for developing a clean label low-fat fermented milk. Methods: Potential isolates from TRFM were selected based on the Gram staining test and observation of turbid suspension in the culture broth. Random amplified polymorphic DNA-polymerase chain reaction, 16S rRNA gene sequencing, and API CHL 50 test were used for strain identification. After evaluation of EPS concentration, target strains were introduced to low-fat milk fermentation for 24 h. Fermentation characters were checked: pH value, acidity, viable count, syneresis, and viscosity. Sensory evaluation of fermented products was carried out by 30 volunteers, while the storage test was performed for 21 days at 4℃. Results: Two EPS-producing strains (APL15 and APL16) were isolated from TRFM and identified as Lactococcus (Lc.) lactis subsp. cremoris. Their EPS concentrations in glucose and lactose media were higher than other published strains of Lc. lactis subsp. cremoris. Low-fat fermented milk separately prepared with APL15 and APL16 reached pH 4.3 and acidity 0.8% with a viable count of 9 log colony-forming units/mL. The physical properties of both products were superior to the control yogurt, showing significant improvements in syneresis and viscosity (p<0.05). Our low-fat products had appropriate sensory scores in appearance and texture according to sensory evaluation. Although decreasing viable cells of strains during the 21-day storage test, low-fat fermented milk made by APL15 exhibited stable physicochemical properties, including pH value, acidity, syneresis and sufficient viable cells throughout the storage period. Conclusion: This study demonstrated that Lc. lactis subsp. cremoris APL15 isolated from TRFM had good fermentation abilities to produce low-fat fermented milk. These data indicate that EPS-producing lactic acid bacteria have great potential to act as natural food stabilizers for low-fat fermented milk.

Application of AFLPs to Phylogenetic Analysis of Aegilops (AFLPs에 의한 Aegilops의 계통발생학적 재평가)

  • Park, Yong-Jin;Shim, Jae-Wook
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.6
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    • pp.790-799
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    • 1997
  • Aegilops genus is known to include the donor species of the Band D genome of the bread wheat(ABD). An effort to establish a better strategy for phylogenetic relationships about Aegilops polyploids by AFLPs(Amplified Fragment Length Polymorphisms) was conducted using the 19 Aegilops sPP. and T. aestivum. The 207 polymorphic bands from the amplified products on the 6% acrylamide denaturing sequencing gels were obtained with the 7 AFLP primer combinations, and used to account for the genetic similarities and cluster analysis using NTSYS program. According to the genome analysis, the $M^h$-genome of Ae. heldreichii was estimated as an intermediate genome between the M-genome of Ae. comosa and N-genome of Ae. uniaristata and supposed to be incorporated in the establishing process of UM-genome as a possible diploid donor. And Ae. ventricosa(DN) was more close to Ae. umbellulata(U) than Ae. squarrosa(D). The close relationship between Ae. squarrosa and T. aestivum was perceived as a diploid donor of D-genome. As for the polyploid species, hexaploid Ae. triaristata was more closely related to Ae. columnaris rather than tetraploid Ae. triaristata. The clustered groups were, basically same to the previous Gihara's sections based on phenotypes and pairing analysis of interspecific hybrids. AFLP was evaluated as an efficient and powerful method in the genome evaluation of closely related species.

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Identification and Characterization of Polymorphic Microsatellite DNA Markers Using Next-generation Sequencing in Parapristipoma trilineatum (차세대 염기서열 분석법을 사용한 벤자리(Parapristipoma trilineatum)의 microsatellite 마커의 개발 및 유전학적 특성 분석)

  • Chun Mae Dong;Mi-Nan Lee;Jae Koo Noh;Jin Woo Park;Young-Ok Kim;Eun-Mi Kim
    • Journal of Life Science
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    • v.33 no.8
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    • pp.623-631
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    • 2023
  • This study was conducted to develop microsatellite markers in Parapristipoma trilineatum using next-generation sequencing. A total of 402,244,934 reads were generated on the Illumina Hiseq X Ten System, yielding 60,738,985,034 bp of sequences. The de novo assembly resulted in 1,320,995 contigs. A total of 952,326 contigs (0.016%) including 151 microsatellite loci were derived from the 1,320,995 contigs longer than 640 bp. A total of 34 primer sets were designed from the 151 microsatellite loci. As a result, 15 microsatellite loci were chosen and used for assuming population genetic parameters in the wild and farmed populations. The mean number of effective alleles was 12, ranging from 6 to 25. The observed heterozygosity (HO) and the expected heterozygosity (HE) ranged between 0.530 and 0.873, with an average of 0.750, and from 0.647 to 0.895, with an average of 0.793, respectively. According to these results, the developed set of 15 microsatellite markers is expected to be useful for the analysis of genetic characteristics in the population of P. trilineatum in Korea. There are requirements now for further genetic information, fishery resource management, breeding guidelines, support with the selection of breeds and studies on the effects of release, all of which will improve species conservation, and through future research, we aim to offer genetic foundational data with that goal.

Characteristics of Dioscorea alata L. Introduced from Tropical and Subtropical Regions (도입 마(Dioscorea alata L.)의 특성 분석)

  • Chang, K.J.;Yoo, K.O.;Park, C.H.;Park, J.I.;Hong, K.H.;Park, J.H.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.3 no.1
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    • pp.48-69
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    • 2001
  • A lot of clones of the genus Dioscorea have been introduced from some tropical and subtropical regions since 1997. In 33 clones of water yams (Dioscorea alata L.), some morphological characteristics were investigated at the field. Variation ranges of the total weight and tuber number per stump were within the ranges from 90 to 2,147 g with an average of 610 g ; and 1.3-4.7 with an average of 2.8, respectively. The color tones observed on the tuber-flesh were sorted into 3 color-categories, i.e., white, pale brown and pale purple, and those on leaves were sorted into 3 color-categories, i.e., green, heavy green and purplish green. Intraspecific genetic relationship of 19 variation types of the Yam classified by their external morphological characteristics such as leaf and tuber shape was assessed by DNA using random and specific primers. Twenty two out of 113 primers (100 random[10-mer] primers, two 15 mer [M13 core sequence, and (GGAT)4 sequence]) had been used in PCR-amplification. Only 12 primers, however, were successful in DNA amplification in all of the analyzed plants, resulting in 93 randomly and specifically amplified DNA fragments. The analyzed taxa showed very high polymorphisms(69 bands, 71.0%), allowing individual taxon to be identified based on DNA fingerprinting. Monomorphic bands among total amplified DNA bands of each primer was low under the 50%. Similarity indices between accessions were computed from PCR(polymerase chain reaction) data, and genetic relationships among intraspecific variations were closely related at the levels ranging from 0.66 to 0.90.

Development of SCAR Markers for the Discrimination of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma based on the RAPD (RAPD 분석을 통한 대황(大黃)과 종대황(種大黃) 감별용 SCAR 유전자 마커 개발)

  • Moon, Byeong-Cheol;Lee, Young-Mi;Chun, Jin-Mi;Lee, A-Young;Yoon, Tae-Sook;Cheon, Myeong-Sook;Choo, Byung-Kil;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.115-120
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    • 2009
  • Objectives : Due to the morphological similarity and frequent occurrence of intermediate forms as well as morphological variations of aerial part, the correct identification between Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma is very difficult. To develop a reliable method for correct identification and improving the quality standards of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma, we analyzed RAPD and developed SCAR marker. Methods : To amplify target DNA at the genomic level, 32 Operon 10-mer random primers were applied with four Rheum species, R. officinale, R. palmatum, R. tanguticum and R. undulatum. The nucleotide sequences were determined and species-specific primers were prepared depending on the species-specific RAPD amplicons after subcloned into the pGEM-Teasy vector. To develop the SCAR markers, species-specific PCR amplification and multiplex-PCR were carried out using the single species-specific primer pairs and combinations of them, respectively. Results : We used RAPD analysis of four Rheum plant species to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed two SCAR markers that amplified 314 bp and 390 bp DNA fragments in only R. undulatum but not in R. officinale, R. palmatum, R. tanguticum and R. undulatum, for distinguishing Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. Conclusions : These genetic markers can be used for the efficient discrimination of plants species and commercial herbal medicines between Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma, to ultimately prevent indiscriminate distribution and prescription of these herbal medicines.

The Role of CYP2B6*6 Gene Polymorphisms in 3,5,6-Trichloro-2-pyridinol Levels as a Biomarker of Chlorpyrifos Toxicity Among Indonesian Farmers

  • Liem, Jen Fuk;Suryandari, Dwi A.;Malik, Safarina G.;Mansyur, Muchtaruddin;Soemarko, Dewi S.;Kekalih, Aria;Subekti, Imam;Suyatna, Franciscus D.;Pangaribuan, Bertha
    • Journal of Preventive Medicine and Public Health
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    • v.55 no.3
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    • pp.280-288
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    • 2022
  • Objectives: One of the most widely used pesticides today is chlorpyrifos (CPF). Cytochrome P450 (CYP)2B6, the most prominent catalyst in CPF bioactivation, is highly polymorphic. The objective of our study was to evaluate the role of CYP2B6*6, which contains both 516G>T and 785A>G polymorphisms, in CPF toxicity, as represented by the concentration of 3,5,6-trichloro-2-pyridinol (TCPy), among vegetable farmers in Central Java, Indonesia, where CPF has been commonly used. Methods: A cross-sectional study was conducted among 132 vegetable farmers. Individual socio-demographic and occupational characteristics, as determinants of TCPy levels, were obtained using a structured interviewer-administered questionnaire and subsequently used to estimate the cumulative exposure level (CEL). TCPy levels were detected with liquid chromatography-mass spectrometry. CYP2B6*6 gene polymorphisms were analyzed using a TaqMan® SNP Genotyping Assay and Sanger sequencing. Linear regression analysis was performed to analyze the association between TCPy, as a biomarker of CPF exposure, and its determinants. Results: The prevalence of CYP2B6*6 polymorphisms was 31% for *1/*1, 51% for *1/*6, and 18% for *6/*6. TCPy concentrations were higher among participants with CYP2B6*1/*1 than among those with *1/*6 or *6/*6 genotypes. CYP2B6*6 gene polymorphisms, smoking, CEL, body mass index, and spraying time were retained in the final linear regression model as determinants of TCPy. Conclusions: The results suggest that CYP2B6*6 gene polymorphisms may play an important role in influencing susceptibility to CPF exposure. CYP2B6*6 gene polymorphisms together with CEL, smoking habits, body mass index, and spraying time were the determinants of urinary TCPy concentrations, as a biomarker of CPF toxicity.

Improvement of Pre-harvest Sprouting Resistance in Korean japonica Varieties through a Precision Marker-based Breeding

  • Kamal Bhattarai;Patricia Izabelle Lopez;Sherry Lou Hechanova;Ji-Ung Jeung;Hyun-Sook Lee;Eok-Keun Ahn;Ung-Jo Hyun;Jong-Hee Lee;So-Myeong Lee;Jose E. Hernandez;Sung-Ryul Kim
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.269-269
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    • 2022
  • Pre-harvest sprouting (PHS) on rice panicles is getting problematic in recent several years in Korea due to climate changes such as high temperature and more frequent typhoons during harvesting season. PHS negatively affects grain quality severely and also yield. Genetic improvement of Korean varieties (Oryza sativa ssp. japonica) through a marker assisted-backcross breeding (MAB) with the known PHS resistant genes must be one of ideal solutions. However, the final breeding products of MAB occasionally exhibit unwanted traits, especially the cross between genetically distant parents. This might be caused by linkage drag and/or presence of the gene-unlinked donor introgressions, resulting that the final products could not be released to the farmers. The major PHS resistance gene, Sdr4 (Seed dormancy 4) originated from an indica cultivar, Kasalath was selected as a donor gene. In order to avoid unexpected phenotypes in the breeding products, we performed a precision marker-based breeding (PMBB) consisting of foreground, recombinant, and background selections (FS, RS, and BS) which aim to develop 'single small introgression lines' (~100 kb introgression). Korean varieties (Ilpum and Gopum) were crossed with Kasalath. We developed Sdr4-allele specific markers for FS and a set of polymorphic flanking markers near the Sdr4 (-350kb and +420kb) for RS. To minimize linkage drag, the small introgression (< 125kb) containing Sdr4 was selected in Ilpum background (BC2F4) through 1st RS with ~1,200 F2 or BC1F2 plants (one side trimmed) and then 2nd RS with ~1,000 progenies from the 1st RS selected plants (another side trimmed). After RS, the selected lines were genotyped by using Infinium 7K SNP chip to detect other donor introgressions and the lines were backcrossed. Currently BS is on-going from the backcross-derived progenies with BS markers to remove residual introgressions. During the PMBB process, genetic effect of Sdr-4-Kasalath allele was confirmed in Ilpum and Gopum backgrounds by PHS phenotyping using the segregating BC2F3 or BC1F4 materials. The Sdr4 PMBB lines in Ilpum background (< 125kb introgression) will be valuable genetic resources to improve PHS resistance in modem popular temperate japonica varieties.

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Comparison of Antioxidant Activity and Flavor Effect According to Processing Method of Red ginseng and Herbal Medicine (홍삼 및 생약재의 가공방법에 따른 항산화활성 특성 및 향미 효과 비교)

  • Hyun Kyoung Kim;Ho Tae Kim;Pil Jae Lee
    • The Journal of the Convergence on Culture Technology
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    • v.9 no.6
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    • pp.1007-1016
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    • 2023
  • In this study, the effect of functional substances such as polyphenols and flavonoids contained in large quantities in red ginseng and herbal medicines on the antioxidant activity properties and flavor effect were investigated by increasing the content and activity of functional ingredients by convert red ginseng through a steaming heat-drying process compared to the traditional dry processing method of ginseng and herbal medicines. According to the experimental results, the addition of pre-heat treatment significantly increased antioxidant properties such as DPPH radical scavenging ability, polyphenols, and flavonoids. In addition, during the steaming and heat-dried red ginseng manufacturing process, the amino-carbonyl browning reaction was promoted, resulting in increased brownness and a savory flavor. However, the content of ginsenosides, the main medicinal ingredient in red ginseng, became the main cause of the bitter and harsh taste. In addition, the bitter and harsh taste of red ginseng has been significantly improved by roasting and producing powder, but on the other hand, Radix Angelicae sinensis, polymorphic angelica and Peony, which are used as main medicinal ingredients in oriental medicine for nutritional tonic prescriptions such as 10 herbal medicine and 4 herbal medicine have a very strong herbal medicine-specific flavor and have a bitter and harsh taste. It is so strong that in order to use it as an instant extraction material, it was reviewed that a steaming and heating manufacturing method was needed during the manufacturing process.

G-, C-, and NOR-banding of Korean Native Pig Chromosomes (한국재래돼지의 G-, C-, 및 NOR-banding)

  • Sohn, S.H.;Kweon, O.S.;Baik, K.H.;Jung, W.;Cho, E.J.;Kang, M.Y.
    • Journal of Animal Science and Technology
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    • v.45 no.6
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    • pp.901-910
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    • 2003
  • Using the G-, C-, and NOR-banding techniques, a karyotyping for Korean Native Pig was performed. Blood samples were collected from 50 male Korean Native Pigs that had been bred at the National Livestock Research Institute and then blood cells were prepared from in vitro cultures followed by karyotyping; G-, C-, and NOR-banding patterns of metaphase chromosomes were analyzed. The karyotype of Korean Native Pig is 38, XX or XY which consists of 5 pairs of submetacentric chromosomes(Group I), 2 pairs of acrocentric chromosomes with short p-arm(Group II), 5 pairs of medium metacentric chromosomes(Group III), 6 pairs of acrocentric chromosomes(Group IV) and metacentric X and Y sex chromosomes. On GTG-banding, the Korean Native Pig exhibited a typical and identical banding pattern in each homologous chromosomes. Overall chromosomal morphology and positions of typical landmarks of the Korean Native Pig were virtually identical to those of Committee for the Standardized Karyotype of the Domestic Pig(CSKDP). However, numbers of G-bands of the Korean Native Pig chromosomes were more than those of CSKDP. In chromosomes 1, 3, 5, 6, 7, 8, 13, 14, 15, 16, 17, 18 and X, the Korean Native Pig exhibited more separated bands as compared with CSKDP. In C-banding patterns, although the quantity of heterochromatin was variable in each chromosome, most of the Korean Native Pig chromosomes had heterochromatic C-bands on centromeres. However, the heterochromatic C-band was constantly observed on the whole Y chromosome. In AgNOR staining, the NORs were located at centromeres on the chromosomes 8 and 10. The number of NORs per metaphase ranged from 2 to 4 giving a mean value of 2.13. The number of NORs were distributed on all chromosome pair 10 but not on chromosome 8. The sizes of NORs were also differed between homologous chromosomes 8. Numbers of NORs of Korean Native Pig were significantly higher than those of Yorkshire. The pattern of pig NORs was polymorphic in breeds, individuals and cells, especially on chromosome 8.

Association Analysis of MUC5AC Promoter Polymorphism with Asthma (MUC5AC 프로모터의 유전자 다형성과 천식과의 연관성)

  • Han, Seon-Sook;Sung, Ji Hyun;Lee, Mi-Eun;Lee, Seung-Joon;Lee, Sung Joon;Kim, Woo Jin
    • Tuberculosis and Respiratory Diseases
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    • v.63 no.3
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    • pp.235-241
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    • 2007
  • Background: Airway mucus hypersecretion plays an important role in the pathogenesis of asthma, and is associated with the induction of MUC5AC expression in airway secretion. The MUC5AC gene is highly polymorphic; however, there are few studies about the association between the polymorphisms of the MUC5AC gene and asthma susceptibility or asthma phenotypes. We have investigated the association of MUC5AC promoter polymorphisms with the risk of asthma and asthma phenotypes. Methods: We determined the genotypes of the MUC5AC promoter (-1274G>A) in 78 asthma patients and in 78 age, sex-matched control individuals in the Korean population. Genomic DNAs from blood were analyzed by PCR and RFLP (restriction fragment length polymorphism) determination. We examined $FEV_1$, total eosinophil count, serum IgE level, $PC_{20}$ and the presence of atopy (by a skin test) in asthma patients. Results: The mean age of the patients was $47.7{\pm}16.1$ years and 38.5% were men, and the mean $FEV_1$ was $84.4{\pm}22.3%$ of predicted in the asthma patients. The -1274G>A polymorphism of the MUC5AC promoter in asthma patients was not significantly different as compared with normal individuals (GG 57.7%, AG 34.6% and AA 7.7% in asthma patients vs. GG 56.4%, AG 38.5% and AA 5.1% in control subject, p = 0.752, Cod). Several clinical parameters in asthma patients such as $FEV_1$, total eosinophil count, serum IgE level, $PC_{20}$ and the presence of atopy, were not associated with the -1274G>A polymorphism of the MUC5AC promoter. Conclusion: The -1274G>A single nucleotide polymorphism (SNP) frequency of the MUC5AC promoter was not associated with asthma in a Korean population.