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The Studies of Activity of Retrotransposon(Tos17) according to Tissue Culture Periods in Rice(Oryza sativa L.) (벼 조직배양 기간에 따른 retrotransposon(Tos17)의 활성에 관한 연구)

  • Yang, Hee-Eun;Fang, Yilan;Shin, Young-Boum;Lee, Boung-Jin;Hong, Soon-Kwan
    • Korean Journal of Plant Resources
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    • v.20 no.5
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    • pp.389-397
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    • 2007
  • Using the active-increment of Tos17 copies in the genome of Oryza sativa L., there were many studies about induction and selection of new mutants. This study mainly focuses on the induction for retrotransposon(Tos17) activity in the callus of Ilpumbyeo(Oryza sativa L.) according to varied culture period and condition. The objectives of this study are obtaining various mutants($M_1$) through plant regeneration, identification of the mutation relation with Tos17, and subsequent phenotyping of the mutants($M_2,\;M_3$). A total of 371 $M_1$ mutants was obtained. The degree of Tos17 activity obtained regeneration plants with each different culture period was evaluated by Southern blot analysis. The result showed that control Ilpumbyeo rice has 5 numbers of copies and the band numbers obtained 7, 8, 9.5, 12, 6, 13.5, 17.5 from culture period of 1, 2, 3, 5, 6, 7, 8 month, respectively. In this study, the result showed that most effectual culture period for activity of Tos17 in Ilpumbyeo rice is 5 month. Hereafter, collections and analysis of various recombination plants will act on an important factor in multiplication and preservation of $M_2$ and $M_3$ generation. And an urgent and important subject is a development of screening method for selection of diverse mutants.

Development of molecular markers for varietal identification of Brassica juncea on the basis of the polymorphic sequence of ITS regions and MITE families (갓 (Brassica juncea) 품종구분을 위한 ITS 영역 및 MITE Family 정보를 이용한 분자표지 개발)

  • Yang, Kiwoung;Yi, Go-eun;Robin, Arif Hasan Khan;Jeong, Namhee;Lee, Yong-Hyuk;Park, Jongin;Kim, Hoyteak;Chung, Mi-Young;Nou, Ill-Sup
    • Horticultural Science & Technology
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    • v.34 no.2
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    • pp.305-313
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    • 2016
  • Brassica juncea (2n = 4x = 36, AABB genome, 1,068 Mb) is a U's triangle species and an amphidiploid derivative of B. rapa and B. nigra. Fifteen varieties were used to study the ITS (internal transcribed spacer) regions of ribosomal DNA and MITEs (miniature inverted-repeat transposable elements) with a view of developing specific molecular markers. ITSs and MITEs are an excellent resource for developing DNA markers for genomics and evolutionary studies because most of them are stably inherited and present in high copy numbers. The ITS (ITS1 and ITS2) sequence was compared with the consensus sequence of B. rapa and B. nigra. Variation in ITS1 created two separate groups among 15 varieties, with 10 varieties in one group and 5 in the other. Phylogenetic analysis revealed two major clusters for those 10 and 5 varieties. Among the 160 different MITE primers used to evaluate the selected 15 varieties of B. juncea, 70 were related to the Stowaway, 79 to the Tourist, 6 to the hAT, and 5 to the Mutator super-families of MITEs. Of 160 markers examined, 32 were found to be polymorphic when fifteen different varieties of B. juncea were evaluated. The variety 'Blackgat' was different from the other mustard varieties with respect to both phenotype and genotype. The diversity of 47 additional accessions could be verified using eight selected molecular markers derived from MITE family sequences. The polymorphic markers identified in this study can be used for varietal classification, variety protection, and other breeding purposes.

Microbial Community Composition Associated with Anaerobic Oxidation of Methane in Gas Hydrate-Bearing Sediments in the Ulleung Basin, East Sea (동해 울릉분지 가스 하이드레이트 매장 지역의 메탄산화 미생물 군집 조성 및 분포)

  • Cho, Hyeyoun;Kim, Sung-Han;Shin, Kyung-Hoon;Bahk, Jang-Jun;Hyun, Jung-Ho
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.20 no.1
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    • pp.53-62
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    • 2015
  • To elucidate the microbial consortia responsible for the anaerobic methane oxidation in the methane hydrate bearing sediments, we compared the geochemical constituents of the sediment, the rate of sulfate reduction, and microbial biomass and diversity using an analysis of functional genes associated with the anaerobic methane oxidation and sulfate reduction between chimney site (UBGH2-3) on the continental slope and non-chimney site (UBGH2-10) on the basin of the Ulleung Basin. From the vertical profiles of geochemical constituents, sulfate and methane transition zone (SMTZ) was clearly defined between 0.5 and 1.5 mbsf (meters below seafloor) in the UBGH2-3, and between 6 and 7 mbsf at the UBGH2-10. At the UBGH2-3, the sulfate reduction rate (SRR) in the SMTZ exhibited was appeared to be $1.82nmol\;cm^{-3}d^{-1}$ at the depth of 1.15 mbsf. The SRR in the UBHG2-10 showed a highest value ($4.29nmol\;cm^{-3}d^{-1}$) at the SMTZ. The 16S rRNA gene copy numbers of total Prokaryotes, mcrA, (methyl coenzyme M reductase subunit A), and dsrA (dissimilatory sulfite reductase subunit A) showed the peaks in the SMTZ at both sites, but the maximum mcrA gene copy number of the UBGH2-10 appeared below the SMTZ (9.8 mbsf). ANME-1 was a predominant ANME (Anaerobic MEthanotroph) group in both SMTZs of the UBGH2-3 and -10. However, The sequences of ANME-2 were detected only at 2.2 mbsf of the UBGH2-3 where high methane flux was observed because of massive amount of gas hydrate at shallow depth. And Desulfosarcina-Desulfococcus (DSS) that is associated with ANME-2 was detected in 2.2 mbsf of the UBHG2-3. Overall results demonstrate that ANME-1 and ANME-2 are considered as significant archaeal groups related to methane cycle in the subsurface sediment of the East Sea, and ANME-2/DSS consortia might be more responsible for methane oxidation in the methane seeping region than in non-seeping region.

Effects of Protein Level and Mangosteen Peel Pellets (Mago-pel) in Concentrate Diets on Rumen Fermentation and Milk Production in Lactating Dairy Crossbreds

  • Norrapoke, T.;Wanapat, M.;Wanapat, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.7
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    • pp.971-979
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    • 2012
  • Four, lactating dairy crossbreds ($50%{\times}50%$ Holstein Friesian${\times}$Native Zebu cattle) were randomly assigned according to a $2{\times}2$ factorial arrangement (two protein levels and two levels of mangosteen peel pellets (Mago-pel)) in a $4{\times}4$ Latin square design to receive four dietary treatments. All cows received concentrate at a proportion of 1 kg concentrate per 2 kg of milk yield, and urea-treated 5% rice straw (UTRS) was given ad libitum. It was found that total dry matter intakes, nutrient digestibility, ruminal pH and $NH_3$-N concentrations were not affected (p>0.05) by treatments. Concentrations of ruminal pH and $NH_3$-N were not affected by dietary treatments although the concentration of BUN varied significantly (p<0.05) between protein levels (p<0.05). The populations of rumen bacteria and fungal zoospores did not differ among treatments (p>0.05); however, the population of protozoa was decreased (p<0.05) when cows received Mago-pel supplementation. The composition of the population of bacteria, identified by real-time PCR technique, including total bacteria, methanogens, Fibrobacter succinogenes and Ruminococcus albus was similar (p>0.05) among dietary treatments (p>0.05); however, copy numbers of Ruminococcus flavefaciens was increased when protein level increased (p<0.05). Microbial protein synthesis, in terms of both quantity and efficiency, was enriched by Mago-pel supplementation. Milk yield was greatest in cows fed UTRS based diets with concentrate containing protein at 16% CP with Mago-pel, but were lowest without Mago-pel (p<0.05). In addition, protein level and supplementation of Mago-pel did not affect (p>0.05) milk composition except solids-not-fat which was higher in cows fed the diet with 19% CP. Therefore, feeding a concentrate containing 16% CP together with 300 g/hd/d Mago-pel supplementation results in changes in rumen fermentation and microbial population and improvements in milk production in lactating dairy crossbreds fed on UTRS.

Production of ʟ-Theanine Using Escherichia coli Whole-Cell Overexpressing γ-Glutamylmethylamide Synthetase with Baker's Yeast

  • Yang, Soo-Yeon;Han, Yeong-Hoon;Park, Ye-Lim;Park, Jun-Young;No, So-young;Jeong, Daham;Park, Saerom;Park, Hyung Yeon;Kim, Wooseong;Seo, Seung-Oh;Yang, Yung-Hun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.5
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    • pp.785-792
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    • 2020
  • ʟ-Theanine, found in green tea leaves has been shown to positively affect immunity and relaxation in humans. There have been many attempts to produce ʟ-theanine through enzymatic synthesis to overcome the limitations of traditional methods. Among the many genes coding for enzymes in the ʟ-theanine biosynthesis, glutamylmethylamide synthetase (GMAS) exhibits the greatest possibility of producing large amounts of production. Thus, GMAS from Methylovorus mays No. 9 was overexpressed in several strains including vectors with different copy numbers. BW25113(DE3) cells containing the pET24ma::gmas was selected for strains. The optimal temperature, pH, and metal ion concentration were 50℃, 7, and 5 mM MnCl2, respectively. Additionally, ATP was found to be an important factor for producing high concentration of ʟ-theanine so several strains were tested during the reaction for ATP regeneration. Baker's yeast was found to decrease the demand for ATP most effectively. Addition of potassium phosphate source was demonstrated by producing 4-fold higher ʟ-theanine. To enhance the conversion yield, GMAS was additionally overexpressed in the system. A maximum of 198 mM ʟ-theanine was produced with 16.5 mmol/l/h productivity. The whole-cell reaction involving GMAS has greatest potential for scale-up production of ʟ-theanine.

Effects of Tillage on Organic Matters and Microbial Communities in Organically Cultivated Corn Field Soils (유기농 옥수수밭에서 경운이 토양 유기물 함량 및 미생물군집에 미치는 영향)

  • Ahn, Dalrae;An, Nan-Hee;Kim, Da-Hye;Han, Byeong-Hak;You, Jaehong;Park, InCheol;Ahn, Jae-Hyung
    • Korean Journal of Environmental Agriculture
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    • v.39 no.1
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    • pp.65-74
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    • 2020
  • BACKGROUND: Soil carbon sequestration has been investigated for a long time because of its potential to mitigate the greenhouse effect. No- or reduced tillage, crop rotations, or cover crops have been investigated and practiced to sequester carbon in soils but the roles of soil biota, particularly microorganisms, have been mostly ignored although they affect the amount and stability of soil organic matters. METHODS AND RESULTS: In this study we analyzed the organic matter and microbial community in organically cultivated corn field soils where no-tillage (NT) or conventional tillage (CT) had been practiced for about three years. The amounts of organic matter and recalcitrant carbon pool were 18.3 g/kg dry soil and 4.1 g C/kg dry soil, respectively in NT soils, while they were 12.4 and 2.5, respectively in CT soils. The amounts of RNA and DNA, and the copy numbers of bacterial 16S rRNA genes and fungal ITS sequences were higher in NT soils than in CT soils. No-tillage treatment increased the diversities of soil bacterial and fungal communities and clearly shifted the bacterial and fungal community structures. In NT soils the relative abundances of bacterial phyla known as copiotrophs, Betaproteobacteria and Bacteroidetes, increased while those known as oligotrophs, Acidobacteria and Verrucomicrobia, decreased compared to CT soils. The relative abundance of a fungal phylum, Glomeromycota, whose members are known as arbuscular mycorrhizal fungi, was about two time higher in NT soils than in CT soils, suggesting that the higher amount of organic matter in NT soils is related to its abundance. CONCLUSION: This study shows that no-tillage treatment greatly affects soil microbial abundance and community structure, which may affect the amount and stability of soil organic matter.

The Uneven Distribution of Mating Type Genes in Natural and Cultivated Truffle Orchards Contributes to the Fructification of Tuber indicum

  • Li, Qiang;Fu, Yu;Sun, Qun;Sourzat, Pierre;Yang, Mei;Liu, Chengyi;Tan, Hao;Ye, Lei;Zou, Jie;Wu, Chenguang;Zhang, Bo;Li, Xiaolin
    • Mycobiology
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    • v.46 no.1
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    • pp.64-71
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    • 2018
  • The aim of this study was to investigate the pattern of distribution of mating type (MAT) genes of Tuber indicum in ectomycorhizosphere soils from natural T. indicum-producing areas and cultivated truffle orchards and ascocarp samples from different regions. Quantitative real-time PCR and multiplex PCR were used to weight the copy numbers of MAT1-1-1 and MAT1-2-1 in natural truffle soils and cultivated orchard soils. The effect of limestone on the pattern of truffle MAT genes and the correlation between soil properties and the proportion of MAT genes were also assessed. These results indicated that an uneven and nonrandom distribution of MAT genes was common in truffle-producing areas, cultivated truffle orchards, and ascocarps gleba. The competition between the two mating type genes and the expansion of unbalanced distribution was found to be closely related to truffle fructification. Limestone treatments failed to alter the proportion of the two mating type genes in the soil. The content of available phosphorus in soil was significantly correlated with the value of MAT1-1-1/MAT1-2-1 in cultivated and natural ectomycorhizosphere soils. The application of real-time quantitative PCR can provide reference for monitoring the dynamic changes of mating type genes in soil. This study investigates the distributional pattern of T. indicum MAT genes in the ectomycorhizosphere soil and ascocarp gleba from different regions, which may provide a foundation for the cultivation of T. indicum.

Evaluation of liquid and powdered forms of polyclonal antibody preparation against Streptococcus bovis and Fusobacterium necrophorum in cattle adapted or not adapted to highly fermentable carbohydrate diets

  • Cassiano, Eduardo Cuellar Orlandi;Perna, Flavio Junior;Barros, Tarley Araujo;Marino, Carolina Tobias;Pacheco, Rodrigo Dias Lauritano;Ferreira, Fernanda Altieri;Millen, Danilo Domingues;Martins, Mauricio Furlan;Pugine, Silvana Marina Piccoli;de Melo, Mariza Pires;Beauchemin, Karen Ann;Meyer, Paula Marques;Arrigoni, Mario de Beni;Rodrigues, Paulo Henrique Mazza
    • Animal Bioscience
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    • v.34 no.1
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    • pp.74-84
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    • 2021
  • Objective: Feed additives that modify rumen fermentation can be used to prevent metabolic disturbances such as acidosis and optimize beef cattle production. The study evaluated the effects of liquid and powdered forms of polyclonal antibody preparation (PAP) against Streptococcus bovis and Fusobacterium necrophorum on rumen fermentation parameters in ruminally cannulated non-lactating dairy cows that were adapted or unadapted to a high concentrate diet. Methods: A double 3×3 Latin square design was used with three PAP treatments (control, powdered, and liquid PAP) and two adaptation protocols (adapted, unadapted; applied to the square). Adapted animals were transitioned for 2 weeks from an all-forage to an 80% concentrate diet, while unadapted animals were switched abruptly. Results: Interactions between sampling time and adaptation were observed; 12 h after feeding, the adapted group had lower ruminal pH and greater total short chain fatty acid concentrations than the unadapted group, while the opposite was observed after 24 h. Acetate:propionate ratio, molar proportion of butyrate and ammonia nitrogen concentration were generally greater in adapted than unadapted cattle up to 36 h after feeding. Adaptation promoted 3.5 times the number of Entodinium protozoa but copy numbers of Streptococcus bovis and Fibrobacter succinogens genes in rumen fluid were not affected. However, neither liquid nor powdered forms of PAP altered rumen acidosis variables in adapted or unadapted animals. Conclusion: Adaptation of cattle to highly fermentable carbohydrate diets promoted a more stable ruminal environment, but PAP was not effective in this study in which no animal experienced acute or sub-acute rumen acidosis.

Expansion of Personal Identification to Vitalize the Use of Unmanned Civil Service Issuing Machines for Gongju-si (공주시 무인민원발급기 이용 활성화를 위한 신분 확인 확대 방안)

  • Kim, Eui Seok;Jung, Hanmin
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2021.10a
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    • pp.156-158
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    • 2021
  • An initial local government center received direct asking from citizens, wrote the documents by hand, and issued them. Later the appearance of computers, printers, and faxes made it possible to handle civil complaints through copying. Since 2010, as the e-government has started, entering civil service requests into computers after identification by themselves and printing the results has become established. In addition, with the introduction of unmanned administrative services, civil services become available in various places, which causes the increased satisfaction of citizens. One of the services is an unmanned civil service issuing machine, and more than 20 kinds of documents, including a copy of resident registration, can be issued directly. Of course, additional help may be needed if citizens are not familiar with the latest technologies. However, the unmanned civil service issuing machine is a highly satisfactory way in that it is less time-limited as citizens can visit a community center or city hall after the end of work. However, the main problem is that resident registration numbers and fingerprint recognition are required depending on the type of issuance, which increases the inconvenience of handling citizen complaints and is shunned. To solve this problem, we aim to study a method for improving the fingerprint recognition method of unmanned civil service issuing machines by expanding personal identification.

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Detection of Campylobacter jejuni in food and poultry visors using immunomagnetic separation and microtitre hybridization

  • Simard, Ronald-E.
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.71-73
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    • 2000
  • Campylobacter jejuni is most frequently identified cause of cause of acute diarrhoeal infections in developeed countries, exceeding rates of illness caused by both salmonella and shigilla(Skirrow, 1990 ; Lior 1994). Previous studies on campylobacter jejuni contamination of commercial broiler carcasses in u.s.(Stern, 1992). Most cases of the disease result from indirect transmission of Campylobactor from animals via milk, water and meat. In addition to Campylobactor jejuni. the closely relates species Campylobactor coli and Campylobactor lari have also been implicated as agents of gastroenteritis in humans. Campylobactor coli represented only approximately 3% of the Campylobactor isolates from patients with Campylobactor enteritis(Griffiths and Park, 1990) whereas Campylobactor coli is mainly isolated from pork(Lmmerding et al., 1988). Campylobactor jejuni has also been isolated from cases of bacteremia, appendicitis and, recently, has been associated with Guillai-Barre syndrome(Allos and Blaser, 1994; von Wulffen et al., 1994; Phillips, 1995). Studies in volunteers indicated that the infectious dose for Campylobactor jejuni is low(about 500 organisms)(Robinson, 1981). The methods traditionally used to detect Campylobactor ssp. in food require at least two days of incubation in an enrichment broth followed by plating and two days of incubation on complex culture media containing many antibiotics(Goossens and Butzler, 1992). Finnaly, several biochemical tests must be done to confirm the indentification at the species level. Therfore, sensitive and specific methods for the detection of small numbers of Campylobactor cells in food are needed. Polymerase chain reaction(PCR) assays targeting specific DNA sequences have been developed for the detection of Campylobactor(Giesendorf and Quint, 1995; Hemandex et al., 1995; Winter and Slavidk, 1995). In most cases, a short enrichment step is needed to enhance the sensitivity of the assay prior to detection by PCR as the number of bacteria in the food products is low in comparison with those found in dinical samples, and because the complex composition of food matrices can hinder the PCR and lower its sensitivity. However, these PCR systems are technically demanding to carry out and cumbersome when processing a large number of samples simutaneously. In this paper, an immunomagnetic method to concentrate Campylobactor cells present in food or clinical samples after an enrichment step is described. To detect specifically the thermophilic Campylobactor. a monoclonal antibody was adsorbed on the surface of the magnetic beads which react against a major porin of 45kDa present on the surface of the cells(Huyer et al., 1986). After this partial purification and concentration step, detection of bound cells was achieved using a simple, inexpensive microtitre plate-based hybridization system. We examined two alternative detection systems, one specific for thermophilic Campylobactor based on the detection of 23S rRNA using an immobilized DNA probe. The second system is less specific but more sensitive because of the high copy number of the rRNA present in bacterial cell($10^3-10^4$). By using specific immunomagnetic beads against thermophilic Campylobactor, it was possible to concentrate these cells from a heterogeneous media and obtain highly specific hybridization reactions with good sensitivity. There are several advantages in using microtitre plates instead of filter membranes or other matrices for hybridization techniques. Microtitre plates are much easier to handle than filter membranes during the adsorption, washing, hybridization and detection steps, and their use faciilitates the simultanuous analysis of multiple sample. Here we report on the use of a very simple detection procedure based on a monoclonal anti-RNA-DNA hybrid antibody(Fliss et al., 1999) for detection of the RNA-DNA hybrids formed in the wells.

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