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Identification of Lactic Acid Bacteria in Galchi- and Myeolchi-Jeotgal by 16S rRNA Gene Sequencing, MALDI-TOF Mass Spectrometry, and PCR-DGGE

  • Lee, Yoonju;Cho, Youngjae;Kim, Eiseul;Kim, Hyun-Joong;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1112-1121
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    • 2018
  • Jeotgal is a Korean traditional fermented seafood with a high concentration of salt. In this study, we isolated lactic acid bacteria (LAB) from galchi (Trichiurus lepturus, hairtail) and myeolchi (Engraulis japonicas, anchovy) jeotgal on MRS agar and MRS agar containing 5% NaCl (MRS agar+5% NaCl), and identified them by using 16S rRNA gene sequencing and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) as culture-dependent methods. We also performed polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) as a culture-independent method to identify bacterial communities. Five samples of galchi-jeotgal and seven samples of myeolchi-jeotgal were collected from different regions in Korea. A total of 327 and 395 colonies were isolated from the galchi- and myeolchi-jeotgal samples, respectively. 16S rRNA gene sequencing and MALDI-TOF MS revealed that the genus Pediococcus was predominant on MRS agar, and Tetragenococcus halophilus on MRS agar+5% NaCl. PCR-DGGE revealed that T. halophilus, Tetragenococcus muriaticus, and Lactobacillus sakei were predominant in both types of jeotgal. T. halophilus was detected in all samples. Even though the same species were identified by both culture-dependent and -independent methods, many species identified by the culture-dependent methods were not in the bacterial list identified by the culture-independent methods. The distribution of bacteria in galchi-jeotgal was more diverse than in myeolchi-jeotgal. The diverse LAB in galchi- and myeolchi-jeotgals can be further studied as candidates for starter cultures to produce fermented foods.

Extended latex proteome analysis deciphers additional roles of the lettuce laticifer

  • Cho, Won-Kyong;Chen, Xiong-Yan;Rim, Yeong-Gil;Chu, Hyo-Sub;Jo, Yeon-Hwa;Kim, Su-Wha;Park, Zee-Yong;Kim, Jae-Yean
    • Plant Biotechnology Reports
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    • v.4 no.4
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    • pp.311-319
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    • 2010
  • Lettuce is an economically important leafy vegetable that accumulates a milk-like sap called latex in the laticifer. Previously, we conducted a large-scale lettuce latex proteomic analysis. However, the identified proteins were obtained only from lettuce ESTs and proteins deposited in NCBI databases. To extend the number of known latex proteins, we carried out an analysis identifying 302 additional proteins that were matched to the NCBI non-redundant protein database. Interestingly, the newly identified proteins were not recovered from lettuce EST and protein databases, indicating the usefulness of this hetero system in MudPIT analysis. Gene ontology studies revealed that the newly identified latex proteins are involved in many processes, including many metabolic pathways, binding functions, stress responses, developmental processes, protein metabolism, transport and signal transduction. Application of the non-redundant plant protein database led to the identification of an increased number of latex proteins. These newly identified latex proteins provide a rich source of information for laticifer research.

Analysis of Uterine Bacteria to Increase Reproductive Efficiency in Hanwoo(Korean Native Cattle) (한우의 번식 효율 향상을 위한 자궁 내 세균 분석)

  • Park, Joung-Jun;Yoo, Han-Jun;Cho, Young-Jae;Choi, Hye-Won;Yoon, Pil-Sang;Lee, Seon-Goo;Jung, Bae-Dong;Hahn, Tae-Wook
    • Journal of Embryo Transfer
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    • v.28 no.1
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    • pp.49-55
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    • 2013
  • The objective of this study was to evaluate several types of uterine bacteria in Hanwoo. uterine bacteria from randomly selected 5 uterus was collected by flushing methods into a sterilized 1.5 ml centrifuge tube and was inoculated onto MacConkey agar and blood agar, respectively. After being incubated for 5% $CO_2$, aerobic or anaerobic condition at $37^{\circ}C$ during 48h, bacterial colonies were selected and re-inoculated onto blood agar plates. Re-cultured colonies were identified by Gram staining and finally identified using Vitek system. The identified bacteria were Staphylococcus lentus, Staphylococcus sciuri, Staphylococcus vitulinus, Staphylococcus warneri of Gram (+) and Rhizobium radiobacter, Sphingomonas paucimobilis of Gram (-) bacteria. Although, pathogenicity of identified bacteria was unclear, the bacteria can have an effect on the uterine microenvironment. Therefore, repetitive research will be required to determine the effects of bacteria in cattle exposed to a various environment.

Identification of Flavor Components in Tomato Fruit (토마토 향기성분의 동정)

  • Sohn, Tae-Hwa;Cheon, Sung-Ho;Choi, Sang-Won;Moon, Kwang-Deok
    • Applied Biological Chemistry
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    • v.31 no.3
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    • pp.292-297
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    • 1988
  • The volatiles of whole and homogenated tomato fruits collected by the headspace trapping method using Tenax GC and the simultaneous steam distillation method were identified by GC and GC-MS. Among over 100 GC peaks, 10 alcohols, 6 aldehydes, 4 ketones, 3 esters, 1 phenol and 1 acid were identified from whole tomato fruits, whereas 12 alcohols, 6 aldehydes, 5 ketones, 5 esters, 2 phenols, 1 hydrocarbon and 1 acid were identified from homogenated tomato fruits. By simultaneous steam distillation-extraction, 19 alcohols, 13 hydrocarbons, 9 esters, 9 ketones, 8 aldehydes, 2 phenols, 2 lactones,2 furans, 1 acids and 2 others were identified among over 300 peaks.

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New Frontiers of Knowledge on Nepalese Plant Science

  • Shrestha, Gyan L.;Shrestha, Bhushan
    • Korean Journal of Weed Science
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    • v.14 no.1
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    • pp.71-80
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    • 1994
  • Nepal, with its unique geographical and ecological features due to its abrupt rise in altitude, plays significant role in biological evolution. Existence of numerous wild relatives of the present-day cultivated agricultural crop plants in this small Himalayan nation may serve as a potential source of several yet unidentified desirable genes that are needed for future incorporation in the improvement of cultivated crop plants. This report includes 82 different wild relatives of 41 genera under 19 families of 37 agricultural crops of Nepal(Table 1). It serves as the sample of the glossary of these wild relatives of crop plants in Nepal. Under food grain crop plants of gramineae, leguminoceae and polygonaceae families, 16 different wild species namely wild rices(7 species), wild relatives of wheat plant(3 species), wild arhar(3 species), wild fingermillets(1 species) and wild buckwheat(2 species) have been identified in different parts of the country. Similarly, under vegetable crop plants of Araceae, Amaranthaceae, Crucifereae, Cucurbitaceae, Dioscoreaceae, Labiteae, Leguminosae, Liliaceae, Malvaceae, Polygonaceae, Solanaceae and Umbellifereae, 37 different wild species-wild colocasia(1 species), wild amaranths(3 species), wild leafy vegetables(2 species), wild gourds(3 species), wild cucumber(1 species), wild yams(4 species), wild mints(3 species), wild fenugreeks(4 species), wild pea(1 species), wild beans(3 species), wild garlics(2 species), wild spinach(3 species), wild lady's finger(1 species), wild spinach(3 species), wild eggplants(2 species) and wild carrot(one species) have also been identified. In case of wild relatives of cultivated orchard plants, 11 different wild species namely wild mango(one species), wild banana(one species), wild strawberry(one species), wild pear(one species), wild cherries(2 species), wild apple(one species) and wild grapes(3 species) have been identified, Among 19 different wild species of economic crop plants, five wild species of sugarcane, one species of wild sunhemp, two wild relatives of cotton, three wild relatives of rose, two wild species of tobacco, four wild species of turmeric and two wild species of tea have also been identified. This report includes only sample of the total wild species of the present-day cultivated agricultural crop plants. Further exploration on this economic botany will help the country in cataloging the wild relatives of cultivated crop plants and their future use in crop improvement.

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Identification of Copper and Cadmium Induced Genes in Alfalfa Leaves through Annealing Control Primer Based Approach

  • Lee, Ki-Won;Rahman, Md. Atikur;Zada, Muhammad;Lee, Dong-Gi;Kim, Ki-Yong;Hwang, Tae Young;Ji, Hee Jung;Lee, Sang-Hoon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.35 no.3
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    • pp.264-268
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    • 2015
  • The present research investigated copper and cadmium stress-induced differentially expressed genes (DEGs) using annealing control primers (ACP) with the differential display reverse transcription polymerase chain reaction technique in alfalfa (Medicago sativa L. cv. Vernal) leaves. Alfalfa leaves were subjected to $250{\mu}M$ of copper and cadmium treatment for a period of 6 h. A total of 120 ACPs was used. During copper and cadmium treatment, 6 DEGs were found to be up or down regulated. During copper stress treatment, 1 DEG was up-regulated, and 3 novel genes were discovered. Similarly, during cadmium stress treatment, 1 DEG was up-regulated and 5 novel genes were identified. Among all 6 DEGs, DEG-4 was identified as the gene for trans-2,3-enoyl-CoA reductase, DEG-5 was identified as the gene for senescence-associated protein DIN1 and DEG-6 was identified for caffeic acid O-methyltransferase. All the up-regulated genes may play a role in copper and cadmium stress tolerance in alfalfa.

Analysis of Bovine Seminal Plasma Proteins from Korean Native Cattle, Hanwoo, and Korean Native Brindle Cattle

  • Lee, Su-Rok;Kim, Eun-Sung;Kim, Sung-Woo;Kim, Hyeong-Chul;Shim, Kwan-Seob;Kim, Jong-Gug
    • Reproductive and Developmental Biology
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    • v.36 no.2
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    • pp.121-131
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    • 2012
  • After spermatogenesis, spermatozoa come in contact with fluids in the epididymis where they mature. During ejaculation, spermatozoa are mixed with secretions from prostate gland, vesicular glands, and bulbourethral glands. During natural mating, seminal plasma is deposited in the female reproductive tract eliciting various physiological and immunological responses. With the advances in proteomics, the components of seminal plasma have been identified and the information may be valuable in identifying markers for fertility. Components of seminal plasma that affect fertility have been discovered and the mechanism of action of these factors has been determined. The objective of this study was to determine the specific seminal plasma proteins from Korean native cattle, Hanwoo, and Korean native brindle cattle (KNBC) with the long term goal of improving fertilization rate. After SDS-PAGE and 2-dimensional gel electrophoresis, proteins were identified by Q-ToF analysis. They include plasma serine protease inhibitor precursor and platelet-activating factor acetylhydrolase after SDS-PAGE. Number and density of the spots in 2-dimensional gels were higher in KNBC than Hanwoo. Proteins identified from the paired spots of both breeds include chain A, bull seminal plasma PDC-109 Fibronectin Type II module, BSP-30 kDa precursor, and Spermadhesin Z13 or its precursor. Interestingly, some proteins were identified from multiple spots. The functional differences of these diverse forms of the proteins may require further studies. With their previously reported roles in sperm capacitation by these proteins, the studies on the mechanism of action, ligand interaction and the variation in the genome may help improving fertility in cattle.

Identification of Single Nucleotide Polymorphisms in PRNP Gene of Korean Native Goats

  • Hoque, Md. Rashedul;Yu, Seong-Lan;Yeon, Seong-Heum;Lee, Jun-Heon
    • Journal of Animal Science and Technology
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    • v.51 no.6
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    • pp.453-458
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    • 2009
  • Prion protein (PRNP) is known to be a causative protein for transmissible spongiform encephalopathy (TSE), a disease occurring in human and animals. Previous results indicate that the genetic variability can affect the resistance and susceptibility of goat scrapie and can give the guideline for reducing the risk of this disease. Until now, 35 single nucleotide polymorphisms (SNPs) were identified in goat PRNP gene from many countries such as Great Britain, Italy, United States of America and Asian countries etc. In this study, SNPs in PRNP gene have been investigated to research the PRNP variations and their possible TSE risks in 60 Korean native goats. Based on the sequencing results, we identified four SNPs and three of those polymorphisms (G126A, C414T and C718T) were synonymous and the A428G polymorphism was non-synonymous which changes the amino acid histidine to arginine. Previously, all of these four SNPs were identified in Asian native goats. Specifically, five polymorphisms were identified in Asian native goats and two of them (G126A and C414T) were silent mutations, and the other SNPs (T304G, A428G and T718C) caused amino acid changes (W102G, H143R and S240P). Comparing with SNP results from other breeds, this study is an initial step to understand resistance and susceptibility of this disease in Korean native goats.

Identification of Coliform Bacteria Isolated from Nangmyun-Broth in Korea and Psychrotrophic Character (냉면육수에서 분리한 대장균군의 동정 및 저온 증식성)

  • O, Myeong-Hwan;Kim, Mi-Yeong;Lee, Jin-Yeong
    • The Korean Journal of Food And Nutrition
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    • v.7 no.3
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    • pp.203-212
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    • 1994
  • The nangmyun is a Korean iced noodle made by putting buckwheat-noodles into cold broth. Seven samples of nangmyun-broths were collected from Korean restaurants in Bucheon during July 1994. The contamination levels of nangmyun-broths by coliform bacteria were determined, and then the 40 colonies of coliform bacteria, isolated randomly from 4 samples of nangmyun-broths, were identified at genus or species level with the additional test for psychrotrophic character The coliform counts in nangmyun-broths were 6.0x102-6.5$\times$104/ml(average 2.3$\times$104%). Among the 40 strains of isolates, 27 strains(67.5%) were identified as the genus Klebsiella, 9 strains(22.5%) as the genus Enterobacter, 2 strains(5.0%) as the genus Citrobacter and 2 strains (5.0%) as the genus Escherichia. Among 27 strains of Klebsiella, 11 strains (40.8%) were identified as K planticola, 4 strains(14.8%) as K. pneumoniae, 2 strains(7.4%) as K. ozaenae and 2 strains(7.4%) as K. terrigena, but 8 strains(29.6%) of a typic\ulcorner1 Klebgiella could not be Identified at species level. All the 40 strains of coliform bacteria were psychrotrophs showing slow growth at 1$0^{\circ}C$, and 18 strains(45%) grew at 5$^{\circ}C$. It was thought to be a good basic data In describing the reason for too high coliform counts in nangmyun-broths that all coliform bacteria tested In this study were psychrotrophs.

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Analyses of the Volatile Flavor Composition of Essential Oils from Chrysanthemum zawadskii var. latilobum Kitamura and Aster yomena Makino (구절초와 쑥부쟁이 정유의 휘발성 향기성분 분석)

  • Choi, Hyang-Sook
    • The Korean Journal of Food And Nutrition
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    • v.31 no.3
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    • pp.378-387
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    • 2018
  • This study investigated the volatile flavor composition of essential oils from Chrysanthemum zawadskii var. latilobum Kitamura and Aster yomena Makino. The essential oils obtained by the hydrodistillation extraction method from the aerial parts of the plants were analyzed by gas chromatography (GC) and GC-mass spectrometry (GC-MS). One hundred (95.04%) volatile flavor compounds were identified in the essential oil from the C. zawadskii var. latilobum Kitamura. The major compounds were valencene (10.82%), ${\delta}$-cadinol (9.77%), hexadecanoic acid (8.70%), 2-methyl-4-(2,6,6-trimethylcyclohex-1-enyl) but-2-en-1-ol (3.67%), and 2-(2,4-hexadiynylidene)-1,6-dioxaspiro[4,4]non-3-ene (3.57%). Ninety-eight (93.83%) volatile flavor compounds were identified in the essential oil from the Aster yomena Makino. The major compounds were and 3-eicosyne (13.61%), 9,10,12-octadecatrienoic acid (7.8%), ${\alpha}$-caryophyllene alcohol (6.83%), 9-octadecynoic acid (6.03%), and ${\alpha}$-caryophyllene (5.74%). Although the two plants are apparently very similar, the chemical composition of the essential oils was significantly different in quality and quantity. In the case of C. zawadskii var. latilobum Kitamura, the sesquiterpene, valencene was found to be 10.82%, but it was not identified in A. yomena Makino. ${\delta}$-Cadinol appeared higher in C. zawadskii var. latilobum Kitamura than in A. yomena Makino. A clear characteristic of A. yomena Makino essential oil is that it has a high content of caryophyllene derivatives. The ${\alpha}$-caryophyllene alcohol contained in A. yomena Makino was relatively high at 6.83%, although the compound was not identified in C. zawadskii var. latilobum Kitamura. Also ${\alpha}$-caryophyllene was shown to be higher in A. yomena Makino than in C. zawadskii var. latilobum Kitamura.