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In vitro micropropagation of M.26 (Malus pumila Mill) apple rootstock and assessment of the genetic diversity of proliferated plantlets using simple sequence repeat markers (사과 대목 M.26 (Malus pumila Mill)의 기내 대량번식 및 simple sequence repeat 마커를 이용한 증식된 식물체의 유전적 다양성 평가)

  • Cho, Kang Hee;Han, Bong Hee;Han, Jeom Hwa;Park, Seo Jun;Kim, Se Hee;Lee, Han Chan;Kim, Mi Young;Kim, Myung-Su
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.382-391
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    • 2018
  • The objective of this study was to determine the most effective medium condition of shoot proliferation and root formation for the efficient in vitro micropropagation of M.26 (Malus pumila Mill). Simple sequence repeat (SSR) markers were used to analyze the genetic diversity of micro-propagated and greenhouse grown M.26. Shoot proliferation was carried out in MS (Murashige and Skoog) containing benzyladenin (BA, $0.5{\sim}5.0mg{\cdot}L^{-1}$) and thidiazuron (TDZ, $0.01{\sim}0.1mg{\cdot}L^{-1}$). The highest number of shoots (10.67 shoots per explant) was induced by adding BA at a concentration $1.0mg{\cdot}L^{-1}$. TDZ treatments caused higher hyperhydricity rate in cultured explants than in BA treatments. There was no significant effect of both BA and auxin on shoot proliferation, and the optimum proliferation medium for M.26 was MS medium containing $1.0mg{\cdot}L^{-1}$ BA. To find a suitable medium composition for shoot rooting, we tested different concentrations indole-3-butyric acid (IBA) and ${\alpha}$-naphthaleneacetic acid ($0.5{\sim}5.0mg{\cdot}L^{-1}$), MS medium (1/4-1), sucrose ($0{\sim}30g{\cdot}L^{-1}$). The shoots showed good rooting on half-strength MS medium containing $1.0mg{\cdot}L^{-1}$ IBA and $15-20g{\cdot}L^{-1}$ sucrose. The rooting rate (100%), number of roots (10.45 ~ 13.60 roots per explant), root length (7.41 ~ 8.33 cm), and shoot length (4.93 ~ 5.38 cm) were good on this medium. Fifteen SSR primers were detected in a total of 30 alleles in 20 micro-propagated plantlets, all SSR profiles from micro-propagated plantlets were monomorphic and similar to greenhouse grown control plantlet M.26 plant. The results indicated that M.26 micro-propagated plantlets were genetically stable.

In vitro Multiplication of Hosta Tratt. Species Native to Korea by Shoot-tip Culture (경정배양에 의한 한국 자생 비비추속 식물의 기내증식)

  • Choi, Han;Yang, Jong Cheol;Ryu, Sun Hee;Yoon, Sae Mi;Kim, Sang Yong;Lee, Seung Youn
    • Korean Journal of Plant Resources
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    • v.32 no.1
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    • pp.53-62
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    • 2019
  • The purpose of this study was to establish the in vitro propagation system by shoot tip culture of six Hosta species native to Korea (Hosta capitata (Koidz.) Nakai, H. clausa Nakai, H. jonesii M.G.Chung, H. minor (Baker) Nakai, H. venusta F.Maek., and H. yingeri S.B.Jones) for mass proliferation and a new cultivar development. The shoot tips of each Hosta species were cultured on MS medium containing eight combinations of 0.5, 1.0, 2.0, 4.0 mg/L BA with 0.1 mg/L NAA, 0.1, 0.5, 1.0, 2.0 mg/L TDZ with 0.1 mg/L NAA, and without any PGRs (control). They were investigated on callus, somatic embryo, crown bud, differentiation and growth of shoot and root, total fresh weight after 8 weeks of culture. In all six Hosta species, callus and somatic embryo induction rate and multiple shooting rate of the PGRs treatment group were higher than that of the control group. The highest number of differentiated shoots were obtained on medium supplemented with 2.0 ㎎/L TDZ in H. capitata (5.4), 1.0 mg/L TDZ in H. clausa and H. jonesii (3.3 and 5.8, respectively), 0.5 mg/L BA in H. minor (11.1), 1.0 mg/L BA and 0.1 mg/L TDZ in H. venusta (8.1), and 0.5 mg/L TDZ in H. yingeri (9.8). In somatic embryo formation, the PGRs treatment group of H. jonesii and H. yingeri were more effective than the control group, and the effects were relatively less in H. capitata, H. clausa Nakai, H. minor, H. venusta. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. H. clausa showed no significant effect on callus and shoot differentiation regardless of the type and concentration of cytokinin, but slightly increased in formation of crown bud in TDZ.

Mycelial and cultural characteristics of Pleurotus ostreatus 'Baekseon', a novel white cultivar for bottle culture (병재배용백색느타리신품종 『백선』의 균사배양및생육특성)

  • Choi, Jong In;Lee, Yun Hae;Gwon, Hee Min;Jeon, Dae Hoon;Lee, Yong Seon;Lee, Young Sun
    • Journal of Mushroom
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    • v.17 no.3
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    • pp.113-118
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    • 2019
  • Oyster mushrooms are an economically important crop, accounting for 35% of the total mushroom production in Korea. In this study, we developed a new cultivar of Pleurotus ostreatus, known as 'Baekseon,' which is characterized by a white pileus with a white stipe. It was bred by mating monokaryons isolated from white mutant oyster mushrooms that were naturally generated from 'Gonji-7ho' and 'Wonhyeong-1ho' at the Mushroom Research Institute, GARES, Korea in 2018. The optimum temperature for mycelial growth on potato dextrose agar medium was approximately $28-31^{\circ}C$, and the optimum temperatures for primordia formation and growth of fruit bodies on sawdust media were $22^{\circ}C$ and $20^{\circ}C$, respectively. The time required for the bottle-cultured mushrooms to complete spawn running, primordia formation, and growth of fruit bodies was 30 days, 4 days, and 4 days, respectively. The fruit bodies were bundle-shaped, the pilei were round type and white, and the stipes were white. The stipes were slender and longer than those of the control ('Miso'). In the productivity test, the yield per bottle was 185 g/1100 mL, which was 45% greater than that of the control ('Miso'). In the farm test, the yield per bottle for Farm A (Pyeongtaek) and Farm B (Yeoju) was 184 g/1100 mL and 178 g/850 mL, respectively. With regard to the physical properties of fruit bodies, the springiness, cohesiveness, gumminess, and brittleness of stipe tissue were 80%, 57%, 720 g, and 57 kg, respectively. These values were lower than those of the control ('Miso'). To test the shelf life, the fruit bodies were wrapped with antifogging film and stored at $4^{\circ}C$ for 28 days and then at room temperature for 4 days; such conditions were sufficient for maintaining edibility.

Effect of Prostaglandin F2 Alpha on E-cadherin, N-cadherin and Cell Adhesion in Ovarian Luteal Theca Cells (난소의 황체협막세포에서 E-cadherin, N-cadherin과 세포부착에 미치는 Prostaglandin F2 Alpha의 영향)

  • Lee, Sang-Hee;Jung, Bae Dong;Lee, Seunghyung
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.360-369
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    • 2019
  • Cadherins are essential transmembrane proteins that promote cell-cell adhesion and maintain the corpus luteum structure in the ovary. This study examined the influence of prostaglandin F2 alpha ($PGF2{\alpha}$) on E-cadherin, N-cadherin, and adhesion in luteal theca cells (LTCs). The luteal cells were isolated from the mid-phase corpus luteum, and the LTCs were cultured separately from the luteal heterogeneous cells according to the morphology of the mesenchymal cells and to determine if steroidogenic and endothelial cells of LTCs, 3beta-hydroxysteroid dehydrogenase ($3{\beta}$-HSD), and vascular endothelial growth factor receptor 2 (VEGFR2) mRNA were used. The LTCs were then incubated in the culture medium supplemented with 0.01, 0.1, and 1.0 mM $PGF2{\alpha}$ for 24 h, and the E-cadherin and N-cadherin proteins in the LTCs were detected by confocal laser scanning microscopy. The results revealed $3{\beta}$-HSD mRNA expression in the LTC but no VEGF2R mRNA expression. The E-cadherin and N-cadherin proteins of the LTCs were damaged in the 0.01, 0.1, and 1.0 mM $PGF2{\alpha}$ treatment groups, and the expression of the N-cadherin protein was reduced significantly in 0.01 mM $PGF2{\alpha}$ compared to the 0 mM $PGF2{\alpha}$ treatment groups (P<0.05). In addition, the number of attached LTCs were significantly lower in the 0.01 mM $PGF2{\alpha}$ treatment group than in the 0 mM $PGF2{\alpha}$ treatment group (P<0.05). In conclusion, $PGF2{\alpha}$ affected the disruption of cadherin proteins and cell adhesion in LTCs. These results may help better understand the cadherin and adhesion mechanism during corpus luteum regression in the ovary.

Role of Dual Oxidase 2 in Reactive Oxygen Species Production Induced by Airborne Particulate Matter PM10 in Human Epidermal Keratinocytes (인간 표피 각질형성세포에서 대기 미립자 물질 PM10에 의해 유도되는 반응성 산소종의 생성에서 Dual oxidase 2의 역할)

  • Seok, Jin Kyung;Choi, Min A;Ha, Jae Won;Boo, Yong Chool
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.1
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    • pp.57-67
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    • 2019
  • Particulate matters with a diameter of < $10{\mu}m$ (PM10) exert oxidative stress and inflammatory events in various organs. The purpose of this study was to examine the molecular mechanism of reactive oxygen species (ROS) production induced by PM10 in the human epidermal keratinocytes (HEKs). When cultured HEKs were exposed to PM10, ROS production was induced and it was inhibited by apocynin, an antioxidant. The mRNA expression of NADPH oxidase (NOX) family was analyzed in order to examine their role in PM10-induced ROS production. PM10 increased the mRNA expression of NOX1, NOX2, dual oxidase (DUOX) 1 and DUOX2. HEKs expressed DUOX1 and DUOX2 at higher levels compared to other NOXs. The mRNA expression of dual oxidase maturation factors, DUOXA1 and DUOXA2, was also increased by PM10. We examined whether these calcium-dependent enzymes, DUOX1 and DUOX2, mediate the PM10-induced ROS production. A selective intracellular calcium chelator, BAPTA-AM, attenuated ROS production induced by PM10 or calcium ionophore A23187. The small intereference RNA (siRNA)-mediated down-regulation of DUOX2, but not DUOX1, attenuated the ROS production induced by PM10. PM10 increased the expression of inflammatory cytokines such as interleukin $(IL)-1{\beta}$, IL-6, IL-8 and interferon $(IFN)-{\gamma}$. SiRNA-mediated down-regulation of DUOX2 suppressed the PM10-induced expression of $IFN-{\gamma}$ but not other cytokines. This study suggests that DUOX2 plays a crucial role in ROS production and inflammatory response in PM10-exposed keratinocytes.

Establishment of tissue culture and acclimatization method for in vitro mass propagation of Echeveria laui and Echeveria elegans (에케베리아 라우이(Echeveria laui)와 엘레강스(Echeveria elegans)의 대량증식을 위한 조직배양 및 순화 조건 확립)

  • Kim, Youn Hee;Lee, Gee Young;Kim, Hye Hyeong;Lee, Jae Hong;Jung, Jae Hong;Lee, Sang Deok
    • Journal of Plant Biotechnology
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    • v.46 no.1
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    • pp.22-31
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    • 2019
  • The objective of this study was to investigate the suitable parts for callus induction and optimal concentrations of growth regulators contained in the medium affecting shooting and rooting Echeveria laui and Echeveria elegans for in vitro mass production. To determine the suitable plant parts for callus induction, the leaves were divided into upper, medium and bottom parts and cultured on MS medium at different concentrations with $0{\sim}2mgL^{-1}\;NAA$ and $0{\sim}4 mgL^{-1}BA$. The upper and middle parts of leaves both showed 100% callus formation rate with $NAA\;1\;mgL^{-1}$ and $BA\;1\;mgL^{-1}$ treatment in E. laui. The middle parts of leaves showed 83.3% callus formation rate at $NAA\;2\;mgL^{-1}$ and BA 4 mgL-1 treatment in E. elegans. The shoot induction rate from callus was highest at $NAA\;0.1\;mgL^{-1}$ and $BA\;3\;mgL^{-1}$ treatment in E. laui and $NAA\;0.3\;mgL^{-1}$ in E. elegans. In addition, the number of shoots formation was 10.4 shoots high in $NAA\;1\;mgL^{-1}$ and $BA\;1\;mgL^{-1}$ treatment in E. laui and 12.0 shoots in most effective $NAA\;1\;mgL^{-1}$ and $BA\;0.1\;mgL^{-1}$ treatment in E. elegans. In the case of acclimatization of regenerated plant, growth characteristics did not show any significant difference (35 ~ 55%) shading with respect to the different ratio of substrate mixture, and it was determined that would be appropriate considered plant height and appearance preference of E. laui and E. elegans. It was established that the optimization of culture condition was responsible for the mass propagation in vitro cultures of E. laui and E. elegans.

Production of Poly(3-hydroxybutyrate) Using Waste Frying Oil (Waste frying oil를 사용한 Poly(3-Hydroxybutyrate) 생합성)

  • Kim, Tae-Gyeong;Lee, Woosung;Gang, Seongho;Kim, Jong-Sik;Chung, Chung-Wook
    • Journal of Life Science
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    • v.29 no.1
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    • pp.76-83
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    • 2019
  • In this study, the optimal growth and poly(3-hydroxybutyrate) (PHB) biosynthesis of Pseudomonas sp. EML2 were established using waste frying oil (WFO) as a cheap carbon source. The fatty acid composition of WFO and fresh frying oil (FFO) were analyzed by gas chromatography. The unsaturated and saturated fatty acid contents of the FFO were 82.6% and 14.9%, respectively. These contents changed in the WFO. The compositional change in the unsaturated fatty acid content in the WFO was due to a change in its chemical and physical properties resulting from heating, an oxidation reaction, and hydrolysis. The maximum dry cell weight (DCW) and PHB yield (g/l) of the isolated strain Pseudomonas sp. EML2 were confirmed under the following culture conditions: 30 g/l of WFO, 0.5 gl of $NH_4Cl$, pH 7, and $20^{\circ}C$. Based on this, the growth and PHB yield of Pseudomonas sp. EML2 were confirmed by 3 l jar fermentation. After the cells were cultured in 30 g/l of WFO for 96 h, the DCW, PHB content, and PHB yield of Pseudomonas sp. EML2 were 3.6 g/l, 73 wt%, and 2.6 g/l, respectively. Similar results were obtained using 30 g/l of FFO as a carbon source control. Using the FFO, the DCW, PHB content, and PHB yield were 3.4 g/l, 70 wt%, and 2.4 g/l, respectively. Pseudomonas sp. EML2 and WFO may be a new candidate and substrate, respectively, for industrial production of PHB.

A Study on Status Analysis for Advancement iNto Agricultural Sector in Central Asia (중앙아시아 농업분야 진출을 위한 현황분석 - 우즈베키스탄, 카자흐스탄, 키르기즈스탄 중심으로 -)

  • Park, Dong-Jin;Jo, Sung-Ju;Park, Jeong-Woon;Sa, Soo-Jin;Hong, Jung-Sik;Lee, Dong-Jin
    • Journal of the Korean Society of International Agriculture
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    • v.30 no.4
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    • pp.328-338
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    • 2018
  • Central Asia (Uzbekistan, Kazakhstan, Kyrgyzstan) is a hot and arid continental climate, with most areas (68%) consisting of barren vegetation, desert, and meadows. The main agricultural areas for crop production include irrigated farmland, non-irrigated farmland, grassland, prairie and mountain. We are experiencing climate change with recent climate variability increasing. Agriculture is one of major economic sectors and provides a means of livings for the rural population of Central Asia, especially the poor. In the past two decades, Central Asia has experienced a high population growth rate, with Kazakhstan at 16.8%, Uzbekistan at 34.5% and Kyrgyzstan at 28.4%. As a major industry, Kazakhstan has the largest share of exports of agricultural products followed by petroleum, mineral resources, steel, and chemicals. Uzbekistan is the fifth largest cotton exporter as well as the sixth largest cotton producer in the world. Kyrgyzstan exports ores, stones, cultured pearls, and minerals. These three countries are rich in mineral resources, agricultural products, and energy resources. However, not only do they have difficulties in economic development due to the weakness of logistics and industrial infrastructure, but they also have imperceptible cooperation and investment among countries due to insufficient research and development. Through this study, we will investigate national outlook, economic indicators, major agricultural products, import and export status, and agricultural technology cooperation status, and study how Korean agricultural industry advances into these countries through SWOT analysis. Through this, we hope to contribute to the basic data of Central Asian studies and cooperation and investment in agriculture in each country. In addition, in order to increase cooperative exchange and investment in these countries, we will prepare a Central Asia logistics hub for the rapidly changing interKorean railroad era.

Genetic Identification and Biochemical Characteristics of Edwardsiella Strains Isolated from Freshwater Fishes Cultured in Korea (내수면 양식 어류에서 분리된 Edwardsiella 속 균주들의 유전학적 동정 및 생화학적 특성)

  • Jang, Mun Hee;Kim, Keun-Yong;Lee, Yu Hee;Oh, Yun Kyung;Lee, Jeong-Ho;Song, Jun-Young
    • Journal of fish pathology
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    • v.33 no.2
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    • pp.111-118
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    • 2020
  • The genus Edwardsiella belonging to the family Enterobacteriaceae is a member of Gram-negative rod-shaped bacteria that cause disease in diverse aquatic organisms such as fish, amphibians and reptiles as well as avians and mammals including human throughout the world. This genus had been composed of three species, E. hoshinae, E. ictaluri and E. tarda, but recent researches erected two novel species, E. anguillarum and E. piscicida that were conventionally identified as E. tarda. In this study, we isolated seven strains belonging to the genus Edwardsiella from freshwater fishes that had been reared at inland fish farms in South Korea and investigated their biochemical characteristics and molecular phylogenetic relationships. The seven strains showed typical characteristics of four Edwardsiella species, E. anguillarum, E. ictaluri, E. piscicida and E. tarda, by biochemical analyses of Gram staining, indole and hydrogen sulfide (H2S) production, and API (Analytic Profile Index) 20E test. Molecular phylogenetic analyses inferred from DNA sequence data of both 16S ribosomal RNA (rRNA) and DNA gyrase subunit B (gyrB) genes were congruent with the biochemical characteristics. As a result, both biochemical and molecular phylogenetic analyses identified four strains isolated from three Anguilla species as E. anguillarum, E. piscicida and E. tarda, two strains from Pelteobagrus fulvidraco and Silurus asotus as E. ictaluri, and one strain from Moroco oxycephalus as E. piscicida. In this study, we isolated and successfully identified recently newly erected species, E. anguillarum and E. piscicida in addition to historically notorious pathogenic species, E. ictaluri and E. tarda. In the future study, systematic and comprehensive monitoring of the four Edwardsiella species are required for studying differences in pathogenicity among freshwater fishes.

Anti-listeria Activity of Lactococcus lactis Strains Isolated from Kimchi and Characteristics of Partially Purified Bacteriocins (김치에서 분리한 Lactococcus lactis 균주의 항리스테리아 활성 및 부분 정제된 박테리오신의 특성)

  • Son, Na-Yeon;Kim, Tae-Woon;Yuk, Hyun-Gyun
    • Journal of Food Hygiene and Safety
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    • v.37 no.2
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    • pp.97-106
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    • 2022
  • Listeria monocytogenes (L. monocytogenes) is one of gram-positive foodborne pathogens with a very high fatality rate. Unlike most foodborne pathogens, L. monocytogenes is capable of growing at low temperatures, such as in refrigerated foods. Thus, various physical and chemical prevention methods are used in the manufacturing, processing and distribution of food. However, there are limitations to the methods such as possible changes to the food quality and the consumer awareness of synthetic preservatives. Thus, the aim of this study was to evaluate the anti-listeria activity of lactic acid bacteria (LAB) isolated from kimchi and characterize the bacteriocin produced by Lactococcuslactis which is one of isolated strains from kimchi. The analysis on the anti-listeria activity of a total of 36 species (Lactobacillus, Weissella, Lactobacillus, and Lactococcus) isolated from kimchi by the agar overlay method revealed that L. lactis NJ 1-10 and NJ 1-16 had the highest anti-listeria activity. For quantitatively analysis on the anti-listeria activity, NJ 1-10 and NJ 1-16 were co-cultured with L. monocytogenes in Brain Heat Infusion (BHI) broth, respectively. As a result, L. monocytogenes was reduced by 3.0 log CFU/mL in 20 h, lowering the number of bacteria to below the detection limit. Both LAB strains showed anti-listeria activity against 24 serotypes of L. monocytogenes, although the sizes of clear zone was slightly different. No clear zone was observed when the supernatants of both LAB cultures were treated with proteinase-K, indicating that their anti-listerial activities might be due to the production of bacteriocins. Heat stability of the partially purified bacteriocins of NJ 1-10 and NJ 1-16 was relatively stable at 60℃ and 80℃. Yet, their anti-listeria activities were completely lost by 60 min of treatment at 100℃ and 15 min of treatment at 121℃. The analysis on the pH stability showed that their anti-listeria activities were the most stable at pH 4.01, and decreased with the increasing pH value, yet, was not completely lost. Partially purified bacteriocins showed relatively stable anti-listeria activities in acetone, ethanol, and methanol, but their activities were reduced after chloroform treatment, yet was not completely lost. Conclusively, this study revealed that the bacteriocins produced by NJ 1-10 and NJ 1-16 effectively reduced L. monocytogenes, and that they were relatively stable against heat, pH, and organic solvents, therefore implying their potential as a natural antibacterial substance for controlling L. monocytogenes in food.