• Title/Summary/Keyword: ITS1 sequencing

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Breeding of the Scab-Resistant Pear Cultivar 'Greensis' (배 검은별무늬병 저항성 품종 '그린시스' 육성)

  • Kim, Yoon-Kyeong;Kang, Sam-Seok;Won, Kyung-Ho;Shin, Il-Sheob;Cho, Kwang-Sik;Ma, Kyeong-Bok;Kim, Myung-Su;Choi, Jang-Jeon;Choi, Jin-Ho
    • Horticultural Science & Technology
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    • v.34 no.4
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    • pp.655-661
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    • 2016
  • To develop scab-resistant pear (Pyrus spp.) varieties with fruits that are as crisp and juicy as Asian pears, a cross was made between 'Whangkeumbae' and 'Bartlett' varieties (P. pyrifolia ${\times}$ P. communis) at the Pear Research Institute of the National Institute of Horticultural & Herbal Science, Rural Development Administration, in 1994. Among the 285 seedlings, 'Greensis' was first selected in 2006 for its good eating quality and named in 2012 after regional adaptation tests in nine regions and ten experimental plots from 2007 to 2012. The tree showed a vigorous growth habit and semi-spreading characteristics, like 'Whangkeumbae'. The optimum fruit harvest date was also around Sept. 26 and fruit was round in shape and green in skin color at maturity. Average fruit weight was 470g, and the soluble solids content was $12.4^{\circ}Brix$. The flesh was very crisp and juicy, and had good eating quality. Its' leaf size was similar with 'Bartlett' and smaller than 'Whangkeumbae'. The average of full bloom date of 'Greensis' was determined as Apr. 26, which was six days later than 'Whangkeumbae' and similar with 'Bartlett'. S genotypes of 'Greensis' were identified as $S_4S_e$ by S-allele PCR product sequencing analysis. It seems that the $S_4$ allele was inherited from 'Whangkeumbae' and the Se allele from 'Bartlett'. 'Greensis' displayed strong resistance to scab disease caused by Venturia nashicola, similar to European pear cultivars like 'Beurre Hardy' and, 'Conference'. 'Greensis' was also highly resistant to black leaf spot (Alternaria kikuchiana) in the field

Fusaric Acid Production in Fusarium oxysporum Transformants Generated by Restriction Enzyme-Mediated Integration Procedure (Restriction Enzyme-Mediated Integration 방법으로 확보한 Fusarium oxysporum 형질전환체의 후자리산 생성능 분석)

  • Lee, Theresa;Shin, Jean Young;Son, Seung Wan;Lee, Soohyung;Ryu, Jae-Gee
    • Research in Plant Disease
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    • v.19 no.4
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    • pp.254-258
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    • 2013
  • Fusaric acid (FA) is a mycotoxin produced by Fusarium species. Its toxicity is relatively low but often associated with other mycotoxins, thus enhancing total toxicity. To date, biosynthetic genes or enzymes for FA have not been identified in F. oxysporum. In order to explore the genetic element(s) for FA biosynthesis, restriction enzyme mediated integration (REMI) procedure as an insertional mutagenesis was employed using FA producing-F. oxysporum strains. Genetic transformation of two F. oxysporum strains by REMI yielded more than 7,100 transformants with efficiency of average 3.2 transformants/${\mu}g$ DNA. To develop a screening system using phytotoxicity of FA, eleven various grains and vegetable seeds were tested for germination in cultures containing FA: Kimchi cabbage seed was selected as the most sensitive host. Screening for FA non-producer of F. oxysporum was done by growing each fungal REMI transformant in Czapek-Dox broth for 3 weeks at $25^{\circ}C$ then observing if the Kimchi cabbage seeds germinated in the culture filtrate. Of more than 5,000 REMI transformants screened, fifty-three made the seeds germinated, indicating that they produced little or fewer FA. Among them, twenty-six were analyzed for FA production by HPLC and two turned out to produce less than 1% of FA produced by a wild type strain. Sequencing of genomic DNA regions (252 bp) flanking the vector insertion site revealed an uncharacterized genomic region homologous (93%) to the F. fujikuroi genome. Further study is necessary to determine if the vector insertion sites in FA-deficient mutants are associated with FA production.

A Case of Adult-onset Type II Citrullinemia Confirmed by Mutation of SLC25A13 (SLC25A13 유전자 돌연변이로 확진된 성인형 제 2형 시트룰린혈증 1례)

  • Jeung, Min Sub;Yang, Aram;Kim, Jinsup;Park, Hyung-Doo;Lee, Heon Ju;Jin, Dong-Kyu;Cho, Sung Yoon
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.16 no.1
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    • pp.34-41
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    • 2016
  • Adult-onset type II citrullinemia (CTLN2) is characterized by episodes of neurologic symptoms associated with hyperammonemia leading to disorientation, irritability, seizures, and coma. CTLN2 is distinct from classical citrullinemia, which is caused by a mutation of the argininosuccinic acid synthetase (ASS) gene. The serum citrulline level is elevated, while the activity of ASS in liver tissue is decreased. CTLN2 is known to have a poor prognosis if the proper treatment is not taken. We reported a female aged 37 years who developed recurrent attacks of altered consciousness, aberrant behavior, and vomiting. We initially suspected the patient had CTLN2 because of the signs of hyperammonemic encephalopathy, such as altered mentality, memory disturbance, and aberrant behaviors provoked by exercise-induced stress and excessive intravenous amino acid administration. Through her peculiar diet preferences and laboratory findings that included hyperammonemia and citrullinemia, we diagnosed the patient as CTLN2, and SLC25A13 sequencing revealed known compound heterozygous mutations (IVS11+1G>A, c.674C> A). Her parents were heterozygous carriers, and we identified that her older sister had the same mutations. The older sister had not experienced any episodes of hyperammonemia, but she had peculiar diet preferences. The patient and her sister have been well with conservative management. When considering the clinical course of CTLN2, it was meaningful that the older sister could be diagnosed early in an asymptomatic period and that preemptive treatment was employed. Through this case, CTLN2 should be considered in adults who present symptoms of hyperammonemic encephalopathy without a definite etiology. Because of its rare incidence and similar clinical features, CTLN2 is frequently misdiagnosed as hepatic encephalopathy, and it shows a poor prognosis due to the lack of early diagnosis and proper treatment. A high-carbohydrate diet, which is usually used to treat other urea cycle defects, can also exaggerate the clinical course of CTLN2, so proper metabolic screening tests and genetic studies should be performed.

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Identification of a SNP in Chicken CaSR Gene and Its Effect on Economic Traits (닭의 CaSR 유전자내 단일 염기 변이 탐색 및 경제 형질간의 연관성 분석)

  • Hong, Y.S.;Oh, J.D.;Lee, J.H.;Kong, H.S.;Choi, C.H.;Lee, S.S.;Jeon, G.J.;Lee, H.K.
    • Korean Journal of Poultry Science
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    • v.34 no.2
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    • pp.151-156
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    • 2007
  • The Function of the calcium sensing receptor (CaSR) is to control calcium levels by altering PTH (parathyroid hormone) secretion and renal calcium resorption. The influence of calcium on the basal and stimulated release of several hormones from chicken pituitary glands has been determined in vitro. The objective of this study was to identify SNP in chicken CaSR gene and to investigate the effect of the SNP on economic traits. The sequencing analysis method was used to identify nucleotide polymorphisms within chicken CaSR gene. This study identified SNP at position 1949 bp(Genebank accession No : XM_416491) in the exon 1. The SNP changed the amino acid to alanine(GCC) from serine(TCC). This SNP showed three genotypes, AA, AS and SS by digestion with the restriction enzyme NcoⅠ using the PCR-RFLP method. The A963S showed significant effect only on the first lay day (P<0.05) in Leghorn population. Leghorn with the genotype AA had significantly faster the first lay day(137.6) than the genotype AS(143.0, P<0.05). Also, the A963S showed significant effect only on the first lay day(P<0.05) and mean of egg weight(P<0.05) in KNC population. KNC with the genotypes AA ans AS had significantly faster the first lay day (151.0 and 152.6, respectively) than the genotype SS(159.4, P<0.05). And the genotypes SS had significantly heavier the mean of egg weight(50.4 kg, P<0.05) than the genotype AA ans AS (47.5 and 47.8 kg, respectively). According to result of this study, an a allele of the A963S was found to have a significant effect on the first lay day. It will be possible to use this SNP marker on selecting chicken to improve the first lay day.

Comparative Transcriptome Analysis of the Response of Two Lines of Rapeseed (Brassica napus L.) to Cold Stress (유채 두 계통에서 저온 스트레스에 반응하는 전사체 발현 비교 분석)

  • Lee, Ji-Eun;Kim, Kwang-Soo;Cha, Young-Lok;An, Da-Hee;Byun, Jong-Won;Kang, Yong-Ku
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.66 no.1
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    • pp.37-71
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    • 2021
  • Rapeseed is a typical winter crop, and its freezing stress tolerance is a major feature for winter survival. Therefore, it is important to comprehend clearly the physical and molecular mechanisms of rapeseed under freezing stress conditions. This study investigates the physical and transcriptome changes of two rapeseed lines, 'J8634-B-30' and 'EMS26', under cold acclimation and freezing temperature treatments. The proline content of 'J8634-B-30' at 5 ℃ increased 8.7-fold compared to that before treatment, and there was no significant change in that of 'EMS26' RNA-sequencing analysis revealed 5,083 differentially expressed genes (DEGs) of 'J8634-B-30' under cold acclimation condition. Among the genes, 2,784 (54.8%) were up-regulated and 2,299 (45.2%) were down-regulated. The DEGs of 'EMS26' under cold acclimation condition were 5,831 genes, and contained 2,199 up-regulated genes (37.7%) and 3,632 down-regulated genes (62.3%). Among them, only DEGs annotated in the cold response-related signaling pathways were selected, and their expression in the two rapeseed lines was compared. Comparative DEGs analysis indicated that cold response related signaling pathways are proline metabolism and ABA (Abscisic acid) signaling. And ICE (Inducer of CBF expression) - CBF (C-repeat-binding factor) - COR (Cold-regulated) signaling were the significantly differentially expressed transcripts in the two rapeseed lines. The major induced transcripts of 'J8634-B-30' induced P5CS (Δ'-pyrroline-5-carboxylate synthetase), which is related to proline biosynthesis, PYL (pyrabactin resistance-like protein, ABA receptor) and COR413 (cold-regulated 413 plasma membrane 1). In conclusion, these result provide a foundation for understanding the mechanisms of freezing stress tolerance in rapeseeds. Further functional studies should be performed on the freezing stress-related genes identified in this study, which can contribute to the transgenic and molecular breeding for freezing stress tolerance in rapeseed.

Isolation and Identification of Competitive Fungi on Medium for Black Wood Ear Mushroom in Korea and In Vitro Selection of Potential Biocontrol Agents (목이버섯 배지 오염 곰팡이균의 분리, 동정 및 생물학적 방제제 선발)

  • Seoyeon Kim;Miju Jo;Sunmin An;Jiyoon Park;Jiwon Park;Sungkook Hong;Jiwoo Kim;Juhoon Cha;Yujin Roh;Da Som Kim;Mi jin Jeon;Won-Jae Chi;Sook-Young Park
    • Research in Plant Disease
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    • v.30 no.1
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    • pp.66-77
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    • 2024
  • Black wood ear mushroom (Auricularia auricula-judae) is one of the most economically important mushrooms in China, Japan, and Korea. The cultivation of wood ear mushrooms on artificial substrates is more efficient in terms of time and cost compared with their natural growth on trees. However, if the substrate cultivation is infected by fast-growing fungi, the relatively slow-growing ear mushroom will be outcompeted, leading to economic losses. In this study, we investigated the competitive fungal isolates from substrates infected with fast-growing fungi for the cultivation of ear mushrooms in Jangheung and Sunchon, Korea. We collected 54 isolates and identified them by sequencing their internal transcribed spacer region with morphological identification. Among the isolates, the dominant isolates were Trichoderma spp. (92.6%), Penicillium spp. (5.6%), and Talaromyces sp. (1.8%). To find an appropriate eco-friendly biocontrol agent, we used five Streptomyces spp. and Benomyl, as controls against Trichoderma spp. and Penicillium spp. Among the six Streptomyces spp., Streptomyces sp. JC203-3 effectively controlled the fungi Trichoderma spp. and Penicillium spp., which pose a significant problem for the substrates of black wood ear mushrooms. This result indicated that this Streptomyces sp. JC203-3 can be used as biocontrol agents to protect against Trichoderma and Penicillium spp.

Identification and Characterization of Lactobacillus salivarius subsp. salivarius from Korean Feces

  • Bae, Hyoung-Churl
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2004.05a
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    • pp.89-119
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    • 2004
  • This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

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Development and Characterization of EMS-induced Mutants with Enhanced Salt Tolerance in Silage Maize (EMS 유도 내염성 증진 사료용 옥수수 돌연변이체 선발 및 특성 분석)

  • Cho, Chuloh;Kim, Kyung Hwa;Seo, Mi-Suk;Choi, Man-Soo;Chun, Jaebuhm;Jin, Mina;Kim, Dool-Yi
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.4
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    • pp.406-415
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    • 2020
  • Maize (Zea mays L.) is one of the most valuable agricultural crops and is grown under a wide spectrum of environmental conditions. However, maize is moderately sensitive to salt stress, and soil salinity is a serious threat to its production worldwide. In this study, we used ethyl methane sulfonate (EMS) to generate salt-tolerant silage maize mutants. We screened salt-tolerant lines from 203 M3 mutant populations by evaluating the morphological phenotype after salt stress treatment and selected the 140ES91 line. The 140ES91 mutant showed improved plant growth as well as higher proline content and leaf photosynthetic capacity compared with those of wild-type plants under salt stress conditions. Using whole-genome re-sequencing analysis, 1,103 single nucleotide polymorphisms and 71 insertions or deletions were identified as common variants between KS140 and 140ES91 in comparison with the reference genome B73. Furthermore, the expression patterns of three genes, which are involved in salt stress responses, were increased in the 140ES91 mutant under salt stress. Taken together, the mutant line identified in our study could be used as an improved breeding material for transferring salt tolerance traits in maize varieties.

The Proportion of Rifabutin-susceptible Strains among Rifampicin-resistant Isolates and Its Specific rpoB Mutations (한국에서 분리된 리팜핀 내성 균주에서의 리파부틴 감수성 정도 및 관련 rpoB 유전자 돌연변이의 특성에 관한 연구)

  • Lew, Woo Jin;Park, Young Kil;Kim, Hee Jin;Chang, Chulhun;Bai, Gill Han;Kim, Sung Kyu
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.3
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    • pp.257-265
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    • 2005
  • Background : Rifabutin (ansamycin) is a spiro-piperidyl rifamycin, which is highly active against Mycobacterium tuberculosis. It has been found that some clinical isolates of tubercle bacilli that are resistant to rifampicin are susceptible to rifabutin, with some patients with multi-drug resistant pulmonary tuberculosis having shown favorable clinical and bacteriological responses to the rifabutin. This study was conducted to find the proportion of rifabutin-susceptible strains among rifampicin-resistant isolates from Korean MDR-TB patients, and investigate the presence of specific rpoB mutations, which may confer resistance to rifampicin, but not to rifabutin. Methods : 201 rifampicin-resistant and 50 pan-susceptible M. tuberculosis isolates were randomly selected for this study. The isolates were retested at rifampicin and rifabutin concentrations of 0, 20, 40 and $80{\mu}g/ml$, respectively. The isolates that grew at and/or over a rifabutin concentration of $20{\mu}g/ml$ were judged rifabutin-resistant. The rpoB gene was extracted from the isolates, and then amplified for direct sequencing to investigate specific rpoB mutations that conferred rifabutin- susceptibility but rifampicin-resistance. Results : Out of the 201 rifampicin-resistant M. tuberculosis, 41 strains (20.4%) were susceptible to rifabutin using the absolute concentration method on Lowenstein-Jensen media. The rpoB mutation types that showed susceptibility to rifabutin were Leu511Pro, Ser512Arg, Gln513Glu, Asp516Ala, Asp516Gly, Asp516Val, Asp516Tyr, Ser522Leu, His526Asn, His526Leu, His526Cys, Arg529Pro and Leu533Pro. A reverse hybridization technique was able to detect 92.5% of the rifabutin-susceptible isolates, with a specificity of 96.1% among 195 M. tuberculosis isolates with the rpoB mutation. Conclusions : Around 20% of the rifampicin-resistant isolates in Korea showed susceptibility to rifabutin, which was associated with some specific mutations of rpoB. Rifabutin could be used for the treatment of MDR-TB patients, especially when drug susceptibility testing reveals susceptibility to rifabutin.

Anti-inflammatory Activity of Antimicrobial Peptide Papiliocin 3 Derived from the Swallowtail Butterfly, Papilio xuthus (호랑나비 유래 항균 펩타이드 파필리오신 3의 항염증 활성)

  • Shin, Yong Pyo;Lee, Joon Ha;Kim, In-Woo;Seo, Minchul;Kim, Mi-Ae;Lee, Hwa Jeong;Baek, Minhee;Kim, Seong Hyun;Hwang, Jae Sam
    • Journal of Life Science
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    • v.30 no.10
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    • pp.886-895
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    • 2020
  • The development of novel peptide antibiotics with potent antimicrobial activity and anti-inflammatory activity is urgently needed. In a previous work, we performed an in-silico analysis of the Papilio xuthus transcriptome to identify putative antimicrobial peptides and identified several candidates. In this study, we investigated the antibacterial and anti-inflammatory activities of papiliocin 3, which was selected bioinformatically based on its physicochemical properties against bacteria and mouse macrophage Raw264.7 cells. Papiliocin 3 showed antibacterial activities against E. coli and S. aureus without inducing hemolysis and decreased the nitric oxide production of the lipopolysaccharide-induced Raw264.7 cells. Moreover, ELISA and Western blot analysis revealed that papiliocin 3 reduced the expression levels of pro-inflammatory enzymes, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and prostaglandin E2 (PGE2). In addition, we examined whether papiliocin 3 could inhibit the expression of pro-inflammatory cytokines (interleukin-6 and interleukin-1β) in LPS-induced Raw264.7 cells. We found that papiliocin 3 markedly reduced the expression level of cytokines through the regulation of mitogen-activated protein kinases (MAPK) and nuclear factor kappa B (NF-κB) signaling. We also confirmed that papiliocin 3 binds to bacterial cell membranes via a specific interaction with lipopolysaccharides. Collectively, these findings suggest that papiliocin 3 could be a promising molecule for development as a novel peptide antibiotic.