• 제목/요약/키워드: Bacillibactin

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Bacillus subtilis AH18의 고추역병 방제능과 $Siderophore_{AH18}$의 구조분석 (Structural Identification of $Siderophore_{AH18}$ from Bacillus subtilis AH18, a Biocontrol agent of Phytophthora Blight Disease in Red-pepper)

  • 우상민;김상달
    • 한국미생물·생명공학회지
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    • 제36권4호
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    • pp.326-335
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    • 2008
  • Bacillus subtilis AH18균주는 auxin, siderophore 그리고 cellulase를 동시에 생산하는 PGPR 균주이자 생물방제균주로 항진균성 siderophore의 특성을 확인한 결과 catechol type의 siderophore로 확인하였다. $Siderophore_{AH18}$ 의 정제는 Amberlite XAD-2, sephadex LH-20 column chromatography 그리고 HPLC를 통해 정제 및 정제여부를 확인하였으며, GC-MS, $^1H$-NMR, 그리고 $^{13}C$-NMR을 통해 구조 및 분자량을 확인하였다. 그 결과 B. subtilis AH18이 생산하는 siderophore은 분자량 883의 bacillibactin임을 확인하였으며, 포자발아억제활성을 나타냄을 확인하였다. B. subtilis AH18은 P. capsici에 의한 고추역병을 효과적으로 방제하였으며(방제력 55%), 이는 bacillibactin에 의한 효과가 포함되리라 추측된다.

액체크로마토그래피-사중극 비행시간형 탠덤질량분석기를 이용한 Bacillus mojavensis 균주 속 사이드로포어 규명 (Determination of Siderophore from Bacillus Mojavensis Using Liquid Chromatography quadrupole Time-of-flight Tandem Mass Spectrometry)

  • 천해인;여미선;김강민;강재선;표재성
    • 생명과학회지
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    • 제29권2호
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    • pp.198-201
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    • 2019
  • Bacillus mojavensis를 이용한 항진균성 및 식물 생장 촉진 활성이 최근 보고되었다. 이런 활성은 사이드로포어의 일반적 특성과 일치하여, 본 연구에서는 Bacillus mojavensis가 생산하는 사이드로포어를 고체상추출 카트리지와 액체크로마토그래피-사중극 비행시간형 탠덤 질량분석기를 이용하여 규명하였다. Bacillus mojavensis를 페놀 배지에서 16시간 동안 배양하고 동결 건조 시킨 후, 물에 용해시켜 고상추출 카트리지에 로딩하였다. 카트리지는 5% 메탄올로 세척하고 2% 포름산을 이용해 용출 시켰다. 용출액은 메탄올에 재용해 후 분석을 하였다. Bacillus mojavensis의 사이드로포어는 질량 스펙트럼의 결과를 바탕으로 882.2556의 분자량을 갖는 카테콜타입의 사이드로포어 중 하나인 bacillibactin으로 확인되었다. 이 사이드로포어 분석은 Bacillus 연구 및 기능성 식품 그리고 Bacillus mojavensis의 약학 응용 분야에 큰 기여를 할 것으로 예상된다.

식물생육촉진, 항균 및 저항성 유도 효과를 나타내는 내생세균 Bacillus velezensis YC7010의 유전체 염기서열 (Complete genome sequence of Bacillus velezensis YC7010, an endophytic bacterium with plant growth promoting, antimicrobial and systemic resistance inducing activities in rice)

  • 라시드엠디하룬;황정현;정영륜
    • 미생물학회지
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    • 제53권4호
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    • pp.329-331
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    • 2017
  • 식물 내생 세균인 Bacillus velezensis YC7010은 벼의 병원성 세균과 곰팡이에 대해 전신유도저항성을 일으키며, 식물생육촉진 및 항생작용의 특징을 가지고 있다. 대한민국 진주지역 벼의 근권에서 분리된 B. velezensis YC7010의 유전체는 3,790개의 단백질-암호화 유전자(86 tRNAs와 27 rRNA 유전자)로 구성되어 있으며, 3,975,683 염기쌍의 환상 염색체이다. 유전체 분석을 통해 식물의 발달과 생육을 촉진하는 휘발성 화합물, 식물호르몬 생산, surfactin, plipapastatin, bacillibactin, bacillaene 같은 활성 화합물의 생합성, 식물 내부정착 및 군집화와 관련된 유전자들이 유전체에 존재하는 것을 확인하였다

Evaluation and Genome Mining of Bacillus stercoris Isolate B.PNR1 as Potential Agent for Fusarium Wilt Control and Growth Promotion of Tomato

  • Rattana Pengproh;Thanwanit Thanyasiriwat;Kusavadee Sangdee;Juthaporn Saengprajak;Praphat Kawicha;Aphidech Sangdee
    • The Plant Pathology Journal
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    • 제39권5호
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    • pp.430-448
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    • 2023
  • Recently, strategies for controlling Fusarium oxysporum f. sp. lycopersici (Fol), the causal agent of Fusarium wilt of tomato, focus on using effective biocontrol agents. In this study, an analysis of the biocontrol and plant growth promoting (PGP) attributes of 11 isolates of loamy soil Bacillus spp. has been conducted. Among them, the isolates B.PNR1 and B.PNR2 inhibited the mycelial growth of Fol by inducing abnormal fungal cell wall structures and cell wall collapse. Moreover, broad-spectrum activity against four other plant pathogenic fungi, F. oxysporum f. sp. cubense race 1 (Foc), Sclerotium rolfsii, Colletotrichum musae, and C. gloeosporioides were noted for these isolates. These two Bacillus isolates produced indole acetic acid, phosphate solubilization enzymes, and amylolytic and cellulolytic enzymes. In the pot experiment, the culture filtrate from B.PNR1 showed greater inhibition of the fungal pathogens and significantly promoted the growth of tomato plants more than those of the other treatments. Isolate B.PNR1, the best biocontrol and PGP, was identified as Bacillus stercoris by its 16S rRNA gene sequence and whole genome sequencing analysis (WGS). The WGS, through genome mining, confirmed that the B.PNR1 genome contained genes/gene cluster of a nonribosomal peptide synthetase/polyketide synthase, such as fengycin, surfactin, bacillaene, subtilosin A, bacilysin, and bacillibactin, which are involved in antagonistic and PGP activities. Therefore, our finding demonstrates the effectiveness of B. stercoris strain B.PNR1 as an antagonist and for plant growth promotion, highlighting the use of this microorganism as a biocontrol agent against the Fusarium wilt pathogen and PGP abilities in tomatoes.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 한국균학회 2015년도 춘계학술대회 및 임시총회
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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