• Title/Summary/Keyword: Bacterial whole cells

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Genomic Analysis of Halotolerant Bacterial Strains Martelella soudanensis NC18T and NC20

  • Jung-Yun Lee;Dong-Hun Kim
    • Journal of Microbiology and Biotechnology
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    • v.32 no.11
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    • pp.1427-1434
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    • 2022
  • Two novel, halotolerant strains of Martelella soudanensis, NC18T and NC20, were isolated from deep subsurface sediment, deeply sequenced, and comparatively analyzed with related strains. Based on a phylogenetic analysis using 16S rRNA gene sequences, the two strains grouped with members of the genus Martelella. Here, we sequenced the complete genomes of NC18T and NC20 to understand the mechanisms of their halotolerance. The genome sizes and G+C content of the strains were 6.1 Mb and 61.8 mol%, respectively. Moreover, NC18T and NC20 were predicted to contain 5,849 and 5,830 genes, and 5,502 and 5,585 protein-coding genes, respectively. Both strains contain the identically predicted 6 rRNAs and 48 tRNAs. The harboring of halotolerant-associated genes revealed that strains NC18T and NC20 might tolerate high salinity through the accumulation of potassium ions in a "salt-in" strategy induced by K+ uptake protein (kup) and the K+ transport system (trkAH and kdpFABC). These two strains also use the ectoine transport system (dctPQM), the glycine betaine transport system (proVWX), and glycine betaine uptake protein (opu) to accumulate "compatible solutes," such as ectoine and glycine betaine, to protect cells from salt stress. This study reveals the halotolerance mechanism of strains NC18T and NC20 in high salt environments and suggests potential applications for these halotolerant and halophilic strains in environmental biotechnology.

Extraction and verification of highly immunologenic antigen for diagnosis of bovine brucellosis (소 브루셀라병 진단용 고면역원성 항원의 추출과 검증)

  • Bae, Jae-Hyung;Jo, Sang-Rae;Jeong, Eun-Hui;Jang, Eun-Hui;Kim, Seong-Eun;Kwon, Hui-Nyeong;Park, Dong-Yeop;Lee, Kuk-Cheon
    • Korean Journal of Veterinary Service
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    • v.33 no.2
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    • pp.135-141
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    • 2010
  • Bovine brucellosis, an important zoonosis, is diagnosed with serological tests such as the RBT, TAT using inactivated whole bacterial cells or bacterial lipopolysaccharide (LPS) antigen in Korea. However, a strong cross-reaction between Brucella spp. and Yersinia enterocolitica O9 in these tests has seriously complicated the diagnosis of animal brucellosis because Brucella spp. shares common antigenic determinants with Y. enterocolitica O9 in the smooth LPS region. In this study, Brucella-field strains were isolated from Brucellapositive Hanwoo in Kimhae, Korea and outer membrane protein (omp) which has low cross-reaction with Y. enterocolitica O9 and high immunogenicity was extracted from the field strains Then we compared ELISA using the extract with RBT-TAT. Fifteen field strains were isolated from 47 supra-mammary-lymph nodes, which were collected from 18 farms. Isolation rate was 32%. Brucella-specific antigen was identified by performing SDS-PAGE or Western blotting on extracted omp with at 0.5% n-lauroylsarcosine One hundred and ninety-two serum-samples were used in the experiment: 142 negative and 50 positive samples verified by RBT-TAT. According to ELISA results, 127 samples were negative and 15 appeared positive among 142 negatives by RBT-TAT, while 42 samples were positive and 8 were negative among 50 positives by RBT-TAT. Therefore, it showed 89.4% of specificity and 84% of sensi-tivity. Through the current experiments, we could set up an ELISA based on the omp which has low cross-reaction and high immunogenicity and concluded that the omp could be a good material for accurate diagnosis of bovine brucellosis.

The Various Effects of Xylitol as a Dietary Sugar Substitute on Improving Oral Health (식이 당 대체제인 자일리톨의 구강건강 증진에 미치는 다양한 효과)

  • Hwang, Yoon-Sook;Lee, Hu-Jang
    • Journal of Food Hygiene and Safety
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    • v.37 no.2
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    • pp.107-113
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    • 2022
  • A number of studies have been conducted to confirm the preventive effect of xylitol on dental caries as a whole or partial alternative to dietary sugars. This study reviewed the oral health effects of xylitol on the prevention mechanism of dental caries, the prevention of dental caries, the inhibition of mother-to-child transmission, and the oral health effects in the elderly based on existing studies on the oral health of xylitol. Carbohydrates and dietary sugars in food are fermented by acid-producing microorganisms in the mouth and produce dental plaque and acid, which cause dental caries. However, most dental decay-causing bacteria cannot produce acids by metabolizing xylitol. Xylitol, stored in cells as a non-metabolizable metabolite by Streptococcus mutans (S. mutans), affects bacterial glucose metabolism and inhibits bacterial growth. Xylitol consumption also reduces the amount of plaque and the population of S. mutans in both plaque and saliva. In addition, xylitol acts in the remineralization process. Xylitol has been confirmed to effectively prevent dental caries, inhibit mother-to-child transmission of MS, prevent dental caries, and increase salivary flow in the elderly. In conclusion, xylitol is an adequate sugar substitute for dental health, from infants to the elderly. For future studies, the researchers recommend reviewing the effects of xylitol on the oral and intestinal microbial environment and the side effects of excessive intake.

Immunomodulatory effect of the water extract of Aster tataricus through mitogen-activated protein kinase signaling pathway (Aster tataricus 물 추출물의 mitogen-activated protein kinase 신호 전달 경로를 통한 면역 조절 효과)

  • Lee, Chea Yeon;Park, Hyo Sung;Kong, Deok-Hoon;Kim, Young Kwan;Cho, Whajung
    • Journal of Nutrition and Health
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    • v.53 no.5
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    • pp.452-463
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    • 2020
  • Purpose: Aster tataricus (AT) is one of the Asteraceae perennial herbs used in traditional Chinese medicine. The herb contains various bioactive substances, such as flavonoids, isoflavonoids, and phenolic compounds in the roots, and exhibits a range of effects including anti-bacterial, anti-oxidant, and anti-inflammatory activities. This study compared the immunomodulatory effects of ethanol and water extracts of whole AT, except the roots, and analyzed the molecular mechanisms for the regulatory effects on cytokine secretion from THP-1 cells. Methods: The effects of AT extract on the cell viability and proliferation of THP-1 cells were analyzed using the Cell Counting Kit-8 method. The concentrations of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the cell culture supernatant of the AT-treated THP-1 cells were measured using an enzyme-linked immunosorbent assay. The protein levels of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), inhibitor of nuclear factor kappa B (IκBα), and mitogen-activated protein kinase (MAPK) phosphorylation in the cell lysates were determined by western blotting. Results: The water extract and the ethanol extract of AT did not affect the cell viability, and increased the proliferation of THP-1 cells significantly compared to the vehicle. The water extract increased the secretion of IL-1β from THP-1 cells in a dose-dependent manner, but the ethanol extract had no effect. The expression of COX-2 and iNOS protein and the phosphorylation of MAPK and Akt were induced in AT-treated cells. In addition, IκBα was degraded by AT in a concentration-dependent manner. IL-1β secretion by AT was reduced by extracellular-signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) inhibitors, while TNF-α secretion was decreased by inhibitors of ERK, p38 MAPK, and JNK. Interestingly, the p38 MAPK inhibitor increased the production of IL-1β by AT further. Conclusion: The water extract of the above-ground parts of AT contains immunomodulatory bioactive substances that stimulate immune cells through the MAPK signaling pathway.

Studies on the properties of Bordetella bronchiseptica isolated from the pig herds in Western Chungnam (충남 서부지역 돈군에서 분리된 Bordetella bronchiseptica의 성상에 대한 연구)

  • 박세종;안식욱;신인환;정태수;전무형
    • Korean Journal of Veterinary Service
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    • v.18 no.2
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    • pp.133-151
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    • 1995
  • During 2 years from Octorber 1992 to April 1994, prevalence of general respiratory diseases and atrophic rhinitis in the pig herds located in the Western Chungnam was investigated, and isolation of B. bronchiseptica was attempted for the pigs manifested with the clinical signs of atrophic rhinitis(AR). The isolates were characterized and identified in aspects of biochemical properties, antigenicity, drug sensitivity and pathogenicity. The results obtained through the experiments are summarized as follows; 1. During 2 years of investigation, the overall prevalence of the general respiratory diseases in the pi8 herds in Western Chungnam was 35.3%, consisting of 35.1% in the pig farms and 38.8% in a slaughter house. The prevalence by age groups accounts for 9.2% in adults, 44.7% in rearings and 25.3% in sucklings. By farm size, The highest prevalence of 56.5% was observed in the smallest farm with 1 to 200 heads. 2. The prevalence of clinical cases of artrophic rhinitis was recorded by 12.7% in the group that is the sows and piglets vaccinated, 28.9% in the group that is the sows only vaccinated and 39.8% in the group of the non-vaccinated groups. In the slaughter house, 53(24.8%) of 214 pigs examined exhibit the AR lesions. 3. A total of 189 strains of B. bronchiseptica were isolated from the pig herds. Isolation rates were 12.6% in the group that is the sows and piglets vaccinated, 34.1% in the group that is the sows only vaccinated and 45.7% in the group of the non-vaccinated groups. Isolation rate in the specimen from the slaughter house was 93( 43.5% ) of 214 pigs examined. Of the AR-non-vaccinated group, the piglets aged bet- ween 61 to 90 days revealed the highest isolation rate of 58.5%. 4. The titers of antibody against B. bronchiseptica were measured by tube agglutination test. The group that is the sow and piglet-vaccinated showed the highest titer of 640-2, 560 in sow and 640longrightarrow5, 120 in piglet. The group that is the sows only-vaccinated revealed 640-2, 560 in sows and 640-1, 280 in piglets. Both of the vaccinated groups showed 100% positive reaction. The group of the non-vaccinated sho-wed relatively lower titer of 0-1, 280 in both of sows and piglets. The positive rate of the sera obtained from the slaughter house was 53.3% with the antibody titer of 0-1, 280. 5. Biochemical and serological properities of 189 isolates were very similar to those of the reference B. bronchiseptical phase I type, indicating that most of isolates are B. bronchiseptica phase I type. 6. In antimicrobial drug susceptibility, 87.3% of 189 isolates was susceptible to chloramphenicol, 79.9%, to amikacin, 64.6%, to cephalothin and less than 35.4% to others. 7. In agar-gel immunodiffusion and SDS-PAGE analysis, the isolates presented the identical antigenicity and protein profiles to the reference standard strains. 8. The whole cells and bacterial filtrates of the isolates were inoculated to guinea pigs and mice. The isolates showed the hish pathogenicity and dermonecrotoxiciy.

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Relationship between Preterm Low Birth Weight and Periodontal Disease Activity in Pregnancy (임산부의 치주 질환 활성도와 조산과의 상관관계에 관한 연구)

  • Choi, Eun-Cheong;Ku, Young;Rhyu, In-Chul;Hahm, Byung-Do;Yoon, Bo-Hyun;Han, Soo-Boo;Chung, Chong-Pyoung;Choi, Sang-Mook
    • Journal of Periodontal and Implant Science
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    • v.30 no.1
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    • pp.111-120
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    • 2000
  • Purpose We designed this study for the purpose of determining the relationship between periodontal disease activity and PLBW, using the evaluation of probing pocket depth, loss of attachment, gingival index, gingival crevicular fluid amount and subgingival microflora. Methods A total of 100 volunteer mothers(mean age 30.44) at the Department of Obstetrics and Gynecology Seoul National University Hospital were selected for this study.Pregnancy outcomes were categorized into cases and controls in two ways. our definition was based on the following; Group 1 : Any PLBW cases Vs. All NBW controls Group 2 : PLBW cases Vs. NBW controls A periodontal exam was performed on the Ramfjord( #16, 21, 24, 36, 41, 44) teeth and Clinical evaluation consisted of probing pocket depth, loss of attachment, gingival index and gingival crevicular fluid amount. Subgingival plaque samples were collected by three sterile #35 paper points. The total number of anaerobic colonies and aerobic bacteria were enumerated after incubation. Antisera to P. gingivalis, P. intermedia, A. actinomycetemcomitans were produced in white rabbits with live whole cells suspensions. The specific fluorescent bacteria obtained by immunofluorescence and total cell counts obtained by dark-field microscopy were counted on four fields. The percent of each specific microorganism in the total cell count was determined. Results Any PLBW and PLBW cases showed significantly greater probing depth and attachment loss than all NBW and NBW controls. Cases group had significantly increased anaerobic bacterial counts compared with control group and no differences in the other microbes. This study confirmed that periodontal disease is a statistically significant risk factor for PLBW by investigating clinical parameters and subgingival plaque analysis.

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Sphingomonas abietis sp. nov., an Endophytic Bacterium Isolated from Korean Fir

  • Lingmin Jiang;Hanna Choe;Yuxin Peng;Doeun Jeon;Donghyun Cho;Yue Jiang;Ju Huck Lee;Cha Young Kim;Jiyoung Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.10
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    • pp.1292-1298
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    • 2023
  • PAMB 00755T, a bacterial strain, was isolated from Korean fir leaves. The strain exhibits yellow colonies and consists of Gram-negative, non-motile, short rods or ovoid-shaped cells. It displays optimal growth conditions at 20℃, 0% NaCl, and pH 6.0. Results of 16S rRNA gene-based phylogenetic analyses showed that strain PAMB 00755T was most closely related to Sphingomonas chungangi MAH-6T (97.7%) and Sphingomonas polyaromaticivorans B2-7T (97.4%), and ≤96.5% sequence similarity to other members of the genus Sphingomonas. The values of average nucleotide identity (79.9-81.3%), average amino acid identity (73.3-75.9%), and digital DNA-DNA hybridization (73.3-75.9%) were significantly lower than the threshold values for species boundaries; these overall genome-related indexes (OGRI) analyses indicated that the strain represents a novel species. Genomic analysis revealed that the strain has a 4.4-Mbp genome encoding 4,083 functional genes, while the DNA G+C content of the whole genome is 66.1%. The genome of strain PAMB 00755T showed a putative carotenoid biosynthetic cluster responsible for its antioxidant activity. The respiratory quinone was identified as ubiquinone 10 (Q-10), while the major fatty acids in the profile were identified as C18:1ω7c and/or C18:1ω6c (summed feature 8). The major polar lipids of strain PAMB 00755T were diphosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid, and phosphatidylcholine. Based on a comprehensive analysis of genomic, phenotypic, and chemotaxonomic characteristics, we proposed the name Sphingomonas abietis sp. nov. for this novel species, with PAMB 00755T as the type strain (= KCTC 92781T = GDMCC 1.3779T).

Changes in Yeast and Bacterial Flora during Fermentation and Storage of Gugija-Liriope tuber Makgeolli using PCR-DGGE (PCR-DGGE를 이용한 구기자-맥문동 막걸리의 발효 과정과 저장 기간 중 효모와 세균 균총의 변화)

  • Min, Jin-Hong;Nam, Yun-Gyu;Ju, Jung-Il;Jung, Jae-Hong;Lee, Jong-Soo;Kim, Ha-Kun
    • Microbiology and Biotechnology Letters
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    • v.40 no.2
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    • pp.111-116
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    • 2012
  • In this study, we investigated the microbial flora changes in Gugija-Liriope tuber Makgeolli during fermentation and storage periods. We brewed Gugija-Liriope tuber Makgeolli for a week through twostage fermentations and stored the fermentation broth for a month at $4^{\circ}C$ or $20^{\circ}C$. We collected the samples periodically and analyzed microbial flora changes using viable cell counts and PCR-denaturing gradient gel electrophoresis (DGGE). Yeast viable cells were seen to have decreased to 13% of pre-storage levels after storage for 15 days at $20^{\circ}C$; however significant changes were not observed during storage at $4^{\circ}C$. Prolongation of storage time dramatically decreased the availability of viable cells. Yeast viable cell numbers had decreased to 38% of pre-storage levels at $4^{\circ}C$ and 4.8% at $20^{\circ}C$ after storage for 30 days. The results of the DGGE profile for yeast showed that Saccharomyces cerevisiae and Saccharomyces sp. were the predominant strains at the beginning of fermentation and throughout the whole period of storage. Viable cell counts for total bacteria had decreased to 36% of pre-storage levels after storage for 15 days but did not significantly change for the full 30 days of storage at $4^{\circ}C$. Similarly, viable cell counts for bacteria had decreased to 5% while viable cell numbers did not significantly change for the full 30 days at $20^{\circ}C$. Viable cell counts for lactic acid bacteria were performed and the results were similar to those for total bacteria. The results of the DGGE profile for bacteria showed that Weissella cibaria was the predominant strain at the beginning of fermentation. However it had disappeared by the end of fermentation, and Lactobacillus fermentum and Pediococcus acidilactici became the predominant species during storage.

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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    • 2015.05a
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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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