• Title/Summary/Keyword: Fungi

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Development of Analytical Method for Ergot Alkaloids in Foods Using Liquid Chromatoraphy-Tandem Mass Spectrometry (LC-MS/MS를 이용한 식품 중 맥각 알칼로이드 시험법 개발)

  • Chun, So Young;Chong, Euna;Lee, Bomnae;Kwon, Jin-Wook;Park, Hye Young;Kim, Sheenhee;Gang, Giljin
    • Journal of Food Hygiene and Safety
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    • v.34 no.2
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    • pp.158-169
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    • 2019
  • Ergot alkaloids are mycotoxin produced by fungi of the Claviceps genus, mainly by Claviceps purpurea in EU. Recently obtained informations indicates necessity for control the ergot in imported grains. Recent occurrence data of ergot alkaloids from EU countries indicate the necessities of management and control these toxins from the imported grains like rye, wheat, oat etc. The aim of this study is to optimize the liquid chromatography-tandem mass spectrometry method for determination of ergot alkaloids (ergometrine, ergosine, ergotamine, ergocornine, ergocryptine, ergocristine and their epimers (-inines) from grain and grain-based food. The test method was optimized by extracting the sample with acetonitrile containing 2 mM ammonium carbonate, purification with Mycosep cartridge, and instrumental analysis by LC-MS/MS using Syncronis C18 column. The standard calibration curves showed linearity with correlation coefficents; $R^2$ >0.99. Mean recoveries ranged from 72.0 to 111.3% at three different fortified levels (20, 50, and $100{\mu}g/kg$). The correlation coefficient expressed as precision was within the range of 1.9-12.9%. The limit or quantifications (LOQ) ranged from 0.012 to $0.058{\mu}g/kg$. The developed analytical method met the criteria of AOAC Int. and CAC validation parameters like accuracy and sensitivity. As a result, it was confirmed that the test method developed in this study is suitable for the simultaneous analysis of six species of ergot alkaloid from grains and grain products.

Distribution and Antimicrobial Resistance of Non-Tuberculous Mycobacteria during 2015~2020: A Single-Center Study in Incheon, South Korea (2015~2020년 동안 인천 지역 단일기관에서의 비결핵항산균 분포 및 항균제 내성률)

  • Kim, Jiwoo;Ju, Hyo-Jin;Koo, Jehyun;Lee, Hyeyoung;Park, Hyeonhwan;Song, Kyungcheol;Kim, Jayoung
    • Korean Journal of Clinical Laboratory Science
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    • v.53 no.3
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    • pp.225-232
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    • 2021
  • This study sought to investigate the distribution, antimicrobial resistance rate, and bacterial co-infection frequency of non-tuberculous mycobacteria (NTM) in a single center in Incheon, South Korea. A total of 8,258 specimens submitted for tuberculosis (TB)/NTM real-time PCR tests during the years 2015 to 2020 were retrospectively reviewed. In total, 296 specimens (3.6%) were NTM positive, and the positivity increased from 2.5% (30/1,209) in 2015 to 3.8% (66/1,740) in 2020. Of 296 NTM specimens, 54.7% (162/296) were identified as the Mycobacterium avium complex (MAC) followed by the Mycobacterium abscessus complex (MABC) 20.9% (62/296), M. fortuitum 6.4% (19/296) and M. flavescens 3.4% (10/296). Of the NTM-positive specimens, 76.7% (227/296) were tested for drug resistance. The results showed multidrug-resistant NTM in 40.1% (91/227) and extensively drug-resistant NTM in 59.9% (136/227) of these specimens. Of the 145 isolates taken for bacterial culture, bacteria/fungi co-infection with NTM accounted for 43.4% (63/145), in which the most common bacterial species was Klebsiella pneumonia (23.8%, 15/63). This study is the first report on the distribution and antimicrobial resistance of NTM in Incheon. As the proportion of NTM infections increases, active treatment and thorough infection control are required for effective management.

Application Effects of Fermented Mixed Organic Fertilizer Utilizing By-Products on Yield of Chinese Cabbage and Soil Environment (부산물 활용 발효 유기질비료가 배추 수량 및 토양환경에 미치는 영향)

  • An, Nan-Hee;Lee, Sang-Min;Oh, Eun-mi;Lee, Cho-Rong;Gong, Min-Jae
    • Journal of the Korea Organic Resources Recycling Association
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    • v.28 no.4
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    • pp.77-85
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    • 2020
  • This study analyzes the effects of mixed fermented organic fertilizer on chinese cabbage growth and soil properties in order to investigate the nutritional effects of organic fertilizers, which are developed as an alternative fertilizer for imported castor oil cake. In this study, four treatments were set up: 100% and 200% rate of nitrogen application (320 kg ha-1 for Chinese cabbage) on mixed fermented organic fertilizer A(FA) and mixed fermented organic fertilizer B(FB), respectively, 100% rates of the mixed expeller cake (MEC) fertilizer, and the untreated control. Results revealed that the growth and yield of Chinese cabbage increased as more fermented organic fertilizer was used. However, while there were no significant differences in growth characteristics between treatments of 100% rate of mixed fermented organic fertilizer and 100% rate of MEC, the impacts on yields resulted similar. The nitrogen use efficiency (NUE) of Chinese cabbage was measured a range of 20-31% depending on the response to treatment. The 100% FA showed the same as NUE and nitrogen absorption with 100% rate of MEC. Regarding soil properties after cultivation, there were no significant differences among the effects of fertilizers in pH, EC, soil organic matter, and available phosphate. However, the content of exchangeable cations(K, Ca, Mg) was higher in areas treated with mixed fermented organic fertilizer than in untreated areas. Furthermore, the bacterial population density in the soil was higher in areas treated with mixed fermented organic fertilizer than in untreated areas and increased as more mixed fermented organic fertilizer was used. There were no significant differences in the population density of actinomycetes and fungi when fertilizer was applied to the soil. These results also show that FA, as a alternative organic fertilizer for imported castor oil cake, has similar nutritional effects as that of MEC. Therefore, further research the appropriate amounts of fertilizer is required to achieve economical and eco-friendly nutrient management.

Complete Genome Sequence and Antimicrobial Activities of Bacillus velezensis MV2 Isolated from a Malva verticillate Leaf (아욱 잎에서 분리한 Bacillus velezensis MV2의 유전체 염기서열 분석과 항균활성능 연구)

  • Lee, Hyeonju;Jo, Eunhye;Kim, Jihye;Moon, Keumok;Kim, Min Ji;Shin, Jae-Ho;Cha, Jaeho
    • Microbiology and Biotechnology Letters
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    • v.49 no.1
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    • pp.111-119
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    • 2021
  • A bacterial strain isolated from a Malva verticillata leaf was identified as Bacillus velezensis MV2 based on the 16S rRNA sequencing results. Complete genome sequencing revealed that B. velezensis MV2 possessed a single 4,191,702-bp contig with 45.57% GC content. Generally, Bacillus spp. are known to produce diverse antimicrobial compounds including bacteriocins, polyketides, and non-ribosomal peptides. Antimicrobial compounds in the B. velezensis MV2 were extracted from culture supernatants using hydrophobic interaction chromatography. The crude extracts showed antimicrobial activity against both gram-positive bacteria and gram-negative bacteria; however, they were more effective against gram-positive bacteria. The extracts also showed antifungal activity against phytopathogenic fungi such as Fusarium fujikuroi and F. graminearum. In time-kill assays, these antimicrobial compounds showed bactericidal activity against Bacillus cereus, used as indicator strain. To predict the type of antimicrobial compounds produced by this strain, we used the antiSMASH algorithm. Forty-seven secondary metabolites were predicted to be synthesized in MV2, and among them, fourteen were identified with a similarity of 80% or more with those previously identified. Based on the antimicrobial properties, the antimicrobial compounds may be nonribosomal peptides or polyketides. These compounds possess the potential to be used as biopesticides in the food and agricultural industry as an alternative to antibiotics.

Forecasting Leaf Mold and Gray Leaf Spot Incidence in Tomato and Fungicide Spray Scheduling (토마토 재배에서 점무늬병 및 잎곰팡이병 발생 예측 및 방제력 연구)

  • Lee, Mun Haeng
    • Journal of Bio-Environment Control
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    • v.31 no.4
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    • pp.376-383
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    • 2022
  • The current study, which consisted of two independent studies (laboratory and greenhouse), was carried out to project the hypothesis fungi-spray scheduling for leaf mold and gray leaf spot in tomato, as well as to evaluate the effect of temperature and leaf wet duration on the effectiveness of different fungicides against these diseases. In the first experiment, tomato leaves were infected with 1 × 104 conidia·mL-1 and put in a dew chamber for 0 to 18 hours at 10 to 25℃ (Fulvia fulva) and 10 to 30℃ (Stemphylium lycopersici). In farm study, tomato plants were treated for 240 hours with diluted (1,000 times) 30% trimidazole, 50% polyoxin B, and 40% iminoctadine tris (Belkut) for protection of leaf mold, and 10% etridiazole + 55% thiophanate-methyl (Gajiran), and 15% tribasic copper sulfate (Sebinna) for protection of gray leaf spot. In laboratory test, leaf condensation on the leaves of tomato plants were emerged after 9 hrs. of incubation. In conclusion, the incidence degree of leaf mold and gray leaf spot disease on tomato plants shows that it is very closely related to formation of leaf condensation, therefore the incidence of leaf mold was greater at 20 and 15℃, while 25 and 20℃ enhanced the incidence of gray leaf spot. The incidence of leaf mold and gray leaf spot developed 20 days after inoculation, and the latency period was estimated to be 14-15 days. Trihumin fungicide had the maximum effectiveness up to 168 hours of fungicides at 12 hours of wet duration in leaf mold, whereas Gajiran fungicide had the highest control (93%) against gray leaf spot up to 144 hours. All the chemicals showed an around 30-50% decrease in effectiveness after 240 hours of treatment. The model predictions in present study could be help in timely, effective and ecofriendly management of leaf mold disease in tomato.

Effect of thinning ratio on the forest environment and fruiting of ectomycorrhizal mushrooms in a Pinus densiflora stand (소나무림에서 간벌률이 산림 내 환경과 외생균근성 버섯 발생에 미치는 영향)

  • Yong-Woo Park;Jin-Gun Kim;Hwayong Lee
    • Journal of Mushroom
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    • v.21 no.1
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    • pp.22-32
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    • 2023
  • To investigate the effect of thinning intensity on environmental factors and ectomycorrhizal mushroom fruiting in forest ecosystems, we studied canopy closure, throughfall, soil temperature, soil moisture, light response of understory vegetation, and ectomycorrhizal mushroom fruiting in a 10-year-old pine forest after 34%, 45%, and 60% thinning. Canopy closure was significantly higher in the 34% treatment and control plots, ranging from 80-85% in April. However, in November, all thinning treatment plots showed a decrease of approximately 5-10% compared with the control plot. The 60% treatment plot had over 200 mm of additional throughfall compared with the control plot, and monthly throughfall was significantly higher by more than 100 mm in October. The soil temperature in each treatment plot increased significantly by up to 1℃ or more compared with the control plot as the thinning rate increased. The soil moisture increased by more than 5% in the thinning treatment plots during rainfall, particularly in the 34% treatment plot, where the rate of moisture decrease was slower. The photosynthetic rate of major tree species (excluding Pinus densiflora)was highest in Quercus mongolica, with a rate of 7 µmolCO2·m-2·s-1. At a lightintensity of 800 μmol·m-2·s-1, Q. mongolica showed the highest photosynthetic level of 6 ± 0.3 μmolCO2·m-2·s-1 in the 45% treatment. The photosynthetic rate of Fraxinus sieboldiana and Styrax japonicus increased as the thinning intensity increased. The Shannon-Wiener index of mycorrhizal mushrooms did not significantly differ among treatments, but the fresh weight of mushrooms was approximately 360-840 g higher in the 34% and 45% treatments than in the control. Additionally, the fresh weight of fungi in the 60% treatment was 860 g less than that in the control. There were more individuals of Amanita citrina in the control than in the thinning treatment, while Suillus bovinus numbers increased by more than 10 times in the 34% thinning treatment compared with the control.

Rumen Fermentation was Changed by Feed Inoculation Method in In Vitro (사료 접종 방법에 의한 in vitro 반추위 발효 상성 변화)

  • Yoo, Dae-Kyum;Moon, Joon-Beom;Kim, Han-Been;Yang, Sung-Jae;Park, Joong-Kook;Lee, Se-Young;Seo, Ja-Kyeom
    • ANNALS OF ANIMAL RESOURCE SCIENCES
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    • v.30 no.3
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    • pp.111-120
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    • 2019
  • The objective of this study was to investigate the effect of different feed inoculation method on rumen fermentation in an in vitro. Three experimental treatments were used: control (CON, direct dispersion of feed (2 g) in rumen fluid), combinations of direct dispersion (1 g) and nylon bag (DNB, pore size: 50 ㎛, 1 g), and nylon bag (NB, 2 g). An in vitro fermentation experiment was carried out using strained rumen fluid for 48 h incubation time and timothy was used as a substrate. At the end of the incubation, in vitro dry matter digestibility (IVDMD), in vitro neutral detergent fiber digestibility (IVNDFD), pH, volatile fatty acids (VFA), ammonia nitrogen (NH3-N), and microbial community were evaluated and gas production was estimated at 3, 6, 12, 24, 48 h incubation periods. Gas production was higher in CON than DNB and NB at 6 and 12 h incubation time (p<0.01). There were no differences in final gas production, pH, NH3-N concentration, total VFA production, and VFA profiles among treatments. The IVDMD was lowest in CON (p<0.01) but the IVNDFD was not differed by feed distribution methods. There were no significant differences in general bacteria and fungi. Protozoa count was highest in NB treatment among treatments (p<0.01). The abundance of cellulolytic bacteria, Ruminococcus flavefaciens and Fibrobacter succinogenes, was highest in the CON among treatments (p<0.01).

Analysis of the Reduction Effect of Combined Treatment with UV-C and Organic Acid to Reduce Aspergillus ochraceus and Rhodotorula mucilaginosa Contamination (Aspergillus ochraceus와 Rhodotorula mucilaginosa 저감을 위한 자외선과 유기산 복합처리 효과 분석)

  • Eun-Seon Lee;Jong-Hui Kim;Bu-Min Kim;Mi-Hwa Oh
    • Journal of Food Hygiene and Safety
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    • v.39 no.1
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    • pp.54-60
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    • 2024
  • This study investigated the effectiveness of using pathogens and aqueous acids to reduce the Aspergillus ochraceus and Rhodotorula mucilaginosa contamination in livestock production environments. For this study, 1 mL of each bacterial suspension (107-108 spores/mL) was inoculated on a knife surface, dried at 37℃, and used under each treatment condition. First, to investigate the effect of organic acids, acetic, lactic, and citric acids were used. Subsequently, to select the appropriate concentration, they were prepared at concentrations of 0.5, 1, 2, 3, 4, and 5%, respectively. Accordingly, to further maximize the effect of organic acid treatment, we combined the treatment with ultraviolet light. The two strains showed a significant difference (P<0.05) compared to the initial strain, with a greater than 90% decrease in the concentrations of all organic acids. Consequently, acetic and lactic acids decreased by approximately 5 and 2 log colony forming unit (CFU)/cm2, respectively, when treated with ultraviolet light (360 mJ/cm2); however, citric acid decreased by less than 1 log CFU/cm2. However, when manufactured with 4% acetic acid, a severe malodor was emitted, making it difficult for workers to use it in a production environment. Accordingly, the optimal treatment conditions for organic acid and ultraviolet light for application were selected as follows: immersion in a 4% lactic acid solution for 1 minute and then, sterilization with ultraviolet light at 360 mJ/cm2. Finally, when a pork meat sample was cut with a knife that was finally washed with lactic acid and treated with ultraviolet light, the low level of inoculum transferred from the cleaned knife to the surface of the sample was not detected. In conclusion, using this established method can prevent cross-contamination of the surface of the meat during processing.

Degradation of Poultry Feathers by Bacillus amyloliquefaciens Y10 With Plant Growth-promoting Activity and Biological Activity of Feather Hydrolyzates (식물 성장 촉진 활성을 가진 Bacillus amyloliquefaciens Y10에 의한 가금 우모의 분해 및 생산된 우모 분해산물의 생리활성)

  • Yedam Kim;Young Seok Lee;Youngsuk Kim;Jinmyeong Song;Yeongbeen Bak;Gyulim Park;O-Mi Lee;Hong-Joo Son
    • Journal of Life Science
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    • v.34 no.5
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    • pp.304-312
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    • 2024
  • This study was conducted to characterize strain Y10, isolated from discarded chicken feathers. Strain Y10 was identified as Bacillus amyloliquefaciens through phenotypic and 16S rRNA gene analysis. B. amyloliquefaciens Y10 exhibited plant growth-promoting activities, including the production of fungal cell-degrading enzymes (cellulase, lipase, protease, and pectinase), siderophores, ammonia, and indoleacetic acid. Furthermore, strain Y10 was able to inhibit the mycelial growth of several phytopathogenic fungi. When 0.1% sucrose as a carbon source and 0.05% casein as a nitrogen source were added to the basal medium, adjusted to pH 10, and cultured at 35℃, the degradation rate of chicken feathers by strain Y10 was about two times higher than that of the basal medium, with the feathers almost completely degraded in four days. Strain Y10 also degraded various keratin substrates, including duck feathers, wool, and human nails. It was confirmed that the feather hydrolyzates prepared using strain Y10 exhibited antioxidant activities, such as 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity (EC50 = 0.38 mg/ml) and superoxide dismutase-like activity (EC50 = 183.7 mg/ml). These results suggest that B. amyloliquefaciens Y10 is a potential candidate for the development of bioinoculants and feed additives applicable to the agricultural and livestock industries, as well as the microbiological treatment of keratin waste.

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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