• 제목/요약/키워드: fungal spores

검색결과 143건 처리시간 0.019초

고추 탄저병균의 포자 발아와 부착, 균사 생장에 미치는 화합물의 활성 검정법 확립 및 살균제의 효과 (Development of assay method for the activities of new compounds, and the effect of several fungicides against spore germination, adhesion, and myceial growth of Colletotrichum sp. causing red pepper anthracnose)

  • 김재정;김준태;박성우;박은숙;김흥태
    • 농약과학회지
    • /
    • 제7권3호
    • /
    • pp.159-168
    • /
    • 2003
  • 본 실험에서는 microtiter plate를 이용하는 실내 검정법을 이용하여 고추 탄저병균의 기주체 침입 초기에 볼 수 있는 형태 변화인 포자 발아 및 부착과 균사 생장에 미치는 다양한 화합물의 활성을 대량으로 검정할 수 있는 방법을 확립하였고, 기존의 몇 가지 살균제가 가지는 고추 탄저병균의 초기 형태 분화 에 대한 억제 효과를 조사하였다. 본 실내 검정법을 표준화하기 위하여 MTT와 propanol 처리, 접종하는 포자의 농도, 배양 시간들이 병원균의 흡광도에 미치는 영향을 조사하였다. MTT를 12 시간, propanol을 1 시간 처리하였을 때, 그리고 JC24 의 포자를 $1\times10^5$개/mL의 농도로 접종하고 36 시간 배양하였을 때, 병원균의 흡광도가 일관성 있고 높게 나타났다. 살균제의 작용 특성을 알아보기 위해서 6종의 보호용 살균제, 6종의 스테롤 생합성 저해 살균제, 1종의 호흡저해제를 선발하여 microtiter plate 대량 검정 방법에서 효과를 조사하였다. 보호용 살균제 중에서 mancozeb, chlorothalonil, dithianone 등은 $6.25{\mu}g/mL$ 처리구에서도 포자의 발아, 부착과 균사의 생장을 모두 80% 이상 억제하였으나, propineb, iminoctadine, fluazinam 등은 $100{\mu}g/mL$의 처리를 2 시간만에 세척하였을 때 흡광도가 감소하는 것으로 보아 포자 부착 억제 효과는 적었다. 6종의 스테롤 생합성 저해제 중에서 hexaconazole, tebuconazole, myclobutanil 등은 포자의 발아와 균사 생육을 억제하고, 세척할 경우 JC24의 생육 억제 효과가 떨어지는 것을 보아 부착 억제 효과는 기대할 수 없었다. Nuarimol 은 $100{\mu}g/mL$로 처리한 고농도에서만 부착에 대한 억제 효과가 54.7%로 다른 억제 효과보다 우수하였으나, $6.25{\mu}g/mL$의 저농도로 처리하였을 때는 부착 억제 효과가 나타나지 않았다. Kresoxim-methyl은 6.25와 $100{\mu}g/mL$의 농도에서 포자 부착에 대한 억제율이 27.3%와 54.4%로 나타났는데, 처리하는 농도가 높아짐에 따라서 효과도 증가하였다. 이상의 결과와 같이 때 microtiter plate를 사용하여 병원균이 식물체를 침입하는 초기에 나타나는 포자 발아와 부착, 그리고 균사 생장을 검정할 수 있었으며, 본 실험 방법을 통하여 Kresoxim-methyl은 탄저병균의 포자 부착을 특이적으로 억제하는 효과가 있음을 알 수 있었다.

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

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
    • /
    • 한국균학회 2015년도 춘계학술대회 및 임시총회
    • /
    • pp.45-46
    • /
    • 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%.

  • PDF

토양조건(土壤條件)에 따른 수종(數種) 진균포자(眞菌胞子)의 발아(發芽) (Germination of Some Fungal Spores under Different Soil Conditions)

  • 심재욱;이민웅
    • 한국균학회지
    • /
    • 제16권1호
    • /
    • pp.41-48
    • /
    • 1988
  • 경기도내(京畿道內) 12개 지역(地域)에서 채집(採集)된 102개 토양(土壤)을 대상으로 하여 토양(土壤)pH, 토양수분함량(土壤水分含量), 농작물(農作物)의 경작(耕作)등 토양(土壤)의 상태에 따라 Fusarium종(種)의 포자발아율(胞子發芽率)을 조사하고 포자발아(胞子發芽)에 대한 유발토양(誘發土壤)과 억제토양(抑制土壤)의 토양미생물(土壤微生物) 분포수(分布數)를 조사하였다. 그 결과는 다음과 같다. 1. Fusarium solani f. sp. pisi는 98개 토양에서 Fusarium oxysporum f. sp.cucumerium은 95개 토양에서 Fusarium oxysporum f. sp. raphani는 82개 토양에서 Fusarium coccophilum, Fusarium coccophilum은 83개 토양에서 각각 50% 미만의 포자발아율(胞子發芽率)를 나타내었다. 2. 토양pH를 4개 군(群)으로 나눈 후 조사하였을 때 Fusarium solani f. sp. pisi, Fusarium oxysporum f. sp. cucumerium은 pH 5.1-6.0의 21개 토양에서 Fusarium oxysporum f. sp. raphani는 pH4.1-5.0의 33개 토양에서 포자발아율(胞子發芽率)이 가장 높았다. 3. 토양수분함량을 5개 군(群)으로 나눈 후 조사하였을 때 Fusarium solani f. sp. pisi, Fusarium coccophilum, Fusarium oxysporum f. sp. cucumerium은 41-60%의 토양수분함량(土壤水分含量)을 지닌 4개 토양에서 Fusarium f. sp. raphani는 토양수분함량(土壤水分含量) 21-30%의 22개 토양에서 가장 높은 포자발아율(胞子發芽率)을 나타내었다. 4. 토양(土壤)pH와 포자발아(胞子發芽), 토양수분함량(土壤水分含量)과 포자발아(胞子發芽)에 대한 상관관계(相關關係)의 조사에 있어서 토양(土壤)pH와 포자발아율(胞子發芽率)에서는 상관관계(相關關係)가 없었고 Fusarium oxysporum f. sp. raphani에서만 토양수분함량(土壤水分含量)과 포자발아율(胞子發芽率)사이에 고도의 정(正)의 상관관계(相關關係)(r=0.29)가 있었다. 5, 농작물(農作物) 경작지(耕作地) 토양(土壤)과 비경작지(非耕作地) 토양(土壤)에서 각Fusarium종(種)의 포자발아(胞子發芽)은 모두 고도의 통계적(統計的)인 유의성(有意性)이 있었으며 두 대조토양 사이에서는 오직 Fusarium solani f. sp. pisi만 통계적(統計的)인 유의성(有意性)이 인정되었다. 6. 포자발아(胞子發芽)에 대한 억제토양(抑制土壤)과 유발토양(誘發土壤)의 미생물분포수(微生物分布數)는 고선란(故線蘭), 세균(細菌), 곰팡이의 순서로 많았으며 억제토양(抑制土壤)이 유발토양(誘發土壤)보다 미생물분포수(微生物分布數)가 훨씬 높았으며 억제토양(抑制土壤)과 유발토양(誘發土壤)에서 미생물(微生物)의 분포수(分布數)는 유발토양(誘發土壤)의 세균분포수(細菌分布數)를 제외하고 모두통계적(統計的)인 유의성(有意性)이 인정되었다.

  • PDF