식품병원성 곰팡이 성장을 저해하는 방선균 Streptomyces griseofuscus CNU-A91231의 분리 및 배지 조성 Isolation and Medium Development of the Actinomycetes, Streptomyces griseofuscus CNU-A91231, Inhibiting Phytopathogenic Fungi원문보기
Actinomycetes 500계통 중 식물병원성 곰팡이(Alternaria alternata, Botryotinia fuckeliana, Colletotrichum acutatum, Colletotrichum gleosporioides, Corticium sasaki, Cylindrocarpon destructans, Fusarium oxysporium, Magnaporthe grisea, Phytophthora infestans, Phytium ultimum, Thanatephorus cucumeri)저해하는 균주를 선발하였다. CNU-A91231(Korea Agricultural Culture Collection #20938)이 식물병원성 곰팡이에 대한 억제 활성이 강한 것으로 나타났으며 16s rDNA sequence를 통해 Streptomyces griseofuscus로 동정 하였다. S. griseofuscus CNUA91231의 효율적인 배양을 위해 간단하고 실용적인 배지를 개발하였다. 이 균주의 최적 성장조건은 28, pH 6에서 aeration 조건이었다. Alanine, glutamine, proline, ammonium ion은 좋은 질소원이었다. 당밀에 Na, Cl, Ca, P, K, Mg를 첨가하였을 때는 S. griseofuscus.의 성장을 증가시키지 못하였다. 아미노산을 함유한 비료를 사용하였을 때 이 균주의 성장을 증가시켰다. 최적의 배지로서 당밀 + 1% glutamate 부산물 분말로 결정하였다. 최적 배지에서 S. griseofuscus의 배양은 이 균주의 antifungal 활성에 영향을 주지 않았다. 본 연구에서 연구된 bacterium과 배지가 작물재배에 생물학적 항 곰팡이 제재로 이용될 것으로 기대된다.
Actinomycetes 500계통 중 식물병원성 곰팡이(Alternaria alternata, Botryotinia fuckeliana, Colletotrichum acutatum, Colletotrichum gleosporioides, Corticium sasaki, Cylindrocarpon destructans, Fusarium oxysporium, Magnaporthe grisea, Phytophthora infestans, Phytium ultimum, Thanatephorus cucumeri)저해하는 균주를 선발하였다. CNU-A91231(Korea Agricultural Culture Collection #20938)이 식물병원성 곰팡이에 대한 억제 활성이 강한 것으로 나타났으며 16s rDNA sequence를 통해 Streptomyces griseofuscus로 동정 하였다. S. griseofuscus CNUA91231의 효율적인 배양을 위해 간단하고 실용적인 배지를 개발하였다. 이 균주의 최적 성장조건은 28, pH 6에서 aeration 조건이었다. Alanine, glutamine, proline, ammonium ion은 좋은 질소원이었다. 당밀에 Na, Cl, Ca, P, K, Mg를 첨가하였을 때는 S. griseofuscus.의 성장을 증가시키지 못하였다. 아미노산을 함유한 비료를 사용하였을 때 이 균주의 성장을 증가시켰다. 최적의 배지로서 당밀 + 1% glutamate 부산물 분말로 결정하였다. 최적 배지에서 S. griseofuscus의 배양은 이 균주의 antifungal 활성에 영향을 주지 않았다. 본 연구에서 연구된 bacterium과 배지가 작물재배에 생물학적 항 곰팡이 제재로 이용될 것으로 기대된다.
Five hundred strains of actinomycetes were screened for inhibitory activity against the phytopathogenic fungi; Alternaria alternata, Botryotinia fuckeliana, Colletotrichum acutatum, Colletotrichum gleosporioides, Corticium sasaki, Cylindrocarpon destructans, Fusarium oxysporium, Magnaporthe grisea, ...
Five hundred strains of actinomycetes were screened for inhibitory activity against the phytopathogenic fungi; Alternaria alternata, Botryotinia fuckeliana, Colletotrichum acutatum, Colletotrichum gleosporioides, Corticium sasaki, Cylindrocarpon destructans, Fusarium oxysporium, Magnaporthe grisea, Phytophthora infestans, Phytium ultimum, and Thanatephorus cucumeris. The strain CNU-A91231 (Korea Agricultural Culture Collection #20938) showed a strong activity against the phytopathogenic fungi and it was identified as Streptomyces griseofuscus based on the sequence of 16s rDNA. Practical and simple media for the strain S. griseofuscus CNU-A91231 was developed at the conditions ($28^{\circ}C$ and pH 6 with aeration) for efficient bacterial growth. Alanine, glutamine, proline and ammonium ion were good nitrogen sources for the bacterium. Addition of the major salts including Na, Cl, Ca, P, K, and Mg into molasses did not increase the growth of S. griseofuscus. Addition of fertilizers containing amino acids significantly enhanced growth of the bacterium. The optimal medium was formulated as molasses + 1% of glutamate fermentation waste powder. All the conditions and components used in this study did not affect the antifungal activity of S. griseofuscus. The bacterium and the medium in this study can be used as a bio-antifungal agent for plant farming.
Five hundred strains of actinomycetes were screened for inhibitory activity against the phytopathogenic fungi; Alternaria alternata, Botryotinia fuckeliana, Colletotrichum acutatum, Colletotrichum gleosporioides, Corticium sasaki, Cylindrocarpon destructans, Fusarium oxysporium, Magnaporthe grisea, Phytophthora infestans, Phytium ultimum, and Thanatephorus cucumeris. The strain CNU-A91231 (Korea Agricultural Culture Collection #20938) showed a strong activity against the phytopathogenic fungi and it was identified as Streptomyces griseofuscus based on the sequence of 16s rDNA. Practical and simple media for the strain S. griseofuscus CNU-A91231 was developed at the conditions ($28^{\circ}C$ and pH 6 with aeration) for efficient bacterial growth. Alanine, glutamine, proline and ammonium ion were good nitrogen sources for the bacterium. Addition of the major salts including Na, Cl, Ca, P, K, and Mg into molasses did not increase the growth of S. griseofuscus. Addition of fertilizers containing amino acids significantly enhanced growth of the bacterium. The optimal medium was formulated as molasses + 1% of glutamate fermentation waste powder. All the conditions and components used in this study did not affect the antifungal activity of S. griseofuscus. The bacterium and the medium in this study can be used as a bio-antifungal agent for plant farming.
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제안 방법
To determine physical conditions, various sets of pH, temperature, and aeration were tested. The bacterium grew in 1% (w/v) of SBM and WML at 28℃ and 150 rpm for 3 days.
대상 데이터
The effect of nitrogen supplement on the growth of the bacterium was tested. As nitrogen sources, urea, (NH4)2SO4, KNO3, and NaNO3 were tested. Addition of urea or (NH4)2SO4 in SMB increased cell mass production of the bacterium (Fig.
Five hundred actinomycetes strains were kindly provided by the Microbial Resource Database in Korea Research Institute of Bioscience and Biotechnology (Taejeon, Korea). Phytopathogenic fungi, A.
There is a huge agricultural loss by plant pathogens: 25% in Europe, 29% in USA, 42% in Africa, and 43% in Asia [1]. In this study, biological inhibitors were screened against the filamentous phytopathogenic fungi: Anternaria alternate, Botryotinia fuckliana, Collectotrichum accutatum, Magnaporthe grisea, Phytophthora infestans, Thanatephorus cucumeris, Colletrichum gloeosporioide, Phythium ultimum, Fusarium oxysporum, Cylindrocarpon destructans, and Corticium sasaki.
grieofuscus. Panels: A, urea in SM; B, urea in WML; C, (NH4)2SO4 in SMB; D, (NH4)2SO4 in WML; E, KNO3 in SMB; F, KNO3 in% WML; G, NaNO3 with 2% SMB; H, NaNO3 with 2% WML. Symbols: ○, 0%; △, 0.
Five hundred actinomycetes strains were kindly provided by the Microbial Resource Database in Korea Research Institute of Bioscience and Biotechnology (Taejeon, Korea). Phytopathogenic fungi, A. alternata, B. fuckeliana, C. acutatum, C. gleosporioides, C. sasaki, C. destructans, F. oxysporium, M. grisea, P. infestans, P.ultimum, and T. cucumeris, were kindly provided by Chi-Hwan Lim (Dept. Environmental Chemistry, Chungnam National University, Taejeon, Korea). The fungi grew on the solid potato dextrose agar (PDA) medium (Potato Dextrose 24 g/L, Agar 15 g/L) and the actinomycetes grew in MB-7 medium (glucose, 15 g/L; peptone, 10 g/L; yeast extract, 10 g/L; and glycerol, 10 g/L).
The PDA piece (0.5×0.5 cm) covered with fungal mycelium was posed on the center of PDA plates.
griseofuscus CNUA91231. This strain was stocked in Korea Agricultural Culture Collection (Suwon, Kyungi, Korea; #20938).
성능/효과
In conclusion, the media for S. griseofuscus CNUA91231 showing a high antifungal activity against phytopathogens were developed. The optimized media could be easily prepared and used in agricultural farms.
The plates were incubated at 24℃ for a week. The effect of media and growth conditions on antifungal activity of S. griseofuscus was measured by determining the inhibition activity against growth of C. gloeosporioides (a red pepper anthracnose pathogen).
The strongest strain A91231 was further tested for classification. The full sequence of 16s rDNA in the strain A91231 blasted 100% with S. griseofuscus from GenBank database and thus the strain was partially identified and named as S. griseofuscus CNUA91231. This strain was stocked in Korea Agricultural Culture Collection (Suwon, Kyungi, Korea; #20938).
Yeast extract is a rich source for amino acids and vitamins, and thus the effect of them was tested. The preliminary experimental data showed that, out of 20 amino acids, alanine, glutamine, asparagine, and proline significantly increased the cell mass production of the bacterium. The growth by the addition of 1% (w/v) each of the amino acids with 2% molasses was measured (Fig.
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