토양(土壞)에서 분리(分離)한 Chitinase 생성(生成) 방선균(放線菌)의 분류동정(分類同定)

김영일,김용웅,김광식,Kim, Yeong-Yil,Kim, Yong-Woong,Kim, Kwang-Sick 한국토양비료학회 1988 한국토양비료학회지 Vol.21 No.3

Four actinomycetes which produces extracellular chitinase were isolated from soil and organic matter all over the Chonnam provincial area. The chemical composition, morphological, cultural and physiological properties of isolated strain S-25, S-42, S-172 and S-267 were studied in relation to the toxonomical properties. All of strains contained phospholipids such as phosphatidyl ethanolamine, phosphatidyl inositol and diphosphatidyl glycerol. The components of cell wall in all strains have L, L-DAP, Glutamic acid, Alanine, Glycine and Glucosamine. The surface of spore is smooth and colony is grey in all strains, Based on the results obtained in these experiments all of strains are identified as Streptomyces sp.S-25, Streptomyces sp.S-42, Streptomyces sp.S-l72 and Streptomyces sp. S-267. 전남일원(全南一圓)의 토양(土壤) 및 유기물(有機物)로부터 분리(分離)된 세포외(細胞外) Chitinase활성(活性)이 높은 방선균(放線菌) S-25, S-42, S-172, S-267균주(菌株)의 분류학적(分類學的) 특성(特性)을 검토(檢討)한 결과(結果)는 다음과 같다. 1. 형태적(形態的)으로 전균주(全菌株)가 Smooth한 표면구조(表面構造)의 포자(胞子)와 회색(灰色)의 기균사를 형성(形成)하였다. 그러나 포자(胞子)사슬의 형태(形態)는 S-25, S-172균주(菌株)가 Spiral type, S-42균주(菌株)는 Recticulum apertum type이었으며 S-267균주(菌株)는 Rectiflexibles type이었다. 2. 균주(菌株)의 화학적(化學的) 조성(組成)은 세포벽(細胞壁) 구성성분(構成成分)으로 L.L-diamininopimeric acid와 Glycine을 Phospholipid로서 Phosphatidyl inositol과 Phosphatidyl ethanolamine을 함유(含有)하였다. 3. 생리적(生理的) 특징(特徵)으로써 전균주(全菌株)가 호기성(好氣性)이었으며, 탄소원(炭素源)으로 Sucrose를 제외(除外)한 대부분의 당(糖)과 유기물(有機物)을 이용(利用)하였다. 4. 이상의 분류학적(分類學的) 특성(特性)을 검토(檢討)한 결과(結果) 각(各) 균주(菌株)를 Streptomyces sp. S-24, Streptomyces sp. S-42, Streptomyces sp. S-172 그리고 Streptomyces sp. S-267로 분류(分類), 동정(同定)하였다.

土壤에서 分離한 S treptomyces Sp. s - 45 의 醒素學的 性質에 관한 硏究

金永日(Yeong-Yil Kim),金容雄(Yong-Woong Kim),金廣植(Kwang-Sik Kim) 한국토양비료학회 1988 한국토양비료학회지 Vol.21 No.2

土壞에서 分離한 Chitinase 生成 放線菌의 分類同定

金永日(Yeong-Yil Kim),金容雄(Yong-Woong Kim),金廣植(Kwang-Sick Kim) 한국토양비료학회 1988 한국토양비료학회지 Vol.21 No.3

식물(植物) 병원성(病源性) 미생물(微生物)에 미치는 Pseudomonas fluorescens, KR-164의 영향(影響)

이영환,김영일,이재평,김용웅,김용재,이재와,Rhee, Young-Hwan,Kim, Yeong-Yil,Lee, Jae-Pyeong,Kim, Yong-Wong,Kim, Yong-Jae,Lee, Jae-Wha 한국토양비료학회 1990 한국토양비료학회지 Vol.23 No.1

The antagonistic fluorescent pseudomonas, which was isolated from continuous cropping rhizosphere of pepper and cucumber, was identified as Pseudomonas fluorescens (P.f.). For further study, transformant was derived from the isolated P.f. after spontaneous mutation to give antibiotic resistance to nalidixic acid and rifampicin as marked strain. Both P.f. and transformant strains were used for this study and the results obtained were summarized as follows. 1. One of the most effective antagonistic strain, KR164, was selected against F. solani, F. oxysporum, R. solani and this strain was identified and classified as Pseudomonas fluorescens biotype IV. 2. Transformant, KR1641, was derived from strain KR164 and both strains had the same biological and biochemical characteristics. 3, Mycelial lysis and abnormal mycelia of plant pathogenic fungi were microscopically observed after simultaneous culture of fungus and given bacterial strain. 4. The length of chinese cabbage to the autolyzed became longer with given bacterial strain in dark culture. 5. Percentage of germination, number of leaves, length of height, and length of root in chinese cabbage in pot experiment were improved by inoculation of given bacterial strain. 6. The number of given bacterial strain kept generally stable until 34 days after inoculation of itself in pot experiment. Inoculation of given bacterial strain did affect the number of plant disease fungi to be decreased but did not affect the number of other bacteria, Bacillus, in pot experiment. 고추, 오이 등의 시설원예(施設園藝) 연작토양(連作土壤)에서 토양전염성(土壤傳染性) 식물병원균(植物病院菌)에 길항력(拮抗力)을 갖는 Pseudomonas fluorescens를 분리(分離), 동정(同定)하고 이 균주(菌株)를 nalidixic acid와 rifampicin에 내성을 갖는 spontaneus multant를 유도(誘導)하여 표식균주(標識菌株)를 만들었다. 이들 표식균주(標識菌株)와 모균주(母菌株)의 생화학적 특성(特性), 병원성 사장균(絲狀菌)에 대한 길항력(拮抗力), 암소배양(暗所培養), 작물(作物) pot 실험(實驗) 결과(結果)를 요약(要約)하면 다음과 같다. 1. 고추, 오이 등의 시설원예(施設園藝) 연작지(連作地) 토양(土壤)에서 형광성(螢光性) Pseudomonas를 분리(分離)하여 Rhizoctonia solani, Fusrium oxysporum, Fusarium solani에 생육(生育) 저해력(沮害力)이 우수한 KR-164를 선발(選拔)하고 이들의 분류학적(分類學的) 성질(性質)을 검토(檢討)한 결과(結果) Pseudomonas fluorescens biotype IV로 분류(分類), 동정(同定)하였다. 2. KR-164에서 표식균주(標識菌株)인 KR-1641 변이주(變異株)를 유도하였고, 이들 표식균주(標識菌株)와 모균주(母菌株)의 생화학적(生化學的) 성질(性質)은 동일(同一)하였다. 3. 분리균주(分離菌株)와 식물(植物) 병원성 사장균(絲狀菌)을 대치배양(對置培養)하여 검경한 결과(結果) 여러 형태(形態)의 균사이장(異狀)이 관찰(觀察)되었다. 4. 분리균주(分離菌株)를 배추 암소배양(暗所培養)에 처리(處理)한 결과(結果) 무처리구(無處理區)에 비해 생육기간(生育其間)이 현저하게 연장(延長) 되었다. 5. 분리균주(分離菌株)의 접종(接種)이 대조구(對照區)와 사장균(絲狀菌) 접종구(接種區)에 비해 배추의 발아율(發芽率) 및 엽수, 초장, 근장 등의 생육(生育)을 향상(向上)시켰다. 6. 토양(土壤)에 접종(接種)된 표식균주(標識菌株)는 접종(接種) 35일까지 안정(安定)하였으나 병원성 사장균(絲狀菌)은 현저하게 감소(減少)하였고, 표식균주(標識菌株)는 근권(根圈) 유용미생물(有用微生物)의 생육(生育)에 뚜렷한 영향(影響)을 미치지 않았다.

Bacillus subtilis 접종이 배추 및 참깨의 生長과 土壤 微生物相에 미치는 影響

金廣植(Kwang-Sik Kim),李載平(Jae-Pyeong Lee),金容雄(Yong-Woong Kim),李英煥(Young-Hwan Rhee),金永日(Yeong-Yil Kim) 한국토양비료학회 1993 한국토양비료학회지 Vol.26 No.4

근권 길항세균 Pseudomonas fluorescens KR164 에 Bacillus thuringiensis HD - 1 유전자의 삽입과 발현

김영일,이영환,강흔수 한국농화학회 1992 Applied Biological Chemistry (Appl Biol Chem) Vol.35 No.4

The plasmids pSUPBT and pSUPBTR were constructed with a vector pSUP2021 and the BT toxin gene in the plasmid pES 1. The plasmids constructed were introduced into the antagonistic rhizobacteria P. fluorescens KR164 by conjugation and P. fluoyescens having pSUPBT and pSUPBTR were named P. fluorescens KR164(pSUPBT) # 2, KR164(pSUPBT) # 3, KR164(pSUPBTR) # 2 and KR164(pSUPBTR) # 3, respectively. The BT toxin gene were identified in all transformants by Southern hybridization and the final product of BT toxin gene was identified only in P. fluorescens KR164(pSUPBTR) # 3 by SDS-PAGE. This crystal toxin protein were also observed in electron microscopy.

식물 생장촉진 미생물의 외부 유전자 도입과 그 접종효과

김광식,김용웅,김영일,이영환 한국농화학회 1992 Applied Biological Chemistry (Appl Biol Chem) Vol.35 No.5

The antagonistic bacteria, showing distinguished effect against Fusarium oxysporum and Rhizoctonia solani were isolated from the rhizosphares of horticultural plants and identified as Bacillus subtilis. The strains were studied for their characteristics of biochemistry, physiology, antagonistic effect against plant pathogenic fungi, and growth promoting effect on horticultural plants. The Bacillus thuringiensis(BT) HD-1 toxin gene was introduced into these B. subtilis. The BT toxin genes on chromosome of the bacteria were identified by southern blotting, but its proteins were not detected by SDS-PAGE. These transformed bacteria showed growth promoting effect and showed also insecticidal and antagonistic effects against Bombix mori and fungi F. oxysporum and R. solani but not against nematode Bursaphelenchus xylophilus.

식물 병원성 사상균(絲狀菌)에 결항력(拮抗力)을 갖는 Serratia marcescens CK - 3 의 분리 및 효소적 성질

김광식,김종현,전우복,김영일,이영환,박화성,이재화 한국농화학회 1991 Applied Biological Chemistry (Appl Biol Chem) Vol.34 No.1

Serratia marcescens CK-3, decomposing chitin which is a major component of cell wall in phytopathogenic fungi, was isolated from the continuous cropping rhizosphere of pepper and cucumber and its enzymatic property was examined. S. marcescens CK-3 was found to have antagonistic effects against Fusarium axysporum and Rhizoctania solani and to have complex enzyme systems such as chitinase, laminarinase, and proteinase. The preferable composition of the medium for production of chitinase was found and was as follows : colloidal chitin 1.5%, tryptone 0.5%, glucose 1.0%., peptone 0.2%, MgSO₄·7H₂O 0.1 %, K₂HPO₄ 0.1 %, and NaCl 0.1 % (w/v), pH 6.8. The maximum enzyme production was observed after culture of 72 hours at 30℃ using a medium containing the above chemical composition. The optimal pH and temperature for in vitro activity of chitinase from S. marcescens CK-3 were pH 7.5 and 50℃, respectively. The enzyme activity increased by metal ions such as Ag^+ and Mn^(++).