수종의 식물병원균(흰비단병균$\cdot$균핵병균 및 좀검은 균핵병균)이 생산하는 가수분해효소의 활성

조백호,김기청,Cho B. H.,Kim K. 한국응용곤충학회 1977 한국식물보호학회지 Vol.16 No.4

Activities of various hydrolytic enzymes produced by three plant pathogenic fungi, Sclerotium rolfsii Sacc., Sclerotinia sclerotiorum (Lieb.) deBary and Helminthosporium sigmoideum var. irregulare Crallery et Tullius, were measured. Activties and amounts of the enzymes in mycelia, cultural filtrates, and sclerotia(except of sclerotia of H. sigmoideum var. irregulare) were estimated at various pH levels in order to find out optimal pH for their enzymatic activities. Enzymes such as cellulase (ex), invertase, xylanase, $\beta-amylase$, polymethylgalacturonase, polygalacturonase, phosphatase and protease were estimated. Culture solution for production of enzymes was prepared by adding of 10g, D-glucose, 1.3g $NH_4NO_3,\; 0.5g\; MgSO_4,\;7H_2O,\; and\; 1.0g\; KH_2PO_4$ into 1 liter of potato decoction plus 2ml of micro element solution consisting of 0.2mg. Fe, 0.2mg Zn, and 0.1mg Mn as the sulphates into 1 liter of distilled water. All tested mycelia and cultural filtrates were obtained from the cultures incubarted in previous solution for ten days at $25^{\circ}C$, and sclerotia were harvested from PDA plates of 3. days old, The crude enzyme solutions were prepared according to the method of Miyazaki etal. Ten days after incubation, activities of Cx produced by Scl. sclerotiorum were higher than those of the other fung and each of Cx from three fungi showed different pH optima, such as S. rolfsii and Scl. schlerotiorum in acid side (around pH 3.0), H. sigmoideum var. irregulare in neutral side (around pH 6.3). Invertase activities of S. rolfsii were 20 times higher than those of the other fungi in all samples. All tested fungi, however, showed no significant difference between the enzymatic activities of their cultural filtrate and mycelia and the activities in sclerotia of S. rolfsii and Scl. sclerotiorum were hardly recognized. There were multiple peaks on the xylanase activity curves of three fungi in terms of pH values. High activities of the xylanase were revealed in sclerotia of S. rolfsii and Scl. sclerotiorum, and in mycelia of H. sigmoideum var. irregulare. The highest activities of $\beta-amylase$ were shown both in mycelia and cultural filtrate of H. sigmoideum var. irregulae among the tested fungi, and their optimal pH was 6.2 in both mycelia and cultural filtrate. In the S. rofsii and Sel. sclerotiorum, however, the activities of cultural filtrates were higher than those of the other fungi, and optimal pH was 3.0 and 6.2 for cultural filtrate and both mycelia and sclerotia, respectively. Activities of PMG were high in cultural filtrates of all tested fungi, especially in Scl. sclerotiorum and H. sigmoideum var. irregulare. Mycelia of themalso showed the considerable activities. Optimal pH for enzymatic activities were variable with thekind of fungi or with the samples measured. The highest activities of PG were presented by mycelia of S. rolfsii and Scl. sclerotiorum. $9.l\mu /min.\; and\; 9.5\mu g/min.$, respectively. Optimal pH for activity of PG in mycelia was around 4.5 in S. rolfsii and around 3.0 in Scl. sclerotiorum. Phosphatase of S. rolfsii and Scl. sclerotiorum was more active in acid side (optimal PH3. 5) and that of H. sigmoideum var. irregulare showed one peak each in acid, neutral and alkaline side. But the highest peak was at pH 9.5. Protease of all tested fungi was more active at pH 10.0, especially that of the cultural filtrate of H. sigmoideum var. irregualre. 수종의 식물병원균(흰비단병균, 균핵병균, 좀검은균 핵병균)이 생산하는 가수분해산소(Cellulase Cx, Invertase, $\beta-amylase$, Xylanase, PMG, PG, Phosphatase, Protease)를 균계체내의 효소 및 배양여액내의 효소와 균핵내의 효소(좀검은균핵병균은 제외)로 나누어 그의 생산량과 pH에 따른 활성의 변화를 검토한 결과를 요약하면 다음과 같다. 1. 배양 10일후의 Cx활성은 균핵병균이 다른 균에 비하여 활성이 높았고 흰비단병균과 균핵병균은 산성측(pH 3.0 부근)에서, 좀검은균핵병균은 중성측(pH6.0 부근)에서 활성이 높았다. 2. 흰비단병균의 Invertase는 다른균에 비하여 약 20배정도 높은 활성을 보였고 3균주 모드 배양여액과 균계체간에 효소활성의 차가 인정되지 않았다. 3. Xylanase의 활성은 3균주 모두 균계체, 균핵 및 배양여액에 따라 또 pH에 따라 아주 다양한 변화를 나타냈고 균핵내에서 활성이 높았다. 4. $\beta-amylase$의 활성은 공시균중 좀검은균핵병균의 균계체, 배양여액이 가장 높았다. (약 12.0ug/min) 흰비단병균과 균핵병균ㅇ서는 균계체나 균핵에서 보다 배양여액에서 높았는데 활성최적pH는 균계체, 균핵 모두 pH 6.2였으나 배양여액에서는 흰비단 병균과 균핵병균이 pH3.0이었는데 반해 좀검은균핵병균은 pH6.2였다. 5. PMG의 활성은 공시균 모두 배양여액에서 높았고 균계체에서는 균핵병균과 좀검은균핵병균이 높았으며 활성최적 pH는 균에 따라 또는 측정부분에 따라 다양하게 나타났다. 6. PG의 활성은 흰비단 병균과 균핵병균의 균계체에서 각각 9,1ug/min. 9.5ug/min으로서 가장 높았고 활성최적 pH는 흰비단병균이 pH4.5부근 균핵병균이 pH.3.0부근이었다. 7. 흰비단병균과 균핵병균의 Phosphatase는 산성측(최적 pH3.5)에서 활성이 높았고 좀검은 균핵 병균은 산성, 중성, 알카리측에서 각각 Peak가 나타났으나 최적 pH는 9.5였다. 8. 공시균주 모두 Protease는 pH 10.0에서 최고활성을 나타냈고 특히 좀검은균핵 병균의 배양여액내 효소활성이 높았다.

잎 절편의 재분화에 의한 참박 형질전환

조송미(Cho, Song-Mi),문선진(Moon, Sun-Jin),정수진(Chung, Soo-Jin),김미성(Kim, Mi-Seong),김영철(Kim, Young-Cheol),양광열(Yang, Kwang-Yeol),최용수(Choi, Yong-Soo),Sapkota, Kumar,조백호(Cho, Baik-Ho),김광상(Kim, Kwang-Sang) 한국자원식물학회 2006 한국자원식물학회지 Vol.19 No.5

수박의 대목으로 사용되고 있는 참박의 재분화조건을 확립함으로서 병저항성 형질전환 식물체를 얻어 보고자 본 실험을 실시하였다. 참박의 자엽을 잘라 재분화에 미치는 식물생장조절물질의 효과를 조사하였고 오이 galactinol synthase (CsGolS1) 유전자를 참박에 형질전환하였던 결과는 다음과 같다. 참박 신초의 재분화는 7일째 된 유묘의 자엽부위 중 전반부위에서 기장 높은 효율을 나타내었고, cytokinin으로 BA나 zeatin을 단용처리하는 것보다 옥신으로 IAA를 혼용처리하는 것이 신초 분화에 더 효과적이었다. 복합스트레스 내성 유전자인 오이 CsGolS1유전자를 pBI121 binary vector에 재조합하여 아그로박테리움을 이용, 참박에 형질전환하였던 결과, 형질전환체는 kanamycin이 첨가된 배지에서 생장하였고 PCR 및 Southern blot 분석에 의해 유식물체의 20% 정도가 형질전환체임을 확인할 수 있었다. In order to develop a disease-resistant root stock for the growth of watermelon, an efficient regeneration system of the gourd(Lagenaria leucantha Duch.) inbred line GO701-2 via organogenesis was established in this experiment. Using proximal parts of cotyledon explant excised from germinated seedling in vitro, maximum adventitious shoot formation (39%) was achieved on MS medium where cytokinin (BA) and auxin (IAA) were added at a concentration of 3mg/L and 0.1mg/L, respectively. Roots of the elongated shoots were successfully formed on MS medium without adding any plant growth regulators. The cucumber CsGolS1 gene known as a resistance gene against biotic and abiotic stresses, was constructed into the binary vector pBI121 under the control of CaMV 35S promoter. When the gene was introduced into the genome of gourd by Agrobacterium-mediated transformation, putative transgenic plants were obtained with the transformation efficiency of approximately 20 percent.

Enhanced Expression of a Gene Encoding a Nucleoside Diphosphate Kinase 1 (OsNDPK1) in Rice Plants upon Infection with Bacterial Pathogens

Song Mi Cho,조백호,Seo Ho Shin,Kwang Sang Kim,김영철,은무영 한국분자세포생물학회 2004 Molecules and cells Vol.18 No.3

A cDNA library was constructed using mRNA extracted from rice leaves infected with Xanthomonas oryzae pv. oryzae (Xoo), a bacterial leaf blight pathogen, to isolate rice genes induced by Xoo infection. Subtractive hybridization and differential screening of the cDNA library led to the isolation of many induced genes including a nucleotide diphosphate kinase 1 (OsNDPK1) and a pathogenesis-related protein 1 (OsPR1) cDNA. Nucleoside diphosphate kinases (NDPKs) are key metabolic enzymes that maintain the balance between cellular ATP and other nucleoside triphosphates (NTPs). Three other OsNDPK genes (NP922751, OsNDPK2 and OsNDPK3) found in databases were obtained by RTPCR. Three different programs for predicting subcellular targeting indicated that OsNDPK1 and NP922751 were non-organellar, OsNDPK2 plastidic, and OsNDPK3 mitochondrial. Only transcripts of OsNDPK1 accumulated strongly after infection with Xoo. When rice plants were infected with Burkholderia glumae, a bacterial grain/seedling rot pathogen, the pattern of expression of the rice NDPK genes was similar to that following infection with Xoo. OsNDPK1 gene expression was also strongly induced in response to exposure to salicylic acid, jasmonic acid, and abscisic acid, although the level of transcripts and their pattern of expression depended on the inducer.

Induced systemic resistance and plant growth promotion of a phosphate-solubilizing bacterium, Enterobactor intermedium 60-2G

김영철,김철홍,김길용,조백호,Kim, Young-Cheol,Kim, Chul-Hong,Kim, Kil-Young,Cho, Baik-Ho Korean Society of Soil Science and Fertilizer 2002 한국토양비료학회지 Vol.35 No.4

인산가용 미생물인 Enterobacter intermedium 60-2G의 식물생장촉진 능력과 흑성병균인 Cladosporium cucumerinum에 대한 유도전신저항성 능력을 오이와 오이 흑성병균을 모델로 확인하였다. E. intermedium 을 처리한 오이는 물을 처리한 control에 비해 C. cucumerinum에 의한 흑성병의 병징이 현저히 감소하였다. 또한 E. intermedium를 처리한 오이는 식물 생장촉진효과도 보였다. Strain 60-2G은 Fusarium oxysporum와 Magnaporthe grisea을 포함한 여러 식물 병원 곰팡이에 대해 강한 항균활성을 나타내었다. 본 연구는 인산가용능력을 가진 E. intermedium 60-2G는 식물의 생육을 증진시키는데 관여하는 여러가지 유익한 형질을 가진 아주 유용한 생물적 방제균임을 밝혔다. A phosphate-solubilizing bacterium, Enterobacter intermedium 60-2G, was examined for plant growth-promotion and induction of systemic resistance using a model system of cucumber and scab disease, caused by Cladosporium cucumerinum. Compared with a nonbacterized control, treatment of cucumber with E. intermedium significantly reduced the severity of scab disease after challenge-inoculation with C. cucumerinum. Treatment of cucumber with E. intermedium also enhanced cucumber growth. The 60-2G strain showed a strong antimicrobial activity against several plant pathogenic fungi including Fusarium soysporum and Magnaporthe grisea. These results suggest the E. intermedium 60-2G is a promising candidate as a biological control agent displaying multiple beneficial properties to promote plant health.

Pestalotiopsis menezesiana에 의한 참다래 잎마름병과 발생생태

박지영,이웅,송동업,성기영,조백호,김기청,Park, Ji-Young,Lee, Woong,Song, Dong-Up,Seong, Ki-Young,Cho, Baik-Ho,Kim, Ki-Chang 한국식물병리학회 1997 Plant Pathology Journal Vol.13 No.1

Etiological and ecopathological studi88888es on the leaf blight of kiwifruit (Chinese gooseberry), a disease newly found in 1993 in Korea, were carried out in this experiment. The leaf blight symptom was prevalent in the southern area of Chonnam Province where kiwifruits were widely cultivated. The disease occurred from July to fall, causing early defoliation of kiwifruit vines. Small brownish circular or irregular lesions appeared initially on the leaves, starting from the marginal leaf areas, and enlarged. Afterwards small grayish black granules (acervuli) were formed on the lesions. The color of old lesions changed to grayish brown or grayish white. The causal fungus was identified as Pestalotiopsis menezesiana (Bress. & Torr.) Bissett according to its morphological characters and pathogenicity (pathogenic to Chinese gooseberry and grapevine, but not to other hosts of Pestalotiopsis spp.such as tea, apple, persimmon, and chestnut). As P. longiseta and P. neglecta were reported recently in Japan as pathogens causing similar blight symptoms in kiwifruit, P. menezesiana is a new leaf blight pathogen in kiwifruit. Development of the disease was stimulated by wounding, prevailing wind, and drought. The causal fungus was proven to be overwintered in the diseased leaves on the ground surface.

빙핵활성 세균으로부터 빙핵활성 단백질의 정제 및 특성

김기청,이웅,송동업,조백호,Kim, Ki-Chung,Lee, Ung,Song, Dong-Up,Cho, Baik-Ho 한국식물병리학회 1996 Plant Pathology Journal Vol.12 No.1

3종의 빙핵세균 Peudomonas syringae 8401, Pseudomonas fuorescens 8701, Erwinia herbicola 8701의 세포 외막으로부터 아무런 변성제도 사용치 않고 sucrose density gradient centrifugation, Sephacryl gel filtration chromatography, DEAE-cellulose ion exchange chromatography, non-denaturing buffer를 이용한 PAGE, electroelution, SDS-PAGE를 통해 빙핵활성 단백질을 고도로 정제할 수 있었다. P. suringae와 P. fluorescens에서는 각각 3종류(155 kD, 75 kD, 50 kD)의 빙핵활성 단백질이, E. herbicola에서는 155 kD를 제외한 2종류(75 kD, 50 kD)의 빙핵활성 단백질은 이 연구를 통해 처음 밝혀진 것으로 , 지금까지 보고된 빙핵활성 단백질(150 KD 이상)보다는 훨씬 작은 것이다. 이는 빙핵활성을 나타내는 단백질의 기본단위는 이 실험의 결과만에 의하면 최대 50 kD임을 시사한다. 이들 단백질은 그 유래된 세균의 종류나 또는 단백질 분자량의 크기에 관계없이 모두 -5.5~7.5$^{\circ}C$에서 물을 동결시키는 높은 빙핵활성을 갖고 있었다. 이는 지금까지 보고된 어느 정제단백질의 빙핵활성보다 높은 것이다. 정제된 단백질의 빙핵활성은 trypsin 처리에 의해 상실되었고, pH6~8범위에서는 안정하였으며, pH5이하, pH9이상에서는 활성을 상실하였다. 보존온도에 대한 영향은 3$0^{\circ}C$이상이 되면 점차 활성이 감소하는 경향을 보이다 37$^{\circ}C$이상에서는 활성이 완전히 상실되었다. 금속이온으로서 Hg\ulcorner이온과 SDS에 의해 활성이 상실되었으나 phosphatidylinositol의 첨가에 의해서는 활성이 약간 증가(-1$^{\circ}C$)하였다.

빙핵세균의 검출을 위한 특이적 항혈청 개발

이웅,권미경,성기영,조백호,김기청,Lee, Ung,Kwon, Mi-Kyung,Seong, Ki-Young,Cho, Baik-Ho,Kim, Ki-Chung 한국식물병리학회 1999 식물병연구 Vol.5 No.1

Frost injury of crops is closely related to the epiphytic population dynamics of ice nucleation-active (INA) bacteria, and the injury can be reduced by decreasing the INA bacterial population. In order to predict the epiphytic population of INA bacteria on crops, a rapid and accurate detection method has to be developed. In the previous report, we produced some antibodies against INA proteins purified from the outer membrane of INA bacteria. However it was difficult to produce the antibodies because the purification procedures of the INA proteins were complicated, and the final yield was too low. We designed a specific peptide from the N-terminal region of INA protein by computer analysis and synthesized the peptide in vitro in this experiment. The peptide sequence was Asp-Ser-Por-Leu-Ser-Leu-His-Ala-Asp, that is corresponding to the highly conserved region in several INA proteins, with predicted beta turn, coiling, and hydrophilic region. A polyclonal anti-INA peptide antiserum produced specifically recognized INA bacteria as few as 10 colony-forming units (CFU) in the ELISA reactions and did not respond to other non-INA bacteria. Serological specificity of the anti-INA peptide antiserum will facilitate the forecasting of the INA bacterial population dynamics on crops.

벼(Oryza sativa L.) 내염성 품종과 감수성 품종의 유묘 염 처리 시 아미노산 및 단백질 함량 차이

신서호(Seo Ho Shin),이영만(Young Man Lee),조백호(Baik Ho Cho) 한국육종학회 2004 한국육종학회지 Vol.36 No.5

The changes of the amino acid content including proline and protein content in response to NaCl treatment were investigated in between the salt-resistant variety CI5309 and salt-susceptible variety IR28 of rice. The amino acid contents such as -aminoapid

오이와 호박 뿌리의 저온 순화에 관여하는 원형질막 H⁺-ATPase의 역할

안성주(Ahn S. J.),임양주(Im Y. J.),정갑채(Chung G. C.),조백호(Cho B. H.) 한국원예학회 1998 원예과학기술지 Vol.16 No.1

단보 : 석회유황합제가 배나무 낙엽의 진균 다양성 변화에 미치는 영향

민광현 ( Kwang Hyun Min ),송장훈 ( Jang Hoon Song ),조백호 ( Baik Ho Cho ),양광열 ( Kwang Yeol Yang ) 한국균학회 2015 韓國菌學會誌 Vol.43 No.4

석회유황합제의 배나무 동계방제 수단으로써 효과를 과학적으로 증명하기 위해 배 과수원에서 1차 전염원의 역할을 하는 낙엽에 석회유황합제를 처리한 후 낙엽에 존재하는 진균의 다양성 변화를 확인하였다. 물이 처리된 대조구와 석회유황합제 처리구의 낙엽에서 진균 genomic DNA를 추출한 다음 rDNA internal transcribed spacer (ITS) 영역의 염기서열 분석을 통해 진균들을 동정한 결과, 대조구와 석회유황합제 처리구에서 자낭균문(Ascomycota)과 담자균문(Basidiomycota)에 속하는 다양한 종류의 진균들이 동정되었으나 동정되는 진균의 수와 비율은 서로 다르게 나타났다. 특히 배 과피얼룩병을 일으키는 Alternaria속과 Cladosporium속의 진균은 대조구에 비해 석회유황합제 처리구에서 동정되는 비율이 대폭 감소되었으며, Phomopsis속의 진균은 대조구에서는 동정되었으나 석회유황합제 처리구에서는 동정되지 않았다. 이러한 결과는 석회유황합제 처리가 배 낙엽에 존재하는 진균 다양성의 변화에 영향을 미쳤으며 동계방제로서 역할을 수행할 수 있음을 시사한다. This study was conducted to examine changes in the fungal community on fallen leaves of pear by treatment with lime sulfur. Although the lime sulfur could reduce the primary inoculum of several pathogens on spring season, the effect of lime sulfur has not been well determined scientifically. Fallen leaves infected by pear diseases in pear orchards in Naju were collected and treated with lime sulfur or water as a control. To determine the fungal diversity from each treatment, rDNA internal transcribed spacer (ITS) regions were analyzed after extraction of fungal genomic DNA from lime sulfur-treated or water-treated fallen leaves, respectively. The most common fungal species were Ascomycota and Basidiomycota in both treated leaves. However, the population dynamics of several fungal species including Alternari sp., Cladosporium sp., and Phomopsis sp., which are known as pear pathogens for skin sooty dapple disease, were quite different from each treated leaves. These results indicated that lime sulfur treatment led to changes of fungal communities on pear fallen leaves and could be applicable as a dormant spray.