옥수수의 바이러스병에 관한 연구 : (1)Rice Black-streaked Dwarf Virus의 形狀과 細胞內出現 (1)Morphology and Intracellular Appearance of Rice Black-streaked Dwarf Virus

張茂雄,李鍾縕 영남대학교 자원문제연구소 1987 資源問題硏究 Vol.6 No.-

Dip preparations obtained by a modified direct negative staining method revealed both complete virus particles of 80-90nm and incomplete virus particles of 7O-74nm in diameter. Complete particles correspond to the large particles that appeared in the infected cell cytoplasm outside of the viroplasm and in the vacuoles. The large particles probably represent the complete particles of rice black-streaked dwarf virus(RBSDV) and the out coat may be easily removed by phosphotungsteic acid staining. All the features associated with RBSDA infection in maize neoplastic cells, ie. viroplasm, tubular structure and virus particles, were well preserved and easily recognizable in ultrathin section preparations. Large particles 70nm in diameter were free or aligned in tubular arrangement in the cytoplasm and vacuole. All these particles had an inner darken core and an outer lighter coat(complete particles), small particles 50nm in diameter, embadded in the viroplasm, seemed to lack an outer coat (incomplete particles). In the diseased maize leaves, the virus was restricted to the vascular bundle and bundle sheath cells.

한국(韓國)에서 화훼식물(花卉植物)의 바이러스병(病) 연구현황(硏究現況)

장무웅 한국식물병리학회 1987 Plant Pathology Journal Vol.3 No.1

참깨의 모자이크 증상에서 분리한 수박모자이크 바이러스에 관한 연구

장무웅,이창은 한국응용곤충학회 1980 한국응용곤충학회지 Vol.19 No.4

참깨, 호박, 오이의 모자이크 증상주에서 전자현미경 및 즙액접종에 의하여 분리한 바이러스에 대하여 다음과 같은 결과를 얻었다. 1. 본 바이러스를 즙액접종 하였을 때 P. vutgaris, P. sativum, S. indicum, C. melo, C. maxima, C. pepo는 전신감염, C. amaranticolar는 국부감염 되었다. 2. 본 바이러스는 M. persicae로 쉽게 전반되었고, 종자전염은 하지 않았다. 9. 조즙액중의 물리성은 내열성이 (10분), 희석성이 , 내보존성이 일이었다. 4. 본 바이러스의 형태는 길이 의 사상립자이고, 본 바이러스의 감염에 의해 세포질내에서 세포질봉입체가 만들어진다는 것이 확인되었다. 5. 본 바이러스에 감염된 S. indicum, C. amaranticolor의 병엽초박절편을 전자현미경으로 관찰하였던바, 각종세포의 세포질내에 사상립자가 병행배열 또는 산재해 있고, 또 사상립자와 함께 반드시 세포질내입체(pinwheel, boundles, laminated aggregates)가 확인되었다. 6. 이상의 결과를 종합해서 참깨 모자이크병의 병원바이러스를 Watermelon mosaic virus(WMV)로 동정하였다. This paper deals with the studies on the occurence of a new virus disease of sesame and the identification of the causal virus. The virus disease of sesame has been regarded as a widespread disease in the sesame-growing areas in the southern part of Korea. The disease was found to be caused by watermelon mosaic virus (WMV). During the years since 1978, stunting of sesame plants, with yellow mosaic, necrotic spot, and malformation, were collected from 17 different places. Virus isolates from 27 out of 32 samples were identified as WMV. Natural infection of squash, pumpkin, cucumber, and watermelon by WMV as well as sesame was proved. The virus is inactivated at temperatures of 55 to , at dilution of , and in the aging of 10 to 14 days at about . Sesame, Chenopodium amaranticelor, pea, bean, as well as many plants of the Cucurbitaceae, are susceptible to the sesame-isolates of WMV. In negatively stained preparations, particles of the virus appear under the electron microscope as flexible filaments of about in length. Cylindrical inclusions and virus particles were found in the cytoplasm of mesophyll cells by ultra-thin sections of WMV infected tissues.

根瘤菌과 宿主植物의 相互作用에 관한 렉틴의 役割

張茂雄,全瓊姬,朴元學 한국응용곤충학회 1983 한국응용곤충학회지 Vol.22 No.4

강남콩 렉틴과 근류균(R. phaseoli)의 결합에 관한 광범위한 실험을 수행하여 다음과 같은 결과를 얻었다. 1. 강남콩종자의 자엽에서 가장 많은 렉틴은 성장에 따라 뿌리로 이동하였는데 발아후 약 5일경에, 뿌리의 길이가 인 것이 가장 렉틴의 함량이 많았다. 2. 근류균의 배양시기에 따가 강남콩렉틴과의 응집력을 측정 한 결과 배양초기 가 후기보다 응집력이 높았다. 3. 강낭콩종자에 존재하는 렉틴을 추출 정제하여서 6종 근류균과의 응집력을 측정한 결과 R. phaseoli와의 응집이 가장 강했고 이는 렉틴에 대한 특이적 결합, 즉 숙주특이성을 입증하는 것이었다. 4. 단당류에 의한 응집반응의 경쟁적 저해를 시도함으로써 강남콩렉틴의 근류균에 대한 결합부위가 mannose와 galactose를 포함한 oligosaccharides가는 것을 알수 있었으며 이들의 저해효과에 필요한 당의 최소농도는 6.25mM로서 측정되었다. 5. R. phaseoli와 강낭콩 렉틴을 강낭콩 뿌리에 인공감염시켜본 결과 렉틴의 경쟁적 저해가 확인되었으므로 뿌리혹형성에 렉틴의 가교가 관여한다는 가설을 입증할 수 있었다. 6. Immunodiffusion에 의한 R. phaseoli의 항원 결정기는 R. japonicum과 일부 관련성이 있을뿐 다른 Rhizobium spp.과는 전혀 다른 것으로 나타났다. Experiments were carried out to elucidate the specific interactions between host plant, Phaseolus vulgaris, and symbiotic bacteria, Rhizobium Phaseoli. Purified P. vulgaris lectins and six species of cultured Rhizobium were subjected to agglutination test. Lectins from bean and R. phaseoli showed relatively high agglutination activity indicating that host plant lectins recognize carbohydrate moieties on the compatible Rhizobium cell surface. The specific carbohydrate receptors for binding of the lectins on the cell surface of R. phaseoli were found as mannose and galactose. The minimum concentration of sugars for the inhibition was 6.25mM. The lectin content of cultured plant roots was measured after germination and was maximum in 5-day seedlings. The nodulation was competitively inhibited by lectins for the plants cultured with Rhizobium cells. By immunochemical studies, there was some relationship in antigenic determinants between R. phaseoli and R. japonicum but no relationships were observed with other Rhizobium species. The results suggest that the infection by rhizobia to the roots of leguminous plants may be caused by the specific interaction of lectins with rhizobia.

참깨의 모자이크 증상에서 분리한 수박${\cdot}$모자이크 바이러스에 관한 연구

장무웅,이창은,Chang M.U.,Lee C.U. 한국응용곤충학회 1980 한국식물보호학회지 Vol.19 No.4

참깨, 호박, 오이의 모자이크 증상주에서 전자현미경 및 즙액접종에 의하여 분리한 바이러스에 대하여 다음과 같은 결과를 얻었다. 1. 본 바이러스를 즙액접종 하였을 때 P. vutgaris, P. sativum, S. indicum, C. melo, C. maxima, C. pepo는 전신감염, C. amaranticolar는 국부감염 되었다. 2. 본 바이러스는 M. persicae로 쉽게 전반되었고, 종자전염은 하지 않았다. 9. 조즙액중의 물리성은 내열성이 $55\~60^{\circ}C$(10분), 희석성이 $10^{-3}\~10^{-4}$, 내보존성이 $10\~14$일$(22^{\circ}C)$이었다. 4. 본 바이러스의 형태는 길이 $750\~800nm$의 사상립자이고, 본 바이러스의 감염에 의해 세포질내에서 세포질봉입체가 만들어진다는 것이 확인되었다. 5. 본 바이러스에 감염된 S. indicum, C. amaranticolor의 병엽초박절편을 전자현미경으로 관찰하였던바, 각종세포의 세포질내에 사상립자가 병행배열 또는 산재해 있고, 또 사상립자와 함께 반드시 세포질내입체(pinwheel, boundles, laminated aggregates)가 확인되었다. 6. 이상의 결과를 종합해서 참깨 모자이크병의 병원바이러스를 Watermelon mosaic virus(WMV)로 동정하였다. This paper deals with the studies on the occurence of a new virus disease of sesame and the identification of the causal virus. The virus disease of sesame has been regarded as a widespread disease in the sesame-growing areas in the southern part of Korea. The disease was found to be caused by watermelon mosaic virus (WMV). During the years since 1978, stunting of sesame plants, with yellow mosaic, necrotic spot, and malformation, were collected from 17 different places. Virus isolates from 27 out of 32 samples were identified as WMV. Natural infection of squash, pumpkin, cucumber, and watermelon by WMV as well as sesame was proved. The virus is inactivated at temperatures of 55 to $60^{\circ}C$, at dilution of $10^{-3}\;to\;10^{-4}$, and in the aging of 10 to 14 days at about $20^{\circ}C$. Sesame, Chenopodium amaranticelor, pea, bean, as well as many plants of the Cucurbitaceae, are susceptible to the sesame-isolates of WMV. In negatively stained preparations, particles of the virus appear under the electron microscope as flexible filaments of about $750\~800nm$ in length. Cylindrical inclusions and virus particles were found in the cytoplasm of mesophyll cells by ultra-thin sections of WMV infected tissues.

은사시의 根頭癌腫에서 分離한 Agrobacterium tumefaciens에 關한 硏究

全桓惠,張茂雄 嶺南大學校 環境問題硏究所 1985 環境硏究 Vol.5 No.2

Agrobacterium tumefaciens (YPI, YPII, YPIII, YPIV, YPV) isolated from crown galls of Korean native populus tomentiglandulosa had characteristics as follows. Korean native strains appeared to be gram negative rod form having peritrichous flagella. Plasmids were isolated from YP strains by the alkaline method. The electrophoresis was performed with the plasmid band in 0.7% agarose gel. The experiment indicated that YPI had one plasmid, YPII had 5 plasmids, but YPIII, YPIV, YPV had no plasmid. The crude plasmids isolated from YPI and YPII strains were purified by CsCI-Et.Br density gradient centrifugation. The purified plasmids revealed open and supercoiled circular molecules under electron microscope. In electron microscopy of the ultra-thin sections of crown gall tissues, it was shown that bacterial cells were in the cytoplasm of crown gall cells and were surrounded by the membrane

심비디움 모자이크 바이러스의 핵산과 외피단백질에 관한 연구

배소현,장무웅 嶺南大學校 基礎科學硏究所 1994 基礎科學硏究 Vol.14 No.-

심비디움의 잎에 괴저 줄무늬 증상주에서 분리한 심비디움 모자이크 바이러스(CyMV-K)를 카틀레야와 팔레높시스에 증식시켜 순화 시료로 하였다. 순화된 CyMV-K는 길이 480-500nm, 폭 13nm의 약간 완곡형의 사상입자였다. 순화된 CyMV-K 시료의 자외선 흡광도를 조사한 결과, 260nm에서 최대, 248nm에서 최소값을 나타내는 전형적인 핵단백질의 자외선 흡수곡선을 보였다. Phenol/chloroform 추출에 의해 정제된 CyMV-K RNA의 분자량은 6.45Kb로 측정되었다. CyMV-K coat protein의 분자량 측정을 위해 SDS-PAGE법으로 전기영동한 결과, 분자량은 약 2.51 x 10⁴ 달톤으로 확인되었다. CyMV-K, CyMV-A, CyMV-J를 한천겔 이중확산법과 면역전자현미경법으로 혈청학적 관계를 조사한 결과, 혈청형이 거의 동일하였다. Cymbidium mosaic virus(CyMV-K) from Cymbidium sp, showing necrotic stripe symptom on the leaves was multiplied in Cattleya sp, and Phalaenopsis sp, and then purified. The purified CyMV-K was slightly flexible particles which have lengths in the range of 480-500 nm and widths of 13 nm. Ultraviolet absorption pattern of purified CyMV-K indicated the maximum at 260 nm and minimum at 248 nm, which was typical pattern of viruses. The molecular weight of CyMV-K RNA prepared by the extraction of phenol/chloroform was about 6.45 Kb. The molecular weight of CyMV-K coat protein estimated by SDS-PAGE method was about 2.51 x 10⁴daltons. CyMV-K, CyMV-A, CyMV-J was seorlogically identical by means of agar gel double diffusion test and immunosorbent electron microscopy.

관상용 구근류의 바이러스병

장무웅,김규원 한국화훼산업육성협회 1996 화훼연구 Vol.5 No.1

다알리아에서 分離한 3種의 바이러스

尹泰圭,張茂雄 嶺南大學校附設 基礎科學硏究所 1987 基礎科學硏究 Vol.7 No.-

대구와 진주에서 採集한 葉脈退綠, 葉脈?化. 綠色濃淡 모자이크, 退綠斑點, 委縮의 다알리아에서 3種의 바이러스가 分離되었다. 이들 바이러스는 電子顯微鏡, 寄主範圍, 血淸反應, 粗汁液中의 安定性, 진딧물의 傳搬性, 罹病葉組織內의 粒子 所在樣式에 의해 固定되었다. DMA: 汁液接種에 의해 백일홍에 全身感染(葉卷을 同伴한 委縮)이 확인되었다. 바이러스 粒子는 直經 50nm의 球形이다. 罹病葉切片의 電子顯微鏡의 관찰에서 葉肉細胞와 表皮細胞의 細胞質內에서 바이러스 粒子를 內包한 viroplasma가 확인되었다. BBWV: 이 바이러스는 汁液接種에 의해 8科 23種의 檢定植物에 感染이 확인되었다. 복숭아혹진딧물에 의해 쉽게 傳搬되었다. 耐熱性은 65-70℃, 耐稀釋性에서 ??, 耐保存性은 7-8日이였다. 바이러스 粒子는 直經 28nm의 小球形이다. 血淸試驗에서 이 바이러스는 BBWV의 抗血淸과 特異的인 反應帶를 形成했다. 罹病葉切片의 電子顯微鏡的 觀察에서는 各種細胞의 細胞質, 液胞內에서 바이러스 粒子의 集團 및 結晶像이 확인되었고, 특히 細胞質內에서는 vesicular body의 生成이 확인되었다. CMV: 이 바이러스는 汁液接種에 의해 Vignia sinensis의 接種葉에 傀疽斑點을 形成하였다. 바이러스 粒子는 直經 30nm의 小球形이다. 이 바이러스는 CMV의 抗血淸과 特異的인 反應帶를 形成했다. 罹病葉組織의 電子顯微鏡的 觀察에서는 葉肉細胞의 細胞質 및 液胞內에서 바이러스 粒子의 散在, 集團, 結晶配列像이 확인되었다. Samples of dahlia with chlorotic vein-banding, vein-clearing, yellow-green mosaic, chlortic mottling, malformation and stunting leaf symptoms were collected from dahlia-growing areas in Taegu and Chinju. Three viruses were distinguished under a electron microscope and their host range, serological reaction, stability in sap, type of aphid transmission, and relations with cells and tissues were examined. Dahlia mosaic virus(DMV) infection in zinnia y sap-inoculation produces dwarfing with downward leaf curling. In negatively stained preparations, particles of this virus appear under the electron microscope as spherical particle of 50nm in diameter. In ultrathin sections of DMV infected tissues, viroplasms were found in the cytoplasm of mesophyll cells and epidermal cells. Spherical particles, 40-45nm in diameter occurred within the dense granular matrix and electon-llucent areas of the viroplasm. Broad bean wilt virus(BBWV) was transmitted by sap-inoculation to 23 plant species in 8 families and by aphid, Myzus persicae. This virus was inactivated after 10 min at 65-70C, at dilution of ?? , and after 7-8 days at aout 20C. Electron microscopic examination of negatively stained preparation showed that the virus is spherical partile of 28nm in diameter. The virus reacted positively with BBWV-antiserum in agar gel deffusion test. In ultrathin sections of BBWV infected tissues, large aggregates or crystalline array of virus particles and vesicular body were found in the cytoplasm and vacuole of mesophyll cells. Cucumber mosaic virus(CMV) was transmitted by sap-inocultion. lnoculated leaves of Vignia sinensis produced necrotic local lesions. Electron microscopic examination of negatively stained preperation showed spherical particles of 30nm in diameter. The virus reacted positively with CMV-antiserum in agar gel diffusion test. In ultrathin sections of CMV infected tissues, large aggregates or crystalline array of virus of virus particles were found in the vacuole and a large number of virus particles were found in the cytoplasm of mesophyll cells.

부추와 마늘 바이러스의 分離 同定 및 理化學的 性質에 관한 硏究

노문철,박우원,장무웅 嶺南大學校 基礎科學硏究所 1991 基礎科學硏究 Vol.11 No.-

부추와 마늘은 바이러스 病에 대한 防止策없이 오랬동안 裁培되어온 韓國의 重要한 채소류 중의 하나이다. 따라서 이러한 바이러스 病을 防止하기 위해서는 부추와 마늘의 바이러스에 관한 基本的인 情報가 더욱 必要하다. 이러한 重要性때문에 本 硏究는 부추와 마늘의 바이러스를 分離 同定하여 그 特性을 調査하였다. 그 結果는 다음과 같다. 1. 부추와 마늘의 바이러스는 低速과 高速 遠心分離에 의해 純化되었고, 電子顯微鏡으로 觀察한 結果, 길이 650-700nm의 絲狀 粒子로 確認되었다. 2. 汁液 接種에 의해 感染된 명아주와 잠두의 잎은 局部病斑 症狀을 나타내었다. 3. 부추, 명아주, 잠두의 罹病葉을 초박절편하여 電子 顯微鏡으로 觀察한 결과, 바이러스 粒子는 集團 및 散在像으로 細胞質內에 存在하였다. 4. SDS-PAGE法으로 電氣泳動한 結果, 부추와 마늘의 바이러스의 coat protein의 分子量은 4.12×10⁴dalton으로 確認되었다. 5. 1.4% agarose gel 造成으로 水平電氣泳動한 結果, 부추와 마늘의 바이러스의 RNA의 分子量은 2.85×?? dalton으로 確認되었다. 6. GLV의 항혈청을 이용한 혈청학적 실험(면역전기영동법, 면역전자현미경법)에서 CCLV와 GLV가 동일한 반응을 나타냈었다. Chinese chive(Allium tuberosum) and garlic(Allium sativum) are an important vegetable crops for the korean people and have long been cultivated extensively in Korea. Because of this importance. studies were initiated to isolate and characterize the chinese chive latent virus(CCLV) and garlic latent virus(GLV). Result of these studies are summarized as follows: 1. CCMV and GLV were purified by alternate high and low speed centrifugation of chinese chive leaf or garlic leaf extracts. 2. Electron microscopic examination of CCLV and GLV revealed rod shaped. These paticles are measured about 650-700nm in length. 3. In manual inoculation, these viruses induced local lesion on Chenopodium album and broad bean. 4. In ultrathin section, Particles of CCLV and GLV were often found aggregate or scatered in the cytoplasm. 5. The molecular weight of the coat protein of CCLV and GLV were estimated by polyacrylamade gel electrophoresis in the presence of sodium dodecyl sulfate. Coat protein subunit of CCLV and GLV estimated about about 4.12×10⁴ daltons. 6. The moleculat weight of the nucleic acid of CCLV and GLV estimated about 2.85×?? daltons. 7. By the method of immunoelectrophoresis and immunosorbent electron microscope it has been proved that CCLV is identical to GLV.