마늘 모자이크 바이러스에 관한 연구 -마늘 모자이크 바이러스의 분리, 검정식물상의 반응, 물리적성질, 순화, 혈청반응 및 전자현미경적관찰-

나용준,La Yong-Joon 한국응용곤충학회 1973 한국식물보호학회지 Vol.12 No.3

Garlic (Allium sativum L.) is an important vegetable crop for the Korean people and has long been cultivated extensively in Korea. More recently it has gained importance as a source of certain pharmaceuticals. This additional use has also contributed to the increasing demand for Korean garlic. Garlic has been propagated vegetatively for a long time without control measures against virus diseases. As a result it is presumed that most of the garlic varieties in Korea may have degenerated. The production of virus-free plants offers the most feasible way to control the virus diseases of garlic. However, little is known about garlic viruses both domestically and in foreign countries. More basic information regarding garlic viruses is needed before a sound approach to the control of these diseases can be developed. Currently garlic mosaic disease is most prevalent in plantings throughout Korea and is considered to be the most important disease of garlic in Korea. Because of this importance, studies were initiated to isolate and characterize the garlic mosaic virus. Symptom expression in test plants, physical properties, purification, serological reaction and morphological characteristics of the garlic mosaic virus were determined. Results of these studies are summarized as follows. 1. Surveys made throughout the important garlic growing areas in Korea during 1970-1972 revealed that most of the garlic plants were heavily infected with mosaic disease. 2. A strain of garlic mosaic virus was obtained from infected garlic leaves and transmitted mechanically to Chenopodium amaranticolor by single lesion isolation technique. 3. The symptom expression of this garlic mosaic virus isolate was examined on 26 species of test plants. Among these, Chenopodium amaranticolor, C. quince, C. album and C. koreanse expressed chlorotic local lesions on inoculated leaves 11-12 days after mechanical inoculation with infective sap. The remaining 22 species showed no symptoms and no virus was recovered from them whet back-inoculated to C. amaranticolor. 4. Among the four species of Chtnopodium mentioned above, C. amaranticolor and C. quinoa appear to be the most suitable local lesion test plants for garlic mosaic virus. 5. Cloves and top·sets originating from mosaic infected garlic plants were $100\%$ infected with the same virus. Consequently the garlic mosaic virus is successively transmitted through infected cloves and top-sets. 6. Garlic mosaic virus was mechanically transmitted to C, amaranticolor when inoculations were made with infective sap of cloves and top-sets. 7. Physical properties of the garlic mosaic virus as determined by inoculation onto C. amaranticolor were as follows. Thermal inactivation point: $65-70^{\circ}C$, Dilution end poiut: $10^-2-10^-3$, Aging in vitro: 2 days. 8. Electron microscopic examination of the garlic mosaic virus revealed long rod shaped particles measuring 1200-1250mu. 9. Garlic mosaic virus was purified from leaf materials of C. amaranticolor by using two cycles of differential centrifugation followed by Sephadex gel filtration. 10. Garlic mosaic virus was successfully detected from infected garlic cloves and top-sets by a serological microprecipitin test. 11 Serological tests of 150 garlic cloves and 30 top-sets collected randomly from seperated plants throughout five different garlic growing regions in Korea revealed $100\%$ infection with garlic mosaic virus. Accordingly it is concluded that most of the garlic cloves and top-sets now being used for propagation in Korea are carriers of the garlic mosaic virus. 12. Serological studies revealed that the garlic mosaic virus is not related with potato viruses X, Y, S and M. 13. Because of the difficulty in securing mosaic virus-free garlic plants, direct inoculation with isolated virus to the garlic plants was not accomplished. Results of the present study, however, indicate that the virus isolate used here is the causal v 현재 우리나라에서 재배되고 있는 마늘은 오랜 세월에 걸쳐 바이러스 감염에 무방비상태로 방치된채 영양번식을 계속해 왔기 때문에 대부분의 품종들이 퇴화되어 있을 것으로 생각된다. 따라서 마늘의 단위수량과 질의 향상을 기하기 위해선 바이러스 무감염 씨마늘의 육성, 보급에 의한 품종경신이 불가피할 것으로 보인다. 본 연구는 우리나라 마늘 재배지대 전역에서 가장 많이 발생하고 있는 모자이크병을 대상으로 바이러스의 분리, 검정식물상의 반응, 전염방법, 물리적성질, 순화방법, 혈청학적반응 및 형태등을 조사함과 동시, 마늘 모자이크 바이러스의 적절한 검정방법을 구명하여 앞으로 바이러스 무감염 씨마늘을 육성, 보급하는데 필요한 기초자료를 얻을 목적으로 실시했으며 다음과 같은 결과를 얻었다. 1. 1970-1972년까지 3년간 전투의 주요 마늘재배지대를 조사한 바 우리나라에서 재배되고 있는 마늘의 거의 대부분이 모자이크병에 걸려있음이 관찰되었다. 2. 마늘 모자이크 바이러스는 Chenopodium amaranticelor에 즙액접종 함으로써 단일계통을 분리할 수 있었다. 3. 26종의 검정식물을 공시하여 마늘 모자이크 바이러스를 즙액접종한 결과, 접종 11-12일 후에 Chenopodium amaranticolor, C. quince, C. album, C. koreanse 등 4종 식물의 접종엽상에 r국부병반 반응이 나타났다. 나머지 식물들에서는 병징이 나타나지 않았을 뿐 만 아니라 C. amaranticolor에 역접종했을 때도 바이러스는 회수되지 않았다. 4. 즙액접종에 의해 국부병반 반응이 나타날 상기 4종 Chenopodium속 식물중에서 C. amaranticolor 와 C. quinoa는 반응이 설민하고 정확하기 때문에 마늘 모자이크 바이러스의 검정식물로 적당하다고 생각된다. 5. 감염주에서 유내한 종구와 주아는 모두 모자이크 바이러스에 감염되어 있었고 이들 감염종구와 주아를 통하여 $100\%$ 전염되었다. 6. 마늘 모자이크 바이러스는 감염종구와 주아의 즙액에 의해서도 C. amaranticolor에 기계적 전염이 되었다. 7. C. amaranticolor 상에 계통분리된 마늘 모자이크 바이러스의 내열성은 $65^{\circ}-70^{\circ}C$, 희석한계는 $10^{-}2-10^{-3}$, 그리고 보존한계는 2 일이었다. 8. 마늘 모자이크 바이러스의 순화는 2회의 분획원심과 Sephadex gel filtration에 의해서 가능했다. 9. 전자현미경하에서 관찰한 마늘 모자이크 바이러스는 길이 1200-1225mu 폭 10-12mu의 사상이었다. 10. 혈청학적 미량침강 반응법에 의해 마늘잎에서뿐만 아니라 인편과 주아에서도 마늘 모자이크 바이러스의 검정이 가능했다. 11. 우리나라 5개 지방에서 수집한 마늘 종구 150개와 주아 30개에 대해 혈청학적방법으로 마늘 모자이크 바이러스의 감염률을 조사한 결과 $100\%$의 감염률을 보였다. 12. 마늘 모자이크 바이러스와 크기가 근사한 Potato Virus X. Potato virus Y, Potato virus S, Potato virus M 등과의 혈청학적 유연관계를 조사한 바, 마늘 모자이크 바이러스는 이들과 구별되는 다른 바이러스라고 생각된다. 13. 마늘의 모자이크 감염주에서 단일계통으로 분리하여 본 실험에 사용한 바이러스는 마늘의 바이러스 무감염주를 얻을 수가 없기 때문에 직접 마늘잎에 접종해서 모자이크톤의 병원이라는 것을 확인할 수 없었지만, 검정식물상의 반응, 혈청학적반응, 전자현미경적 관찰등의 간접적인 조사 결과로 미루어 미인록의 마늘모자이크 바이러스라고 생각된다.

면역이중확산법에 의한 콩 종자의 모자이크 바이러스(SMV) 감염상 조사

나용준,박원철,오정행,La Yong-Joon,Bak Won-Chull,Oh Jeung-Haing 한국응용곤충학회 1983 한국식물보호학회지 Vol.22 No.1

우리 나라 콩 종자의 모자이크 바이러스(Soybean mosaic virus; SMV) 감염상을 조사하기 위하여 국내에서 수집한 8개 품종을 공시해서 Lima와 Purcifull의 면역이중확산법으로 SMV 검정을 한 결과는 다음과 같다. 1. 공시한 7개 품종 중 6개 품종에서 SMV가 검출됨으로서, 우리 나라의 콩 품종에서 SMV의 종자전염성이 혈청학적 방법으로 확인되었다. SMV가 검출된 종자의 종자감염성율은 최저 $2.1\%$에서 최고 $12.5\%$를 나타냈으며, 전체적으로는 총검정립수 336립 중 18립에서 SMV가 검출됨으로서 약 $5.4\%$의 감염율을 보였다. 2. 이병주에서 채집한 갈반립과 무갈반립수의 SMV 감염율은 북해 1호 품종의 경우 각각 $33.3\%$와 $29.2\%$ Clark 품종의 경우 각각 $4.2\%$와 $0.0\%$, Woodworth 품종의 경우는 갈반립과 무갈반립 모두 $4.2\%$의 감염율을 나타냄으로써, 공시한 콩 품종의 갈반립과 무갈반립간엔 SMV 감염율에서 뚜렷한 차이를 발견할 수 없었다. 3. SMV에 감염된 광교품종 중 괴저병징을 나타내는 개체에서 채종한 종자에서는 전혀 SMV가 검출되지 않아 광교품종에서는 SMV-N가 종자전염되지 않는 것으로 보였다. Soybean mosaic virus (SMV) infection in seeds wits investigated by an immunochemical method Seven soybean cultivars or lines Bughae No.1, KAS 662-7, Chungbugbaeg, Gwanggyo Clark, Bongeui, and Gangrim were tested using hypocotyls of germinated seeds and presence of SMV was detected in six soybean cultivars but Gangrim. The level of SMV infection in the assayed cultivars varied from 2.1 to $12.5\%$. It seemed that seed coat mottling had no correlation with seeds. SMV infection of the seeds since virus has not always been detected from the mot tled seeds SMV has not been detected in the seeds of variety Gwanggyo which showed necrotic symptoms.

한국에서의 포플러 모자이크 바이러스 검출

나용준,이창근,여운홍,황재우,심상영 ( Yong Joon La,Chang Keun Yi,Woon Hong Yeo,Jae Woo Hwang,Sang Yung Shim ) 한국산림과학회 1980 한국산림과학회지 Vol.49 No.1

Several plants of Populus deltoides `R-89` exhibiting symptoms of poplar mosaic described earlier by several researchers (1,2,3) were observed in the poplar nursery of the Institute of Forest Genetics at Suweon, Korea during September, 1980. The symptoms produced a mosaic or discolored yellow spottings in mature leaves with veinal and petiolar necrosis and leaf curling (Fig. 1 and 2). Leaves with mosaic symptoms were collected and leaf-dip preparations from chlorotic area were examined for the presence of virus particles with an electron microscope. Electron micrographs of the negatively stained leaf-dip preparations revealed numerous elongated virus particles ranging 200-1300nm in length (Fig. 3). Of 156 panicles measured randomly, 68 particles (43.6%) were in the range of 660-670nm in length (Fig. 4). The values obtained fell well within the range of those described for poplar mosaic virus (PMV) by other researchers (1. 2, 3). No viruslike particles were observed from poplar leaves without mosaic symptoms. Further studies on the properties of the PMV isolate and control of the disease are underway.

감자 바이러스의 혈청학적 동정에 관한 연구

나용준,La Yong-Joon 한국응용곤충학회 1974 한국식물보호학회지 Vol.13 No.1

우리나라 씨감자에 발생하고 있는 잠복성바이러스의 종류와 이들의 감염상태를 조사하기 위하여 감자 원원종포, 원종포 그리고 원예시험장 등에서 수집한 외관상 무병 건전하게 보이는 감자와 외관상 바이러스병징이 뚜렷한 감자를 대상으로 혈청학적방법에 의해 바이러스 검정을 실시했다. 검정대상 바이러스는 Potato Virus X(PVX), Potato Virus S(PVS), Potato Virus M(PVM), 그리고 Potato Virus Y (PVY)의 4종이며, 바이러스의 검정은 van Slogteren의 미량침강법에 따라 실시했다. 결과는 다음과 같다. 1. 씨감자 생산포장에서 수집한 감자잎에서 PVX, PVS, PVM, 그리고 PVY가 검출되었는데 우리나라 감자에서 PVM이 검출되었다는 실험적 보고는 이것이 처음이다. 2. 외관상 건전하게 보이는 씨감자의 거의 $100\%$ PVX와 PVS에 약 $10\%$가 PVX+PVS+PVM에, 약 $4.5\%$가 PVX+PVS+PVY에, 그리고 $1\%$ 정도가 PVX+PVS+PVM+PVY에 각각 혼합감염되어 있었다. 3. PVX와 PVS만이 검출된 식물은 대부분 병징이 음폐되어 있었으나 PVX, PVS와 함께 PVY가 검출된 식물은 대부분 뚜렷한 병징을 나타내고 있었다. 4. 혈청학적 미량탐강법은 육안적인 관찰만으로는 검정이 어려운 PVX, PVS, PVM 및 mild strain의 PVY를 씨감자 생산지에서 대규모의 식물은 대상으로 신속정확히 검정하는데 애우 편리한 방법으로 생각된다. A total of 230 apparently healthy looking potato stocks and 80 potato stocks with symptoms of virus infection were collected from various seed potato farms in Korea and the incidence of potato virus X (PVX), potato virus S (PVS), potato virus M (PVM) and potato virus Y (PVY) was determined by serological microprecipitin tests. Results obtained are as follows. 1. Serological microprecipitin test retreated the presence of PVX, PVS, PVM and PVY in a number of potato stocks grown for the production of seed potatoes in Korea. 2. The occurrence of potato virus M is reported here for the first time in Korea with experimental evidence. 3. Practically $100\%$ (290 stocks, of the apparently healthy looking potato stocks were demonstrated to be infected with both PVX and PVS. The infection percentages of potato stocks with combination of PVX, PVS, PVM and PVY were as follows. PVX+PVS+PVM:$10.3\%$, PVX+PVS+PVY:$4.5\%$, PVX+PVS+PVM+PVY:$1.03\%$ 4. Irish Cobbler and Shimabara, which are the two major potato varieties in Korea, appear to be symptomless carriers of PVX and PVS. However, when these varieties were infected additionally with PVY, usually severe symptoms resulted. 5. Serological microprecipitin technique appears to be highly suitable for early, quick and reliable diagnosis of PVX, PVS PVM and PVY. It is particularly suited for large scale testing of seed potato stocks for the presence of viruses mentioned above.

옥시테트라사이클린에 의한 오동나무 · 빗자루병 방제 (防除)

나용준,신현동 ( Yong Joon La,Hyeon Dong Shin ) 한국산림과학회 1980 한국산림과학회지 Vol.49 No.1

The witches` broom disease of Paulownia tomentosa (Thunb.) Steud with which mycoplasmalike organisms are associated is widespread throughout Korea and poses serious threat to the cultivation of paulownia. Attempt was made to investigate the feasibility of field control of the disease with oxytetracycline hydrochloride (OTC). A total of 84 paulownia trees (6 year-old, DBH: 10-15㎝) exhibiting severe symptoms of witches` broom were selected and treated during March to September. Solution of 1-10g of OTC dissolved in 0.1­2.0 of water was transfused into infected trees with gravity flow method from a dark-brown colored plastic reservoir (11 volume) through plastic tubes (1.2m long) connected to 2-4 holes (5 ㎜ in diameter and 4-5㎝ in depth) bored in the basal part of the tree trunks (Fig. 1 and 2). Of 60 diseased paulownia trees injected with 2g of OTC in 0.1-2.01 of water during May to September, 1979, 58 trees resulted in complete remission of symptom development and resumption of healthy new growth at least up to September, 1980 when the last observation of the effect of OTC treatment for this experiment was made. The rest of two trees were dead probably due to too severe infection. Of 24 paulownia trees treated in March and April, 1979 complete remission of symptom development was obtained with 8 trees, and nine trees were partially prevented from symptom development in the following season. The remaining 7 trees were dead due to failure in uptake of OTC and partly because the trees were in too far advanced stage of infection. Application of highly concentrated solution of 2g of OTC dissolved in 0.1-0.21 of water per tree was just as effective as the 2g/1-2l treatment. Injection of 2g/1-2l required 3-4 days while treatment of 2g/0.1-0.2l reduced the time for injecting one tree down to less than 24 hrs. The result of this experiment demonstrates that basal trunk injection of 2g OTC/0.1-0.2l/tree is feasible for field control of paulownia witches` broom, provided that tree injection is performed in actively growing season (May-September) and at the initial stage of disease development.

Current Status of Researches on Mycoplasmal Diseases of Plants in Korea

나용준,La Yong Joon The Korean Society of Plant Pathology 1987 Plant Pathology Journal Vol.3 No.1

빗자루병에 감염된 대추나무 조직내의 마이코플라스마 분포

나용준,이덕재 ( Yong Joon La,Deok Jae Lee ) 한국산림과학회 1984 한국산림과학회지 Vol.67 No.1

Distribution of mycoplasma-like organisms (MLO) in the phloem tissue of witches`-broom infected jujube trees was investigated by fluorescence microscopy applying new fluorochrome DAPI (4`-6-diamidino-2-phenylindole, 2HCl). MLO were detected from the phloem of leaf and stem sections of diseased branches exhibiting typical witches`-broom symptom but not detected from those of symptomless branches of an infected tree. MLO were also present in the healthy looking leaves frequently found in the diseased shoots. Fluorescence microscopy revealed the presence of MLO in the phloem of root sections of infected trees.

Oxytetracycline 수한주입에 의한 대추나무 비짜루병 방제

나용준,문동식,La Yong-Joon,Brown William M.,Moon Dong-Sik 한국응용곤충학회 1976 한국식물보호학회지 Vol.15 No.3

oxytetracycline HCI의 대추나무 비짜루병 방제효과를 구명할 목적으로 병든 대추나무(15-20년생, 흉고직경 20cm 내외)에 oxytetracycline HCI의 1,000 p.p.m. 수용액을 1회에 500ml씩 시기별로 각각 1회(4월), 2회(4월, 7월), 30(4월, 6월, 8월) 그리고 대추 수확후 1회구(10월)로 나누어 수간에 주입하여 다음과 같은 결과를 얻었다. 1. 수간주입당년에 있어서는 시기별 주입회수에 관계없이 1회 이상의 수간주입으로 비짜루병징이 완전히 억제되고, 정상엽이 자라나면서 개화결실하였다. 2. 2회이상 주입한 나무와 10월에 1회주입한 나무에서는 이듬해 가을까지도 병징억제효과가 지속되었으나 4월에 1회 주입한 나무에서는 이듬해에 부분적으로 경미한 병징이 나타나기 시작했다. 3. oxytetracycline HCI에 의한 병징억제효과는 수간주입부위의 윗쪽에서만 나타났으며, 그 이하부위와 약액이 주입되지 않은 측지에서는 병징이 억제되지 않았다. 4. oxytetracycline HCI의 5.000 p.p.m. 수용액을 주당 30ml씩 주입하였을 때는 전혀 병징이 억제되지 않았다. A Witches'-broom disease of jujube (Zizyphus jujuba) with which mycoplasma-like bodies are associated is wide spread in South Korea. Jujube trees with witches'-broom symptoms were selected from 15-20 year old planting and treated with solution of oxytetracycline hydrochloride (Terramycin). Treatments were 1) one injection on April; 2) one postharvest injection on October; 3) two injections (April and July); 4) three injections (April, June and August) and 5) an untreated control. Each injection consisted of 500ml of 1,000 p.p.m. solution of oxytetracycline HCl transfused into affected trees from plastic reservoir through plastic tubes connected to 3 small holes drilled in the basal part of the tree trunks. Complete remission of witches'-broom symptoms was accomplished within one growing season by one spring (April) injection. One postharvest (Oct.) injection also prevented the symptoms in the following growing season. Two and three injection treatments prevented the symptoms for at least two growing seasons and restored previously severely diseased trees to normal or near normal conditions. Remission of symptoms was found only above the injection site while current season witches'-brooms developed from areas below the injection site and from untreated main scaffolds. The results of this experiment demonstrates that transfusion treatment with oxytetracyclin HCl is feasible for the practical control of witches'-broom of jujube.

빗자루병에 감염된 대추나무 조직내 단백질의 전기영동 (電氣泳動)

나용준,박원철,박원목,이용세 ( Yong Joon La,Weon Chul Park,Weon Mook Park,Yong Se Lee ) 한국산림과학회 1984 한국산림과학회지 Vol.66 No.1

Protein profiles of healthy and witches` broom (mycoplasma) infected jujube plants (Zizyphus jujuba) were investigated by 2-30% linear gradient polyacrylamide gel electrophoresis. Distinct differences in band patterns between healthy and infected samples were observed. Gels from samples of healthy leaves showed a characteristic protein band in the 50kd-range, which was not detected in infected leaves. Band with 25kd was more distinct in the infected leaves, whereas band with 198kd was more apparent in the healthy leaves. Healthy-looking leaves in the infected samples demonstrated the characteristic band of 50kd with less intensity showing intermediate pattern between healthy leaves and infected leaves. In contrast with leaf samples, gels from infected stem samples showed a characteristic band in the 335kd-range which was absent in healthy stem samples.

Latex 응집반응법에 의한 고추의 담배 모자이크 바이러스 검출

나용준(La Yong-Joon),윤수경(Yoon Soo-Kyung),김병환(Kim Byung-Hwan) 한국원예학회 1994 한국원예학회 학술발표요지 Vol.12 No.1