• Journal of Microbiology and Biotechnology
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• 제10권4호
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• pp.522-528
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• 2000

A total of about 300 fungal isolates from forest havitats were screened for inhibitors of tobacco mosaic virus (TMV) infection using its local lesion host, Nicotiana tabacum cv. Xanthi nc. Ine of the isolates, 14T, showed a strong activity against TMV infection, and was identified as an Apiocrea sp. based on its morphological characterstics. Rice was an optimum culture medium for its fermentation, and two antiviral compounds, KGT 141 and KGT 142, were resolved from the rice culture through column chromatography, TLC, and HPLC. By NMR and FAB-MS, the two compounds were identified as chrysospermins B (KGT 141) and D (KGT 142), both of which are peptaibols with 19-mer amino acids possessing an acetylated N-terminus and a hydroxy-amino acid (tryptophanol) at the C-terminus. Both compounds showed inhibitory activities against TMV infection, but chrysospermin D showed the stronger activity than chrysospermin B. The former of $100{\;}\mu\textrm{g}/ml$ and 54.7% at $10{\;}\mu\textrm{g}/ml$, respectively. Furthermore, the chrysospermins were highly cytotoxic toward cancer cell lines of PC-3 (prostrate) and K562 (leukemia), and inhibited growth of the Gram-positive bacteria tested, especially the plant pathogenic bacterium Corynebacterium lilium. To the best of our knowledge, this is the first report on the inhibition of plant virus infection by antimicrobial peptaibols.

• 식물병연구
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• 제8권2호
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• pp.101-106
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• 2002

시판중인 18품종의 토마토 종자에 대하여 TMV, ToMV, CMV의 오염률과 종자 전반률을 조사한 결과 TMV와 ToMV는 각각 9품종에서 오염이 확인되었으며, CMV는 3품종에서 바이러스 오염을 확인하였다. TMV는 3.8%로 가장 높은 오염률을 보였고, ToMV는 2.6%, CMV는 1.4%의 오염률을 보였다. 자연적인 종자 전반률을 조사한 결과, 공시한 9품종의 토마토종자에서 ToMV는 3품종에서 4.4%, CMV는 4품종에서 2.2%의 종자 전반률을 확인할 수 있었으며, TMV는 본실험에서는 종자 전반을 확인할 수 없었다. 1999년 10월부터 2000년 10월까지 충청북도 주요 토마토 재배지역에서 TMV, ToMV CMV의 지역별 바이러스 감염률과 병정별 감염률 및 생육시기에 따른 세 바이러스의 감염률을 조사한 결과, 각 지역의 토마토 포장에서는 잎말림, 괴저 및 왜화 등의 다양한 병징을 보였으며, 대부분의 포장에서 잎말림 증상이 관찰되었으며, 모자이크 증상과 누렁증상 등의 병징도 많이 관찰되었으며, 로꾸산마루 품종에서 다양한 병징이 관찰되었다. 지역별로는 옥산 지역의 로꾸산마루 포장과 가금지역의 꼬꼬 포장 및 노은지역의 로꾸산마루 포장에서 TMV와 ToMV 및 CMV의 감염률이 높았으며, ToMV의 감염률은 가금지역의 꼬꼬 포장에서 43.3%로 가장 높았으며, CMV의 감염률은 노은지역의 로꾸산마루 포장에서 40.0%로 다른 포장보다 높았다. 병징에 따른 바이러스의 감염률은 누렁증상을 보이는 병징에서 ToMV가 가장 높았고, 괴저 증상을 보이는 잎에서는 CMV가 가장 높았으며, 잎말림 과총생 및 얼룩무늬 등 대부분의 병정에서 ToMV의 감염을 확인하였고, TMV의 감염률은 낮았다. 생육단계별에 따른 바이러스 감염률을 조사한 결과 생육초기보다 생육중기와 후기가 바이러스의 오염률이 높았으며, 생육중기와 생육후기의 감염률은 차이가 없었다.

• 한국연초학회지
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• 제19권2호
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• pp.83-91
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• 1997

The vein-necrosis strain or potato virus Y (PVY-Vff) and black shank (Phytophlhora parasitica roar. nicotianae) causes severe damage on burley tobacco(Wicotiana tabacum L.) in Korea, A new burley tobacco resistance to PVY and black shank, KB 110, was developed by Korea Ginseng and Tobacco Research Institute. It was developed from the cross of Burley 21 with TC 591 in 1990, and was backrossed to Burley 21 in the following season. TC 591 has resistance to PVY and moderate resistance to race 0 of black shank, but it is susceptible to tobacco mosaic vim (TMV). KB 110 was evaluated for its resistance to PVY, TMV and black shank in the greenhouse and at fields for preliminary and performance trials. KB 110 which has secreting glandular trichomes was resistant to PVY-VN, TW and black shank. It had an erect growth habit and two more leaves per plant than that of Burley 21, and matures two to three days later. It yielded approximately 3 percent more cured leaf than the standard cultivar Burley 21, but other plant characteristics were very similar to those of Burley 21. It had acceptable standards for chemical and physical characteristics of lured leaf on regional farm test in 1995-1997. KB 110 produced average yields of good quality tobaccos and was appeared to be resistant to PVY inwhere occurrence of the virus are severe chronic at burley growing area.

• The Plant Pathology Journal
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• 제15권5호
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• pp.295-301
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• 1999

Many of plant defense responses are consequence of transcriptional activation of related genes. We have developed a modified differential screening procedure to isolate tobacco genes that are involved in the defense responses against TMV infection. A cDNA library was constructed from Nicotiana glutinosa leaves infected by TMV under temperature shift conditions. Each of plasmid DNA in the library was hybridized on a set of slot blots to a pool of cDNA probes prepared from either TMV-infected or mock-treated tobacco leaves. Among 900 plasmid DNAs, 81 clones exhibiting significantly enhanced or reduced level of hybridization to either probe were selected for nucleotide sequencing. The clones were listed into 61 genes considering redundancy between the sequences. The genes were identified to be defense-related genes including PR-genes and genes involved in primary or secondary metabolisms. This results supports the implication that plant defense process entails a major shift in total cellular metabolisms rather than activation of a limited number of defense-related genes. Expression patterns of a number of defense-related genes. Expression patterns of a number of selected genes were examined in northern blot analyses. It is notable that the clone 630 of unknown function exhibits expression pattern similar to those of previously known PR-genes. Experiments to elucidate the roles in defense mechanism of a couple of genes newly identified in this study are in progress.

• 한국연초학회:학술대회논문집
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• 한국연초학회 1997년도 담배과학 국제학술대회
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• pp.134-152
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• 1997

담배 모자이크 바이러스(TMV)와 감자 바이러스 Y(PVY) 등 식물바이러스병은 잎담배 생산에 심한 경제적 손실을 초래하고 있다. 바이러스병 방제를 위한 여러 가지 경종적 방법은 충분한-방제 효과를 얻지 못하는 경우가 많아 바이러스의 방제에는 우수한 저항성 품종의 사용이 가장 바람직한 것으로 알려져 있다. 그러나 기존의 육종 방법이 저항성 품종 개발에 항상 바람직한 것은 아닌데 그 이유는 저항성 유전자원이 없는 경우가 많고 또한 육종을 통해 유전자원의 열등한 특성이 도입될 수 있기 때문이다. 따라서 본 연구실에서는 TMV와 PVY의 여러 가지 형태의 외피단백질(CP) (비전사부분 포함 또는 제거, 비 전사부분 등) 및 복제유전자(Nlb) (3' 및 5' 결손 유무, 돌연변이)를 상용 담배 품종인 NC 82와 Burley 21에 형질전환시켜 바이러스 저항성 개발을 시도하였다. 각각의 유전자 cDNA를 1-2개의 35S promotor를 가진 식물발현벡타에 클로닝한 후 Agrobacteriupn (upnefaciens LBA 4404를 이용하여 식물의 잎조직에 도입시킨 후 kanamycin 함유 MS 배지에서 식물체를 재닥화하였다. 재분화 식물체는 TMV와 PVY에 대한 저항성 검정을 하였다. 그 결과 TMV에 저항성인 TMV CP 형질전환 식물체와 8 가지 Nlb 형질전환 식물체 계통 중에 6 계통의 저항성 형질전환 식물체를 획득하였다. 여기에서는 TMV와 PVY의 접종시험을 통하여 각각의 바이러스에 대한 형질전환 담배의 계통별 저항성 정도를 조사하고, 저항성 형질전환 식물체에서의 도입 유전자 확인하며, 세대별 저항성의 유전 및 안정성을 살펴보고자 한다. 또한 유망 저항성 형질전환 계통의 식물체 내에서의 바이러스 증식 및 이동과 관련된 저항성 기작, 여러 가지 PVY 계통에 대한 저항성 유무, 수량, 생육 특성 및 주요 화학 성분 함량 등을 발표하고자 한다.-glucose로 구성된 다당류 이었다. 아미노산은 Asp 및 Glu의 산성 아미노산과 Ala, Leu 등의 함량이 높게 나타났으며, 비알칼리 추출물에서 Ser과 Thr의 함량이 높게 나타났다. 다당류 T-AS는 평균 분자량이 2,000 kD와 12kD에서 주 peak를 나타냈으며, 수용성 분획의 평균 분자량은 12kD이고 비수용성 분획은 36~2,000 kD의 평균 분자량 분포를 갖는 것으로 나타났다. IR과 NMR 분석 결과 890 cm-1에서 흡수 peak를 나타내어 $\beta$-(1,3)0glucan과 $\beta$-(1,6)-glucan의 구조를 갖는 다당류로 확인 되었다. T-AS 분획은 C:H:O:N의 함량비가 38.9:5.7:49.6:1.84%이며, 이 물질의 융점은 163 $^{\circ}C$로 연한 갈색을 나타낸다. 분리된 GLG의 항암활성 기전 규명을 위해, in vivo 항암실험, 항보체 활성능, 항체 생성능, serum protein 분비능, 대식세포의 탐식능과 활성능 및 세포간 물질 분비 등의 상관관계를 조사하였다. 다당류 GLG 분획물들 가운데 항보체의 활성이 높았던 분획은 sarcome 180에 대한 항암 활성이 높게 나타났다. 다당류 T-AS의 보체 활성화 기작은 classical과 alternative complement pathway의 양 경로를 통해 활성화 되었다. T-AS 분획은 mouse내의 특정 혈청단백을 증가시켰으며, 항체 생성능의 증가가 관찰되어 effect T 세포의 활성화가 나타나고 있음을 알 수 있었다. T-AS는 생체내 투여시에 대식세포의 탐식능이 증진되었으며, 대식세포 기능 저해제에 의한 대식세포의 기능 저해 현상이 회복되었다. 이와 같은 결과들로부터, T-AS의 항암 활성은 활성화된 보체 성분 및 당 수용체들이 존재하는 대식세포의 개입을 시사한다.가능성과

• The Plant Pathology Journal
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• 제25권4호
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• pp.328-332
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• 2009

Diseases caused by Pepper yellow leaf curl virus infection is considered to be emerging plant diseases in Indonesia in the last five years. One key factor for disease management is the availability of accurate detection of the virus in plants. Polyclonal antibody for Pepper yellow leaf curl Indonesia virus-Bogor (PYLCIV-Bgr) was produced for detection of the virus using I-ELISA and DIBA methods. The antibody was able to detect PYLCIV-Bgr from infected plants up to dilution 1/16,384 and cross reaction was not observed with Cucumber mosaic virus (CMV), Tobacco mosaic virus (TMV), and Chilli veinal mottle virus (ChiVMV). Positive reaction was readily detected in membrane containing Begomovirus samples from Yogyakarta (Kaliurang and Kulonprogo) and West Java (Bogor and Segunung). Infection of PYLCIV-Bgr in chillipepper, tomato, and Ageratum conyzoides was also confirmed using polyclonal antibody for PYLCIV-Bgr in DIBA. Polyclonal antibody for PYLCIV-Bgr is suggested to be included in disease management approach due to its good detection level.

• 한국응용곤충학회지
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• 제15권3호
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• pp.137-145
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• 1976

우리나라에서의 바이러스 무감염란 생산에 필요한 기초자료를 얻기 위하여 Cymbidiun mosaic virus와 Tobacco mosaic virus를 중심으로 그 감염상 및 감염된 바이러스의 혈청학적 반응, 검정식물상의 반응, 물이적 성질, 형태등을 조사하여 다음과 같은 결과를 얻었다. 1. 란에 발생하는 바이러스병의 병징은 크게 1) 모자이크, 2) 괴저줄무늬 모자이크, 3) 괴원륜문, 4) 퇴록륜문 및 5) 괴달반점의 5군으로 분류되었다. 2. 수원근교의 난 재배농장에서 수집한 각종란의 바이러스 이병률을 조사한 결과 CyMV는 $45\%$의 감염율을 나타내었고, TMV는 한개체에서도 검출되지 않았다. 3. CyMV의 검정식물인 Chenopodium amaranticolor, Cassia occidentalis 및 Datura stramonium에 CyMV 병즙액을 접종한지 7-12일 후에 퇴록국부반점이 나타났다. 4. CyMV의 물리적 성질을 Chenopodium amaranticolor를 검정식물로 하여 조사한 결과, 내열성은$75-80^{\circ}C$ 내보존성은 8일, 내포석성은 $10^{-5}-10^{-6}$으로 나타났다. 5. CyMV의 혈정학적 한천내 확산반응에 사용된 0.1M 및 0.01M의 Phosphate, Imidazol 및 Tris 완충간에는 침강대 형성에 차이를 볼 수 없었다. 그러나 이들 완충액의 수소이온농도간에는 침강대 형성에 심한 차이를 나타내어, pH 7.0에서는 침강대가 뚜렷하였고 pH9.0에서는 미약하였다. 6. CyMV를 전자현미경으로 관찰한 결과 크기 460-580mu의 간상입자의 빈도가 제일 높았다.

• The Plant Pathology Journal
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• 제21권3호
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• pp.258-261
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• 2005

We conducted a survey on pepper virus diseases in 31 regions in Korea from November 2001 to December 2004. Using electron microscopy, test plant reaction, rapid immuno-filter paper assay (RIPA), reverse transcription-polymerase chain reaction (RT-PCR) and/or analysis of viral nucleotide sequences, we found a number of viruses from 1,056 samples that we collected. These included Cucumber mosaic virus (CMV), Pepper mottle virus (PepMoV), Pepper mild mottle virus (PMMoV), Broad bean wilt virus 2 (BBWV2), Tobacco mild green mosaic virus (TMGMV), and Tomato spotted wilt virus (TSWV). Of the samples analyzed, $343(32.5\%)$ were infected with CMV, $209(19.8\%)$ with PepMoV, $141(13.4\%)$ with PMMoV, $12(1.1\%)$ with BBWV2, $40(3.8\%)$ with TMGMV, $5(0.5\%)$ with TSWV, $153(14.5\%)$ with CMV and PepMoV, $54 (5.1\%)$ with CMV and PMMoV, $31(2.9\%)$ with PepMoV and PMMoV, $3(0.3\%)$ with CMV and BBWV2, $1(0.1\%)$ with CMV, PepMoV and BBWV2, $8(0.8\%)$ with CMV, PepMoV and PMMoV, and $30 (2.8\%)$ samples were infected with viruses which were not identified. CMV was the most predominant virus in all inspected fields and the number of the samples infected with PMMoV was relatively low as compared PepMoV infection level in pepper. TMGMV was only found in the southern part of Korea, while TSWV was isolated in Anyang and Yesan. However, we did not encounter in this survey the Alfalfa mosaic virus (AMV), Potato virus Y (PVY), Tobacco mosaic virus (TMV), and Pepper vein chlorosis virus (PVCV).

• The Plant Pathology Journal
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• 제26권4호
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• pp.328-339
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• 2010

A tobamovirus, Ribgrass mosaic virus (RMV), was identified newly from chinese cabbage (Brassica campestris L. pekinensis) in Korea. Virus disease incidence of RMV on chinese cabbage was 37.9% in alpine area on August in 1993. RMV induced the symptoms of necrotic ring spots, necrotic streak on midrib and malformation. RMV, Ca1 and Ca3 isolate, could infect 35 species out of 45 plants including Chenopodium amaranticolor. Physical properties of RMV Ca1 isolate were very stable as 10.8 over for dilution end point, $95^{\circ}C$ for temperature inactivation point and 18 weeks for longevity in vitro. RMV had the soil transmission rate of 75.0% for the chinese cabbages, 'Chunhawang' and 'Seoul' cultivars. The purified virions of RMV had the typical ultraviolet absorption spectrum of maximum at 260 nm and minimum at 247 nm. RMV of Ca1 isolate was related serologically with antisera of Tobacco mosaic virus (TMV)-Cym, TMV-O and Pepper mottle virus, but not related with antiserum of Odontoglossum ring spot virus. coat protein gene of RMV-Ca1, sized 473 nucleotides, encoded 158 amino acid residues. Nucleotide identity of RMV-Ca1 CP gene was 96.4% with RMV-Shanghai (GenBank accession No. of AF185272) from China and 96.0% with RMV-Impatiens (GenBank accession No. of AM040974) from Germany. Identity of amino acids between RMV-Ca1 and the two RMV isolates was 96.8%. Specific three primers were selected for rapid and easy genetic detection of RMV using Virion Captured (VC)/RT-PCR method.

• The Plant Pathology Journal
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• 제29권3호
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• pp.260-273
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• 2013

The incidence and distribution of Tobacco mosaic virus (TMV) and related tobamoviruses was determined using an enzyme-linked immunosorbent assay on 1,926 symptomatic horticultural crops and 107 asymptomatic weed samples collected from 78 highly infected fields in the major horticultural crop-producing areas in 17 provinces throughout Iran. The results were confirmed by host range studies and reverse transcription-polymerase chain reaction. The overall incidence of infection by these viruses in symptomatic plants was 11.3%. The coat protein (CP) gene sequences of a number of isolates were determined and disclosed to be a high identity (up to 100%) among the Iranian isolates. Phylogenetic analysis of all known TMV CP genes showed three clades on the basis of nucleotide sequences with all Iranian isolates distinctly clustered in clade II. Analysis using the complete CP amino acid sequence showed one clade with two subgroups, IA and IB, with Iranian isolates in both subgroups. The nucleotide diversity within each subgroup was very low, but higher between the two clades. No correlation was found between genetic distance and geographical origin or host species of isolation. Statistical analyses suggested a negative selection and demonstrated the occurrence of gene flow from the isolates in other clades to the Iranian population.