• 식물조직배양학회지
• /
• 제25권4호
• /
• pp.245-250
• /
• 1998

Tobacco mosaic virus(TMV) 외피단백질 유전자를 연초(Nicotiana tabacm cv. NC82)에 형질전환하고 형질전환 식물체 후세대에서 TMV 저항성인 연초를 선발하여, 선발된 TMV 저항성 제5세대 형질전환 식물체의 도입된 유전자발현 및 고온에서의 특성 등을 조사하였다. TMV 저항성 식물체의 염색체 DNA에 TMV 외피 단백질 유전자가 안정되게 존재하고 있음을 genomic PCR을 수행하여 확인하였다. 또한 형질전환 식물체내에서 TMV 외피 단백질 발현은 Immunoblot hybridization 방법으로 확인하였다. TMV 저항성 형질전환 연초식물체에서 발현된 단백질의 양은 매우 적었으며 특히 본엽에는 병징이 나타나지 않았으나 수확기 마지막 액아에 TMV의 반점이 나타난 병징발현 지연형의 형질전환 식물체의 경우에도 발현된 단백질의 양은 정상 NC 82에 TMV가 감염되었을 때와 비교하여 현저히 적었다. TMV 저항성 형질전환 식물체 내에서 발현되는 TMV 외피단백질의 양은 총 단백질에 대비하여 0.01% 이하이였다. TMV 병징 발현 지연형인 형질전환 식물체에 TMV를 인공접종한 후 고온처리상태에서 외피 단백질 유전자의 전사 및 발현을 RT-PCR과 Immune blot hybridization 통하여 확인하였으며, 이때 TMV의 증식도 억제되었으므로 개량멀칭시 나타나는 고온조건하에서도 저항성이 안정적으로 발현될 수 있음을 알 수 있었다.

• 한국식물병리학회지
• /
• 제11권1호
• /
• pp.87-90
• /
• 1995

Complementary DNA of the movement protein (MP) gene of tobacco mosaic virus Korean pepper strain (TMV-KP) was synthesized from purified TMV-KP RNA by using the reverse transcription and polymerase chain reaction (PCR) system. The synthesized double stranded cDNA was cloned into the plasmid pUC9 and transformed into Escherichia coli JM110. The movement protein gene of TMV-KP of the selected clones was subjected to sequence analysis by Sanger's dideoxy chain termination method. The complete sequence of viral MP gene from TMV-KP strain was 807 nucleotides long. The nucleotide of MP gene from TMV-KP has thirteen and two nucleotide differences from TMV vulgarae (TMV-OM) and Korean (TMV-K) strains, respectively. Thus, the nucleotide sequence of TMV-KP MP gene showed higher homology of 99% with that of TMV-K MP gene.

• 한국식물병리학회지
• /
• 제14권4호
• /
• pp.319-324
• /
• 1998

Induction of systemic acquired resistance (SAR) against phytophthora blight and pathogenesis-related (PR) protein accumulation by TMV infection in pepper plant (Capsicum annuum cv. Nockwang) were examined to understand the mechanism of the systemic acquired resistance in pepper plant. The zoospore suspension of Phytophthora capsici was inoculated on stem of pepper plant in which TMV-pepper strain had been inoculated on fully expanded upper leaves, and thephytopha blight incidence was examined. Both disease severity and lesion length of phytophthora blight were much smaller in TMV pre-inoculated pepper plant than in uninoculated control plants. The phytophthora blight incidence was decreased about 50% in the TMV pre-inoculated pepper, compared to the uninoculated control plant at 10 days after P. capsici inoculation. Accumulation of PR1 and PR5 proteins in intercellular fluid of TMV-inoculated and uninoculated upper leaves were monitored by immuno-blot with tobacco P1b and PR5a, antibody during induction of SAR. PR1 and PR5 were detected from 24 hours after TMV inoculation in both TMV-inoculated and uninouclated upper leaves, and increased rapidly in TMV-inoculation in uninoculated upper leaves were defoliated. PR5 could be detected upto 20 days after TMV inoculation in uninoculated upper leaves. These results suggest that TMV infection induces SAR against phytophthora blight in pepper plant, and that PR proteins are accumulated very rapidly during induction of SAR and maintained for quite long time in pepper plant.

• 한국식물병리학회지
• /
• 제14권5호
• /
• pp.371-375
• /
• 1998

Tobacco (Nicotiana tabacum) cultivars including NC 82 and KF 114, and Datura stramonium, Physalis floridana, Gomphrena globosa, and Chenopodium spp. were added to the previous host plants tested for the further examination on the biological characteristics of tobacco mosaic virus (TMV) strains isolated from tobacco (TMV-Common), tomato (TMV-Tomato), and pepper (TMV-Pepper), In TMV-Tomato and TMV-Pepper, different symptoms were noted in P. floridana (no symptom development), and NC 82 (local lesion production on the inoculated leaves) from TMV-Common with which systemic mosaic symptoms were developed. Local lesions were developed in KF 114, D. stramonium, G. globosa, and Chenopodium spp by TMV-Common and TVM-Tomato, while no symptom was observed in KF114 and G. globosa. Also the number and size of local lesions were smaller in KF 114 than Xanthi-nc tobacco (local lesion host) infected with TMV-Tomato. Systemic necrosis was induced in Xanthi-nc and KF 114 when infected with TMV-C at high temperature, but not with the other strains.

• 한국자원식물학회지
• /
• 제16권3호
• /
• pp.230-237
• /
• 2003

전라북도의 진안, 장수, 정읍 등의 3개 지역에서 재배하고 있는 지황에 바이러스 병징이 관찰되어 ELISA 방법으로 TMV의 감염 여부를 확인한 결과 3개 지역 모두에서 ELISA 양성 반응을 나타내었다. N. glutinosa를 이용, 순수분리 및 증식한 이병주의 바이러스 입자를 관찰한 결과, 폭 18nm, 길이 300nm의 막대 모양의 바이러스가 관찰되었다. 또한 epoxy수지를 통해 이병조직의 세포질내에서 많은 바이러스 입자가 군집 또는 산재되어 있음이 관찰되었다. TMV의 병징은 즙액 접종 4주 후부터 담배 잎의 본엽에 부분적으로 반점이 보이다가 4­6주 후에는 황화 현상과 괴사 현상이 나타났다. 이병주로부터 total RNA를 분리하여 RT­PCR을 수행한 결과 531bp DNA 증폭 산물을 얻었으며 genetyx win program을 이용하여 외피단백질의 염기수준에서 비교 분석한 결과, TMV­T, V, To 세 계통에서 각각 94.83％로 가장 높게 나타났고, TMV­P계통에서 67.35％로 가장 낮게 나타났다. 아미노산 수준에서는 TMV­T, V, To 세 계통에서 각각 97.65％로 가장 높게 나타났고, TMV­P계통에서 72.22％로 가장 낮게 나타났다. 기타 계통들간에는 TMV­152, F 계통에서 94.05％, TMV­C 계통에서 68.63％ 유사도를 나타냈다.

• 한국연초학회지
• /
• 제24권1호
• /
• pp.7-12
• /
• 2002

During the screening or antiviral substances having inhibitory effect on tobacco mosaic virus(TMV) infection to tobacco plants, we found that a bacterial isolate, KTB61, which was identified as a Pseudomonas sp., strongly inhibited the formation of TMV local lesions. When the culture filtrate from KTB61 was applied on the upper surface of leaves of N. tabaccum Xanthi-nc tobacco at the same time of or 24 hours before TMV inoculation, almost complete inhibition was achieved. Incidence of systemic TMV infection to the susceptible tobacco cultivar, NC82, was reduced by 95% when TMV was inoculated onto the upper surface of leaves 24 hours after spraying the culture filtrate. Also 75∼80% of inhibitory effect was obtained by the inoculation of TMV onto the under surface of the leaves treated with culture filtrate 24 hours beforehand. In field trials, the TMV infection was reduced by 96.5% when the tobacco seedlings, N. tabaccum cv. NC82, were soaked with culture filtrate before transplanting.

• 한국연초학회지
• /
• 제2권1호
• /
• pp.53-60
• /
• 1980

우리나라 煙草耕作地의 담배모자이크 바이러스(TMV)病의 時期別 感染率 變化를 調査하고 傳染源 究明을 爲하여 越冬後 담배와 고추뿌리, 雜草類, 담배 果皮에서 生物學的 및 血淸反應에 따라TMV를 檢定하였던바 그 結果를 要約하면 다음과 같다. 1. 本國에서 TMV 初期感染은 改良말칭栽培의 경우 移植 및 一般말칭으로의 轉換時, 一般말칭栽培의 의 경우는 移植 및 1次 腋弟除去時가 가장 重要한 時期로 判斷된다. 2. 越冬後 土壤中의 담배 및 고추뿌리에서 活性 TMV가 檢出되었으며, 부기된 뿌리일수록 그 濃度가 낮았다. 3. 22科 38種의 雜草를 檢定한 結果 TMV에 自然感染되어 있는 것은 까마중과 꽈리 2種이었다. 4. 담배의 果皮에서 TMV가 檢出되었으며 어린 葯에서는 感染이 認定되지 않았다.

• The Plant Pathology Journal
• /
• 제15권4호
• /
• pp.247-250
• /
• 1999

Wasabies showing mosaic symptoms were collected and extracted for virus purification. Tobacco mosaic virus (TMV) was identified as causal agent by electron microscopy and nucleic acid and coat protein analyses. TMV strains were determined by enzyme-linked immunosorbent assay (ELISA). TMV was identified as W and C strain in wasabi. The results of host reaction indicated that this virus induced local lesions on Nicotiana tabacum cv. Bright Yellow and N. glutinosa, leaf spots on Chenopodium amaranticolor and mosaic symptoms on wasabi. Rot shape virus particles were observed and was about 300 nm in length. About 6.5 kb single RNA molecule was observed from extracted viral RNA sample and 26 KDa coat protein was detected in denatured acrylamide gel. Infection ratio of TMV was 8% for the first cultivation year, but was 22% for the second year when TMV-W antiserum was used. The results of this experiment showed that infection ratios of both TMV-W and TMV-C strains were higher compared to that of TMV-P strain.

• 한국연초학회지
• /
• 제21권1호
• /
• pp.70-76
• /
• 1999

KF 116 was TMV resistant tobacco plant and KB 301 was PVY resistant plant transformed with TMV CP gene and PVY CP gene, respectively. These resistant plants were cross-fertilized and the 4 lines of the TMV-PVY resistant plants were selected from F1 hybrid plants. The rate of PVY-resistant plant in these hybrids was 100 percent and that of TMV-resistant plants including delay type was 90-98 percent at 4 weeks after virus inoculation. It was confirmed that the TMV and PVY CP genes were integrated into the genome of hybrid plants by genomic PCR, and Southern blot hybridization. The genome of F1 hybrid plants had one copy and 4 copies of PVY-CP gene and TMV-CP gene, respectively, and CaMV 35S promoters were not methylated, regardless of the difference symptom development to TMV.

• 식물병연구
• /
• 제9권2호
• /
• pp.89-93
• /
• 2003

남해지역의 원원종 종서 생산 포장에서 '추백' 품종에 나타난 엽맥투명 및 매우 약한 모자이크 증상을 나타내는 감자 잎에서 담배 모자이크 바이러스(TMV)를 분리하였다. 이 바이러스((TMV-St))는 생물학적, 혈청학적 유연관계 및 외피단백질의 염기서열 등을 통해 기존에 보고된 다른 tobamovires와 비교하였다. TMV-St는 5개의 지표식물 반응에서 토마토, 고추, 가지 등과 같은 가지과 작물에 경제적 피해를 주고 있는 TMV-U1, Pepper mild mottle virus(PMMoV) 및 Tomato mosaic virus(ToMV)와는 다른 기주 반응을 보였다. 특히 즙액접종에 의한 기주의 반응은 C.murale 접종엽과 상엽 모두에서 퇴록반점을 보였으며, C. murale, G. globosa, N.rustica 그리고 N. tabacum ce. Samsun nn 등 4가지 지표식물로 이들 바이러스 계통을 구분할 수 있었다. 혈청학적 검정에서 TMV-St는 TMV-U1, PMMoV 그리고 ToMV와의 반응에서 도두 뚜렷한 침강선을 형성하였다. TMV-St의 외피단백질은 477개의 염기서열로 되어 있으며, 이는 TMV-U1의 염기서열과 매우 유사하였다.