Characterizations of Tobacco Mosaic Virus isolated from Chinese Foxglove(Rehmannia glutinosa Libosch)

지황(Rehmannia glutinosa Libosch)에서 분리한 Tobacco Mosaic Virus의 특성

  • Published : 2003.10.01

Abstract

This study was conducted to investigate the occurrence and characterization of tobacco mosaic virus(TMV) in Chinese foxglove isolated from the field of the Chonbuk province(Jinan, Jangsu, Jeongeup). TMV was detected in all three regions and confirmed positive reaction by ELISA test. In the host range test, Chenopodium amaranticola, Nicotiana glutinosa, N. tabacum cv. 'Bright yellow', N. tabacum cv. 'KY­57, Datura stramonium were locally infected with the virus. The virus produced mosaic symptom on inoculated leaves of N. tabacum cv. 'Samson'. However, Chenopodium quinoa, Glycine max, Raphanus sativus, Cucumis sativus, Cucurbita moschata, Brassica rape and Lycopersion esculentum did not show any symptoms. TMV particles were revealed as a stiff rod shape by transmission electron microscopic(TEM) and measured as 300 nm in length with 18 nm in diameter. Total RNA was extracted from showing symptom loaves infected with TMV and the reverse transcription­polymerase chain reaction (RT­PCR) obtained 531 bp DNA product of RNA with specific primer used. The capsid protein of TMV­RE showed higher amino acid sequence homology(97.7%) with TMV­To than with TMV­P(72.2%). The capsid protein of TMV­152 showed same amino acid sequence homology with TMV­F. The result of comparison of nucleotides sequence homology between TMV­RE strain and other TMV strain showed 94% homology with others except TMV­P(67.3%) and TMV ­ C(68.6%).

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.

Keywords

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