Molecular cloning of recalcitrant plant viruses genes by a modified RAPD method using RF-dsRNA
| Project/Area Number |
12660041
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
植物保護
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| Research Institution | Utsunomiya University |
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Principal Investigator |
NATSUAKI Tomohide Utsunomiya Univ., Fac.Agri., Professor, 農学部, 教授 (10134264)
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2002: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2001: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2000: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | recalcitrant plant viruses / replicative form dsRNA / PCR / simple detection method / molecular cloning / 難純化RNAウイルス / キュウリ黄化ウイルス / tomato infectious chlorosis virus / Cryptovirus / カンキツトリステザウイルス |
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| Research Abstract |
Molecular cloning and sequencing of plant virus genes has been carried out during the past decade. One objective of molecular analysis of plant viruses has been the improvement of the methods to detect and diagnose pathogenic viruses. As templates for Cdna synthesis, nucleic acids are usually extracted from purified virus preparations in relatively pure form and in rather large amounts. Theses strategies rely on the purification of virus particles from infected plants. However, there are many recalcitrant viruses that can not be purified by current methods and, therefore, the standard nucleic acid templates are not accessible for their cloning. It is the viruses for which there are no available antisera that alternate methods of detection and diagnosis are needed. For several of these viruses, the application of dsRNA extraction techniques from herbaceous or woody host plants has permitted the detection of viral replicative nucleic acids (RF-dsRNA). The main objective of this research was the production of cDNA clones generated from RF-dsRNA extracted from virus-infected plant tissues. In this research the simple one tube RT-PCR method to detect these viruses was established. Furthermore, full nucleotide sequences of mild and severe isolates of citrus tristeza vireus and Cucumber yellows virus were determined. Tomato infectious chlorosis virus, a member of Criniviruses, was newly recognized in Japan.
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Report
(4 results)
Research Products
(11 results)
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[Publications] Suastika,G., Natsuaki,T., Terui,H., Kano,T., Ieki,H. and Okuda,S.: "Nucleotide sequence of citrus tristeza virus seedlding yellows isolate"Journal of General Plant Pathology. 67-1. 73-77 (2001)
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Publications] Suastika, G., Natsuaki, T., Terui, H., Kano, T., Ieki, H., Okuda, S.: "Nucleotide sequence of citrus tristeza virus seedling yellows isolate"Journal of General Plant Pathology. 67・1. 73-77 (2001)
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[Publications] Suastika, G., Natsuaki, T., Terui, H., Kano, T., Ieki, H., Okuda, S.: "Nucleotide sequence of citrus tristeza virus seedling yellows isolate"Journal of General Plant Pathology. 67(1). 73-77 (2001)
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[Publications] Suastika,G.,Natsuaki,T.,Terui,H.,Kano,T.,Ieki,H.and Okuda,S.: "Nucleotide sequence of citrus tristeza virus seedling yellows isolate"Journal of General Plant Pathology. 67(1). 73-77 (2001)
