| Project/Area Number |
14560036
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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 |
OKUDA Seiichi Utsunomiya University, Faculty of Agriculture, Professor, 農学部, 教授 (90091941)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2004: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2003: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Crinivirus / Closterovirus / whitefly / PCR / Cucumber yellows virus / Tomato infectious chlorosis virus / dsRNA / 抗血清 |
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| Research Abstract |
Molecular cloning and sequencing of plant virus genomes has been carried out during the past decade. One objective of molecular analysis is improvement of the methods to detect and diagnose pathogenic viruses. For molecular analysis, viral nucleic acids are usually extracted from purified virus preparations. However, there are many recalcitrant viruses that are unable to purify by current methods and, therefore, the viral nucleic acid samples and their antisera are not available. Cucumber yellows virus(CuYV) is one of such recalcitrant viruses. In this research, the application of dsRNA extraction technique from plants was applied to get viral RNA, replicative form of dsRNA(RF-dsRNA). The first objective of this research was the production of cDNA clones of CuYV and complete nucleotide sequence of CuYV genome was determined. This is the first record in the world and suggests that CuYV is a member of the genus Crinivirus. Occurrence of CuYV was also detected in Indonesia but not in Pakistan. However, the virus could not be detected from its whitefly vector by PCR. In this research Tomato infectious chlorosis virus, a member of criniviruses, was newly recognized in Japan. We have also attempted that the dsRNA extraction technique was used to construct a plasmid to express the viral coat protein in E.coli. Finally production of the antiserum against carrot cryptic virus was succeeded, indicating that this method is useful to prepare antisera against ciriniviruses for which no antisera are available in present.
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