2006 Fiscal Year Final Research Report Summary
Studies on pathogenicity and soil-transmissibility of plant RNA viruses in the genus Furovirus
| Project/Area Number |
16380034
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Plant pathology
|
|---|
| Research Institution | The University of Tokyo |
|---|
Principal Investigator |
SHIRAKO Yukio The University of Tokyo, Asian Natural Environmental Science Center, Professor, アジア生物資源環境研究センター, 教授 (90143023)
|
|---|
| Project Period (FY) |
2004 – 2006
|
| Keywords | Genus Furovirus / Positive-sense RNA virus / Soil-borne wheat mosaic virus / Pathogenicity / Temperature sensitivity / Soil transmissibility / Yeast two-hybrid system / Chromosomal integration |
|---|
| Research Abstract |
1. Soil-borne wheat mosaic virus (SBWMV), the type species of the Genus Furovirus, forms spontaneous deletion mutations in RNA2 in infected wheat plants. Detailed analysis of deleted sites in RNA2 showed that the 3' termini of deletions end immediately upstream of the p19 promoter sequence. Deletion further into the promoter region inhibited transcription of p19 subgenomic RNA and cell-to-cell movement of viral RNA although RNA replication in single cells was not affected.
2. The p37 gene on RNA1 and CP, RT and p19 genes on RNA2 were replaced with a green fluorescence protein (GFP) gene in infectious cDNA clones of SBWMV. In vitro transcripts were inoculated to wheat leaves and GFP fluorescence in inoculated leaves and upper leaves were examined with a fluorescence light microscopy. Results showed that p37 is the viral RNA movement protein, which p19 is required for efficient cell-to-cell movement and long-distance movement of viral RNA, and that CP is required for persistent infection of the virus. CP-RT was shown to facilitate movement of viral RNA systemically.
3. The wild-type SBWMV has optimal temperatures for infection below 20℃. However, when systemically infected wheat or barley plants were kept at 25℃, virus variants that infects plants systemically at 25℃ appeared in limited plants. When the variant genome was thoroughly analyzed, only one asynonymous nucleotide change was found in p37 movement protein gene. The mutation in p37 was confirmed to be responsible for persistent infection of SBWMV variants at 25℃.
4. Increased virulence caused by mutants of SBWMV was mapped and identified in p19 gene on RNA2.
5. A conventional yeast two-hybrid system was modified by integration of BD:bait genes into yeast chromosome. Efficiency to get true-positive clones from cDNA library screening was significantly increased.
|
Research Products
(4 results)
-
-
[Journal Article] RNase E maintenance of proper FtsZ/FtsA ratio required for nonfilamentous growth of Escherichia coli cells but not for colony-forming ability.2006
Author(s)
Tamura, M., Lee, K., Miller, C.A., Moore, C.J., Shirako, Y., Kobayashi, M., Cohen, S.N.
-
Journal Title
Journal of Bacteriology 188
Pages: 5145-5152
Description
「研究成果報告書概要(和文)」より
-
-
