| Project/Area Number |
14360205
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Applied molecular and cellular biology
|
|---|
| Research Institution | Yokohama National University |
|---|
Principal Investigator |
HIRATSUKA Kazuyuki Yokohama National University, Faculty of Environment and Information Sciences, Professor, 大学院・環境情報研究院, 教授 (30202279)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
NAKAMURA Tatsuo Yokohama National University, Faculty of Environment and Information Sciences, Lecturer, 大学院・環境情報研究院, 講師 (50334636)
SUZUKI Masashi Yokohama National University, Faculty of Environment and Information Sciences, Associate Professor, 大学院・環境情報研究院, 助教授 (40282694)
|
|---|
| Project Period (FY) |
2002 – 2003
|
| Project Status |
Completed (Fiscal Year 2003)
|
| Budget Amount *help |
¥11,500,000 (Direct Cost: ¥11,500,000)
Fiscal Year 2003: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥7,900,000 (Direct Cost: ¥7,900,000)
|
|---|
| Keywords | reproduction / meiosis / large-scale analysis / nuclear localization signal / regulated gene expression / transgenic plants / trumpet lily / Arabidopsis / アンチセンス法 |
|---|
| Research Abstract |
In order to study gene functions in plant cells we developed particle gene delivery system for foreign gene expression in various plant cells. We chose to use Lilium longiflorum as model system because no reliable transformation method is available for lilies. Using lily cDNA library generated from male reproductive organs at various developmental stages, we examined the functional genes involved in microsporogenesis by large-scale sequencing and obtained more than 1000 ESTs. Based on the results obtained by the reverse-northern analysis, we selected number of cDNA clones for further analysis. A cDNA clone, designated M532 showed no extensive sequence homology to previously characterized gene products and specifically expressed during microsporogenesis. Interestingly, the M532 protein targeted to the nucleolus when fused to GFP. Another cDNA clone encoding a protein, designated LISCL, exhibited high similar to the GRAS family. A series of transcriptional activation experiments of truncated LISCL proteins fused to the yeast GAL4 DNA-binding domain clearly demonstrated that the amino terminus of the LISCL protein has a transcriptional activity of the meiosis-associated promoter revealed that in pollen mother cells of lily, the activity of the meiosis-associated promoter is specifically enhanced by the LISCL protin co-expression. These results suggest that the LISCL is involved in the transcriptional regulation during microsporogenesis within the lily anther. We also isolated viral cDNAs from reproductive organs and obtained a full-length cDNA. Sequence analysis revealed that the virus shows strong similarity to the lily symptomless virus. Currently, we are in the process of developing a gene vector system using the full-length infectious clone of the viral cDNA.
|
