| Project/Area Number |
23K14210
|
|---|
| Research Category |
Grant-in-Aid for Early-Career Scientists
|
| Allocation Type | Multi-year Fund |
|---|
| Review Section |
Basic Section 44030:Plant molecular biology and physiology-related
|
|---|
| Research Institution | Nagoya University |
|---|
Principal Investigator |
Kozgunova Elena 名古屋大学, 高等研究院(理), 特任助教 (90786120)
|
|---|
| Project Period (FY) |
2023-04-01 – 2027-03-31
|
| Project Status |
Granted (Fiscal Year 2023)
|
| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2026: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2025: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2024: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2023: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
|
|---|
| Keywords | chloroplast division / Physcomitrium patens / cell division / 葉緑体 |
|---|
| Outline of Research at the Start |
葉緑体は、植物が太陽からエネルギーを得て炭水化物に変換する「光合成」を行うために必要な細胞です。また、葉緑体は、植物体の成長に伴い、植物細胞内で増殖していきます。葉緑体の分裂過程は、分子レベルではまだ十分に解明されていません。本研究では、最近発見されたTHIOLというタンパク質の葉緑体分裂における機能を明らかにすることを目的としています。P. patensとA. thalianaという2つの異なる植物モデル系を用いて、遺伝子編集、ライブセルイメージング、高解像度顕微鏡、in vitroアッセイなど、様々な分子生物学の手法を用いて、植物系統を通じたTHIOLの機能を研究します。
|
| Outline of Annual Research Achievements |
This research aims to characterize a novel chloroplast division gene THIOL, using two model plant: Physcomitrium patens and Arabidopsis thalinan. In P. patens framshift mutations in THIOL gene result in a severe phenotype with dwarf leaves and few large chloroplasts per cell. We generated frameshift mutations in the THIOL homologue in A.thaliana and observed a similar phenotype with tiny leaves and large chloroplasts. These results suggest that THIOL gene function is conserved across multiple plant species.
We also generated two fluorescent marker lines for known chloroplast division genes: FtsZ and ARC5. Chloroplasts divide by forming two contractile rings: inner ring consisting of FtsZ and outer ring, formed by ARC5. Based on detailed observation of chloroplast division, contraction or shrinkage of the chloroplast division ring is affected after THIOL mutation, preventing complete separation of dividing chloroplasts. These results provide possible explanation of THIOL frameshift phenotype.
Next, we will perform further analysis to characterize THIOL role in plant cells.
|
| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
Funstional analysis of THIOL gene have been progressing well. We succesfully generated frameshift mutant in A. thaliana to supplement our previous results obtained from P. patens. In addition, we gained further insights into chloroplast division defects in THIOL mutant.
On the other hand, localization analysis of THIOL gene so far was not succesful. THIOL gene codes a small protein, traditional methods to observe localization such as GFP fusion did not work, as the protein was non-functional. Next, we will try split-GFP system to analyze THIOL localization.
|
| Strategy for Future Research Activity |
In future, we will perform rescue and cross-rescue experiments to validate frameshift mutant phenotype. We also plan to investigate THIOL interacting partners, but performing proximity labelling experiments, such as TURBID, followed by mass-spectrometry. It will allow us to identify THIOL protein network and binding partners.
Our main focus is to show THIOL localizatio inside the cell. We will try a split-GFP system, which often works better comparing to original GFP for small proteins. If it split-GFP system does not work, we will perform immunostaining experiments using antibody against THIOL.
|
