Molecular-cytological analysis of plant fertillization and early-embryogenesis by using in vitro fertilization system and micromanipulation techniques

利用体外受精系统和显微操作技术对植物受精和早期胚胎发生进行分子细胞学分析

• 批准号: 14540589
• 负责人: HIGASHIYAMA Tetsuya
• 金额: $ 2.62万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14540589/
• 关键词: plant; fertilization; pollen tube; sperm cell; embryo sac; synergid cell; attractant; kinetics; 胚嚢; 花粉管誘引物質; 重複受精; 顕微分子細胞学的解析; 体外受精; トレニア

Species specificity in the attraction signal of the pollen tube was found by using three species of Torenia and two species of Vandellia. In these plants, the embryo sac protrudes from the ovule and it was confirmed that the attraction signal is derived from the synergid cell by laser ablation. When ovules of Torenia were cultivated with those of other species, pollen tubes of each species tended to grow toward the embryo sac of the same species. It was also found that the species specificity might be a cause of reproductive barrier in vivo. Consistently with the species specificity, previous candidates for the attractant such as Ca^<2+>and GABA were shown not to be the attractant from the synergid cell. It was suggested that the attractant might be some molecules synthesized in the synergid cell.I also developed a visualization system of living gametophytic cells. By staining with chemical fluorescent probes and using the a sensitive camera system, behavior and kinetics of two sperm cells was revealed. Trnasformation system to use GFP was also prepared. When a GFP gene fused with ACT11 promoter of Arabidopsis was delivered with bombardment, bright fluorescence was observed in the embryo sac of Torenia. these techniques established here would be important for future works.

通过使用三种多伦尼亚和两种范德利亚种，发现花粉管的吸引信号中的物种特异性。在这些植物中，胚胎囊从胚珠突出，并证实吸引信号是通过激光消融源自相关细胞的。当将多雷尼亚的胚珠与其他物种的胚珠一起培养时，每种物种的花粉管倾向于生长在同一物种的胚胎囊中。还发现该物种特异性可能是体内生殖屏障的原因。与物种特异性一致，以前的吸引剂（例如Ca^<2+>和GABA）的候选者并非是同性细胞中的吸引剂。有人提出，吸引剂可能是在协同细胞中合成的某些分子。我还开发了一种活物配子细胞的可视化系统。通过用化学荧光探针染色并使用敏感的摄像头系统染色，可以揭示两个精子细胞的行为和动力学。还准备了使用GFP的Trnasformation系统。当将拟南芥ACT11启动子融合的GFP基因被轰击传递时，在多雷尼亚的胚胎囊中观察到明亮的荧光。这些在这里建立的技术对于未来的作品至关重要。

项目成果

東山哲也: "植物の受精のしくみを解明"遺伝. 56. 20-21 (2002)

Tetsuya Higashiyama：“阐明植物受精机制”遗传学 56. 20-21 (2002)。

Yagisawa F., Mori T., Higashiyama T., Kuroiwa H., Kuroiwa T.: "Regulation of Brassica rapa chloroplast proliferation in vivo and in cultured leaf disks."Protoplasma. 222. 139-148 (2003)

Yagisawa F.，Mori T.，Higashiyama T.，Kuroiwa H.，Kuroiwa T.：“体内和培养叶盘中芸苔叶绿体增殖的调节。”原生质体。

Higashiyama T.: "Chemicals in plant fertilization"Syokubutono-Seichou Chousetsu. (in press).

Higashiyama T.：“植物施肥中的化学物质”Syokubutono-Seichou Chousetsu。

Higashiyama T.: "The synergid cell : attractor and acceptor of the pollen tube for double fertilization"Journal of Plant Research. 115. 149-160 (2002)

Higashiyama T.：“助细胞：双受精花粉管的吸引子和受体”植物研究杂志。

東山哲也: "受精のガイダンス機構"蛋白質核酸酵素. 47. 1645-1650 (2002)

Tetsuya Higashiyama：“受精的指导机制”蛋白质核酸酶。47。1645-1650（2002）