Pollen-pistil interaction in Brassicaceae

十字花科花粉与雌蕊的相互作用

• 批准号: 17570037
• 负责人: IWANO Megumi
• 金额: $ 2.18万
• 依托单位: Nara Institute of Science and Technolohy
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17570037/
• 关键词: actin; Arabidopsis; calcium; FRET; imaging; pollination; yellow cameleon; アブラナ科植物; 授粉; Caイオン; 突然変異体

1. We expressed yellow cameleon 3.6, a Ca^<2+> indicator based on green fluorescent protein (GFP), in the pollen grains and papilla cells of Arabidopsis thaliana and monitored [Ca^<2+>]_cyt dynamics during pollination. In the pollen grain, [Ca^<2+>]_cyt, increased at the potential germination site soon after hydration and remained augmented until germination. As in previous in vitro germination studies, [Ca^<2+>]_cyt oscillations were observed in the tip region of the growing pollen tube, but the oscillation frequency was faster and [Ca^<2+>]_cyt was higher than had been observed in vitro. In the pollinated papilla cell, remarkable increases in [Ca^<2+>]_cyt occurred three times in succession, just under the site of pollen-grain attachment. [Ca^<2+>]_cyt increased first soon after pollen hydration, with a second increase occurring after pollen protrusion. The third and most remarkable [Ca^<2+>]_cyt increase took place when the pollen tube penetrated into the papilla cell wall.2. We exa … More mined the role of actin, a key cytoskeletal component regulating the transport system for hydration and germination, in the papilla cell during cross-pollination. Using rhodamine-phalloidin staining, we showed that cross-pollination induced actin polymerization. By monitoring transiently expressed GFP-mTalin, we observed the concentration of actin bundles at the cross-pollen attachment site and actin reorganization. We further showed that the coat of cross-pollen is sufficient to mediate this response. The actin-depolymerizing drug cytochalasin D (CD) significantly inhibited pollen hydration and germination during cross-pollination, further emphasizing a role for actin in these processes. Additionally, three-dimensional electron microscopic tomography revealed the close association of the actin cytoskeleton with an apical vacuole network. Cross-pollination led to vacuolar rearrangements toward the site of pollen attachment. Taken together, our data suggest that cross-pollination differentially affect the dynamics of actin cytoskeleton leading to changes in vacuolar structure associated with hydration and germination. Less

1.我们在拟南芥的花粉粒和乳头细胞中表达了基于绿色荧光蛋白（GFP）的Ca^<2+>指示剂黄茶3.6，并监测授粉期间的[Ca^<2+>]_cyt动态。在花粉粒中，[Ca 2+ ]_cyt在水合后不久在潜在的发芽位点增加，并且保持增加直至发芽。与之前的体外萌发研究一样，在生长的花粉管的尖端区域观察到[Ca^<2+>]_cyt振荡，但振荡频率更快，并且[Ca^<2+>]_cyt高于体外观察到的。在授粉的乳头细胞中，[Ca^2+]_cyt连续3次显着增加，就在花粉粒附着位点下方。 [Ca^2+]_cyt在花粉水合后不久首先增加，在花粉突出后发生第二次增加。第三次也是最显着的[Ca^2+]_cyt增加发生在花粉管穿入乳头细胞壁时。2．我们挖掘了肌动蛋白的作用，肌动蛋白是异花授粉过程中乳头细胞中调节水合和发芽运输系统的关键细胞骨架成分。使用罗丹明-鬼笔环肽染色，我们发现异花授粉诱导肌动蛋白聚合。通过监测瞬时表达的 GFP-mTalin，我们观察了交叉花粉附着位点肌动蛋白束的浓度和肌动蛋白重组。我们进一步表明，交叉花粉的外壳足以介导这种反应。肌动蛋白解聚药物细胞松弛素 D (CD) 显着抑制异花授粉过程中的花粉水合和发芽，进一步强调了肌动蛋白在这些过程中的作用。此外，三维电子显微镜断层扫描揭示了肌动蛋白细胞骨架与顶端液泡网络的密切联系。异花授粉导致液泡向花粉附着部位重新排列。综上所述，我们的数据表明，异花授粉对肌动蛋白细胞骨架的动力学有不同的影响，导致与水合和发芽相关的液泡结构的变化。较少的

项目成果

Synthesis of Rubisco gene products is upregulated by increasing the copy number of rbcL gene in Chlamydomonas chloroplast genome, without increased accumulation of the two Rubisco subunits.

通过增加衣藻叶绿体基因组中 rbcL 基因的拷贝数来上调 Rubisco 基因产物的合成，而不增加两个 Rubisco 亚基的积累。

• 发表时间: 2005
• 期刊: Plant Biotech. 22
• 作者: Jang;M. -H.;高村典子;Takamura N.;高村典子;H.Shimosato et al.;M.Iwano et al.;M.Iwano et al.;H.Shimosato;M.Iwano;M.Iwano;H.Shimosato;M.Iwano;H.Shiba et al.;H.Shiba;M.Iwano;H.Shiba;Y.Yoshie et al.;H.Uchida et al.
• 通讯作者: H.Uchida et al.

Dominance relationships between self-incompatibility alleles controlled by DNA methylation

• DOI: 10.1038/ng1734
• 发表时间: 2006-03-01
• 期刊: NATURE GENETICS
• 影响因子: 30.8
• 作者: Shiba, H;Kakizaki, T;Takayama, S
• 通讯作者: Takayama, S

Concurrent occurrence of respiratory activity in the thylakoidal membrane of a cyanobacterium, Nostoc muscorum as revealed by quantitative histochemistry.

定量组织化学揭示蓝藻发菜的类囊体膜同时发生呼吸活动。

• 发表时间: 2007
• 期刊: J. Electr. Microsc. Technol. Med. Biol. (in press)
• 作者: Jang;M. -H.;高村典子;Takamura N.;高村典子;H.Shimosato et al.;M.Iwano et al.;M.Iwano et al.
• 通讯作者: M.Iwano et al.

Actin dynamics in papilla cells of Brassica rapa during self- and cross-pollination

• DOI: 10.1104/pp.106.095273
• 发表时间: 2007-05-01
• 期刊: PLANT PHYSIOLOGY
• 影响因子: 7.4
• 作者: Iwano, Megumi;Shiba, Hiroshi;Takayama, Seiji
• 通讯作者: Takayama, Seiji

FRET analysis in plant cells

植物细胞中的 FRET 分析

• 发表时间: 2006
• 期刊: Protocol to observe plant cells (Eds. H.Fukuda, M.Nishimura and A.Nakano) (SHUJYUNSHA)
• 作者: Jang;M. -H.;高村典子;Takamura N.;高村典子;H.Shimosato et al.;M.Iwano et al.;M.Iwano et al.;H.Shimosato;M.Iwano;M.Iwano;H.Shimosato;M.Iwano;H.Shiba et al.;H.Shiba;M.Iwano
• 通讯作者: M.Iwano