Molecular Role of Calcium and Boron for Pollen Germination of Higher Plants

钙和硼对高等植物花粉萌发的分子作用

基本信息

批准号：
01560075
负责人：
SEKIYA Jiro
金额：
$ 1.28万
依托单位：
Okayama University, Faculty of Agriculture
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

项目摘要

Culture conditions for pollen germination of Lilium longiflorum on a synthetic medium were established: 1 mM CaCl_2, 1mM H_3BO_3 and 0.3 M sucrose were required for an agar medium, 0.1 mM Ca(NO_3)_2, 0.5mM H_3BO_3 and 0.3 M sucrose for a liquid medium, with a pollen density of 0-3 mg/ml. Microanalyses of calcium and boron using X-ray energy spectrometer were not successful because of difficulty in pollen tube sample preparation.EDTA and EGTA, a chelator, and H-7 as protein kinase C inhibitor, stimulated pollen germination at the early stage of germination, whereas protein synthesis inhibitor cycloheximide (CHI) inhibited the germination. However, treatment of pollen with CHI followed by EDTA treatment stimulated the germination rather than inhibited it. This suggests that protein actions required for the germination can be replaced with EDTA and that calcium may interact with cell wall and/or plasma membrane.Fatty acid composition of phosphatidylinositol (PI) changed during the pollen germination, distribution of unsaturated fatty acids such as 18:2 and 18:3 reduced while percentage of 16:0 increased. This change with PI was different from that of phosphatidylcholine. Although detection of inositol triphsphate (IP_3) has not been successful in the pollen germination system in our experiment, the above result suggests that PI-mediated calcium regulation system may play an important role during pollination.To examine calcium effect on cell wall loosening, cell wall degrading enzgme activities were assayed. Significant activities of polygalacturonase and beta-1, 3-glucanase were detected. However, addition of calcium to the medium neither stimulated nor inhibited these enzyme activitie. Now we successfully purified beta-1, 3-glucanase from L. longiflorum leaves to administrate to rabbit for preparation of anti-beta-1, 3-glucanase serum and immuno-histological investigation of pollen termination in connection with calcium and boron action.

建立了在合成培养基上的lium limifforum粉的培养条件：1 mm CaCl_2、1MM H_3BO_3和0.3 M蔗糖对于琼脂培养基需要0.1 mm Ca（NO_3）_2、0.5mm H_3BO_3和0.3bo_3和0.3 M含量，用于液体中等含量的液体含量为0-3 mg，液体含量为0-3 mg。 Microanalyses of calcium and boron using X-ray energy spectrometer were not successful because of difficulty in pollen tube sample preparation.EDTA and EGTA, a chelator, and H-7 as protein kinase C inhibitor, stimulated pollen germination at the early stage of germination, whereas protein synthesis inhibitor cycloheximide (CHI) inhibited the germination.但是，用CHI进行花粉的处理，然后进行EDTA处理刺激了发芽，而不是抑制它。这表明发芽所需的蛋白质作用可以用EDTA代替，并且钙可能与细胞壁和/或质膜相互作用。磷脂酰肌醇（PI）的致命酸组成在花粉发芽期间发生了变化，在不饱和脂肪酸的分布中，如18：2和18：3的分布（如18：2和18：3）减少了16：0的百分比。 PI的这种变化与磷脂酰胆碱的变化不同。尽管在我们的实验中，在花粉发芽系统中检测肌醇三氯（IP_3）尚未成功，但上述结果表明，PI介导的钙调节系统可能在授粉过程中起重要作用。检查钙对细胞壁松动的钙效应，测定了细胞壁降解的细胞壁降解活性。检测到聚半乳糖苷酶和β-1，3-葡聚糖酶的显着活性。但是，在培养基中添加钙既没有刺激也不抑制这些酶。现在，我们成功地纯化了从长叶叶乳杆菌的3-葡聚糖酶到兔给兔子的3-葡聚糖酶，以制备抗β-1，3-葡萄糖酶血清以及与钙和硼作用有关的花粉终止的免疫历史研究。