葡萄种子细胞中脱落酸的胶体金免疫电镜定位

Immunogold Electronic-microscopy Localization of Abscisic Acid in Seed Cells of Grapvine

用胶体金免疫电镜技术，对脱落酸（ABA）在葡萄种子细胞超微结构水平上的分市进行了研究。按葡萄种子细胞液泡中是否含有电子密度大的染色物质，可将其分为含酚和无酚2个主要类型。在含酚细胞中电子密度大的染色物质呈多种状态存在，先是均匀而稀疏地分布在液池中，然后逐渐凝聚成块状，最后浓缩成一薄层并均匀分布在液泡膜内侧。无酚细胞中金颗粒主要标记在细胞核和细胞质，特别是细胞核有大量金颗粒标记，细胞壁有少量金颗粒标记，液泡中没有发现金颗粒标记。和无酷细胞相似，在含酚细胞中金颗粒主要标记在细胞核和细胞质，特别是细胞核金颗粒密度很大，细胞壁有极少金颗粒标记。引人注目的是液泡中聚集态的染色物质上也发现金颗粒标记，但稀疏而均匀分布的染色物质上几乎找不到金颗粒标记，当染色物质最终浓缩成薄层状态时则发现有大量的金颗粒标记。无论用免疫前兑血清染色的对照切片还是材料，不被EDC固定的对照切片中都很难找到金颗粒标记，说明该ABA免疫胶体金电镜定位结果是特异、可靠的。

Abscisic acid (ABA) localization was carried out with immunogold electronmicroscopy technique in seed cells of grapevine (Vitis vindera L. ×Vitis labrusca L. ). Two different kinds of cells were observed in the grape seed, one kind containing in their vacuole large amounts of electronically dense substances (phenolic compounds?) (called phenol-rich cells ), and another containing no such substances (called non-phenol cells). In the phenolrich cells the electronically dense substances appeared in many different states, from the sparsely and evenly distributed state gradually aggregating to lumped state and finally concentrating to a thin and very dense layer near the vacuole membrane. In the non-phenol cells both nucleus and cytoplasm were heavily labeled by gold particles, and few gold particles were also found in the cell wall, but no particles were observed in the vacuole. In the phenolrich cells both nucleus and cytoplasm were heavily labeled by gold particles, and few particles were also found in cell wall, which is similar to the non-phenol cells. In the phenol-rich cells the electronically dense substances-distributed areas were also labeled by gold particles specially when these substances aggregated to the lumped and the final thin layer state.Gold particles were scarcely found nor in the sections treated with pre-immune rabbit serum instead of the anti-ABA IgG neither in the sections without EDC fixation, which demonstrated that the immunogold localization of ABA was specific and reliable.