不同染料标识木栓细胞壁中木栓脂的分布

Distribution of Suberin in the Phellem Cells Determined by Different Dyes

以日本晚樱(Cerasus serrulata)为试验对象,取其新鲜树皮经固定、脱水、不同树脂包埋、切片,使用甲苯胺蓝、盐酸-间苯三酚、苏丹Ⅲ、苏丹红7B、硫酸氢黄连素-苯胺蓝5种染色对切片进行染色,研究木栓细胞中木栓脂的分布。结果表明:1)经不同染料染色后在明场下观察,苏丹Ⅲ的染色效果最好,可直接将木栓脂染成红色。结合甲苯胺蓝、苏丹红7B和盐酸-间苯三酚染色观察,均可确定木栓脂主要存在于木栓细胞壁的最外层。2)甲苯胺蓝可抑制木质素自发荧光,避免木质素自发荧光的干扰,直接观察到木栓脂的荧光。3)硫酸氢黄连素-苯胺蓝染色后,木栓脂的荧光在木栓细胞壁外侧呈灰绿色,木质素的荧光在木栓细胞壁的内侧呈黄绿色。4)盐酸-间苯三酚染色后,木栓细胞壁内侧富含木质素的壁层被染为红棕色,且环氧树脂包埋会降低盐酸-间苯三酚的染色效果。

In this paper, the fresh bark of Cerasus serrulata was collected, fixed with FAA solution, dehydrated, resin embedded and sliced.Then the sections were stained with toluidine blue, hydrochloric acid-resorcinol, Sudan III,Sudan red 7 B,Berberine hemisulfate salt-aniline blue to study the distribution of suberin in phellem's cell wall.The results showed that 1) after dyed and observed in bright field, Sudan III had the best dyeing effect, and it could directly dye the suberin red.Simultaneously, toluidine blue O,Sudan red 7 B and hydrochloric acid-resorcinol staining confirmed that the suberin was mainly present in the outermost layer of the phellem cell wall.2) Toluidine blue O could inhibit the auto-fluorescence of lignin, avoid the interference of the auto-fluorescence of lignin, and directly observe the fluorescence of suberin.3) After berberine bisulfate-aniline blue stained, the fluorescence of the suberin was gray-green outside the phellem cell wall, and the fluorescence of lignin was yellow-green on the inside of the phellem cell wall.4) After hydrochloric acid-resorcinol stained, the lignin inside the phellem cell wall appeared reddish brown, and the epoxy resin embedding could reduce the staining effect of hydrochloric acid-resorcinol.