
竹叶去硅方法的筛选及其细胞学研究
杨雅运, 傅华君, 苏佳露, 万雅雯, 张春霞, 丁雨龙, 林树燕
南京林业大学学报(自然科学版) ›› 2019, Vol. 43 ›› Issue (5) : 74-80.
竹叶去硅方法的筛选及其细胞学研究
Desilication of bamboo leaves for paraffin slices preparation and their cytological study on eleven bamboo species
【目的】竹类植物是我国重要的经济植物资源,竹叶表皮细胞富含硅质,导致制片过程中切片易破碎,不能获得清晰完整的竹叶切片。开发竹叶切片处理方法,对于研究竹类植物叶片解剖结构具有重要的实践和应用价值。筛选有效的氢氟酸处理浓度以及时间是观察竹叶内部解剖结构的前提。【方法】采用石蜡制片的方法,以竹叶为材料,首先优化去硅环节,筛选出适用于竹叶的最佳氢氟酸浓度和时间;之后采用此最佳处理方法制片,观察竹叶内部细胞结构并结合去硅效果进行分析。【结果】通过对竹叶叶表皮和基本组织的形态解剖结构进行比较,发现真正影响竹叶完整石蜡切片制备的结构是叶片上下表皮的硅质细胞和主脉,这些结构使得制片易破碎,无法得到完整竹叶内部解剖结构。通过试验发现经过体积分数为25%的氢氟酸去硅处理2 d后再进行石蜡制片可以使竹叶上下表皮细胞和叶脉结构完整,得到理想的石蜡切片。【结论】竹叶去硅处理最佳氢氟酸体积分数为25%,处理时间以2 d为宜。
【Objective】 Bamboo is an important economic plant in China. As the epidermis cells of bamboo leaves are rich in silicon, paraffin slices of bamboo epidermal cells are easily broken and thus rarely provide a clear and complete sample. Nevertheless, the paraffin method for preparation of bamboo leaf sample has important practical applications in the study of bamboo leaf anatomical structure. Therefore, it is very necessary to desiliconize with hydrofluoric acid before paraffin sectioning. Determining the optimum concentration of hydrofluoric acid for treatment and treatment duration is crucial for observing the internal anatomical structure of bamboo leaves. 【Method】 In this study, we used the paraffin method to prepare leaf samples of 11 bamboo species. First, we determined the optimum hydrofluoric acid concentration and treatment duration for the desilication of bamboo leaves; the other 10 bamboo species were then subjected to this treatment. The bamboo leaf cell structure as well as the effect of desilication were observed and analyzed. 【Result】 By comparing the morphological and anatomical structures of the leaf epidermis with those of leaf basic tissues, the structures that markedly affected the quality of the paraffin slice were determined to be the siliceous cells of the upper and lower epidermis and the main vein. These structures render the traditional paraffin slice easily breakable and make it difficult to obtain a paraffin slice of the whole internal anatomical structure of bamboo leaves. In this experiment, we found that with desilication of the upper and lower epidermal cells and veins of bamboo leaves using 25% hydrofluoric acid for 2 days resulted in the successful preparation of complete paraffin sections. 【Conclusion】 The optimum hydrofluoric acid concentration for desilication is 25% and the treatment duration is 2 days.
bamboo leaves / paraffin section / desiliconization / anatomical structure
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