陈利娜 薛辉 李好先

摘要[目的]摸索适合石榴花芽及胚珠结构观察的石蜡切片方法。[方法]以“突尼斯软籽”石榴花芽及胚珠为试材，在传统石蜡切片方法的基础上，结合石榴花芽鳞片厚、子房多室的特点，对样品处理、软化、透明、浸蜡、染色等重要制片环节进行研究。[结果]不同样品处理方式对比发现，将子房纵切4瓣进行固定，切片效果较好。软化结果表明，纱布包裹花芽浸入水中加热至沸腾30 min，有助于提高制片效果。不同染色方式对比发现，爱氏苏木精染色法更适合石榴胚珠切片观察。[结论]通过优化常规石蜡切片方法，得到了适合石榴花芽及胚珠结构观察的石蜡切片方法，该方法同样适用于与石榴花芽及胚珠结构类似的其他结构。

关键词 石榴；花芽；胚珠；石蜡切片

中图分类号 S665.4 文献标识码 A 文章编号 0517-6611（2017）02-0001-03

Abstract[Objective] To grope the suitable paraffin section method for pomegranate floral bud and ovule.[Method] The floral buds and ovules of Tunisiruanzi were used as experimental materials for paraffin sectioning to improve sample treatment.Given the character of pomegranate floral with thick bud scales and ovary of multicellular， the process of softening， clearing， waxing and staining were improved.[Result] To confirm the optimal fixation methods for different parts of ovary， we compared the different fixation methods and found that the best method is fixing after cutting the ovary into four parts using F.A.A..Softening results showed that if the floral bud was boiled for 30 minutes without stripping scales， the tissues would be complete.Paraffin section staining was performed with Ehrlich hematoxylin， which was the best material for staining for pomegranate floral bud and ovule.[Conclusion] A modified method of paraffin sectioning for pomegranate floral bud and ovule was recommended on the bases of the traditional method.

Key words Pomegranate；Floral bud；Ovule；Paraffin section

石榴（Punica granatum L.），為石榴科石榴属果树，在我国广泛栽培，且栽培历史悠久。石榴用途广泛、形态美观。石榴加工产品对癌症、冠状动脉心脏疾病、动脉粥样硬化、高胆固醇血症、高脂血症、高血压、传染性疾病等具有显著疗效[1]。石榴花期长、花量大，但成花过程中发生严重的雌蕊败育，形成大量败育花，败育花的比率与坐果率呈负相关，是落花落果、坐果率低的主要原因[2]。Wetzstein等[3]对石榴2种不同类型花形态进行比较，发现石榴败育花的形成与其胚珠发育息息相关。因此，对石榴花芽及胚珠胚胎学进行研究尤为重要。

石蜡切片是胚胎学研究中常用的一种技术，作为一种辅助研究技术，广泛应用于植物花芽分化等研究[4-12]。……