一种改进整体染色和透明技术观察棉花珠心的方法

An Improved Method of the Whole Staining and Clearing Technique to Ob-serve the Nucellus of Cotton

棉花花器官较大,较厚的珠被组织导致激光共聚焦等现代显微技术难以直接观察其受精过程,影响了其生殖发育相关方面的研究进展。为了观察棉花的生殖发育过程,本研究通过剥离珠心进行整体染色和透明实现了对棉花受精过程的观察,使用不同浓度天青C对棉花珠心进行染色,经水杨酸甲酯(BM)透明化处理观察棉花生殖细胞。结果表明,该方法能够清晰观察到棉花雌配子体胚囊内各细胞的细胞膜、细胞核、核膜、核仁等显微结构。本研究仅需简单的实验器材与少量的人工操作即可对棉花这类胚囊组织较大的植物进行细胞观察,同时为研究棉花生殖发育提供了一种简便快捷的显微观察方法。

Cotton has a larger floral organ.However,due to the thick bead structure,modern microscopes,such as laser confocal microscopy,are difficult to observe the specific process of female gametophyte directly,which af-fects its research progress.To observe the process of the cotton reproductive development process,this study ob-served the fertilization process of cotton by stripping the nucellus for whole staining and transparency.Cotton nu-cellus were stained with Azure C of different concentrations,and the nucellar tissue was clearly treated with Birch-Me(BM)to observe female gametophyte.The results showed that this method can clearly distinguish the microstructures of the cell membrane,nucleus,and nuclear membrane.This method only needed simple experi-mental equipment and a small amount of manual operation to observe the cell of cotton and other plants such as cotton for large embryo sac tissue.,At the same time,it provides a simple and effective method for the study of cotton reproductive development.