Dyeing method for observing anther development structure changes of plants

Abstract

The invention relates to a dyeing method for observing anther development structure changes of plants. By use of the dyeing method, the problem of clearly observing of an anther wall tapetum and structure changes and an anther wall can be effectively solved. The dyeing method comprises the following steps: fixing an anther by using a mixed solution containing formaldehyde, glacial acetic acid and alcohol, performing dehydration by using alcohol, then treating the anther for three times in sequence in a mixed solution containing n-butyl alcohol and alcohol, and soaking the anther in n-butyl alcohol; then transferring the anther into a mixed solution containing n-butyl alcohol and paraffin, pouring out half of the mixed solution and adding an equal weight of paraffin every 3h, and finally, pouring out the mixed solution, embedding the anther into a paraffin block by using paraffin, and cutting the paraffin block into a paraffin belt; and coating gelatin on a glass slide, attaching the paraffin belt to the glass slide, baking the paraffin belt, performing dewaxing, then treating the glass slide once in alcohol and distilled water respectively, treating the glass slide with the anther in periodic acid, rinsing the glass slide by using distilled water, performing dyeing in a Schiff reagent, rinsing the glass slide by using distilled water, and covering the glass slide with another glass slide. The dyeing method provided by the invention is easy to operate, good in dyeing effect, and clear and accurate in observation of the anther development structure changes of the plants.

Description

technical field [0001] The invention relates to plant tissues, in particular to a dyeing method for observing the development and structure changes of plant anthers. Background technique [0002] The research on anther development and structure has always been a hotspot in the research of reproductive biology. In anthers, the nutrients required for the pollen mother cell meiosis to form pollen and the sporopollenin constituting the outer wall of the pollen are secreted by the velvet cells of the anther wall or produced by the disintegration of the velvet cells. Afterwards, the velvet layer disappears; thus, the proper timing of the formation and disintegration of the velvet layer is very important for the normal development of pollen. In addition, in the study of anther development, it is very necessary to clearly observe the formation and disappearance time of the velvet layer, the structure change time of the pollen wall and the relationship between the two changes in tim...

AI Technical Summary

Problems solved by technology

In the past, staining of paraffin sections was often used for the research on pollen development and structure. For example, aniline blue can stain various macromolecules in cells to display different colors, while polysaccharide / starch, lipid and protein staining are stained with periodate respectively. -Schiff's reagent (PAS), Sudan black B and Coomassie brilliant blue staining, all of these staining methods are observed under bright field with a microscope, due to the complex structure of the anther wall (four-layer structure including the velvet layer) , the above-mentioned anther staining and observation methods often cannot clearly observe the structural changes of the anther wall and the anther wall

[0003] In the previous studies on the development of anther structure, the PAS method was often used for staining, but the PAS method is mainly used for staining insoluble polysaccharides, so that the cell walls, pollen and starch grains of each layer of the anther are dyed red. Under bright field observation, the structural changes of the anther wall and the anther wall cannot be clearly observed