Fluorescence labeling method for cell DNA during cotton pollen development process

Abstract

The invention discloses a fluorescence labeling method for detecting DNA matters of a cotton pollen mother cell, and belongs to the biological technical field. The method comprises the following steps: on a sunny morning, buds are picked up, and a sample passes through ethanol with reducing concentrations to enter distilled water; pollen mother cells with a diameter of less than 7mm of the buds are placed in glycerin for infiltration for 2 to 3 hours, and, after washing, are dyed by DAPI in the dark for 1 to 2 hours to label DNA matters in the cells; mature pollen grains with the diameter of more than 7mm of the buds are added with a 10 percent sodium hypochlorite aqueous solution, and subjected to water bath treatment at a temperature of 60 DEG C for 10 minutes, washed, placed in 0.1 to 0.2 percent glycerin for infiltration for 2 to 3 hours, washed, added with DAPI solution, stood for 10 minutes and subjected to microscopic examination to label DNA matters in the cells. The method has the advantages of simple and quick operation, less damages to cells, good labeling effect and easy observation, and can play an important role in research on the development process of cotton male gametophyte, and has high application value.

Description

1. Technical field [0001] The invention relates to a fluorescent labeling method for DNA material of cotton pollen mother cells, belonging to the field of biotechnology. 2. Background technology [0002] The change process from plant microsporogenesis to pollen mother cell maturity has always been one of the important contents of reproductive biology research. The research on cotton microsporogenesis and male gamete development began in the 20th century. The early research methods were mainly paraffin section method, stained with hematoxylin, safranin, and fast green, and then developed into ultrathin section and electron microscope observation. Or in vitro culture, fluorescent staining. But until now, due to the complicated operation of these methods, it is difficult to quickly observe the whole process of male gametophyte development, and the observation effect is not ideal. There are two main reasons for this, one is the limitation of the experimental material itself; t...

AI Technical Summary

Problems solved by technology

In addition, the specificity of the dyes used by the predecessors to detect the development of cotton male gametophytes is not strong, and the experimental procedures are relatively cumbersome and the cycle is long.

So there is no good solution yet

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