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Fluorescence labeling method for cotton pollen tube microfilament framework

A technology for fluorescent labeling and pollen tubes, applied in the biological field, can solve problems that have not been reported, and achieve high fluorescent staining efficiency and good labeling effect

Inactive Publication Date: 2009-07-08
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, due to the limitations of experimental materials and detection methods, effective methods and techniques for labeling microfilament skeletons in upland cotton powder tubes have not been reported

Method used

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  • Fluorescence labeling method for cotton pollen tube microfilament framework

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Embodiment 1

[0021] Example 1. Fluorescence labeling of Lumian 21 pollen tube microfilament skeleton and its microscopic observation

[0022] First carry out fluorescent labeling to the pollen tube of Lumian 21 (kind for production) with the method of the present invention, comprise the following steps:

[0023] 1) Punch holes on the glass slide, drill a circular hole with a diameter of 1cm in the middle of the slide glass (25.5×76.2mm, 1mm-1.2mm thick) with an electric drill, and seal one side of the hole with a cover glass, adhesive For nail polish, let it stand at room temperature until the nail polish dries, stick the cover glass and slide glass together to form a small chamber, and set aside;

[0024] 2) Prepare semi-solid pollen tube culture medium, weigh 0.01g boric acid, 0.03g calcium nitrate tetrahydrate, 0.02g magnesium sulfate heptahydrate, 15g polyethylene glycol 8000 and 20g sucrose respectively, dissolve them in deionized water and adjust the pH to 6.2, and then make the vol...

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Abstract

The invention relates to a method for carrying out fluorescence labeling on an actin filament framework of a cotton pollen tube, which belongs to the technical field of biology. The method comprises: perforating a glass slide to make a chamber for cultivating the pollen tube; cultivating pollen in the chamber to bourgeon the pollen tube; fixing the cultivated pollen tube by 50mm Pipes buffer solution (pH6.9) containing 4 percent of paraformaldehyde for 1 to 2 h; washing the pollen tube, and putting the pollen tube into 0.1 to 0.2 percent glycerin (or 0.1 to 0.2 percent Triton X-100) for 3 to 4 h at a room temperature (or over night in a 4 DEG C refrigerator); washing the pollen tube, and staining the pollen tube through a phalloidine labeled by TRITC(rhodamine) with the concentration of 1 to 2.5 mu g / ml; and washing the pollen tube, and then observing the pollen tube under a confocal scanning microspope. The method has the advantages of simple operation, good labeling effect and important practical application value, and can play an important part in researching the framework of a cotton pollen tube cell and fertilization reproduction.

Description

1. Technical field [0001] The invention relates to a fluorescent labeling method of pollen tube microfilament framework in the field of plant biotechnology, and belongs to the field of biotechnology. 2. Background technology [0002] The pollen tube is the carrier of the male reproductive unit in the fertilization process of flowering plants. A series of physiological activities such as its germination and elongation directly affect whether the fertilization process can be completed smoothly. The growth of the pollen tube is different from other plant cells. It has Polar apical growth is dependent on the normal aggregation of microfilaments in the growing pollen tube. Therefore, using the pollen tube system to study the microfilament skeleton and its function in organisms is one of the most active research fields at present, and it is also the research focus in the field of plant reproductive biology. Studies in recent years have shown that microfilaments are an important p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
Inventor 唐灿明岳洁瑜吴李君吴跃进
Owner NANJING AGRICULTURAL UNIVERSITY
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