Fluorescence labeling method for cotton pollen tube microfilament framework

A technology of fluorescent labeling and pollen tube, applied in the biological field, can solve problems such as unreported, and achieve the effect of high fluorescent dyeing efficiency and good labeling effect

Inactive Publication Date: 2011-11-23
NANJING AGRICULTURAL UNIVERSITY
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  • 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
  • Fluorescence labeling method for cotton pollen tube microfilament framework

Examples

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Embodiment 1

[0021] Example 1. Fluorescence marking of pollen tube microfilament skeleton of Luxian 21 and its microscope observation

[0022] First, the method of the present invention is used to fluorescently label the pollen tube of Lumian 21 (species for production), including the following steps:

[0023] 1) Punch a hole on the glass slide. Use an electric drill to drill a hole with a diameter of 1cm in the middle of the slide glass (25.5×76.2mm, 1mm-1.2mm thick). One side of the hole is sealed with a cover glass. Adhesive For nail polish, let it stand at room temperature until the nail polish is air-dried, and the cover glass and glass slide are glued together to form a small chamber for use;

[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, dissolve in deionized water and adjust the pH to 6.2. Dilute the volume to 100ml, then add 1% low m...

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Abstract

The present 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 1to 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 the pollen tube microfilament skeleton 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. Its germination and elongation and other physiological activities directly affect the smooth completion of the fertilization process. The growth of the pollen tube is different from other plant cells. The polar apex grows and depends on the normal aggregation of microfilaments in the growing pollen tube. Therefore, using the pollen tube system to study microfilament skeletons and their functions in organisms is currently one of the most active research fields and is also the focus of research in the field of plant reproductive biology. Recent studies have shown that microfilaments are an important part of the cytoskeleton. They perform m...

Claims

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Application Information

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Patent Type & Authority Patents(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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