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Tabletting method for locust stem tip chromosome

A chromosome and stem tip technology, applied in the field of plant identification, can solve the problems of poor dispersion of cells, few literature reports, chromosome damage, etc., and achieve the effect of light dyeing color, high repeatability, uniform and clear dyeing

Inactive Publication Date: 2009-07-15
天津市林业果树研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Stem tips of tough forest trees are more difficult to dissociate, the dissociation time is short, and the cells cannot be dispersed well; the dissociation time is long, and the chromosomes will be damaged
At present, the root tip is a common material for making chromosome pellets, and there are few literature reports on the method of making chromosome pellets from the shoot tip of Robinia pseudoacacia

Method used

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  • Tabletting method for locust stem tip chromosome
  • Tabletting method for locust stem tip chromosome
  • Tabletting method for locust stem tip chromosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Chromosome Pressing of Stem Tip of Erqiao Black Locust Tissue Cultured Seedlings

[0054] In this example, the shoot tip of Robinia pseudoacacia tissue-cultured seedlings was used as a sample to prepare the shoot tip chromosome press, which specifically includes the following steps:

[0055] 1. The material of the shoot tip: the temperature of the tissue culture room is 25°C, and the shoot tip of Erqiao Robinia pseudoacacia seedlings for tissue culture is taken at 3 pm, the leaves and petioles on the stem are removed with tweezers, and 3-5 mm is cut from the shoot tip .

[0056] 2. Pretreatment of shoot tips: put the shoot tips in 0.0035M 8-hydroxyquinoline for 3 hours.

[0057] 3. Fixation of the stem tip: Take the stem tip out of the pretreatment solution with tweezers, rinse it with distilled water for 3-5 times, put the stem tip into the fixative solution and fix it for 24 hours.

[0058] 4. Dissociation of the shoot tip: Take the shoot tip out of the fixative wit...

Embodiment 2

[0064] Chromosome Pressing of Stem Tip of Erqiao Black Locust Tissue Cultured Seedlings

[0065] 1. The material of the stem tip: the temperature of the tissue culture room is 22°C, and the stem tip of Erqiao Robinia pseudoacacia seedlings for tissue culture is taken at 4 pm, the leaves and petioles on the stem are removed with tweezers, and 3-5 mm is cut from the stem tip .

[0066] 2. Pretreatment of shoot tips: put the shoot tips in 0.0025M 8-hydroxyquinoline for 5 hours.

[0067] 3. Fixation of the stem tip: Take the stem tip out of the pretreatment solution with tweezers, rinse it with distilled water for 3-5 times, put the stem tip into the fixative solution and fix it for 24 hours.

[0068] 4. Dissociation of the shoot tip: Take the shoot tip out of the fixative with tweezers, rinse it with distilled water for 3-5 times, put it in 1N HCl, and dissociate it in a 60°C water bath for 8 minutes.

[0069] 5. Slicing of the shoot tip: Take out the shoot tip from 1N HCl with...

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Abstract

The invention provides a robina stem-apex chromosome pressing method, comprising: taking material, preprocessing, fixing, dissociating, pressing, dyeing, striking, microscopic examining, characterized in that: taking the stem-apex of robina tissue culture seedling in vigorous growth stage at 3-5pm at the culture-room temperature 22-25 DEG C., wherein the cell of the stem-apex of robina tissue culture seedling is at vigorous division stage; preprocessing the stem-apex using 8-hydroxy quinoline 0.002-0.004m for 3-5 hours; pressing the stem-apex using a cross pressing method; dyeing the pressed stem-apex using carbol fuchsin; striking the material using a pencil head and uniformly dispersing the cells to the best. The robina stem-apex chromosome pressing method has features of simple operation, high pressing efficiency, saved medicine consumption, uniform chromosome dispersion, easy counting, very easy observation of metaphase chromosome, uniform and clear dyeing, especially suitable for chromosome pressing of tough wood material.

Description

technical field [0001] The invention belongs to the technical field of plant identification and relates to a method for pressing chromosomes of forest tree shoot tips. It is especially suitable for the compression of shoot tip chromosomes of forest trees such as Robinia pseudoacacia, which are tough and rich in meristem cytoplasm and intercellular pectin. Background technique [0002] Many traits of organisms are controlled by genes, and genes are mainly located on chromosomes in the nucleus. Therefore, changes in the morphological structure or quantity of chromosomes will inevitably cause variations in the traits of organisms. Distant hybridization, radiation breeding, haploid breeding, polyploid breeding, etc. are one of the important ways of crop breeding, and to identify whether their offspring are real distant hybrids, what type of variation, etc., through the genetic behavior of chromosomes The analysis is an important method that is more accurate and common. Theref...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/84G01N1/28
Inventor 黄建全张娜朱延林田淑芬刘建华苗博瑛
Owner 天津市林业果树研究所
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