Method for sheeting peanut root tip cell chromosome in mitosis metaphase

A technology of mitosis and peanut root, applied in the field of cytogenetic production, can solve the problems such as enzymatic hydrolysis time, low osmotic time difficult to grasp, great influence on chromosome structure, unsuitable control of dissociation time, etc., and achieves good operability, Save experimental time and improve production efficiency

Inactive Publication Date: 2012-07-18
HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the process of removing the cell wall by enzymatic hydrolysis and hypotonicity, the enzymatic hydrolysis time and hypotonic time are not easy to grasp, and the processing time of different plant materials may be different, resulting in more chromosome loss; The treatment time may be different for different plant materials; in addition, hydrochloric acid has a great influence on the structure of chromosomes. If the dissociation time is too short, the cells will not be pressed; if the dissociation time is too long, the chromosomes will be severely damaged, and the test results are not ideal.

Method used

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  • Method for sheeting peanut root tip cell chromosome in mitosis metaphase
  • Method for sheeting peanut root tip cell chromosome in mitosis metaphase
  • Method for sheeting peanut root tip cell chromosome in mitosis metaphase

Examples

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Effect test

Embodiment 1

[0029] Example 1: Method for producing metaphase chromosomal chromosomes in root tip cells

[0030] Taking peanut cultivars as an example, the specific method is as follows:

[0031] (1) Pretreatment: Soak peanut seeds in clean water at 25°C for 1 day. After the seeds are white, put the seeds on moist absorbent paper and keep the absorbent paper moist. Lateral roots will grow in 6-8 smallpox. Select a 10-15 mm long, thick Fresh lateral roots of 0.8-1.2 mm were soaked in a saturated aqueous solution of p-dichlorobenzene for 3 hours at room temperature;

[0032] (2) Fixation: The treated lateral roots were transferred to Carnoy's fixative solution, treated at room temperature for 12 hours, and then placed in a -20°C refrigerator for use. Wherein the composition of Carnot's fixative is: alcohol: the volume ratio of glacial acetic acid is 3:1;

[0033] (3) Acetic acid tableting: Take the fixed root tip and soak it in acetic acid with a concentration of 45% (V / V) for 30-60 seco...

Embodiment 2

[0040] Example 2: Method for producing metaphase chromosomal chromosomes in root tip cells

[0041] Taking tomato as an example, the specific method is as follows:

[0042] (1) Put the tomato seeds on the absorbent paper petri dish, pour water to soak the seeds for 1 day, pour off the water after the seeds are white, keep the absorbent paper moist, and cut the roots after 2-3 days when the seed roots grow to 10-15 mm. Soak in saturated aqueous solution of p-dichlorobenzene for 3 hours.

[0043] (2) Transfer the roots to Carnoy's fixative solution of alcohol: glacial acetic acid = 3:1, treat at room temperature for 24 hours, and put them in a -20°C refrigerator for tablet production.

[0044] (3) Acetic acid pressing: Take the fixed root tip and soak it in 45% (V / V) acetic acid for 30-60 seconds. Put a drop of 45% (V / V) acetic acid on the cover glass, adjust the root tip to the center of the cover glass, pad the corner of the cover glass with a double-sided blade, and tap t...

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Abstract

The invention relates to a method for sheeting peanut root tip cell chromosome in the mitosis metaphase, which comprises the following steps that: a peanut root tip is selected, and paradichlorobenzene is utilized for soaking pretreatment; the root tip is put into carnoy stationary liquid for room temperature treatment; acetic acid is used for soaking, the root tip of a meristematic zone is cut and put on a glass slide, the center of a cover glass is tapped to disperse the cells, a cell wall is heated by fire for dissociating and softening, and a filter paper is used for removing the excess acetic acid; the cell which is in the mitosis metaphase and has smoothly dispersed chromosome is sheeted and refrigerated, the cover glass is removed, and the glass slide is dehydrated; and stain zoning or fluorescence in-situ hybridization are carried out. The obtained sheet has leveled cells and dispersed chromosome, and more cells are in the metaphase; and the acetic acid medium is used for direct compression, the reagent has small influence to the chromosome, so the method is easy to operate and control, has simple steps and high sheeting efficiency. The method is suitable to not only peanuts but also sesame, paddy, corns, soybeans, green beans or tomatoes and other various plants, and has good operability.

Description

technical field [0001] The invention relates to a method for producing cytogenetic slices, in particular to a method for producing slices in the metaphase of chromosome mitosis of peanut root tip cells. Background technique [0002] Plant chromosomes are the material basis of cell inheritance. Variations in chromosome structure and number provide abundant genetic resources for plant breeding, and root tip cell mitotic metaphase production provides a good technical means for studying plant chromosomes. However, plant cells have solid cell walls, and it is difficult for chromosomes to spread evenly on a glass slide. In order to obtain a good chromosome preparation in the mid-mitosis stage of plant root tip cells, the researchers used hydrochloric acid to dissociate and soften the cell wall at first, and later used the method of enzymatic hydrolysis to remove the wall and low osmosis. has wide application. However, in the process of removing the cell wall by enzymatic hydroly...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
Inventor 张新友杜培易明林黄冰艳汤丰收董文召
Owner HENAN ACAD OF AGRI SCI
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