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Chromosome flaking method of leymus hochst plant root tips

A technology of chromosome production and plant root, which is applied in the field of preparation of plant chromosome metaphase specimens, can solve the problems of experimenter injury, unstable effect, long experiment time, etc., and achieve safe reagents, stable processing effect, and little physical damage Effect

Inactive Publication Date: 2014-08-27
CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Colchicine, p-dichlorobenzene, 8-hydroxyquinoline, α-bromonaphthalene and other drugs have the effect of preventing or destroying the formation of the spindle, and are highly toxic and easy to cause harm to experimenters
The low temperature method takes a long time for the experiment, and it takes 24 to 64 hours for one treatment, and the effect is unstable

Method used

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  • Chromosome flaking method of leymus hochst plant root tips

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Example 1 A method for preparing chromosomes from the root tip of a plant of the genus Lysia genus, comprising the following steps:

[0021] ⑴Materials: Lysium pratense ( L. secalinus ) 5 days after the seeds germinated at a constant temperature of 25°C, take 1.5 cm white new roots of vigorously growing potted Laymus chinensis, and put them into an open 1.5 mL centrifuge tube with water attached to the inner wall.

[0022] (2) At 25°C, place the centrifuge tube with the apex in step (1) in a high-pressure container device, and inject 0.4MPa N 2 O is pretreated, and after 3 hours, the new root after pretreatment is obtained.

[0023] (3) Dry the pretreated new roots with filter paper, fix them with Carnot's solution for 20 hours, and obtain the fixed new roots.

[0024] Among them: Carnot's solution is made by mixing absolute ethanol and glacial acetic acid at a ratio of 3 mL:1 mL.

[0025] (4) Rinse the fixed new roots with distilled water for 3 times, absorb t...

Embodiment 2

[0028] Example 2 A method for preparing chromosomes from the root tip of a plant of the genus Lysia genus, comprising the following steps:

[0029] ⑴Materials: Lysium pratense ( L. secalinus ) 7 days after the seeds germinated at a constant temperature of 25°C, take 2 cm of new white roots of vigorously growing potted Laymus chinensis, and put them into an open 1.5mL centrifuge tube with water attached to the inner wall.

[0030] (2) At 27°C, place the centrifuge tube with the apex in step (1) in a high-pressure container device, and inject 0.8MPa N 2 O is pretreated, and after 3.5h, the new root after pretreatment is obtained.

[0031] (3) Dry the pretreated new roots with filter paper, fix them with Carnot's solution for 24 hours, and obtain the fixed new roots.

[0032] Among them: Carnot's solution is the same as Example 1 .

[0033] (4) Rinse the fixed new roots with distilled water for 3 times, absorb the water and dissociate in glacial acetic acid with a volu...

Embodiment 3

[0036] Example 3 A method for preparing chromosomes from the root tip of a plant of the genus Lysia genus, comprising the following steps:

[0037] ⑴Materials: Lysium pratense ( L. secalinus ) 6 days after the seeds germinated at a constant temperature of 25°C, take 1.8cm white new roots of vigorously growing potted Laymus chinensis, and put them into an open 1.5mL centrifuge tube with water attached to the inner wall.

[0038] (2) At 26°C, place the centrifuge tube with the apex in step (1) in a high-pressure container device, and inject 0.6MPa N 2 O for pretreatment, after 3.2h, the new roots after pretreatment were obtained.

[0039] (3) Dry the pretreated new roots with filter paper, fix them with Carnot's solution for 22 hours, and obtain the fixed new roots.

[0040] Among them: Carnot's solution is the same as Example 1 .

[0041] (4) Rinse the fixed new roots with distilled water for 3 times, absorb the water and dissociate in glacial acetic acid with a volu...

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Abstract

The invention relates to a chromosome flaking method of leymus hochst plant root tips. The method comprises the following steps that (1) in a step of preparing materials, after leymus hochst seeds are germinated for 5-7 days at constant temperature, white new roots of the potted leymussecalimus which grow vigorously are put into a centrifugal tube; (2) the centrifugal tube with root tips are placed into a high pressure container device, and N2O are introduced into the centrifugal tube for pretreatment to obtain pretreated new roots; (3) the pretreated new roots are dried and fixed by using Carnoy's fluid to obtain fixed new roots; (4) the fixed new roots are rinsed and dissociated by using glacial acetic acid after being dried to obtain dissociated new roots; (5) the dissociated new roots are dried and placed on a glass slide for manufacturing tablettings; (6) the tablettings are placed under an optical microscope for observing, dispersed and distinct cells of the chromosome are selected for taking photographs so as to perform fluorescence in situ hybridization, and finally, the positions of cover glasses are marked by using a glass knife, and the ready-made glasses are put into a refrigerator at minus 80 DEG C for storing. The method is efficient and safe.

Description

technical field [0001] The invention relates to a method for preparing a plant chromosome metaphase specimen in the field of cytogenetics, in particular to a method for preparing a root tip chromosome of a plant belonging to the genus Lyymus. Background technique [0002] Chromosomes are the genetic material in the nucleus, and chromosomes of different species have relatively stable and specific morphological structures. Chromosome preparation is the basic method of cytogenetics research, and it is an indispensable and important means to study biological genetic variation, species evolution, classification, distant hybridization and the relationship between chromosome structure, shape and function. The basic process of plant chromosome preparation includes sampling, pretreatment, fixation, dissociation, staining, preparation and microscopic examination. Chromosome preparation takes the metaphase chromosomes of somatic cell division as the research object, but the duration o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
Inventor 李媛蔡振媛张毓喻凤林丽刘瑞娟
Owner CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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