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A kind of Chromosome preparation method of the root tip of a plant of the genus Lyme

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: 2016-06-08
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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  • A kind of Chromosome preparation method of the root tip of a plant of the genus Lyme

Examples

Experimental program
Comparison scheme
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] ⑵Under the condition of 25°C, place the centrifuge tube with the apex in step ⑴ in a high-pressure container device, and inject 0.4MPaN 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 according to the ratio of 3mL: 1mL.

[0025] (4) Rinse the fixed new roots with distilled water...

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 0.8MPaN 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 volume c...

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] ⑵Under the condition of 26°C, place the centrifuge tube with the apex in step ⑴ in a high-pressure container device, and inject 0.6MPaN 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 ...

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Abstract

The present invention relates to a kind of method for producing root tip chromosomes of plants of the genus Lysymus genus, the method comprising the following steps: (1) taking materials: 5 to 7 days after the seeds of Lysymus germinate at a constant temperature, take the white new roots of vigorously growing potted plant Lysymus spp. Put it into a centrifuge tube; (2) place the centrifuge tube with the root tip in a high-pressure container device, and pretreat it with N2O to obtain a new root after pretreatment; (3) dry the new root after pretreatment, and use Carnot’s fixed with liquid to obtain new roots after fixation; (4) the new roots after the fixation were rinsed and dried and then dissociated with glacial acetic acid to obtain new roots after the dissociation; (5) the moisture in the new roots after the dissociation was blotted and placed On the glass slide, make a pressed piece; (6) place the pressed piece under an optical microscope to observe, select the cells with dispersed and clear chromosomes for photography, so as to perform fluorescence in situ hybridization, and finally mark the position of the cover glass with a glass knife, Store the prepared slices in a -80°C refrigerator. The invention 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 Patents(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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