Preparation method of cell chromosome metaphase specimen of psathyrostachys nevski plant

A technology for preparing plant cells and specimens, which is applied in the field of specimen preparation for metaphase mitotic phase of plants of the genus Rhizoma genus small damage effect

Inactive Publication Date: 2014-08-20
CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above-mentioned chemical drugs can prevent or destroy the formation of the spindle body, and are highly toxic and harmful to the body of the experimenter.
The low temperature method takes a long time for pretreatment, and it takes 1 to 3 days for one pretreatment, and the effect is not ideal.

Method used

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  • Preparation method of cell chromosome metaphase specimen of psathyrostachys nevski plant
  • Preparation method of cell chromosome metaphase specimen of psathyrostachys nevski plant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 A method for preparing a chromosomal metaphase sample of a plant of the genus Neotrichum, comprising the following steps:

[0022] ⑴Materials: At 9:00 am, take the vigorously growing potted new wheatgrass in the United States ( P. juncea ), and put it into an open 1.5mL centrifuge tube with water attached to the inner wall.

[0023] (2) Under the condition of 20°C, place the centrifuge tube with the apex in step (1) in the high-pressure container device, and first pass 0.4MPa N 2 O treatment for 1.5h, and then at the same temperature, N 2 O to make the partial pressure reach 0.75MPa, continue to treat for 2h50min, and obtain the new root after pretreatment.

[0024] (3) Dry the pretreated new roots with filter paper, fix them with Carnot's solution for 20 hours, and obtain the fixed new roots.

[0025] Among them: Carnot's solution is made by mixing absolute ethanol and glacial acetic acid at a ratio of 3 mL:1 mL.

[0026] (4) Rinse the fixed new ro...

Embodiment 2

[0029] Example 2 A method for preparing a chromosomal metaphase sample of a plant of the genus Neotrichum, comprising the following steps:

[0030] ⑴Materials: At 3:00 in the afternoon, the vigorously growing potted new wheatgrass in Kazakhstan ( P. juncea ), and put it into an open 2.0mL centrifuge tube with water attached to the inner wall.

[0031] (2) Under the condition of 25°C, place the centrifuge tube with the apex in step (1) in the high-pressure container device, and first pass 0.4MPa N 2 O treatment for 1.5h, and then at the same temperature, N 2 O to make the partial pressure reach 0.8MPa, continue to treat for 2.5 hours, and obtain the new root after pretreatment.

[0032] (3) Dry the pretreated new roots with filter paper, fix them with Carnot's solution for 23 hours, and obtain the fixed new roots.

[0033] Among them: Carnot's solution is the same as Example 1 .

[0034] (4) Rinse the fixed new roots with distilled water for 3 times, absorb the wat...

Embodiment 3

[0037] Example 3 A method for preparing a chromosomal metaphase sample of a plant of the genus Neotrichum, comprising the following steps:

[0038] ⑴Materials: At 3:00 p.m., vigorously growing potted new wheatgrass in the United States ( P. juncea ), and put it into an open 2.0mL centrifuge tube with water attached to the inner wall.

[0039] (2) At 26°C, place the centrifuge tube with the apex in step (1) in a high-pressure container device, and first inject 0.4MPa N 2 O treatment for 1.5h, and then at the same temperature, N 2 O to make the partial pressure reach 0.8MPa, continue to treat for 2.5 hours, and obtain the new root after pretreatment.

[0040] (3) Dry the pretreated new roots with filter paper, fix them with Carnot's solution for 23 hours, and obtain the fixed new roots.

[0041] Among them: Carnot's solution is the same as Example 1.

[0042] (4) Rinse the fixed new roots with distilled water for 3 times, absorb the water and dissociate in glacial a...

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Abstract

The invention relates to a preparation method of a cell chromosome metaphase specimen of a psathyrostachys nevski plant, which comprises the following steps: (1), obtaining materials: obtaining new roots of eugonic potted psathyrostachys juncea at any time in one day, and putting the new roots in a centrifuge tube; (2), putting the centrifuge tube filled with root tips in a high-pressure container, pre-processing by introducing N2O, and obtaining pre-processed new roots; (3), completely absorbing moisture of the pre-processed new roots, fixing by using a carnoys fixative, and obtaining fixed new roots; (4), after rinsing and completing absorbing the fixed new roots, dissociating by using glacial acetic acid, and obtaining dissociated new roots; (5), after completely absorbing moisture in the dissociated new roots, putting on a glass slide, and manufacturing a compression tablet; and (6), observing by putting the compression tablet below an optical microscope, selecting cells with dispersed and clear chromosomes to photograph, conveniently carrying out fluorescence in-situ hybridization, finally, labelling the position of a cover glass by using a glass cutter, putting the manufactured tablet in a refrigerator at minus 80 DEG C, and preserving. The preparation method of the cell chromosome metaphase specimen of the psathyrostachys nevski plant disclosed by the invention is high-efficiency and steady.

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 plant cell chromosome metaphase specimen of the genus Neotrichum. Background technique [0002] 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. [0003] Chromosome specimen preparation, that is, chromosome preparation, takes the metaphase chromosomes of somatic cell division as the research object, but the duration of metaphase is very short, generally only 10-30 minutes, so that the frequency of metaphase chromosome cells is not high. In order to overcome the above-mentioned contradictions, chemical or physical methods are commonly used to pretreat the material. Whe...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C12Q1/68
CPCG01N1/30
Inventor蔡振媛李媛张毓喻凤刘瑞娟林丽
OwnerCHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST