Method for preparing paeonia plant root tip chromosome slice

A technology for chromosome preparation and Paeonia lactiflora is applied in the field of chromosome preparation at the root tip of Paeonia lactiflora, can solve the problems of large influence of preparation results, high requirements for preparation conditions, influence on karyotype analysis results, and the like, and achieves low cost , Expand the range of materials, and increase the effect of the concentration of hydrochloric acid

Inactive Publication Date: 2020-05-19
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the scope of this method is relatively narrow, and only the root tip chromosomes of peony are prepared. The dissociation steps in this patent are similar to the normal dissociation steps, which need to be carried out at 55-65°C, and the requirements for the production conditions are relatively high. The process takes a long time, and in the process of conventional root tip chromosome preparation, the cell wall and cytoplasm have a great influence on the preparation results, and under high temperature and acidic conditions, the number of good cleavage phases is small and the chromosomes are easy to break and decompose, which affects the nucleus. type analysis results

Method used

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  • Method for preparing paeonia plant root tip chromosome slice
  • Method for preparing paeonia plant root tip chromosome slice
  • Method for preparing paeonia plant root tip chromosome slice

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] (1) Material collection: 1-1.5 cm of the white root tips of new roots germinated in autumn from the vigorously growing peony ‘Xishi Fen’ seedlings in the field were excavated from 9 to 11 in the morning in late September 2019 (by figure 1 shown), put it into a container filled with clean water and bring it back to the laboratory, and the time from the next step of processing should not exceed 2 hours.

[0035] (2) Pretreatment: Clean the removed root tips, put them into a saturated aqueous solution of p-dichlorobenzene, and pretreat them for 3.5 hours under refrigeration at 0-4°C.

[0036] (3) Fixation: Rinse the pretreated root tips three times with distilled water, blot dry, and place them in newly prepared Carnot’s fixative solution (absolute ethanol: glacial acetic acid = 3:1) at 0-4°C for refrigeration. Fixed for 22-24 hours.

[0037] (4) Dissociation: Rinse the fixed root tip three times with distilled water, blot dry the water, put it into a dissociation solutio...

Embodiment 2

[0045](1) Materials collection: 1-1.5 cm of white root tips of new roots germinated in autumn from the vigorously growing peony 'Fengdan' seedlings in the field were excavated from 9 to 11 in the morning in late September 2019 (by image 3 shown), put it into a container filled with clean water and bring it back to the laboratory, and the time from the next step of processing should not exceed 2 hours.

[0046] (2) Pretreatment: Clean the removed root tips, put them into a saturated aqueous solution of p-dichlorobenzene, and pretreat them for 3.5 hours under refrigeration at 0-4°C.

[0047] (3) Fixation: Rinse the pretreated root tips three times with distilled water, blot dry, and place them in newly prepared Carnot’s fixative solution (absolute ethanol: glacial acetic acid = 3:1) at 0-4°C for refrigeration. Fixed for 22-24 hours.

[0048] (4) Dissociation: Rinse the fixed root tip three times with distilled water, blot dry the water, put it into a dissociation solution with...

Embodiment 3

[0056] (1) Material collection: From 9 to 11 in the morning in late September 2019, dig out 1 to 1.5 cm of the white root tip of the new root of the peony 'Kao' seedling that grows vigorously in the field in autumn, put it in a container filled with clean water and bring it back In the laboratory, the time to the next step of processing should not exceed 2 hours.

[0057] (2) Pretreatment: Clean the removed root tips, put them into a saturated aqueous solution of p-dichlorobenzene, and pretreat them for 3.5 hours under refrigeration at 0-4°C.

[0058] (3) Fixation: Rinse the pretreated root tips three times with distilled water, blot dry, and place them in newly prepared Carnot’s fixative solution (absolute ethanol: glacial acetic acid = 3:1) at 0-4°C for refrigeration. Fixed for 22-24 hours.

[0059] (4) Dissociation: Rinse the fixed root tip three times with distilled water, blot dry the water, put it into a dissociation solution with a volume ratio of absolute ethanol and ...

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Abstract

The invention relates to a chromosome flaking method of a plant and concretely relates to a method for preparing a paeonia plant root tip chromosome slice. Compared with the prior art, the method is characterized in that the material taking range of a root tip is expanded to plants in a herbaceous peony group and in a Chinese peony group, in the dissociation step, a fixed material is put into a dissociation liquid with the volume ratio of absolute ethyl alcohol to 37% hydrochloric acid being 3: 1 to be dissociated at the room temperature; and a low-permeability step in distilled water is alsoadded. Herbaceous peony and Chinese peony seedlings in a field are taken as raw materials, a special dissociation step is adopted, on one hand, the conditions required by the experiment is reduced, the experiment is more convenient; on the other hand, the concentration of hydrochloric acid is properly increased such that the influence of cell walls and cytoplasm on the flaking result is reduced, and meanwhile, the breakage and decomposition of chromosomes under a high-temperature acidic condition are also prevented; after dissociation, hyposmosis enables the chromosomes to be well dispersed, the counting is convenient, and the background is clean.

Description

technical field [0001] The invention relates to a method for preparing chromosomes of plants, in particular to a method for preparing chromosomes of root tips of Paeoniae plants. Background technique [0002] The genus Paeoniae is a shrub or herb of the Paeoniaceae plant, which is divided into three groups: the peony group, the Paeoniae peony group and the North American Paeoniae group, of which there are only two North American Paeoniae groups distributed in western North America; the Paeoniae group is the largest group in the genus, It is a herbaceous type, distributed in Eurasia, and the peony group is a woody type, endemic to my country. Most of the plants in this genus have bright flowers, among which peony and peony are dubbed "King of Flowers" and "Flower Phase" respectively. Because of its rich colors and charming flower shapes, it is widely used in gardens and gardens. [0003] Chromosomal karyotype analysis is an important indicator of cell taxonomy, and is valuab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/30
CPCG01N1/2813G01N1/30
Inventor 孙晓梅辛如洁杨宏光李雪婷宋莉莉刘雪婷
Owner SHENYANG AGRI UNIV
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