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Method and dissociation liquid for identifying ploidy of kiwi fruit by flow cytometry and preparation method of dissociation liquid

A technology of flow cytometer and dissociation solution, which is applied in the field of dissociation solution and preparation, can solve the problems of not identifying the dissociation solution, etc., achieve the effects of simple drug preparation, increase cell permeability, and avoid tube blockage

Pending Publication Date: 2020-02-21
四川省自然资源科学研究院
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to overcome the deficiencies of the prior art, to provide a method for identifying the ploidy of kiwifruit using flow cytometry, a dissociation solution and a preparation method, which solves the problem that there is no dissociation solution for the ploidy identification of kiwifruit in the prior art The problem

Method used

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  • Method and dissociation liquid for identifying ploidy of kiwi fruit by flow cytometry and preparation method of dissociation liquid
  • Method and dissociation liquid for identifying ploidy of kiwi fruit by flow cytometry and preparation method of dissociation liquid
  • Method and dissociation liquid for identifying ploidy of kiwi fruit by flow cytometry and preparation method of dissociation liquid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A dissociation solution suitable for identifying kiwifruit ploidy by flow cytometry, including components and corresponding concentrations as shown in Table 1:

[0034] Table 1

[0035]

[0036]

[0037] The above-mentioned preparation method that is applicable to the dissociation liquid of flow cytometry identification kiwifruit ploidy comprises: taking by weighing 3.725g potassium chloride (KCl), 2.46g magnesium sulfate heptahydrate (MgSO 4 ·7H 2 O), 1.19g hydroxyethylpiperazine ethylsulfuric acid (HEPES), 10.00g polyvinylpyrrolidone (PVP-30) and 2.5mL polyethylene glycol octylphenyl ether (Triton X-100), a small amount of several times Stir to dissolve, set the volume to 1000mL, adjust the pH to 7.5 with NaOH solution, and store at 4°C.

[0038] Materials and Reagents Required for Identifying Ploidy of Kiwifruit by Flow Cytometry

[0039] Pipette tips, blades (Blue Gillette), 40 μm cell mesh (Suohua Cat. No.: 251100), dissociation solution, PI staining solut...

Embodiment 2

[0049] A dissociation solution suitable for flow cytometry identification of kiwifruit ploidy, including components and corresponding concentrations as shown in Table 2:

[0050] Form 2

[0051]

[0052] The preparation method of the above-mentioned dissociation solution suitable for flow cytometry identification of kiwifruit ploidy is the same as in Example 1, and the volume is also adjusted to 1000mL, the pH is adjusted to 7.3 with NaOH solution, and stored at 4°C.

[0053] Materials and Reagents Required for Identifying Ploidy of Kiwifruit by Flow Cytometry

[0054] Pipette tips, blades (Blue Gillette), 40 μm cell mesh (Suohua Cat. No.: 251100), dissociation solution, PI staining solution, filter paper, disposable gloves, Petri dish, 1.5mL centrifuge tube, ice box

[0055] Apparatus and equipment required for identification of kiwifruit ploidy by flow cytometry

[0056] Flow cytometer (BD), pipette gun, ice maker, cryogenic centrifuge

[0057] The operation steps requ...

Embodiment 3

[0064] A kind of dissociation solution suitable for identifying kiwifruit ploidy by flow cytometry, including components and corresponding concentrations as shown in Table 3:

[0065] Form 3

[0066]

[0067] The preparation method of the above-mentioned dissociation solution suitable for flow cytometry identification of kiwifruit ploidy is the same as in Example 1, and the volume is also adjusted to 1000mL, the pH is adjusted to 7.2 with NaOH solution, and stored at 4°C.

[0068] Materials and Reagents Required for Identifying Ploidy of Kiwifruit by Flow Cytometry

[0069] Pipette tips, blades (Blue Gillette), 40 μm cell mesh (Suohua Cat. No.: 251100), dissociation solution, PI staining solution, filter paper, disposable gloves, Petri dish, 1.5mL centrifuge tube, ice box

[0070] Apparatus and equipment required for identification of kiwifruit ploidy by flow cytometry

[0071] Flow cytometer (BD), pipette gun, ice maker, cryogenic centrifuge

[0072] The operation steps...

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Abstract

The invention relates to the technical field of kiwi fruit breeding, in particular to a dissociation liquid for identifying ploidy of kiwi fruit by a flow cytometry. The molar concentration of inorganic salt in the dissociation liquid is 0.8-1.2 mol / L, the molar concentration of a buffer agent in the dissociation liquid is 3-10 mmol / L, the mass percent of polyvinylpyrrolidone in the dissociation liquid is 0.8-1.2%, the mass percent of p-octyl polyethylene glycol phenyl ether in the dissociation liquid is 0.2-0.3%, and pH of the dissociation liquid is 7.0-8.0. According to the preparation method, all substances are stored at a low temperature after melting in a mixed way. The method for identifying the ploidy of the kiwi fruit by the flow cytometry comprises the steps of dying after dissociation and centrifugation of kiwi fruit cells in the dissociation liquid for many times, and detecting by a machine. By the method, the problem of no dissociation liquid for identifying the ploidy of the kiwi fruit in the prior art is solved, the provided dissociation liquid is suitable for cell extraction of a kiwi fruit sample, a drug is simple to prepare, and the method is suitable for mass detection.

Description

technical field [0001] The invention relates to the technical field of kiwifruit breeding, in particular to a method for identifying kiwifruit ploidy by using a flow cytometer, a dissociation solution and a preparation method. Background technique [0002] The DNA content and ploidy level of plant nuclei are crucial to the study of biological diversity, and can be widely used in many research fields of basic and applied botany. The basis of these studies is the C value, the total amount of DNA contained in the haploid nucleus in eukaryotic cells. Based on flow cytometry (FCM), flow cytometry can quickly and accurately determine the DNA content of the G1 phase nucleus of plant samples, reflecting cell ploidy. Flow cytometry is widely used in many research fields such as plant genetics and breeding, plant taxonomy, stress botany, and plant pathology. [0003] Kiwifruit (kiwifruit) is a perennial vine belonging to the genus Actinidia Lindl of the family Actinidiaceae. There ...

Claims

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Application Information

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IPC IPC(8): G01N1/28G01N15/14G01N21/64
CPCG01N1/28G01N15/1434G01N21/6428G01N2021/6439
Inventor 张茜王丽华李峤虹庄启国李明章郑晓琴何利钦
Owner 四川省自然资源科学研究院
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