Flaking method for distantly-hybridized sugarbeet chromosome

A technology of chromosome preparation and distant hybridization, which is applied in the field of distant hybridization sugar beet chromosome preparation, can solve the problems of micro-cutting, short chromosomes, and affecting probe penetration, etc., to achieve easy operation and mastery, improve work efficiency and experimental efficiency Improved effect

Inactive Publication Date: 2019-03-19
HEILONGJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Distant hybrid sugar beet has shorter chromosomes, making it difficult to discern additional chromosomes, a problem that has long puzzled researchers
[0004] (1) The cell wall and cytoplasm of plant cells severely cover the chromosomes, making it difficult to identify and identify additional chromosomes. In addition, the low mitotic index is not easy to obtain a large number of metaphase divisions, which makes the chromosome microsegregation and in situ hybridization and additional chromosomes of distant hybridization sugar beet difficult. Chromosomal signal analysis of white flower beet brings great difficulties
(2) Conventional methods bring great difficulties to chromosome micro-segregation and micro-cloning, such as: first, it is difficult to identify additional chromosomes; second, micro-separation and micro-cutting of difficult-to-identify chromosomes cannot be performed; It is the cytoplasmic coverage of the chromosome that affects the penetration of the probe
It is even more unfavorable for in-depth research on distant hybrid materials

Method used

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  • Flaking method for distantly-hybridized sugarbeet chromosome
  • Flaking method for distantly-hybridized sugarbeet chromosome
  • Flaking method for distantly-hybridized sugarbeet chromosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] 1. Sampling: Take the new leaves of distant hybrid sugar beet at 8:00-10:00 in the morning every day;

[0053] 2. Pretreatment: pretreat sugar beet heart leaves or root tips with 8-hydroxyquinoline, and the pretreatment time is 2-4 hours;

[0054] 3. Washing: Wash the beet leaves with distilled water for 2-3 times;

[0055] 4. Dissociation: dissociate the washed beet leaves with dissociation solution for 5-10 minutes, and rinse with distilled water for 2-3 times after dissociation;

[0056] 5. Staining and pressing: stain with the improved fuchsin staining solution for 2-3 minutes, cover the stained material with a cover glass, absorb the excess staining solution with absorbent paper, and use tweezers or one end of an elastic marker pen Tap the coverslip moderately to remove air bubbles;

[0057] 6. Microscopic examination observation: Find the phase of metaphase chromosomal division, observe and count, and seal the slide with Canadian gum or nail polish.

Embodiment 2

[0059] 1. Sampling: Take the new leaves of the distant hybrid sugar beet at 8:00-10:00 in the morning every day; the distant hybrid sugar beet is the monomer addition line beet M14 or allotriploid sugar beet;

[0060] 2. Pretreatment: pretreat sugar beet heart leaves or root tips with 8-hydroxyquinoline, and the pretreatment time is 2-4 hours;

[0061] 3. Washing: Wash the beet leaves with distilled water for 2-3 times;

[0062] 4. Dissociation: dissociate the washed beet leaves with dissociation solution for 5-10 minutes, and rinse with distilled water for 2-3 times after dissociation;

[0063] 5. Staining and pressing: stain with the improved fuchsin staining solution for 2-3 minutes, cover the stained material with a cover glass, absorb the excess staining solution with absorbent paper, and use tweezers or one end of an elastic marker pen Tap the coverslip moderately to remove air bubbles;

[0064] 6. Microscopic examination observation: Find the phase of metaphase chromo...

Embodiment 3

[0066] 1. Sampling: Take the new leaves of distant hybrid sugar beet at 8:00-10:00 in the morning every day;

[0067] 2. Pretreatment: pretreat sugar beet heart leaves or root tips with 8-hydroxyquinoline for 2-4 hours; the concentration of 8-hydroxyquinoline is 0.002mol / L.

[0068] 3. Washing: Wash the beet leaves with distilled water for 2-3 times;

[0069] 4. Dissociation: dissociate the washed beet leaves with dissociation solution for 5-10 minutes, and rinse with distilled water for 2-3 times after dissociation;

[0070] 5. Staining and pressing: stain with the improved fuchsin staining solution for 2-3 minutes, cover the stained material with a cover glass, absorb the excess staining solution with absorbent paper, and use tweezers or one end of an elastic marker pen Tap the coverslip moderately to remove air bubbles;

[0071] 6. Microscopic examination observation: Find the phase of metaphase chromosomal division, observe and count, and seal the slide with Canadian gum...

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Abstract

The invention belongs to the technical field of plant flaking and discloses a flaking method for a distantly-hybridized sugarbeet chromosome. The flaking method comprises the steps: sampling, namely selecting young leaves of distantly-hybridized sugarbeet; pretreatment, namely pretreating interior leaves or root tips of sugarbeet by using 8-hydroxyquinoline for 2-4 h; washing; dissociation; dyeingand tabletting; and microscopic examination observation, namely finding a split phase of a metaphase chromosome, and carrying out observation and counting. The flaking method is beneficial to the recognition and observation of the distantly-hybridized sugarbeet chromosome; the pretreatment time is prolonged; and the condition of the additional chromosome of the distantly-hybridized sugarbeet canbe clearly observed, and the experiment efficiency can be remarkably increased. The flaking method is simple, convenient and feasible, low in cost, good in effect, easy to operate and master and capable of increasing the work efficiency, saving time and labor and realizing observation by using a common optical microscope. A monomer addition line sugarbeet M14, allotriploid sugarbeet, beta corolliflora and common cultivated sugarbeet are respectively flaked and observed by using an improved flaking method, and the flaking method is very good in effect and repeatable. The method is also suitablefor a dropping method and a smearing method.

Description

technical field [0001] The invention belongs to the technical field of plant slice production, and in particular relates to a method for producing distant hybrid sugar beet chromosome slices. Background technique [0002] At present, the commonly used prior art in the industry is as follows: observation of chromosome preparation is one of the main research methods of plant cell biology. Chromosome preparation and observation usually adopts the compression method. Because sugar beet chromosomes are very short. However, it is more difficult to prepare chromosomes of distant hybrid beet. Mainly it is difficult to identify additional chromosomes. The short chromosomes of distant hybrid sugar beets make it difficult to discern additional chromosomes, a problem that has long puzzled researchers. Can't solve this difficult problem with conventional tabletting method. Therefore, the in-depth research on distant hybrid sugar beet is also very limited. [0003] In summary, the p...

Claims

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Application Information

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IPC IPC(8): G01N1/28G01N1/30
CPCG01N1/2813G01N1/30G01N2001/302
Inventor 刘丽萍
Owner HEILONGJIANG UNIV
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