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Wheat root tip chromosome production method

A technology of chromosome preparation and root tip, which is applied in the field of plant genetics, can solve the problems of easy loss of glass slide division, weak chromosome attachment, chromosome breakage, etc., and achieve the effect of clear division phase, not easy to lose, and weak background signal

Inactive Publication Date: 2016-09-21
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These two methods have the following problems in the operation process: when the cover glass is removed by the pressing method, the cleavage phase on the slide is easy to lose; the pressing method sometimes causes chromosome breakage; the background is relatively dark
Chromosomes are not firmly attached to the glass slide by the dropping method, and chromosomes are likely to be lost during subsequent experiments; sometimes the background is dark; sometimes special equipment is required to pretreat the material, etc., which cannot Meet the needs of ordinary laboratories for chromosome microscopic examination

Method used

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  • Wheat root tip chromosome production method
  • Wheat root tip chromosome production method
  • Wheat root tip chromosome production method

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Example 1 Chromosome preparation of common wheat Chinese spring (2n=6x=42)

[0028] Seed rooting: put 30 common wheat varieties of Chinese spring seeds in a petri dish with filter paper, add enough distilled water to fully soak the seeds in distilled water, and place them for 20 hours at room temperature and under shading conditions; after the seeds are white, After absorbing excess water, move to a 4°C refrigerator and shading treatment for 24 hours; then place the petri dish in a 22°C incubator to grow in the dark.

[0029] Pretreatment: When the root of the wheat seed grows to 0.5-1.5cm, cut off all the young root, put it into a 1.5ml eppendorf tube with 1ml concentration of 2mM 8-hydroxyquinoline solution, and keep it under dark conditions at 22°C 2.5 hours for preprocessing (in actual operation, the preprocessing time can be within 2-4 hours);

[0030] Fixing: Take out the pretreated root tip, absorb the excess 8-hydroxyquinoline solution with absorbent paper, was...

Embodiment 2

[0034] Example 2 Chromosome preparation and fluorescence in situ hybridization of durum wheat (2n=4x=28) and wheat tufts (2n=2x=14)

[0035] Rooting of seeds: Place the seeds of durum wheat variety Zhongyin 1286 and A. villosa (Chen Peidu et al., Cytogenetics of hybrid wheat and A. villosa, Journal of Nanjing Agricultural University, 1982, 4:1-15) Add enough distilled water to a petri dish lined with filter paper to fully soak the seeds in distilled water, and place it for 20 hours at room temperature and under light-shading conditions; after the seeds are white, absorb excess water, move to a 4°C refrigerator, and shading treatment for 24 hours. hours; then put the petri dish in a 22°C incubator to grow in the dark.

[0036] Pretreatment: put the clipped fresh radicle into a 1.5mleppendorf tube containing 1ml of 2mM 8-hydroxyquinoline solution, and pretreat for 3h at 22°C in the dark;

[0037] Fixing: Take out the pretreated root tip, absorb the excess 8-hydroxyquinoline solut...

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Abstract

The invention discloses a wheat root tip chromosome production method. The method includes the specific steps that after seeds germinate, rootlets are taken, preprocessed with 8-hydroxyquinoline and then subjected to fixation with Kano fixing fluid, low-osmolarity preprocessing with distilled water, enzymolysis with cellulase and pectinase and low-osmolarity postprocessing with distilled water, the processed root tip growth points are subjected to chromosome production through a smearing method, and finally microscopic examination is conducted under a phase contrast microscope. The chromosome production method is convenient and easy to operate, chromosome dispersity is good, the background is relatively shallow, counting is easy, and a splitting cell is not prone to losing.

Description

technical field [0001] The invention relates to the field of plant genetics, in particular to a method for preparing wheat root tip chromosomes. Background technique [0002] Wheat is an important food crop in the world. There are more than 20 species in the Triticum genus. Common wheat belongs to the Gramineae Triticum family, the Triticum subfamily, and the Triticum genus. Its cultivated area accounts for about 90% of the total area of ​​wheat. The genera in the wheat cluster and the species in the Triticum genus except common wheat are generally called relative plants of common wheat (Wu Zhaosu, Wheat Breeding, Agricultural Press, 1990). The close relatives of common wheat contain a variety of trait genes that are not available in common wheat itself and can be used for variety improvement. Through distant hybridization and chromosome engineering technology, these genes can be transferred to common wheat In order to enrich the genetic basis of common wheat. Chromosome r...

Claims

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Application Information

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IPC IPC(8): G01N1/28
CPCG01N1/28
Inventor 杨学明张鹏周淼平姚金保马鸿翔张瑜余桂红
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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