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Paddy rice root tip chromosome slide-making method

A chromosome production and rice root technology, applied in the biological field, can solve the problems of difficult microscopic examination, difficult to count, complicated production procedures, etc., and achieve the effects of easy observation, convenient operation, and easy microscopic examination.

Inactive Publication Date: 2008-09-17
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the limitation of small chromosomes in rice, the preparation of rice chromosomes is more difficult than other plants.
[0003] Found through document retrieval to prior art, Xie Guosheng etc. compared in " the comparative study of rice chromosome production technology " article published in " China Agricultural Science Bulletin " 1998, the 14th volume, the 3rd phase, 27-28 page Rice root tip chromosome compression method and smear method are two kinds of production techniques: the compression method is convenient for dissociation and processing, easy to microscopic examination, simple production but the background is thicker, and the chromosomes are difficult to separate and count; the background of the smear method is very light , the chromosomes are easy to separate, but there are few cells in the field of view, the microscopic examination is difficult, and the production procedure is cumbersome
Both methods have certain disadvantages

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Nine plants were obtained from the anther culture of the rice variety "Taipei 309". At 20:00, white new roots with a length of 2 cm were taken from rice plants; pretreated with ice water for 12 hours, mixed with 6% enzyme solution, and enzymolyzed at 30°C for 2.5 hours; washed with distilled water for 15 seconds, blotted dry with absorbent paper, and fixed by immersing in modified CarnoyI solution (70% alcohol: glacial acetic acid = 3:1, V / V) for 30 min; wash with distilled water for 5 s, and soften with 45% acetic acid for 10 min; The root tip part before sink marks, stained with modified phenol fuchsin (containing 0.246g / L fuchsin, 3.69g / L phenol, 18g / L sorbitol, 0.978% acetic acid, 0.896% formalin) for 1min, after pressing Observe under a light microscope.

[0031] Microscopic examination effect: under the condition of 1000 times magnification (10 times eyepiece × 100 times objective lens), more than 85% of the apical mitosis cells have light background, well disper...

Embodiment 2

[0033] Eighteen plants were obtained from the anther culture of the rice variety "Nipponbare". At 11:00, take white new roots with a length of 2 cm on the rice plants; pretreat with ice water for 24 hours, mix with 6% enzyme solution, and enzymatically hydrolyze at 29°C for 3 hours; wash with distilled water for 15 seconds, blot dry with absorbent paper, and immerse in the modified CarnoyI fixative solution (70% alcohol: glacial acetic acid = 3:1, V / V) fixed for 45 min; washed with distilled water for 5 s, softened with 45% acetic acid for 10 min; put the treated rice root tip on a glass slide, and cut out the constriction that appeared after enzymatic hydrolysis The root tip part before the scar was stained with modified phenol fuchsin (containing 0.246g / L magenta, 3.69g / L phenol, 18g / L sorbitol, 0.978% acetic acid, 0.896% formalin) for 1min, and pressed into pieces; Eupara glue was mounted around the coverslip, and observed under an optical microscope 1 day later.

[0034] ...

Embodiment 3

[0036] One plant obtained from the anther culture of the rice variety "Zhonghua 11". At 21:00, white new roots with a length of 1.5 cm were taken from rice plants; pretreated with ice water for 18 hours, mixed with 6% enzyme solution, and enzymatically hydrolyzed at 29°C for 3 hours; washed with distilled water for 15 seconds, blotted dry with absorbent paper, and fixed by immersing in modified CarnoyI solution (70% alcohol: glacial acetic acid = 3:1, V / V) for 1 day; washed with distilled water for 5 s, and softened with 45% acetic acid for 10 min; put the treated rice root tip on a glass slide, cut out the The root tip before the constriction mark was stained with modified phenol fuchsin (containing 0.246g / L fuchsin, 3.69g / L phenol, 18g / L sorbitol, 0.978% acetic acid, 0.896% formalin) for 1min, and pressed into slices ; Use Eupara glue to mount around the coverslip, and observe under an optical microscope after 1 week.

[0037] Microscopic examination effect: under the condi...

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Abstract

A sheet forming method of paddy root tip, belongs to biotechnology domain. The method comprises: pretreating by ice water; dissociating by cellulase and pectinase; dyeing using phenol fuchsin; fixing using CarnoyI fixation fluid; sheet forming by pressed disk method. The sheet forming method uses cellulase and pectinase to dissociate and forms the sheet by pressed disk method, with virtues of conveniently operating and easily testing under microscope compared with the traditional smear method and with virtues of light background and dispersed chromosome compared with the traditional pressed disk method. The invention solves the defects of complicated operation of forming the sheet of the paddy root tip and difficulty of observing and counting the chromosome.

Description

technical field [0001] The invention relates to a method in the field of biotechnology, in particular to a method for preparing rice root tip chromosomes. Background technique [0002] As an important food crop and model plant, the research on rice genetics and breeding is developing rapidly. However, due to the limitations of the small chromosomes of rice, the production of rice chromosomes is more difficult than that of other plants. [0003] Found through document retrieval to prior art, Xie Guosheng etc. compared in " the comparative study of rice chromosome production technology " article published in " China Agricultural Science Bulletin " 1998, the 14th volume, the 3rd phase, 27-28 page Rice root tip chromosome compression method and smear method are two kinds of production techniques: the compression method is convenient for dissociation and processing, easy to microscopic examination, simple production but the background is thicker, and the chromosomes are difficul...

Claims

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Application Information

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IPC IPC(8): G01N1/30
Inventor 蒋苏蔡润潘俊松何欢乐
Owner SHANGHAI JIAO TONG UNIV
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