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Flaking method of petunia chromosome

A petunia and chromosome technology, applied in the field of petunia chromosome production, can solve the problems of incomplete elimination of cell walls, inability to disperse cells, short processing time, etc.

Inactive Publication Date: 2014-12-17
JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Plant cells rely on the cell wall and plasmodesma to connect. In order to obtain a well-dispersed mitotic metaphase, hydrochloric acid dissociation is often used. Different plant materials have different hydrochloric acid dissociation time. Short treatment time does not completely eliminate the cell wall, resulting in poor cell dispersion. On, the dissociation time is long, the chromosome structure is seriously damaged, and the staining effect is poor

Method used

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  • Flaking method of petunia chromosome
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  • Flaking method of petunia chromosome

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Example 1: Chromosome preparation using diploid petunia pistil stigma somatic cells, wherein diploid petunia 'dream' seeds were purchased from Zhejiang Hongyue Flower Co., Ltd.

[0026] The preparation method of magenta staining solution with a mass concentration of 1% is as follows: first add 45ml of acetic acid, add 55ml of water and mix well to obtain 100ml of acetic acid solution with a volume concentration of 45%, then slowly pour 1 gram of magenta powder into the above-mentioned acetic acid solution, and stir while cooking , in order to increase the dyeing effect, add 1-2 drops of 4% (mass concentration) iron alum solution after boiling and cooling, filter and use.

[0027] Carnot's fixative preparation method is: 300ml of absolute ethanol and 100ml of glacial acetic acid are evenly mixed.

[0028] The diploid petunia pistil stigma body cell carries out the method for chromosome preparation comprising the following steps:

[0029] (1) From 8:00 to 10:00 in the mo...

Embodiment 2

[0034] Example 2: Chromosome preparation using tetraploid petunia pistil stigma cells

[0035] Among them, the seeds of tetraploid petunia 'Hongxia' came from Jiangsu Agriculture and Forestry Vocational and Technical College.

[0036] The preparation method of magenta staining solution with a mass concentration of 1% is as follows: first add 45ml of acetic acid, add 55ml of water and mix well to obtain 100ml of acetic acid solution with a volume concentration of 45%, then slowly pour 1 gram of magenta powder into the above-mentioned acetic acid solution, and stir while cooking , in order to increase the dyeing effect, add 1-2 drops of 4% (mass concentration) iron alum solution after boiling and cooling, filter and use.

[0037] Carnot's fixative preparation method is: 300ml of absolute ethanol and 100ml of glacial acetic acid are evenly mixed.

[0038] The method for performing chromosome preparation of tetraploid petunia pistil stigma cells comprises the following steps:

...

Embodiment 3

[0044] Embodiment 3: basically the same as Embodiment 1, the difference is step (2), step (3) and step (4):

[0045] Step (2): Place diploid petunia young buds separately in 0.002mol.L -1 8-Hydroxyquinoline aqueous solution was pretreated at 6°C and protected from light for 3 hours, rinsed with distilled water 5 times with absorbent paper, and then transferred to Carnot’s fixative for 12 hours. Rinse and transfer to 70% ethanol for storage at 4°C for later use;

[0046] Step (3): Rinse the spare young buds in step (2) with distilled water for 5 times, then blot the water with absorbent paper, and then rinse with 0.075mol.L -1 Potassium chloride solution was hypotonic for 10 minutes at 1 mol.L -1 HCl solution was dissociated in a water bath at 60°C for 5 minutes, rinsed with distilled water for 5 times, then hypotonic with distilled water for 30 minutes, and blotted dry with absorbent paper.

[0047] (4) Transfer the flower buds after the low-osmosis treatment in step (3) i...

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Abstract

The invention discloses a flaking method of petunia chromosome. The method comprises the following steps: getting a petunia young bud at 8:00-10:00 in the morning of a clear day, placing in 0.002mol.L<-1> 8-hydroxyquinoline aqueous solution for pre-treating at low temperature, transferring into a Carnoy's fluid to be fixed, performing low-permeation through 0.075mol.L<-1> potassium chloride, and then water-bathing at 60 DEG.C in 1mol.L<-1> HCl solution for decomposing, washing and then low-permeating through the distilled water, and staining through aceto-carmine staining solution, transferring the young bud onto a glass slide, cutting to get a pistil stigma part, flaking and performing microscopy. The bud size, the pretreatment method, the decomposition and staining method and time and the flaking processes of petunias with different ploidy at optimal sampling time are determined, the obtained chromosome is uniform in dispersion, easy to count and easy for performing karyotype analysis, the operation is simple and convenient, the efficiency is high, and the method has a good application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for producing petunia chromosomes. Background technique [0002] Petunias ( Petunia hybrida Vilm.) is native to South America, also known as petunia, a plant of the genus Petunia in the family Solanaceae. It has huge flowers, rich colors, and a lot of flower patterns. It has long been the main potted flower and decorative plant in the world, and is widely used in The landscaping decoration and greening of courtyards, open-air flower beds and squares is known as "the king of flower beds in the world". Petunias are widely cultivated in the United States, and are often used in window sill beautification and urban landscape arrangement. Its production scale and quantity rank second among flower beds and garden plants in the United States; in European countries such as Italy, France, Spain, the Netherlands and Germany, Petunias are widely used in street beautificati...

Claims

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Application Information

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IPC IPC(8): G01N1/28
Inventor 魏跃嵇怡樊开青刘艳张莹史红林陈啸寅颜志明王全智
Owner JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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