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Tabletting method of eucalyptus chromosome

A chromosome and tablet technology, which is applied in the field of plant identification, can solve the problems of lack of mature, stable and high-quality chromosome tablet technology, little research at the cell level, and high price of colchicine, so as to reduce the degree of toxicity and risk, reduce experiment cost and improve stability

Inactive Publication Date: 2016-06-22
RES INST OF TROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Eucalyptus is an exotic tree species and an important plantation tree species in southern my country. The current research on it mainly focuses on introduction and cultivation, wood processing, genetic improvement, genetic map construction and molecular marker-assisted breeding, high-efficiency cultivation and ecological effects. Little research has been done on the cellular level
Some people abroad have done observation and research on the chromosomes of different species of eucalyptus, but they have the disadvantages of poor stability and low success rate; therefore, sometimes it is necessary to prepare a large number of samples to obtain clearly visible karyotype analysis images; because colchicine is expensive, Lead to higher cost of tableting
There are only sporadic studies on eucalyptus chromosome compression technology in China. From the research reports, there is still no mature, stable and high-quality chromosome compression technology

Method used

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  • Tabletting method of eucalyptus chromosome
  • Tabletting method of eucalyptus chromosome
  • Tabletting method of eucalyptus chromosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Get the root tip of the diploid tissue-cultured rooted seedling of Eucalyptus macrophylla clone DH32-29, put it into 0.01g / L amisulfurin and soak it for 4h, then wash the root tip after soaking with distilled water, put it in a card Fix in Knott's fixative solution at 4°C for 24h, take out the root tip from the fixative solution, rinse with distilled water, put 10g / L cellulase and 10g / L pectinase at a volume ratio of 1:1 at 37°C Dissociate in the mixed solution for 30 minutes, take out the root tip, wash it with distilled water, put it on a clean glass slide, cut off the root cap and extension part with a razor blade, drop 1 drop of carbo fuchsin staining solution, and stain for 10 minutes. Cover with a cover slip, gently tap the cover slip with an eraser, blot the excess staining solution with absorbent paper, and then place the pressed slide under a microscope for observation. Chromosome observation map as figure 1 shown.

Embodiment 2

[0035] The enzymolysis time in embodiment 1 is increased to 40min by 30min, all the other operations are all the same as embodiment 1. Chromosome observation map as figure 2 shown.

Embodiment 3

[0037] The enzymolysis time in Example 1 was increased from 30min to 35min, and all the other operations were the same as in Example 1. The obtained chromosome observation map is as follows image 3 shown.

[0038] From Figure 1~3 It can be seen that when the enzymatic hydrolysis is 30min (such as figure 1 ), the enzymatic hydrolysis was insufficient, and the chromosomes were still bound in the nucleus without being dispersed; when the enzymatic hydrolysis was 40min (such as figure 2 ), excessive dissociation, not only the cell membrane is decomposed, but also the chromosomes are decomposed, and it is difficult to observe the formed chromosomes. Only when the dissociation time is appropriate can the ideal dissociation effect be obtained. Therefore, considering the convenience and cost of operation, it is better to dissociate the mixed solution of cellulase and pectinase at a constant temperature of 37°C for 35min (such as image 3 ).

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Abstract

The invention belongs to the technical field of plant identification and discloses a tabletting method of eucalyptus chromosome.The tabletting method includes following steps: taking materials, pretreating, fixing, dissociating, dyeing, tabletting and performing microscopic examination.In the tabletting method, oryzalin is adopted as a pretreatment agent, so that cytotoxic effect is reduced obviously when cell mitosis is blocked; cells can be in normal mitosis, so that during fixing, number of accumulated metaphase cells is large, chromosome tabletting success probability is increased, and stability is improved.Oryzalin is low in cost and far lower than colchicine in toxicity; in the tabletting process, environment pollution can be reduced and degree and risk of causing damage to people are lowered.

Description

technical field [0001] The invention belongs to the technical field of plant identification, in particular to a method for pressing eucalyptus chromosomes. Background technique [0002] Chromosomes are the carrier and material basis of genetic material at the level visible to the naked eye. The number and shape of chromosomes are significant hereditary characteristics of a species, which can be used to identify or assist in the identification of species, the relationship between species and the degree of variation of species. It is a basic and basic cell biology research technology and means to master the technology of chromosome compression, and to observe the number and shape of chromosomes. It is widely used in cross breeding and other research. [0003] Eucalyptus is a general term for the tree species of the genus Angophora, Corymbia and Eucalyptus in the family Mytraceae. Eucalyptus is an exotic tree species and an important plantation tree species in southern my cou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/30
CPCG01N1/2813G01N1/30
Inventor 韩超徐建民
Owner RES INST OF TROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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