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Method for quickly identifying chromosome number of rosa plant

A technology of chromosome number and rose genus, which is applied in the field of plant cell biology and cytogenetics, can solve the problems of limited number of mitotic cells, difficulty in seed germination, inability to count, etc., to achieve accurate size and position of the sample, and shorten the pretreatment time , Time is easy to control the effect

Inactive Publication Date: 2014-04-09
CHINA AGRI UNIV
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AI Technical Summary

Problems solved by technology

However, because Rosa plants are woody, and the genetic background is complex, the chromosomes are small and medium-sized, which makes chromosome counting and analysis difficult, and the method for identifying the number of chromosomes of herbaceous plants such as Carnation cannot be effectively applied to Rosa plants
In addition, Rosa plants are not easy to obtain suitable materials, and the tissue cells of the materials are difficult to dissociate, resulting in inaccurate, unstable or even impossible counting of chromosomes, and the counting time is long, which is not suitable for fast and accurate chromosome numbering of large-scale varieties. identification
[0005] The material commonly used to identify plant chromosomes is the root tip, but it is difficult for the seeds of Rosa plants to germinate, and there are variations in the seeded offspring, especially the rose (Rosa Hybrida)
In addition, it is difficult for many varieties or varieties of Rosa to take root from cuttings, the cutting cycle is long, time-consuming and laborious, and the adventitious roots produced are easy to lignify, and it is difficult to grasp the time of material collection. In addition, when doing microscopic examination, there are few mitotic cells in the field of view of the microscope, and the dispersion of chromosomes is poor. Chromosome count
Although it has been reported that the young stem tips of Rosa plants were used as materials, it is difficult to obtain accurate materials due to the limited number of mitotic cells at the growth point of the shoot tips, and there are pressures from the difficult dissociation of tissue cells and the interference of cell inclusions or secretions. Film effects and other issues
The number of chromosomes can be identified by flower buds on Carnation, but there is no report on the use of flower buds of Rosa plants as materials for chromosome count analysis

Method used

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  • Method for quickly identifying chromosome number of rosa plant
  • Method for quickly identifying chromosome number of rosa plant
  • Method for quickly identifying chromosome number of rosa plant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] The diploid young flower buds of R. chinensis 'Old Pink Daily'. are used as materials.

[0076] A. At 9:00 in the morning, take the tops of the branches in the flowering stage cultivated in the open field, and the young flower buds with a round and compact external shape, and quickly strip the materials with a height of about 2.0mm under the stereoscope.

[0077] B. Use p-dichlorobenzene at 25°C for 3 hours. The top of the young flower buds is the unexpanded sepal, which is densely covered with fluff. Before processing, use a scalpel to cut off the top 0.5mm under the stereoscope, put it into 20ml of saturated p-dichlorobenzene solution, and use a needle The syringe was vacuumed until the material sank to the bottom of the pretreatment solution, and then washed 5 times with deionized water.

[0078] C. Use Carnot's fixative solution to fix in an ice box for 5 hours, and then wash with deionized water for 5 times.

[0079] D. Dissociate with 1M hydrochloric acid at 60°...

Embodiment 2

[0084] The stem tip of diploid R. chinensis 'Old Pink Daily'. is used as the material.

[0085] At 9:00 in the morning, take the tops of vigorously growing annual branches cultivated in the open field, and quickly strip off stems with a length of about 1.5 mm under a stereoscope. Treat in a saturated solution of p-dichlorobenzene at 25°C for 2.5 hours, wash with deionized water 5 times; dissociate with 1M hydrochloric acid for 8 minutes. Both the first and second strokes are carried out with a force of 28N and a frequency of 80; the rest of the steps are the same as those described in case (1). For the results, please refer to image 3 b.

Embodiment 3

[0087] The shoot tip of diploid Baiyutang (R.multiflora var.albo-plena) was used as material.

[0088]At 9:00 in the morning, take the top 2cm of the annual young shoots cultivated in the open field, peel off the surrounding leaves layer by layer under the stereoscope to expose the top of the stem, and use a scalpel to cut off the top growth point with a length of about 2.5mm at the front . The saturated solution of p-dichlorobenzene was treated at 25°C for 3.0h. The leaves of the newly grown young terminal buds of this material are densely covered with white hairs. Carnot's fixative was fixed for 4 hours, and 1M hydrochloric acid aqueous solution was dissociated for 9 minutes. Both the first and second percussions were performed with a force of 34N and a number of times of 100. The remaining steps are the same as those described in case (1), and the results can be found in image 3 E and 3H. It can be seen from the results that there is a deletion problem in the diploid ...

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Abstract

The invention provides a method for quickly identifying chromosome number of a rosa plant. The method comprises the steps of drawing materials, pretreatment, fixing, disaggregating, dyeing, tabletting, microscopic examination and the like. The tabletting process is digitalized in the tabletting step, and cover glass on the material is vertically knocked on by 28-34N of force for 160-200 times for the best. In preferable embodiments, the stem tip and / or blossom bud are adopted as materials for counting and analyzing the chromosomes. In preferable embodiments, the materials are obtained between nine and eleven in the morning or between three and four in the afternoon. In embodiments, growth points and positions of the materials are confirmed by an electron microscope in a scanning manner. The method disclosed by the invention has the advantages of diversified materials, accurate tabletting and the like, and is quick, accurate and efficient, and is especially applicable to quick and accurate identification of the chromosome number of the rosa plants on a large scale. Thus, the method is applied to quick and accurate identification of the chromosome number of the rosa plants on a large scale.

Description

technical field [0001] The invention relates to a method for identifying the number of chromosomes of plants, especially plants of the genus Rosacea, and belongs to the fields of plant cell biology and cytogenetics. technical background [0002] Chromosomes are the carriers of genes, and the preparation of chromosome specimens is the basis of cytogenetics. Excellent chromosome preparation technology is also a prerequisite for chromosome banding, karyotype analysis, and in situ hybridization. The observation and analysis of plant cell chromosomes is of great significance for the analysis of karyotype analysis, the analysis of chromosome behavior during meiosis, the analysis of chromosome behavior and cell variation during cell fusion culture, and other genetic phenomena. In addition, the variation produced by genetic engineering breeding at the gene level can also be located by chromosome banding, in situ hybridization, and fluorescence in situ hybridization based on staining...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCG01N1/30
Inventor 赵梁军寇亚平赵宏伟高彬
Owner CHINA AGRI UNIV
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