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Method for identifying single chromosome of winged tobaccos

A single-chromosome, identification method technology, applied in material excitation analysis, fluorescence/phosphorescence, instruments, etc., can solve the problems of difficult cell wall, difficult to accurately identify single chromosome, and high dispersion.

Pending Publication Date: 2020-06-23
GUIZHOU TOBACCO SCI RES INST +1
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Problems solved by technology

[0007] Due to the small size of tobacco chromosomes, it is difficult to fully dissociate the cell wall, and the above-mentioned techniques are difficult to obtain slices with many cleavage phases and high dispersion; the chromosomes are small and the morphological differences between homologous chromosomes are not obvious, and it is difficult to accurately identify single chromosomes only by morphological characteristics.

Method used

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  • Method for identifying single chromosome of winged tobaccos
  • Method for identifying single chromosome of winged tobaccos
  • Method for identifying single chromosome of winged tobaccos

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Embodiment Construction

[0023] The method for segregating and identifying single chromosomes of Nicotiana winga, the specific method is as follows:

[0024] (1) Chromosomal specimen preparation: soak the mature and plump seeds of Nicotiana winga in distilled water at 40°C for 50 minutes, sow them on damp filter paper, place them in a petri dish (put a piece of filter paper on the bottom of the petri dish to keep it moist), and incubate at 26°C germination. When the root grows to 0.5-1 cm, cut the fresh root tip between 9:00 and 11:00 in the morning, and put it into a saturated α-bromide solution for pretreatment at room temperature for 5 hours. Wash with distilled water three times, and fix overnight with Carnot's fixative solution (ethanol: glacial acetic acid = 3:1). After washing with distilled water for 3 times, hypotonic for 1 h before placing in distilled water, treat with 3% cellulose-pectinase mixed enzyme solution at 37°C for 2.5 h, wash 3 times, and hypotonic for 60 min after placing in di...

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Abstract

The invention discloses a method for identifying a single chromosome of winged tobaccos. The method comprises the following steps of (1) preparing a chromosome specimen: germinating mature and full seeds of the winged tobaccos, when roots grow to 0.5-1cm, clipping fresh root tips at 9: 00-11: 00 in the morning, putting the fresh root tips into a saturated alpha-bromo-naphthalene solution for room-temperature pretreatment, cleaning, carrying out low permeability and then putting into distilled water, treating with a cellulose-pectinase mixed enzyme solution, manufacturing chromosome preparations, and dyeing with a saturated acetic acid lichen red dye solution; (2) carrying out karyotype analysis, heterochromatin band spectrum analysis and 45s and 5s rDNA chromosome spectrum analysis on thechromosome preparations; and (3) identifying the single chromosome: comparing and analyzing a karyotype analysis result, a heterochromatin band spectrum and 45S and 5SrDNA chromosome site characteristics of each homologous chromosome of the winged tobaccos to finish accurate identification of each single chromosome of a chromosome group of the winged tobaccos.

Description

technical field [0001] The invention relates to a method for identifying a winged tobacco single chromosome, belonging to the field of biotechnology. Background technique [0002] Plant root tip cell mitosis and chromosome production technology: The meristematic tissues of vigorous growth of plants, such as root tip, shoot tip, and young leaves, are undergoing mitosis. Take these tissues as materials, kill the cells quickly after fixing, and keep the substances and living conditions in the cells, and then go through dissociation, staining, pressing and other processes to quickly disperse and attach the cells to the glass slide and Between the coverslips, a large number of cells in various stages of mitosis can be seen under the microscope, and the chromosomes can be observed. In metaphase, the centromeres of each chromosome are arranged on the equatorial plane, and the two arms extend to both sides of the equatorial plane. The spindle is fully formed, with one end attached...

Claims

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Application Information

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IPC IPC(8): G01N1/31G01N21/64
CPCG01N1/31G01N21/6428G01N21/6458G01N21/6486G01N2021/6439
Inventor 盛茂银王霖娇任学良
Owner GUIZHOU TOBACCO SCI RES INST
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