Method for identifying carnation chromosome number by bud

Abstract

The invention provides a method for identifying carnation chromosome number by bud, comprising: obtaining bud anther, ovary wall and corolla tissue, pre-treating ice water mixture, fixing by stationary liquid, dyeing, dissociating with hydrochloric acid, tabletting and microscopic examining. Compared with the traditional method that a root tip tissue is used for chromosome tabletting, the invention solves the disadvantages of few visual filed cells, difficult microscopic examination and poor chromosome dispersion during root tip chromosome tabletting, and overcomes the problems of long rootage period for carnation cutting seedling and time and labor wasting for cutting; compared with stem tip chromosome tabletting, the invention overcomes the interference problem of non-meristematic cell or reserve substance or secretion, chromosome tabletting can be realized only by using buds, which does not damage plant growing state and has convenient material resource; in addition, a plurality ofmetaphase cells exist, so that the difficulties of small carnation chromosome, huge quantity and hard counting of the chromosome can be solved. Thus, when plants bloom, the effective method is that the bud is used for chromosome tabletting.

Description

technical field

[0001] The invention relates to a method for identifying the number of carnation chromosomes, in particular to a method for identifying the number of carnation chromosomes by flower buds, and belongs to the technical fields of cell biology and cytogenetics. Background technique

[0002] Carnation (Dianthus caryophyllus Linn), also known as carnation, is a perennial herbaceous plant of the genus Caryophyllus in the Caryophyllaceae family. It is one of the world-renowned fresh cut flowers and one of the most important bulk export flowers in Yunnan Province. In the hybrid breeding of carnation, there are hybrid incompatibility and low seed setting rate. One reason for this is that the cytological background of many accessions is unclear. Therefore, in order to increase the success rate of hybridization and improve the efficiency of hybrid breeding, the problem of chromosome ploidy analysis of Carnation should be solved first.

[0003] Chromosomes are the carri...

Examples

Embodiment 1

[0019] Using carnation variety "Autumn Harvest" as material, the 1.9cm-long flower buds were picked, the ovary was stripped, the ovary was cut longitudinally, and the ovary wall was pretreated in ice-water mixture for 24 hours; the waste was sucked with a 5ml syringe After liquid solution, the pretreated ovary wall was washed 3 times with distilled water, immersed in Carnot's I fixative solution with a volume ratio of absolute ethanol: glacial acetic acid = 3:1, and fixed for 2 hours; after absorbing the waste liquid with the same syringe, , wash the fixed ovary wall three times with distilled water, immerse it in the HCL solution with a concentration of 1mol / L for 20min at room temperature, and dissociate it with hydrochloric acid; use the same syringe to absorb the waste liquid, and then wash the ovary wall with distilled water 3 times, put it on the glass slide, tear up the ovary wall tissue with a scalpel, drop 2 drops of carbo fuchsin staining solution, stain for 10 minute...

Embodiment 2

[0021] Using carnation variety "Huang Feihong" as material, pick 1.3cm-long flower buds, peel off the ovary, cut the ovary longitudinally, take the ovary wall and put it in ice-water mixture for 22 hours; use a 5ml syringe to absorb After treating the waste liquid, wash the ovary wall twice with distilled water, and then immerse it in Carnot's I fixative with a volume ratio of absolute ethanol:glacial acetic acid=3:1 and fix it for 4h; After the waste liquid was collected, the ovary wall was washed 3 times with distilled water, immersed in the HCL solution with a concentration of 1mol / L at room temperature for 15 minutes, and dissociated with hydrochloric acid; after the treated waste liquid was absorbed with the same syringe, the ovary wall Then wash with distilled water 3 times, put it on the glass slide, tear up the ovary wall tissue with a scalpel, drop 3 drops of carbo fuchsin staining solution, stain for 5 minutes, cover with a cover glass, cover with absorbent paper, and...

Embodiment 3

[0023] Using the carnation variety "Obsession" as the material, take 1.5cm-long flower buds. At this time, the pollen is in the single-nucleus stage, take out the anthers, and put them in the ice-water mixture for pretreatment for 24 hours; use a 5ml syringe to absorb the treated waste liquid. , the ovary wall was cleaned 3 times with distilled water, immersed in Carnot’s I fixative with a volume ratio of absolute ethanol:glacial acetic acid=3:1 and fixed for 2h; after absorbing the treated waste liquid with the same syringe, the ovary was The wall of the ovary was washed 3 times with distilled water, immersed in HCL solution with a concentration of 1mol / L for 20 min at room temperature, and dissociated with hydrochloric acid; after absorbing the treated waste liquid with the same syringe, the wall of the ovary was washed 3 times with distilled water, and placed On the glass slide, tear up the anther tissue with a scalpel, drop 2 drops of carbo fuchsin staining solution, stain ...

technical field

[0001] The invention relates to a method for identifying the number of carnation chromosomes, in particular to a method for identifying the number of carnation chromosomes by flower buds, and belongs to the technical fields of cell biology and cytogenetics. Background technique

[0002] Carnation (Dianthus caryophyllus Linn), also known as carnation, is a perennial herbaceous plant of the genus Caryophyllus in the Caryophyllaceae family. It is one of the world-renowned fresh cut flowers and one of the most important bulk export flowers in Yunnan Province. In the hybrid breeding of carnation, there are hybrid incompatibility and low seed setting rate. One reason for this is that the cytological background of many accessions is unclear. Therefore, in order to increase the success rate of hybridization and improve the efficiency of hybrid breeding, the problem of chromosome ploidy analysis of Carnation should be solved first.

[0003] Chromosomes are the carri...

Examples

Embodiment 1

[0019] Using carnation variety "Autumn Harvest" as material, the 1.9cm-long flower buds were picked, the ovary was stripped, the ovary was cut longitudinally, and the ovary wall was pretreated in ice-water mixture for 24 hours; the waste was sucked with a 5ml syringe After liquid solution, the pretreated ovary wall was washed 3 times with distilled water, immersed in Carnot's I fixative solution with a volume ratio of absolute ethanol: glacial acetic acid = 3:1, and fixed for 2 hours; after absorbing the waste liquid with the same syringe, , wash the fixed ovary wall three times with distilled water, immerse it in the HCL solution with a concentration of 1mol / L for 20min at room temperature, and dissociate it with hydrochloric acid; use the same syringe to absorb the waste liquid, and then wash the ovary wall with distilled water 3 times, put it on the glass slide, tear up the ovary wall tissue with a scalpel, drop 2 drops of carbo fuchsin staining solution, stain for 10 minute...

Embodiment 2

[0021] Using carnation variety "Huang Feihong" as material, pick 1.3cm-long flower buds, peel off the ovary, cut the ovary longitudinally, take the ovary wall and put it in ice-water mixture for 22 hours; use a 5ml syringe to absorb After treating the waste liquid, wash the ovary wall twice with distilled water, and then immerse it in Carnot's I fixative with a volume ratio of absolute ethanol:glacial acetic acid=3:1 and fix it for 4h; After the waste liquid was collected, the ovary wall was washed 3 times with distilled water, immersed in the HCL solution with a concentration of 1mol / L at room temperature for 15 minutes, and dissociated with hydrochloric acid; after the treated waste liquid was absorbed with the same syringe, the ovary wall Then wash with distilled water 3 times, put it on the glass slide, tear up the ovary wall tissue with a scalpel, drop 3 drops of carbo fuchsin staining solution, stain for 5 minutes, cover with a cover glass, cover with absorbent paper, and...

Embodiment 3

[0023] Using the carnation variety "Obsession" as the material, take 1.5cm-long flower buds. At this time, the pollen is in the single-nucleus stage, take out the anthers, and put them in the ice-water mixture for pretreatment for 24 hours; use a 5ml syringe to absorb the treated waste liquid. , the ovary wall was cleaned 3 times with distilled water, immersed in Carnot’s I fixative with a volume ratio of absolute ethanol:glacial acetic acid=3:1 and fixed for 2h; after absorbing the treated waste liquid with the same syringe, the ovary was The wall of the ovary was washed 3 times with distilled water, immersed in HCL solution with a concentration of 1mol / L for 20 min at room temperature, and dissociated with hydrochloric acid; after absorbing the treated waste liquid with the same syringe, the wall of the ovary was washed 3 times with distilled water, and placed On the glass slide, tear up the anther tissue with a scalpel, drop 2 drops of carbo fuchsin staining solution, stain ...