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Analysis method of sharp leaf CinnamomumKanehirae Hay karyotype based on root tips

A technology of chromosome karyotype and analysis method, which is applied in the field of root tip-based analysis of the chromosome karyotype of Cinnamomum chinensis to achieve the effect of easy preservation

Inactive Publication Date: 2019-09-10
广州甘蔗糖业研究所湛江甘蔗研究中心
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, there is no report on the karyotype analysis of Cinnamomum camphora

Method used

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  • Analysis method of sharp leaf CinnamomumKanehirae Hay karyotype based on root tips
  • Analysis method of sharp leaf CinnamomumKanehirae Hay karyotype based on root tips
  • Analysis method of sharp leaf CinnamomumKanehirae Hay karyotype based on root tips

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Effects of different pretreatment methods on the effect of chromosome preparation

[0039] The root tips of Cinnamomum camphora tissue culture seedlings were collected from 8:00 to 10:00 in the morning on a sunny day, and three pretreatment methods were used for pretreatment experiments. After rinsing with water, transfer to Carnot's fixative solution (absolute ethanol: glacial acetic acid = 3:1), fix for 24 h, and store in 70% ethanol at 4°C for later use. According to the method of enzymatic hydrolysis to remove the wall and hypotonicity, the chromosome specimens were prepared by fixation, pre-hypotonicity, enzymatic hydrolysis, post-hypotonicity, post-fixation and smear, then stained with Giemsa, observed with a POTIKA upright microscope, and photographed by the Optika Vision pro camera system .

[0040] The results of 3 different pretreatment methods on the root tips of Cinnamomum camphora showed that (Table 1): Different pretreatment methods had certain effects on...

Embodiment 2

[0044] Effect of different enzymatic hydrolysis time on the effect of chromosome production

[0045] Collect the root tips of Cinnamomum camphora tissue culture seedlings from 9:00 to 11:00 in the morning on a sunny day, place them in 0.002mol / L 8-hydroxyquinoline solution for pretreatment for 2 hours, rinse with water and transfer to Carnot’s fixative solution (absolute ethanol: Glacial acetic acid = 3:1), fixed for 24 hours, and stored in 70% ethanol at 4°C in a refrigerator for later use. Chromosome specimens were prepared by pre-hypotonicity, enzymatic hydrolysis (3 time gradients), post-hypotonicity, post-fixation and smear according to the method of enzymatic hydrolysis to wall hypotonicity, then stained with Giemsa, and observed with POTIKA upright microscope, Optika Vision The pro camera system takes pictures. The results of the effect of different enzymatic hydrolysis time on the effect of Cinnamomum camphora chromosome production showed that ( figure 1): The effect...

Embodiment 3

[0047] Chromosome Karyotype Analysis of Cinnamomum camphora

[0048] Collect the root tips of Cinnamomum camphora tissue culture seedlings from 9:00 to 11:00 in the morning on a sunny day, place them in 0.002mol / L 8-hydroxyquinoline solution for pretreatment for 2 hours, rinse with water and transfer to Carnot’s fixative solution (absolute ethanol: Glacial acetic acid = 3:1), fixed for 24 hours, and stored in 70% ethanol at 4°C in a refrigerator for later use. Chromosome specimens were prepared by pre-hypotonicity, enzymatic hydrolysis, post-hypotonicity, post-fixation and smear according to the method of enzymatic hydrolysis to remove the wall and hypotonicity, and then stained with Giemsa, observed with a POTIKA upright microscope, and photographed by the Optika Vision pro camera system.

[0049] Arm ratio (r) = long arm (S) / short arm (L)

[0050] Chromosome relative length (%) = chromosome length / total length of chromosome group × 100%

[0051] Average length karyotype ...

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Abstract

The invention discloses an analysis method of sharp leaf CinnamomumKanehirae Hay karyotype based on root tips. The method comprises the following steps: collecting a robust, white and tender root tipof a tissue culture seedling of CinnamomumKanehirae Hay in a sunny morning to serve as a test material, pretreating, fixing and storing the test material, performing fixation, anterior hyposmosis, enzymolysis, post hyposmosis and smearing by using an enzymolysis wall removal hyposmosis method to obtain a better chromosome specimen, wherein the enzymolysis is performed by using a mixture of 4% cellulase and 5% pectinase, and performing Giemsa staining and microscopic examination. The method is simple and easy to operate, is suitable for the analysis method of sharp leaf CinnamomumKanehirae Haykaryotype with in-situ PCR, FISH operations, and provides a technical basis for the cytological study of the CinnamomumKanehirae Hay. The karyotype analysis method and the karyotype of the sharp leafCinnamomumKanehirae Hay are reported in the invention for the first time, which indicates that the CinnamomumKanehirae Hay has 24 chromosomes, the karyotype formula is 2n=2x=24=22m+2sm, the asymmetrycoefficient of the karyotype is 55.81%, and the karyotype belongs to a type 2B and belongs to relatively primitive species. A certain theoretical basis is provided for the origin, evolution and genetic breeding of the sharp leaf CinnamomumKanehirae Hay.

Description

technical field [0001] The invention relates to a chromosome karyotype analysis method, in particular to a root tip-based chromosome karyotype analysis method of Cinnamomum camphorii, belonging to the field of tree cytology research technology. Background technique [0002] Cinnamomum Kanehirae Hay belongs to the genus Lauraceae, also known as black camphor, which is a native evergreen broad-leaved tree species in Taiwan, China. Cinnamomum camphora has high economic value. The whole plant of roots, stems and leaves can be used to extract the essential oil of Cinnamomum camphora. Wood is the best material for carving furniture. In addition, Antrodia camphorata is the only host of Antrodia camphorata. Antrodia camphorata is known as the "king of medicinal mushrooms" in medicine. It is often used to sober up and relieve fatigue, and has immune regulation and liver protection effects. [0003] The research on Cinnamomum camphora started late, and the research direction was sing...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/84G01N15/14G01N1/28G01N1/30
CPCG01N1/2813G01N1/30G01N15/14G01N21/84G01N2015/1006G01N2015/1486
Inventor 官锦燕罗青文谭嘉娜罗剑飘黄海英文明富
Owner 广州甘蔗糖业研究所湛江甘蔗研究中心
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