Method of preparing haploid cells of Althaea rosea
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A haploid cell and hollyhock technology, applied in plant cells and other directions, can solve the problem of low haploid frequency, and achieve the effects of good repeatability, good growth and fast induction speed.
Inactive Publication Date: 2016-02-17
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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However, the frequency of naturally occurring haploids in nature is generally very low, only between 0.002% and 0.02%. According to the totipotency of plant cells, the use of anther culture to obtain haploid materials has successfully solved this problem. At present More than 200 plants with economic and ornamental value have obtained haploid through anther cultivation, most of which are Solanaceae and Poaceae, but there are few reports on anther cultivation of hollyhock and even Malvaceae plants
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Embodiment 1
[0017] Preparation of solid medium: Add 2,4-dichlorophenoxyacetic acid, naphthaleneacetic acid, 6-benzylaminopurine, sucrose and agar to MS medium so that the final concentration of 2,4-dichlorophenoxyacetic acid is 1mg / L , the final concentration of naphthaleneacetic acid is 0mg / L, the final concentration of 6-benzylaminopurine is 1.5mg / L, the final concentration of sucrose is 30g / L, the final concentration of agar is 6g / L, adjust pH=5.8, at 121°C, Sterilize under the condition of 0.1Mpa pressure for 25 minutes, cool to room temperature, and set aside.
Embodiment 2
[0019] Preparation of solid medium: Add 2,4-dichlorophenoxyacetic acid, naphthaleneacetic acid, 6-benzylaminopurine, sucrose and agar to MS medium so that the final concentration of 2,4-dichlorophenoxyacetic acid is 1mg / L , the final concentration of naphthaleneacetic acid is 1mg / L, the final concentration of 6-benzylaminopurine is 1.5mg / L, the final concentration of sucrose is 30g / L, the final concentration of agar is 6g / L, adjust the pH=5.8, at 121°C, Sterilize under the condition of 0.1Mpa pressure for 25 minutes, cool to room temperature, and set aside.
Embodiment 3
[0021] Preparation of solid medium: Add 2,4-dichlorophenoxyacetic acid, naphthaleneacetic acid, 6-benzylaminopurine, sucrose and agar to MS medium so that the final concentration of 2,4-dichlorophenoxyacetic acid is 1mg / L , the final concentration of naphthaleneacetic acid is 2mg / L, the final concentration of 6-benzylaminopurine is 1.5mg / L, the final concentration of sucrose is 30g / L, the final concentration of agar is 6g / L, adjust the pH=5.8, at 121°C, Sterilize under the condition of 0.1Mpa pressure for 25 minutes, cool to room temperature, and set aside.
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Abstract
The invention belongs to the field of biological engineering and relates to a method of preparing haploid cells of Althaea rosea. By using the totipotency principle of plant cells, the plant tissue culture technology and flow cytometry, the anthers of Althaea rosea that are sterilized are used as explants dispersely inoculated to a callus inducing medium, a proper hormone combination is obtained by screening, culturing is performed under proper conditions, calli are induced in about 20 d, ploidy of callusing cells is obtained by determination using a flow cytometer after 3-5 times of subculture, and the existence of callusing haploid cells is discovered. The invention makes first verification that it is probable to obtain haploid cells of Althaea rosea using anther culture and screens hormone combinations of high inducing rate. The inducing process is simple, the cost is low, ploidy identification by flow cytometer is fast and reliable, a new way is provided for haploid breeding of Althaea rosea and culturing of new species, and basis is laid for the kinetic study on the suspension culture of plant haploid cells.
Description
technical field [0001] The invention relates to a method for preparing plant haploid material, in particular to a method for preparing hollyhock haploid material. Background technique [0002] Hollyhock, also known as Yizhanghong, is a perennial herb of the genus Hollyhock in the Malvaceae family. The whole plant of hollyhock is used as medicine, which has great medicinal value, and has the effects of clearing away heat and detoxifying, antitussive and diuretic. Hollyhock contains a large amount of anthocyanins, which can be used as food coloring agents. The flowers of hollyhock are single or double-petal at the top of the raceme, with purple, pink, red, white and other colors. The color is bright and fresh, and has great ornamental value. It can be planted in courtyards and parks. It has strong resistance to hydrogen chloride, and is an excellent urban landscape plant. Famous varieties such as Chiba, Wuxin, Chongtai, Shearing, and Saw have been cultivated. Due to its exc...
Claims
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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04
CPCC12N5/04
Inventor 李兴林别振宇汪珊英
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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