A culture method for inducing asparagus microspores to obtain embryoid bodies

A culture method and microspore technology are applied in the field of culture of asparagus microspores to obtain embryoid bodies, which can solve the problems of high breeding difficulty, long breeding cycle, late start of asparagus breeding and the like, saving manpower and high seedling rate. , the effect of simple operation

Inactive Publication Date: 2014-10-15
JIANGXI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For a long time, because asparagus has the characteristics of dioecious plants, highly heterozygous genotypes, and perennials, its breeding is very difficult and the breeding cycle is very long.
In addition, asparagus is an imported product. my country's asparagus breeding started late, and foreign varieties have monopolized the Chinese market for a long time. The lack of new asparagus varieties with independent intellectual property rights has become one of the main bottlenecks restricting the development of my country's asparagus industry.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] (1) Preparation of culture medium and solid medium: the basic medium used in the two mediums is MS, and the carbon source used is sucrose; the coagulant added to the solid medium is agar, and its mass concentration is 0.7%; Liquid is in MS, add 60g. L -1 sucrose, 0.4 mg. L -1 6-benzylaminoadenine 6-BA, 0.5 mg. L -1 2,4-dichlorophenoxyacetic acid 2,4-D, 0.1mg﹒ L -1 Naphthaleneacetic acid NAA, 0.03mg. L -1 Glutathione L-GLU, pH5.8; the composition of the solid medium is: in MS medium, add 7g. L -1 of agar and 30 g. L -1 Sucrose, pH5.8;

[0010] (2) Sterilization and subpackage of the medium: solid culture is based on sterilization at 121 °C and 1.1 MPa for 20 minutes, and the culture solution is sterilized by filtration with a plastic filter, which is equipped with a 0.22 micron microporous membrane to ensure the culture The aseptic state of the solution; after that, divide the packaging, inject 10 mL of solid medium into each Petri dish before solidification...

Embodiment 2

[0020] (1) Preparation of culture medium and solid medium: the basic medium used in the two mediums is MS, and the carbon source used is sucrose; the coagulant added to the solid medium is agar, and its mass concentration is 0.75%; Liquid is in MS, add 70g. L -1 sucrose, 0.5 mg. L -1 6-benzylaminoadenine 6-BA, 1.0 mg. L -1 2,4-dichlorophenoxyacetic acid 2,4-D, 0.5mg. L -1 Naphthaleneacetic acid NAA, 0.03mg. L -1 Glutathione L-GLU, pH5.8; the composition of the solid medium is: in MS medium, add 7.5 g. L -1 of agar and 30 g. L -1 sucrose, pH5.8;

[0021] (2) Sterilization and subpackage of the medium: solid culture is based on sterilization at 121 °C and 1.1 MPa for 20 minutes, and the culture solution is sterilized by filtration with a plastic filter, which is equipped with a 0.22 micron microporous membrane to ensure the culture The aseptic state of the solution; after that, divide the packaging, inject 10 mL of solid medium into each Petri dish before solidifica...

Embodiment 3

[0031] (1) Preparation of culture medium and solid medium: the basic medium used in the two mediums is MS, and the carbon source used is sucrose; the coagulant added to the solid medium is agar, and its concentration is 0.8%; the culture medium is prepared That is, in MS, add 70g. L -1 sucrose, 0.8 mg. L -1 6-benzylaminoadenine 6-BA, 1.0 mg. L -1 2,4-dichlorophenoxyacetic acid 2,4-D, 1.0 mg. L -1 Naphthaleneacetic acid NAA, 0.03 mg. L -1 Glutathione L-GLU, pH5.8; the composition of the solid medium is: in MS medium, add 8.0 g. L -1 of agar and 30 g. L -1 sucrose, pH5.8;

[0032] (2) Sterilization and subpackage of the medium: solid culture is based on sterilization at 121 °C and 1.1 MPa for 20 minutes, and the culture solution is sterilized by filtration with a plastic filter, which is equipped with a 0.22 micron microporous membrane to ensure the culture The aseptic state of the solution; after that, divide the packaging, inject 10 mL of solid medium into each Pe...

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Abstract

The invention discloses a culture method of an embryoid by inducing an asparagus microspore, which belongs to the field of plant biological engineering. The method comprises the following steps of: preparing a culture medium, sterilizing the culture medium, selecting a plant and a bud, determining the development period of the microspore, pretreating and disinfecting the bud, stripping, inoculating and culturing anther, and automatically dissociating and developing the microspore to form the embryoid, wherein the obtained embryoid of the microspore can normally grow, and seedling number is large. By adopting the method disclosed by the invention, 7-8 weeks are generally needed from the beginning of the culture to the formation of the embryoid; haploid or double haploid can be obtained; the double haploid is widely known as a good breeding material and can be applied to the seed selection of a new asparagus type and the modification of the asparagus type; if the culture method is applied to a breeding practice, the breeding period can be greatly reduced, selection efficiency is greatly improved, and a lot of manpower and materials are saved. and therefore the culture method is a quite effective method for accelerating the pure line seed selection of asparagus breeding.

Description

technical field [0001] The invention relates to a culture method for inducing asparagus microspores to obtain embryoid bodies. Background technique [0002] Asparagus (Asparagus officinalis L.), also known as Asparagus officinalis L., is a perennial herbaceous plant of Asparagus asparagus, diploid (2X=20), and is an important functional vegetable. Tumor, lowering blood pressure, lowering blood fat, lowering blood sugar and other effects, enjoy the reputation of "king of vegetables". At present, my country's asparagus industry is developing rapidly, and its planting and sales volume ranks first in the world. For a long time, because asparagus has the characteristics of dioecious plants, highly heterozygous genotypes, and perennials, its breeding is very difficult and the breeding cycle is very long. In addition, asparagus is an imported product. my country's asparagus breeding started late, and foreign varieties have monopolized the Chinese market for a long time. The lack ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 陈光宇汤泳萍罗绍春周劲松张岳平谢启鑫管珊红
Owner JIANGXI ACAD OF AGRI SCI
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