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Method for establishing prostrate spurge regeneration system through somatic embryogenesis way

A system, the technology of Dijin, which is applied in the field of plant cell engineering, can solve the problems of seasonal restrictions, limitations, and low regeneration rate of explants, and achieve the effects of uniform traits, normal plant development, and easy genetic engineering operations.

Inactive Publication Date: 2007-06-06
INST OF FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Although the brocade regeneration system has been established through the above methods, the regeneration rate is low, and the selection of explants is greatly restricted by the season, so it is greatly limited in practical application.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] In this experiment, the brocade seeds imported from Canada were used. Soak the brocade seeds in tap water at room temperature for 10 to 15 hours, then sterilize the surface with 70% alcohol for 2 minutes, rinse them with sterile water twice, and then rinse them with 0.1% chlorinated water. Mercury HgCl 2 Sterilize the surface for 10 minutes, rinse 5 times with sterile water, cut out the embryos with tweezers and a scalpel and inoculate them in the modified milk protein containing 6-BA 2.3mg / l, 2,4-D 3.8mg / l, 600mg / l hydrolyzed milk protein. B 5 culture medium (Table 1). The medium contains 30g / L sucrose and 6.5g / L agar. Use 1mol / L NaOH or HCl to adjust the pH to 5.8-6.2, sterilize with high pressure steam for 20 minutes, cultivate at 22-28°C, light-dark cycle 16h / 8h, light intensity 2000-2500lx, relative humidity 60%-70%, 30 days to replace the fresh medium.

[0023] The callus began to appear in 7 to 10 days, and the callus grew rapidly after 15 days. The callus i...

Embodiment 2

[0030] The culture procedures were all carried out according to Example 1, the difference was that in the first stage, 6-BA1.7mg / l, 2,4-D 4.3mg / l and hydrolyzed milk protein 700mg / l were added, and in the second stage, the first Subculture 2,4-D1.8mg / l, 6-BA 0.9mg / l, second subculture 2,4-D0.9mg / l, 6-BA 0.45mg / l, third subculture 2, 4-D 0.9mg / l, 6-BA 0.45mg / l, in the third stage, transfer the embryogenic callus obtained by subculture to B without adding growth regulators and adding 600mg / l hydrolyzed milk protein 5 performed on culture medium.

Embodiment 3

[0032] The culture procedures were all carried out as in Example 1, except that in the first stage, 6-BA 2.5 mg / l, 2,4-D 3.5 mg / l and hydrolyzed milk protein 800 mg / l were added, and in the second stage, the first Passage 2,4-D 2.5mg / l, 6-BA 1.25mg / l, second passage 2,4-D 1.25mg / l, 6-BA 0.65mg / l, third passage 2, 4-D1.25mg / l, 6-BA 0.65mg / l, in the third stage, the embryogenic callus obtained by subculture was transferred to B without growth regulators and added 460mg / l hydrolyzed milk protein 5 performed on culture medium.

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Abstract

The invention relates a method of establishing regenerative system with using mature stock as material, comprising the three steps: (1) culturing the mature stock to evoke callus; (2) subculturing the callus; (3) the somatic embryo produced in the callus and developing to plant. The generation rate of somatic embryo is rised to 53% due to adopt the method of fractional reducing the concentration of 2, 4-dichlorophenoxyacetic acid in successive transfer cultureto,

Description

technical field [0001] The invention relates to a method for establishing a regeneration system for in vitro culture of Radix chinensis. Specifically, it relates to a method for establishing a regeneration system through the somatic embryogenesis pathway by taking the mature embryo of the brocade seed as an explant. It belongs to the field of plant cell engineering. Background technique [0002] Brocade belongs to the genus Parthenocissus of the family Vitaceae, also known as creeper. It is a deciduous vine with strong adsorption and climbing capabilities. Brocade is a garden, urban vertical greening and landscaping plant with good ornamental effect. The United Nations Biosphere and Environment Organization proposed: "The area of ​​urban green space reaches 60m 2 / person-hour is the best living environment”, and the visual feeling is the most comfortable when the green color accounts for more than 25% of the human visual field. However, the per capita green space area in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04A01H4/00
Inventor 孙振元李正红江泽慧巨关升韩蕾刘会超杨学军
Owner INST OF FORESTRY CHINESE ACAD OF FORESTRY
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