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A kind of medium formula and method for preventing tissue culture browning of Magnolia japonica

A cultivation method and technology of Magnolia serrata are applied in horticultural methods, botanical equipment and methods, horticulture and other directions, and can solve the problem of the formula and method of tissue culture browning medium without buds of Magnolia serrata, and the effect of preventing browning from being quite different. The results are unsatisfactory and other problems, and the effect of preventing browning is remarkable, reducing the probability of bacterial contamination, and inhibiting the strong activity of polyphenol oxidase.

Inactive Publication Date: 2017-10-31
BEIHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although it has a certain effect of preventing browning, for tissue culture of buds taken in different months, the effect of preventing browning is quite different, and the result is still unsatisfactory. The formula and method of the base

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] (1) Material collection: Take the annual branches in early April, and cultivate them indoors for 8-10 days. After the buds germinate, take the terminal buds and side buds (0.5-1.0 cm in length), soak them in detergent solution for 20 minutes, and rinse them with running water. 2h, then disinfect the surface with 75% ethanol on the ultra-clean workbench for 30s, rinse with sterile water for 2-3 times, and then wash with 0.1% HgCl 2 Treat for 7-8 minutes, rinse with sterile water 5-6 times, and inoculate.

[0018] (2) Acquisition of sterile seedlings (primary culture): inoculate the sprouts sterilized in step (1) in the culture medium, add 0.5mg / L 6-BA, 0.3mg / L NAA, Vc 300mg / L, GSH 100mg / L, sodium benzoate 30mg / L and sucrose 30g / L, pH5.6~5.8. The light time is 12 hours, the culture temperature is 23±2°C during the day, and the light intensity is 1500-2000lx at 20±2°C at night. After 30 days of cultivation, sterile seedlings are obtained, and the browning rate is less tha...

Embodiment 2

[0023] (1) Material collection: Take the branches of the current year in mid-May, cut the terminal buds and lateral buds (length 0.5-1.0cm) in the laboratory, soak them in the detergent solution for 20 minutes, rinse them with running water for 2 hours, and then work in the ultra-clean room. Disinfect the surface with 75% ethanol on the stage for 30 seconds, rinse it with sterile water for 2-3 times, and then wash it with 0.1% HgCl 2 Treat for 7-8 minutes, rinse with sterile water 5-6 times, and inoculate.

[0024] (2) Acquisition of sterile seedlings (primary culture): inoculate the sprouts sterilized in step (1) in the culture medium, add 0.5mg / L 6-BA, 0.3mg / L NAA, Vitamin C 300mg / L, GSH 100mg / L, sodium benzoate 50mg / L and sucrose 30g / L, pH5.6-5.8. The light time is 12 hours, the culture temperature is 23±2°C during the day, and the light intensity is 1500~2000lx at 20±2°C at night. After 30 days of cultivation, sterile seedlings are obtained, and the browning rate is less ...

Embodiment 3

[0029](1) Material collection: Take the branches of the current year in mid-June, cut the terminal buds and side buds (length 0.5-1.0cm) in the laboratory, soak them in the detergent solution for 20 minutes, rinse them with running water for 2 hours, and then work in the ultra-clean room. Disinfect the surface with 75% ethanol on the stage for 30 seconds, rinse it with sterile water for 2-3 times, and then wash it with 0.1% HgCl 2 Treat for 7-8 minutes, rinse with sterile water 5-6 times, and inoculate.

[0030] (2) Acquisition of sterile seedlings (primary culture): inoculate the sprouts sterilized in step (1) in the culture medium, add 0.5mg / L 6-BA, 0.3mg / L NAA, Vitamin C 400mg / L, GSH 100mg / L, sodium benzoate 50mg / L and sucrose 30g / L, pH5.6-5.8. The light time is 12 hours, the culture temperature is 23±2°C during the day, and the light intensity is 1500~2000lx at 20±2°C at night. After 30 days of cultivation, sterile seedlings are obtained, and the browning rate is less tha...

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Abstract

The invention relates to a medium composition for preventing magnolia sieboldii bud tissue culture browning and a method. The method comprises the five steps of explant sterilization, medium obtaining through aseptic seedlings, medium subculture and medium rooting and transplanting. According to the culture method, buds are adopted as an explant, cytokinin, auxin, vitamin C, glutathione, sodium benzoate, saccharose and agar are added into a B5 culture medium to form the medium composition for preventing browning, and prevention and control over browning in the magnolia sieboldii bud tissue culture process are achieved. The method is good in repeatability, the bud browning rate is lower than 10%, the rooting rate is 75-80%, the transplanting survival rate is 85-90%, and the method is of great significance in magnolia sieboldii seed quality resource protection and development and utilization thereof.

Description

technical field [0001] The invention belongs to the field of tissue culture, and relates to a medium formula and a method for preventing tissue culture browning of magnolia magnolia buds. Background technique [0002] Heavenly Maiden Mulan (Magnolia sieboldii K. Koch) is the magnoliaceae (Magnoliaceae) Magnolia (Magnolia) deciduous small trees, the national third-class endangered tree species. Tiannv Magnolia has both ornamental, aromatic and medicinal values, and is a rare wild woody plant resource with great development prospects. [0003] In the process of tissue culture of Magnolia tiannv buds, browning of explants often occurs. Browning seriously affects the dedifferentiation and redifferentiation of explants, resulting in culture failure. This has become a thorny problem in the tissue culture of Magnolia chinensis, and it has also become a major obstacle to the establishment of the rapid propagation system of Magnolia chinensis. It is not yet satisfactory. , the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/001
Inventor 高红兵王欢姜莹杜凤国
Owner BEIHUA UNIV