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Mirabilis himalaica callus, proliferation method and suspension cell reproduction method

A technology of callus and callus induction, applied in plant cells, tissue culture, cell culture medium, etc., can solve the problems of being easily affected by environmental conditions, long growth cycle, and difficult to meet the large demand of Mirabilis Himalayan, Achieve the effect of improving reproduction effect, short growth cycle and low cost

Active Publication Date: 2019-11-01
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Although great progress has been made in artificial planting of Mirabilis Himalayan in recent years, it is difficult to meet the large demand for Mirabilis Himalayan in the industrialization of Tibetan medicine because the medicinal parts of Mirabilis Himalayan are perennial roots with a long growth cycle and are easily affected by environmental conditions.

Method used

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  • Mirabilis himalaica callus, proliferation method and suspension cell reproduction method
  • Mirabilis himalaica callus, proliferation method and suspension cell reproduction method
  • Mirabilis himalaica callus, proliferation method and suspension cell reproduction method

Examples

Experimental program
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Effect test

Embodiment 1

[0056] (1) Cultivation of sterile vaccines

[0057] Take Himalayan Mirabilis (for plant morphology see figure 1 ) Mature seeds are disinfected according to the conventional disinfection method of 75% alcohol+2% sodium hypochlorite combination, inoculated on MS medium, and cultivate aseptic seedlings. Culture conditions: 3000 lx light, 16 hours of light and 8 hours of darkness per day, set temperature at 25°C, and culture time of 25 days.

[0058] (2) Induction of callus

[0059] The leaves of the aseptic seedlings (seedling leaves) cultivated in step (1) are cut into 0.5 cm×0.5 cm, inserted into induction medium for callus induction culture, and primary callus is obtained. The medium formula is: MS+1.5mg / L TDZ+2.0mg / LNAA. Energy substance: sucrose 30g / L; support: agar 6.5g / L; pH: pH5.8; solvent: deionized water in the composition of the induction medium; culture conditions: dark, set temperature 25°C; culture time :35 days.

[0060] After 10 days of explant inoculation, t...

Embodiment 2

[0066] Carry out callus and suspension cell culture to Mirabilis Himalayan in the same manner as Example 1, the difference is:

[0067] The culture condition of aseptic seedling is light 2500lx, and culture time is 30 days.

[0068] The formula of callus induction medium is: MS+3.0mg / L TDZ+4.0mg / L NAA, energy substance: sucrose 25g / L; support: agar 7.0g / L; culture time: 30 days.

[0069] Subculture callus proliferation medium formula: MS+0.5mg / L TDZ+3.0mg / L NAA, energy substance: sucrose 25g / L; support: agar 7.0g / L; culture time: 20 days, cycle 15 Second-rate.

[0070] Suspension cell medium formula is: MS+2.0mg / L TDZ+3.0mg / L NAA+300mg / L hydrolyzed casein, energy substance: sucrose 25g / L; culture conditions: shaker speed 120r / min, culture time: 15 days, cycle 10 times.

Embodiment 3

[0072] Carry out callus and suspension cell culture to Mirabilis Himalayan in the same manner as Example 1, the difference is:

[0073] The culture condition of aseptic seedling is light 3500lx, and culture time is 20 days.

[0074] The formula of callus induction medium is: MS+0.5mg / L TDZ+2.0mg / L NAA, culture time: 40 days.

[0075] The formula of subculture callus proliferation medium is: MS+2.0mg / L TDZ+3.0mg / L NAA, culture time: 30 days, cycle 20 times.

[0076] Suspension cell medium formula is: MS+0.5mg / L TDZ+1.5mg / L NAA+100mg / L hydrolyzed casein, culture time: 30 days, cycle 5 times.

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Abstract

The invention relates to the technical field of plant tissue culture and particularly discloses mirabilis himalaica callus, proliferation method and suspension cell reproduction method. In the mirabilis himalaica callus induced proliferation method, an induction medium carries out induction culture on blades of aseptic seedlings of mirabilis himalaica to form callus, and a proliferation medium performs further proliferation culture on the callus to obtain subculture callus; wherein hormones in the induction and proliferation media are TDZ and NAA. The method is high in inductivity, little in differentiation and short in growth period, the obtained callus is soft and fragile, and follow-up utilization is facilitated. The invention also provides a mirabilis himalaica suspension cell reproduction method for suspension culture of the prepared mirabilis himalaica callus by a liquid medium containing the TDZ, the NAA and casein hydrolysate. The method is simple to operate, high in cell proliferation rate, short in growth period, low in cost and capable of realizing industrial year-round production.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture. Specifically, it relates to a callus of Mirabilis jasmine Himalayan, a propagation method and a suspension cell propagation method. Background technique [0002] The Qinghai-Tibet Plateau is the largest plateau in China and the highest in the world, known as the "roof of the world" and the "third pole" of the earth. It has strong radiation, more sunshine, low temperature, less accumulated temperature, the temperature decreases with the increase of altitude and latitude, and the temperature difference between day and night is large. and other typical plateau climate characteristics. Under such environmental conditions, only those plants that have acquired special ecological-biological adaptation characteristics in the tenacious struggle against harsh climatic conditions such as strong radiation, ice and snow, severe cold, strong wind, flowstone, and barren for generations can surviv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04C12N5/02
CPCC12N5/0025C12N5/04
Inventor 陈玉珍卢存福
Owner BEIJING FORESTRY UNIVERSITY
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