Tissue culture method of Glossostigma elatinoides

A technology of tissue culture and dwarf pearls, applied in horticultural methods, botany equipment and methods, plant regeneration, etc., can solve problems such as quality and yield decline, planting benefit decline, and yield reduction, so as to improve yield and quality, and increase load capacity. Cultivate the survival rate and realize the effect of factoryization

Active Publication Date: 2017-02-22

浙江中日弹簧有限公司

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AI-Extracted Technical Summary

Problems solved by technology

However, the traditional seedling breeding method of dwarf pearls has reduced yield, serious virus ...

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Abstract

A tissue culture method of Glossostigma elatinoides comprises the steps of acquiring and disinfecting an explant, cutting the explant into a stem with buds, inoculating to an initial medium, culturing to obtain adventitious buds, applying an ultrasonic field for 1-2 hours each during 1st to 7th days of initial culture, applying an ultrasonic field for 4-5 hours each day during 8th to 15th days of initial culture, transferring the adventitious buds to a subculture medium, culturing to obtain subculture seedlings, exposing to light for 10-12 hours per day under the temperature of 24+/-2 DEG C and the humidity of 70-75% and under the light intensity of 2800-3200 lx, transferring the subculture seedlings to a rooting medium, culturing to obtain seedlings of Glossostigma elatinoides, and finally exercising and transplanting the seedlings; the rooting medium is made by adding ammonium molybdate, TDZ and amylopectin to N6 medium; the method is insusceptible to season, temperature and territory, mass aseptic seedlings are acquired, culture success rate of seedlings is increased, and the yield and quality are increased.

Application Domain

Plant tissue cultureHorticulture methods

Technology Topic

BudAmmonium molybdate +6

Examples

Experimental program(2)

Example Embodiment

[0025] Example 1

[0026] The tissue culture method of Dwarf Pearl includes the following steps:

[0027] (1) Obtain and disinfect the explant

[0028] Choose new shoots of robust plants, wash the dirt and dust on the surface of the shoots with tap water, first soak in alcohol, then disinfect with mercury, and finally rinse with sterile water to obtain disinfected explants.

[0029] (2) Primary training

[0030] Divide the sterilized explants into shoots with buds, then put them into the primary culture medium and cultivate them for 32 days to obtain adventitious buds.

[0031] The initial culture conditions are as follows: temperature is 22℃, humidity is 63%, illumination time is 8 hours/day, light intensity is 22001x, first to 7 days of primary culture, ultrasonic field is applied for 1 hour every day, ultrasonic field power is 8W/cm 2 , The ultrasonic field frequency is 20KHz, and the ultrasonic field is applied for 4 hours every day on the 8th to 15th day of the primary culture, and the ultrasonic field power is 15W/cm 2 , The ultrasonic field frequency is 25KHz;

[0032] The primary culture medium is: on the basis of ER medium, add potassium iodate 0.1mg/L, add 2,4-D 0.6mg/L, add paclobutrazol 1.2mg/L;

[0033] (3) Subculture

[0034] The adventitious shoots are transferred to the subculture medium and cultivated for 40 to 45 days to obtain subculture seedlings.

[0035] The subculture conditions are: temperature is 26℃, humidity is 70%, light time is 10 hours/day, light intensity is 28001x,

[0036] The secondary culture medium is: on the basis of MS medium, first adjust the concentration of ammonium nitrate to 1200mg/L, then add urea 100mg/L, add ABA 3mg/L, add indole acetic acid 0.5mg/L, add glutamine Amide 5.0mg/L, add activated carbon 200mg/L;

[0037] (4) Rooting culture

[0038] Transfer the subculture seedlings to rooting medium and cultivate them for 20-25 days to obtain dwarf pearl seedlings.

[0039] The rooting culture conditions are: temperature is 28℃, humidity is 40%, light time is 12 hours/day, light intensity is 35001x,

[0040] The rooting medium is: in N 6 On the basis of the medium, add ammonium molybdate 0.01mg/L, TDZ 1.5mg/L, pullulan 130mg/L, and adjust the pH to 7.0 with sodium hydroxide;

[0041] (5) Seedling refinement and transplanting

[0042] After the rooting culture is completed, open the cap of the culture bottle, open the bottle and cultivate seedlings under natural light for 5-7 days, then rinse the dwarf pearl seedlings with water several times to remove the residual medium, and transplant them into natural water.

Example Embodiment

[0043] Example 2

[0044] The tissue culture method of Dwarf Pearl includes the following steps:

[0045] (1) Obtain and disinfect the explant

[0046] Choose new shoots of robust plants, wash the dirt and dust on the surface of the shoots with tap water, first soak in alcohol, then disinfect with mercury, and finally rinse with sterile water to obtain disinfected explants.

[0047] (2) Primary training

[0048] Divide the sterilized explants into shoots with buds, then put them into the primary culture medium and cultivate them for 36 days to obtain adventitious buds.

[0049] The initial culture conditions are as follows: temperature is 26℃, humidity is 68%, light time is 10 hours/day, light intensity is 25001x, first to 7 days of primary culture, ultrasonic field is applied for 2 hours every day, ultrasonic field power is 12W/cm 2 , The ultrasonic field frequency is 22KHz, and the ultrasonic field is applied for 5 hours every day on the 8th to 15th day of the primary culture, and the ultrasonic field power is 18W/cm 2 , The ultrasonic field frequency is 26KHz;

[0050] The primary culture medium is: on the basis of ER medium, add potassium iodate 0.5mg/L, add 2,4-D 0.8mg/L, add paclobutrazol 1.3mg/L;

[0051] (3) Subculture

[0052] The adventitious shoots are transferred to the subculture medium and cultivated for 45 days to obtain subculture shoots.

[0053] The subculture conditions are: temperature of 26℃, humidity of 75%, light time of 12 hours/day, light intensity of 32001x,

[0054] The secondary culture medium is: on the basis of MS medium, first adjust the concentration of ammonium nitrate to 1300mg/L, then add 120mg/L urea, add ABA3.5mg/L, add indole acetic acid 0.8mg/L, add Glutamine 5.5mg/L, 300mg/L activated carbon added;

[0055] (4) Rooting culture

[0056] Transfer the subculture seedlings to rooting medium and cultivate them for 20-25 days to obtain dwarf pearl seedlings.

[0057] The rooting culture conditions are: temperature is 32℃, humidity is 50%, light time is 15 hours/day, light intensity is 42001x,

[0058] The rooting medium is: in N 6 On the basis of the medium, add ammonium molybdate 0.02mg/L, add TDZ 2.0mg/L, add pullulan 150mg/L, and adjust the pH to 7.2 with sodium hydroxide;

[0059] (5) Seedling refinement and transplanting

[0060] After the rooting culture is completed, open the cap of the culture bottle, open the bottle and cultivate seedlings under natural light for 5-7 days, then rinse the dwarf pearl seedlings with water several times to remove the residual medium, and transplant them into natural water.

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Description & Claims & Application Information

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Classification and recommendation of technical efficacy words

High reproductive coefficient
Shorten the breeding cycle

Tissue culture method of Glossostigma elatinoides

AI-Extracted Technical Summary

Problems solved by technology

Abstract

Examples

Example Embodiment

Example Embodiment

PUM

Description & Claims & Application Information

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