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Regeneration method of Camellia oleifera regeneration system

A system and technology of Camellia oleifera, applied in the directions of plant regeneration, horticultural methods, botanical equipment and methods, etc., can solve the problems of low callus induction rate, low differentiation rate, can not meet the requirements of factory seedling raising, etc., to improve the induction rate and Differentiation rate, the effect of improving the differentiation rate

Active Publication Date: 2019-12-24
RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Abstract
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  • Claims
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Problems solved by technology

[0006] But at present, the tissue culture of camellia oleifera is mainly to carry out the tissue culture of the stem section with axillary buds. This method is easy to carry out tissue culture and rapid propagation, but there is a problem of high pollution rate in the stem section culture, and it cannot be regenerated; Rapid growth of seedlings by tissue culture, but this method cannot maintain the traits of good varieties; there are also studies on the use of leaves for tissue culture regeneration, but the induction rate of leaf callus is low, and the differentiation rate is very low, which cannot meet the requirements of industrial seedlings

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  • Regeneration method of Camellia oleifera regeneration system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Present embodiment provides a kind of regeneration method of camellia oleifera sheet, as figure 1 shown, including:

[0033] (1) Disinfection of explants

[0034] Pick the young leaves of common Camellia oleifera 'Changlin No. 4', soak in detergent for 5-10 minutes, then rinse with tap water for 1-2 hours, and dry the water with absorbent paper. On the ultra-clean workbench, soak the leaves with 70%-75% alcohol for 10-30s, wash them with sterile water for 3-4 times, then disinfect the surface with 0.08-0.12% HgCl2 for 8-12min, and then wash them with sterile water Wash 6-8 times.

[0035] (2) Callus induction culture

[0036] The specific cultivation includes: putting the leaves on sterile filter paper, drawing 2-4 lines perpendicular to the main vein in the middle of the leaves with a scalpel, and then peeling off the leaves with tweezers to inoculate the leaves on the callus induction medium. 3-8 leaves were inoculated per dish (90 mm). Place the inoculated cultu...

Embodiment 2

[0050] The present embodiment provides a kind of regeneration method of camellia oleifera sheet, comprising:

[0051] (1) Callus induction culture

[0052] The leaf material is the leaf of the aseptic tissue culture seedling of 'Changlin No. 4' common Camellia oleifera, and the specific cultivation is as in Example 1.

[0053] The callus induction medium is: 1 / 2MS basic medium (powder), 1.0mg / L of 2,4-D, 0.5mg / L of TDZ, 50g / L of sucrose, 8g / L of agar, 250mg / L hydrolyzed casein, 250mg / L hydrolyzed milk protein, 100mg / L proline, 100mg / L glycine, 100mg / L glutamine, 100mL / L coconut water, and 1g / L PPVP and 10mg / L silver nitrate, and adjust the pH to 5.6; the callus induction rate is 100%.

[0054] (2) Callus differentiation culture

[0055] The specific culture is as in Example 1.

[0056] The differentiation medium is: 1 / 2MS medium (powder), 0.05mg / L NAA, 1.0mg / L 6-BA, 0.05mg / L TDZ, 30g / L sucrose, 7g / L agar , 300mg / L hydrolyzed casein, 300mg / L hydrolyzed milk protein, 150mg...

Embodiment 3

[0062] The present embodiment provides a kind of regeneration method of camellia oleifera sheet, comprising:

[0063] (1) Callus induction culture

[0064] The leaf material is the leaf of the aseptic tissue culture seedling of 'Changlin No. 4' common Camellia oleifera, and the specific cultivation is as in Example 1.

[0065] The callus induction medium is: 1 / 2MS basic medium (powder), 1.0mg / L of 2,4-D, 1.0mg / L of TDZ, 70g / L of sucrose, 8g / L of agar, 500mg / L hydrolyzed casein, 500mg / L hydrolyzed milk protein, 150mg / L proline, 150mg / L glycine, 150mg / L glutamine, 100mL / L coconut water, and 2g / L PPVP and 10mg / L silver nitrate, and adjust the pH to 5.8; the callus induction rate is 100%.

[0066] (2) Callus differentiation culture

[0067] The specific culture is as in Example 1.

[0068] The differentiation medium is: 1 / 2MS medium (powder), 0.05mg / L NAA, 2.0mg / L 6-BA, 0.01mg / L TDZ, 300g / L sucrose, 7g / L agar , 500mg / L hydrolyzed casein, 500mg / L hydrolyzed milk protein, 200m...

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Abstract

The invention belongs to the technical field of plant tissue regeneration and particularly relates to a regeneration method of a Camellia oleifera regeneration system. The method includes: callus induction culture, callus differentiation culture, rooting culture and seedling acclimation, wherein the callus induction culture adopts dark culture, and a callus induction culture medium used by the callus induction culture comprises a hormone component, an anti-browning agent and sucrose with the concentration being 30-80g / L. The method has the advantages that the callus induction culture adoptingdark culture is used, the specific hormone component and the anti-browning agent are added into the callus induction culture medium, the sucrose concentration is increased, the multiple technical means generate a synergistic effect, and accordingly the callus induction rate and differentiation rate of Camellia oleifera leaf callus can be increased evidently; by the method, the efficiency Camelliaoleifera seedling culture using tissue culture can be increased evidently, and a foundation can be laid for the building of a Camellia oleifera genetic transformation system.

Description

technical field [0001] The invention belongs to the technical field of plant tissue regeneration, and in particular relates to a regeneration method of camellia oleifera regeneration system. Background technique [0002] Camellia oleifera (Camellia oleifera) is an evergreen small tree or shrub of the genus Camellia in the family Theaceae (Theaceae). . [0003] Camellia oil contains about 90% unsaturated fatty acids, rich in vitamin E, vitamin D, vitamin K and vitamin A, and also contains trace amounts of squalene, flavonoids, tea polyphenols and camellia glycosides. Regular consumption of camellia oil can prevent cardiovascular sclerosis, lower blood pressure, lower blood fat, etc., so camellia oil is recognized as an ideal nutritional and health-care edible oil. [0004] Camellia oil can be used as lubricating oil and antirust oil in industry, can be used to produce oleic acid and its esters, produce soap and vaseline, etc.; in medicine, it can be used to make injections ...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 叶思诚曹永庆姚小华卓仁英王开良张燕
Owner RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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