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Culture medium formula of plant callus induction and subculture multiplication of astragalus membranaceus (fisch.) bunge and culture method of astragalus membranaceus (fisch.) bunge

A technology of callus induction and medium formulation, applied in botany equipment and methods, plant regeneration, horticultural methods, etc., can solve the problems of lack of Astragalus membranaceus plant resources, increase photosynthesis, promote rapid subculture proliferation, increase The effect of placing the number of leaves

Active Publication Date: 2015-12-23
甘肃省农业科学院生物技术研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The object of the present invention is to provide a medium formula for induction and subculture propagation of Astragalus plant callus and a culture method thereof, which can obtain a large amount of Astragalus leaf callus without browning in a relatively short period of time, through biotechnology Means to solve the problem of lack of Astragalus plant resources, and provide sufficient raw materials for the industrial extraction of active ingredients of Astragalus

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] A. Selection and sterilization of explants: Astragalus leaves were used as explants. Collect healthy and disease-free Astragalus leaves, wash them 5 times with sterile water, soak them in 75% ethanol for 30 seconds, and then put them in 0.10% HgCl 2 Soak in the solution for 12.0 minutes, then wash 3 times with sterile water.

[0047] B. Sterilization of the culture medium: the callus induction medium and the subculture proliferation medium are respectively packed in 100ml triangular culture bottles, 20.0ml per bottle, and the bottle mouth is covered with air-permeable plastic sealing film, and sterilized at a pressure of 0.10MPa 20.0 minutes, cool for later use.

[0048] C. Callus induction: take the sterilized leaves of Astragalus membranaceus and cut them into 2.0 mm long sections with a sterile scalpel, and inoculate them on the sterilized callus induction medium. The formula of callus induction medium is: MS+6-BA1.5mg / L+NAA0.8mg / L+sucrose 20.0g / L+glucose 10.0g / L+a...

Embodiment 2

[0051] A. Selection and sterilization of explants: Astragalus leaves were used as explants. Collect healthy and disease-free Astragalus leaves, wash them three times with sterile water, soak them in ethanol with a volume concentration of 72% for 40 seconds, and then put them in HgCl with a mass concentration of 0.12%. 2 Soak in the solution for 8.0 minutes, then wash 4 times with sterile water.

[0052] B. Sterilization of the culture medium: the callus induction medium and the subculture proliferation medium are respectively packed in 100ml triangular culture bottles, each bottle is 22.0ml, the bottle mouth is covered with a breathable plastic sealing film, and sterilized at a pressure of 0.15MPa 20.0 minutes, cool for later use.

[0053] C. Callus induction: take the sterilized leaves of Astragalus membranaceus and cut them into 4.0 mm long sections with a sterile scalpel, and inoculate them on the sterilized callus induction medium. The formula of callus induction medium ...

Embodiment 3

[0056] A. Selection and sterilization of explants: Astragalus leaves were used as explants. Collect healthy and disease-free Astragalus leaves, wash them three times with sterile water, soak them in 70% ethanol for 50 seconds, and then put them in 0.15% HgCl 2 Soak in the solution for 5.0 minutes, then wash 5 times with sterile water.

[0057] B. Sterilization of the culture medium: the callus induction medium and the subculture proliferation medium are respectively packed in 100ml triangular culture bottles, each bottle is 25.0ml, the bottle mouth is covered with a breathable plastic sealing film, and sterilized at a pressure of 0.12MPa 20.0 minutes, cool for later use.

[0058] C. Callus induction: take the sterilized leaves of Astragalus membranaceus and cut them into 5.0 mm long sections with a sterile scalpel, and inoculate them on the sterilized callus induction medium. The formula of callus induction medium is: MS+6-BA1.0mg / L+NAA0.6mg / L+sucrose 18.0g / L+glucose 8.0g / L+...

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Abstract

The invention relates to a culture medium formula of plant callus induction and subculture multiplication of astragalus membranaceus (fisch.) bunge and a culture method of the astragalus membranaceus (fisch.) bunge. Four steps of sterilizing selected explants, sterilizing a culture medium, inducing callus induction and carrying out callus subculture multiplication are included. A plant callus culture method of astragalus membranaceus (fisch.) bunge comprises the following steps: by taking astragalus membranaceus (fisch.) bunge leaves as the explants, adding a culture medium prepared from maltose, glucose, sucrose, agar powder, sodium alginate, naphthylacetic acid and 6-benzyl purine and the like into an MS basic culture medium to carry out tissue culture to obtain astragalus membranaceus (fisch.) bunge leaf callus; adjusting the pH value of the culture medium, and filling triangular culture bottles with a prepared callus induction culture medium and a prepared callus subculture multiplication culture medium respectively, and covering ventilated plastic sealing films on bottle openings; and sterilizing at the pressure of 0.10-0.15MPa and carrying out callus culture. According to the culture medium formula and the culture method, provided by the invention, the astragalus membranaceus (fisch.) bunge leaf calluses can be effectively induced and rapidly multiplied, and a lot of the astragalus membranaceus (fisch.) bunge leaf calluses can be obtained in relatively short time; and the astragalus membranaceus (fisch.) bunge leaf calluses have the characteristics of short production period and rapid multiplication speed and are suitable for industrial production.

Description

technical field [0001] The invention relates to a plant callus culture method, in particular to a culture medium formula for induction and subculture proliferation of Astragalus membranaceus plant callus and a culture method thereof. Background technique [0002] Astragalus (Astragalus sembranaceus (Fisch.) Bunge) is Fabaceae (Leguminosa esp.) Astragalus (AstragalusLinn.) plant, Astragalus contains saponins, sucrose, polysaccharides, various amino acids, folic acid, selenium, zinc, copper and other trace elements. Astragalus membranaceus has a high medicinal value. Raw astragalus membranaceus is beneficial for strengthening qi, diuresis, detumescence, detoxification, and muscle growth. It is suitable for spontaneous sweating, night sweats, blood numbness, edema, carbuncle and gangrene without ulceration or long-term ulceration. embolism. Honey-roasted Astragalus has the functions of invigorating qi, nourishing blood, and benefiting the middle. It is suitable for internal...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 赵瑛罗俊杰王红梅张运晖崔明九叶春雷欧巧明薛莲
Owner 甘肃省农业科学院生物技术研究所
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