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Callus culture method of snow lotus herb

A technology of callus and culture method, which is applied in the field of Tianshan Snow Lotus callus culture, can solve the problems of slow growth of wild Tianshan Snow Lotus, endangered wild Tianshan Snow Lotus, low seed reproduction rate, etc., and achieves extremely market competitiveness, cycle cycle Short, non-polluting effect on the environment

Inactive Publication Date: 2010-07-14
烟台汇鹏生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to its special growth environment, wild Tianshan snow lotus grows slowly, and it takes 5 to 6 years from seed germination to flowering and fruiting, and the reproductive rate of seeds is very low, wild resources are severely limited, and wanton illegal harvesting makes Wild Tianshan snow lotus endangered
So far, no abiotic elicitors have been systematically studied on the regulation of the biosynthesis of sweet components of Lilac in Saussurea tianshanensis culture

Method used

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  • Callus culture method of snow lotus herb
  • Callus culture method of snow lotus herb

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: a kind of Tianshan Snow Lotus callus culture method, it comprises the steps:

[0027] a. Plant selection and aseptic treatment:

[0028] a) Select the young stems and leaves of wild Tianshan snow lotus fresh plants with an altitude of more than 3500m in July to wash the soil with water, and then soak in distilled water; b) soak the cleaned young stems and leaves with 75% alcohol for 15 seconds to carry out surface sterilization c) Soak the young stems and leaves sterilized by alcohol in 0.1% mercuric chloride solution for 5 minutes, and constantly stir gently during the soaking process so that the surface of the plant body fully contacts the mercuric chloride solution; The mercuric chloride solution soaked and sterilized plants were rinsed 5 times with high-temperature sterilized distilled water, and the mercuric chloride solution adhering to the surface of the plants was rinsed clean, and then the plants were blotted dry with sterile filter paper; Bacter...

Embodiment 2

[0033] Embodiment 2: a kind of Tianshan Snow Lotus callus culture method, it comprises the steps:

[0034] a. Plant selection and aseptic treatment:

[0035] a) Select the young stems and leaves of wild Tianshan snow lotus fresh plants with an altitude of more than 3800m in August to wash the soil with water, and then soak in distilled water; b) soak the cleaned young stems and leaves with 78% alcohol for 12 seconds to carry out surface sterilization c) Soak the young stems and leaves sterilized by alcohol in 0.12% mercuric chloride solution for 4 minutes, and constantly stir gently during the soaking process to make the plant surface fully contact the mercuric chloride solution; The mercuric chloride solution soaked and sterilized plants were rinsed 5 times with high-temperature sterilized distilled water, and the mercuric chloride solution adhering to the surface of the plants was rinsed clean, and then the plants were blotted dry with sterile filter paper; Bacteria scissor...

Embodiment 3

[0040] Example 3: The wild Tianshan Snow Lotus, the callus cultured in Example 1 and Example 2 were detected.

[0041] Use HPLC chromatographic detection method in the chromatographic column is Diamonsil ODS C-18 alkyl silica gel column (4.6mm * 250mm); Mobile phase: methanol-water (20: 80); UV detection wavelength: 262nm; Column temperature: 40 ℃; Sample volume: 10 μl; flow rate: 1.0ml / min to detect the content of lilacin.

[0042] a) Preparation of syringin standard solution: Accurately weigh 2.9 mg of syringin standard substance into a 10 ml volumetric flask, dissolve in mobile phase, and prepare 290 μg·mL-1 reference substance stock solution.

[0043] b) Drawing of syringin standard curve: draw 0.05, 0.20, 0.40, 0.60, 0.80, 1.00, 1.50, 2.00ml of the above stock solution respectively and place them in a 5ml volumetric flask, dilute to the mark with mobile phase, and take 5μl of each dilution Injection. Linear regression was performed with peak area (A) versus syringin con...

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Abstract

The invention discloses a callus culture method of snow lotus herb, comprising the steps of selection and asepsis of plants, induction and culture of callus and large-scale culture. The core technology of the method is selection of the following large-scale culture medium: MS+0-50g / L of cane sugar+0-10g / L of agar+0.1-5.0mg / L of alpha-naphthylacetic acid+0.1-5.0mg / L of 6-benzyladenine. During large-scale culture, an abiotic elicitor, 40-60mu mol / L of SNP+40-60mu mol / L of MJ+40-60mu mol / L of SA is added. The method is easy to operate, and the active ingredient is high in content and low in production cost, does not pollute environment and can reach the level of industrialization. Syringin produced by the culture technology in the invention is invariant, high in content, low in cost and short in cycle and is very competitive in the markets.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for culturing callus of Snow Lotus in Tianshan Mountains. Background technique [0002] Tianshan Snow Lotus (Saussurea involucrate Kar.et Kir.) belongs to Angiosperm, Dicotyledoneae, Compositae, Asteraceae, Artichoke family, and Phyllostachys genus. As early as the eighth century, there were records of snow lotus in the ancient Tibetan medical literature "Yuewang Yaozheng", and later in bibliographies such as "Northwest Territory Records", "Jingzhu Materia Medica", "Lan Liuli", "Compendium of Materia Medica Supplements" and other bibliographies. recorded. It is mainly distributed in the alpine regions such as Xinjiang, Qinghai, Gansu, Yunnan and Tibet in my country, between rock crevices and cliffs near the snow line at an altitude of 2400-4100m, and rarely distributed abroad. Snow lotus is warm in nature and slightly bitter in taste. It enters...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 贾景明
Owner 烟台汇鹏生物科技有限公司
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