Aralia elate seem hormone autotrophic cell line

A kind of Aralia aralia, self-supporting technology, applied to plant cells, fermentation, etc.

Inactive Publication Date: 2010-01-20
NORTHEAST FORESTRY UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Aralia elate seem hormone autotrophic cell line
  • Aralia elate seem hormone autotrophic cell line
  • Aralia elate seem hormone autotrophic cell line

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0016] Example 1 The acquisition of Aralia longya hormone autotrophic cell line

[0017] 1) Source of material

[0018] The seeds of Aralia longya were collected from the seed garden of Heilongjiang Forestry Diversification Research Institute (Mudanjiang City) in early October 2007.

[0019] 2) Stratification treatment of Aralia japonica seeds to break dormancy

[0020] Since the zygotic embryos in the mature fruits of Aralia japonica have not matured, there is dormancy in seed germination. To break the dormancy, post-ripening treatment of zygotic embryos is necessary. The processing method we use is layered processing. That is to say, the seeds are mixed with wet sand in proportion after simple disinfection treatment, and placed in a refrigerator at 4°C for stratification treatment.

[0021] 3) Cultivation of sterile seedlings

[0022] After 4 months of seed stratification, select the pod-opened seeds with mature zygotic embryos, take out the zygotic embryos after routine...

example 2

[0025] The growth amount determination of example 2 hormone autotrophic cell lines

[0026] Cells with a fresh weight of 0.16-0.18g were placed in four 100ml glass Erlenmeyer flasks filled with 30ml 1 / 2MS solid medium and cultured for 3 weeks. The growth of the four flasks was 7.81g fresh weight and 0.48g dry weight. The weight increased by 11 times, and the fresh weight increased by 10.8 times.

[0027] The growth situation of table 1 hormone autotrophic cell line cultured in solid 1 / 2MS medium for 3 weeks

[0028]

example 3

[0029] Determination of Example 3 Hormone Autotrophic Cell Line Total Saponin Content

[0030] 1) Preparation of the test solution

[0031] Take 0.5 g of dried Aralia japonica powder, weigh it accurately, add 15 mL of absolute ethanol solution, and oscillate ultrasonically for 25 min. Afterwards, extract in a constant temperature water bath at 70°C for 3 times, 1 hour each time, collect and combine extracts, and evaporate to dryness. The residue was dissolved in 5 mL of water and extracted with ether until the ether layer was colorless. The combined aqueous solution was collected and evaporated to dryness. The residue was dissolved in methanol and the volume was adjusted to 50ml to obtain a sample solution of total saponins of Aralia japonica.

[0032] 2) Drawing of standard curve

[0033] Oleanolic acid standard was purchased from China Institute of Pharmaceutical and Biological Products, batch number (110709-200505). Accurately weigh 5 mg of oleanolic acid standard subst...

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Abstract

The invention belongs to the plant biotechnology field. Leaves and roots of seedlings of an Aralia elate seem are taken as an explant, and a loose yellow hormone autotrophic cell line Aral-1 in fast growth is induced from a culture medium of MS+2,4-D (0.1-5.0mg/L) plus sucrose (2%). Fresh weight and dry weight of the cell line are increased by about 11 times after three-week growth in a 1/2 MS culture medium without exo-hormones; and contents of the total saponin and the total flavone are respectively 7.10% and 1.51% measured by an ultraviolet spectrophotometer, wherein, content of the total saponin is obviously higher than that of a white hormone-dependant callus tissue and is 60% of that of a reference wild leafstalk, with a little difference from that of a reference wild leafstalk; and content of the total flavone is obviously higher than that of the white hormone-dependant callus tissue and the wild leafstalk, and is 68.9% of that of the wild leafstalk. The cell line has the characteristics of fast growth without hormones, good dispersion phase in liquid, relatively higher pharmaceutical ingredient content and the like, and can be used as a material for producing pharmaceutical ingredients by mass cell culture.

Description

technical field [0001] The invention belongs to the field of biotechnology, and mainly relates to a hormone autotrophic Aralia japonica cell line obtained through a tissue culture method with loose texture, yellow color and rapid growth. Compared with wild leaves, this cell line contains higher medicinal secondary metabolites flavonoids and aralia saponins, and can be used for large-scale cultivation of Aralia japonica cells to produce medicinal ingredients. Background technique [0002] Aralia elate Seem., a small tree belonging to the Araliaceae genus Aralia, is a plant with both food and medicine. Its sprouts contain various amino acids and trace elements necessary for the human body, and are a good health food. The main medicinal components of Aralia japonica are aralia saponins and flavonoids. The main constituent of its saponins is oleanolic acid. Oleanolic acid has anti-inflammatory, sedative, diuretic, cardiotonic, immune and anti-cancer effects, especially for th...

Claims

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Application Information

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IPC IPC(8): C12N5/04C12P19/56C12P17/06
Inventor 曲冠证由香玲詹亚光代金玲谭啸崔龙义李在善
Owner NORTHEAST FORESTRY UNIVERSITY
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