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Building method of Vaccinium dunalianum suspension cell culture system

A culture system and suspension cell technology, which is applied in the field of plant cell engineering, can solve the problems that the cell suspension culture technology has not been reported, and achieve the effects of favorable proliferation, strong embryonic quality, and good quality

Active Publication Date: 2019-12-31
SOUTHWEST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, domestic scholars have used cell suspension culture technology to grow ginseng (Panax ginseng), American ginseng (P. Many studies have been carried out on the regulation of the production of secondary metabolites in medicinal plants such as Ginkgo biloba and Ginkgo biloba, but the application of cell suspension culture technology in Vaccinium camphora has not been reported yet.

Method used

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  • Building method of Vaccinium dunalianum suspension cell culture system
  • Building method of Vaccinium dunalianum suspension cell culture system
  • Building method of Vaccinium dunalianum suspension cell culture system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] (1) Processing of Vaccinium camphora leaves: cut the young tender leaves of Vaccinium camphora growing vigorously, without lignification, and moderately sized tissue culture seedlings, draw strip wounds on the front or back of the blades, and cut the leaves of the leaves The back grows close to the medium; inoculated on the WPM induction medium, the callus can grow in about two months, and the callus is translucent and highly active.

[0044] The culture conditions are: 25±2°C, light intensity controlled at 30-50 μmol / m 2 s (light conditions can choose any value within this range), the culture time is 60d; the culture medium is the combination of WPM basic medium and auxin: WPM+1.0~2.0mg / L 6-BA+ 0.05~1.0mg / L NAA+1.0~2.0 mg / L 2,4-D+30g / L sucrose+5g / L agar; (The composition of the medium is shown in Table 1).

[0045] Table 1 The effect of different hormone ratios on callus induction

[0046]

[0047] As can be seen from Table 1: the Vaccinium camphora callus can be ...

Embodiment 2

[0058] (1) Processing of Vaccinium camphora leaves: cut the young tender leaves of Vaccinium camphora growing vigorously, without lignification, and moderately sized tissue culture seedlings, draw strip wounds on the front or back of the blades, and cut the leaves of the leaves The back grows close to the medium; inoculated on the WPM induction medium, the callus can grow in about two months, and the callus is translucent and highly active.

[0059] The culture conditions are: 25±2°C, light intensity 30-50μmol / m 2 s, the culture time is 60d; the medium is the combination of WPM basic medium and auxin: WPM+2.0mg / L 6-BA+0.05mg / L NAA+2.0mg / L2,4-D+ 50g / L sucrose+5g / L agar.

[0060] (2) Proliferation of callus: select light yellow, translucent, non-browning, well-grown callus, cut into smaller pieces, if the callus is fluffy enough, gently break off the callus of suitable size Tissues, in order to reduce cut surface damage, in order to reduce late browning phenomenon; transfer to...

Embodiment 3

[0069] All the steps of this example are the same as in Example 2, except that the suspension culture cells II obtained in Example 2 are passed through a 200-mesh sieve to remove the culture medium, the tissue on the upper part of the sieve is retained, the water is blotted with sterile filter paper, and inoculated to 3 / 5 Organic modified WPM medium, continue to cultivate.

[0070] The culture medium is a combination of WPM basic medium and auxin: WPM (3 / 5 organic) + 1.0-2.0 mg / L ZT + 0.1-0.2 mg / L NAA + 28-42 g / L sucrose.

[0071] The culture conditions are: 25±2°C, dark culture, culture time 14d, medium pH value 4.8-5.2, shaker speed 150rpm / min, medium volume 50mL, inoculum volume 40g / L.

[0072] The influence of table 3 different culture medium on suspension culture system

[0073] Medium type Biomass (g) 1 / 5 Organic 0.3452 3 / 5 Organic 0.3600 ordinary 0.3271 2 times a lot 0.3532

[0074] As can be seen from Table 3, the biomass accu...

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Abstract

The invention discloses a building method of a Vaccinium dunalianum suspension cell culture system and belongs to the field of plant cell engineering. The method is characterized in that the young andtender leaf of Vaccinium dunalianum is used as the explant, the processed young and tender leaf is placed into an induction culture medium to induce a callus, transferring the callus into a proliferation culture medium to allow the callus to grow into a state suitable for suspension culture, and transferring the appropriate callus into a liquid culture medium to obtain a stable suspension culturesystem. The method has the advantages that production can be performed by selecting the corresponding culture combination according to actual production, and accordingly production cost can be lowered, efficiency can be increased, and a guidance basis is provided for the industrialized production of the Vaccinium dunalianum suspension culture system.

Description

technical field [0001] The invention relates to a method for establishing a suspension cell culture system of bilberry camphor leaves, belonging to the field of plant cell engineering. Background technique [0002] Vaccinium dunalianum (Vaccinium dunalianum) is an evergreen shrub of the genus Vaccinium in the Ericaceae family, and is mainly distributed in the northwestern part of Yunnan through the Central Yunnan Plateau to southern and eastern Yunnan. Its fruit is rich in nutrition, and the whole plant has the effects of relaxing tendons and activating collaterals, expelling wind and dehumidification, and has high medicinal and health value. In addition, Vaccinium camphora is also a special resource plant rich in caffeoyl arbutin. It is developed and utilized as a natural alternative resource of arbutin. Vaccinium camphor is still in the wild at present, and its wild resources have been drastically reduced due to destructive harvesting year after year. Therefore, it is of...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005A01H4/008
Inventor 赵平唐军荣刘云阚欢付羚罗旭璐张訸
Owner SOUTHWEST FORESTRY UNIVERSITY
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