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Fast propagation method for arachniodes exilis callus tissues and suspension cells

A technology of callus and compound leaf fern, applied in the field of plants, can solve the problems of no large-scale promotion and planting, no research on reproduction and the like

Inactive Publication Date: 2015-01-07
NANJING TONGZE AGRI SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Auricularia compound is also a kind of traditional Chinese medicine, which is used for clearing away heat, detoxifying, and treating sores. At present, its use is mainly collected in the wild, and there is no large-scale promotion and planting. It is propagated by tissue culture technology, and no one has studied it.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] Take the leaves of Auricularia compound leaf, soak them in bleaching powder for 2 minutes, remove surface stains with a brush, wash them with running water for 20 minutes, treat them with sodium hypochlorite on a clean bench for 10 minutes, rinse them with sterile water 5-7 times, and sterilize the compound leaves The formula of the culture medium for inserting fern leaves is Nitsch+TDZ0.2mg / L+IBA1mg / l+10% coconut milk induction medium for callus induction culture, additional sucrose 30g / L, agar 6.5g / L, pH5- 6. Light 1000lx, temperature 15°C, inoculate the induced callus into the medium Nitsch+2,4-D0.1mg / L+TDZ0.1mg / L+EDTA+ defoaming agent, each bottle inoculated 70ml Solution, after inoculation, placed in rotary agitation and aerated culture, added 30g / L sucrose, pH 5.8, light 1500lx, temperature 15°C, the cells cultured in suspension were placed on a solid plate, and cell lines with high content of secondary metabolites were screened.

Embodiment 2

[0011] Take the leaves of Auricularia compound leaf, soak them in bleaching powder for 2 minutes, remove surface stains with a brush, wash them with running water for 20 minutes, treat them with sodium hypochlorite on a clean bench for 10 minutes, rinse them with sterile water 5-7 times, and sterilize the compound leaves The formula of the medium for inserting fern leaves is Nitsch+TDZ0.2mg / L+IBA3mg / l+10% coconut milk induction medium for callus induction culture, adding sucrose 30g / L, agar 6.5g / L, pH5- 6. Light 1000lx, temperature 15°C, inoculate the induced callus into the medium Nitsch+2,4-D0.2mg / L+TDZ0.2mg / L+EDTA+defoamer in the Erlenmeyer flask, inoculate 70ml in each bottle Solution, after inoculation, placed in rotary agitation and aerated culture, added 30g / L sucrose, pH 5.8, light 1500lx, temperature 15°C, the cells cultured in suspension were placed on a solid plate, and cell lines with high content of secondary metabolites were screened.

Embodiment 3

[0013] Take the leaves of Auricularia compound leaf, soak them in bleaching powder for 2 minutes, remove surface stains with a brush, wash them with running water for 20 minutes, treat them with sodium hypochlorite on a clean bench for 10 minutes, rinse them with sterile water 5-7 times, and sterilize the compound leaves The formula of the medium for inserting fern leaves is Nitsch+TDZ0.2mg / L+IBA2mg / l+10% coconut milk induction medium for callus induction culture, adding sucrose 30g / L, agar 6.5g / L, pH5- 6. Light 1000lx, temperature 15°C, inoculate the induced callus into the medium Nitsch+2,4-D0.2mg / L+TDZ0.1mg / L+EDTA+ defoamer, and inoculate 70ml in each bottle Solution, after inoculation, placed in rotary agitation and aerated culture, added 30g / L sucrose, pH 5.8, light 1500lx, temperature 15°C, the cells cultured in suspension were placed on a solid plate, and cell lines with high content of secondary metabolites were screened.

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PUM

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Abstract

The invention researches a fast propagation method for arachniodes exilis callus tissues and suspension cells. The method comprises the following steps: obtaining sterile leaves; inducing the callus tissues; culturing the suspension cells and the like. The method provided by the invention can be used for quickly obtaining massive arachniodes exilis cells, is simple and controllable in operation, high in cell reproductive rate and short in growing period, and can be used for providing infinite pharmaceutical raw materials, so that the medicinal components of arachniodes exilis cells are effectively developed and utilized favorably.

Description

technical field [0001] The invention relates to the cultivation of suspension cells of Auricularia compound leaf under the condition of tissue culture, and belongs to the technical field of plants. Background technique [0002] Auricularia compound, Arachniodes exilis (Hance) Ching, Lepidopteridaceae, mainly distributed in tropical and subtropical regions, in my country, it is distributed in Jiangsu, Anhui, Zhejiang, Jiangxi, Fujian, Taiwan, Guangdong, Sichuan, Guizhou, Yunnan and other places. Most of them grow in mixed forests in valleys ranging from hundreds to thousands of meters above sea level. It likes a cool, humid environment and well-drained sandy loam, likes diffuse light, strong shade tolerance, and a suitable growth temperature of 14-20°C. Auricularia compound is also a kind of traditional Chinese medicine, which is used for clearing heat, detoxifying, and treating sores. At present, its use is mainly collected in the wild, without large-scale promotion and pl...

Claims

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Application Information

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IPC IPC(8): C12N5/04C12N5/02
Inventor 杨存
Owner NANJING TONGZE AGRI SCI & TECH
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