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Suspension culture method of albizia cells

A technology of suspension culture and cell suspension, applied in the directions of plant cells, tissue culture, biochemical equipment and methods, etc., can solve the problems of unprotected genetic resources of excellent traits, long cycle of sowing and breeding, and unable to meet the market demand of Albizia Julibrissin , to increase the fresh weight of cells, improve the growth rate of cells, and solve the shortage of medicinal materials.

Inactive Publication Date: 2018-05-08
HANGZHOU NIUBEI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, Albizia julibrissin is mainly propagated by sowing, and the seedling breeding cycle is very long. During the cutting propagation process, Albizia juliensis is easily infected by viruses and spreads viruses in cutting propagation, so that the genetic resources of good traits cannot be protected. The most important Affected by seasonal changes and the gradual reduction of arable land, the market demand for Albizia Juliensis cannot be met. Therefore, it is urgent to find a way and method to safely, rapidly and mass-produce cell cultures of Albizia Juliensis, and the use of tissue culture technology can not only solve the problem of Albizia Juliensis. Breeding and variation of varieties can also be mass-produced through cell culture secondary metabolites of Albizia juliensis
The establishment of the suspension cell line of Albizia juliensis has not been reported yet. In industrial production, the cultivation of Albizia juliensis cells is one of the effective ways to obtain secondary metabolite drugs and extract pharmacologically active factors of Albizia juliensis cells, and it can also study the growth, development and differentiation of Albizia juliensis. The mechanism and the law of genetic variation are of special significance to the genetic breeding of Albizia juliensis

Method used

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  • Suspension culture method of albizia cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] (1) Acquisition of aseptic explants: choose the axillary buds of Albizia julienne that grow healthy and have no damage by diseases and insect pests as explants, rinse with tap water for 30 minutes, then shake for 2 minutes in an aqueous solution with a volume concentration of 1% detergent, and use sterile Rinse with water; soak for 30s with a volume concentration of 70-75% ethanol solution on the ultra-clean workbench, and then use a mass concentration of 0.1% HgCl 2 Soak in disinfectant for 8 minutes, then rinse with sterile water for 3 times, and blot dry on sterile filter paper;

[0029] (2) Induction culture of Albizia juliensis callus: insert the sterilized axillary buds into the callus induction medium, first culture in dark at 23°C for 10 days, then light for 8 hours every day, and light intensity under the condition of 1000lx For culturing, the callus induction medium is to add 6-BA (ie 6-benzylaminoadenine) 0.1mg / L, 2,4-D (ie 2,4-dichlorophenoxyacetic acid) 0.5...

Embodiment 2

[0035] The influence of embodiment 2PVP on Albizia juliensis cell suspension culture

[0036] After pre-cultivation, single cells or loose cell aggregates of Albizia juliensis in the middle and upper part of the suspension culture medium were selected and transferred to the cell suspension medium added with different hormones. Other operations were the same as in Example 1. The results of the suspension culture of Albizia juliensis cells are shown in Table 1. Show. The results show that the culture medium of different hormone combinations can induce Albizia julienne cell proliferation to varying degrees, but after adding PVP (i.e. polyvinylpyrrolidone) in the culture medium, the browning rate of the cells decreases, the cell growth multiple increases, and the cell growth rate also increases. Among them, PVP added with 50mg / L had the fastest cell growth rate, the cell proliferation multiple reached 2.97 times, and the cell growth rate reached 3.36g / (d·L).

[0037] The effect o...

Embodiment 3

[0039] The influence of the pre-cultivation of embodiment 3 cells on the suspension culture of Albizia juliensis cells

[0040] The non-browning yellow-white callus of Albizia juliensis obtained in step (2) of Example 1 is transferred into the pre-culture liquid medium for pre-cultivation of different days, and other steps are the same as in Example 1. For the results of the suspension culture of Albizia juliensis cells, see Table 2 shows. The results showed that after pre-cultivation for 7 days, the biomass of Albizia juliensis cells subcultured for 3 generations proliferated the most, the cell proliferation multiple was 2.86 times, and the cell growth rate was 3.25g / (d·L).

[0041] Table 2 The effect of pre-cultivation on the suspension culture of Albizia juliensis cells

[0042] Pre-cultivation time (days)

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Abstract

The invention discloses a suspension culture method of albizia cells. Albizia axillary buds are extracted to be used as explants; surface sterilization is performed; then, the sterilized explants areput onto a callus induced culture medium to be cultured so as to obtain callus tissues; then, cell pre-culture, cell suspension culture and cell subculture are performed to obtain a great number of suspension culture cells. Through the cell suspension culture on the albizzia, the large-scale fast production of the albizzia cell culture materials with high active ingredient content can be realized;good foundation is laid for the secondary metabolite production through the albizzia cell suspension culture.

Description

(1) Technical field [0001] The invention relates to the field of biotechnology, in particular to a suspension culture method of albizia juliensis cells. (2) Background technology [0002] Albizia julibrissin Durazz. is a perennial deciduous tree belonging to the genus Albizia julibrissin. It is cultivated in the north and south of my country, and is distributed in East China, South China, Southwest China and Liaoning, Hebei, Henan, Shaanxi and other provinces. Albizia julibrissin is not only an important garden ornamental plant, but also an important medicinal plant. The leaves, flowers and skin of Albizia juliensis can be used as medicine, which has important medical value. Its cell tissue contains lignan glycosides, albizia juniper saponins, etc Multiple ingredients. Studies have shown that the secondary metabolites of Albizia Julibrissin tissue cells, Albizia Julibrissin Saponin, etc. have anti-fertility and anti-allergic effects, and the polysaccharide contained in Alb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04C12N5/02
CPCC12N5/04
Inventor 刘军谢丹
Owner HANGZHOU NIUBEI BIOTECH
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