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Suspension culture method for cottonrose hibiscus cells

A technology of suspension culture and cell suspension, which is applied in the field of suspension culture of hibiscus cells, to achieve the effects of increasing cell growth speed, increasing cell fresh weight, and high induction rate

Inactive Publication Date: 2018-05-15
HANGZHOU NIUBEI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The establishment of the suspension cell line of Hibiscus hibiscus has not been reported yet. In industrial production, culturing hibiscus hibiscus cells is one of the effective ways to obtain secondary metabolic drugs and extract pharmacologically active factors of hibiscus hibiscus cells, and can study the growth, development and differentiation of hibiscus hibiscus The mechanism and the law of genetic variation are of special significance to the genetic breeding of Hibiscus hibiscus

Method used

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  • Suspension culture method for cottonrose hibiscus cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] (1) Acquisition of aseptic explants: choose hibiscus hibiscus axillary buds with healthy growth and 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 hibiscus hibiscus callus: Insert the axillary buds after disinfection into the callus induction medium, and culture them in the dark for 10 days at 23° C., then light 8 hours every day and light intensity is 1000 lx. For culturing, the callus induction medium is to add KT (i.e. 6-furfurylaminopurine) 0.1mg / L, 6-BA (i.e. 6-benzylaminoadenine) 0.1mg / L, 2,4- D (i.e. 2,4...

Embodiment 2

[0035] The influence of embodiment 2 acid hydrolyzed casein on Hibiscus hibiscus cell suspension culture

[0036] Select hibiscus single cells or loose cell aggregates in the middle and upper part of the suspension culture medium after pre-cultivation and transfer them to the cell suspension medium with different hormones added. Other operations are the same as in Example 1. The results of hibiscus cell suspension culture are shown in Table 1. Show. The results showed that the culture medium with different hormone combinations could induce hibiscus hibiscus cell proliferation to varying degrees, but when acid hydrolyzed casein was added to the culture medium, the cell browning rate decreased, the cell growth multiple increased, and the cell growth rate also increased, among which adding 0.6 g / L acid hydrolyzed casein, the cell growth rate was the fastest, reaching 3.37g / (d·L), and the cell proliferation multiple reached 2.98.

[0037] Table 1 Effect of acid hydrolyzed casein ...

Embodiment 3

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

[0040] The non-brown yellow-white callus of hibiscus hibiscus obtained in step (2) of embodiment 1 is transferred into the pre-culture liquid medium and carried out pre-cultivation of different days, and other steps are the same as in embodiment 1, and the results of suspension culture of hibiscus hibiscus cells are shown in Table 2 shows. The results showed that after pre-cultivation for 7 days, the biomass of Hibiscus hibiscus cells subcultured for 3 generations proliferated the most, the cell proliferation multiple was 3.24 times, and the cell growth rate was 3.15g / (d·L).

[0041] The impact of table 2 pre-cultivation on hibiscus cell suspension culture

[0042] Pre-cultivation time (days)

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Abstract

The invention discloses a suspension culture method for cottonrose hibiscus cells. The method comprises the following steps: preparing cottonrose hibiscus axillary buds as an explant; disinfecting thesurface; placing the disinfected explant on a callus induction medium; culturing to obtain callus; and then performing cell pre-culturing, cell suspension culture and cell subculture to obtain a large number of suspension culture cells. According to the method, the cottonrose hibiscus cells are subjected to suspension culture, so that the cottonrose hibiscus cell culture products with high effective component content can be massively and quickly produced, so that a good foundation is provided for producing secondary metabolic products through cottonrose hibiscus cell suspension culture.

Description

(1) Technical field [0001] The invention relates to the field of biotechnology, in particular to a method for suspension culture of hibiscus hibiscus cells. (2) Background technology [0002] Hibiscus (Hibiscus mutabilis Linn.) is a perennial small tree or deciduous shrub of the Malvaceae Hibiscus genus. It is cultivated in the north and south of our country. Hibiscus hibiscus is not only an important ornamental garden flower, but also an important medicinal plant. The leaves, flowers and roots of hibiscus hibiscus can be used as medicine, which has important medical value. Hibiscus cell tissue contains medicinal ingredients rich in flavonoids, hyperoside, inulin, phenols, reducing sugars, etc. Hibiscus has the functions of clearing heat and detoxification, reducing swelling and discharging pus, cooling blood and stopping bleeding, and regulating menstruation and hemostasis. Studies have shown that the alcohol extract of hibiscus hibiscus has a strong inhibitory effect on ...

Claims

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Application Information

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IPC IPC(8): C12N5/04C12N5/02
CPCC12N5/0025C12N5/04
Inventor 刘军谢丹
Owner HANGZHOU NIUBEI BIOTECH
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