Method for production of resveratrol from polygonum cuspidatum trichoid root and enlarged cultivation

A technology of resveratrol and expanded culture, applied in the biological field, can solve problems such as unreported, staying in the experimental stage, etc., and achieve the effects of reducing production cost, saving time, and improving biomass

Active Publication Date: 2015-02-25
成都市三禾田生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, Yu Shuhong et al. have further applied for a patent for the preparation of polydatin from the hairy root of Polygonum cuspidatum, and the patent number is 200510012809.7, but it is only in the experimental stage at present, and no further research on industrialization has been carried out.
At present, the production of resveratrol from the hairy root of Polygonum cuspidatum has been reported, but there is no report on the industrialization of the production of resveratrol from the hairy root of Polygonum cuspidatum using a bioreactor

Method used

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  • Method for production of resveratrol from polygonum cuspidatum trichoid root and enlarged cultivation
  • Method for production of resveratrol from polygonum cuspidatum trichoid root and enlarged cultivation
  • Method for production of resveratrol from polygonum cuspidatum trichoid root and enlarged cultivation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] (1) Disinfection of explant 1

[0060] The young stems of Polygonum cuspidatum were selected as explants 1, and the steps were as follows: Rinse the shoots with sprouts in running water for 3 hours, soak in alcohol for 60 seconds in an ultra-clean workbench, rinse with sterile water 5 times, and soak with 0.1% mercury 5 -8min, rinse with sterile water 6 times, absorb the water with sterilized filter paper, and set aside.

[0061] (2) Induction of buds and increment

[0062] Transfer the explant 1 that has been sterilized in step (1) to the bud induction medium: MS solid basic medium + 6-BA2.5mg / L+NAA0.5mg / L+sucrose 30g / L, 7 days later The sprout point begins to grow and grows into a complete plant after 30 days. Continue subculture, subculture medium: MS solid basic medium+6-BA2.5mg / L+NAA0.5mg / L+TDZ0.05mg / L+sucrose 30g / L, to grow to a well-developed complete tissue culture of Polygonum cuspidatum After seedling, spare. Growth conditions: temperature 25±2℃, fluorescent lamp...

Embodiment 2

[0078] (1) Same as Example 1 (1)

[0079] (2) Induction of buds and increment

[0080] Transfer the sterilized stems in step (1) to the bud induction medium: MS solid basic medium + 6-BA2.0mg / L+NAA0.5mg / L+ sucrose 30g / L, and start to grow after 7 days The budding point will grow into a complete plant after 30 days. Continue subculture, subculture medium: MS solid basic medium+6-BA0.5mg / L+IAA0.5mg / L+TDZ0.05mg / L+sucrose 30g / L, wait until it grows to a well-developed complete tissue culture of Polygonum cuspidatum After seedling, spare. Growth conditions: temperature 25±2℃, fluorescent lamp, illumination 1500-2000lux, illumination time 14-16h.

[0081] (3) Induction of Polygonum cuspidatum hairy roots

[0082] Take the stems of Polygonum cuspidatum tissue cultured seedlings used in step (2) and transfer them to MS blank medium for dark culture for 2 days at a culture temperature of 25±2°C.

[0083] (4) Same as Example 1 (4)

[0084] (5) Infection and co-cultivation

[0085] The co-cultur...

Embodiment 3

[0092] (1) Same as Example 1 (1)

[0093] (2) Induction of buds and increment

[0094] Transfer the sterilized + in step (1) to the bud induction medium: MS solid basic medium+6-BA2.0mg / L+NAA0.5mg / L+sucrose 30g / L, and buds begin to appear after 7 days It will grow into a complete plant after 30 days. Continue subculture, subculture medium: MS solid basic medium+6-BA2.0mg / L+NAA0.5mg / L+sucrose 30g / L. After it grows to a well-developed intact Polygonum cuspidatum tissue culture plantlet, it is reserved. Growth conditions: temperature 25±2℃, fluorescent lamp, illumination 1500-2000lux, illumination time 14-16h.

[0095] (3)(4)(5) Same as Example 1 step (3)(4)(5)

[0096] (6) Sterilization and optimized culture

[0097] Rinse the leaves of Polygonum cuspidatum tissue cultured seedlings co-cultured in step (5) with sterile water for 3 times and transfer to sterile medium MS solid basic medium + cb300mg / L, 25℃, 14h·d -1 Sterilize culture under scattered light. Transfer once every 5 days, r...

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Abstract

The invention belongs to the field of biotechnology, relates to a method for induction of polygonum cuspidatum trichoid root and enlarged cultivation by use of a biological reactor, and in particular relates to a method for production of resveratrol from the polygonum cuspidatum trichoid root and enlarged cultivation. According to the method, polygonum cuspidatum tissue culture seedlings are induced successfully from polygonum cuspidatum plant wild stem sections with buds as explants, a polygonum cuspidatum tissue culture system is established, a polygonum cuspidatum trichoid root system is successfully induced, a polygonum cuspidatum trichoid root resveratrol production system is established, the resveratrol is successfully produced from the polygonum cuspidatum trichoid root by an airlift bioreactor, the resveratrol content is determined, and the resveratrol extracted and separated. Through the production of the resveratrol from the polygonum cuspidatum trichoid root, compared with the current mode of production, the method for the production of the resveratrol from the polygonum cuspidatum trichoid root has the rapid growth characteristic on a hormone free cultivation medium, so that the method has the advantages of simple operation, low cost, no limitation of climate, lands and other natural conditions, and the like, and a reliable source and material basis for the future industrial production is provided.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for inducing and expanding culture of Polygonum cuspidatum hairy roots by using a bioreactor, in particular to a method for producing resveratrol and expanding culture of Polygonum cuspidatum hairy roots. Background technique [0002] Resveratrol (Resveratrol, 3,4',5-trihydroxy-trans-stilbene) is trans 3,4',5-trihydroxy-stilbene. It is a stilbene compound, in free form and glycoside bound form. It exists in many plants such as Polygonum cuspidatum, grapes, peanuts, etc. Among them, Polygonum cuspidatum is the most abundant. It is an important plant antitoxin. Resveratrol has a variety of pharmacological activities, such as antibacterial, anti-oxidant, anti-cancer, anti-platelet aggregation, anti-HIV activity, etc. It can repair cellular DNA damage caused by atypical pneumonia prescriptions, especially the enemy of human health- -The treatment and prevention of tumors and heart di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/22C12N5/04A01H4/00C12R1/01
CPCA01H4/00C12N5/04C12P7/22
Inventor 洪汉君熊小灿
Owner 成都市三禾田生物技术有限公司
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