Method for preparing polygonin through tissue culture and inducement of hairy roots of giant knotweed

A hairy root and polydatin technology, applied in the field of plant biology, can solve the problems of slow cell growth, hormone-dependent maintenance, poor genetic stability, etc., and achieve the effects of simple extraction operation, short production cycle, stable quality and yield

Inactive Publication Date: 2006-04-19
HEBEI MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are many disadvantages in the large-scale production of secondary metabolites in cell culture, such as slow cell growth and hormone-dependent maintenance, low content, poor genetic stability, and sensitivity to shear stress during amplified culture, etc.

Method used

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  • Method for preparing polygonin through tissue culture and inducement of hairy roots of giant knotweed
  • Method for preparing polygonin through tissue culture and inducement of hairy roots of giant knotweed
  • Method for preparing polygonin through tissue culture and inducement of hairy roots of giant knotweed

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] 1. Select the young buds at the top of the stems of wild Polygonum cuspidatum plants that are healthy, well-developed, free of diseases, insect pests, and fungi. After routine disinfection, place them in a medium that induces clustered bud differentiation: MS+0.05mg / LTDZ+0.5mg / L NAA+0.1mg / L 6-BA. The culture conditions are daytime temperature: 27±1℃, night temperature: 20±1℃; humidity: 60-65%, light intensity: 28μmol / m 2 .s, light time: 14h / d. After 15 days, the clustered seedlings were separated into individual plants, and placed in the growth medium of strong seedlings: MS+0.1mg / L CPPU+1.0mg / LNAA, and the culture conditions were the same as above. One week later, transplant single tissue cultured seedlings into rooting medium: MS+1.0 mg / L NAA+0.5% activated carbon, and the culture conditions are the same as above. The adventitious bud organ proliferation pathway can be used for sustainable rapid propagation, the reproduction coefficient of each bud is 5-7, and a we...

Embodiment 2

[0050] In the step (2), cut the stem section of the aseptic seedling of Polygonum cuspidatum as the infection material. In step (3), at the same time, add 500 μl of Agrobacterium tumefaciens C58 bacterial solution to 100ml YEB culture solution, and Agrobacterium rhizogenes ATCC11325 at 25°C, 140r min -1 , Dark co-cultivation for 22h before use. In step (4), the stem segment explants were soaked in the dual bacteria activation solution for 10min, and placed in MS under dark conditions. 0 CCP cultivated. In step (5), PCR molecular identification is carried out after separating and culturing the hairy roots obtained by stem segment induction; in step (6), the hairy roots obtained by stem segment induction are optimized and cultivated to obtain a large number of proliferating hairy root strains . In step (7), the products of polydatin and resveratrol are extracted, separated and purified from the hairy roots induced by stem segments. In step (8), the contents of polydatin and ...

Embodiment 3

[0052] In the step (2), cut out the sprouts of the aseptic seedlings of Polygonum cuspidatum as infection materials. In step (3), Agrobacterium rhizogenes ATCC11325 was heated at 26°C and 150r min -1 , and used after 24 hours of dark co-cultivation. In step (4), the young shoot explants were soaked in 100ml of activated bacterial liquid, treated in an ultrasonic oscillator for 5-10 sec, and after taking out and absorbing excess bacterial liquid, placed in MS under dark conditions. 0 CCP cultivated. In step (5), PCR molecular identification is carried out after separating and culturing the hairy roots obtained by the sprout induction; in step (6), the hairy roots obtained by the sprout induction are optimized and cultivated to obtain a large amount of proliferating hairy root strains . In step (7), the products of polydatin and resveratrol are extracted, separated and purified from the hairy roots induced by the young shoots. In step (8), the contents of polydatin and resve...

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Abstract

A process for preparing the polygonoside from the trichiform root of rhizome polygoni cuspidati by tissue culture and inducing includes such steps as tissue culture of wild rhizome polygoni cuspidati, activating culture of Agrobacterium rhizogene, inducing the trichiform root of rhizome polygoni cuspidati by the T-DNA fragment on rooting plasmide of Agrobacterium rhizogene, optimalizing culture of said trichiform root, and extracting polygonoside from said trichiform root.

Description

technical field [0001] The invention relates to a method for preparing polydatin by tissue culture-induced hairy roots of Polygonum cuspidatum, belonging to the field of plant biotechnology. Background technique [0002] Polydatin (polydatin) and resveratrol (resVeratrol) are both stilbene polyphenols naturally produced in plants. They are monomeric compounds with clear chemical structures and definite curative effects. Inhibiting platelet aggregation, lowering blood lipids, anti-virus and enhancing the body's immunity, and more importantly, it can specifically kill cancer cells without affecting normal cells. It is known as "the most promising drug in the 21st century". Therefore, it is widely used in clinical medicine and health care industry, and there is a great demand in domestic and foreign markets. [0003] Polygonadin and resveratrol exist in many plants in nature, such as grapes, peanuts, hellebore, knotweed, European spruce, etc. Among them, the content in knotwee...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00C07H15/203C07H1/08
Inventor 于树宏范庆书查建蓬张嫡群
Owner HEBEI MEDICAL UNIVERSITY
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