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Method for increasing vinblastine content in vinca rosea cells

The technology of vinblastine and vinblastine is applied in the field of increasing the content of vinblastine in vinca cells, can solve the problems of long cultivation period and high cultivation cost of plant cells, and achieve the effects of shortening the induction period, avoiding browning and increasing the biosynthesis amount.

Inactive Publication Date: 2019-03-12
覃家日
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although plant cell culture can effectively improve the output of vinblastine, the plant cell culture period is longer and the culture cost is higher

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] A method for improving vinblastine content in vinca cells, comprising the following steps:

[0022] 1) Select plump periwinkle seeds, soak them in weak acid electrolyzed water for 10 seconds, wash them with sterile deionized water, and use them on the B5 foundation containing 10mg / L vitamin C, 7g / L agar and 0.1g / L oligosaccharin The aseptic seedlings were cultured on the culture medium, and the hypocotyls of the aseptic seedlings were used as explants, which were inserted into the callus induction medium for induction culture. The culture temperature was 32°C, and the light and dark cultures were alternately carried out every day. Induce the callus, then transfer the callus to the subculture medium, subculture once every 25 days, and subculture 3 times to obtain stable periwinkle callus;

[0023] 2) Select the periwinkle callus with vigorous growth, loose texture, and stable and uniform state, inoculate it into a liquid suspension medium, then add glass beads to the liq...

Embodiment 2

[0027] A method for improving vinblastine content in vinca cells, comprising the following steps:

[0028] 1) Select plump periwinkle seeds, soak them in weak acid electrolyzed water for 10 seconds, wash them with sterile deionized water, and use them on the B5 foundation containing 10mg / L vitamin C, 7g / L agar and 0.1g / L oligosaccharin The aseptic seedlings were cultured on the culture medium, and the hypocotyls of the aseptic seedlings were used as explants, which were inserted into the callus induction medium for induction culture. The culture temperature was 32°C, and the light and dark cultures were alternately carried out every day. Induce the callus, then transfer the callus to the subculture medium, subculture once every 25 days, and subculture 3 times to obtain stable periwinkle callus; wherein, the callus induction Medium is the B5 basal medium containing 1mg / L cytokinin, 0.5mg / L 6-benzylaminopurine, 30g / L sucrose, 3mg / L licorice flavone and 8g / L agar;

[0029] 2) Se...

Embodiment 3

[0033] A method for improving vinblastine content in vinca cells, comprising the following steps:

[0034] 1) Select plump periwinkle seeds, soak them in weak acid electrolyzed water for 10 seconds, wash them with sterile deionized water, and use them on the B5 foundation containing 10mg / L vitamin C, 7g / L agar and 0.1g / L oligosaccharin The aseptic seedlings were cultured on the culture medium, and the hypocotyls of the aseptic seedlings were used as explants, which were inserted into the callus induction medium for induction culture. The culture temperature was 32°C, and the light and dark cultures were alternately carried out every day. Induce the callus, then transfer the callus to the subculture medium, subculture once every 25 days, and subculture 3 times to obtain stable periwinkle callus; wherein, the callus induction Medium is the B5 basal medium containing 1mg / L cytokinin, 0.5mg / L 6-benzylaminopurine, 30g / L sucrose, 3mg / L licorice flavonoids and 8g / L agar; Add phenyla...

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PUM

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Abstract

The invention provides a method for increasing the vinblastine content in vinca rosea cells. The method comprises the following steps: (1) cultivating a vinca rosea callus; (2) cultivating a stably inherited vinca rosea cell line; and (3) inoculating the vinca rosea cell line obtained in the step (2) into a liquid suspension culture medium for induced culture to stimulate the accumulation of alkaloids in vinca rosea, wherein the induction conditions are as follows: on the 10th to 15th day of culture of the vinca rosea cell line in the liquid suspension culture medium, feeding carbon monoxide gas, adding nicotinic acid, ecdysone and silver thiosulfate into the liquid suspension culture medium, performing illumination culture with a blue LED lamp and a red LED lamp alternately, maintaining the light intensity of the blue LED lamp at 500 to 800 lx and the illumination time at 7 to 11 h / day, and maintaining the light intensity of the red LED lamp at 1,000 to 1,500 lx and the illumination time at 11 to 17 h / day till the end of the culture, wherein the vinca rosea cells with high vinblastine content is obtained. The method has the characteristics of short culture period, high vinblastineaccumulation content and the like.

Description

technical field [0001] The invention belongs to the technical field of cell culture. More specifically, the present invention relates to a method for increasing the content of vinblastine in vinca cells. Background technique [0002] Periwinkle, also known as Calendula, Sishichun, Ririxin, Yantouhong, Sanwanhua, is a plant of the genus Periwinkle in the family Apocynaceae. At present, more than 100 kinds of terpene indole alkaloids have been isolated from Vinca, such as Vinblastine, Vincristine, Serpentine, Wendoline, etc. Many alkaloids isolated from Vinca have Antitumor activity, among them, vinblastine is the alkaloid with the greatest medicinal value and the most widely used in Changchun flower. Although plant cell culture can effectively increase the output of vinblastine, the plant cell culture period is longer and the culture cost is higher. Contents of the invention [0003] As a result of various extensive and meticulous studies and experiments, the inventors o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 覃家日
Owner 覃家日