Method for improving cell proliferation efficiency and anthocyanin yield of vitis davidii

A technology for cell proliferation and thorn grapes, applied in the field of plant tissue culture, can solve the problems of slow growth, instability, low yield of secondary metabolites, etc., and achieve the effects of large light-receiving area, vigorous growth and good stability

Pending Publication Date: 2021-10-08
INST OF AGRI ENG TECH FUJIAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still many problems in plant cell culture such as slow growth, low proliferation efficiency, low yield of secondary metabolites, and instability.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] 1. Culture material: The culture material is the purple-red thorny grape cell line that has been subcultured in solid medium for a long time and has the ability to synthesize anthocyanins.

[0028] 2. For grapevine cells cultured in solid medium, select grapevine cells with uniform purple color, consistent growth state and vigorous growth.

[0029] 3. Logarithmic cell synchronous proliferation culture: the original conventional subculture time of grape thorn callus was shortened to 18 days for 20 days, that is, the callus of thorn grape callus was inoculated and cultivated to 18 days (when cultivating in solid medium for 18 days) The cell growth of the thorn grape callus is in the initial stage of logarithmic growth, and the synchronization of cell growth is high.) In the ultra-clean workbench, take an appropriate amount of callus and inoculate it into a fresh solid medium, which can make the thorn grape callus Wounded tissue cells enter a high degree of growth synchron...

Embodiment 2

[0039] 1. Culture material: The purple-red thorny grape suspension cell line that has been subcultured in liquid medium for a long time and has the ability to synthesize anthocyanins is used as the culture material.

[0040] 2. Cultivate the suspension cells of grape thorns to the middle of the exponential growth phase (cultivation to 10 days), shake the flask vigorously in the ultra-clean workbench to disperse the cell suspension evenly, and use a volume ratio of 1:8 (cell suspension Volume: the ratio of fresh liquid culture medium), pipette 5mL grapevine thorn cell suspension with a sterilized pipette, transfer to a 150mL Erlenmeyer flask containing 40mL liquid culture medium, at 25°C, light intensity 2000~ 3000Lux, photoperiod 12h light / 12h dark, shaker speed 120rpm-130rpm, shaking culture.

[0041] 3. Cultivate the suspension grapevine cells for 10 days until the mid-exponential growth phase, in the ultra-clean workbench, according to the volume area ratio of 1:30-35 (cell...

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PUM

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Abstract

The invention provides a method for improving vitis davidii cell proliferation efficiency and anthocyanin yield. The method is realized through the steps of cell line selection, vitis davidii cell suspension culture, cell solid recurrent culture, cell liquid recurrent suspension culture and the like. The vitis davidii callus which is subjected to logarithmic cell synchronous proliferation culture in a solid culture medium is inoculated into a liquid culture medium and is subjected to shaking culture in a shaking table to obtain suspension cells, or the established vitis davidii suspension cells are directly utilized to further culture the vitis davidii suspension cells to obtain vitis davidii suspension cells. A vitis davidii suspension culture cell line with highly dispersed cells, consistent growth state and uniform purple red is established, and recurrent solid culture is performed on the basis of the vitis davidii suspension culture cell line. According to the method, the advantages of solid culture and liquid culture are combined, the genetic stability and vigorous growth capacity of vitis davidii cells can be kept, a large number of vitis davidii cells rich in anthocyanin can be obtained within a short time, and the yield of anthocyanin is high; the anthocyanin can be rapidly produced on a large scale.

Description

【Technical field】 [0001] The invention relates to the technical field of plant tissue culture, in particular to a method for improving the proliferation efficiency and anthocyanin output of grapevine cells. 【Background technique】 [0002] Anthocyanin is a natural food coloring, which is safe and non-toxic. Although anthocyanins widely exist in natural plants, there are limitations in relying on natural plants to extract anthocyanins. At present, anthocyanins are mainly extracted from grape skins in the wine industry. Proanthocyanidins can be used as natural food-derived preservatives. The main sources are pine bark and grape seeds. There is an increasing demand for anthocyanins and proanthocyanidins in industries such as food processing and health products, and the original sources are not enough to meet market demand. Therefore, under the condition of "low-carbon economy", a wider range of materials must be sought to produce grape bioactive substances. Using plant cell ...

Claims

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Application Information

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IPC IPC(8): C12N5/04
CPCC12N5/04
Inventor 赖呈纯潘红赖恭梯张静黄贤贵高慧颖王琦
Owner INST OF AGRI ENG TECH FUJIAN ACAD OF AGRI SCI
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