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Buckwheat cell culture method for improving buckwheat cell synchronization

A cell culture and synchronization technology, applied in the direction of plant cells, can solve the problems of protoplasts that are not easy to fuse, and achieve the effect of promoting synthesis

Active Publication Date: 2014-04-02
CHENGDU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, if the two parental cells undergoing fusion are in different cell division stages, the protoplasts are often difficult to produce fusions

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] (1) Induction culture of buckwheat callus: Select the buckwheat seeds harvested in the same year, remove the seed coat and put them on the ultra-clean workbench, sterilize them with 0.1% mercury chloride for 3 minutes, and then rinse them with sterile water for 5 times. Inoculate in the callus culture medium of MS+2,4-D1.0mg / L+KT0.5mg / L+agar 5.5g / L+sucrose 25g / L, at a temperature of 22°C, light for 6 hours a day, and a light intensity of 800lx Under the condition of culturing, buckwheat callus was obtained;

[0014] (2) Pretreatment of buckwheat callus cells: select buckwheat callus produced in step (1) for 15 days and grow well, and insert the modified Nitsch+ 2,4-D 1.0mg / L + hydrolyzed complexin 50mg / L + glutamine 100mg / L liquid medium for suspension culture, subculture once every 25 days, and subculture twice continuously. The conditions of suspension culture are: The shaker speed is 120r / min, the temperature is 20°C, the light is 6 hours a day and the light intensi...

Embodiment 2

[0018] (1) Induction culture of buckwheat callus: select buckwheat seeds harvested in the same year, remove the seed coat and put them on the ultra-clean workbench, sterilize them with 0.1% mercury chloride for 4 minutes, and then rinse them with sterile water for 5 times. Inoculate in the callus culture medium of MS+2,4-D3.0mg / L+KT1.0mg / L+agar 6.0g / L+sucrose 30g / L, at a temperature of 25°C, light for 8 hours a day, and a light intensity of 1000lx Under the condition of culturing, buckwheat callus was obtained;

[0019] (2) Pretreatment of buckwheat callus cells: select the 20-day-old and well-grown buckwheat callus produced in step (1), and insert the modified Nitsch+ 2,4-D 2.0mg / L+hydrolyzed complexin 100mg / L+glutamine 300mg / L liquid medium for suspension culture, subculture once every 25 days, continuous subculture for 3 times, the conditions of suspension culture are: The shaking table rotates at 130r / min, the temperature is 25°C, the light is 8 hours a day and the light ...

Embodiment 3

[0023] (1) Induction culture of buckwheat callus: Select buckwheat seeds harvested in the same year, remove the seed coat and put them on the ultra-clean workbench, first sterilize with 0.1% mercury chloride for 5 minutes, and then rinse with sterile water for 5 times, Inoculate in MS+2,4-D 4.0mg / L+KT2.0mg / L+agar 7.5g / L+sucrose 35g / L callus culture medium, at a temperature of 28°C, 10 hours of light per day, and a light intensity of 1200lx Under the condition of culturing, buckwheat callus was obtained;

[0024] (2) Pretreatment of buckwheat callus cells: Take the 25-day-old and well-grown buckwheat callus produced in step (1), and insert the modified Nitsch+ 2,4-D 3.0mg / L+hydrolyzed complexin 150mg / L+glutamine 500mg / L liquid medium for suspension culture, subculture once every 25 days, continuous subculture 4 times, the conditions of suspension culture are: The shaking table rotates at 140r / min, the temperature is 28°C, the light is 10 hours a day and the light intensity is ...

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PUM

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Abstract

The invention discloses a buckwheat cell culture method for improving buckwheat cell synchronization. The method comprises the following steps: preprocessing buckwheat callus cells, inoculating the preprocessed buckwheat callus cells into different culture mediums, and performing multi-link synchronization induction treatment under different culture conditions. Therefore, the cells in different division periods can be adjusted to be in specific cell division period, the use efficiency of an inductor can be effectively improved, and the synthesis of secondary metabolites in the cultured cells is facilitated.

Description

technical field [0001] The invention relates to a buckwheat cell culture method, in particular to a buckwheat cell culture method for improving buckwheat cell synchronization. Background technique [0002] Buckwheat (Fagopyrum esculentum) is a dicotyledonous crop of Polygonaceae (Polygonaceae) buckwheat genus (Fagopyrum Mill.). Tartary buckwheat contains a large amount of flavonoids in various organs such as seeds, roots, stems, leaves and flowers. Bioflavonoids in buckwheat can regulate the permeability and fragility of capillaries, regulate blood lipids, regulate blood pressure, and resist thrombosis. etc. effect. [0003] At present, the use of plant cell protoplast fusion technology is one of the important ways to obtain plant homologous and allopolyploidy. It can effectively realize the introduction of target genes into cultivated plants, so that new varieties of cultivated plants with various excellent characters can be obtained. However, if the two parental cells u...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04
Inventor 王跃华赵钢宋超孙雁霞刘银花唐川龚婷谢珏黄菲江明殊李睿玉许志强徐恩琴梅英唐凤如
Owner CHENGDU UNIV
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