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Method capable of effectively inhibiting browning for multiplication culture of buckwheat callus

A callus, proliferation culture technology, applied in horticultural methods, botanical equipment and methods, plant regeneration and other directions, can solve the problems of browning culture materials, browning of culture materials, easy to produce oxidation, etc. The effect of reducing the browning rate

Active Publication Date: 2012-12-19
CHENGDU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Because buckwheat callus contains a large amount of flavonoids, it is very easy to produce oxidation during callus culture; therefore, in the process of buckwheat callus proliferation and culture, severe browning of culture materials often occurs, so that buckwheat callus The death of the culture material due to severe browning during the mass proliferation of the tissue
There is no report about the solution

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] (1) Acquisition of buckwheat callus: select buckwheat petioles sown in spring and grow healthy, first sterilize with 0.1% mercury chloride for 4 minutes, then wash 3 times with sterile water, and then cut into 1.5cm length, Insert into the callus induction medium MS+6-BA 0.5mg / L+2,4-D 4mg / L+IAA 0.1mg / L, at 25°C, under the conditions of 10 hours of light per day and light intensity of 2000 lx Under the conditions of culture, the pH value of the medium is 6.0, sucrose 30g / L, agar 6.8 g / L;

[0017] (2) Pre-cultivation of callus: Select buckwheat callus that has been induced for 20 days without browning and grows fast, and put it in an incubator for pre-culture for 10 days at a temperature of 8°C and without light;

[0018] (3) Proliferation culture of callus: Cut the pre-cultured callus horizontally (to reduce the wound area of ​​the callus), and then quickly insert the cut end into the proliferation medium 1 / 2 MS+6-BA 1.0 mg / L+IAA 2 mg / L + citric acid 10 mg / L+ L-cysteine...

Embodiment 2

[0020] (1) Acquisition of buckwheat callus: select buckwheat petioles sown in spring and grow healthy, first sterilize with 0.1% mercury chloride for 5 minutes, then wash 4 times with sterile water, and then cut into 1.5cm length, Into the callus induction medium MS+6-BA 1.5mg / L+2,4-D 2mg / L+IAA 0.5mg / L, at 25°C, under the conditions of 14 hours of light per day and light intensity of 2000 lx Under the conditions of culture, the pH value of the medium is 6.0, sucrose 30g / L, agar 6.8 g / L;

[0021] (2) Pre-cultivation of callus: Select buckwheat callus that has been induced for 25 days without browning and grows fast, and put it in an incubator for pre-cultivation for 8 days at a temperature of 12°C and without light;

[0022] (3) Proliferation culture of callus: After cutting the pre-cultured callus, quickly insert the cut end into the proliferation medium 1 / 2 MS+6-BA 1.5 mg / L+IAA 1 mg / L + citric acid 7 mg / L+ L-cysteine ​​140 mg / L, cultured at 8°C for 3 days without light, and ...

Embodiment 3

[0024] (1) Acquisition of buckwheat callus: choose buckwheat petioles sown in spring and growing healthily, sterilize them with 0.1% mercury chloride for 4 minutes, wash them with sterile water for 4 times, cut them into 2 cm lengths, and then Put into the callus induction medium MS+6-BA 1.0mg / L+2,4-D 5mg / L, cultivate under the conditions of 25°C, 12 hours of light every day, and light intensity of 2000 lx. pH value is 6.0, sucrose 30g / L, agar 6.8 g / L;

[0025] (2) Pre-cultivation of callus: select buckwheat callus induced for 20 days without browning, and put it in an incubator for 12 days at a temperature of 6°C without light;

[0026] (3) Proliferation culture of callus: After cutting the pre-cultured callus, quickly insert the cut end into the proliferation medium 1 / 2 MS+6-BA 0.6mg / L+IAA 3mg / L +citric acid 12 mg / L+ L-cysteine ​​60 mg / L, first cultured at 6°C without light for 5 days, and then rapidly cultured at 27°C with 7 h of light per day and light intensity of 1800 l...

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Abstract

The invention discloses a method capable of effectively inhibiting browning for the multiplication culture of buckwheat callus, which is characterized in that before the multiplication culture, the buckwheat callus is pre-cultured, and citric acid and L-cysteine are added into a multiplication culture medium. Therefore, the browning rate of the buckwheat callus during the multiplication culture is effectively inhibited, the multiplication times of the buckwheat callus is increased, and a new method is provided for greatly and quickly producing flavonoids (rutin).

Description

technical field [0001] The invention relates to a method for the proliferation and culture of buckwheat callus, in particular to a method for the proliferation and culture of buckwheat callus which effectively inhibits browning. Background technique [0002] Buckwheat (Fagopyrum esculentum) is a dicotyledonous crop belonging to the genus Fagopyrum Mill. in the family Polygonaceae, and is an important medicinal and food homologous crop. The content of rutin in buckwheat is relatively high. As the main component of flavonoids, rutin can not only be used as an antioxidant and a raw material for functional foods, but also reduce capillary fragility and improve microcirculation. It is mainly used clinically for diabetes, high Adjuvant therapy for blood pressure etc. At present, the main raw material for extracting rutin in China is Sophora japonica; however, due to the low yield of Sophora japonica and scarcity of resources, it cannot fully meet the market demand. Therefore, pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 王跃华赵钢孙雁霞宋超陈丽段有丽邬晓勇邹亮彭镰心
Owner CHENGDU UNIV
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