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Method for preparing soybean isoflavonoid with tissue culture

A soybean isoflavone and tissue culture technology, which is applied in horticultural methods, botanical equipment and methods, applications, etc., can solve problems such as high requirements for operators, differences in rooting efficiency, and difficult methods to master.

Inactive Publication Date: 2008-12-10
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The hairy roots obtained by this method need to be sterilized several times before expanding the culture, the process is cumbersome, the requirements for operators are high, and the method is not easy to master
In addition, different Agrobacterium rhizogenes have different infecting abilities to different soybean varieties, resulting in great differences in the efficiency of producing hairy roots. In addition to the difference in operators, the stability of this method of producing hairy roots is affected.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] 1. Select high-quality soybean varieties, take healthy and mature soybean seeds, sterilize them with chlorine gas for 2 to 10 hours, inoculate the soybean hilum down into MS solid medium, and the pH value of the medium is 5.5. The soybean seeds inoculated on the culture medium are cultured at 24°C for 7 days to obtain healthy soybean sterile seedlings.

[0023] 2. cut the true leaves of soybean sterile seedlings, inoculate in the callus induction medium (the inorganic component of MS solid medium, add the organic component of B5 solid medium, add concentration in this mixed solid medium simultaneously: 0.1 mg / L of 6-benzylaminoadenine and 1.0 mg / L of 2,4 dichlorophenoxyacetic acid, the balance being agar and sterilized water), subcultured once every 30 days. Cultivate for 90 days at a culture temperature of 24°C.

[0024] 3. Take out the callus, break it up, and extract it with 70% ethanol at a ratio of 5ml ethanol to 0.5g sample.

[0025] The content of daidzein and ...

Embodiment 2

[0027] 1. Select soybean varieties with high isoflavone content, select plump soybean seeds that are free from diseases and insect pests, soak soybean seeds in a supersaturated solution of bleaching powder tablets for 15 minutes, rinse them with sterile water for 3 times, and inoculate the seeds in B5 solid culture On the basis, cultured at 25°C for 10 days to obtain sterile vaccines.

[0028] 2. get the root of soybean sterile seedling, inoculate in the callus induction medium (MS solid medium, add the 6-benzylaminoadenine that concentration is 0.1mg / L and concentration be 1.0 in this solid medium simultaneously mg / L of 2,4-dichlorophenoxyacetic acid, the balance being agar and sterilized water), subcultured once every 30 days. Cultivate for 90 days at a culture temperature of 24°C to 27°C.

[0029] 3. Take out the callus, break it up, extract with 70% ethanol, and extract with the proportion of 0.5g sample and 5ml ethanol.

[0030] The content of daidzein and genistein in ...

Embodiment 3

[0032] 1. Select soybean materials with high isoflavone content, select plump soybean seeds that are free from diseases and insect pests, sterilize them with chlorine gas for 2 hours to 24 hours, inoculate the sterilized seeds into the inorganic components of MS solid medium plus B5 solid culture On the solid medium composed of basic organic components, cultured at 26°C for 9 days to obtain sterile seedlings.

[0033] 2. cut the cotyledon of soybean sterile seedling, inoculate in the callus induction medium (B5 solid medium, add the 6-benzylaminoadenine that concentration is 0.1mg / L and concentration be 1.0 in this solid medium simultaneously mg / L of 2,4-dichlorophenoxyacetic acid, the balance being agar and sterilized water), subcultured once every 30 days. Cultivate for 90 days at a culture temperature of 24°C to 27°C.

[0034] 3. Take out the callus, break it up, extract with 70% ethanol, and extract with the proportion of 0.5g sample and 5ml ethanol.

[0035] The content...

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PUM

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Abstract

The invention relates to a method for producing soy isoflavone by tissue culture in the agricultural technological field. The method comprises the following steps of: step one, the obtaining of an aseptic soy seedling; step two, the inducement of a callus tissue by an explant; and step three, the extraction of the soy isoflavone, wherein, in step one, a mature soy seedling is picked up, sterilized by a sterilizing agent, inoculated to a germination culture medium and cultured to produce the aseptic soy seedling; in step two, a tissue of the aseptic soy seedling obtained in the step one is cut off; and the tissue comprising a hypocotyledonary axis, a cotyledonary node, cotyledon, euphylla, root and other parts is inoculated to a callus tissue inducement culture medium; the tissue is subject to successive transfer culture once every thirty days for successive three times; in step three, the soy isoflavone is extracted from the callus tissue obtained in the step two by ethanol. The method has simple process, convenient and easy operation and strong stability and is not limited by season and a breeding period.

Description

technical field [0001] The invention relates to a production method in the field of agricultural technology, in particular to a method for producing soybean isoflavones by tissue culture. Background technique [0002] Soybean isoflavones (soybean isoflavones) are secondary metabolites formed during the growth of soybeans. They are one of the bioflavonoids and a kind of phytoestrogens. (daidzei II). Soy isoflavones are a very important non-nutritional ingredient in soybeans, which have anti-oxidation, anti-cancer, immune-enhancing, osteoporosis-preventing, and estrogen-like effects. The naturally occurring soybean isoflavones found so far are polyphenolic compounds, which are mainly distributed in the cotyledons and hypocotyls of soybean seeds. At present, isoflavones are extracted directly from soybeans at home and abroad; isoflavones are extracted from intermediate products and wastes after soybean processing, and soybean isoflavones are produced by decomposition and synt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D311/36A01H4/00
Inventor 曹越平陈冰周斐红朱筠
Owner SHANGHAI JIAO TONG UNIV
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