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Method for producing verbascoside by using hairy roots of rehmannia

A technology of verbascoside and hairy root, applied in the field of production of verbascoside, which can solve the problems of large variation range, content difference, and uneven quality of rehmannia glutinosa, and achieve the effects of short cultivation period, easy operation, and increased yield

Active Publication Date: 2016-11-23
HENAN AGRICULTURAL UNIVERSITY
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Problems solved by technology

However, the actual situation is that the content of acteoside between different varieties of rehmannia glutinosa is significantly different. Even if the same variety is planted in different regions, there are also differences in content, and the range of variation is large, which leads to uneven quality of rehmannia glutinosa. Application restrictions and other issues
Using genetic engineering technology to obtain rehmannia glutinosa varieties with high verbascoside content is a potential and effective solution, but so far, the molecular genetic mechanism of rehmannia actcoside biosynthesis is still unclear

Method used

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  • Method for producing verbascoside by using hairy roots of rehmannia
  • Method for producing verbascoside by using hairy roots of rehmannia
  • Method for producing verbascoside by using hairy roots of rehmannia

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Embodiment 1

[0026] A method for producing verbascoside by utilizing rehmannia root hairy roots, comprising the following steps:

[0027] 1) Induction and identification of the hairy root of Rehmannia glutinosa

[0028] a. Induction of rehmannia hairy roots

[0029] Cut the leaves of Huai Dihuang sterile seedlings into about 1cm in the ultra-clean workbench 2 Place the small piece of activated Agrobacterium rhizogenes ACCC 10060 in the dipping medium (MS liquid medium + 100 μmol / L acetosyringone), take it out after 10 minutes, and blot the bacterial solution on sterile filter paper; Leaves were placed upside down in co-cultivation solid medium (MS solid medium + 100 μmol / L acetosyringone), and co-cultured at 26 °C in the dark for 2 days; after being washed with sterile water, the leaves were inoculated on selection medium (MS Solid medium + 500 mg / L cephalosporin + 100 μmol / L acetosyringone), culture in dark at 26°C, subculture once every 10 days, until hairy roots grow (see figure 1 a,...

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Abstract

The invention discloses a method for producing verbascoside by using hairy roots of rehmannia, and belongs to the technical field of bio-culture. The method comprises the following step: putting the hairy roots of the rehmannia into a salicylic acid-containing culture solution for inducing culture to obtain the verbascoside. The theory of the method is that the hairy roots, which are treated by salicylic acid, of the rehmannia can enhance the expression of key biosynthetic enzymes, such as phenylalnine ammonialyase (PAL), 4-coumaroyl-co-A ligase (4CL), cinnamate 4-hydroxylase (C4H), coumaric acid hydroxylase (C3H), copper amine oxidase (CuAO) and tyrosine carboxy-lyase (TyDC), for the verbascoside within a short time period (12 to 24 h) so as to complete gathering of the verbascoside and further increase the yield of the verbascoside. The method is easy and convenient to operate, short in culture period, convenient for industrial, large-scale and standardized implementation, and has a great significance for promotion of large-scale production of the verbascoside.

Description

technical field [0001] The invention relates to a method for producing verbascoside by utilizing the hairy roots of rehmannia glutinosa, and belongs to the technical field of biological cultivation. Background technique [0002] Rehmannia glutinosa L. is a perennial herb of the family Scrophulariaceae. Its roots are used as medicine. It is one of the famous "four medicines" in my country. Bulk medicinal materials are one of the major varieties recommended for research and utilization in the composition of the modern Chinese medicine industry chain in my country. [0003] Actascoside is one of the index components of Rehmannia glutinosa, which has biological activities such as anti-oxidation, immune regulation, anti-inflammation, liver protection, memory enhancement, and anti-tumor, with less toxic and side effects. The content of acteoside in rehmannia root is relatively low, and "Chinese Pharmacopoeia" stipulates that the content of acteoside in raw rehmannia root should no...

Claims

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Application Information

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IPC IPC(8): C12P19/44C12R1/01
CPCC12P19/44
Inventor 王丰青索艳飞张重义魏荷孙红正智惊宇谢彩侠李烜桢李娟黄勇杜家方
Owner HENAN AGRICULTURAL UNIVERSITY
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