Method for improving content of betulin and oleanolic acid in birch cell by utilizing MeJA (methyl-jasmonate) and SA (salicyl acid)

Abstract

The invention belongs to the field of biological engineering, and particularly relates to a method for improving the content of betulin and oleanolic acid in a birch cell by utilizing MeJA (methyl-jasmonate) and SA (salicyl acid), which comprises the following steps of: inoculating a birch tissure culture seedling stem to NT + 0.1 mg / L of 6-BA and 0.01 mg / L of TDZ culture mediums to induce birch callus, and obtaining a birch suspension cell with high content of total triterpenes through multi-time subculture, screening and suspension culture; inoculating the screened cell into the same liquid culture medium, wherein the inoculation amount is 3g / 100ml; adding 100mu Mum / L of MeJA and 50mg / L of SA when the shake culture is performed at the seventh day; after 24-48h, obtaining the product; and detecting the content of total triterpenes, betulin and oleanolic acid in the cell. Through the comparison of the two kinds of processing methods, the content of total triterpenes is improved by 21.6-31.7%, the content of oleanolic acid is improved by 277.4-369.34%, and the content of betulin is improved by 44.42-179.58%. The application of the invention provides a new path for solving the problem of shortage of natural plant drug resources and realizing large-scale industrial production of betulin and oleanolic acid drugs.

Description

Technical field

[0001] The invention belongs to the field of bioengineering, and in particular relates to a method for improving betulin alcohol and oleanolic acid in birch cells by utilizing MeJA and SA. Background technique

[0002] There are important triterpenoids in birch bark. Among them, betulin and oleanolic acid have the effect of inhibiting various tumors and HIV. Because of their natural, high efficiency and low toxicity, they are regarded as excellent and have great potential for development. Pioneering drugs against AIDS and tumors. However, the tree has a long growth cycle. If betulin is obtained, the trees need to be felled or peeled, which will destroy resources. In addition, its complex structure, low synthetic efficiency by artificial methods, and high cost have become the main reasons that restrict its large-scale clinical application. . The plant cell culture technology established on the basis of cell totipotency provides a new way for the production o...

Examples

Embodiment Construction

[0009] 1) Source of plant material

[0010] The white birch mother tree was taken from the Northeast Forestry University White Birch Enhanced Seed Garden. The dormant buds and seeds of the 7-8-year-old grafted excellent tree (30-year-old scion) were used as explants to induce tissue culture seedlings (WPM+1.0mg / L 6-BA medium). ) tissue culture seedlings of birch. The stems of birch tissue culture seedlings were used as explants, NT was used as the basic medium, and 0.1 mg / L6-BA and 0.01 mg / LTDZ were added to induce callus. After passage, it was transferred to liquid medium for suspension culture. After more than 20 generations of transfer, a stable suspension culture cell system was established. The suspension culture subculture period was 15d, and the inoculation amount was 3g cells in 100mL liquid medium.

[0011] 2) Culture conditions

[0012] Solid medium: NT medium+0.1mg / L 6-BA+0.01mg / LTDZ, sucrose 20g / L, agar powder 5.3g / L, pH value 6.0-6.5. Liquid medium is solid me...

Technical field

[0001] The invention belongs to the field of bioengineering, and in particular relates to a method for improving betulin alcohol and oleanolic acid in birch cells by utilizing MeJA and SA. Background technique

[0002] There are important triterpenoids in birch bark. Among them, betulin and oleanolic acid have the effect of inhibiting various tumors and HIV. Because of their natural, high efficiency and low toxicity, they are regarded as excellent and have great potential for development. Pioneering drugs against AIDS and tumors. However, the tree has a long growth cycle. If betulin is obtained, the trees need to be felled or peeled, which will destroy resources. In addition, its complex structure, low synthetic efficiency by artificial methods, and high cost have become the main reasons that restrict its large-scale clinical application. . The plant cell culture technology established on the basis of cell totipotency provides a new way for the production o...

Examples

Embodiment Construction

[0009] 1) Source of plant material

[0010] The white birch mother tree was taken from the Northeast Forestry University White Birch Enhanced Seed Garden. The dormant buds and seeds of the 7-8-year-old grafted excellent tree (30-year-old scion) were used as explants to induce tissue culture seedlings (WPM+1.0mg / L 6-BA medium). ) tissue culture seedlings of birch. The stems of birch tissue culture seedlings were used as explants, NT was used as the basic medium, and 0.1 mg / L6-BA and 0.01 mg / LTDZ were added to induce callus. After passage, it was transferred to liquid medium for suspension culture. After more than 20 generations of transfer, a stable suspension culture cell system was established. The suspension culture subculture period was 15d, and the inoculation amount was 3g cells in 100mL liquid medium.

[0011] 2) Culture conditions

[0012] Solid medium: NT medium+0.1mg / L 6-BA+0.01mg / LTDZ, sucrose 20g / L, agar powder 5.3g / L, pH value 6.0-6.5. Liquid medium is solid me...