Method for scaled production of radix astragali hairy roots
A hairy root and large-scale technology, applied in the field of astragalus biotechnology cultivation, can solve the problems of astragalus reduction, heavy metal pollution, and pesticide reduction in cultivated varieties, and achieve the effect of short production cycle and uniform production quality
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Embodiment 1
[0059] A method for large-scale production of astragalus hairy roots, comprising the following steps:
[0060] Step 1. Prepare Astragalus aseptic vaccine
[0061] Soak Astragalus mongolica seeds in tap water for 24 hours, soak in 70% ethanol for 3 minutes, rinse with sterile water for 5 times; pour 1% mercuric chloride solution for 10 minutes; rinse with sterile water for 5 times; inoculate until pre-sterilized cultured on MS medium at a temperature of 25°C and a humidity of 60%, to obtain aseptic Astragalus seedlings;
[0062] Step 2. Strain activation and cultivation
[0063] Add Agrobacterium ATCC15834 into liquid YEB medium and cultivate at 26°C at -250rpm until the OD600 value is 0.6; then, culture the bacteria in 1 / 2MS liquid medium at 26°C and 150rpm for 30min;
[0064] Step 3. Material pre-cultivation and co-cultivation
[0065] The leaves of the aseptic seedlings are transferred to the MS medium that adds 0.3mg / L of the phytohormone IBA, cultured in the dark for 2 ...
Embodiment 2
[0072] A method for large-scale production of astragalus hairy roots, comprising the following steps:
[0073] Step 1. Prepare Astragalus aseptic vaccine
[0074] Soak the Astragalus membranaceus seeds in tap water for 20 hours, soak them in 70% ethanol for 2 minutes, rinse them with sterile water for 5 times, pour them into 1% mercuric chloride solution and soak them for 8 minutes, rinse them with sterile water for 5 times, and inoculate them until they are extinguished in advance. Bacteria on the MS medium, temperature 30 ℃, humidity 60% culture, to obtain Astragalus aseptic vaccine;
[0075] Step 2. Strain activation and cultivation
[0076] Add Agrobacterium LBA9402 into liquid YEB medium and cultivate at 26°C and 220rpm until the OD600 value is 0.5; then, culture the bacteria liquid in 1 / 2MS liquid medium and cultivate at 28°C and 180rpm for 30min;
[0077] Step 3. Material pre-cultivation and co-cultivation
[0078] The petiole of the aseptic seedling was transferred ...
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