Atractylodes macrocephala special microbial agent and preparation method thereof
The technology of microbial inoculum and Atractylodes macrocephala is applied in the field of special microbial inoculum of Atractylodes Rhizoma and its preparation, which can solve the problems of carcinogenic risk and other problems, and achieve the effects of reducing pesticide residues, improving the quality of Atractylodes macrocephala, and reducing budding and flowering.
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[0035] The invention relates to a method for preparing a special microbial agent for Atractylodes macrocephala, which is obtained by mixing and fermenting the bacterial strains contained in the microbial agent or separately fermenting each bacterial strain and then mixing the fermented liquid.
[0036] In the above scheme, the method of mixed fermentation is adopted to simplify the production process and further improve the growth benefit.
[0037] In a preferred solution, the following steps are also included:
[0038] Step 1, the strain is activated to obtain the activated strain.
[0039] Step 2, primary seed culture, inoculate the activated strain into the primary seed medium, shake culture at 28-35°C, 180-200r / min for 16-20h to obtain the primary seed, wherein the primary seed medium includes the following Components in parts by weight: tryptone 4.5-5.5, yeast extract 2-3, glucose 0.5-1.5 and water 900-1100, pH 7.0-7.2.
[0040] Step 3, secondary seed cultivation, inocu...
Embodiment 1
[0047] 1) Inoculate Bacillus subtilis ACCC19742, Bacillus amyloliquefaciens CGMCC1.15674 and methylotrophic Bacillus ACCC19730 into slant medium for activation respectively, the slant medium is NA, LB or PCA medium, culture at 28-35°C for 24-36h , Obtain slanted seeds, that is, activated strains.
[0048] 2) The activated strains of the three strains were respectively subjected to primary seed culture to obtain primary seeds.
[0049] 3) Further inoculate the primary seeds into 500mL Erlenmeyer flasks containing 100-150mL secondary seed culture medium according to the inoculation amount of 5% by volume, and vibrate at 30°C and 200r / min for 10h to obtain the secondary seeds. The secondary seed medium formula is: 5g glucose, 8g soluble starch, 20g soybean meal, 1.0g calcium carbonate, 1000mL water, pH 7.0-7.2.
[0050] 4) The secondary seeds of each strain were inoculated into 15L fermenters equipped with 9L fermentation medium according to the volume percentage of 8% inoculum,...
Embodiment 2
[0053] 1) Bacillus subtilis ACCC19742, Bacillus amyloliquefaciens CGMCC1.15674 and methylotrophic Bacillus ACCC19730 were respectively inoculated for activation on slant medium to obtain activated strains. The culture medium and culture conditions refer to Example 1.
[0054] 2) The activated strains of the 3 strains were respectively subjected to primary seed culture to obtain primary seeds, and further inoculated primary seeds into secondary seed medium to obtain secondary seeds. Wherein the liquid expansion cultivation method is with reference to embodiment 1.
[0055] 3) Mix the secondary seeds of each bacterial strain in an equal volume ratio to obtain mixed seeds, and inoculate the mixed seeds in a 15L fermenter equipped with 9L fermentation medium according to a volume percentage of 10% inoculum, with a ventilation rate of 0.5vvm, 350rpm stirring, and 30°C After 24 hours of fermentation, the temperature was raised to 37° C. and the fermentation was continued for 12 hou...
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