A kind of Kouterella huagui and microbial bacterial agent and their application
A technology of Kurtella huagui and microbial inoculum, which is applied in the field of agricultural biology and can solve the problems of poor malachite green effect and the like
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[0018] Wherein, the preparation method of the microbial agent may include: respectively inoculating Kouterella huagui, Bacillus subtilis, Bacillus licheniformis and Bacillus amyloliquefaciens in culture medium for cultivation. Wherein, Kourtella huaguii, Bacillus subtilis, Bacillus licheniformis and Bacillus amyloliquefaciens can be cultured separately in independent culture systems, and the microorganisms cultured separately can be mixed in proportion. Preferably, by cultivating in separate culture systems and mixing after culturing, the total number of live bacteria per gram of microbial agent obtained is 2-10×10 7 CFU.
[0019] According to the present invention, the cultivation method of Bacillus subtilis is not particularly limited and known to those skilled in the art. For example, Bacillus subtilis can be cultivated in seed medium (LB medium) to a bacterial density of OD 600 The value is 0.6-0.8, to obtain the bacterial liquid, and then inoculate 2-5 parts by weight of...
Embodiment 1
[0030] This example is used to illustrate the present invention K. huagui and its properties.
[0031] Inoculate Kurthiahuakuii (Kurthiahuakuii) with the preservation number CCTCCNO.M2013504 into LB medium (containing tryptone 10g / L, yeast extract 5g / L, NaCl 10g / L), and culture it until the bacterial density is OD 600 The value is 0.6-0.8, and the bacterial liquid is obtained.
[0032] Malachite green (purchased from SIGMA Company) was added to the LB medium in such an amount that the concentration of malachite green was 100 mg / L to obtain the malachite green removal test medium.
[0033] Add 3mL of the above bacterial solution into 150mL of malachite green removal test medium, culture at 30°C with shaking at 180 rpm, take samples every 4h according to "GB / T20361-2006 Malachite green and crystal violet residues in aquatic products Quantitative Determination of High Performance Liquid Chromatography Fluorescence Detection Method" Determination of the concentration of malachite...
Embodiment 2
[0043] This embodiment is used to illustrate the microbial inoculant of the present invention and its performance.
[0044] (1) Inoculate 5 parts by weight of Bacillus subtilis bacteria liquid (purchased from China Agricultural Microorganism Culture Collection Center) in 100 parts by weight of culture medium (glucose 5g, soybean meal 30g, peptone 2g, distilled water 1000ml, pH7.0-7.2) And the number is ACCC03221, cultivated in the same LB medium as above to the bacterial density OD 600 The value is 0.6-0.8, to obtain the bacterial liquid), cultivated at 37°C, and samples were taken during the cultivation process and observed by direct counting under a microscope until the number of viable bacteria of Bacillus subtilis was 2×10 7 CFU / gram of medium.
[0045] (2) Inoculate 5 parts by weight of Bacillus licheniformis solution (purchased From the China Agricultural Microbiology Culture Collection Center and the number is ACCC02698, cultured in the same LB medium as above to a ba...
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