Alfalfa endophytic pediococcus pentosaceus EL5 and application thereof
A technology of Pediococcus pentosaceus and alfalfa, applied in the application, bacteria, microorganism and other directions, can solve the problems of poor adaptability, low activity, restricting alfalfa silage, etc., and achieves the effects of low cost, easy utilization, and improved fermentation quality
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[0036] An embodiment of the present invention provides a method for preparing silage, comprising: mixing the silage raw material with the above-mentioned Pediococcus pentosaceus EL5 and fermenting to obtain the silage.
[0037] In one embodiment, the silage material can be alfalfa, and further can be alfalfa.
[0038] In the silage of one embodiment, the ratio of the viable count of Pediococcus pentosaceus EL5 to the mass of the silage material is 1.0×10 5 CFU / g~2.0×10 6 CFU / g, e.g. 2.0×10 5 CFU / g, 5.0×10 5 CFU / g, 8.0×10 5 CFU / g, 1.0×10 6 CFU / g, 1.5×10 6 CFU / g.
[0039] A method for preparing silage according to an embodiment of the present invention includes the following steps:
[0040] S1: Chop the alfalfa raw materials and mix them evenly;
[0041] S2: Add Pediococcus pentosaceus EL5 to the alfalfa raw material, and the effective viable count of Pediococcus pentosaceus EL5 per gram of alfalfa raw material is 1.0×10 6 more than one.
[0042] S3: The alfalfa system...
Embodiment
[0054] Isolation and culture of endophytic Pediococcus pentosaceus EL5
[0055] Put the fresh alfalfa material in a sterile sampling bag and bring it back to the laboratory. Weigh 10g of the whole alfalfa plant in the sterile environment of the ultra-clean workbench and then carry out surface disinfection operation: immerse it in 70% ethanol for 90s, 3.25 ~4% sodium hypochlorite solution for 120s, 70% ethanol for 30s, then rinsed with sterile water for 3 times and used sterile filter paper to absorb the surface moisture, and the sterile water of the last rinse was applied as a control.
[0056] Put the sterilized material into a sterilized mortar for grinding (add a little sterilized quartz sand to help grinding, and calcium carbonate as a buffer), grind the alfalfa tissue, add 9 mL of 8.5% sterile physiological saline and stir well, Then let stand for 3min, take the supernatant (recorded as 10 -1 Concentration gradient) 1mL and successively diluted to 10 by 10-fold dilution ...
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