Construction method of bacillus phosphorus relieving engineering bacterial strain
The technology of bacillus and engineering strains is applied in the field of construction of bacillus phosphate solubilizing engineering strains, which can solve the problems of no involvement of agricultural and environmental microorganisms, and achieve the effects of large mutation range, improved decomposition ability, and enhanced decomposition.
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Embodiment 1
[0058] App N + Ion beam implantation induces the mutation of Bacillus megaterium, and obtains an engineering strain that efficiently decomposes lecithin. Activate Bacillus megaterium on the slant of beef extract peptone medium, culture at 28-30°C for 36-48 hours, elute with normal saline, collect spores by centrifugation, dilute and spread on a sterile empty plate, and put it on the ultra-clean bench Ventilate and dry for 0.5~1h; the energy used is 15~25KeV, the dose is 40~150×10 14 ions / cm 2 N + In vacuum (vacuum degree is 2×10 -2 ~4.2×10 -3 Pa), intermittent pulse injection under sterile conditions; then the spores were eluted with physiological saline, screened on the NBRIY (containing lecithin) medium, and after identification, an engineering strain that efficiently decomposed lecithin was obtained. The ability to decompose lecithin is increased by 18-25%.
Embodiment 2
[0060] App H +Ion beam implantation induces the mutation of silicate bacteria KNP414 strain, and obtains an engineering strain that efficiently decomposes inorganic phosphorus, inorganic potassium and lecithin.
[0061] Activate KNP414 on nitrogen-free medium, culture at 28-30°C for 30-36 hours, inoculate into liquid medium without capsulated spores, culture at 28-30°C, 200-250 rpm for 48-56 hours, and collect spores by centrifugation , diluted and spread on a sterile empty plate, and dried naturally for 0.5-1h; the energy used was 15-25KeV, and the dosage was 60-200×10 14 ions / cm 2 H + In vacuum (vacuum degree is 3×10 -2 ~5×10 -3 pa), intermittent pulse injection under sterile conditions; then the spores were eluted with physiological saline, screened on NBRIY medium (containing lecithin), and then re-screened on NBRIP medium. After identification, 2 strains of efficient decomposition were obtained. Inorganic potassium, inorganic phosphorus and organic phosphorus enginee...
Embodiment 3
[0063] Apply He + Ion beam implantation induces the mutation of Bacillus subtilis to obtain an engineered strain that efficiently decomposes phytate phosphorus. Activate Bacillus subtilis on the slant of beef extract peptone medium, culture at 28-30°C for 36-48 hours, elute with phosphate buffer, collect spores by centrifugation, dilute and spread on a sterile glass slide, ultra-clean bench Ventilate and dry on top for 0.5-1h; use energy of 10-20KeV, dose of 40-150×10 14 ions / cm 2 He + In vacuum (vacuum degree is 2×10 -2 ~5×10 -3 pa), intermittent pulse injection under sterile conditions; then the spores were eluted with phosphate buffer, screened on NBRIY (phytate phosphorus) medium, and after identification, an engineering strain that efficiently decomposes phytate phosphorus was obtained. The ability to decompose phytate phosphorus is increased by 25-35%.
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