Bradyrhizobium japonicum capable of effectively fixing nitrogen and culture method and application thereof
A technology of bradyrhizobium and culture method, which is applied in the field of high-efficiency nitrogen-fixing soybean bradyrhizobium, can solve the problems of restricting the industrial production of rhizobia, high fermentation cost, and high price of mannitol, and achieves the simple and easy cultivation method and the source of raw materials Broad, Inexpensive Effects
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Embodiment 1
[0052] Embodiment 1: the cultivation of soybean bradyrhizobium CGMCC No.4346
[0053] The cultivation steps are as follows:
[0054] Prepare slant medium: accurately weigh according to mannitol 10g / L, yeast extract 1.0g / L, dipotassium hydrogen phosphate 0.5g / L, magnesium sulfate 0.5g / L, sodium chloride 0.2g / L, and agar 18g / L. Take mannitol, yeast extract, dipotassium hydrogen phosphate, magnesium sulfate, sodium chloride and agar, and then dissolve it in water. After the dissolution is complete, use acid or alkali to adjust the pH to 6.8-7.0, then add water to reach the above concentration, and mix well Afterwards, it was divided into test tubes, and after autoclaving at a temperature of 121° C. for 30 minutes, the test tubes were placed obliquely while hot to make slant medium.
[0055] Preparation of seed medium: Accurately weigh mannitol, yeast Ointment, dipotassium hydrogen phosphate, magnesium sulfate, sodium chloride, and then dissolved in water, adjusted to pH 6.8-7.0...
Embodiment 2
[0060] Embodiment 2: the cultivation of soybean bradyrhizobium CGMCC No.4346
[0061] Carry out according to the embodiment identical with embodiment 1, just fermentation medium is different. The fermentation medium was prepared according to diethylene glycol 12g / L, yeast extract 3.0g / L, dipotassium hydrogen phosphate 0.5g / L, magnesium sulfate 0.5g / L, sodium chloride 0.2g / L, pH 6.8-7.0. The liquid Bradyrhizobium soybean CGMCC No.4346 inoculum with a viable count of 4.58 billion / ml was obtained.
Embodiment 3
[0062] Embodiment 3: the optimization of soybean bradyrhizobium CGMCC No.4346 fermentation medium
[0063] The low-cost industrial by-products diethylene glycol and sorbitol were selected as carbon sources to replace the carbon source mannitol in the YEM medium, namely the fermentation medium ① and ② described above. The optimization of the fermentation medium was carried out according to the same cultivation method as in Example 1.
[0064] Improved medium of the present invention is compared with traditional YEM medium test result and is listed in Table 1.
[0065] Table 1: Improvement of Bradyrhizobium soybean CGMCC No.4346 medium
[0066]
[0067] Show clearly by the result of table 1, adopt 2 kinds of improved fermentation mediums of the present invention, after cultivating 72h, by dilution plate counting, liquid bradyrhizobium CGMCC No.4346 inoculum viable count can respectively reach 5,030,000,000 / ml and 4.57 billion / ml, higher than the culture results of traditio...
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