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

Active Publication Date: 2011-09-07
领先生物农业股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the price of the main component mannitol is relatively high, which directly causes the high cost of its fermentation, thus restricting the industrial production of rhizobia

Method used

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  • Bradyrhizobium japonicum capable of effectively fixing nitrogen and culture method and application thereof
  • Bradyrhizobium japonicum capable of effectively fixing nitrogen and culture method and application thereof
  • Bradyrhizobium japonicum capable of effectively fixing nitrogen and culture method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention relates to bradyrhizobium japonicum capable of effectively fixing nitrogen and a culture method and application thereof. The bradyrhizobium japonicum CGMCC No.4346 is cultured by the steps of strain activation, seed culture, fermentation culture and the like. The invention has the following beneficial effects: sorbitol and diethylene glycol are utilized to replace the carbon source, mannitol in the traditional YEM (yeast extract mannitol) culture medium, and when the improved fermentation culture medium is adopted for culturing, the viable count of the fermentation liquor is higher than that of the YEM culture medium; compared with the rhizobium inoculants which are not inoculated on the soy beans, the bradyrhizobium japonicum agent ensures the theoretical yield of the soy beans to be increased by 16.7-17.2%, the crude fat content to be increased by 0.65-1.21% and the protein content to be increased by 1.29-1.60% after being inoculated on the soy beans, thus being an excellent strain for production of rhizobium japonicum and inoculation of the soy beans; and the culture method is simple and practical, and the raw materials have wide sources and low price and are suitable for large-scale industrialized production.

Description

【Technical field】 [0001] The invention belongs to the technical field of rhizobia. More specifically, the present invention relates to a high-efficiency nitrogen-fixing soybean bradyrhizobium, and also relates to a cultivation method of the rhizobia and its use. 【Background technique】 [0002] Inoculation of rhizobia is an internationally recognized biological nitrogen fixation technology, which has been adopted by advanced agricultural countries. Soybean cultivation has caused great pollution to the environment due to the long-term application of large amounts of chemical fertilizers, especially nitrogen fertilizers. Soybean rhizobia inoculation can reduce the amount of nitrogen fertilizer used during soybean planting, promote nodulation of soybean roots, enhance symbiotic nitrogen fixation, and increase soybean yield. protein content. At present, all parts of our country have attached great importance to the application of soybean rhizobia inoculation technology. [000...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/02A01P21/00C12R1/01
Inventor 杜迎辉苏海榆高玉爽王琰李春艳杜月芳徐志文
Owner 领先生物农业股份有限公司
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