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Rhizobium astragula capable of effectively fixing nitrogen and culture method and application thereof

The technology of a rhizobium rhizobia and a culturing method is applied in the field of high-efficiency nitrogen-fixing rhizobium rhizobia. Inexpensive effect

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 components mannitol and yeast extract is relatively high, which directly causes the high cost of fermentation, thus restricting the industrial production of rhizobia

Method used

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  • Rhizobium astragula capable of effectively fixing nitrogen and culture method and application thereof
  • Rhizobium astragula capable of effectively fixing nitrogen and culture method and application thereof
  • Rhizobium astragula capable of effectively fixing nitrogen and culture method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1: the cultivation of milk vetch rhizobia CGMCC No.4347

[0064] The cultivation steps are as follows:

[0065] 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.

[0066] 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 wi...

Embodiment 2

[0071] Embodiment 2: Optimizing the fermentation medium of milk vetch rhizobia CGMCC No.4347

[0072] Choose cheap industrial by-product soybean molasses as carbon source to replace part of carbon source mannitol in YEM medium; choose sodium glutamate as nitrogen source to replace part of nitrogen source yeast extract in YEM medium. The optimization of the fermentation medium was carried out in the same culture method as in Example 1.

[0073] Among them, soybean molasses needs to be hydrolyzed first. The specific method is: dilute concentrated sulfuric acid (18.4mol / L) with water to 1mol / L, then mix equal volumes of soybean molasses with a solid content of 60% and diluted 1mol / L sulfuric acid, and sterilize at a temperature of 121°C. hydrolysis.

[0074] Table 1 shows the test results when only the carbon source is changed without changing other components in the existing YEM medium.

[0075] Table 1: CGMCC No.4347 medium for rhizobia CGMCC No.4347

[0076] Carbon source ...

Embodiment 3

[0087] Embodiment 3: the preparation of solid milk vetch rhizobium CGMCC No.4347 bacterial agent

[0088] Peat treatment: Dry the peat naturally, then crush it, pass through a 100-mesh sieve, adjust the pH to 7.0 with quicklime powder, and sterilize at 121°C for 1 hour.

[0089] The acquisition of the milk vetch rhizobium CGMCC No.4347 microbial agent containing peat: the liquid milk vetch rhizobium CGMCC No.4347 bacterial agent obtained in Example 1 and the treated sterilized, dry, pH 7.0~7.2 The peat was mixed according to the weight ratio of 1:2.5, and then multiplied and cultured at 28° C. for 24 hours to obtain solid milk vetch rhizobium CGMCC No.4347 inoculum, and the number of viable bacteria was more than 500 million / g.

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Abstract

The invention relates to rhizobium astragula capable of effectively fixing nitrogen and a culture method and application thereof. The rhizobium astragula CGMCC No.4347 is cultured by the steps of strain activation, seed culture, fermentation culture and the like. The invention has the following beneficial effects: when the fermentation culture medium improved by the invention is adopted for culturing, the viable count of the fermentation liquor is far higher than that in the prior art; compared with the bacteria which are not inoculated on astragalus sinicus, the rhizobium astragula agent can ensure the fresh weight of the astragalus sinicus plants to be increased by more than 241%, the nitrogen content of the plants to be increased by more than 86% and the yield to be increased by more than 47% after being inoculated on the astragalus sinicus; compared with the existing produced strains, the rhizobium astragula agent ensures the fresh weight of the astragalus sinicus plants to be increased by more than 58%, the nitrogen content of the plants to be increased by more than 18% and the yield to be increased by more than 12%, thus being an excellent strain for production of the rhizobium astragula and inoculation of the astragalus sinicus; 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 milk vetch rhizobia, and also relates to the cultivation method of the rhizobia and its use. 【Background technique】 [0002] Milk vetch is a perennial herbaceous plant of the genus Astragalus in the leguminous family. It has high green yield, good fertilizer quality, high nutrient content, easy to decompose, high fertilizer efficiency, and good persistence. It is a high-quality and high-yield organic fertilizer. Milk vetch can greatly increase soil organic matter, enhance soil water retention, fertilizer retention, and heat preservation performance. Milk vetch is usually inter-sowed in late rice fields in autumn and used as base fertilizer for early rice, which can solve the fertilizer problem of double-cropping rice. It is the most important winter green manure crop in rice fields in my country. [0003] R...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/02A01P21/00C12R1/41
Inventor 杜迎辉高玉爽刘书君苏海榆徐志文
Owner 领先生物农业股份有限公司
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