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High-density fermentation process of rhizobium huakuii

A milk vetch rhizobia, high-density technology, applied in the field of agricultural microorganisms, can solve the problems of long fermentation period and low concentration of fermentation liquid bacteria, and achieve the effect of simple ingredients, simple process and easy operation

Inactive Publication Date: 2012-08-29
HUAZHONG AGRI UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, milk vetch rhizobia has a series of problems such as low bacterial concentration of fermentation liquid and long fermentation cycle. Establish a low-cost and practical technical platform for milk vetch high-density fermentation to promote large-scale milk vetch rhizobia agents. application is important

Method used

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  • High-density fermentation process of rhizobium huakuii

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] A. Cultivation of working seeds Milk Vetch Rhizobium 20026

[0028] From the YM solid medium plate that was streaked and grown for 72 hours, pick a single colony of Milk Vetch Rhizobium 20026 into the liquid seed YM medium, and cultivate it for about 72 hours until the cell concentration is OD 600 When it reaches 0.7~0.8, the working seed solution of fermentation is obtained.

[0029] B. Preparation before inoculation

[0030] (1) Preparation of carbon source mother liquor for feed: Weigh 10 g of glucose and dilute to 1000 mL with deionized water; sterilize under high-pressure steam at 113°C for 25 minutes.

[0031](2) Preparation of nitrogen source mother liquor for feed: Weigh 25g of yeast powder and 25g of peptone to 1000mL with deionized water, and adjust the pH to 7.0-7.2; sterilize at 118°C under high-pressure steam for 25min.

[0032] C. Inoculation

[0033] Add the working seed Milk Vetch Rhizobium 20026 into the fermenter with 50% YM medium at a time accordi...

Embodiment 2

[0041] A. Cultivation of working seeds Milk Vetch Rhizobium 20026

[0042] From the YM solid medium plate that was streaked and grown for 72 hours, pick a single colony of Milk Vetch Rhizobium 20026 into the liquid seed YM medium, and cultivate it for about 72 hours until the cell concentration is OD 600 When it reaches 0.7~0.8, the working seed liquid of fermented milk vetch rhizobium 20026 is obtained.

[0043] B. Preparation before inoculation

[0044] (1) Preparation of carbon source mother liquor for feed: Weigh 15 g of glucose and dilute to 1000 mL with deionized water; sterilize under high-pressure steam at 115°C for 30 min.

[0045] (2) Preparation of nitrogen source mother liquor for feeding: Weigh 40 g of yeast powder and 40 g of peptone and dilute to 1000 mL with deionized water, and adjust the pH to 7.0-7.2; sterilize under high-pressure steam at 121°C for 20 minutes.

[0046] C. Inoculation

[0047] Add the inoculum amount of milk vetch rhizobia 20026 working s...

Embodiment 3

[0054] A. Working seed cultivation

[0055] From the YM solid medium plate that was streaked and grown for 72 hours, pick a single colony of Milk Vetch Rhizobium 20026 into the liquid seed YM medium, and cultivate it for about 72 hours until the cell concentration is OD 600 When it reaches 0.7~0.8, the fermented working seed liquid of Rhizobium vetch 20026 is obtained.

[0056] B. Preparation before inoculation

[0057] (1) Preparation of carbon source mother liquor for feed: Weigh 5 g of glucose and dilute to 1000 mL with deionized water, and sterilize under high-pressure steam at 110° C. for 40 min.

[0058] (2) Preparation of nitrogen source mother liquor for feed: Weigh 10 g of yeast powder and 10 g of peptone and dilute to 1000 mL with deionized water, adjust the pH to 7.0-7.2, and sterilize under high-pressure steam at 115°C for 30 minutes.

[0059] C. Inoculation

[0060] Put the working seed of milk vetch rhizobia 20026 into the fermenter with 40% YM medium at a tim...

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Abstract

The invention belongs to the technical field of agricultural microbiology, and particularly relates to a high-density fermentation process of rhizobium huakuii, which is carried out for obtaining highest biomass, in particular to a fermentation method in which C and N sources different from the original fermentation culture medium are replenished. The biomass of the rhizobium huakuii obtained by the process disclosed by the invention can reach more than 13 billions / mL. According to the process method disclosed by the invention, seeds growing to the late period of logarithm are inoculated intoa YM (Yeast Manitol) culture medium, liquid content in a fermentation tank is 40-60%, the inoculum size is 1-6%, the temperature of the tank is controlled to be 26-30 DEG C, the rotate speed is 400-800rpm, the first time of material replenishment is carried out after 24-48 hours of fermentation, the second time of material replenishment is carried out after 48-72 hours of growing, fermentation iscontinued to 60-84 hours after the second time of material replenishment is completed, and then the fermentation is ended. The replenished material comprises glucose of which the final concentration is 0.5-1.5 g / L, yeast powder of which the final concentration is 1-4g / L and peptone of which the final concentration is 1-4g / L. The process disclosed by the invention is simple, the efficiency is superior to that of the prior art, and the production cost is lower.

Description

technical field [0001] The invention belongs to the technical field of agricultural microorganisms, and in particular relates to a high-density fermentation process of milk vetch rhizobia. Background technique [0002] Milk vetch is a low-cost green manure commonly used in rice crops in southern provinces. In recent years, with the continuous development of agricultural production, the land is often cultivated repeatedly, so the demand for fertilizer sources is also increasing. Vigorously developing milk vetch green manure or bio-organic fertilizer is an effective means to solve this problem. Rhizobium astragula is a beneficial rhizosphere microorganism that symbiotically fixes nitrogen with milk vetch, which can greatly increase the yield of milk vetch. However, in most of the new areas where milk vetch is planted in my country, the root nodule bacteria of milk vetch are generally lacking in the soil, and artificial inoculation must be carried out to ensure the normal grow...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/41
Inventor 陈守文王昌军信健马昕冀志霞李进平祁高富祖秉桥杨树李建平
Owner HUAZHONG AGRI UNIV
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