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High-efficiency phosphate-solubilizing Clostridium butyricum A5-4 and applications

A technology of Clostridium butyricum and calcium phosphate, applied in the field of agricultural microorganisms, can solve the problems that the research on anaerobic phosphate-dissolving bacteria has not yet been reported, and achieve the effects of wide range of nutritional requirements, promotion of plant growth, and increase of available phosphorus content

Inactive Publication Date: 2011-08-31
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The phosphate solubilizing bacteria currently studied are mainly aerobic bacteria, but the research on anaerobic phosphate solubilizing bacteria has not been reported yet.

Method used

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  • High-efficiency phosphate-solubilizing Clostridium butyricum A5-4 and applications
  • High-efficiency phosphate-solubilizing Clostridium butyricum A5-4 and applications
  • High-efficiency phosphate-solubilizing Clostridium butyricum A5-4 and applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Isolation, screening and identification of Clostridium butyricum A5-4

[0030]Samples were collected from the septic tank bottom mud of Huazhong Agricultural University pig farm in Shizishan District, Hongshan District, Wuhan City, Hubei Province. Bacteria A5-4 was diluted and spread on the improved PVK solid medium plate (see below), cultured anaerobically at 37°C for 7 days, picked the strain with larger ratio of phosphorus-dissolving circle diameter to colony diameter, and Quantitative analysis of its phosphorus dissolving ability. Adopt improved PVK liquid culture medium (no agar is added in the liquid culture medium) under 37 ℃ static culture 7 days, use molybdenum antimony anti-colorimetric method (Zhang Xiangsheng, molybdenum antimony anti-colorimetric method to determine the content of available phosphorus in the phosphorus bacteria fermentation liquid) Analysis of the factors affecting the value, Anhui Agricultural Sciences, 2008, 36 (12) 4822-4823;...

Embodiment 2

[0041] Embodiment 2: Seed culture and fermentation production test 1 of Clostridium butyricum A5-4

[0042] (1) Preparation of Clostridium butyricum A5-4 primary seed liquid: take 200 μL of Clostridium butyricum strain A5-4 preserved in a glycerol tube, inoculate it in a test tube (18×180 mm) containing 15 mL of RCM liquid medium, and incubate at 37° C. Static culture for 10 hours to make Clostridium butyricum A5-4 first-class seed liquid, the number of viable bacteria in the first-class seed liquid reached 3×10 8 a / mL;

[0043] (2) Preparation of secondary seed liquid: the primary seed liquid of step (1) is inserted into the culture bottle equipped with secondary seed medium according to the inoculation amount of 3% by volume, and cultured at 37° C. for 10 hours, Make secondary seed liquid, the number of viable bacteria in the secondary seed liquid reaches 2.5×10 8 a / mL;

[0044] The components and proportions of Clostridium butyricum A5-4 secondary seed medium are as foll...

Embodiment 3

[0049] Embodiment 3: the expanded culture of clostridium butyricum A5-4 and fermentation production test 2

[0050] According to the method of embodiment 1 and embodiment 2, the substratum formulation of secondary seed culture and fermentation production in the present embodiment is as follows:

[0051] Secondary culture medium:

[0052] Glucose 20.0g, corn starch 3g, yeast powder 4.0g, peptone 4.5g, K 2 HPO 4 3g, NaCl 0.3g, MgSO 4 ·7H 2 O0.5g, KCl 0.3g, FeSO 4 ·7H 2 O 0.03g, MnSO 4 ·H 2 O 0.03g, water 1L, pH7.3. Cultivate statically at 37°C for 12 hours, and the number of viable bacteria in the obtained secondary seed liquid reaches 2.8×10 8 a / mL;

[0053] Fermentation medium:

[0054] Glucose 24.0g, corn starch 5.0g soybean meal 38.0g, corn steep liquor 8.0g, peptone 4.0g, K 2 HPO 4 4.0g, NaCl0.3g, MgSO 4 ·7H 2 O 0.5g, FeSO 4 ·7H 2 O 0.02g, MnSO 4 ·H 2 O 0.02g, water 1L, pH7.5. Static culture at 37°C for 60 hours, the number of Clostridium butyricum A5-4...

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Abstract

The invention discloses high-efficiency phosphate-solubilizing Clostridium butyricum, a preparation method and application thereof, belonging to the technical field of agricultural microorganisms. In the invention, one high-efficiency phosphate-solubilizing Clostridium butyricum A5-4 strain is obtained through separation, and the strain is preserved in China Center for Type Culture Collection (CCTCC for short), wherein the preservation number is CCTCC NO: Ma010052. The bacterium is separate from the substrate sludge of a septic tank and has the capacity of efficiently dissolving calcium phosphate and calcium phytate under the anaerobic condition. The bacterium has wide nutrient requirement range, easy cultivation, low cost and storage property and can not cause environmental pollution after being reapplied to the environment. The strain can be developed into a novel microorganism agent applied to aquatic crop cultivation or aquaculture.

Description

technical field [0001] The invention belongs to the technical field of agricultural microorganisms, and specifically relates to a strain of Clostridium butyricum, in particular to the isolation and screening of a strain of Clostridium butyricum that can efficiently dissolve calcium phosphate and calcium phytate under anaerobic conditions, its preparation method and application . Background technique [0002] Phosphorus is crucial to the growth and development of plants, and insufficient supply of phosphorus has gradually become one of the most important factors limiting agricultural output. The total phosphorus content in the soil is about 0.04-0.1%, but only 1.0-2.5% of it can be absorbed and utilized by plants, most of which are accumulated in the soil in an ineffective state (insoluble state), and are difficult to be directly absorbed and utilized by plants. In order to solve the lack of phosphorus and obtain high yields, nearly 30 million tons of phosphorus fertilizers ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C05G3/00C05F11/08C12R1/145
CPCY02W30/40
Inventor 陈守文冀志霞周光林马昕信建柯云
Owner HUAZHONG AGRI UNIV
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