Rejuvenation technology of agricultural microorganism strains by using chlorine dioxide

A technology of chlorine dioxide and microorganisms, which is applied in the field of chlorine dioxide rejuvenation of agricultural microbial strains, can solve the problems of cumbersome, heavy workload, and high-level operation requirements, and achieve the effects of short working procedures, good effectiveness, and large screening volume

Active Publication Date: 2009-10-14
JIANGXI LVYUE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the rejuvenation method of pure species isolation can reach the level of "pure colonies" or even "pure cells", the operation requires a high level, and it is extremely cumbersome, and the workload is heavy; rejuvenation through hosts is only feasible for some parasitic bacteria, and it is very Parasi

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0019] Embodiment: 1, the rejuvenation of Coryneform bacteria of glutamicum Coryneform bacteria

[0020] The bacterial strain to be rejuvenated is connected with 50ml culture medium (glucose 2.5%, urea 0.5%, MgSo 4 ·7H 2 O0.04%, K 2 HPO 4 0.1%, pH 6.8-7.07 in a 250ml Erlenmeyer flask (sterilized), cultivated at 30°C and 120r / min for 12h.

[0021] Then add 1000ppm of CLO 2 3.2-5.6ml, treated on a shaker at 30°C and 120r / min for 3min, immediately diluted with sterile water to five times the original volume, oscillated evenly, and smeared on a plate (the medium was glucose 0.1%, beef extract 1.0%, Peptone 0.8%, NaCL 0.5%, agar 2%, pH 7.0-7.2) were incubated at 32°C for 24h.

[0022] The colonies on the plate grew and expanded well, and the microscopic examination showed that the bacteria were robust and uniform.

[0023] Select good bacteria and insert them into shake flasks as strains. After the cultivation is completed, insert them into the fermentation medium (common h...

Embodiment 2

[0024] Example 2: Rejuvenation of the antibiotic Stereptomyces microflavus

[0025] The bacterial strain to be rejuvenated is connected with 50ml of sterilized medium (soy bean cake powder 2%, glucose 2% CaCO 3 0.4%, Nacl 0.3%) in a 250ml Erlenmeyer flask, cultivated at 28°C and 120r / min for 30h.

[0026] Then add 1000ppm of CLO 2 2.1-5.8ml, put on a shaker at 28°C, 120r / min for 3min, immediately dilute to five times the original volume with sterile water, shake evenly, smear on a plate (medium is potato agar), and incubate at 28°C for 36h .

[0027] The growth and expansion of the bacteria on the plate was good, the hyphae under the microscope were thick and branched, and the surface of the colony was pink, with light brown dewdrops and the fragrance of borneol.

[0028] The spores of those with good colonies were drawn into the center line of the plate (potato agar medium), and another 3cm on both sides of the center line, were drawn into cotton wilt bacteria, cultured a...

Embodiment 3

[0029] Embodiment 3: the rejuvenation of potassium-dissolving bacterium Bacillus mucilaginosus

[0030] The strain to be rejuvenated was inserted into 50ml of sterilized medium (soluble starch 0.5%, ammonium sulfate 0.1%, K 2 HPO 4 0.2%, CaCO 3 0.01%, yeast powder 0.02%, 1% ferric chloride trace) in a 250ml Erlenmeyer flask, cultivated at 28°C and 120r / min for 72h.

[0031] Then add 1000ppm of CLO 2 5.4-8.0ml, placed on a shaker, treated at 28°C, 120r / min for 6min, immediately diluted with sterile water to five times the original volume, shaken evenly, spread on a plate (the medium is 1% sucrose, K 2 HPO 4 0.05%, magnesium sulfate 0.02%, Nacl 0.02%, CaCO 3 0.1%, yeast powder 0.04%, agar 2%), cultivated at 28°C for 72h.

[0032] Multiple colorless and transparent mucus-like bacterial lawns formed on the plate, and the bacterial cells were robust under the microscope.

[0033] Insert this rejuvenated thalline into the above-mentioned starch ammonium medium (replace K wi...

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PUM

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Abstract

The invention discloses a rejuvenation technology of agricultural microorganism strains by using chlorine dioxide, which comprises the following steps: firstly, microscopic examination is conducted on a shaking flask culture solution of bacteria to be rejuvenated; when in logarithmic growth phase, adding CLO2 solution so as to cause the concentration of CLO2 in the whole culture solution to reach 40 to 120ppm and diluting with aseptic water after shaking treatment are conducted; bacteria suspension containing remained vigorous thalli is coated on a flat plate for culturing and the remained vigorous thalli is examined and enters a culture medium inclined surface until fully distributing the inclined surface; the specific function detection of the strains is conducted and the strains to be improved in respect of specific function to an expected value enter the culture medium inclined surface for culturing and are put in a fridge for later use. The rejuvenation technology adopts chlorine dioxide as a declining individual eliminating agent, is characterized by being safe, changing no hereditary property of the strains, leaving no potential risk and quite obvious restorative improvement on specific function, and has large screening quantity, short and less working procedures, high effectiveness and quite good operational performance.

Description

technical field [0001] The invention relates to a technology for rejuvenating microorganism strains, in particular to a technology for rejuvenating agricultural microorganism strains with chlorine dioxide. Background technique [0002] The methods to prevent the decline and rejuvenation of strains usually include: 1. Control the number of subcultures; 2. Create good culture conditions; 3. Use different types of cells for inoculation and passage. Generally, the above methods are used to prevent the decline of strains. The methods for rejuvenating bacterial strains include: 1. isolation of pure strains; 2. rejuvenation through the host; 3. elimination of decayed individuals. [0003] Although the above-mentioned methods to prevent the decline of bacterial species have relative effects, they have no absolute effect, because the genetic variation of microorganisms is absolute, while stability is relative. Therefore, in order to ensure high quality and high yield, the strain mus...

Claims

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Application Information

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IPC IPC(8): C12N1/38C12R1/07C12R1/01
Inventor 汪鹤亭吴先道万琼
Owner JIANGXI LVYUE BIO ENG
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