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Method for screening strain with high yield of Zhongshengmycin

A technology for high-yielding strains and biotins, applied in microorganism-based methods, biochemical equipment and methods, botanical equipment and methods, etc., can solve problems such as low content of biotins, achieve rational and rapid screening, and wide sources. , the effect of low cost

Pending Publication Date: 2021-02-05
陕西麦可罗生物科技有限公司
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AI Technical Summary

Problems solved by technology

[0005] In order to solve the prior art Streptomyces lilacaceae Hainan variant ( Streptomyces lavendulae var. hainanensis new var. ) lawn is thin, the quality of the lawn decreases after subculture, and the content of Zhongshengmycin is low. The present invention provides a method for screening high-yielding Zhongshengmycin (Zhongshengmycin) strains, which solves the problem of Zhongshengmycin in existing strains. low content technical issues

Method used

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  • Method for screening strain with high yield of Zhongshengmycin
  • Method for screening strain with high yield of Zhongshengmycin
  • Method for screening strain with high yield of Zhongshengmycin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: preparation of original bacterial strain bacterial suspension

[0036] Step 1: Streptomyces lilacinus varietus Hainan var. Streptomyces lavendulae var. hainanensis new var. ) slope (such as figure 1 ) spores were inoculated into the shake flask activation medium at a constant temperature of 30°C and 250rpm for 3 hours to form a germinated bacterial suspension (such as Figure 4 ). Among them, shake flask activation medium ingredients: soluble starch (AR) 1.0%, MgSO 4 ·7H 2 O(AR)0.05%, KH 2 PO 4 (AR) 0.1%, KCl (AR) 0.05%, NaNO 3 (AR) 0.2%, FeSO 4 ·7H 2 O(AR) 0.001%, prepared with distilled water, the pH before disinfection is 6.8-7.2, and the pH after disinfection is controlled at 6.8-7.0.

Embodiment 2

[0037] Embodiment 2: Mutagenesis of Zhongshengmycin Starting Strain

[0038] Step 2 Take 800 μL of the germinated bacterial suspension cultured in Example 1, add 200 μL of 20% glycerol, shake and disperse to prepare the bacterial suspension to be mutated.

[0039] Step 3 Take 15 μL of the bacterial suspension to be mutated and drop it on the slide of the ARTP mutagenesis breeding instrument, spread it evenly, and use the ARTP mutagenesis breeding instrument to undergo ARTP mutagenesis (irradiation power 120W, 25°C, helium flow rate 8-12SLM, irradiation for 70 seconds) to obtain a mutagenic solution.

[0040] Step 4 Wash the mutagenesis solution with sterile saline.

[0041] Step 5 Under red light, dilute the mutagenesis solution with normal saline gradient to 10 -4 、10 -5 、10 -6 , smeared on a plate containing LiCl medium and cultured at a constant temperature of 30°C for 12 days.

[0042] The medium composition of LiCl medium plate: soluble starch (AR) 2.0%, beef extract...

Embodiment 3

[0050] Embodiment 3: Inclined-plane bacterial lawn comparison

[0051] The starting strain 1# and the high-yielding strain 46# were inoculated on the slant medium respectively, and cultured at a constant temperature of 30°C. It was found that the lawn of 46# changed faster than the starting strain 1#. After 12 days of cultivation, 1# bacterial lawn (such as figure 1 ) is thinner, uniform lavender gray overall, while 46# bacterial lawn (such as image 3 ) is thicker than 1#, uniform overall, and the color is darker lavender gray than 1#. Compared with 1#, after two generations of passage, the slant bacterial lawn (such as figure 2 ) becomes thinner and even translucent, uniform and fine, pale pink. After counting the spores on the inclined plane, the number of spores in the test tubes of 1# and 46# were 3.9╳10 9 CFU and 7.9╳10 9 CFU, and the number of spores in the test tube after 1# passage for two generations has dropped to 6.8╳10 8 CFU.

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Abstract

The invention relates to a method for screening a strain with high yield of Zhongshengmycin and aims at solving the technical problem of relatively low content of Zhongshengmycin in an existing strains. The starting strain is Streptomyces lavendulae var.hainanensis new var. The method comprises the following steps: germinating mycelia from the starting strain by shake-flask culture; performing ARTP mutagenesis on the mycelia, performing coating onto a LiCl-contained culture medium panel, and performing culturing; pre-screening the picked single colony blocks with a panel, and transferring thepicked single colonies with relatively large inhibition zone diameters onto a checking slope; and performing shake flask checking on the checking slope, and selecting strains with high content of Zhongshengmycin. According to the method, the strains with high content of Zhongshengmycin are screened, fermentation liquor of the strains has an excellent effect for preventing rice bacterial leaf blight, cabbage soft rot, apple ring rot, apple leaf spot and the like, and can quickly screen out the strains with high content of Zhongshengmycin.

Description

technical field [0001] The invention belongs to the field of microbial pharmacy, and in particular relates to a method for screening high-yield strains of Zhongshengmycin. Background technique [0002] Zhongshengmycin (Zhongshengmycin) belongs to the N-glycoside agricultural antibiotics. It contains 6 effective components, and each component is sequentially increased by a β-lysine. It is active against bacteria, yeast and filamentous fungal diseases. It is mainly used to control rice bacterial blight, cabbage soft rot, apple ring spot and apple leaf spot, etc. It has good control effect, has low toxicity to humans and animals, and does not pollute the environment. Antifungal biopesticide. At present, Zhongshengmycin has been applied and promoted in some provinces and cities across the country, and has achieved certain economic benefits. [0003] Its producing bacteria Streptomyces lavinosa hainanensis ( Streptomyces lavendulae var. hainanensis new var. ), which was isol...

Claims

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Application Information

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IPC IPC(8): C12N15/01C12N13/00C12N1/20C12Q1/04A01N63/28A01N47/18A01P1/00A01P3/00C12R1/56
CPCC12N15/01C12N13/00C12N1/20C12Q1/04A01N63/28A01N47/18C12R2001/56C12N1/205
Inventor 杨玉旺仵林娟潘忠成翁婧李蒲民
Owner 陕西麦可罗生物科技有限公司
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