Method for inducing Blakeslea trispora aging strain to rapidly produce a large number of spores

A technology of Bradleyella trispora and aging strains, which is applied in the field of agricultural microorganisms, can solve the problems of difficult separation, purification and optimal breeding of strains, unsuitability for industrial production, weak resistance to miscellaneous bacteria, etc., and achieves cheap raw materials and easy The effect of operation control and sporulation speed

Active Publication Date: 2015-11-04
甘肃汇能生物工程有限公司
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Problems solved by technology

After the aging strains are inserted into the compost, the growth of mycelia is slow, the ability to resist miscellaneous bacteria is weak, and the ability to produce spores is reduced, which brings difficulties to the separation, purification and optimal selection of strains. Therefore, it is urgent to master B. trispora Mass sporulation technology of aged strains
[0004] Zhang Yang, Huazhong University of Science and Technology (Study on the conditions of massive sporulation and fermentation kinetics of B. trispora, 2013,8-21.) in a 250 ml cone Dispense 60 ml of PDA medium in the bottle and inoculate it in the form of spore suspension. The inoculation volume of each Erlenmeyer flask is 1 ml. Cultivate statically on a constant temperature shaker at 20°C for 6 days, and the entire cultivation process lasts 7 days, which is too long and not suitable for industrial production

Method used

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  • Method for inducing Blakeslea trispora aging strain to rapidly produce a large number of spores
  • Method for inducing Blakeslea trispora aging strain to rapidly produce a large number of spores
  • Method for inducing Blakeslea trispora aging strain to rapidly produce a large number of spores

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Aged strain of B. trispora producing large numbers of spores

[0029] (1) Prepare semi-solid medium according to the following formula: malt extract 20g / L, yeast peptone 10g / L, propionate 3g / L, CuSO 4 . 5H 2 O 7 mg / L, FeCl 3 . 6H 2 O 250 mg / L, Co(NO 3 ) 2 . 6H 2 O 4 mg / L, agar powder 18g / L, pH7, plate thickness 1.5mm.

[0030] (2) Inoculate a small amount of mycelium of the aging strain on a semi-solid medium plate under aseptic operation, and culture it at 27°C for 44 h in a dark environment with a relative humidity of 85% and an air unit volume flow rate of 0.5 L / min. , after culturing at 15°C for 16 h, collect and streak the small amount of spores grown on a new sterile plate, and continue culturing under the same culture environment and conditions, and repeat the operation for 5 generations to obtain a large number of trispores Borealis spores. attached figure 1 The number of negative spores before sporulation is 3~7×10 8 pcs / plate; at...

Embodiment 2

[0031] Example 2: Aged strain of B. trispora producing large numbers of spores

[0032] (1) Prepare semi-solid medium according to the following formula: malt extract 5g / L, yeast peptone 5g / L, acetate 20g / L, CuSO 4 . 5H 2 O 1 mg / L, FeCl 3 . 6H 2 O 10 mg / L, Co(NO 3 ) 2 . 6H 2 O 1 mg / L, agar powder 15g / L, pH5.5, plate thickness 1.7mm.

[0033] (2) Inoculate a small amount of mycelium of the aging strain on a semi-solid medium plate under aseptic operation, and culture it at 25°C for 24 h in a dark environment with a relative humidity of 80% and an air unit volume flow rate of 0.5 L / min. , after culturing at 10°C for 12 h, collect and streak the small amount of spores grown on a new sterile plate, and continue culturing under the same culture environment and conditions, and repeat the operation for 5 generations to obtain a large number of trispores Borealis spores. The number of negative bacterial spores before sporulation was detected to be 2~8×10 8 pc / plate, th...

Embodiment 3

[0034] Example 3: Aged strain of B. trispora produces large numbers of spores

[0035] (1) Prepare semi-solid medium according to the following formula: malt extract 40g / L, yeast peptone 20g / L, lactate 1g / L, CuSO 4 . 5H 2 O 10 mg / L, FeCl 3 . 6H 2 O 500 mg / L, Co(NO 3 ) 2 . 6H 2 O 10 mg / L, agar powder 20g / L, pH7.5, plate thickness 1.9mm.

[0036] (2) Inoculate a small amount of mycelium of the aging strain on a semi-solid medium plate under aseptic operation, and culture it at 30°C for 48 h in a dark environment with a relative humidity of 95% and an air unit volume flow rate of 0.5 L / min. , after culturing at 20°C for 24 h, collect and streak the small amount of spores grown on a new sterile plate, and continue culturing under the same culture environment and conditions, and repeat the operation for 5 generations to obtain a large number of trispores Borealis spores. The number of negative bacterial spores before sporulation was detected to be 4~8×10 8 pc / plat...

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Abstract

The present invention relates to a method for inducing a Blakeslea trispora aging strain to rapidly produce a large number of spores. The method comprises: (1) loading a semi-solid culture medium into a culture dish, inoculating the hypha of a Blakeslea trispora aging strain into the semi-solid culture medium, culturing for 24-48 h at a temperature of 25-30 DEG C in a dark environment with a relative humidity of 80-95% and an air flow rate of 0.5 L / min, then culturing for 12-24 h at a temperature of 10-20 DEG C, and growing spores or hyphae on the culture medium flat plate; (2) collecting and streaking the obtained spores or hyphae on another new semi-solid culture medium flat plate, and culturing for the same time as the step (1) in the same environment as the step (1); and (3) repeatedly performing the operations in the step (2), culturing 1-5 generations of the hyphae, and picking or collecting the spores so as to obtain a large number of the spores. According to the present invention, with the method, the spore yield is high, the spore production is rapid, and the problem that no spore or less spores is produced during the Blakeslea trispora (especially negative bacteria) culture process is effectively solved.

Description

technical field [0001] The invention belongs to the technical field of agricultural microorganisms, and relates to a method for inducing rapid and large-scale spore production of aged strains of B. trispora and the application of the obtained spores in the isolation and breeding of B. trispora strains. Background technique [0002] Lycopene is a fat-soluble carotenoid, which has good anti-oxidation and anti-tumor effects, and has certain effects on the prevention of tumors, cardiovascular protection, and maintenance of the integrity of immune cells, so it has attracted the attention of many researchers . The main ways to obtain lycopene are extraction, chemical synthesis and biosynthesis. Among them, biosynthesis is the most promising way to obtain lycopene. Fermentation of lycopene by B. trispora has the advantages of high fermentation unit titer, environmental friendliness, green and natural, and production is not affected by seasons. Therefore, It is widely used in indu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N3/00C12N1/14C12R1/645
Inventor 陈贵才侯保兵聂月美王丽
Owner 甘肃汇能生物工程有限公司
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