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Preparation method of biocontrol bacterium immobilized microspheres optimized by response surface methodology and calculation method of embedding rate of biocontrol bacterium immobilized microspheres

A technology of response surface method and calculation method, applied in biochemical equipment and methods, methods based on microorganisms, fixed on/in organic carriers, etc., can solve the problems of difficult colonization, inconspicuous antagonism, and short action time And other issues

Pending Publication Date: 2021-08-13
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in practical applications, direct spraying of free biocontrol bacteria on plants will cause problems such as short action time, difficulty in colonization, and insignificant antagonism, which will greatly reduce its biocontrol efficiency.

Method used

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  • Preparation method of biocontrol bacterium immobilized microspheres optimized by response surface methodology and calculation method of embedding rate of biocontrol bacterium immobilized microspheres
  • Preparation method of biocontrol bacterium immobilized microspheres optimized by response surface methodology and calculation method of embedding rate of biocontrol bacterium immobilized microspheres
  • Preparation method of biocontrol bacterium immobilized microspheres optimized by response surface methodology and calculation method of embedding rate of biocontrol bacterium immobilized microspheres

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Experimental program
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Effect test

specific Embodiment approach 1

[0053] A method for preparing immobilized microspheres of biocontrol bacteria optimized by response surface methodology, comprising the following steps:

[0054] Step 1. Transplant the Glioma pink strain into PDA culture under aseptic conditions, place it in a constant temperature incubator for purification and culture, transfer the strain after purification into PDB liquid medium with antibiotics, and place it in a constant temperature shaker After vibrating cultivation in the bed, Glycium pinkis fermented liquid is obtained, stand-by;

[0055] Step 2. After sterilizing the sodium alginate solution with a concentration of 3wt%, add the Gliocladium rosea fermentation liquid obtained in Step 1 and stir and mix to obtain a Glioma rosea mixed solution for use;

[0056] Step 3, slowly drop the 2.5wt% sterilized CaCl with the concentration of 2.5wt% of the sterilized CaCl mixed solution that step 2 obtains 2 In the solution, after hardening, immobilized microspheres of Gliocladium...

specific Embodiment approach 2

[0069] The method for calculating the embedding rate of the biocontrol bacteria immobilized microspheres optimized by the response surface method prepared according to a method comprises the following steps:

[0070] Step a, prepare Na 2 HPO 4 Mixed solution with citric acid, adjust the pH value, sterilize at 121°C for 15 minutes, and use it as a solution for unpacking;

[0071] Step b. Put the biocontrol bacteria-immobilized microspheres into the uncapsulation solution prepared in step a, so that the biocontrol bacteria-immobilized microspheres are completely dissolved in the uncapsulation solution, and then take the dissolved uncapsulation solution and spread it on the surface of the petri dish for culture , and counted by plate counting method after bacterial colonies grow out;

[0072] Step c, calculating the embedding rate by plate counting method.

[0073] A method for calculating the embedding rate of biocontrol bacteria immobilized microspheres optimized by response...

specific Embodiment approach 3

[0078] A method for preparing immobilized microspheres of biocontrol bacteria optimized by response surface methodology, comprising the following steps:

[0079] Step 1. Transplant the Glioma pink strain into PDA culture under aseptic conditions, place it in a constant temperature incubator for purification and culture, transfer the strain after purification into PDB liquid medium with antibiotics, and place it in a constant temperature shaker After vibrating cultivation in the bed, Glycium pinkis fermented liquid is obtained, stand-by;

[0080] Step 2. After sterilizing the sodium alginate solution, add the Gliocladium pink fermentation liquid obtained in Step 1 and stir and mix to obtain a mixed solution of Glioma pink, which is set aside;

[0081] Step 3. Slowly drip the G. pink mixed solution obtained in step 2 into sterilized CaCl 2 In the solution, after hardening, immobilized microspheres of Gliocladium rosea are formed, wherein the dosage of Glioma rosea fermentation ...

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Abstract

The invention discloses a preparation method of biocontrol bacterium immobilized microspheres optimized by a response surface methodology, and belongs to the field of biocontrol bacterium agents. The invention aims to optimize the preparation process of the immobilized bacterium microspheres, and finds the optimal process conditions based on the response surface methodology optimization, so that the effectiveness of the immobilized bacterium microspheres in practical application is improved. The method comprises the following steps of transferring a Clonostachys rosea strain into PDA culture under a sterile condition, transferring the strain subjected to purification culture into a PDB liquid culture medium added with antibiotics, placing the PDB liquid culture medium in a constant-temperature shaking table for shaking culture to obtain Clonostachys rosea fermentation liquor, adding a sodium alginate solution, slowly dropwise adding into a sterilized CaCl2 solution by using an injector, and performing hardening to form the Clonostachys rosea immobilized microspheres. The influence of different conditions on the Clonostachys rosea embedding rate in the embedding process is explored by utilizing the response surface methodology, a continuous variable curved surface model is established, and the optimal embedding condition is fitted and determined through a regression equation, so that the maximum efficacy of microorganisms is exerted.

Description

technical field [0001] The invention belongs to the field of biocontrol bacteria agents; in particular, it relates to a preparation method of biocontrol bacteria immobilized microspheres optimized by response surface method and a calculation method of embedding rate. Background technique [0002] As a new type of pest control method, plant biological control has been widely used. Because of its many advantages such as no drug resistance, being friendly to human health and the ecological environment, it has become one of the approaches with great potential for research and development in pest control. Glioma pink, as a member of the genus Glioma, can effectively control botrytis cinerea, leaf mold and other serious diseases, and is one of the broad-spectrum disease-resistant biocontrol fungi with great potential. However, in practical applications, direct spraying of free biocontrol bacteria on plants will cause problems such as short action time, difficulty in colonization, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/10C12N11/04C12N1/14C12Q1/06A01P3/00C12R1/645
CPCC12N11/10C12N11/04C12N1/14C12Q1/06
Inventor 王傲雪李鑫茂陈旭安丽东刘佳音陈秀玲张瑶仇有文冯明芳
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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