Preparation method of DXQ-1 powder

Abstract

The invention relates to the technical field of microbial agents, discloses a preparation method of DXQ-1 powder, and solves the problems that most of existing microbial agent preparation methods are relatively complex, preparation is often carried out through a complex testing means, and whether the microbial agent has phosphorus dissolving capacity, nitrogen fixing capacity and siderophore generating capacity or not needs to be detected, so that the preparation cost is low. The optimal carrier of the microbial agent is inconvenient to screen. The effect that a worker can conveniently select a carrier of a DXQ-1 microbial agent is achieved, and meanwhile, a reagent is convenient to prepare.

Description

Technical Field

[0001] The invention relates to the technical field of microbial agents, and in particular to a method for preparing DXQ-1 powder. Background Art

[0002] Microbial agents refer to live bacterial preparations made by using porous materials as adsorbents (such as peat and vermiculite) to absorb the fermentation liquid of the target microorganisms after industrial production and propagation. This kind of agent is used for seed dressing or root dipping, and has the functions of directly or indirectly improving soil, restoring soil fertility, preventing soil-borne diseases, maintaining the balance of rhizosphere microbial flora, and degrading toxic substances. The proper use of agricultural microbial agents can increase the yield of agricultural products, improve the quality of agricultural products, reduce the use of chemical fertilizers, reduce costs, improve soil, and protect the ecological environment; Microbial agents are live bacterial preparations made from beneficial microorganisms, and are widely used in feed additives, organic material decomposition, soil improvement and restoration, sewage purification and other fields. For example, BGB microbial agents contain a variety of beneficial microorganisms and trace elements, which can effectively promote the absorption of nutrients by crops, increase fertilizer utilization, improve the microecological environment of crop roots, and increase crop disease resistance and yield; Most of the existing methods for preparing microbial agents are relatively complicated, and often require complex testing methods for preparation. In addition, it is also necessary to test whether the microbial agents have the ability to dissolve phosphorus, fix nitrogen, and produce iron carriers, which makes it inconvenient to screen the most suitable carriers for microbial agents.

[0003] In view of the above problems, a method for preparing DXQ-1 powder is proposed. Summary of the invention

[0004] The purpose of the present invention is to provide a method for preparing DXQ-1 powder, and the device is used to work, thereby solving the problem that most of the existing methods for preparing microbial agents are relatively complicated, often need to be prepared by complex testing means, and also need to detect whether the microbial agents have the ability to dissolve phosphorus, fix nitrogen, and produce iron carriers, which is not convenient for screening the most suitable carrier for the microbial agents.

[0005] To achieve the above object, the present invention provides the following technical solution: a method for preparing DXQ-1 powder, comprising the following steps: Step 1: Add DXQ-1 seed solution to 500 ml of sterilized LB medium at a 1% inoculum volume, and culture at 28 degrees Celsius with a shaking frequency of 180 times per minute; Step 2: Use an electronic balance to weigh nine different carriers of the same mass, and mix them with the DXQ-1 fermentation broth in the same proportion, where the ratio of carrier to fermentation broth is 1:9, and then use a magnetic stirrer to mix the system thoroughly; Step 3: Use an electronic balance to weigh the same mass of heat-resistant protective agent, place the mixed system in a 60-degree Celsius oven to dry, crush the dried sample, dilute the sample after passing through an 80-mesh sieve with sterile water, and then spread it on a plate on LB solid culture medium, count the colonies, and set up three parallels for each group; Step 4: Obtain DXQ-1 powder, wherein the effective viable count is CFU / g, the results of culturing DXQ-1 in modified Martin's medium showed that the number of mold in the inoculum was 0, and the average pH value of DXQ-1 powder was 7.413.

[0006] Preferably, in step 1, the bacterial solution at 0, 12, 24, 36, 48, 60, 72, 84, 96, 108, 120, 132, and 144 hours are taken to measure the absorbance, and a growth curve is drawn to determine the culture time when the number of bacteria is the largest.

[0007] Preferably, the carrier in step 2 is: lignin fiber, bentonite, diatomaceous earth, sodium lignin sulfonate, talcum powder, water, light calcium carbonate, kaolin.

[0008] Preferably, in the step 2, the mixed system is placed in an oven at 60 degrees Celsius for drying, the dried sample is crushed, the sample after passing through an 80-mesh sieve is diluted with sterile water and then coated on a plate with LB solid culture medium, and the colonies are counted, and the fermentation broth is also diluted with sterile water and coated on a plate, and counted, and three parallels are set for each group.

[0009] Preferably, the heat-resistant protective agent in step three is starch, dextrin, sucrose, sodium glutamate, etc., each weighed one gram in mass, starch plus dextrin, starch plus sucrose, starch plus sodium glutamate, dextrin plus sucrose, dextrin plus sodium glutamate, sucrose plus sodium glutamate, etc., and the amount of the heat-resistant protective agent is weighed 0.5 grams respectively.

[0010] Preferably, the optimal vector selected and the DXQ-1 fermentation broth are mixed in the same proportion in the step three, the ratio of the heat-resistant protective agent, the carrier and the fermentation broth is 1:9:90, the system is fully mixed with a magnetic stirrer, the mixed system is placed in an oven at 60 degrees Celsius for drying, the dried sample is crushed, the sample after passing through an 80-mesh sieve is diluted with sterile water and then coated on an LB solid culture medium plate, the colonies are counted, and three parallels are set for each group.

[0011] Preferably, in step three, an electronic balance is used to weigh the heat-resistant protective agent in proportion, and the heat-resistant protective agent is: sucrose plus sodium glutamate: the carrier is light calcium carbonate: the fermentation broth is mixed in a ratio of 1:9:90, 2:8:90, and 3:7:90, and the mixed system is placed in a 60-degree Celsius oven for drying, and the dried sample is crushed. The sample after passing through an 80-mesh sieve is diluted with sterile water and then coated on an LB solid culture medium plate, and the colonies are counted, and three parallels are set for each group.

[0012] Preferably, the pH value measurement method in step 4 is to weigh one gram of sample and put it into a 50-ml beaker, then add 10 ml of distilled water into the beaker and stir evenly, let it stand for 30 minutes, measure the pH of the sample suspension, repeat the reading three times, and calculate the average value.

[0013] Compared with the prior art, the present invention has the following beneficial effects: 1. A DXQ-1 powder preparation method proposed in the present invention solves the problem that most of the existing microbial agent preparation methods are relatively complicated, and often need to be prepared by complex testing means, and also need to detect whether the microbial agent has the ability to dissolve phosphorus, fix nitrogen, and produce iron carriers, which is not convenient for screening the most suitable carrier for the microbial agent. The DXQ-1 seed liquid is added to 500 ml of sterilized LB culture medium at an inoculation rate of 1%, and the culture is shaken at 28 degrees Celsius and 180 rotations per minute. The bacterial liquid at zero, twelve, twenty-four, thirty-six, forty-eight, sixty, seventy-two, eighty-four, ninety-six, one hundred and eight, one hundred and twenty, one hundred and thirty-two, and one hundred and forty-four hours is taken to measure its absorbance, and a growth curve is drawn to determine the culture time with the largest number of bacteria. It can be seen from the results that the longer the culture time, the higher the percentage of spore production rate. Nine different carriers of the same mass are weighed using an electronic balance, and the same proportion is used, wherein the carrier and The ratio of the fermentation broth is 1:9, which can be mixed with the DXQ-1 fermentation broth, and then a magnetic stirrer is used to fully mix the system. By comparing nine different carriers, it is found that the number of colonies in the plate in light calcium carbonate is the largest. It can be seen that the carrier effect of light calcium carbonate is the best. Subsequently, an electronic balance is used to weigh the same mass of heat-resistant protective agent, and it is mixed with the screened optimal carrier and DXQ-1 fermentation broth in the same proportion. A magnetic stirrer is used to fully mix the system. The heat-resistant protective agent, carrier and fermentation broth are 1:9:90. The mixed system is placed in a 60-degree Celsius oven for drying, and the dried sample is crushed. The sample after passing through an 80-mesh sieve is diluted with sterile water and then coated on an LB solid culture medium plate for colony counting. After comparative analysis, it can be seen that among the heat-resistant protective agents, the plate in sucrose plus sodium glutamate has the largest number of colonies. Finally, an electronic balance is used to weigh the heat-resistant protective agent, carrier and fermentation broth in proportions of 1:9:90, 2:8:90, and 3:7:90 and mix them. The mixed system was placed in an oven at 60 degrees Celsius for drying, the dried sample was crushed, the sample after passing through an 80-mesh sieve was diluted with sterile water and then coated on a plate with LB solid culture medium for colony counting. From the data comparison, it can be seen that the ratio of heat-resistant protective agent, carrier and fermentation liquid is 2:8:90, the number of colonies in the plate is the largest, and the number of effective live bacteria is CFU / g, the results of culturing DXQ-1 bacterial agent with modified Martin's medium showed that the number of mold in the agent was 0, and the average pH value of DXQ-1 powder was 7.413, which made it convenient for the staff to select the carrier of DXQ-1 bacterial agent and also facilitated the preparation of reagents. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 This is a schematic diagram of the growth curve of DQX-1 of the present invention; Figure 2 This is a schematic diagram of DXQ-1 culture time screening of the present invention; Figure 3 This is a schematic diagram for selecting the best vector of the present invention; Figure 4 It is a schematic diagram for selecting the best protective agent of the present invention; Figure 5 A schematic diagram for selecting the best ratio of the present invention; Figure 6 This is a schematic diagram of the appearance of the DXQ-1 powder of the present invention. DETAILED DESCRIPTION

[0015] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0016] In order to further understand the content of the present invention, the present invention is described in detail in conjunction with the accompanying drawings.

[0017] Combination Figure 1-Figure 3 , a method for preparing DXQ-1 powder, comprising the following steps: Step 1: Add DXQ-1 seed solution to 500 ml of sterilized LB medium at a 1% inoculum volume, and culture at 28 degrees Celsius with a shaking frequency of 180 times per minute; Step 2: Use an electronic balance to weigh nine different carriers of the same mass, and mix them with the DXQ-1 fermentation broth in the same proportion, where the ratio of carrier to fermentation broth is 1:9, and then use a magnetic stirrer to mix the system thoroughly; Step 3: Use an electronic balance to weigh the same mass of heat-resistant protective agent, place the mixed system in a 60-degree Celsius oven to dry, crush the dried sample, dilute the sample after passing through an 80-mesh sieve with sterile water, and then spread it on a plate on LB solid culture medium, count the colonies, and set up three parallels for each group; Step 4: Obtain DXQ-1 powder, wherein the effective viable count is CFU / g, the results of culturing DXQ-1 in modified Martin's medium showed that the number of mold in the inoculum was 0, and the average pH value of DXQ-1 powder was 7.413.

[0018] In step 1, the bacterial solution at 0, 12, 24, 36, 48, 60, 72, 84, 96, 108, 120, 132, and 144 hours is taken to measure its absorbance, and a growth curve is drawn to determine the culture time when the bacterial cell count is the largest.

[0019] The carrier in step 2 is: lignin fiber, bentonite, diatomaceous earth, sodium lignin sulfonate, talcum powder, water, light calcium carbonate and kaolin.

[0020] In step 2, the mixed system is placed in an oven at 60 degrees Celsius for drying, the dried sample is crushed, the sample after passing through an 80-mesh sieve is diluted with sterile water and then spread on a plate on LB solid culture medium for colony counting. The fermentation liquid is also diluted with sterile water and spread on a plate for counting. Three parallels are set for each group.

[0021] The heat-resistant protective agents in step three are starch, dextrin, sucrose, sodium glutamate, etc., each weighed one gram in mass, and combinations of starch plus dextrin, starch plus sucrose, starch plus sodium glutamate, dextrin plus sucrose, dextrin plus sodium glutamate, sucrose plus sodium glutamate, etc., and the amount of the heat-resistant protective agents is weighed 0.5 grams respectively.

[0022] In step three, the optimal vector selected and the DXQ-1 fermentation broth were mixed in the same proportion, the ratio of the heat-resistant protective agent, the carrier, and the fermentation broth was 1:9:90, and the system was fully mixed with a magnetic stirrer. The mixed system was placed in an oven at 60 degrees Celsius for drying, and the dried sample was crushed. The sample after passing through an 80-mesh sieve was diluted with sterile water and then coated on an LB solid culture medium plate. The colonies were counted, and three parallel plates were set up for each group.

[0023] Step three: Use an electronic balance to weigh the heat-resistant protective agent in proportion. The heat-resistant protective agent is: sucrose plus sodium glutamate: carrier is light calcium carbonate: fermentation broth is mixed in the ratio of 1:9:90, 2:8:90, and 3:7:90. The mixed system is placed in a 60-degree Celsius oven to dry, and the dried sample is crushed. The sample after passing through an 80-mesh sieve is diluted with sterile water and then coated on an LB solid culture medium plate, and the colonies are counted. Three parallels are set up for each group.

[0024] The pH value measurement method in step 4 is to weigh one gram of sample and put it into a 50 ml beaker, then add 10 ml of distilled water into the beaker and stir evenly, let it stand for 30 minutes, measure the pH of the sample suspension, repeat the reading three times, and calculate the average value.

[0025] The present invention will be further described below in conjunction with the embodiments. Example

[0026] In order to solve the problem that most of the existing microbial agent preparation methods are relatively complicated, often need to be prepared by complex testing means, and also need to detect whether the microbial agent has the ability to dissolve phosphorus, fix nitrogen, and produce iron carriers, which is not convenient for screening the most suitable carrier for the microbial agent, this embodiment discloses the following technical solutions, specifically as follows Figure 1-Figure 6As shown, DXQ-1 seed solution was added to 500 ml of sterilized LB medium at a 1% inoculation rate, and cultured at 28 degrees Celsius with 180 rotations per minute. The bacterial solution was taken at 0, 12, 24, 36, 48, 60, 72, 84, 96, 108, 120, 132, and 144 hours to measure its absorbance, and a growth curve was drawn to determine the culture time with the largest number of bacteria. From the results, it can be seen that the longer the culture time, the higher the percentage of spore production rate. Nine different carriers of the same mass were weighed using an electronic balance and mixed with DXQ-1 fermentation solution at the same ratio, wherein the ratio of carrier to fermentation solution was 1:9, and then the system was fully mixed using a magnetic stirrer. By comparing the nine different carriers, it was found that The number of colonies in the plate containing light calcium carbonate is the largest, which shows that the carrier effect of light calcium carbonate is the best. Subsequently, an electronic balance is used to weigh the same mass of heat-resistant protective agent, and it is mixed with the selected optimal carrier and DXQ-1 fermentation broth in the same proportion. A magnetic stirrer is used to fully mix the system. The heat-resistant protective agent, carrier and fermentation broth are 1:9:90. The mixed system is placed in a 60-degree Celsius oven to dry, and the dried sample is crushed. The sample that has passed through an 80-mesh sieve is diluted with sterile water and then coated on an LB solid culture medium plate. The colonies are counted. After comparative analysis, it can be seen that among the heat-resistant protective agents, the plate containing sucrose plus sodium glutamate has the largest number of colonies. Finally, an electronic balance is used to weigh the heat-resistant protective agent, carrier and fermentation broth in proportions of 1:9:90, 2:8:90, and 3:7:90 and mix them. The mixed system was placed in an oven at 60 degrees Celsius for drying, the dried sample was crushed, the sample after passing through an 80-mesh sieve was diluted with sterile water and then coated on a plate with LB solid culture medium for colony counting. From the data comparison, it can be seen that the ratio of heat-resistant protective agent, carrier and fermentation liquid is 2:8:90, the number of colonies in the plate is the largest, and the number of effective live bacteria is CFU / g, the results of culturing DXQ-1 bacterial agent with modified Martin's medium showed that the number of mold in the agent was 0, and the average pH value of DXQ-1 powder was 7.413, which made it convenient for the staff to select the carrier of DXQ-1 bacterial agent and also facilitated the preparation of reagents.

[0027] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.

[0028] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing DXQ-1 powder, characterized in that: The steps include: Step 1: Add DXQ-1 seed solution to 500 ml of sterilized LB medium at a 1% inoculum volume, and culture at 28 degrees Celsius with a shaking frequency of 180 times per minute; Step 2: Use an electronic balance to weigh nine different carriers of the same mass, and mix them with the DXQ-1 fermentation broth in the same proportion, where the ratio of carrier to fermentation broth is 1:9, and then use a magnetic stirrer to mix the system thoroughly; Step 3: Use an electronic balance to weigh the same mass of heat-resistant protective agent, place the mixed system in a 60-degree Celsius oven to dry, crush the dried sample, dilute the sample after passing through an 80-mesh sieve with sterile water, and then spread it on a plate on LB solid culture medium, count the colonies, and set up three parallels for each group; Step 4: Obtain DXQ-1 powder, wherein the effective viable count is CFU / g, the results of culturing DXQ-1 in modified Martin's medium showed that the number of mold in the inoculum was 0, and the average pH value of DXQ-1 powder was 7.

413.

2. A method for preparing DXQ-1 powder according to claim 1, characterized in that: In the step 1, the bacterial solution at 0, 12, 24, 36, 48, 60, 72, 84, 96, 108, 120, 132 and 144 hours is taken to measure its absorbance, and a growth curve is drawn to determine the culture time when the bacterial cell number is the largest.

3. A method for preparing DXQ-1 powder according to claim 1, characterized in that: The carrier in step 2 is: lignin fiber, bentonite, diatomaceous earth, sodium lignin sulfonate, talcum powder, water, light calcium carbonate, and kaolin.

4. A method for preparing DXQ-1 powder according to claim 1, characterized in that: In the step 2, the mixed system is placed in an oven at 60 degrees Celsius for drying, the dried sample is crushed, the sample after passing through an 80-mesh sieve is diluted with sterile water and then coated on a plate with LB solid culture medium, and the colonies are counted. The fermentation liquid is also diluted with sterile water and coated on a plate, and counted. Three parallels are set for each group.

5. The method for preparing DXQ-1 powder according to claim 1, characterized in that: The heat-resistant protective agents in the step three are starch, dextrin, sucrose, sodium glutamate, etc., each weighed one gram in weight, and combinations of starch plus dextrin, starch plus sucrose, starch plus sodium glutamate, dextrin plus sucrose, dextrin plus sodium glutamate, sucrose plus sodium glutamate, etc., and the amount of the heat-resistant protective agents is weighed 0.5 grams respectively.

6. The method for preparing DXQ-1 powder according to claim 1, characterized in that: In the step three, the optimal carrier screened out and the DXQ-1 fermentation broth are mixed in the same proportion, the ratio of the heat-resistant protective agent, the carrier, and the fermentation broth is 1:9:90, and the system is fully mixed using a magnetic stirrer. The mixed system is placed in an oven at 60 degrees Celsius for drying, and the dried sample is crushed. The sample after passing through an 80-mesh sieve is diluted with sterile water and then coated on an LB solid culture medium plate, and the colonies are counted. Three parallels are set for each group.

7. The method for preparing DXQ-1 powder according to claim 1, characterized in that: In the step three, an electronic balance is used to weigh the heat-resistant protective agent in proportion. The heat-resistant protective agent is: sucrose plus sodium glutamate: the carrier is light calcium carbonate: the fermentation liquid is mixed in a ratio of 1:9:90, 2:8:90, and 3:7:

90. The mixed system is placed in a 60-degree Celsius oven for drying, and the dried sample is crushed. The sample after passing through an 80-mesh sieve is diluted with sterile water and then coated on an LB solid culture medium plate, and the colonies are counted. Three parallel plates are set for each group.

8. The method for preparing DXQ-1 powder according to claim 1, characterized in that: The pH value measurement method in step 4 is to weigh one gram of sample and put it into a 50 ml beaker, then add 10 ml of distilled water into the beaker and stir evenly, let it stand for 30 minutes, measure the pH of the sample suspension, repeat the reading three times, and calculate the average value.

Images