CN114934039B - Non-spore microbial agent with prolonged shelf life and preparation method thereof - Google Patents
Non-spore microbial agent with prolonged shelf life and preparation method thereof Download PDFInfo
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 116
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 230000002035 prolonged effect Effects 0.000 title description 5
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- 239000007788 liquid Substances 0.000 claims abstract description 80
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- 230000001580 bacterial effect Effects 0.000 claims abstract description 44
- 241000589614 Pseudomonas stutzeri Species 0.000 claims abstract description 34
- 241000894006 Bacteria Species 0.000 claims abstract description 32
- 241000827905 Pantoea alhagi Species 0.000 claims abstract description 18
- 241000520272 Pantoea Species 0.000 claims abstract description 8
- 238000001035 drying Methods 0.000 claims description 23
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Abstract
The invention discloses a non-spore microbial agent for prolonging the shelf life and a preparation method thereof, wherein the non-spore microbial agent comprises non-spore microbial liquid and turfy soil; the non-spore microbial bacterial liquid is bacterial liquid of any one or more of the following bacteria of the Pantoea alhagi, the Pseudomonas stutzeri and other non-spore bacteria. Compared with the general commercial liquid type bacillus, the powder type non-spore microbial agent provided by the invention has the advantages of greatly improved normal temperature preservation stability and long-acting property, environment friendliness, no pollution and convenience in storage. Wherein, the preservation effect of the Pantoea camelina XK-11 microbial inoculum is best, the shelf life is 180d, and the preservation effect is effectively improved by 35 times; secondly, the pseudomonas stutzeri NRCB010 microbial inoculum has a shelf life of 360d, and is effectively improved by 11 times.
Description
Technical Field
The invention relates to the field of microbial agents, in particular to a non-spore microbial agent with a prolonged shelf life and a preparation method thereof.
Background
In the new era of ecological agriculture, the agricultural fertilization technology tends to be green ecological, and excessive application of common chemical fertilizers is unfavorable for maintaining ecological balance and protecting environment. The microbial agent has the characteristics of long-acting non-toxicity, no pollution, energy conservation, low cost and the like, can be divided into a liquid microbial agent and a solid microbial agent according to the appearance form of the microbial agent, and has the advantages of high production efficiency, short culture period and the like, but also has the defects of inconvenient transportation, short preservation time and the like; compared with the liquid microbial agent, the solid microbial agent has the advantages of convenient storage and transportation, high microbial activity, long preservation period and the like. At present, most of commercial microbial agents are liquid bacillus, and preparation and preservation of non-spore gram-negative microbial agents still have a plurality of barriers, mainly because the microbial agents have low survival rate and poor preservation stability, so that the exploration of a suitable and low-cost microbial agent preparation method is quite practical.
At present, no very mature keep-alive method exists for the non-spore microbial inoculum, the shelf life of the microbial inoculum can be prolonged to a certain extent by a spray drying technology, but the high temperature condition in the drying process has a great influence on the survival rate of cells. Therefore, the invention provides a powdery non-spore microbial agent for normal temperature preservation and a preparation method thereof.
Disclosure of Invention
The invention aims to: the invention aims to solve the technical problem of insufficient preservation stability of a commercial liquid type bacillus agent, and provides application of turfy soil in prolonging the shelf life of a non-spore microbial agent so as to solve two key problems of low survival rate and poor preservation stability of the liquid type bacillus agent.
The invention also solves the technical problem of providing a preparation method of the non-spore microbial agent.
The invention is characterized in that: the method has the advantages that the moisture content of the microbial inoculum dry powder is reduced to the maximum extent, and meanwhile, the cell death is reduced, so that the core break of the problem is solved, and the low-cost biomass carrier has the effects of promoting cell adhesion, proliferation and nutrition supply and has good application prospects in the development of non-spore microbial inoculum products. The turfy soil is a product in the development process of the swamps, is a pure natural, pollution-free and pollution-free green substance, and has very wide application. The turfy soil has high organic matter content and good biodegradability, contains a large amount of humus and a part of mineral substances, is used as a recyclable biomass material, and provides a good carrier for adsorbing and preserving microbial agents. The turfy soil is soft in texture, and the produced fertilizer has loose appearance, and is convenient and quick in production, storage and use; humic acid contained in the turfy soil has adsorption effect, can increase the aggregate structure of the soil, can be used for improving the soil in water and dry fields, obviously improves the fertilizer strength, has low cost, and greatly improves the utilization rate and long-acting property of the microbial agent.
In order to solve the first technical problem, the invention discloses application of turfy soil in prolonging the shelf life of a non-spore microbial agent.
Wherein the non-spore microbial agent comprises non-spore microbial liquid and turfy soil; the non-spore microbial bacterial liquid is bacterial liquid of any one or more of the following bacteria of the Pantoea alhagi, the Pseudomonas stutzeri and other non-spore bacteria.
In some embodiments, the Pantoea alhagi and Pseudomonas stutzeri are Pantoea alhagi XK-11 and Pseudomonas stutzeri NRCB010, respectively, which are all existing bacteria.
Wherein, in the non-spore microbial agent, the mass contents of the non-spore microbial liquid and the turfy soil are respectively 20-50% and 50-80%.
Wherein the viable count of the non-spore microbial agent is 70-100 hundred million/g.
Wherein, when the non-spore microbial liquid is the liquid of the Pantoea alhagi, the viable count of the non-spore microbial agent is more than 9.6 hundred million/g after 180 days.
Wherein, when the non-spore microbial liquid is pseudomonas stutzeri bacterial liquid, the viable count of the non-spore microbial agent is more than 8.4 hundred million/g after 180 days.
In order to solve the second technical problem, the invention discloses a preparation method of the non-spore microbial agent, which comprises the following steps:
s1: culturing bacterial liquid: respectively carrying out aerobic culture on any one or more bacteria of the alhagi, the pseudomonas stutzeri and other non-spore bacteria to obtain non-spore microbial bacteria liquid;
s2: bacterial liquid adsorption: uniformly stirring the non-spore microbial liquid obtained in the step S1 with turfy soil, namely adsorbing with the turfy soil to obtain an undried wet non-spore microbial agent;
s3: and (3) microbial inoculum drying: and (3) drying the undried wet non-spore microbial agent obtained in the step (S2) to obtain the non-spore microbial agent.
In the step S1, the mass ratio of the non-spore microorganism bacterial liquid to the turfy soil is (0.5-3) 1, preferably (1-2) 1, and more preferably 2:1, 3:2, and 4:3.
In the step S1, the viable count in the non-spore microorganism bacterial liquid is 20-40 hundred million/mL.
In the step S3, the drying is carried out in an oven with the temperature of 20-40 ℃ for 12-24 hours.
The commercial microbial agent mainly comprises a liquid microbial agent and a solid microbial agent, and compared with the liquid microbial agent, the solid microbial agent has the advantages of convenience in storage and transportation, high stability, high microbial activity, long preservation period and the like. The powdery microbial agent obtained by the immobilized microbial technology has the advantages of lasting and remarkable effect, green and pollution-free effects and the like when applied to agriculture. The turfy soil is a pure natural, pollution-free and pollution-free green substance, is used as a biomass material capable of being recycled, contains a large amount of organic matters and humus, and can provide a good carrier for adsorbing and preserving microbial agents. The prepared powdery non-spore agricultural microbial agent is low in cost, has loose appearance, is convenient and quick in production, storage and use, and greatly improves the normal-temperature preservation stability and long-acting property of the non-spore microbial agent.
The beneficial effects are that: compared with the prior art, the invention has the following advantages:
the invention uses the turfy soil as the adsorption matrix, mixes the turfy soil with the non-spore microbial liquid uniformly and dries the turfy soil to prepare the powdery non-spore microbial agent for normal temperature preservation. Wherein, the preservation effect of the Pantoea camelina XK-11 microbial inoculum is best, the shelf life is 180d, and the preservation effect is effectively improved by 35 times; secondly, the pseudomonas stutzeri NRCB010 microbial inoculum has a shelf life of 360d, and is effectively improved by 11 times.
Drawings
The foregoing and/or other advantages of the invention will become more apparent from the following detailed description of the invention when taken in conjunction with the accompanying drawings and detailed description.
FIG. 1 is an external view of a prepared powdery non-spore microbial agent product.
FIG. 2 is a scanning electron microscope image of the front and rear of the turfy soil loaded Albizia camelina XK-11 (example 3).
FIG. 3 is a scanning electron microscope image of Pseudomonas stutzeri NRCB010 loaded with turfy soil (example 5).
FIG. 4 shows the elemental composition of turfy soil obtained by SEM-EDS analysis.
FIG. 5 is a graph showing the colony formation of Pantoea alhagi XK-11 in a culture medium of Pantoea alhagi under normal temperature preservation at 25℃with time (example 3).
FIG. 6 is a graph showing the colony formation in the Pseudomonas stutzeri NRCB010 strain agent over time under the condition of normal temperature preservation at 25 ℃ (example 5).
Detailed Description
The experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagents and materials, unless otherwise specified, are commercially available.
The strain preservation number of the alhagi pantoea XK-11 in the following examples is CGMCC No.15525 and the strain preservation number of the Pseudomonas stutzeri NRCB010 is CGMCC No.19067.
In the following examples, the mass ratio of the non-spore microorganism bacterial liquid to the turfy soil in the step (2) is as follows.
Example 1
(1) Culturing bacterial liquid: culturing the strain liquid by adopting the alhagi pantoea XK-11, wherein the culture conditions are as follows: taking LB liquid culture, sterilizing at 121 ℃ for 20min in a high-temperature sterilizing pot, taking out the strain stored at-80 ℃ in advance to be completely dissolved, inoculating the strain into a sterile LB liquid culture medium, and culturing for 12h in a shaking table at 37 ℃ to obtain an activated strain for standby. Calculating the number of viable bacteria by adopting a dilution coating method to obtain non-spore microbial liquid with the initial effective viable bacteria number of 20 hundred million/ml;
(2) Bacterial liquid adsorption: adsorbing the non-spore microorganism bacterial liquid obtained in the step (1) by turfy soil according to the following steps of 2:1, uniformly mixing to obtain an undried wet non-spore microbial agent;
(3) And (3) microbial inoculum drying: and (3) putting the non-spore microbial agent obtained in the step (2) into a baking oven with the temperature of 20 ℃ for drying for 12 hours, and obtaining the powdery non-spore microbial agent with the initial effective viable count of 70 hundred million/g.
Example 2
(1) Culturing bacterial liquid: culturing the strain liquid by adopting the alhagi pantoea XK-11, wherein the culture conditions are as follows: taking LB liquid culture, sterilizing at 121 ℃ for 20min in a high-temperature sterilizing pot, taking out the strain stored at-80 ℃ in advance to be completely dissolved, inoculating the strain into a sterile LB liquid culture medium, and culturing in a shaking table at 37 ℃ for 18h to obtain an activated strain for later use. Calculating the number of viable bacteria by adopting a dilution coating method to obtain non-spore microbial liquid with the initial effective viable bacteria number of 30 hundred million/ml;
(2) Bacterial liquid adsorption: adsorbing the non-spore microorganism bacterial liquid obtained in the step (1) by turfy soil according to the following steps of 3:2, uniformly mixing to obtain an undried wet non-spore microbial agent;
(3) And (3) microbial inoculum drying: and (3) putting the non-spore microbial agent obtained in the step (2) into a baking oven with the temperature of 30 ℃ and drying for 18 hours to obtain the powdery non-spore microbial agent with the initial effective viable count of 85 hundred million/g.
Example 3
(1) Culturing bacterial liquid: culturing the strain liquid by adopting the alhagi pantoea XK-11, wherein the culture conditions are as follows: taking LB liquid culture, sterilizing at 121 ℃ for 20min in a high-temperature sterilizing pot, taking out the strain stored at-80 ℃ in advance to be completely dissolved, inoculating the strain into a sterile LB liquid culture medium, and culturing for 24h in a shaking table at 37 ℃ to obtain an activated strain for later use. Calculating the number of viable bacteria by adopting a dilution coating method to obtain non-spore microbial liquid with the initial effective viable bacteria number of 40 hundred million/ml;
(2) Bacterial liquid adsorption: adsorbing the non-spore microorganism bacterial liquid obtained in the step (1) by turfy soil according to the following steps of 4:3, uniformly mixing the materials in proportion to obtain an undried wet non-spore microbial agent preliminarily;
(3) And (3) microbial inoculum drying: and (3) putting the non-spore microbial agent obtained in the step (2) into a baking oven with the temperature of 40 ℃ for drying for 24 hours, and obtaining the powdery non-spore microbial agent with the initial effective viable count of 77 hundred million/g.
Example 4
(1) Culturing bacterial liquid: pseudomonas stutzeri NRCB010 is adopted for bacterial liquid culture, and the culture conditions are as follows: taking LB liquid culture, sterilizing at 121 ℃ for 20min in a high-temperature sterilizing pot, taking out the strain stored at-80 ℃ in advance to be completely dissolved, inoculating the strain into a sterile LB liquid culture medium, and culturing for 12h in a shaking table at 37 ℃ to obtain an activated strain for standby. Calculating the number of viable bacteria by adopting a dilution coating method to obtain non-spore microbial liquid with the initial effective viable bacteria number of 20 hundred million/ml;
(2) Bacterial liquid adsorption: adsorbing the non-spore microorganism bacterial liquid obtained in the step (1) by turfy soil according to the following steps of 2:1, uniformly mixing to obtain an undried wet non-spore microbial agent;
(3) And (3) microbial inoculum drying: and (3) putting the non-spore microbial agent obtained in the step (2) into a baking oven with the temperature of 20 ℃ for drying for 12 hours, and obtaining the powdery non-spore microbial agent with the initial effective viable count of 80 hundred million/g.
Example 5
(1) Culturing bacterial liquid: pseudomonas stutzeri NRCB010 is adopted for bacterial liquid culture, and the culture conditions are as follows: taking LB liquid culture, sterilizing at 121 ℃ for 20min in a high-temperature sterilizing pot, taking out the strain stored at-80 ℃ in advance to be completely dissolved, inoculating the strain into a sterile LB liquid culture medium, and culturing in a shaking table at 37 ℃ for 18h to obtain an activated strain for later use. Calculating the number of viable bacteria by adopting a dilution coating method to obtain non-spore microbial liquid with the initial effective viable bacteria number of 30 hundred million/ml;
(2) Bacterial liquid adsorption: adsorbing the non-spore microorganism bacterial liquid obtained in the step (1) by turfy soil according to the following steps of 3:2, uniformly mixing to obtain an undried wet non-spore microbial agent;
(3) And (3) microbial inoculum drying: and (3) putting the non-spore microbial agent obtained in the step (2) into a baking oven with the temperature of 30 ℃ and drying for 18 hours to obtain the powdery non-spore microbial agent with the initial effective viable count of 87 hundred million/g.
Example 6
(1) Culturing bacterial liquid: pseudomonas stutzeri NRCB010 is adopted for bacterial liquid culture, and the culture conditions are as follows: taking LB liquid culture, sterilizing at 121 ℃ for 20min in a high-temperature sterilizing pot, taking out the strain stored at-80 ℃ in advance to be completely dissolved, inoculating the strain into a sterile LB liquid culture medium, and culturing for 24h in a shaking table at 37 ℃ to obtain an activated strain for later use. Calculating the number of viable bacteria by adopting a dilution coating method to obtain non-spore microbial liquid with the initial effective viable bacteria number of 40 hundred million/ml;
(2) Bacterial liquid adsorption: adsorbing the non-spore microorganism bacterial liquid obtained in the step (1) by turfy soil according to the following steps of 4:3, uniformly mixing the materials in proportion to obtain an undried wet non-spore microbial agent preliminarily;
(3) And (3) microbial inoculum drying: and (3) putting the non-spore microbial agent obtained in the step (2) into a baking oven with the temperature of 40 ℃ for drying for 24 hours to obtain the powdery non-spore microbial agent with the initial effective viable count of 100 hundred million/g.
In order to verify that the turfy soil is a better carrier material for preparing the microbial inoculum, two groups of comparative tests for preparing the microbial inoculum respectively by adding the wheat bran carrier material are added.
Comparative example 1
(1) Culturing bacterial liquid: culturing the strain liquid by adopting the alhagi pantoea XK-11, wherein the culture conditions are as follows: taking LB liquid culture, sterilizing at 121 ℃ for 20min in a high-temperature sterilizing pot, taking out the strain stored at-80 ℃ in advance to be completely dissolved, inoculating the strain into a sterile LB liquid culture medium, and culturing for 24h in a shaking table at 37 ℃ to obtain an activated strain for later use. Calculating the number of viable bacteria by adopting a dilution coating method until the initial effective viable bacteria number is 40 hundred million/ml of non-spore microbial bacteria liquid;
(2) Bacterial liquid adsorption: adsorbing the non-spore microorganism bacterial liquid obtained in the step (1) by wheat bran according to the following steps of 4:3, uniformly mixing the materials in proportion to obtain an undried wet non-spore microbial agent preliminarily;
(3) And (3) microbial inoculum drying: and (3) putting the non-spore microbial agent obtained in the step (2) into a baking oven with the temperature of 40 ℃ for drying for 24 hours, and obtaining the powdery non-spore microbial agent with the initial effective viable count of 65 hundred million/g.
Comparative example 2
(1) Culturing bacterial liquid: pseudomonas stutzeri NRCB010 is adopted for bacterial liquid culture, and the culture conditions are as follows: taking LB liquid culture, sterilizing at 121 ℃ for 20min in a high-temperature sterilizing pot, taking out the strain stored at-80 ℃ in advance to be completely dissolved, inoculating the strain into a sterile LB liquid culture medium, and culturing in a shaking table at 37 ℃ for 18h to obtain an activated strain for later use. Calculating the number of viable bacteria by adopting a dilution coating method to obtain non-spore microbial liquid with the initial effective viable bacteria number of 30 hundred million/ml;
(2) Bacterial liquid adsorption: adsorbing the non-spore microorganism bacterial liquid obtained in the step (1) by wheat bran according to the following steps of 3:2, uniformly mixing to obtain an undried wet non-spore microbial agent;
(3) And (3) microbial inoculum drying: and (3) putting the non-spore microbial agent obtained in the step (2) into a baking oven with the temperature of 30 ℃ and drying for 18 hours to obtain the powdery non-spore microbial agent with the initial effective viable count of 70 hundred million/g.
Example 7 test for detection
(1) FIG. 1 shows the appearance of the preparation of Pantoea alhagi XK-11 microbial inoculum prepared in example 3 using peatmoss as the adsorption matrix, and the appearance of the other microbial inoculum (Pseudomonas stutzeri NRCB010 microbial inoculum) was the same.
(2) As shown in FIG. 2, the surface topography of the turfy soil before and after the loading of the strain is characterized by using a scanning electron microscope, the surface topography of the turfy soil before the first loading of the strain can be clearly observed to be in a scale microporous shape, which provides favorable conditions for the loading of the strain, and the second surface topography of the turfy soil after the turfy soil is loaded with the Alhagia camelina curcas XK-11 in example 3 can be clearly observed to be in a biofilm state development.
(3) FIG. 3 is a SEM surface morphology of the soil-loaded Pseudomonas stutzeri NRCB010 of example 5.
(4) As shown in fig. 4, SEM-EDS is used to characterize the elemental composition of the peatmoss, which corresponds to the elemental mass fraction composition in the table, and the results indicate that the peatmoss contains three elements in the content relationship: c (50.29%) > O (49.23%) > K (0.49%), and turfy soil can provide effective adsorption sites for the strain and also provide certain nutrient elements, thereby being beneficial to the adsorption survival of the strain.
(5) The powdery non-spore microbial agents prepared in examples 1 to 6 and comparative examples 1 to 2 were placed at room temperature of 25℃and periodically sampled, diluted and coated to calculate the number of effective colonies. The time required by the survival rate of the living bacteria in the microbial inoculum to be 10% is taken as a quality guarantee period index.
The two powder-type non-spore bacteria agents corresponding to examples 3 and 5 have the best normal temperature preservation effect, wherein the blank group of example 3 is a blank group of a common alhagi fungus XK-11 bacterial liquid and a sterile LB liquid culture medium according to the following formula 4:3, uniformly mixing the materials in proportion; the blank of example 5 is Pseudomonas stutzeri NRCB010 broth and sterile LB broth according to 3:2, and uniformly mixing the components.
As shown in FIG. 5 (A), the quality guarantee period of the powder type Pantoea alhagi XK-11 microbial inoculum of the experimental group is 180d, and the viable count is 9.6 hundred million/g; as shown in FIG. 5 (B), the blank group shelf life of the Pantoea alhagi XK-11 microbial inoculum is 5d, and the viable count is 2.7 hundred million/ml; compared with a blank group, the shelf life of the experimental group is effectively prolonged by 35 times. As shown in FIG. 6, the shelf life of a blank group of Pseudomonas stutzeri NRCB010 bacteria agent is 30d, and the viable count is 2.8 hundred million/ml; the shelf life of the powder type Pseudomonas stutzeri NRCB010 microbial inoculum of the experimental group is 360d, the viable count is 8.4 hundred million/g, and the quality is effectively improved by 11 times.
The wheat bran is used as a carrier material to prepare a microbial inoculum, the shelf life of the universal alhagi fungus XK-11 microbial inoculum prepared in comparative example 1 is 90d, and the viable count is 17 hundred million/g; the shelf life of the Pseudomonas stutzeri NRCB010 microbial inoculum prepared in comparative example 2 is 180d, and the viable count is 19 hundred million/g. The shelf life of the microbial inoculum is compared with that of the microbial inoculum corresponding to the comparative examples 1 and 2 and the shelf life of the microbial inoculum corresponding to the comparative examples 3 and 5 respectively, and the effective shelf life of the microbial inoculum is improved by 1 time in the examples 3 and 5. Compared with wheat bran, the turfy soil is an excellent carrier material capable of effectively prolonging the normal-temperature shelf life of the alhagi sparsifolia XK-11 and the Pseudomonas stutzeri NRCB010, can meet the shelf life requirement, and has practical application value.
The experiment shows that compared with two microbial agents (comparative examples 1 and 2) prepared from wheat bran at the normal temperature of 25 ℃, the shelf life effect of the two microbial agents prepared from turfy soil is best, wherein the shelf life effect of the alhage sparsifolia XK-11 microbial agent prepared in example 3 is best 180d, the shelf life is effectively improved by 35 times compared with a blank group, and the shelf life of the pseudomonas stutzeri NRCB010 microbial agent prepared in example 5 is effectively improved by 360d and 11 times compared with the blank group.
Example 8
The Alhagi sparsifolia XK-11 microbial inoculum (example 3) and the Pseudomonas stutzeri NRCB010 microbial inoculum (example 5) prepared from the turfy soil are respectively applied to pot experiments of rice, and the growth promotion effect of the microbial inoculum on crops is studied. The surfaces of rice seeds and tomato seeds were sterilized, soaked in 2.5% sodium hypochlorite solution for 5min, washed with sterile water several times, placed on a petri dish covered with sterile moist filter paper, germinated in a dark incubator at 30 ℃ for 2 days, and the germinated seeds were sown in flowerpots filled with 350g of field soil. Two bacterial suspensions (Alhagi, pantoea alhagi XK-11, pseudomonas stutzeri NRCB 010) were prepared before the experiment, and the concentration thereof was adjusted to 10 8 CFU/mL was ready for use. Selecting single colony of the Pantoea alhagi XK-11, placing the single colony in a fresh LB liquid medium, and culturing at 37 ℃ and 200rpm for 24 hours to obtain a suspension of the Pantoea alhagi XK-11. Selecting Pseudomonas stutzeri NRCB010 single colony to fresh LB liquid medium, placing in 37 ℃ and culturing at 200rpm for 18h to obtain Pseudomonas stutzeri NRCB010 suspension. The experimental design of the two experimental groups and the blank CK group is shown in table 1, and each treatment group was repeated three times. All flowerpots of the potting experiment are placed in a greenhouse at a temperature of 25 ℃ and an illumination time of 16:8h. After the plants grow for 5 weeks, harvesting, measuring the growth parameters, measuring the plant height and root length by using a metric ruler, then adjusting the temperature of the oven to 105 ℃, placing the plants into the oven for de-enzyming for 30 minutes, drying the overground part and the underground part of the plants in the oven at 60 ℃ until the weight is constant, weighing and recording data.
As is clear from Table 2, by comparing the growth parameters of rice seedlings in the CK group and the treatment group of example 3, it was found that the plant height, root length, aerial dry weight and underground dry weight of rice seedlings were significantly improved by the treatment with the Pantoea alhagi XK-11 microbial inoculum, the plant height of example 3 was significantly increased by 65.62%, the root length was significantly increased by 75.18%, the aerial dry weight was significantly increased by 200% and the underground dry weight was significantly increased by 164% as compared with CK. The result shows that the Pantoea alhagi XK-11 microbial inoculum can better promote the growth of rice.
As can be seen from table 3, by comparing the growth parameters of tomato seedlings in CK group and example 5 treatment group, it was found that the plant height, root length, dry weight on the ground and dry weight under the ground of tomato seedlings were significantly increased by treatment with pseudomonas stutzeri NRCB010 bacterial agent, and the plant height in example 5 was significantly increased by 27.40%, root length was significantly increased by 85.24%, dry weight on the ground was significantly increased by 300%, and dry weight under the ground was significantly increased by 171% as compared with CK. The pseudomonas stutzeri NRCB010 microbial inoculum is shown to be capable of better promoting tomato growth.
In conclusion, three microbial agents with the best normal-temperature shelf life effect, which are prepared from turfy soil, are the alhagi sparsifolia XK-11 microbial agents (example 3) and the Pseudomonas stutzeri NRCB010 microbial agents (example 5), and the results show that the two microbial agents can effectively promote the growth of rice and tomatoes respectively when applied to corresponding crop potting experiments, and have wide practical application values.
Table 1 potted plant experiment settings for rice and tomato
TABLE 2 Effect of Pantoea Alhagi XK-11 inoculant on rice seedling growth
TABLE 3 influence of Pseudomonas stutzeri NRCB010 inoculant on tomato seedling growth
The invention provides a non-spore microbial agent for prolonging the shelf life and a preparation method thereof, and the method and the way for realizing the technical scheme are numerous, the above is only a preferred embodiment of the invention, and it should be pointed out that a plurality of improvements and modifications can be made to a person skilled in the art without departing from the principle of the invention, and the improvements and modifications are also considered as the protection scope of the invention. The components not explicitly described in this embodiment can be implemented by using the prior art.
Claims (8)
1. The application of the turfy soil in prolonging the shelf life of the non-spore microbial agent is characterized in that the non-spore microbial agent comprises non-spore microbial liquid and turfy soil;
the non-spore microbial bacterial liquid is bacterial liquid of any one or more bacteria in the Pantoea alhagi and Pseudomonas stutzeri;
the preparation method of the non-spore microbial agent comprises the following steps:
s1: carrying out aerobic culture on any one or more bacteria in the alhagi pantoea and the pseudomonas stutzeri respectively to obtain non-spore microbial bacteria liquid;
s2: uniformly stirring the non-spore microbial liquid obtained in the step S1 with turfy soil to obtain an undried wet non-spore microbial agent;
s3: drying the undried wet non-spore microbial agent obtained in the step S2 to obtain the non-spore microbial agent;
in the step S3, the drying is carried out in an oven with the temperature of 20-40 ℃ for 12-24 hours.
2. The use according to claim 1, wherein the mass contents of the non-spore microbial liquid and the turfy soil in the non-spore microbial agent are 20% -50% and 50% -80%, respectively.
3. The use according to claim 1, wherein the viable count of the non-spore microbial agent is 70 to 100 hundred million/g.
4. The use according to claim 1, wherein when the non-spore microbial liquid is a liquid of pantoea alhagi, the viable count of the non-spore microbial liquid is 9.6 v/g or more after 180 days.
5. The use according to claim 1, wherein when the non-spore microbial liquid is a liquid of pseudomonas stutzeri, the viable count of the non-spore microbial agent is 8.4 v/g or more after 180 days.
6. The use according to claim 1, wherein in step S1, the mass ratio of the non-spore microorganism bacterial liquid to the turfy soil is (0.5-3): 1.
7. The use according to claim 1, wherein in step S1, the mass ratio of the non-spore microbial liquid to the turfy soil is 2:1, 3:2, 4:3.
8. The method according to claim 5, wherein in the step S1, the viable count of the non-spore microorganism bacterium liquid is 20-40 hundred million/mL.
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| CN103408198A (en) * | 2013-08-23 | 2013-11-27 | 尹梦珍 | Sewage treatment method in landscape planning |
| WO2019218412A1 (en) * | 2018-05-15 | 2019-11-21 | 中国环境科学研究院 | Immobilized microbial agent for in situ remediation of contaminated sediments, preparation method and use |
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| CN103408198A (en) * | 2013-08-23 | 2013-11-27 | 尹梦珍 | Sewage treatment method in landscape planning |
| WO2019218412A1 (en) * | 2018-05-15 | 2019-11-21 | 中国环境科学研究院 | Immobilized microbial agent for in situ remediation of contaminated sediments, preparation method and use |
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