CN115029288A - Bacillus subtilis composite hydraulic microbial agent for water and soil conservation and preparation method thereof - Google Patents
Bacillus subtilis composite hydraulic microbial agent for water and soil conservation and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a bacillus subtilis composite hydraulic microbial inoculum for water and soil conservation and a preparation method thereof, wherein the bacillus subtilis composite hydraulic microbial inoculum comprises the following components in parts by weight: 30-80 parts of bacillus subtilis live spore powder, 15-40 parts of bacillus thuringiensis live spore powder, 15-40 parts of lactobacillus casei live spore powder, 15-30 parts of cereal husk hydrolysate and 40-100 parts of long-acting slow-release bacteria storage gel composition. The invention belongs to the technical field of compound hydraulic fungicide; according to the invention, by adding the long-acting slow-release bacteria storage gel composition, the propagation condition of the bacillus subtilis in a complex environment is improved, and the survival activity and the stress resistance activity of the bacillus subtilis and auxiliary strains thereof are improved; meanwhile, the grain husk hydrolysate forms a component which can be directly absorbed by plants through the degradation of bacillus subtilis and the degradation of auxiliary strains, so that the growth of the plants is improved.
Description
Technical Field
The invention belongs to the technical field of composite hydraulic microbial inoculum, and particularly relates to a bacillus subtilis composite hydraulic microbial inoculum for water and soil conservation and a preparation method thereof.
Background
Water and soil conservation refers to the prevention, protection, improvement and reasonable utilization of water and soil resources in mountainous areas, hilly areas and sand storm areas, maintenance and improvement of land productivity, so as to bring the economic and social benefits of water and soil resources into full play and establish comprehensive scientific technology with good ecological environment The safety of cities and towns is significant.
The damage of vegetation is the important reason of soil erosion, and the erosion influence of earth's surface vegetation cover to ground especially domatic soil is very big, and the vegetation has the slowing down effect to the runoff velocity of flow, thereby the change of soil structure can increase infiltration and keep in good care the water source in addition, improves the anti erosion effect of soil.
Bacillus subtilis is a kind of Bacillus, and can decompose tryptophan to form indole by using protein, various sugars and starch, the bacillus subtilis is widely distributed in soil and putrefactive organic matters, is also a commonly used biological control and soil improvement strain, but the application of the bacillus subtilis in vegetation maintenance and water and soil conservation protection is not seen at present, in addition, the problems of the prior art are mainly that due to the complexity of the external physical environment, the bacillus subtilis applied in the soil can not realize long-acting effect, the water and soil loss treatment engineering quantity is huge, and if the bacillus subtilis is applied for a plurality of times, the workload is increased, so the bacillus subtilis can not be popularized, secondly, the existing water and soil loss coagulation microbial inoculum cannot realize the purposes of adjusting the soil structure and improving the plant growth, and further cannot radically form the ecological stability which is easy to cause water and soil loss.
Disclosure of Invention
Aiming at the situation and overcoming the defects of the prior art, the invention provides the bacillus subtilis composite hydraulic microbial inoculum for water and soil conservation and the preparation method thereof, aiming at solving the problem that the composite hydraulic microbial inoculum cannot realize long-acting effect, the invention improves the propagation condition of the bacillus subtilis in a complex environment by adding the long-acting slow-release bacteria storage gel composition, and improves the survival activity and the anti-stress activity of the bacillus subtilis and auxiliary strains thereof by using the traditional Chinese medicine powder composition, thereby improving the use applicability of the composite hydraulic microbial inoculum; meanwhile, the grain husk hydrolysate is rich in various substances required by plant growth, and components capable of being directly absorbed by plants are formed through the degradation of bacillus subtilis and the degradation of auxiliary strains, so that the growth of the plants is improved, and the prevention and control effects on water and soil conservation are further enhanced.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows: the invention provides a bacillus subtilis composite hydraulic microbial inoculum for water and soil conservation, which comprises the following components in parts by weight:
30-80 parts of a bacillus subtilis live spore powder agent;
15-40 parts of a bacillus thuringiensis live spore powder;
15-40 parts of lactobacillus casei live spore powder;
15-30 parts of grain husk hydrolysate;
40-100 parts of long-acting slow-release bacteria storage gel composition.
Further, the compound hydraulic bacterial agent comprises the following components in parts by weight:
40-60 parts of bacillus subtilis active spore powder;
20-30 parts of bacillus thuringiensis live spore powder;
20-30 parts of lactobacillus casei live spore powder;
15-20 parts of grain husk hydrolysate;
50-80 parts of long-acting slow-release bacterium storage gel composition.
Preferably, the long-acting slow-release bacteria storage gel composition comprises the following components in parts by weight:
10-30 parts of curdlan;
1-15 parts of a Chinese medicinal powder composition;
5-20 parts of trehalose;
1-15 parts of beta-cyclodextrin.
In order to better achieve the purpose of the invention, as a further preferable mode, the long-acting slow-release bacteria storage gel composition comprises the following components in parts by weight:
15-20 parts of curdlan;
5-10 parts of a traditional Chinese medicine powder composition;
10-15 parts of trehalose;
5-10 parts of beta-cyclodextrin.
Further, the grain husk hydrolysate is one or more of peanut husk hydrolysate, rice husk hydrolysate, soybean husk hydrolysate, and wheat husk hydrolysate.
Further, the traditional Chinese medicine powder composition is prepared from astragalus powder, purslane powder, rhizoma smilacis glabrae powder and betel nut powder according to a proportion of 1: 1: 0.8: 1, mixing and preparing.
In addition, the invention also provides a preparation method of the bacillus subtilis composite hydraulic microbial agent for water and soil conservation, which comprises the following steps:
(1) preparing bacillus subtilis active spore powder, bacillus thuringiensis active spore powder and lactobacillus casei active spore powder;
(2) preparation of cereal husk hydrolysate: will be provided withPulverizing cereal husk with pulverizer, filtering with 60-100 mesh sieve to obtain cereal husk powder, adding 1-2% by volume of H 2 SO 4 Hydrolyzing in the solution at 121 deg.C for 120min, cooling, adjusting pH to 5-8, centrifuging, and collecting supernatant which is grain husk hydrolysate;
(3) preparing a long-acting slow-release bacterium storage gel composition: taking radix astragali powder, purslane powder, rhizoma smilacis glabrae powder and betel nut powder according to the proportion of 1: 1: 0.8: 1, mixing the materials with curdlan, trehalose and beta-cyclodextrin according to a conventional gel preparation process to prepare a long-acting slow-release bacteria storage gel composition;
(4) and fully mixing the bacillus subtilis active spore powder, the bacillus thuringiensis active spore powder and the lactobacillus casei active spore powder with the grain husk hydrolysate, and adding the mixture into the long-acting slow-release bacteria storage gel composition to obtain the composite hydraulic microbial agent.
Preferably, the preparation method of the bacillus subtilis active spore powder comprises the following steps:
(1) activating a bacterial liquid: diluting Bacillus subtilis by 5-10 times, coating on activated solid culture medium, and culturing at 37 deg.C for 12 hr to obtain activated Bacillus subtilis;
(2) strain amplification activation culture: selecting a single colony of the activated bacillus subtilis by using an inoculating needle, and culturing in 500mL of propagation liquid culture medium at 37 ℃ and 180rpm for 12h to obtain the bacillus subtilis subjected to propagation;
(3) enrichment culture of bacterial liquid: sucking 5mL of the bacillus subtilis subjected to the amplification culture in the second step into an amplification liquid culture medium, and culturing at 37 ℃ and 180rpm for 24-48h to obtain a bacillus subtilis culture solution;
(4) powder preparation: centrifuging the Bacillus subtilis culture solution at 150rpm for 1min, removing supernatant to obtain Bacillus subtilis thallus, and oven drying at 30-37 deg.C for 30-60min to obtain Bacillus subtilis active spore powder.
Preferably, the preparation method of the bacillus thuringiensis active spore powder comprises the following steps:
(1) activating a bacterial liquid: diluting Bacillus thuringiensis by 5-10 times, coating on an activated solid culture medium, and culturing at 37 deg.C for 12 hr to obtain activated Bacillus thuringiensis;
(2) and (3) strain amplification activation culture: selecting a single colony of the activated bacillus thuringiensis by using an inoculating needle, and culturing in 500mL of propagation liquid culture medium at 37 ℃ and 180rpm for 12h to obtain the bacillus thuringiensis through propagation;
(3) enrichment culture of bacterial liquid: sucking 5mL of the bacillus thuringiensis in the expanded propagation liquid culture medium in the second step, and culturing at 37 ℃ and 180rpm for 24-48h to obtain a bacillus thuringiensis culture solution;
(4) powder preparation: centrifuging the Bacillus thuringiensis culture solution at 150rpm for 1min, removing supernatant to obtain Bacillus thuringiensis thallus, and oven drying at 30-37 deg.C for 30-60min to obtain Bacillus thuringiensis active spore powder.
Preferably, the preparation method of the lactobacillus casei active spore powder comprises the following steps:
(1) activating a bacterial liquid: diluting lactobacillus casei by 5-10 times, coating on an activated solid culture medium, and culturing at 37 deg.C for 12 hr to obtain activated lactobacillus casei;
(2) and (3) strain amplification activation culture: selecting a single colony of activated lactobacillus casei by using an inoculating needle, and culturing in 500mL of propagation liquid culture medium at 37 ℃ and 180rpm for 12h to obtain the expanded cultured lactobacillus casei;
(3) enrichment culture of bacterial liquid: sucking 5mL of the lactobacillus casei subjected to the expanded culture in the step II into an expanded propagation liquid culture medium, and culturing at 37 ℃ and 180rpm for 24-48h to obtain a lactobacillus casei culture solution;
(4) powder preparation: centrifuging the lactobacillus casei culture solution at 150rpm for 1min, removing the supernatant to obtain lactobacillus casei thallus, and oven drying at 30-37 deg.C for 30-60min to obtain lactobacillus casei active spore powder.
The invention with the structure has the following beneficial effects:
(1) in order to solve the problem that the compound hydraulic fungicide cannot realize long-acting effect, the invention improves the propagation condition of the bacillus subtilis in a complex environment by adding the long-acting slow-release bacterium storage gel composition;
(2) the use of the traditional Chinese medicine powder composition improves the survival activity and the stress resistance activity of the bacillus subtilis and the auxiliary bacterial strains thereof, thereby improving the use applicability;
(3) the astragalus powder has the function of improving the resistance of the bacillus subtilis;
(4) in addition, the bacillus thuringiensis, the lactobacillus casei and the bacillus subtilis have a synergistic effect, and the effect of promoting plant growth is realized;
(5) meanwhile, the grain husk hydrolysate is rich in various substances required by plant growth, and components which can be directly absorbed by plants are formed through the degradation of bacillus subtilis and the degradation of auxiliary strains, so that the growth of the plants is improved, the control effect on water and soil conservation is further enhanced,
(6) the composite water flocculant provided by the invention can be applied to water and soil conservation in various environments, and has important values for reducing water and soil loss and forming a stable ecological environment.
Drawings
FIG. 1 is a graph showing the measurement results of a Bacillus subtilis composite hydraulic fungicide for soil and water conservation for maintaining the slope length of a slope;
FIG. 2 is a graph showing the results of measurements of a Bacillus subtilis complex hydraulic preparation for soil and water conservation used for maintaining the slope of a slope;
FIG. 3 is a diagram showing the measurement results of a Bacillus subtilis complex hydraulic inoculant for soil and water conservation for promoting the growth of plant roots;
FIG. 4 is a graph showing the results of measurements of the growth of bacterial colonies promoted by a Bacillus subtilis complex hydraulic inoculant for soil and water conservation.
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments; all other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.
The experimental procedures in the following examples are all conventional ones unless otherwise specified.
The test materials, culture media, and test strains used in the following examples were all purchased from commercial sources unless otherwise specified, wherein in the present examples, Bacillus subtilis was purchased from the labor-saving culture Collection in Sichuan (strain Collection No. SICC 1.1325), Bacillus thuringiensis was purchased from the labor-saving culture Collection in Sichuan (strain Collection No. SCTCC 100350), and Lactobacillus casei was purchased from the labor-saving culture Collection in Sichuan (strain Collection No. SICC 1.1271).
Example 1
Bacillus subtilis composite hydraulic microbial agent for water and soil conservation
The compound hydraulic fungicide comprises the following components in parts by weight:
50 parts of bacillus subtilis active spore powder;
20 parts of bacillus thuringiensis and live spore powder;
20 parts of lactobacillus casei live spore powder;
15 parts of grain husk hydrolysate;
55 parts of long-acting slow-release bacteria storage gel composition.
The long-acting slow-release bacterium storage gel composition comprises the following components in parts by weight:
20 parts of curdlan;
8 parts of a traditional Chinese medicine powder composition;
12 parts of trehalose;
6 parts of beta-cyclodextrin.
Wherein the grain husk hydrolysate is prepared from peanut husk hydrolysate, rice husk hydrolysate and soybean husk hydrolysate in a weight ratio of 1: 1: 2, preparing the components in a combined manner; the traditional Chinese medicine powder composition is prepared from astragalus powder, purslane powder, glabrous greenbrier rhizome powder and betel nut powder according to the proportion of 1: 1: 0.8: 1, mixing and preparing.
In addition, the invention also provides a preparation method of the bacillus subtilis composite hydraulic microbial agent for water and soil conservation, which comprises the following steps:
(1) preparing bacillus subtilis active spore powder, bacillus thuringiensis active spore powder and lactobacillus casei active spore powder;
(2) preparation of cereal husk hydrolysate: pulverizing cereal husk powder with pulverizer, filtering with 100 mesh sieve to obtain cereal husk powder, adding 2% H 2 SO 4 Hydrolyzing in the solution at 121 deg.C for 120min, cooling, adjusting pH to 5-6, centrifuging, and collecting supernatant which is grain husk hydrolysate;
(3) preparing a long-acting slow-release bacterium storage gel composition: taking radix astragali powder, purslane powder, rhizoma smilacis glabrae powder and betel nut powder according to the proportion of 1: 1: 0.8: 1, mixing the materials with curdlan, trehalose and beta-cyclodextrin according to a conventional gel preparation process to prepare a long-acting slow-release bacteria storage gel composition;
(4) and fully mixing the bacillus subtilis active spore powder, the bacillus thuringiensis active spore powder and the lactobacillus casei active spore powder with the grain husk hydrolysate, and adding the mixture into the long-acting slow-release bacteria storage gel composition to obtain the composite hydraulic microbial agent.
The preparation method of the bacillus subtilis active spore powder comprises the following steps:
(1) activating a bacterial liquid: diluting bacillus subtilis by 5 times, coating the diluted bacillus subtilis on an activated solid culture medium, and culturing at 37 ℃ for 12h to obtain activated bacillus subtilis;
(2) and (3) strain amplification activation culture: selecting a single colony of the activated bacillus subtilis by using an inoculating needle, and culturing in 500mL of propagation liquid culture medium at 37 ℃ and 180rpm for 12h to obtain the propagation-cultured bacillus subtilis;
(3) enrichment culture of bacterial liquid: sucking 5mL of the bacillus subtilis subjected to the amplification culture in the second step into an amplification propagation liquid culture medium, and culturing for 24-48h at 37 ℃ and 180rpm to obtain a bacillus subtilis culture solution;
(4) powder preparation: centrifuging the Bacillus subtilis culture solution at 150rpm for 1min, removing supernatant to obtain Bacillus subtilis thallus, and oven drying at 30-37 deg.C for 30-60min to obtain Bacillus subtilis powder.
The preparation method of the bacillus thuringiensis active spore powder comprises the following steps:
(1) activating a bacterial liquid: diluting Bacillus thuringiensis by 5 times, coating the diluted Bacillus thuringiensis on an activated solid culture medium, and culturing at 37 ℃ for 12h to obtain activated Bacillus thuringiensis;
(2) and (3) strain amplification activation culture: selecting a single colony of the activated bacillus thuringiensis by using an inoculating needle, and culturing in 500mL of propagation liquid culture medium at 37 ℃ and 180rpm for 12h to obtain the bacillus thuringiensis through propagation;
(3) enrichment culture of bacterial liquid: sucking 5mL of the bacillus thuringiensis in the expanded propagation liquid culture medium in the second step, and culturing at 37 ℃ and 180rpm for 24-48h to obtain a bacillus thuringiensis culture solution;
(4) powder preparation: centrifuging the Bacillus thuringiensis culture solution at 150rpm for 1min, removing supernatant to obtain Bacillus thuringiensis thallus, and oven drying at 30-37 deg.C for 30-60min to obtain Bacillus thuringiensis active spore powder.
The preparation method of the lactobacillus casei active spore powder comprises the following steps:
(1) activating bacterial liquid: diluting lactobacillus casei by 5 times, coating on an activated solid culture medium, and culturing at 37 ℃ for 12h to obtain activated lactobacillus casei;
(2) and (3) strain amplification activation culture: selecting a single colony of activated lactobacillus casei by using an inoculating needle, and culturing in 500mL of propagation liquid culture medium at 37 ℃ and 180rpm for 12h to obtain the expanded cultured lactobacillus casei;
(3) enrichment culture of bacterial liquid: sucking 5mL of the lactobacillus casei subjected to the expanding culture in the step II into an expanding propagation liquid culture medium, and culturing for 24-48h at 37 ℃ and 180rpm to obtain a lactobacillus casei culture solution;
(4) powder preparation: centrifuging the lactobacillus casei culture solution at 150rpm for 1min, removing supernatant to obtain lactobacillus casei thallus, and oven drying at 30-37 deg.C for 30-60min to obtain lactobacillus casei active spore powder.
Example 2
Bacillus subtilis composite hydraulic microbial agent for water and soil conservation
The compound hydraulic microbial inoculum comprises the following components in parts by weight:
55 parts of a bacillus subtilis live spore powder;
20 parts of bacillus thuringiensis live spore powder;
25 parts of lactobacillus casei active spore powder;
20 parts of grain husk hydrolysate;
60 parts of long-acting slow-release bacterium storage gel composition.
The long-acting slow-release bacterium storage gel composition comprises the following components in parts by weight:
15 parts of curdlan;
7 parts of a Chinese medicinal powder composition;
12 parts of trehalose;
8 parts of beta-cyclodextrin.
Wherein the grain husk hydrolysate is prepared from peanut husk hydrolysate, rice husk hydrolysate and soybean husk hydrolysate in a weight ratio of 1: 1: 2, preparing the components in a combined manner; the traditional Chinese medicine powder composition is prepared from astragalus powder, purslane powder, rhizoma smilacis glabrae powder and betel nut powder according to the proportion of 1: 1: 0.8: 1, mixing and preparing.
In addition, the invention also provides a preparation method of the bacillus subtilis composite hydraulic microbial inoculum for water and soil conservation, and the preparation method is implemented by referring to the example 1.
Example 3
Bacillus subtilis composite hydraulic microbial agent for water and soil conservation
The compound hydraulic microbial inoculum comprises the following components in parts by weight:
55 parts of a bacillus subtilis live spore powder;
15 parts of bacillus thuringiensis live spore powder;
25 parts of lactobacillus casei active spore powder;
15 parts of grain husk hydrolysate;
55 parts of long-acting slow-release bacterium storage gel composition.
The long-acting slow-release bacterium storage gel composition comprises the following components in parts by weight:
18 parts of curdlan;
10 parts of a traditional Chinese medicine powder composition;
12 parts of trehalose;
6 parts of beta-cyclodextrin.
Wherein the grain husk hydrolysate is prepared from peanut husk hydrolysate, rice husk hydrolysate and soybean husk hydrolysate in a weight ratio of 1: 1: 2, preparing the components in a combined manner; the traditional Chinese medicine powder composition is prepared from astragalus powder, purslane powder, glabrous greenbrier rhizome powder and betel nut powder according to the proportion of 1: 1: 0.8: 1, mixing and preparing.
In addition, the invention also provides a preparation method of the bacillus subtilis composite hydraulic microbial inoculum for water and soil conservation, and the preparation method is implemented by referring to the example 1.
Comparative example 1
The comparative example provides a bacillus subtilis composite hydraulic microbial inoculum for water and soil conservation and a preparation method thereof, and the bacillus subtilis composite hydraulic microbial inoculum is different from the bacillus subtilis composite hydraulic microbial inoculum in example 1 only in that all components do not contain a long-acting slow-release bacteria storage gel composition, and the rest components and the component content are the same as those in example 1.
Examples of the experiments
Evaluation of water and soil conservation effect and long-term effect of bacillus subtilis composite hydraulic fungicide for water and soil conservation
(1) Influence of composite hydraulic microbial inoculum on length and gradient of slope
The method comprises the steps of mixing the compound hydraulic fungicide prepared in the embodiment 1, the embodiment 2, the embodiment 3 and the comparative example 1 with basic forage seeds, spraying and throwing the mixture onto a slope, wherein the compound hydraulic fungicide is not added to serve as a blank control group, 3 test cells are arranged in each of the test group and the blank control group, the slope length and the slope gradient of each test cell are measured, the slope length and the slope gradient are measured again after the rainy season is finished, the slope gradient is the percentage of the ratio of the elevation difference to the horizontal distance, and the slope length change value and the slope gradient change value are calculated, wherein the slope length difference value is the difference value between the slope length after the test and the slope length before the test, and the slope gradient difference value is the difference value between the slope gradient before the test and the slope after the test.
As shown in fig. 1 and fig. 2, compared with the blank control group, the use of the compound hydraulic fungicide can reduce the influence of rainwater on the length and the gradient of the slope, thereby avoiding soil erosion on the slope.
(2) Effect of compound hydrogel inoculant on promoting plant growth
The root growth conditions of the plants in the test cells are sampled and measured, as shown in fig. 3, the root systems of the plants in the blank control groups are smaller than those in the test groups, and the fact that the composite hydraulic fungicide is used can improve the growth of the plants is shown, so that water and soil conservation is facilitated.
(3) Quantitative analysis of soil bacteria flora
Sampling the soil of the experimental community, and measuring the number of the soil bacterial flora in 0 month, 1 month, 2 months, 3 months, 4 months and 5 months by adopting a flat plate counting method, wherein the experimental steps of the flat plate counting method are as follows: collecting soil of a test site by adopting a five-point sampling method, filtering by using three layers of filter paper, collecting filtrate, absorbing the same amount of the filtrate, diluting by 100 times and 1000 times, coating the filtrate on a beef extract peptone culture medium (containing 50 mu g/mL nystatin solution), and observing the number of soil bacterial flora by using a counter and a microscope.
As shown in fig. 4, the number of the soil bacteria flora is significantly increased compared to the initial time (0 month), the number of the soil bacteria flora can be increased by using the composite hydraulic microbial inoculum compared to the blank control, and examples 1, 2 and 3 can still have a very high number of the soil bacteria flora at 5 months compared to the comparative example, which indicates that the long-acting slow-release bacterial storage gel composition can increase the action time of the composite hydraulic microbial inoculum.
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 alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
The invention and its embodiments have been described above, without limitation, and what is shown in the drawings is only one of the embodiments of the invention, and the practical application is not limited thereto. In summary, those skilled in the art should appreciate that they can readily use the disclosed conception and specific embodiments as a basis for designing or modifying other structures for carrying out the same purposes of the present invention without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (10)
1. A bacillus subtilis composite hydraulic microbial inoculum for water and soil conservation is characterized in that: the compound hydraulic microbial inoculum comprises the following components in parts by weight:
30-80 parts of a bacillus subtilis live spore powder agent;
15-40 parts of bacillus thuringiensis live spore powder;
15-40 parts of lactobacillus casei live spore powder;
15-30 parts of grain husk hydrolysate;
40-100 parts of long-acting slow-release bacteria storage gel composition.
2. The bacillus subtilis complex hydraulic inoculant for water and soil conservation as claimed in claim 1, wherein: the compound hydraulic microbial inoculum comprises the following components in parts by weight:
40-60 parts of bacillus subtilis active spore powder;
20-30 parts of bacillus thuringiensis live spore powder;
20-30 parts of lactobacillus casei live spore powder;
15-20 parts of grain husk hydrolysate;
50-80 parts of long-acting slow-release bacterium storage gel composition.
3. The bacillus subtilis complex hydraulic inoculant for water and soil conservation as claimed in claim 2, wherein: the long-acting slow-release bacterium storage gel composition comprises the following components in parts by weight:
10-30 parts of curdlan;
1-15 parts of a Chinese medicinal powder composition;
5-20 parts of trehalose;
1-15 parts of beta-cyclodextrin.
4. The bacillus subtilis complex hydraulic inoculant for water and soil conservation as claimed in claim 3, wherein: the long-acting slow-release bacterium storage gel composition comprises the following components in parts by weight:
15-20 parts of curdlan;
5-10 parts of a traditional Chinese medicine powder composition;
10-15 parts of trehalose;
5-10 parts of beta-cyclodextrin.
5. The Bacillus subtilis complex hydraulic inoculant according to claim 4, wherein: the grain husk hydrolysate is one or more of peanut husk hydrolysate, rice husk hydrolysate, soybean husk hydrolysate, and wheat husk hydrolysate.
6. The Bacillus subtilis complex hydraulic microbial agent for soil and water conservation as claimed in claim 5, wherein: the traditional Chinese medicine powder composition comprises astragalus powder, purslane powder, rhizoma smilacis glabrae powder and betel nut powder according to the proportion of 1: 1: 0.8: 1, mixing and preparing.
7. The method for preparing the bacillus subtilis complex hydraulic inoculant for water and soil conservation as claimed in claim 6, wherein the bacillus subtilis complex hydraulic inoculant comprises the following steps: the preparation method comprises the following steps:
(1) preparing bacillus subtilis active spore powder, bacillus thuringiensis active spore powder and lactobacillus casei active spore powder;
(2) preparation of cereal husk hydrolysate: pulverizing cereal husk powder with pulverizer, filtering with 60-100 mesh sieve to obtain cereal husk powder, adding 1-2% by volume of H 2 SO 4 Hydrolyzing in the solution at 121 deg.C for 120min, cooling, adjusting pH to 5-8, centrifuging, and collecting supernatant which is grain husk hydrolysate;
(3) preparing a long-acting slow-release bacterium storage gel composition: taking radix astragali powder, purslane powder, rhizoma smilacis glabrae powder and betel nut powder according to the proportion of 1: 1: 0.8: 1 mixing to obtain a traditional Chinese medicine powder composition, and then mixing with curdlan, trehalose and beta-cyclodextrin according to a conventional gel preparation process to prepare a long-acting slow-release bacteria storage gel composition;
(4) and mixing the bacillus subtilis active-spore powder, the bacillus thuringiensis active-spore powder and the lactobacillus casei active-spore powder with the cereal husk hydrolysate, and adding the mixture into the long-acting slow-release bacterium storage gel composition to obtain the composite hydrogel agent.
8. The method for preparing a bacillus subtilis complex hydraulic inoculant for water and soil conservation as claimed in claim 7, wherein the bacillus subtilis complex hydraulic inoculant comprises: the preparation method of the bacillus subtilis active spore powder comprises the following steps:
(1) activating bacterial liquid: diluting Bacillus subtilis by 5-10 times, coating on activated solid culture medium, and culturing at 37 deg.C for 12 hr to obtain activated Bacillus subtilis;
(2) and (3) strain amplification activation culture: selecting a single colony of the activated bacillus subtilis by using an inoculating needle, and culturing in 500mL of propagation liquid culture medium at 37 ℃ and 180rpm for 12h to obtain the bacillus subtilis subjected to propagation;
(3) enrichment culture of bacterial liquid: sucking 5mL of the bacillus subtilis for the enlarged culture in an enlarged propagation liquid culture medium, and culturing at 37 ℃ and 180rpm for 24-48h to obtain a bacillus subtilis culture solution;
(4) powder preparation: centrifuging the Bacillus subtilis culture solution at 150rpm for 1min, removing supernatant to obtain Bacillus subtilis thallus, and oven drying at 30-37 deg.C for 30-60min to obtain Bacillus subtilis powder.
9. The method for preparing a bacillus subtilis complex hydraulic inoculant for water and soil conservation as claimed in claim 8, wherein the bacillus subtilis complex hydraulic inoculant comprises: the preparation method of the bacillus thuringiensis active spore powder comprises the following steps:
(1) activating a bacterial liquid: diluting Bacillus thuringiensis by 5-10 times, coating on an activated solid culture medium, and culturing at 37 deg.C for 12 hr to obtain activated Bacillus thuringiensis;
(2) and (3) strain amplification activation culture: selecting a single colony of the activated bacillus thuringiensis by using an inoculating needle, and culturing in 500mL of propagation liquid culture medium at 37 ℃ and 180rpm for 12h to obtain the bacillus thuringiensis through propagation;
(3) enrichment culture of bacterial liquid: sucking 5mL of the bacillus thuringiensis subjected to the amplification culture into an amplification propagation liquid culture medium, and culturing at 37 ℃ and 180rpm for 24-48h to obtain a bacillus thuringiensis culture solution;
(4) powder preparation: centrifuging the Bacillus thuringiensis culture solution at 150rpm for 1min, removing supernatant to obtain Bacillus thuringiensis thallus, and oven drying at 30-37 deg.C for 30-60min to obtain Bacillus thuringiensis active spore powder.
10. The method for preparing a bacillus subtilis complex hydraulic inoculant for water and soil conservation as claimed in claim 9, wherein the bacillus subtilis complex hydraulic inoculant comprises: the preparation method of the lactobacillus casei active spore powder comprises the following steps:
(1) activating a bacterial liquid: diluting lactobacillus casei by 5-10 times, coating on an activated solid culture medium, and culturing at 37 deg.C for 12 hr to obtain activated lactobacillus casei;
(2) and (3) strain amplification activation culture: selecting a single colony of activated lactobacillus casei by using an inoculating needle, and culturing in 500mL of propagation liquid culture medium at 37 ℃ and 180rpm for 12h to obtain the expanded cultured lactobacillus casei;
(3) enrichment culture of bacterial liquid: sucking 5mL of the lactobacillus casei subjected to the expanded culture into an expanded propagation liquid culture medium, and culturing at 37 ℃ and 180rpm for 24-48h to obtain a lactobacillus casei culture solution;
(4) powder preparation: centrifuging the lactobacillus casei culture solution at 150rpm for 1min, removing supernatant to obtain lactobacillus casei thallus, and oven drying at 30-37 deg.C for 30-60min to obtain lactobacillus casei active spore powder.
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