CN108715821B - Microbial compound inoculant and preparation method and application thereof - Google Patents
Microbial compound inoculant and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a microbial compound inoculant which is prepared from the following raw materials in parts by weight: 30-50 parts of bacillus subtilis fermented bacterial liquid, 20-30 parts of bacillus mycoides fermented bacterial liquid, 10-20 parts of corallina nocardia fermented bacterial liquid, 10-20 parts of functional microbial inoculum and 60-80 parts of carrier. In addition, the invention also provides a preparation method and application of the microbial compound inoculant. The microbial compound microbial inoculum prepared by the invention contains enough effective viable bacteria, can stably and efficiently play a role, can obviously increase the content of soil organic matters and nutrient elements such as nitrogen, phosphorus, potassium and the like, improves the nutrient conversion rate and the number of beneficial microorganisms in the product, promotes soil remediation, increases the soil fertility, can reduce the using amount of chemical fertilizers, simultaneously enhances the disease resistance of crops, improves the quality of the crops, reduces the investment, improves the yield and promotes the income increase.
Description
Technical Field
The invention belongs to the technical field of agricultural microorganisms, and particularly relates to a microbial compound inoculant as well as a preparation method and application thereof.
Background
Agricultural fertilizers include chemical fertilizers and microbial fertilizers, and microbial fertilizers are products which are used for obtaining specific fertilizer effects under the action of living activities of microorganisms, and are different from traditional fertilizers and organic fertilizers in nature. The microbial fertilizer is prepared by mixing the microbial agent and the microbial fertilizer formula, has low production cost, convenient use and good application effect, improves the soil environment, does not pollute the environment, increases the yield after being applied, and can improve the quality of agricultural products and reduce the application amount of the fertilizer.
After the problems of soil hardening, fertility reduction, soil salinization, ecological destruction, environmental pollution and agricultural and sideline product quality reduction brought by chemical fertilizers are more and more prominent, the microbial fertilizer is rapidly developed. However, due to the reasons of strain and component proportion, the quality of products in the current market is uneven, some problems occur in production and application, the number of effective bacteria is not up to the standard, and the soil performance improvement and agricultural product yield increase effects in practical application are not ideal. The effective viable count in the microbial fertilizer is an important index of product quality, the effective viable count does not reach the standard, the product quality cannot be guaranteed, and the reflected application effect is unstable. However, the reason for the strains is not the only reason for affecting the effect of the fertilizer, and in fact, the mutual matching between the microbial strains and other components in the fertilizer also affects the effect of the fertilizer. Because the components have a mutual cooperation and antagonism relationship, the proper component proportion has a close relationship to the propagation and activity of microorganisms in the microbial fertilizer and the growth of crops. If the problem of strains can be solved, the relationship among the mixture ratios of the components is coordinated through innovation of the mixture ratios of the components of the fertilizer, so that a better effect is obtained, the yield is effectively improved, and a great innovation effect is played on the development of the field of microbial fertilizers.
Disclosure of Invention
The invention provides a microbial compound inoculant, and solves the problems that the effective viable count in microbial fertilizers in the prior art often does not reach the standard, the product quality is not guaranteed, and the application effect of products is unstable.
The invention aims to provide a microbial compound inoculant which is prepared from the following raw materials in parts by weight: 30-50 parts of bacillus subtilis fermented bacterial liquid, 20-30 parts of bacillus mycoides fermented bacterial liquid, 10-20 parts of corallina nocardia fermented bacterial liquid, 10-20 parts of functional microbial inoculum and 60-80 parts of carrier.
Preferably, the preparation raw materials of the microbial compound inoculant comprise the following components in parts by weight: 40 parts of bacillus subtilis zymocyte liquid, 25 parts of mushroom-shaped bacillus zymocyte liquid, 15 parts of coralline nocardia zymocyte liquid, 15 parts of functional microbial inoculum and 70 parts of carrier.
Preferably, the functional microbial inoculum is prepared from a azotobacter microbial inoculum and a rhizobium microbial inoculum according to the ratio of 3: 1, and the number of effective viable bacteria in the azotobacter and the rhizobium inoculant is more than or equal to 1010cfu/g；
Preferably, the number of effective viable bacteria in the bacillus subtilis fermented liquid, the number of effective viable bacteria in the bacillus mycoides fermented liquid and the number of effective viable bacteria in the nocardia corallina fermented liquid are all more than or equal to 1011cfu/mL。
Preferably, the carrier is prepared from grass carbon, bamboo charcoal and sodium carboxymethyl cellulose according to the weight ratio of 10: 5: 1, and mixing the components according to the mass ratio.
The second purpose of the invention is to provide a preparation method of a microbial compound inoculant, which comprises the following steps:
step 1, respectively preparing a bacillus subtilis seed solution, a bacillus mycoides seed solution and a nocardiopsis corallina seed solution;
step 2, preparing bacillus subtilis zymocyte liquid
Mixing mulberry leaf, silkworm excrement and fish meal according to the weight ratio of 10: 3: 1, grinding into powder after uniformly mixing, and then sieving by a 80-mesh sieve to obtain mixed raw material powder; mixing the mixed raw material powder with wood vinegar and water according to the weight ratio of 15: 1: 20 to obtain a bacillus subtilis culture medium;
inoculating a bacillus subtilis seed solution into a bacillus subtilis culture medium, uniformly mixing, and then culturing at 28 ℃ at an oscillation speed of 200r/min to the end of logarithm to obtain a bacillus subtilis zymocyte solution;
step 3, preparing a bacillus mycoides fermentation bacteria liquid
Mashing pericarpium Granati into pericarpium Granati mud, adding water 20 times of the total weight of pericarpium Granati mud, soaking for 1-2 hr, and filtering to obtain pericarpium Granati juice; mixing soybean meal powder, soybean molasses, glutamine and pomegranate peel juice according to the weight ratio of 30: 30: 1: 60 to obtain a mushroom bacillus culture medium;
inoculating a bacillus mycoides seed solution into a bacillus mycoides culture medium, uniformly mixing, and then culturing at 28 ℃ at a shaking speed of 200r/min to the end of logarithm to obtain a bacillus mycoides zymocyte solution;
step 4, preparing the rhodobacter corallinus fermentation bacterium liquid
Mixing potato extract powder, yeast extract powder, glucose, diethyl aminoethyl hexanoate and water according to the weight ratio of 30: 15: 40: 1: 100, and obtaining the culture medium of the nocardia corallina;
inoculating a coral nocardia species seed solution into a coral nocardia species culture medium, uniformly mixing, and then culturing at 28 ℃ at an oscillation speed of 200r/min to the late logarithmic phase to obtain a coral nocardia species zymocyte solution;
step 5, weighing the following components in parts by weight: 30-50 parts of bacillus subtilis fermented bacterial liquid, 20-30 parts of bacillus mycoides fermented bacterial liquid, 10-20 parts of corallina nocardia fermented bacterial liquid, 10-20 parts of functional microbial inoculum and 60-80 parts of carrier;
uniformly mixing the weighed bacillus subtilis fermented bacteria liquid, the bacillus mycoides fermented bacteria liquid and the nocardia corallina type bacteria fermented bacteria liquid to obtain a mixed fermented bacteria liquid;
and uniformly mixing the mixed zymogen liquid with the weighed functional microbial inoculum and the carrier, and drying to obtain the microbial compound microbial inoculum.
Preferably, the inoculation amounts of the bacillus subtilis seed solution, the bacillus mycoides fermented solution and the nocardia corallina seed solution are 5-10mL seed solution/kg matrix.
Preferably, the water content of the microbial compound inoculum obtained after drying in the step 5 is 5-10%.
The third purpose of the invention is to provide an application of the microbial compound inoculant as a bacterial fertilizer in crop growth.
In the invention, the bacillus subtilis can decompose silicate substances in soil minerals and release potassium in the soil minerals from mineral lattices to be utilized by crops; the bacillus mycoides has the functions of improving soil granular structure and converting elements such as nitrogen, phosphorus and the like, and the metabolite of the bacillus mycoides has the effects of inhibiting pathogenic bacteria, killing insects and weeding; nocardia corallina has the ability to secrete lactic acid with strong bactericidal ability, inhibit the activity of harmful microorganisms, accelerate the putrefactive decomposition of organic substances, and decompose lignin, cellulose and the like into small molecular substances. The three strains are matched with each other, so that the improvement of the soil performance and the improvement of the soil fertility are promoted. In addition, the functional bacteria can also provide certain nitrogen, phosphorus and potassium for the soil, so that the soil fertility is further improved.
Compared with the prior art, the invention has the beneficial effects that:
1) the invention adopts specific culture medium to culture different strains in a targeted manner, so that the effective viable count in each strain fermentation liquor is more than or equal to 1011cfu/mL can ensure that the effective viable count in the prepared microbial fertilizer reaches the product quality standard; and because the prepared fermentation liquor has high effective colony number, the fermentation liquor can be directly used, and the strain is in a stable environment, thereby being beneficial to the maintenance of the activity of the strain and the exertion of the function.
2) The invention utilizes the synergistic effect of the bacillus subtilis, the bacillus mycoides, the nocardia corallina and the functional microbial inoculum, obviously increases the content of organic matters, nitrogen, phosphorus, potassium and other nutrient elements in the soil, improves the nutrient conversion rate and the number of beneficial microorganisms in the product, promotes the soil remediation, fertilizes the soil fertility, reduces the usage amount of chemical fertilizers, simultaneously enhances the disease resistance of crops, improves the quality of the crops, reduces the investment, improves the yield and promotes the income increase.
3) According to the invention, the turf, the bamboo charcoal and the sodium carboxymethyl cellulose are used as carriers of the microbial composite inoculant, so that microorganisms can be well loaded on the carriers; the turf is a common microorganism loading carrier, but due to the characteristics, the turf is difficult to dry after being loaded with microorganisms, so that the water content of the prepared microbial fertilizer is high, and the effective utilization rate of the microbial fertilizer is influenced; the bamboo charcoal has a large number of micropores, can increase the load rate of microorganisms, but has lower binding capacity to the microorganisms than the grass carbon, so that the bamboo charcoal is matched with the grass carbon to improve the load rate of the microorganisms and the binding force with the microorganisms, and in order to further improve the binding force between the microorganisms, sodium carboxymethyl cellulose is added as an auxiliary additive; in addition, the sodium carboxymethyl cellulose also has a slow release function, so that the microbial fertilizer can slowly and long-term play a role, and the long-term improvement of the soil performance is facilitated.
Detailed Description
In order to make the technical solutions of the present invention better understood and enable those skilled in the art to practice the present invention, the following embodiments are further described, but the present invention is not limited to the following embodiments.
The Bacillus subtilis, Bacillus mycoides and Nocardia corallina strains used in the following examples are all the existing strains which can be purchased in the microorganism strain preservation management center, do not relate to the development of new strains, and only relate to the application of the existing strains. In addition, the bacillus subtilis used in the following examples is specifically a strain with the Chinese agricultural microbial culture collection management center preservation number of ACCC 11089; the bacillus mycoides is a strain with the preservation number of CGMCC No.11047 of China Committee for culture Collection of microorganisms; the Nocardia coralline is specifically a strain with the preservation number of ACCC 40100 of China agricultural microorganism culture preservation management center.
In the examples, the bacillus subtilis seed solution, the bacillus mycoides seed solution, and the nocardiopsis corallina seed solution were obtained by culturing according to a conventional method, the azotobacteria inoculant and rhizobia inoculant used were purchased from pacific concerned (Weifang) Biotech ltd, and the experimental methods described in the following examples are according to a conventional method unless otherwise specified.
Example 1
A composite microbial inoculum is prepared from the following raw materials in parts by weight: 40 parts of bacillus subtilis zymocyte liquid, 25 parts of bacillus mycoides zymocyte liquid, 15 parts of corallina rhodochrous zymocyte liquid, 15 parts of functional microbial inoculum and 70 parts of carrier;
the preparation method comprises the following steps:
step 1, respectively preparing a bacillus subtilis seed solution, a bacillus mycoides seed solution and a nocardiopsis corallina seed solution;
step 2, preparing bacillus subtilis zymocyte liquid
Mixing mulberry leaf, silkworm excrement and fish meal according to the weight ratio of 10: 3: 1, grinding into powder after uniformly mixing, and then sieving by a 80-mesh sieve to obtain mixed raw material powder; mixing the mixed raw material powder with wood vinegar and water according to the weight ratio of 15: 1: 20 to obtain a bacillus subtilis culture medium;
inoculating a bacillus subtilis seed solution into a bacillus subtilis culture medium, uniformly mixing, and then culturing at 28 ℃ at an oscillation speed of 200r/min to the end of logarithm to obtain a bacillus subtilis zymocyte solution;
step 3, preparing a bacillus mycoides fermentation bacteria liquid
Mashing pericarpium Granati into pericarpium Granati mud, adding water 20 times of the total weight of pericarpium Granati mud, soaking for 1-2 hr, and filtering to obtain pericarpium Granati juice; mixing soybean meal powder, soybean molasses, glutamine and pomegranate peel juice according to the weight ratio of 30: 30: 1: 60 to obtain a mushroom bacillus culture medium;
inoculating a bacillus mycoides seed solution into a bacillus mycoides culture medium, uniformly mixing, and then culturing at 28 ℃ at a shaking speed of 200r/min to the end of logarithm to obtain a bacillus mycoides zymocyte solution;
step 4, preparing the rhodobacter corallinus fermentation bacterium liquid
Mixing potato extract powder, yeast extract powder, glucose, diethyl aminoethyl hexanoate and water according to the weight ratio of 30: 15: 40: 1: 100, and obtaining the culture medium of the nocardia corallina;
inoculating a coral nocardia species seed solution into a coral nocardia species culture medium, uniformly mixing, and then culturing at 28 ℃ at an oscillation speed of 200r/min to the late logarithmic phase to obtain a coral nocardia species zymocyte solution;
step 5, weighing the following components in parts by weight: 40 parts of bacillus subtilis zymocyte liquid, 25 parts of bacillus mycoides zymocyte liquid, 15 parts of corallina rhodochrous zymocyte liquid, 15 parts of functional microbial inoculum and 70 parts of carrier;
uniformly mixing the weighed bacillus subtilis fermented bacteria liquid, the bacillus mycoides fermented bacteria liquid and the nocardia corallina type bacteria fermented bacteria liquid to obtain a mixed fermented bacteria liquid;
and (3) uniformly mixing the mixed zymogen liquid with the weighed functional microbial inoculum and the carrier, and drying to obtain the microbial composite microbial inoculum with the water content of 5%.
Example 2
A composite microbial inoculum is prepared from the following raw materials in parts by weight: 50 parts of bacillus subtilis zymocyte liquid, 30 parts of mushroom-shaped bacillus zymocyte liquid, 10 parts of coralline nocardia zymocyte liquid, 10 parts of functional microbial inoculum and 60 parts of carrier.
The preparation method is the same as that of example 1, except that the formula is changed into the formula of the microbial composite inoculum of example 2, and the water content of the prepared microbial composite inoculum is 10%.
Example 3
A composite microbial inoculum is prepared from the following raw materials in parts by weight: 30 parts of bacillus subtilis zymocyte liquid, 20 parts of mushroom-shaped bacillus zymocyte liquid, 20 parts of coralline nocardia zymocyte liquid, 20 parts of functional microbial inoculum and 80 parts of carrier.
The preparation method is the same as that of example 1, except that the formula is changed into the formula of the microbial composite inoculum of example 3, and the water content of the prepared microbial composite inoculum is 8%.
The effective number of viable bacteria in the Bacillus subtilis fermented solution, the effective number of viable bacteria in the Bacillus mycoides fermented solution and the effective number of viable bacteria in the rhodomyces corallinus fermented solution are all equal to or more than 1011cfu/mL；
The functional microbial inoculum is prepared from azotobacter microbial inoculum and rhizobium microbial inoculum according to the ratio of 3: 1, and the effective viable count of the azotobacteria microbial inoculum and the rhizobium microbial inoculum is more than or equal to 1010cfu/g；
The carrier is prepared from grass carbon, bamboo charcoal and sodium carboxymethylcellulose according to the weight ratio of 10: 5: 1, and mixing the components according to the mass ratio.
It is further noted that the inoculum size of the Bacillus subtilis seed solution, the Bacillus mycoides fermented solution and the Nocardia corallina seed solution is 5-10mL seed solution/kg matrix.
In order to further illustrate the effect, the invention is also provided with a comparative example which is concretely as follows.
Comparative example 1
The formula and the preparation method of the microbial compound inoculant are the same as those in example 1, except that a conventional culture medium is adopted in the preparation method for preparing the bacillus subtilis zymocyte liquid, and the specific formula comprises 1L of distilled water, 20g of glucose, 15g of peptone, 5g of sodium chloride, 0.5g of beef extract and 20g of agar;
preparing a bacillus mycoides zymocyte solution by using a conventional culture medium, wherein the specific formula is 1L of distilled water, 10g of yeast extract, 15g of glycerol and 5g of soluble starch;
the preparation of the rhodobacter corallinus zymocyte liquid also adopts a conventional culture medium, and the specific formula is 1L of distilled water, 15g of potato extract powder, 5g of yeast extract powder, 20g of glucose and 10g of animal waste.
Comparative example 2
The formula and the preparation method of the microbial compound inoculum are the same as those in example 1, except that the carrier is turf, and the water content of the finally prepared microbial compound inoculum after drying is 25%.
Comparative example 3
The preparation method of the microbial compound inoculant is the same as that in the example 1, except that the formula does not contain the rhodobacter corallinum hance fermentation broth, and the dosage of other components in the formula is the same as that in the example 1.
The effective viable count of the Bacillus subtilis fermented solution, the Bacillus mycoides fermented solution and the Nocardia corallina fermented solution prepared in examples 1-3 and comparative example 1 was determined, and the specific results are shown in Table 1.
TABLE 1 effective number of viable bacteria in each fermentation broth (10)10cfu/g)
As can be seen from Table 1, the number of effective viable bacteria in each of the fermented bacterial liquids cultured in examples 1-3 is greater than that in comparative example 1, because the present invention adds some components which contribute to the mass and rapid propagation of each strain according to the characteristics of each strain when preparing the culture medium, the number of effective viable bacteria in the fermented bacterial liquid cultured finally is greater than that cultured by using the conventional culture medium.
The effect of the present invention will be further explained with reference to the application of the microbial composite inoculum prepared by the present invention in wheat planting, and the specific test process and results are as follows.
Test subjects: selecting 70-mu test fields in a wheat planting base, dividing the test fields into 7 groups of 10 mu each, wherein three groups are test groups, and applying the microbial compound inoculum prepared in the embodiment 1-3; the other three groups are comparison groups, and the microbial compound inoculum prepared in the comparative examples 1-3 is applied; the last group is a blank control group, and no microbial compound inoculum is added.
It should be noted that, ordinary farmyard manure is applied to 7 groups of test fields before wheat is planted.
The test method comprises the following steps: applying farmyard manure at the same time, and planting wheat of a test group, a comparison group and a blank control group at the same time, wherein the application amount of the farmyard manure is 500 kg/mu, and the application amount of the microbial compound inoculant prepared in the examples 1-3 is 25 kg/mu when the farmyard manure is applied to the test group besides the farmyard manure; the control group also applied the microbial composite inoculum prepared in comparative examples 1-3 while applying farmyard manure in addition to farmyard manure.
It is noted that no compound fertilizer and pesticide are applied to the wheat during the growth process.
In the test, the plough layer soil before applying the fertilizer and after harvesting the wheat is respectively detected, the basic physicochemical values of the plough layer soil are analyzed, and the specific results are shown in table 2.
TABLE 2 results of soil physicochemical Properties test
As can be seen from Table 2, after the microbial compound inoculum of examples 1-3 is applied to the soil of the wheat test field, the physicochemical properties of the soil are greatly improved compared with those of comparative examples 1-3 and a control group, the content of nutrient elements in the soil is greatly increased, the fertility is also greatly improved, the volume weight of the soil is reduced, the porosity of the soil is increased, and the physicochemical properties of the soil are better.
The number of microorganisms in comparative examples 1 and 2 is relatively small, so that the soil has better effect in all aspects
Examples 1-3 are poor, but the difference is not too great; the comparative example 3 has less synergistic effect of the two types of bacteria, so the effect is not as good as that of examples 1-3 and is not as good as that of comparative examples 1-2.
Generally, nitrogen and phosphorus elements contained in farmyard manure are low, and the nitrogen and phosphorus elements are also influenced by soil environment (air, temperature, moisture, microbial activity) and crop vitality and respiration intensity, so that the content can be further reduced, and in order to maintain crop growth, a compound fertilizer is generally required to be additionally applied, and after the microbial compound inoculant is applied in the examples 1-3, on one hand, the soil microenvironment can be improved, and on the other hand, each zymogen liquid and functional bacteria in the microbial compound inoculant can increase the content of nitrogen, phosphorus and potassium in soil, so that the soil fertility is improved, and the requirements of soil microbes and crop growth are met.
After wheat was planted, no other fertilizer was applied during the growth process, and the yield of wheat was recorded after harvesting, and the specific results are shown in table 3.
TABLE 3 wheat yield
| Item | Wheat yield (kg/hm)2) |
| Example 1 | 6578.5 |
| Example 2 | 6583.6 |
| Example 3 | 6610.1 |
| Comparative example 1 | 6219.3 |
| Comparative example 2 | 6486.9 |
| Comparative example 3 | 5936.8 |
| Control group | 5568.7 |
As can be seen from table 3, after the wheat is planted by applying the microbial composite inoculant of example 1, compared with the wheat planted without applying the microbial composite inoculant, the yield of the wheat is improved by 18.13%; after the microbial compound inoculant in the embodiment 2 is applied to plant wheat, compared with the wheat which is not applied with the microbial compound inoculant, the yield of the wheat is improved by 18.22%, and after the microbial compound inoculant in the embodiment 3 is applied to plant the wheat, the yield of the wheat is improved by 18.70% compared with the wheat which is not applied with the microbial compound inoculant; therefore, the microbial compound inoculant disclosed by the invention can improve the physical and chemical properties of soil, improve the soil fertility and further improve the crop yield.
The comparative example 1 and the comparative example 2 have relatively small microbial numbers, so that the effect of the soil in all aspects is poorer than that of the examples 1-3, and the yield of the planted wheat is lower than that of the examples 1-3; in the comparative example 3, the cooperation of the two strains is reduced, the physical and chemical properties of the soil are poorer, and the wheat yield is lower; in addition, since comparative example 3 does not contain Bacillus mycoides and Nocardia corallina, the microbial composite inoculum has substantially no bactericidal and insecticidal functions, thereby further affecting wheat yield.
It should be noted that when the present specification refers to numerical ranges, it should be understood that two endpoints of each numerical range and any value between the two endpoints can be selected, and since the steps and methods adopted are the same as those of the embodiments 1 to 3, the preferred embodiments are described in the present specification for avoiding redundancy, but once the basic inventive concept is known, those skilled in the art can make other changes and modifications to the embodiments. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (7)
1. The microbial compound inoculant is characterized by comprising the following raw materials in parts by weight: 30-50 parts of bacillus subtilis fermented bacterial liquid, 20-30 parts of bacillus mycoides fermented bacterial liquid, 10-20 parts of corallina nocardia fermented bacterial liquid, 10-20 parts of functional microbial inoculum and 60-80 parts of carrier;
the functional microbial inoculum is prepared from azotobacter microbial inoculum and rhizobium microbial inoculum according to the ratio of 3: 1, and the number of effective viable bacteria in the azotobacter and the rhizobium inoculant is more than or equal to 1010cfu/g；
The carrier is prepared from grass carbon, bamboo charcoal and sodium carboxymethyl cellulose according to the weight ratio of 10: 5: 1, mixing and preparing;
the bacillus subtilis is a bacterial strain with the preservation number of ACCC 11089 in China agricultural microbial strain preservation management center; the bacillus mycoides is a bacterial strain with the preservation number of CGMCC No.11047 of China Committee for culture Collection of microorganisms; the nocardia corallina is specifically a strain with the preservation number of ACCC 40100 of China agricultural microorganism strain preservation management center.
2. The microbial composite inoculant according to claim 1, wherein the preparation raw materials comprise the following components in parts by weight: 40 parts of bacillus subtilis zymocyte liquid, 25 parts of mushroom-shaped bacillus zymocyte liquid, 15 parts of coralline nocardia zymocyte liquid, 15 parts of functional microbial inoculum and 70 parts of carrier.
3. The microbial composite inoculant according to claim 1 or 2, wherein the number of active bacteria in the Bacillus subtilis fermented inoculant, the number of active bacteria in the Bacillus mycoides fermented inoculant, and the number of active bacteria in the Nocardia coralline fermented inoculant are all equal to or greater than 1011cfu/mL。
4. The method for preparing a composite microbial inoculum according to claim 1, comprising the steps of:
step 1, respectively preparing a bacillus subtilis seed solution, a bacillus mycoides seed solution and a nocardiopsis corallina seed solution;
step 2, preparing bacillus subtilis zymocyte liquid
Mixing mulberry leaf, silkworm excrement and fish meal according to the weight ratio of 10: 3: 1, grinding into powder after uniformly mixing, and then sieving by a 80-mesh sieve to obtain mixed raw material powder; mixing the mixed raw material powder with wood vinegar and water according to the weight ratio of 15: 1: 20 to obtain a bacillus subtilis culture medium;
inoculating a bacillus subtilis seed solution into a bacillus subtilis culture medium, uniformly mixing, and then culturing at 28 ℃ at an oscillation speed of 200r/min to the end of logarithm to obtain a bacillus subtilis zymocyte solution;
step 3, preparing a bacillus mycoides fermentation bacteria liquid
Mashing pericarpium Granati into pericarpium Granati mud, adding water 20 times of the total weight of pericarpium Granati mud, soaking for 1-2 hr, and filtering to obtain pericarpium Granati juice; mixing soybean meal powder, soybean molasses, glutamine and pomegranate peel juice according to the weight ratio of 30: 30: 1: 60 to obtain a mushroom bacillus culture medium;
inoculating a bacillus mycoides seed solution into a bacillus mycoides culture medium, uniformly mixing, and then culturing at 28 ℃ at a shaking speed of 200r/min to the end of logarithm to obtain a bacillus mycoides zymocyte solution;
step 4, preparing the rhodobacter corallinus fermentation bacterium liquid
Mixing potato extract powder, yeast extract powder, glucose, diethyl aminoethyl hexanoate and water according to the weight ratio of 30: 15: 40: 1: 100, and obtaining the culture medium of the nocardia corallina;
inoculating a coral nocardia species seed solution into a coral nocardia species culture medium, uniformly mixing, and then culturing at 28 ℃ at an oscillation speed of 200r/min to the late logarithmic phase to obtain a coral nocardia species zymocyte solution;
step 5, weighing the following components in parts by weight: 30-50 parts of bacillus subtilis fermented bacterial liquid, 20-30 parts of bacillus mycoides fermented bacterial liquid, 10-20 parts of corallina nocardia fermented bacterial liquid, 10-20 parts of functional microbial inoculum and 60-80 parts of carrier;
uniformly mixing the weighed bacillus subtilis fermented bacteria liquid, the bacillus mycoides fermented bacteria liquid and the nocardia corallina type bacteria fermented bacteria liquid to obtain a mixed fermented bacteria liquid;
and uniformly mixing the mixed zymogen liquid with the weighed functional microbial inoculum and the carrier, and drying to obtain the microbial compound microbial inoculum.
5. The method for preparing a composite microbial inoculum according to claim 4, wherein the inoculum size of the Bacillus subtilis seed solution, the Bacillus mycoides fermented solution and the Nocardia corallina seed solution is 5-10mL seed solution/kg substrate.
6. The method for preparing a composite microbial inoculum according to claim 4, wherein the water content of the composite microbial inoculum obtained after drying in the step 5 is 5-10%.
7. The application of the microbial compound inoculant of claim 1 as a bacterial fertilizer in crop growth.
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| CN110257074A (en) * | 2019-04-01 | 2019-09-20 | 谷峻峰 | A kind of preparation of wood vinegar bacillus subtilis bacterium solution soil conditioner and its application method |
| CN113150788A (en) * | 2021-04-22 | 2021-07-23 | 浙江树人学院（浙江树人大学） | Preparation method and application of seaweed oligosaccharide composite microbial material for preventing soil hardening |
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