CN111334456A - Preparation method and application of straw decomposition microbial inoculum used under low-temperature condition in north - Google Patents
Preparation method and application of straw decomposition microbial inoculum used under low-temperature condition in north Download PDFInfo
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Abstract
The invention relates to a preparation method of a straw decomposition microbial inoculum used in the northern low-temperature condition, which is characterized by comprising the following steps: respectively and independently culturing functional bacteria of bacillus subtilis, trichoderma longibrachiatum and myceliophthora thermophila in a liquid basal culture medium to prepare a primary seed fermentation liquid, then performing secondary fermentation propagation in a secondary fermentation culture medium to prepare a secondary fermentation liquid, and performing secondary fermentation propagation on the three types of propagated secondary fermentation liquids according to a ratio of 1:1:1 to obtain a liquid decomposition microbial inoculum; the method utilizes low-temperature functional bacteria obtained by screening to prepare the crop straw decomposition microbial inoculum and verifies the decomposition effect below zero DEG C at the northern outdoor temperature, and belongs to the field of plant nutrition and land sustainable utilization in agricultural production practice.
Description
Technical Field
The invention relates to a preparation method and application of a straw decomposition microbial inoculum under the low-temperature condition in the north, which utilizes low-temperature functional bacteria obtained by screening to prepare a crop straw decomposition microbial inoculum and verifies the decomposition effect below zero DEG C in the outdoor temperature in the north, and belongs to the field of plant nutrition and sustainable utilization of land in agricultural production practice.
Background
The straw as high-quality biomass energy can partially replace and save fossil energy, and the dependence on the fossil energy is reduced. According to heat value measurement, 2 tons of straws are equivalent to 1 ton of standard coal, straw energy is developed and utilized, energy supply in rural areas can be effectively increased, energy structure is improved, and carbon dioxide emission is reduced. And secondly, the straw contains rich organic matters, nitrogen, phosphorus, potassium and trace elements, is a multipurpose and renewable biological resource, and is also an important organic fertilizer source for agricultural production. According to measurement and calculation, 350 million tons of nitrogen, 80 million tons of phosphorus and 800 million tons of potassium are contained in 7 million tons of straws, which is about one fifth of the total amount of the chemical fertilizer applied nationwide in 2010. And thirdly, the straw fiber is a natural cellulose fiber, has good biodegradability, can replace wood to be used for papermaking, producing boards, manufacturing artware, producing activated carbon and the like, saves a large amount of wood, and protects valuable forest resources. And fourthly, the straws contain rich nutrient substances, the nutrient value of 4 tons of straws is equivalent to that of 1 ton of grains, and the material guarantee can be provided for the continuous development of the animal husbandry. This provides very important theoretical significance for the comprehensive utilization of the straw.
The main active ingredients of the straw decomposition microbial inoculum are prepared from aspergillus niger, trichoderma pseudokoningii, fresh penicillium viride, clostridium xylanilyticum, pseudomonas and bacillus subtilis. The effect of the fertilizer can lead the decomposition rate of the wheat and rice straws to reach 60.1 percent in 15 days.
The invention relates to a straw decomposition agent and a preparation and an application thereof, which are characterized in that pectin, polysaccharide-polysaccharide compound and lignin polysaccharide compound existing in straw cell walls can be effectively degraded to form free polysaccharide, and the degree and the speed of further degradation are increased; the preparation method of the decomposition agent is simple and practical, the microbial inoculum has good activity and high enzyme yield, and the prepared straw organic fertilizer has rich nutrient substances and effectively improves the soil condition.
Disclosure of Invention
The invention aims to provide a preparation method and application of a straw decomposition microbial inoculum used under the low-temperature condition in the north, which can promote the rapid decomposition of crop straws under the low-temperature condition in the north, simultaneously can improve the physical properties of soil, increase the contents of organic matters and quick-acting nutrients in the soil, realize the reutilization of crop straw resources, and have very obvious economic benefit, social benefit and environmental benefit.
The straw decomposition agent contains a large amount of bacteria, mould, saccharomycetes, bacillus and the like, is a high-efficiency microbial preparation, can effectively decompose crop straws by mass propagation of microorganisms in the straw decomposition agent, has the functions of improving the soil granular structure, improving the soil air permeability and fertilizer and water retention and improving the soil nutrient condition, and can improve the plant growth environment and promote the effective utilization of the straws by heat generated by microbial activity and a certain amount of carbon dioxide; therefore, screening and separating high-enzyme activity strains which are suitable for efficiently and quickly degrading cellulase and lignin under the conditions of low temperature and drought climate in the north, and carrying out research on low-temperature quick conversion microbial agent and enzyme preparation of straws and field application regulation and control technology are important scientific problems for solving comprehensive utilization of main crop straws in the northeast of China.
The technical scheme of the invention is realized as follows: a preparation method of a straw decomposition microbial inoculum used in the northern low-temperature condition is characterized by comprising the following steps: respectively and independently culturing functional bacteria of bacillus subtilis, trichoderma longibrachiatum and myceliophthora thermophila in a liquid basal culture medium to prepare a primary seed fermentation liquid, then performing secondary fermentation propagation in a secondary fermentation culture medium to prepare a secondary fermentation liquid, and performing secondary fermentation propagation on the three types of propagated secondary fermentation liquids according to a ratio of 1:1:1 to obtain a liquid decomposition microbial inoculum; the method comprises the following specific steps:
1. first order seed liquid preparation
Taking the activated slant strain 1 ring by using an inoculating ring, inoculating the slant strain 1 ring into a triangular flask containing 100-150 mL of a basic culture medium, and carrying out shaking culture at the temperature of 28-37 ℃ and the pH value of 5.8-7.2 for 24-36 hours at 150-180 r/min to obtain a fermentation broth, namely a first-stage seed fermentation broth;
respectively preparing liquid seed synthetic culture media of bacteria (LB) and fungi (PDB), then placing the liquid culture media into an autoclave for sterilization at 121 ℃ for 30 min, respectively inoculating bacillus subtilis into the LB culture media after cooling, inoculating trichoderma longibrachiatum and myceliophthora thermophila into the PDB culture media, setting different fermentation parameters according to the fermentation processes of the bacteria and the fungi, and obtaining liquid seeds after fermentation culture, namely primary seed liquid;
TABLE 1 preparation of first-class seed liquid culture Medium formulation and culture conditions
2. Preparation of secondary seed fermentation broth
Inoculating the first-stage seed liquid after shaking culture into a fermentation culture medium by 0.5% of inoculation amount, and performing secondary fermentation culture (propagation culture) in large-scale fermentation equipment to obtain a second-stage seed culture liquid, wherein the formula and fermentation conditions of the second-stage fermentation culture medium are shown in Table 2;
TABLE 2 preparation of secondary seed liquid culture medium formula and culture conditions
The culture medium selected by the secondary fermentation liquid is the waste substance of a corn processing factory, namely corn steep liquor, serving as a nitrogen source to replace fermentation mother powder or beef extract, and the cost can be saved by more than 50% in production application;
3. preparation of liquid composite decomposing inoculant
And mixing the fermented secondary seed liquids of the bacillus subtilis, the trichoderma longibrachiatum and the myceliophthora thermophila according to the ratio of 1:1:1 to obtain the liquid compound decomposing inoculant.
The straw decomposition microbial inoculum is applied to decomposing straws of corns and rice under the low-temperature condition in the north.
The invention has the advantages of promoting the fast decomposition of crop straws under the low temperature condition in the north, improving the physical properties of soil and increasing the contents of organic matters and quick-acting nutrients in the soil. 1. Temperature change in the process of stacking straw decomposition microbial inoculum
1.1 decomposition Effect of Single functional Strain on corn stalks
In order to verify the decomposition effect of the straw decomposition bacteria on the corn straws, a cow dung and straw stacking decomposition test is arranged in the field in 3 months in 2018, and 3 treatments are set in the test, namely control treatment without inoculation, trichoderma longibrachiatum and myceliophthora thermophila. The temperature change condition is continuously measured for 11 days, and the result shows that the temperature change difference of different microbial inoculum is obvious, the temperature of the inoculated trichoderma longibrachiatum is continuously stabilized above 40 ℃ on the 5 th day, and the temperature of the myceliophthora thermophila reaches 40 ℃ on the 7 th day. The control temperature without inoculation is always around 20 ℃. The temperature during the measurement is 1 ℃ day, and the temperature during the whole test period is-4 ℃ to 10 ℃ (figure 1).
1.2 decomposition effects of composite functional bacteria on corn and rice straw
In 2019, 6 and 1, a composting test of the straw decomposition microbial inoculum on the decomposition effect of the straws of the corns and the paddy rice is carried out in Guo county before Jilin province, and the fixed time every day is 9 in the morning: 00 continuously monitors the temperature change of the heap corruption process. The results show that the temperature of 6 treatments on day 2 of heap decay has risen above 45 ℃, and the test temperature of the treatments inoculated with the decay microbial inoculum is lower than that of the treatments not inoculated with the microbial inoculum. But the treatment temperature of inoculating the microbial inoculum with different dosage forms by the rice straws is increased to over 58 ℃ by the 5 th day, the treatment temperature of inoculating the solid microbial inoculum by the corn straws reaches 65 ℃, and the treatment temperature of inoculating the liquid microbial inoculum also reaches 63 ℃. Then, along with the prolonging of the fermentation time, the temperature of the compost is maintained to fluctuate at about 60 ℃, the temperature does not show a descending trend until the fermentation lasts for 33 days, and the rotting rate of the corn and the rice straws reaches more than 80 percent (fig. 2 and fig. 3).
1.3 decomposition effect of composite functional bacteria on corn and rice straw at low temperature
11/2019, a composting test of the straw decomposition effect of the straw decomposition microbial inoculum on corn and rice straws is carried out in Helongtown of Changchun city, Jilin province, and the fixed time every day is 7 in the morning: 00 continuously monitors the temperature change of the heap corruption process. The result shows that the temperature of the treatment of inoculating the decomposition bacteria agent by matching the corn and the rice straws with the cow dung on the 2 nd day of the heap rot is increased to more than 50 ℃, and then the temperature is maintained to fluctuate up and down. Particularly, the temperature of the liquid and solid microbial inoculum for inoculation of the rice straws is obviously higher than that of the control treatment without inoculation. And at the moment, the positive value is in severe winter, the outdoor temperature reaches-18 ℃ at the lowest, which shows that the product can survive under the northern low-temperature condition and has stronger activity (figure 4 and figure 5).
2. Influence of straw decomposition microbial inoculum on degradation rate of corn straw
Different kinds of straw decomposition microbial inoculum are applied under the corn straw deep-turning and rotary tillage modes respectively, and the corn straw decomposition rate is measured by sampling at the key phenological stage of the corn. The result shows that the decomposition rate of the inoculated decomposition microbial inoculum of the corn straws in the deep ploughing returning mode is 85.5 percent, which is 7.82 percent higher than that of the products sold in the market and is 8.16 percent higher than that of the straws without the inoculated microbial inoculum. The decomposition rate of the corn straws inoculated with the decomposition microbial inoculum in the rotary tillage returning mode reaches 94.05 percent, is improved by 17.01 percent compared with the straw treatment without the decomposition microbial inoculum, and is improved by 3.18 percent compared with the products sold in the market. The decomposition effect of the product is also influenced by environmental conditions to a great extent besides the influence of the formula. This is also a disadvantage of the biologicals product, since it is a living preparation itself, which is very sensitive to environmental conditions (fig. 6).
3. Influence of soil nutrient content by applying straw decomposition microbial inoculum
3.1 Effect of straw decomposition inoculum on soil available nutrients
3.1.1 Effect of straw decomposition microbial inoculum on content of nitrate nitrogen and ammonium nitrogen in soil
The research result of the influence of the application of the decomposition microbial inoculum on the soil nitrate nitrogen in different straw returning modes shows that the soil nitrate nitrogen content of the application of the decomposition microbial inoculum in the deep ploughing mode is higher than that of the like products on the market and the treatment without applying the microbial inoculum by singly applying the straw, but the improvement range is not very large. The content of soil nitrate nitrogen is improved by 3.8% compared with the similar products in the market by applying the decomposition microbial inoculum under the rotary tillage mode, and is improved by 8.79% compared with the method of applying the microbial inoculum by only applying straws, and the decomposition microbial inoculum also has better effect. The content of soil nitrate nitrogen is improved by 20.29% compared with the similar products in the market and 30.28% compared with the method of applying the straw without the bactericide singly in a mixing mode, and the decomposition bactericide shows excellent effect. Fully embodies the function of the product for improving the content of nitrate nitrogen nutrients in soil (figure 7).
The research result of the influence of applying the decomposition microbial inoculum to the soil ammonium nitrogen in different straw returning modes shows that the content of the soil ammonium nitrogen in the decomposition microbial inoculum applied in the deep ploughing mode is higher than 6.16% of that of like products on the market, and is improved by 8.63% compared with the treatment of applying the microbial inoculum only without applying the straw. The content of the soil ammonium nitrogen by applying the decomposition microbial inoculum under the rotary tillage mode is not greatly improved compared with the similar products in the market, but is improved by 12.21 percent compared with the soil without applying the microbial inoculum by applying straws singly. The content of the soil ammonium nitrogen applied with the decomposition bacteria agent in the mixing mode is improved by 2.61 percent compared with the similar products in the market and is improved by 21.23 percent compared with the soil without applying the bacteria agent by singly applying the straws. The positive effect of the decay microbial inoculum on improving the content of the ammonium nitrogen in the soil is shown (figure 8).
3.1.2 influence of straw decomposition microbial inoculum on content of quick-acting phosphorus and quick-acting potassium in soil
The research result of the influence of the field application of the decomposition microbial inoculum on the soil available phosphorus content in different straw returning modes shows that the soil available phosphorus content is improved by 3.23% when the decomposition microbial inoculum is applied in the deep ploughing mode compared with the like products in the market, and is improved by 5.83% when the decomposition microbial inoculum is applied in the field in a single straw treatment mode. The quick-acting phosphorus content of the soil applied with the decomposition microbial inoculum under the rotary tillage mode is improved by 4.25 percent compared with the similar products in the market and is improved by 9.63 percent compared with the soil applied with straw without the microbial inoculum. The content of the quick-acting phosphorus in the soil applied with the decomposition microbial inoculum in the mixing mode is improved by 6.59 percent compared with the similar products in the market and is improved by 19.15 percent compared with the soil applied with straw without the microbial inoculum. The effect of the decay microbial inoculum on improving the soil nutrient content is illustrated again (fig. 9).
The research result of the influence of field application of the decomposition microbial inoculum on the soil quick-acting potassium content in different straw return modes shows that the soil quick-acting potassium content is improved by 4.98% when the decomposition microbial inoculum is applied in the deep ploughing mode compared with the similar products in the market, and is improved by 6.20% when compared with the single straw application treatment. The quick-acting potassium content of the soil applied with the decomposition microbial inoculum under the rotary tillage mode is improved by 2.63 percent compared with the similar products in the market and is improved by 4.20 percent compared with the soil applied with straw without the microbial inoculum. The content of the quick-acting potassium in the soil applied with the decomposition microbial inoculum under the mixing mode is improved by 3.96 percent compared with the similar products in the market and is improved by 6.06 percent compared with the soil applied with straw without the microbial inoculum. The decomposition microbial inoculum also has positive promoting effect on improving the content of the quick-acting potassium in the soil (figure 10).
3.1.3 Effect of applying straw decomposition microbial inoculum on organic matter content of soil
The research result of the influence of the application of the decomposition microbial inoculum on the content of the organic matters in the soil in different straw returning modes shows that the content of the organic matters in the soil is increased by 16.92 percent compared with the similar products in the market in the deep ploughing mode, and is increased by 9.89 percent compared with the single straw application treatment. The organic matter content of the soil applied with the decomposition microbial inoculum under the rotary tillage mode is improved by 15.01 percent compared with the similar products in the market and is improved by 3.92 percent compared with the soil applied with straw without the microbial inoculum. The organic matter content of the soil applied with the decomposition microbial inoculum under the mixing mode is improved by 10.07 percent compared with the similar products in the market and is improved by 5.77 percent compared with the soil applied with straw without the microbial inoculum. The decomposing bacteria agent has a particularly obvious effect on improving the organic matter content of soil, which is also the innovation of the patent. (FIG. 11).
4. Influence of application of straw decomposition microbial inoculum on biomass and yield of corn
4.1 Effect of straw-decomposing inoculant on corn Biomass
The research result of the influence of applying the decomposition microbial inoculum to the corn biomass in different straw returning modes shows that the corn biomass is improved by 8.11% compared with the similar products in the market in the deep-turning mode, and is improved by 14.67% compared with the single straw application treatment. The corn biomass is increased by 8.95% compared with the similar products in the market and is increased by 9.87% compared with the method of applying straw without applying bacteria in a rotary tillage mode. The biomass of the corn which is applied with the decay microbial inoculum under the mixing mode is improved by 1.24 percent compared with the similar products in the market, and is improved by 8.35 percent compared with the mode that the straw is applied singly without applying the microbial inoculum. The application of the decay microbial inoculum is also shown to have a promoting effect on increasing the biomass of the corn plants (figure 12).
4.2 influence of application of straw-decomposing inoculant on corn yield
The research result of the influence of applying the decomposition microbial inoculum to the corn yield in different straw returning modes shows that the corn biomass is improved by 4.27% compared with the similar products in the market in the deep-turning mode, and is improved by 4.63% compared with the single straw application treatment. The biomass of the corn applying the decay microbial inoculum under the rotary tillage mode is improved by 7.84 percent compared with the similar products in the market and is improved by 8.82 percent compared with the method of applying straw without the microbial inoculum. The biomass of the corn which is applied with the decay microbial inoculum under the mixing mode is improved by 9.59 percent compared with the similar products in the market and is improved by 14.7 percent compared with the corn which is applied with straw singly and is not applied with the microbial inoculum. The application of the decay microbial inoculum can obviously improve the corn yield (figure 13).
Drawings
FIG. 1 is a schematic diagram showing the temperature change of the decomposition process of functional bacteria inoculated with corn straws in different dosage forms.
FIG. 2 is a schematic diagram showing the temperature change during the composting process of corn and rice straw.
FIG. 3 is a decomposition effect diagram of 33 days of corn and rice composting.
FIG. 4 is a schematic diagram showing the temperature change of rice straw in the low-temperature composting process.
FIG. 5 is a schematic diagram of temperature change in a low-temperature corn stalk composting process.
FIG. 6 is a schematic diagram of the decomposition rate of corn straw inoculated with decomposition microbial inoculum in deep ploughing and rotary tillage returning modes.
FIG. 7 is a schematic diagram showing the influence of applying a decomposition microbial inoculum on the nitrate nitrogen content of soil under different straw returning modes.
FIG. 8 is a schematic diagram showing the influence of applying a decomposition microbial inoculum on the content of ammonium nitrogen in soil under different straw returning modes.
FIG. 9 is a schematic diagram showing the influence of field application of a decomposition microbial inoculum on the content of available phosphorus in soil in different modes of straw reduction.
FIG. 10 is a schematic diagram showing the influence of field application of a decomposition microbial inoculum on the content of soil available potassium in different modes of straw reduction.
FIG. 11 is a schematic diagram showing the effect of applying a straw decomposition microbial inoculum on the organic matter content of soil in different straw returning modes.
FIG. 12 is a schematic diagram showing the effect of applying a decomposition microbial inoculum on corn biomass in different straw returning modes.
FIG. 13 is a schematic diagram showing the effect of applying a decomposition microbial inoculum on the yield of corn in different straw returning modes. Detailed Description
The invention is further described with reference to the following examples:
example 1
Preparation of primary seed fermentation broth of bacteria (bacillus subtilis): preparing 100 mL of a bacterial (LB) liquid synthetic culture medium (10 g/L of peptone, 5 g/L of yeast powder and 5 g/L, pH 7.0.0 of sodium chloride) and placing the bacterial (LB) liquid synthetic culture medium into a 250mL triangular flask, then placing the liquid culture medium into an autoclave for sterilization at 121 ℃ for 30 min, respectively inoculating activated bacillus subtilis stored on an inclined plane after cooling, and carrying out shaking culture at 28-35 ℃ and 150-180 r/min for 24-36 h to obtain a first-level seed fermentation liquid.
Example 2
Preparation of bacterial (Bacillus subtilis) secondary seed fermentation broth: preparing a bacterial liquid synthetic culture medium (20 g/L of glucose, 10 ml/L of corn steep liquor, 0.2 g/L of magnesium sulfate, 0.5 g/L of dipotassium phosphate and 0.1 g/L, pH 6.8.8-7.2) and putting the bacterial liquid synthetic culture medium into a large-scale fermentation device, carrying out autoclaving at 121 ℃ for 30 min, cooling the bacterial liquid synthetic culture medium, inoculating the bacterial liquid synthetic culture medium into the primary seed fermentation liquid described in the embodiment 1 according to the inoculation amount of 0.5%, carrying out secondary fermentation culture (propagation culture) and carrying out culture at 28-35 ℃ and 150-180 r/min for 24-36 h to obtain the secondary seed fermentation liquid.
Example 3
Preparation of primary seed fermentation liquor of fungus (trichoderma longibrachiatum): 100 mL of a fungus (PDB) liquid potato synthetic culture medium (potato extract powder is 5 g/L, and glucose is 20 g/L, pH 5.8.5.8-6.2) is prepared and contained in a 250mL triangular flask, then the liquid culture medium is put into an autoclave for sterilization at 121 ℃ for 30 min, after cooling, trichoderma longibrachiatum stored on an activated inclined plane is respectively inoculated, and shaking culture is carried out at 32-37 ℃ and 150-180 r/min for 24-36 h, so as to obtain a first-grade seed fermentation liquid.
Example 4
Preparation of secondary seed fermentation liquor of fungus (trichoderma longibrachiatum): preparing a fungus liquid fermentation culture medium (15 g/L of glucose, 5 g/L of brown sugar, 10 ml/L of corn steep liquor, 0.2 g/L of magnesium sulfate, 0.5 g/L of dipotassium hydrogen phosphate and 801 g/L, pH 5.8.8-6.2 of Tween), putting the fungus liquid fermentation culture medium into large-scale fermentation equipment, carrying out autoclaving at 121 ℃ for 30 min, cooling the fungus liquid fermentation culture medium, inoculating the fungus liquid fermentation medium into the primary seed fermentation liquid according to the inoculation amount of 0.5% to carry out secondary fermentation culture (propagation culture) at 32-37 ℃ and 150-180 r/min for 24-36 h, and obtaining secondary seed fermentation liquid. The culture medium selected by the secondary fermentation liquor is a formula which is applied less in the current production, and is characterized in that the waste substance of a corn processing factory, namely corn steep liquor, is used as a nitrogen source to replace fermentation mother powder or beef extract, and the cost can be saved by more than 50% in the production application.
Example 5
Preparation of primary seed fermentation broth of fungus (myceliophthora thermophila): preparing 100 mL of fungus (PDB) liquid potato synthetic culture medium (5 g/L of potato extract powder and 20 g/L, pH 5.8.8-6.2 g of glucose) and placing the culture medium in a 250mL triangular flask, then placing the liquid culture medium in an autoclave for sterilization at 121 ℃ for 30 min, cooling, respectively inoculating myceliophthora thermophila preserved on an activated inclined plane, and carrying out shaking culture at 32-37 ℃ and 150-180 r/min for 24-36 h to obtain first-level seed fermentation liquor.
Example 6
Preparation of secondary seed fermentation liquor of fungus (myceliophthora thermophila): preparing a fungus liquid fermentation culture medium (15 g/L of glucose, 5 g/L of brown sugar, 10 ml/L of corn steep liquor, 0.2 g/L of magnesium sulfate, 0.5 g/L of dipotassium hydrogen phosphate, 801 g/L of tween and pH 5.8-6.2), putting the fungus liquid fermentation culture medium into a large-scale fermentation device, carrying out autoclaving at 121 ℃ for 30 min, cooling the fungus liquid fermentation culture medium, inoculating 0.5% of inoculum size into the primary seed fermentation liquid described in the embodiment 3, carrying out secondary fermentation culture (propagation culture) at 32-37 ℃ and 150-180 r/min, and culturing for 24-36 h to obtain a secondary seed fermentation liquid. The culture medium selected by the secondary fermentation liquor is a formula which is applied less in the current production, and is characterized in that the waste substance of a corn processing factory, namely corn steep liquor, is used as a nitrogen source to replace fermentation mother powder or beef extract, and the cost can be saved by more than 50% in the production application.
Example 7
Preparing a liquid composite decomposing inoculant: and mixing the secondary seed liquids described in the embodiments 2, 4 and 6 according to the ratio of 1:1:1 to obtain the liquid compound decomposing inoculant. The technical indexes of the liquid compound decomposing inoculant meet the requirements of table 3.
TABLE 3 technical indexes of liquid complex microbial inoculum
Example 8
The composting time and the use method of the decay microbial inoculum are as follows: the northern corn is composted about 9 months later and 10 months earlier after harvest, and is used before inoculation in the second year after one winter decomposition. The method is to stack the crushed straws and the livestock and poultry manure according to the volume ratio of 1: 1. Every 15 cm-25 cm of the thickness is 1 layer, and the straw, the decomposed microbial inoculum, the urea and the livestock and poultry manure are stacked in sequence.
The technical parameters are as follows: (1) the water content of the whole pile of materials is controlled between 60 percent and 65 percent. (2) The height of the heap rot is between 2 m and 2.5 m. (3) The dosage of the decomposed inoculant is 0.25 percent of the weight of the straws, and the dosage of the urea is 0.5 percent of the weight of the straws.
Example 9
The temperature change in the process of decomposing the corn straws by the single functional strain is as follows: cow dung and straw composting tests are arranged in the field in 3 months in 2018, and 3 treatments, control treatment without inoculation, trichoderma longibrachiatum and myceliophthora thermophila are set in the tests. The temperature change condition is continuously measured for 11 days, and the result shows that the temperature change difference of different microbial inoculum is obvious, the temperature of the inoculated trichoderma longibrachiatum is continuously stabilized above 40 ℃ on the 5 th day, and the temperature of the myceliophthora thermophila reaches 40 ℃ on the 7 th day. The control temperature without inoculation is always around 20 ℃. The temperature during the measurement is 1 ℃ day, and the temperature during the whole test period is-4 ℃ to 10 ℃ (figure 1).
Example 10
The temperature effect of the composite functional bacteria in the process of decomposing the corn and the rice straws is as follows: 11/2019, a composting test of the straw decomposition effect of the straw decomposition microbial inoculum on corn and rice straws is carried out in Helongtown of Changchun city, Jilin province, and the fixed time every day is 7 in the morning: 00 continuously monitors the temperature change of the heap corruption process. The result shows that the temperature of the treatment of inoculating the decomposition bacteria agent by matching the corn and the rice straws with the cow dung on the 2 nd day of the heap rot is increased to more than 50 ℃, and then the temperature is maintained to fluctuate up and down. Particularly, the temperature of the liquid and solid microbial inoculum for inoculation of the rice straws is obviously higher than that of the control treatment without inoculation. And at the moment, the positive value is in severe winter, the outdoor temperature reaches-18 ℃ at the lowest, which shows that the product can survive under the northern low-temperature condition and has stronger activity (figure 4 and figure 5).
Claims (2)
1. The preparation method of the straw decomposition microbial inoculum used in the northern low-temperature condition is characterized in that: respectively and independently culturing functional bacteria of bacillus subtilis, trichoderma longibrachiatum and myceliophthora thermophila in a liquid basal culture medium to prepare a primary seed fermentation liquid, then performing secondary fermentation propagation in a secondary fermentation culture medium to prepare a secondary fermentation liquid, and performing secondary fermentation propagation on the three types of propagated secondary fermentation liquids according to a ratio of 1:1:1 to obtain a liquid decomposition microbial inoculum; the method comprises the following specific steps:
1) first order seed liquid preparation
Taking the activated slant strain 1 ring by using an inoculating ring, inoculating the slant strain 1 ring into a triangular flask containing 100-150 mL of a basic culture medium, and carrying out shaking culture at the temperature of 28-37 ℃ and the pH value of 5.8-7.2 for 24-36 hours at 150-180 r/min to obtain a fermentation broth, namely a first-stage seed fermentation broth;
respectively preparing liquid seed synthetic culture media of bacteria (LB) and fungi (PDB), then placing the liquid culture media into an autoclave for sterilization at 121 ℃ for 30 min, respectively inoculating bacillus subtilis into the LB culture media after cooling, inoculating trichoderma longibrachiatum and myceliophthora thermophila into the PDB culture media, setting different fermentation parameters according to the fermentation processes of the bacteria and the fungi, and obtaining liquid seeds after fermentation culture, namely primary seed liquid; the formula and culture conditions for preparing the first-class seed liquid culture medium are as follows:
2) preparation of secondary seed fermentation broth
The culture medium selected by the secondary fermentation liquid is prepared by using corn steep liquor which is waste material of a corn processing factory as a nitrogen source to replace fermentation mother powder or beef extract, and the preparation method comprises the following steps:
inoculating the first-stage seed liquid after shaking culture into a fermentation culture medium by 0.5 percent of inoculation amount, performing secondary fermentation culture in large-scale fermentation equipment, and performing propagation culture to obtain a second-stage seed culture liquid, wherein the formula of the second-stage fermentation culture medium is as follows:
3) preparation of liquid composite decomposing inoculant
And mixing the fermented secondary seed liquids of the bacillus subtilis, the trichoderma longibrachiatum and the myceliophthora thermophila according to the ratio of 1:1:1 to obtain the liquid compound decomposing inoculant.
2. The straw decomposition microbial inoculum is applied to decomposing straws of corns and rice under the low-temperature condition in the north.
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