CN113755342A - Low-temperature-resistant straw fermentation inoculant and preparation method thereof - Google Patents
Low-temperature-resistant straw fermentation inoculant and preparation method thereof Download PDFInfo
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- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
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Abstract
The invention discloses a low-temperature-resistant straw fermentation microbial inoculum and a preparation method thereof, belonging to the technical field of fermentation microbial inoculants. Aiming at solving the technical problem of incomplete straw fermentation in low-temperature environment. The invention provides a straw fermentation inoculant which comprises the following components: saccharomyces cerevisiae, lactobacillus, Bacillus subtilis, photosynthetic bacteria, actinomycetes and Trichoderma koningii. The straw fermentation inoculant provided by the invention can complete straw composting fermentation at a low temperature of-20-15 ℃, can complete fermentation in about 2-3 months under the low temperature conditions of spring and autumn, and is simple to operate and low in cost: the rice straw is thoroughly decomposed, and the operation can be carried out in the field without crushing or building a fermentation tank.
Description
Technical Field
The invention belongs to the technical field of fermentation inoculants, and particularly relates to a low-temperature-resistant straw fermentation inoculant and a preparation method thereof.
Background
The northeast is the largest corn producing area in China, and the straw yield is huge. The burning of the straws in late autumn and early spring is the most common and simple means for farmers to treat the straws. The proportion of direct returning of the corn straws to the field in the region is only 30.9 percent, which is far lower than the average level in China. A large amount of straw resources are discarded or incinerated, which brings environmental problems and simultaneously causes waste of resources.
The straw decomposing and returning to the field is after harvesting of autumn corns in the northeast region, because the temperature drop speed is high, the straw decomposing bacteria applied to the field at present can not complete fermentation under the low-temperature condition, and the cultivation and production of the next year are directly influenced, so that the low-temperature-resistant microbial degradation bacteria need to be screened urgently, the low-temperature-resistant microbial straw fermentation microbial inoculum is developed, the rapid degradation of the straws is promoted efficiently, the problem that the straws treated after the crops are harvested return to the field in time when the straws are sowed in the spring of the next year is solved, the resource recycling of the straws is realized, the agricultural ecological environment is protected, and the sustainable development of modern agriculture is promoted. The rapid degradation of the straws can be efficiently promoted through the action of microorganisms, the popularization significance of accelerating the straw returning is great, and the method is an effective method for reasonably and effectively utilizing straw resources at present.
The variety and activity of the straw degrading bacteria are the most important core technology for determining the decomposition speed and the decomposition degree of the straw under the low-temperature condition. The currently widely used straw degradation microbial inoculum has the disadvantages of low temperature resistance, weak activity under low temperature conditions, slow straw fermentation, incomplete decomposition and incomplete preparation extraction and preparation process. The fermentation time is half a year to more than one year, and the cultivation and the crop growth in the next year are directly influenced. Therefore, low temperature resistant microbial degradation bacteria need to be screened, low temperature resistant microbial straw fermentation inoculants are developed, the rapid degradation of the straws is efficiently promoted, the problem that the straws treated after the crops are harvested return to the ground in time when the straws are sowed in the spring in the next year is solved, the resource recycling of the straws is realized, the agricultural ecological environment is protected, and the sustainable development of modern agriculture is promoted.
Disclosure of Invention
The invention aims to solve the technical problem of incomplete straw fermentation in a low-temperature environment.
The invention provides a low-temperature-resistant straw fermentation microbial inoculum, which consists of the following components: saccharomyces cerevisiae (Saccharomyces cerevisiae), lactic acid Bacteria (Lactobacillus), Bacillus subtilis (Bacillus subtilis), Photosynthetic Bacteria (Photosyntetic Bacteria), Actinomycetes (Actinomycete), Trichoderma koningii (Trichoderma koningii).
Further defined, the straw fermentation inoculum also comprises livestock and poultry manure and straw.
Further limiting, the mass ratio of the straws to the animal manure is 2: 1.
Further limiting, the moisture of the straw is adjusted to 65%.
Further limited, the stalk fermentation inoculum consists of the following mass proportions: brewing yeast: lactic acid bacteria: b, bacillus subtilis: photosynthetic bacteria: actinomycetes: trichoderma koningii ═ 1:1:4:1:4: 2.
Further limiting, the effective dose of the straw fermentation inoculum is 5 per mill of the total mass of the decomposition product.
The preparation method of the straw fermentation inoculum comprises the following steps: mixing saccharomyces cerevisiae, lactic acid bacteria, bacillus subtilis, photosynthetic bacteria, actinomycetes and trichoderma koningii according to the added mass proportion to obtain the straw fermentation microbial inoculum.
The application of the straw fermentation inoculant in preparation of straw decomposition agents.
The invention provides a method for decomposing straws at low temperature, which comprises the following steps:
(1) activating and preparing a decomposing agent: calculating the amount of the required microbial inoculum according to the total amount of straws, adding 1 kg of brown sugar and 3 kg of urea into 5 times of 25-30 ℃ warm water for reviving for 2-6 hours, and uniformly stirring for later use as a fermentation heat source liquid;
(2) preparing a heat source bag: 3-5kg of rice hulls serving as a matrix, spraying the rice hulls to be wet by using heat source liquid obtained in the step (1) after heating to 42 ℃, wrapping the inner layer by using non-woven fabrics, winding by using a degradable film, and wrapping the outer layer by 2 layers of straw woven fabrics or gunny bags, wherein the volume of the outer layer is 30 multiplied by 30cm 3;
(3) piling: the method comprises the steps of stacking straws in a layered mode, wherein each layer of 1 layer of pig manure or cow manure is 50-60cm high and 5-7cm thick, then spraying 1 layer of the straw fermentation microbial inoculum, controlling the water content to be 60% -65%, stacking the straw fermentation microbial inoculum layer by layer, stacking 4-5 layers of the straw fermentation microbial inoculum in each stack, placing 5 heat source bags in the middle layer in a Z shape, stacking the straw fermentation microbial inoculum for 2-2.5 meters to form a ladder-shaped stack, turning the stack after fermenting for 40-50 days, turning the unfermented straws on the outer layer into the stack, and continuing fermenting for 40 days.
Has the advantages that: the straw fermentation inoculant provided by the invention can complete straw composting fermentation at a low temperature of-20-15 ℃, can complete fermentation in about 2-3 months under the low temperature conditions of spring and autumn, and is simple to operate and low in cost: the rice straw is thoroughly decomposed, and the operation can be carried out in the field without crushing or building a fermentation tank.
Drawings
FIG. 1 shows the fermentation heap with sunken surface and fermented inside.
FIG. 2 shows the enlargement of the subsidence of the surface of the fermentation pile and the further fermentation inside the fermentation pile.
FIG. 3 shows the process of opening the fermentation pile with a forklift to detect the fermentation condition in the pile.
FIG. 4 shows the basic completion of the fermentation of corn stover.
FIG. 5 is the situation after the fermentation of corn stalks is completed.
DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION
Inoculating the saccharomyces cerevisiae strain in a basic fermentation culture medium: peptone 20g, glucose 20g, KH2PO4 1.0g、Mg SO4·7H20.5g of O and 0.5g of Na Cl, and the culture temperature is 30 ℃ and 3 days;
the actinomycete strain is inoculated in a Gao's I culture medium: soluble starch 20.0g, KNO31.0g, K2HPO4 0.5g、Mg SO4·7H20 0.5g、FeSO4·7H20.1g of O and 0.5g of NaCI, the culture temperature is 28 ℃, and the culture time is 3 days;
inoculating the Corning Trichoderma strain in a liquid fermentation culture medium: 5.0g of peptone, 2.5g of yeast extract, 1.0g of glucose and 5.0g of sodium chloride, and the culture temperature is 16 ℃ and 6 days;
inoculating bacillus subtilis in a beef extract peptone culture medium: 3g of beef extract, 5g of peptone and 5g of NaCI, and the culture temperature is 37 ℃ and 3 days;
light and bacteria inoculation in culture medium: NH (NH)4Cl 1.0g、K2HPO4 0.5g、Mg Cl20.2g, Na Cl 2g and 0.1g of yeast extract, wherein the culture temperature is 37 ℃, and the culture time is 3 days;
lactic acid bacteria in culture medium: peptone 10g, beef extract 10g, yeast powder 5g, K2HPO42g of lemon diammonium, 2g of sodium acetate, 5g of glucose, 801mL of Tween and MgSO4·7H2O 0.58g、MnSO4·7H2O0.25 g, culture conditions 30 ℃, 3 days. The inoculum size was 5%, shaking at 28 deg.C in a full temperature shaker, and oscillating at 120 r/min.
The bacteria of the present invention are all purchased commercially.
Example 1.
1. Strain screening and determination of filter paper strip disintegration force by strain screening
Strain screening and culturing: collecting rotten leaves under the forest, rotten straws in the field, rotten cow dung and natural straw compost natural fermentation products as separation samples, and separating filamentous fungi by adopting a straw fishing separation method; bacteria and actinomycetes are separated by an enrichment method. Through long-term screening and low-temperature domestication and breeding, 6 strains are respectively obtained, namely NH1 Saccharomyces cerevisiae Saccharomyces cerevisiae, NH2 lactic acid Bacteria, NH3 Bacillus subtilis, NH4 Photosynthetic Bacteria (Photosynthetic Bacteria) and NH5 actinomycetes (actinomycetes and NH6 Trichoderma koningii).
And respectively inoculating the optimal combined bacteria saccharomyces cerevisiae, lactic acid bacteria, bacillus subtilis, photosynthetic bacteria, actinomycetes and trichoderma koningii obtained by the screening experiment of the composite microbial inoculum into corresponding culture media. And adjusting the inoculation and culture time of each strain to obtain a bacterial liquid at the same time. And (3) carrying out amplification culture on each zymophyte liquid according to the mass ratio of saccharomyces cerevisiae: lactic acid bacteria: b, bacillus subtilis: photosynthetic bacteria: actinomycetes: trichoderma koningii 1:1:4: 1; mixing at a ratio of 4:2 to obtain the compound microbial agent.
Determining cellulose decomposition activity by weight loss method, transferring sterilized filter paper to filter paper disintegration experiment culture medium by aseptic operation, and taking filter paper strip disintegration culture medium without inoculation as control; inoculating 5mL of different microbial agents into 100mL of each treated filter paper strip disintegration culture medium, repeating each group for 3 times, carrying out shaking culture at 38 ℃ for 180r/min, observing and recording the time for the filter paper strips to start disintegration, centrifuging at 7000/g after 10 days, pouring supernatant, washing with a mixed solution of hydrochloric acid and nitric acid to eliminate thalli, centrifuging, washing with clear water, centrifuging, drying at 105 ℃, weighing, and calculating the weight loss rate. The filter paper disintegration test can determine the degradation effect of the strain on the cellulose, and the larger the weight loss rate is, the better the degradation effect is. Test results show that 6 strains have a degradation effect on cellulose, wherein the NH3 strain begins to disintegrate within 20 hours, and the weight loss rate can reach as high as 24.2%.
TABLE 1 disintegration Effect of different bacterial strains on Filter paper strips
2. Effect test of fermentation inoculum for degrading water corn straw
Preparation of the microbial agent: after the single liquid microbial fermentation agent is prepared, saccharomyces cerevisiae, lactic acid bacteria, bacillus subtilis, photosynthetic bacteria, actinomycetes and trichoderma koningii are mixed according to the table 2 to prepare a compound microbial agent (FF) to realize compound bacteriaBiomass and the optimization of the proportion of different strains thereof. The experiment was set at 6-factor (6 strains) 4 levels (4 strains 1, 2, 3, 4). Soaking corn stalk in sterilized distilled water for 24 hr, drying, cutting into 2cm pieces, adding 5g into triangular flask, and adding (NH) into each flask4)2SO40.2g、MgS047H215mL of a 5mmol/L PBS buffer solution (O0.05g, pH 7.0). Inoculating the composite bacteria of different proportions into a triangular flask filled with corn straws according to the inoculation amount of 9 percent, taking the non-inoculated bacteria as a blank Control (CK), uniformly mixing, placing at the constant temperature of 14 ℃ for culture, periodically observing the morphological change of the rice straws, and measuring the degradation rate after 20 days. Washing with sterile steam-fed water to remove thallus and spores, filtering, drying the residue in an oven, weighing the undegraded corn stalks respectively by an electronic balance, and recording the mass as Ml(g) And the mass of the corn straw before fermentation is recorded as M0(g) And then:
TABLE 2 determination of straw degradation effect by different ratio of each strain32(46) Orthogonal table
Example 2 fermentation inoculum application
Adding straws and a mixture of the straws and the livestock manure in a mass ratio of 2:1, and converting the straws and the mixture by solid fermentation to obtain a decomposed organic product. The technical product formula microbial inoculum mainly takes large pile fermentation of more than 50 tons, and the phenomenon that the fermentation cannot be performed due to air drying and complete freezing is prevented. Adding zymocyte according to the proportion of five thousandths of the total mass, adjusting the moisture of the straw to about 65 percent, compacting and fermenting. In winter, hot spot fermentation is needed, namely 100 kg of rice hull powder is fermented firstly to prepare a hot spot bag, and the hot spot bag is placed at the center of a straw fermentation pile to guide fermentation. Turning over after fermenting for about 40-50 days, turning over the outer layer of unfermented straws to the inside, and continuing fermenting for more than 40 days.
1. Application method of zymophyte agent
2. Time: in 2019, 11 and 15 days, in Qing' an county, and in 2020, 10 and 28 days, in Longjiang county, large-scale field tests were performed.
3. Piling: and selecting a convenient place for water taking on the field side. In contrast, the corn straws are used as materials, the amount of the straws in the stack is more than 50 tons, and the amount of organic manure such as pig manure or cow manure is 25 tons. And adding five thousandths of compound bacteria of the total mass of the control pile body on the basis of the control of the treatment pile body. When stacking, the straws are stacked in layers, wherein each layer is 50-60cm high, 1 layer of pig manure or cow manure is 5-7cm thick, then 1 layer of compound bacteria is sprayed, and water is fully sprayed to enable the water content to be 60% -65%, the straws are stacked layer by layer, 4-5 layers can be stacked in each stack, 5 heat source bags are placed in the middle layer in a Z shape, and the straws are stacked for 2-2.5 meters to form a ladder-shaped stack. The compaction is covered by a film to facilitate moisture retention and heat preservation. In winter, hot spot fermentation is needed, namely 100 kg of rice hull powder is fermented firstly and placed in the center of the straw fermentation pile to guide fermentation. Turning over the pile after fermenting for about 40-50 days, turning over the unfermented straws on the outer layer to the inside, and continuing fermenting for more than 40 days. FIG. 1 shows the fermentation heap with sunken surface and fermented inside. FIG. 2 shows the process of opening the fermentation pile with a forklift to detect the fermentation condition in the pile. FIG. 3 shows the process of opening the fermentation pile with a forklift to detect the fermentation condition in the pile. FIG. 4 shows the basic completion of the fermentation of corn stover. FIG. 5 is the situation after the fermentation of corn stalks is completed.
4. Activating and preparing a decomposing agent: calculating the amount of the required microbial inoculum according to the total amount of straws, and then reviving for 2-6 hours by using warm water (the water temperature is 42 ℃, 1 kg of brown sugar and 3 kg of urea are added) of which the amount is 5 times. Stirring uniformly for later use as a fermentation heat source liquid.
5. A heat source bag: the volume of the size is 30 x 30cm3And the core is as follows: crushing corn stalks and rice husks, and spraying heat source liquid at about 42 ℃ for wetting; inner layer: wrapping with non-woven fabric, and winding with degradable film; outer layer: 2 layers are wrapped by straw-woven fabrics or gunny bags.
6. The application effect of the fermentation inoculum is as follows:
(1) change of straw color in the process of decomposition
As a result, as shown in Table 3, the corn stover before treatment was yellow in appearance. At 10d, none of the 3 treated corn stover changed significantly; after 40 days, investigation is carried out, the color of the test straw with the missed bacteria as a reference is not changed, and the color of the test straw inoculated with the composite microbial inoculum is changed from yellow to brown; after 50 days, test straws (the middle part of the stack body) inoculated with the microbial inoculum are further decomposed, and the color of the straws is changed into grey brown; after 60 days, the test straws inoculated with the microbial inoculum are basically decomposed, and the straws are further decomposed and turn into dark brown.
TABLE 3 color change of straw during decomposition
| Treatment of | Before application | 10 days | 25 days | 40 days | 50 days | 60 days |
| CK | Yellow colour | Yellow colour | Yellow colour | Light yellow | Light gray yellow | Light brown |
| Process 1 (Qingan) | Yellow colour | Yellow colour | Yellow colour | Brown colour | Grey brown | Dark brown color |
| Treatment 2 (Sublization) | Yellow colour | Yellow colour | Yellow colour | Brown colour | Grey brown | Dark brown color |
| Treatment 3 (Longjiang) | Yellow colour | Yellow colour | Light yellow | Brown colour | Grey brown | Dark brown color |
(2) Temperature change of the compost during the process of decomposition
The results are shown in table 4, compared with the blank control without straw-degrading bacteria, the temperature of the straw in the test stack is obviously increased after the microbial inoculum is inoculated; treatment 1 increased the temperature the fastest, reaching 52 ℃ on day 10 and 63 ℃ maximum on day 25. The maximum temperature reached in treatment 2 was 61 ℃ and the maximum temperature reached in treatment 3 was 58 ℃.
TABLE 4 Effect of treatments on compost temperature
(2) The decomposing effect of the fermentation inoculum is as follows: as shown in tables 5 and 6, the fermentation process of Qing' an, seiuhuan and Longjiang in 2019 and 2020 takes 50-60 days, the volume of the stack is gradually reduced in the process of the composting reaction, the volume is about initial 2/3 when the composting is finished, and the color of the straw is gradually deepened, so that the straw is obviously weightless and has small volume. The total C/N of the stable or decomposed compost should be less than or equal to 20 by test analysis, the T value of the compost (the ratio of the end point total C/N to the initial C/N) is less than 0.60, and the total carbon content and the total nitrogen content are C/N. The T values of the three treatments are respectively 0.40, 0.43 and 047, and the straw degradation rates are respectively 58.9%, 54.7% and 51.9%. The straw can be applied to the field before sowing after being thoroughly decomposed, and the normal sowing operation is ensured.
TABLE 5 analysis and comparison of carbon and nitrogen test for each treatment
TABLE 6C/N comparison and degradation rates for various treatments
| Item | Initial C/N | End point full C/N | Value of T | The degradation rate% |
| Treatment of 1 (Qingan) pigs | 30 | 11.93 | 0.40 | 58.9 |
| Treatment of 2 (Sublinized) cattle | 30 | 12.88 | 0.43 | 54.7 |
| Processing 3 (Longjiang) cattle | 30 | 14.01 | 0.47 | 51.9 |
| Control 4 | 30 | 30 | -- | -- |
1. Low temperature resistance: finishing the straw composting fermentation at the low temperature of-20-15 ℃. 2. The fermentation time is short: the fermentation can be completed in about 2-3 months under the low temperature condition of spring and autumn. 3. The operation is simple, the cost is low: the rice straw is thoroughly decomposed, and the operation can be carried out in the field without crushing or building a fermentation tank.
Claims (9)
1. The low-temperature-resistant straw fermentation inoculant is characterized by comprising the following components: saccharomyces cerevisiae (Saccharomyces cerevisiae), lactic acid Bacteria (Lactobacillus), Bacillus subtilis (Bacillus subtilis), Photosynthetic Bacteria (Photosyntetic Bacteria), Actinomycetes (Actinomycete), and Trichoderma koningii (Trichoderma koningii).
2. The straw fermentation inoculant according to claim 1, further comprising livestock manure and straw.
3. The straw fermentation inoculum according to claim 2, wherein the mass ratio of straw to animal manure is 2: 1.
4. The straw fermentation inoculant according to claim 2, wherein the straw moisture is adjusted to 65%.
5. The straw fermentation inoculant according to claim 1, wherein the straw fermentation inoculant consists of the following components in percentage by mass: brewing yeast: lactic acid bacteria: b, bacillus subtilis: photosynthetic bacteria: actinomycetes: trichoderma koningii ═ 1:1:4:1:4: 2.
6. The straw fermentation inoculant according to claim 1, wherein the effective dose of the straw fermentation inoculant is 5% o of the total mass of the decomposition product.
7. The preparation method of the straw fermentation inoculum according to any one of claims 1 to 6, which is characterized by comprising the following steps: mixing saccharomyces cerevisiae, lactic acid bacteria, bacillus subtilis, photosynthetic bacteria, actinomycetes and trichoderma koningii according to the added mass proportion to obtain the straw fermentation microbial inoculum.
8. The use of the straw fermentation inoculant according to any one of claims 1-6 in the preparation of a straw decomposition inoculant.
9. A method for decomposing straws at low temperature is characterized by comprising the following steps:
(1) activating and preparing a decomposing agent: calculating the amount of the required microbial inoculum according to the total amount of straws, adding 1 kg of brown sugar and 3 kg of urea into 5 times of 25-30 ℃ warm water for reviving for 2-6 hours, and uniformly stirring for later use as a fermentation heat source liquid;
(2) preparing a heat source bag: 3-5kg rice hull as matrix, spraying heat source liquid obtained in step (1) by heating to 42 deg.C, wrapping the inner layer with non-woven fabric, winding with degradable film, wrapping the outer layer with 2 layers of straw woven fabric or gunny bag, and volume of 30 × 30 × 30cm3；
(3) Piling: stacking the straws in layers, wherein each layer is 50-60cm high, 1 layer of pig manure or cow manure is 5-7cm thick, then spraying 1 layer of the straw fermentation inoculant according to claim 1, controlling the water content to be 60-65%, stacking the straw fermentation inoculant layer by layer, stacking 4-5 layers of straw fermentation inoculant layer by layer, placing 5 heat source bags in the middle layer in a Z shape, stacking the straw fermentation inoculant layer for 2-2.5 meters, stacking the straw fermentation inoculant layer in a trapezoidal shape, turning the straw fermentation inoculant layer after fermenting for 40-50 days, turning the straw fermentation inoculant layer to the inside, and continuing fermenting for 40 days.
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