CN112574928A - Cold region straw decomposition microbial inoculum and preparation method and application thereof - Google Patents
Cold region straw decomposition microbial inoculum and preparation method and application thereof Download PDFInfo
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- CN112574928A CN112574928A CN202110007359.1A CN202110007359A CN112574928A CN 112574928 A CN112574928 A CN 112574928A CN 202110007359 A CN202110007359 A CN 202110007359A CN 112574928 A CN112574928 A CN 112574928A
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Abstract
The invention provides a cold area straw decomposition microbial inoculum, a preparation method and application thereof, and belongs to the technical field of agricultural preparations. The cold-region straw decomposition microbial inoculum comprises a low-temperature flora and a normal-temperature flora; the low-temperature flora comprises lactobacillus plantarum with the cell number content of 20-28%, lactococcus lactis subspecies lactis with the cell number content of 17-25%, lactococcus 14-22%, arthrobacter globiformis with the cell number content of 11-22% and rhodococcus 9-18%; the normal-temperature flora is lignocellulose decomposition flora. The product provided by the invention can start fermentation under a low-temperature condition, promote the decomposition of straws under the low-temperature condition, improve the in-situ decomposition efficiency of the straws in the cultivated land, and solve the difficult problems of dead seedlings and yield reduction caused by severe straw decomposition in summer of the paddy field in cold areas in China.
Description
Technical Field
The invention belongs to the technical field of agricultural preparations, and particularly relates to a cold region straw decomposition microbial inoculum, and a preparation method and application thereof.
Background
Straw is the most productive renewable resource in the world. According to incomplete statistics, the annual straw yield of the world is 850 hundred million tons, and the annual straw yield of crops in China only reaches 8.5 hundred million tons. Lignocellulose in straw is bound together by cellulose, hemicellulose, lignin, ash and the like, which makes cellulose difficult to hydrolyze, and is thus compared to "concrete". The amount of straws in the main production area of the northeast grain is huge, the marginal effect of the yield and the input ratio of the grain in the ploughing area is rapidly reduced, the degraded black soil is urgently needed to be fertilized, and the straw returning decomposition microbial inoculum is urgently needed to improve the straw decomposition efficiency under the low-temperature condition.
In northeast regions, the temperature is 3-6 ℃ per year, a large amount of waste straws are slowly decomposed under the low-temperature condition, and are randomly piled up to occupy cultivated land, thereby not only wasting precious resources, but also harming the ecological environment of rural areas. If a large amount of residual straws in a grain production area are not treated and directly returned to the field or are decomposed and returned to the field by adopting the existing microbial inoculum, the problems of seedling burning and yield reduction caused by severe decomposition of the straws to generate heat and concentrated release of gas in a high temperature period in summer and the problem that the growth of the crops in the current period and the next period is influenced by the incompletely decomposed straws can be caused. Although the average annual temperature in northeast regions is 3-6 ℃, the temperature in spring and summer of each year is rapidly increased in 6-9 months, particularly the lowest temperature in days of 7-8 months is 18-21 ℃, the highest temperature in days is 27-31 ℃, the average monthly temperature is 20-29 ℃, microorganisms decomposing straws in cultivated land are gradually activated along with the temperature increase, and biological heat and gas generated by the microorganisms decomposing the straws violently are released and damage crop roots in a centralized manner, so that root tissues are damaged or loosened, the plants are yellowed to generate seedling burning and death, the seedling burning phenomenon is particularly prominent in paddy fields, and the yield of paddy rice is reduced by 20-40% in severe cases. In order to avoid the reduction of yield, farmers need to discharge water in the paddy field and then return the water in 7-8 months every year, and the operation is repeated for 2-3 times, so that the investment of manpower, material resources and the like is increased.
The present invention relates to a low-temperature straw decomposing bacterium, such as: the invention with the application number of CN201610224388.2 discloses a compound microbial agent for low-temperature fermentation of straws, which comprises Trichoderma viride, Aspergillus niger, Bacillus subtilis, lactococcus lactis, Pediococcus acidilactici and Bacillus circulans. The microbial inoculum can decompose and blacken straws in about 12 days at a low temperature of 4-15 ℃, and the straws are basically decomposed. However, the application range of the microbial inoculum is that the straws are collected to prepare compost, and the microbial inoculum is not suitable for in-situ decomposition and returning of the straws in cultivated land, and farmland experimental results in northern cold regions show that the effect of the CN201610224388.2 on in-situ decomposition of the straws in the cultivated land is not ideal.
The invention with the application number of CN201811581735.2 discloses a low-temperature degradation resistant cellulose composite microbial inoculum, which comprises flavobacterium johnsonii and stenotrophomonas maltophilia, wherein the total effective viable count in the low-temperature degradation resistant cellulose composite microbial inoculum is 3 multiplied by 109The degradation rate of the microbial inoculum is more than 50 percent in 45 days at the temperature of 8-16 ℃. Although the application range of the microbial inoculum is the in-situ decomposition returning microbial inoculum, the application amount of the microbial inoculum is 4.5 kg/mu, the dosage of the low-temperature cellulose decomposition microbial inoculum in the northeast area is 0.5-1 kg/mu, and CN201811581735.2 the dosage of the microbial inoculum is 4.5-9 times of the dosage of the conventional microbial inoculum. In actual use, the microbial inoculum consumes higher manpower and economic cost for transportation, spraying and the like.
Disclosure of Invention
In view of the problems in the background art that the conversion efficiency of straws is low in a cold area or under a low-temperature condition, the decomposition of the conventional microbial inoculum in situ returning is slow, and a series of problems that the growth of crops is affected due to severe root burning, seedling burning, yield reduction and the like caused by straw decomposition occur in a high-temperature straw concentrated decomposition period in summer, the invention provides a cold area straw decomposition microbial inoculum, a preparation method and application thereof. The cold-region straw decomposition microbial inoculum provided by the invention can be quickly fermented under a low-temperature condition, promotes the decomposition and conversion of straws under the low-temperature condition, and avoids the problems of seedling injury and yield reduction caused by concentrated release of biological heat or gas generated by violent straw decomposition in spring end and high-temperature period in summer.
The invention provides a cold-region straw decomposition microbial inoculum, which comprises a low-temperature flora and a normal-temperature flora; the low-temperature flora comprises lactobacillus plantarum with the cell number content of 20-28%, lactococcus lactis subspecies lactis with the cell number content of 17-25%, lactococcus 14-22%, arthrobacter globiformis with the cell number content of 11-22% and rhodococcus 9-18%; the normal-temperature flora is lignocellulose decomposition flora.
Preferably, the low-temperature flora is prepared into freeze-dried powder or is embedded in microcapsules.
Preferably, the freeze-dried powder or the microcapsule carrier comprises a low-temperature bacterium protective agent and a low-temperature bacterium antioxidant; the low-temperature bacterium protective agent comprises one or more of skim milk powder, alpha-glucopyranose, NaCl, glycerol, lactose, polyethylene glycol and trehalose; the low-temperature bacterium antioxidant comprises one or more of vitamin E, tocopherol, cysteine hydrochloride, sodium ascorbate, tert-butyl hydroquinone, ascorbyl palmitate, isoascorbic acid and sodium erythorbate; the mass ratio of the low-temperature bacterium protective agent to the low-temperature bacterium antioxidant is 3-20: 1; the mass of the low-temperature flora thallus is 6-8 times of the sum of the mass of the low-temperature flora protective agent and the mass of the low-temperature flora antioxidant.
Preferably, the microcapsule carrier further comprises a matrix material and a coagulant; what is needed isThe matrix material comprises sodium alginate and/or chitosan; the coagulant comprises CaCl2。
Preferably, the normal-temperature flora comprises 20-27% of aspergillus oryzae, 17-24% of penicillium oxalicum, 14-21% of trichoderma viride, 3-9% of penicillium decumbens, 11-18% of bacillus subtilis, 8-15% of clostridium cellulolyticum, 5-12% of bacillus cereus, 3-5% of saccharomyces cerevisiae and 3-5% of pichia guilliermondii.
Preferably, the normal-temperature flora is mixed with a normal-temperature flora protective agent and a normal-temperature flora antioxidant to prepare normal-temperature flora powder; the normal temperature bacteria protective agent comprises one or more of glycerol, polyethylene glycol, peptone, tween, trehalose, anhydrous sodium acetate, dipotassium hydrogen phosphate, magnesium sulfate and manganese sulfate; the room temperature bacteria antioxidant comprises one or more of shiitake mushroom scrap, white mushroom scrap, Chelidonium majus scrap, waste tea powder, Salvia miltiorrhiza powder, licorice scrap, clove scrap, monkshood powder, gallic acid powder and superoxide dismutase SOD; the mass ratio of the room-temperature bacterium protective agent to the room-temperature bacterium antioxidant is 1 (1-10); the mass of the normal-temperature flora thallus is 6-7 times of the sum of the mass of the normal-temperature flora protective agent and the mass of the normal-temperature flora antioxidant.
Preferably, the ratio of the cell number of the low-temperature flora to the cell number of the normal-temperature flora is 1 (3-6).
The invention provides a preparation method of the cold-region straw decomposition microbial inoculum, which comprises the step of flora culture, wherein the flora culture comprises low-temperature flora culture and normal-temperature flora culture; the culture medium for low-temperature flora culture comprises 5-25 g/L of carbon source and K2HPO40.8～1.2g/L，KH2PO40.8-1.2 g/L, 2-10 g/L peptone, 2-10 g/L beef extract, 1-5 g/L, MgSO yeast powder4·7H20.08-0.12 g/L of O, 1.8-2.2 g/L of sodium acetate and 1.8-2.2 g/L, MnSO of diammonium hydrogen citrate4·H20.04-0.06 g/L of O and 800.8-1.2 mL/L of Tween; the carbon source comprises glucose and/or sucrose; the culture medium for culturing the normal-temperature flora comprises (NH)4)2SO41.2～1.6g/L，MgSO4·7H2O 0.2～0.4g/L，KH2PO41.8～2.2g/L，CaCO31.8-2.2 g/L, peptone 2.2-2.8 g/L, FeSO4·7H2O 4～6mg/L，MnSO41.2～2mg/L，ZnCl21.4～2mg/L，CoCl21.4-2 mg/L and 6-10 g/L of straw.
The invention provides an application of the cold-region straw decomposition microbial inoculum in straw decomposition.
Preferably, the application comprises: the cold-region straw decomposition microbial inoculum is applied to straw in-situ returning, and the application amount of the cold-region straw decomposition microbial inoculum is 0.5-1.2 kg/mu.
Has the advantages that: the invention provides a cold region straw decomposition microbial inoculum and a preparation method and application thereof. The low-temperature decay microbial inoculum comprises 1 low-temperature flora and 1 normal-temperature lignocellulose decomposition flora, wherein the low-temperature flora can grow and reproduce at the temperature of 1-4 ℃, and the optimal growth temperature is 10-15 ℃. The biological heat energy generated in the growth, reproduction and metabolism processes of the low-temperature flora provides a proper temperature environment for the growth of the normal-temperature flora, and the normal-temperature flora, namely the lignocellulose decomposition flora, metabolizes and converts lignocellulose in the growth and reproduction processes.
In a preferred scheme, freeze-dried powder or a microcapsule carrier is adopted to embed the low-temperature flora, so that the damage of room temperature or high temperature to lipid and protein molecules in cell membranes of the low-temperature flora in the storage process is reduced, the activity of thalli is protected, the survival rate of the microbial inoculum of the low-temperature flora at the room temperature is improved, and the activity of the microbial inoculum is kept. The microcapsule treatment can keep the activity of the low-temperature flora for a long time, so that the service life of the low-temperature flora produced in the same batch is matched with that of the normal-temperature flora. The product prepared by the invention can keep the survival rate of the bacterial strain, the diversity of the flora composition, the health state of the bacterial strain and the performance of decomposing straws for a long time.
The cold-region straw decomposition microbial inoculum is applied to straw decomposition (particularly paddy field in-situ decomposition returning and fertilizing farmlands), straw decomposition reaction is started in the farmlands by the microbial inoculum in 4-5 months in the cold regions, gas and heat generated by straw decomposition are released at any time, and damage to seedling roots and plants by the gas and heat generated by concentrated and violent decomposition of the straws in 7-9 months can be effectively relieved or reduced.
The technology of the straw decomposition microbial inoculum and the like in the cold area can decompose the straws into humus in situ without moving the straws out of the cultivated land and return the humus to soil, effectively promotes the circulation of organic wastes for farming and pasturing, avoids the links of transporting the organic fertilizers back to the cultivated land and the like after the straws are transported out of the cultivated land and are intensively piled up to produce the organic fertilizers, greatly reduces the cost of manpower, energy power and the like consumed by transportation, piling up and transportation back to the farmland in the process of preparing the organic fertilizers by the straws, provides technical support for improving the problems of deterioration of the cultivated land in the cold area, comprehensive utilization of the straws and the like, and is favorable for promoting the sustainable development of.
Biological preservation information description
The microorganisms with the deposit numbers in the embodiments of the present invention are all available from the corresponding depository. The preservation institutions and addresses involved are as follows: china general microbiological culture Collection center (CGMCC), with the address of No. 3 Xilu No. 1 Beijing, Chaoyang district; china agricultural microbial strain preservation management center (ACCC) with the address of No. 12 southern street of Guancun in Haita district, Beijing; china center for the preservation and management of industrial microbial strains (CICC) at the address of No. 6 Hospital No. 24 of Zhonglu Jiuxiangqiao of Chaoyang district, Beijing.
Drawings
FIG. 1 is a graph of weight loss analysis of rice straw in an experiment described in example 3 of the present invention;
FIG. 2 is a graph showing the change in the humic substance content of soil in the experiment described in example 4 of the present invention;
FIG. 3 is a graph of organic matter change in cultivated land according to the experiment described in example 5 of the present invention;
FIG. 4 is a graph showing the variation of cellulase content in the experiment described in example 6 of the present invention;
FIG. 5 is a graph showing the activity of endo-cellulase in the experiment described in example 7 of the present invention;
FIG. 6 is a diagram showing the residual straw amount in the experiment of example 7 of the present invention;
FIG. 7 is a graph showing the survival rate of the 4 low temperature flora treated according to example 8 of the present invention.
Detailed Description
The invention provides a cold-region straw decomposing microbial inoculum which comprises a low-temperature flora and a normal-temperature flora.
The low-temperature flora provided by the invention comprises lactobacillus plantarum (Lactobacillus plantarum), Lactococcus lactis subsp. The cell number content of the lactobacillus plantarum is 20-28%, preferably 24-26%, and more preferably 25%; the cell number content of lactococcus lactis subspecies lactis is 17-25%, preferably 21-23%, and more preferably 22%; the content of the lactococcus cell number is 14-22%, preferably 18-20%, and more preferably 19%; the cell number content of the arthrobacter globiformis is 11-22%, preferably 18-20%, and more preferably 19%; the content of the number of the cells of the rhodococcus is 9-18%, preferably 14-16%, and more preferably 15%. The present invention is not limited to the above-mentioned species, and any species can be obtained from a commercially available source. In a preferred embodiment of the invention, the lactobacillus plantarum has a preservation number of CGMCC-1.12974; the preservation number of the lactococcus lactis subspecies lactis is as follows: CGMCC-1.2030; the lactococcus has the preservation number: CGMCC-1.15072; the preservation number of the arthrobacter globiformis is CICC-23557; the preservation number of the rhodococcus is CICC-20603.
The source and production method of the low temperature flora are not particularly limited in the present invention, and the low temperature flora may be obtained by a conventional method in the art. In an alternative embodiment of the invention: and respectively purchasing independent microbial agents of the bacteria, and mixing according to the cell ratio to obtain the low-temperature microbial community. In an alternative aspect of the invention: and (3) culturing the strains in an improved low-temperature bacterium culture medium at low temperature to obtain the low-temperature bacterium group. According to the invention, 5 kinds of bacteria are preferably mixed according to a preferable proportion and then subjected to long-term low-temperature domestication treatment, wherein the domestication treatment refers to: culturing mixed flora of each bacterium from 20 ℃, periodically detecting the biomass OD of the microorganism, after the biomass OD of the flora is increased to 1.6 and the required time is stable and consistent, reducing the culture temperature by 3 ℃, continuing culturing until the temperature is reduced to 4 ℃, detecting the biomass OD of the microorganism, when the OD is increased to 1.6 and the required time is stable and consistent, obtaining low-temperature flora, and culturing the low-temperature flora at 4 ℃. The temperature of the northern area is about 5-22 ℃ in 4-5 months, and the cultivation at 4 ℃ is beneficial to keeping the growth and reproduction vitality of the low-temperature flora in the low-temperature environment. The culture time of the low-temperature bacteria is preferably 3-15 d, and more preferably 6-8 d.
In the low-temperature flora culture of the present invention, the culture medium for low-temperature flora culture comprises: carbon source 5-25 g/L, K2HPO40.8～1.2g/L，KH2PO40.8-1.2 g/L, 2-10 g/L peptone, 2-10 g/L beef extract, 1-5 g/L, MgSO yeast powder4·7H20.08-0.12 g/L of O, 1.8-2.2 g/L of sodium acetate and 1.8-2.2 g/L, MnSO of diammonium hydrogen citrate4·H2O0.04-0.06 g/L, Tween 800.8.8-1.2 mL/L; preferably, the method comprises the following steps: carbon source 10-20 g/L, K2HPO41.0 g/L，KH2PO41.0 g/L, 4-8 g/L peptone, 4-8 g/L beef extract and 2-4 g/L, MgSO g yeast powder4·7H20.1g/L of O, 2g/L of sodium acetate and 2g/L, MnSO g of diammonium hydrogen citrate4·H2O0.05 g/L, Tween 801.0 mL/L. The carbon source preferably comprises glucose and/or sucrose. The sources of the above components are not particularly limited in the present invention, and any products conventionally commercially available in the art may be used. After the low-temperature flora is cultured, preferably obtaining low-temperature thalli in a centrifugal mode; the rotation speed of the centrifugation is preferably 4000 rpm; the time for centrifugation is preferably 5 min.
The low-temperature bacteria is obtained after the low-temperature flora culture, and preferably, the low-temperature bacteria is mixed with a low-temperature bacteria protective material to obtain a low-temperature bacteria mixture. In the invention, the low-temperature bacterium protective material comprises a low-temperature bacterium protective agent and a low-temperature bacterium antioxidant. The low-temperature bacterium protective agent preferably comprises one or more of skim milk powder, alpha-glucopyranose, NaCl, glycerol, lactose, polyethylene glycol and trehalose, and more preferably the skim milk powder or the glycerol; the cryopreserved antioxidant preferably comprises one or more of vitamin E, tocopherol, cysteine hydrochloride, sodium ascorbate, tertbutylhydroquinone, ascorbyl palmitate, erythorbic acid and sodium erythorbate, more preferably vitamin E or sodium ascorbate. The mass ratio of the low-temperature bacterium protective agent to the low-temperature bacterium antioxidant is preferably 3-20: 1, more preferably 4-10: 1, and more preferably 5: 1. The mass of the low-temperature thallus is preferably 6-8 times, and more preferably 7 times of the sum of the mass of the low-temperature thallus protective agent and the mass of the low-temperature thallus antioxidant.
After the low-temperature bacterium mixture is obtained, the low-temperature bacterium mixture is preferably prepared into microcapsules or freeze-dried powder. The preparation method of the microcapsule or the freeze-dried powder is not particularly limited, and the microcapsule or the freeze-dried powder can be prepared by conventional methods in the field.
The normal temperature flora is lignocellulose decomposing flora, and preferably comprises Aspergillus oryzae (Aspergillus oryzae), Penicillium oxalicum (Penicillium oxalicum), Trichoderma viride (Trichoderma viride), Penicillium decumbens (Penicillium decumbens), Bacillus subtilis (Bacillus subtilis), Clostridium cellulolyticum (Clostridium cellulolyticum), Bacillus cereus (Bacillus cereus), Saccharomyces cerevisiae (Saccharomyces cerevisiae), and Pichia guilliermondii (Pichia guilliermondii). The content of the number of the cells of the Aspergillus oryzae is preferably 20-27%, more preferably 21-24%, and more preferably 22%; the content of the number of the penicillium oxalicum cells is preferably 17-24%, more preferably 18-21%, and more preferably 19%; the content of the number of the cells of the trichoderma viride is preferably 14-21%, more preferably 15-18%, and more preferably 16%; the content of the number of the cells of the penicillium decumbens is preferably 3-9%, more preferably 4-7%, and more preferably 5%; the content of the number of the cells of the bacillus subtilis is preferably 11-18%, more preferably 12-15%, and more preferably 13%; the cell number content of the clostridium cellulolyticum is preferably 8-15%, more preferably 9-12%, and more preferably 10%; the content of the number of the cells of the bacillus cereus is preferably 5-12%, more preferably 6-8%, and more preferably 7%; the cell number content of the saccharomyces cerevisiae is preferably 3-5%, and more preferably 4%; the cell number content of the pichia guilliermondii is preferably 3-5%, and more preferably 4%. The present invention is not limited to the above-mentioned species, and any species can be obtained from a commercially available source. In a preferred embodiment of the invention, the Aspergillus oryzae has a accession number of CGMCC-3.7203; the preservation number of the penicillium oxalicum is as follows: CGMCC-3.15651; the preservation number of the trichoderma viride is as follows: CGMCC-3.3744; the preservation number of the penicillium decumbens is CGMCC-3.3195; the preservation number of the bacillus subtilis is CGMCC-1.15792; the preservation number of the clostridium cellulolyticum is ACCC-00528; the preservation number of the bacillus cereus is as follows: CICC-10042; the preservation number of the saccharomyces cerevisiae is CICC-1012; the Pichia guilliermondii has a deposit number of CICC-1951.
The method for preparing the normal temperature flora is not particularly limited, and the normal temperature flora obtained by the conventional method in the field can play corresponding roles. In an alternative embodiment of the invention: respectively purchasing independent microbial agents of the above bacteria, and mixing according to the ratio of cell numbers to obtain the normal temperature microbial community. In an alternative aspect of the invention: the normal temperature flora is obtained by a normal temperature flora culture mode.
In the culture of normal temperature flora according to the present invention, the culture medium for normal temperature flora culture includes: (NH)4)2SO41.2～1.6g/L，MgSO4·7H2O 0.2～0.4g/L，KH2PO41.8～2.2g/L，CaCO31.8-2.2 g/L, peptone 2.2-2.8 g/L, FeSO4·7H2O 4～6mg/L，MnSO41.2～2mg/L，ZnCl21.4～2mg/L，CoCl21.4-2 mg/L and 6-10 g/L of straw; preferably, the method comprises the following steps: (NH)4)2SO41.4g/L，MgSO4·7H2O 0.3g/L，KH2PO42.0 g/L，CaCO32.0 g/L, peptone 2.5g/L, FeSO4·7H2O 5.0mg/L，MnSO41.6 mg/L，ZnCl21.7 mg/L，CoCl21.7 mg/L and 8g/L straw. In the present invention, the pH of the culture medium for culturing normal temperature flora is preferably adjusted to 7.2. The sources of the above components are not particularly limited in the present invention, and any products conventionally commercially available in the art may be used. In the invention, the temperature for culturing the normal-temperature flora is preferably 20-30 ℃, more preferably 21-29 ℃, and more preferably 25-28 ℃; culturing the normal temperature floraThe time of (a) is preferably 10 to 20 days, and more preferably 14 to 15 days. After the normal-temperature flora is cultured, normal-temperature thalli are obtained preferably in a centrifugal mode; the rotation speed of the centrifugation is preferably 4000 rpm; the time for centrifugation is preferably 5 min.
The normal-temperature bacteria is obtained after the normal-temperature flora culture, and preferably, the normal-temperature bacteria is mixed with a normal-temperature bacteria protective material to obtain a normal-temperature bacteria mixture. In the invention, the room temperature bacterium protective material comprises a room temperature bacterium protective agent and a room temperature bacterium antioxidant. The normal temperature bacteria protective agent preferably comprises one or more of glycerol, polyethylene glycol, tween, trehalose, peptone, anhydrous sodium acetate, dipotassium hydrogen phosphate, magnesium sulfate and manganese sulfate, and more preferably glycerol; the room temperature fungus antioxidant preferably comprises one or more of shiitake mushroom scraps, white mushroom scraps, chelidonium scraps, waste tea powder, salvia miltiorrhiza powder, liquorice scraps, clove scraps, aconite root powder, gallic acid and superoxide dismutase SOD, and more preferably shiitake mushroom scraps; the mass ratio of the room temperature bacterium protective agent to the room temperature bacterium antioxidant is preferably 1 (1-10), more preferably 1 (2-5), and more preferably 1: 3; the mass of the normal-temperature bacteria is preferably 6-7 times of the sum of the mass of the normal-temperature bacteria protective agent and the mass of the normal-temperature bacteria antioxidant.
After the normal-temperature bacterium mixture is obtained, the normal-temperature bacterium mixture is preferably subjected to vacuum spray drying to obtain normal-temperature bacterium powder. In the invention, the temperature of the air inlet for spray drying is preferably 80-105 ℃, and more preferably 90 ℃.
In the cold-region straw decomposition microbial inoculum, the content ratio of the cell number of the low-temperature flora to the cell number of the normal-temperature flora is preferably 1 (3-6), and more preferably 1: 4. When the low-temperature flora is prepared into microcapsules containing the low-temperature flora protection material, and the normal-temperature flora is prepared into normal-temperature bacterial powder containing the normal-temperature bacterial protection material, the mass ratio of the microcapsules to the normal-temperature bacterial powder is preferably 1-2: 4-8.
The invention provides a preparation method of the cold region straw decomposition microbial inoculum, which comprises the following steps: respectively preparing low-temperature flora and normal-temperature flora to obtain the straw decomposition microbial inoculum for the cold area. In the present invention, the methods for preparing the low-temperature flora and the normal-temperature flora are as described above.
The invention provides an application of the cold-region straw decomposition microbial inoculum in straw decomposition. Preferably, the application comprises: the cold-region straw decomposition microbial inoculum is used for returning the straws to the field in situ. In the present invention, the in-situ returning is preferably a paddy field in-situ returning. Before the application, the cold-region straw decomposition microbial inoculum is preferably pre-cultured. The pre-culture medium preferably contains 0.08-0.14 kg/L of urea and 0.04-0.08 kg/L of brown sugar (or white sugar); the pre-culture time is preferably 1-2 d. When the straw decomposition bactericide is used, the cold area straw decomposition bactericide is preferably sprayed on the surface of the straw, and then the straw is buried in the cultivated land; the depth of the turning and burying is preferably 10-20 cm, and more preferably 15-18 cm; the water content of the soil is preferably 55-65%. In the invention, the application amount of the cold-region straw decomposition microbial inoculum is preferably 0.5-1.2 kg/mu, and more preferably 1 kg/mu. After the cold area straw decomposition microbial inoculum is applied, base fertilizer does not need to be applied, so that the cost for applying the base fertilizer can be saved.
The cold area mainly refers to a cold area with the annual average temperature of 4-6 ℃ in China. The natural temperature of the cold-region straw decomposition microbial inoculum suitable for degrading the straws is preferably 4-32 ℃, more preferably 10-30 ℃ and most preferably 15-28 ℃. The straw decomposition capability of the cold-region straw decomposition microbial inoculum is weakened at an excessively high or excessively low temperature. The straw decomposition microbial inoculum provided by the invention is suitable for cold regions in the north of China, and has certain universality in the central and south of China. The application of the cold-region straw decomposition microbial inoculum for returning the straws to the field reduces the application amount of the original fertilizer by 20-30% compared with the application of the straws to the field without the treatment of returning the straws to the field.
In a more specific implementation mode, the cold-region straw decomposition microbial inoculum provided by the invention is applied to 600 kg/mu in-situ decomposition and returning to the field in the areas of Ningjiang county, Tailai, Heilongjiang province and Sanjiang province, in which the total amount of straws is 500-. Because the temperature is lower in 3-5 months in the north of China, particularly the lowest temperature is 3-11 ℃ in 4-5 months, the highest temperature in days is 15-22 ℃, and the average temperature is 9-21 ℃. And in the temperature range of growth and reproduction of the cold-region straw decomposition microbial inoculum at 9-21 ℃, the microbial inoculum can start field straw decomposition reaction in 4-5 months, the straw is decomposed in 4-5 months to generate gas and heat which are released at any time, and the damage of the heat and the gas generated by intensively decomposing the straw in 7-9 months to seedling roots and plants is relieved or reduced. The previous results show that: the phenomenon of seedling death and seedling burning does not occur in the experimental field where the microbial inoculum is decomposed in situ and returned to the field, and the crop yield is increased by a small margin.
The present invention provides a straw decomposition microbial inoculum for cold regions, and a preparation method and application thereof are described in detail below with reference to examples. The 4 sources of commercially available straw-decomposing bacteria referred to in the control experiments in the examples are as follows: CK 1: beijing Deruifeng agriculture science and technology, Inc. (Tuwei No.); CK 2: heilongjiang grass warming-harvest ecological technology limited (micro 18 straw decomposition agent); CK 3: heilongjiang province Dafeng science and technology development, Inc. (straw manure ferment: ferment No. 3); CK 4: xintai agricultural science and technology development company Limited (Xintai agricultural three-in-one straw decomposition nutrient). It should be noted that the following examples are only illustrative of the technical idea of the present invention and they should not be construed as limiting the scope of the present invention.
Example 1
A cold region straw decomposition microbial inoculum comprises the following components: low-temperature flora microcapsule and normal-temperature bacteria powder of lignocellulose decomposition flora.
(1) Preparing a low-temperature flora microcapsule:
preparing an improved low-temperature bacterium culture medium: preparing to obtain the glucose-containing material with the concentration of 15g/L, K2HPO41.0 g/L，KH2PO41.0 g/L, 6g/L peptone, 6g/L beef extract and 3g/L, MgSO yeast powder4·7H2O0.1g/L, sodium acetate 2g/L, diammonium hydrogen citrate 2g/L, MnSO4·H2O0.05 g/L, Tween 801mL/L modified cryobacteria culture medium. The mixture is subpackaged by 500mL blue cap bottles, and each bottle contains 350 mL. Placing in a refrigerator at 4 ℃ for later use.
② culturing flora: lactobacillus plantarum, lactococcus lactis subspecies lactis, lactococcus lactis, Arthrobacter globiformis and Rhodococcus erythropolis were inoculated into the modified cryobacterium medium. Culturing at 4 ℃ for 8d, and determining the percentage content of the cell number of each strain as follows: 25% of lactobacillus plantarum, 22% of lactococcus lactis subspecies lactis, 19% of lactococcus lactis, 19% of arthrobacter globiformis and 15% of rhodococcus lactis. Centrifuging at 4000rpm for 5min to obtain low temperature flora thallus.
Adding a protective material into the low-temperature flora thallus to obtain a low-temperature flora mixture: the mass ratio of the low-temperature bacterium protective agent, the low-temperature bacterium antioxidant and the low-temperature bacterium thallus is 5:1: 42.
Preparing microcapsules: uniformly mixing 100g of the cryophyte mixture with 50mL of sodium alginate solution with the mass volume concentration (g/mL) of 2%, and dripping 50mL of CaCl with the mass volume concentration (g/mL) of 2.5%2Stirring to obtain gel beads, and solidifying for 45 min. Adding into 50mL chitosan solution with mass volume concentration (g/mL) of 0.3%, stirring and coating for 45min to obtain microcapsule.
(2) Preparation of a bacterial powder preparation of normal-temperature lignocellulose decomposition flora:
preparing a straw decomposition culture medium: is prepared to obtain a catalyst containing (NH)4)2SO41.4 g/L，MgSO4·7H2O0.3g/L，KH2PO42.0 g/L，CaCO32.0 g/L, peptone 2.5g/L, FeSO4·7H2O 5.0mg/L，MnSO41.6 mg/L，ZnCl21.7 mg/L，CoCl21.7 mg/L straw decomposition medium and 8g/L straw decomposition medium. The pH was adjusted to 7.2. The mixture was dispensed into 500mL triangular flasks, and 300mL each was dispensed. Standing at room temperature of 20-25 ℃ for later use.
Culturing normal-temperature lignocellulose decomposition flora: inoculating Aspergillus oryzae, Penicillium oxalicum, Trichoderma viride, Penicillium decumbens, Bacillus subtilis, Clostridium cellulolyticum, Bacillus cereus, Saccharomyces cerevisiae, and Pichia guilliermondii into the straw decomposition culture medium. Culturing for 15d at 20-25 ℃, and determining the percentage content of the cell number of each strain as follows: aspergillus oryzae 22%, Penicillium oxalicum 19%, Trichoderma viride 16%, Penicillium decumbens 5%, Bacillus subtilis 13%, Clostridium cellulolyticum 10%, Bacillus cereus 7%, Saccharomyces cerevisiae 4% and Pichia guilliermondii 4%. Centrifuging at 4000rpm for 5min to obtain normal temperature flora thallus.
Adding protective materials into the normal-temperature flora thallus: glycerol is used as a protective agent, shiitake fragments are used as an antioxidant, and the mass ratio of the protective agent for normal-temperature bacteria, the antioxidant for normal-temperature bacteria and the normal-temperature flora thallus is 1:3: 28.
Preparing bacteria powder: and (3) carrying out low-temperature spray drying on the normal-temperature flora thallus containing the protective material to obtain the bacterial powder. The temperature of an air inlet is 90 ℃, and normal-temperature lignocellulose decomposition bacteria powder is obtained.
(3) Respectively preparing the low-temperature flora microcapsule and the bacterial powder of the normal-temperature lignocellulose decomposition flora, and using the microcapsules and the bacterial powder in combination. When the composite material is used, the mass ratio of the low-temperature flora microcapsule to the bacterial powder of the normal-temperature lignocellulose decomposition flora is 2: 1.
example 2
A cold region straw decomposition microbial inoculum comprises the following components: freeze-dried powder of low-temperature flora and bacteria powder of normal-temperature lignocellulose decomposition flora.
(1) Preparing low-temperature flora freeze-dried powder:
preparing an improved low-temperature bacterium culture medium: preparing to obtain the glucose-containing material with the concentration of 15g/L, K2HPO41.0 g/L，KH2PO41.0 g/L, 6g/L peptone, 6g/L beef extract and 3g/L, MgSO yeast powder4·7H2O0.1g/L, sodium acetate 2g/L, diammonium hydrogen citrate 2g/L, MnSO4·H2O0.05 g/L, Tween 801mL/L modified cryobacteria culture medium. The mixture is subpackaged by 500mL blue cap bottles, and each bottle contains 350 mL. Placing in a refrigerator at 4 ℃ for later use.
② culturing flora: lactobacillus plantarum, lactococcus lactis subspecies lactis, lactococcus lactis, Arthrobacter globiformis and Rhodococcus erythropolis were inoculated into the modified cryobacterium medium. Culturing at 4 ℃ for 8d, and determining the percentage content of the cell number of each strain as follows: 25% of lactobacillus plantarum, 22% of lactococcus lactis subspecies lactis, 19% of lactococcus lactis, 19% of arthrobacter globiformis and 15% of rhodococcus lactis. Centrifuging at 4000rpm for 5min to obtain low temperature flora thallus.
③ adding protective materials into the low-temperature flora thallus: glycerol is used as a protective agent, sodium ascorbate is used as an antioxidant, and the mass ratio of the low-temperature bacterium protective agent to the low-temperature bacterium antioxidant to the low-temperature bacterium thallus is 5:1: 42.
Fourthly, preparing low-temperature flora freeze-dried powder: adding the thallus of the protective material, precooling for 12 hours at-40 ℃, and freeze-drying for 16 hours by a freeze-drying machine to obtain the low-temperature flora freeze-dried powder.
(2) Preparation of bacterial powder preparation for decomposing bacteria flora with lignocellulose at normal temperature. (same as example 1)
(3) Respectively preparing the low-temperature flora freeze-dried powder and the normal-temperature lignocellulose decomposition flora powder, and matching. When the composition is used, the mass ratio of the low-temperature flora freeze-dried powder to the bacteria powder of the normal-temperature lignocellulose decomposition flora is 1: 4.
example 3
The microbial inoculum prepared in example 1 was used as an experimental group, and inoculated with 4 commercially available straw decomposition microbial inoculants CK1, CK2, CK3 and CK4 simultaneously on a culture medium using rice straws as a sole carbon source, fermented at 28 ℃ for 15 days, and the weight reduction capacity of the rice straws was measured. The weight loss analysis of rice straw under different treatments is shown in FIG. 1. As can be seen from fig. 1: the weight loss of the straws treated by the microbial inoculum, CK1, CK2, CK3 and CK4 is respectively 0.72g, 0.67g, 0.63g, 0.58g and 0.54g, and the weight loss rate is respectively 44.2%, 41.10%, 38.65%, 35.58% and 33.12%. The straw conversion capability of the microbial inoculum under the same conditions is higher than that of 4 commercially available microbial inocula used in experiments.
Example 4
The microbial inoculum prepared in example 1 is used as an experimental group, and indoor simulation field experiments are carried out on the microbial inoculum and 4 commercially available straw decomposition microbial inocula CK1, CK2, CK3 and CK 4: 10 jin of soil, 25g of straws, 6L of tap water and 1g of urea are placed in a square box with the specification of 43 x 33 x 15cm, the straws are flatly paved with 2cm thick soil to avoid floating, 50mL of the microbial inoculum, CK1, CK2, CK3 and CK4 are uniformly sprayed into the square box respectively, the square box is placed in a temperature-controlled greenhouse with the temperature of 10-20 ℃ from 1 month 1 in 2020, and tap water with the same quality is sprayed every 2 weeks to increase the water content. The soil humus content is measured on the experimental day, 6 months and 9 months respectively, and the conversion capacity of each microbial inoculum to straws is analyzed within 9 months under the condition that the microbial inoculum is not interfered by the environment, weather and other external factors. The change conditions of the soil humus content in 6 months and 9 months after straw decomposition under different treatments are shown in figure 2: the humus of the microbial inoculum, CK1, CK2, CK3 and CK4 are respectively 29.26g/kg, 27.33g/kg, 22.23g/kg, 19.32g/kg and 19.56g/kg in 6 months, and are respectively 41.56g/kg, 38.90g/kg, 36.49g/kg, 36.36g/kg and 36.33g/kg in 9 months, and compared with 4 commercially available microbial inocula, the microbial inoculum has stronger capacity of generating humus by converting straws into straws.
Example 5
The microbial inoculum prepared in the example 2 is used as an experimental group, and the microbial inoculum and 4 commercially available straw decomposition microbial inocula CK1, CK2, CK3 and CK4 are respectively subjected to a rice field decomposition straw experiment in the Ming-Jiang county hardworking village (belonging to the first accumulated temperature with annual activity accumulated temperature 2600-2The square pond of one mu, two rice field ponds establish the isolation that the earth wall is about 20cm high, 20cm wide between the pond, and every rice field pond interval one non-experimental rice field pond, pond and pond earth wall interval do not carry out the interchange of water and soil, set up 10 mu land altogether, every is handled 2 repeatedly. The total returning-to-field straw amount is 500kg straw/667 m2The straws are crushed to about 7cm, each microbial inoculum is uniformly sprayed on the straws and the cultivated land in the beginning of 4 months according to the optimal use amount and the use method specified by the respective specification, and the straws are tiled, uniform and free from stacking; burying the straws in a 15-18 cm deep farmland through deep ploughing and turning, reducing the total amount of the original fertilizer by 30%, and respectively measuring the change of organic matters in the farmland before returning to the field and after returning to the field for 2 months, 3 months and 5 months.
Under different treatments, the change of the organic matter content of the soil in situ returning for 2 months, 3 months and 5 months is shown in figure 3. As can be seen from fig. 3: the organic matter content in the cultivated land sprayed by the microbial inoculum is higher than that in 4 groups of the control cultivated land. The average organic matter content of the cultivated land before the microbial inoculum is returned to the field is 29.16g/kg, the average organic matter content of the cultivated land after 5 experimental plots are returned to the field for 2 months, 3 months and 5 months is 32.68g/kg, 30.64g/kg and 29.49g/kg respectively, and the organic matters of the cultivated land after the microbial inoculum is applied to the field for 2 months, 3 months and 5 months are 34.70g/kg, 32.50g/kg and 31.51g/kg respectively and are obviously higher than the average value. The comparison of the organic matter content of cultivated land in combination with the graph of FIG. 3 also shows that the straw conversion capability of the microbial inoculum is stronger.
Example 6
The microbial inoculum prepared in the example 2 is used as an experimental group, the microbial inoculum and 4 commercially available straw decomposition microbial inocula CK1, CK2, CK3 and CK4 are respectively applied to straw in-situ returning in the Ministry of Ministry, Ningjiang and Severe village in Tai county, and the cellulase activity in the soil is measured 2 months and 5 months after the straw is returned to the field. The cellulase is a general term of a complex enzyme system capable of converting lignocellulose into glucose, the activity of the cellulase can objectively reflect the strength of the capability of converting lignocellulose, and the cellulase is an important evaluation index of the capability of a microbial inoculum for converting lignocellulose.
The change of the cellulase content in situ returned to the field for 2 months and 5 months under different treatments is shown in figure 4. As can be seen from fig. 4: cellulase in cultivated land sprayed with 5 groups of microbial inoculum is respectively 1.63mg/g, 1.49mg/g, 1.55mg/g, 1.39mg/g and 1.35mg/g in month 2, cellulase is respectively 1.53mg/g, 1.37mg/g, 1.43mg/g, 1.38mg/g and 1.37mg/g in month 5, average values of 4 control bacteria in month 2 and month 5 are respectively 1.49mg/g and 1.41mg/g, cultivated land treated by the microbial inoculum is respectively 1.64mg/g and 1.53mg/g in month 2 and month 5, and the average value is obviously higher than that of the 4 control bacteria, which indicates that the microbial inoculum can decompose straws in low-temperature cold areas of the Heilongjiang.
Example 7
Preparing an improved low-temperature bacterium culture medium: prepared to obtain the product containing 25g/L of sucrose, K2HPO41.0 g/L，KH2PO41.0 g/L, 6g/L peptone, 6g/L beef extract and 3g/L, MgSO yeast powder4·7H20.1g/L of O, 2g/L of sodium acetate and 2g/L, MnSO g of diammonium hydrogen citrate4·H2O0.05 g/L, Tween 801mL/L modified cryobacteria culture medium. The mixture is subpackaged by 500mL blue cap bottles, and each bottle contains 350 mL. Placing in a refrigerator at 4 ℃ for later use. Preparing a straw decomposition medium (L): (NH)4)2SO42.0 g，MgSO4·7H2O 0.3 g，KH2PO42.0 g，CaCO32.0 g, peptone 2.5g, FeSO4·7H2O 5.0mg，MnSO41.6 mg，ZnCl21.7 mg，CoCl21.7mg, pH is adjusted to be neutral, and straw is 10 g/L. The resulting culture medium was sterilized by autoclaving with 500mL flasks, each containing 300 mL. Transferring 20mL of normal temperature lignocellulose decomposition bacteria group and 6mL of low temperature bacteria group cultured for 7d into fresh culture medium to obtain the strainAn agent; inoculating 20mL of normal-temperature lignocellulose decomposition bacteria group and 6mL of sterile fresh culture medium to serve as normal-temperature bacteria; inoculating 20mL of low-temperature bacteria and 6mL of sterile fresh culture medium, treating 3 bottles of low-temperature bacteria, performing static culture at 18-20 ℃, sampling at 1 st, 3 th, 5 th, 8 th and 14 th days respectively, measuring the indexes of endocellulase activity, weight reduction of rice straws and the like, and verifying the conversion difference of the microbial inoculum, normal-temperature flora and low-temperature flora on the straws respectively. The results are shown in FIGS. 5 and 6.
As can be seen from FIG. 5, the activities of the highest cellulase in 14 days of the microbial inoculum, the normal-temperature flora and the low-temperature flora are respectively 62.15U/mL, 56.31U/mL and 3.97U/mL; the 1 st, 3 rd, 5 th, 8 th and 14 th microbial inoculum has the same tendency of enzyme activity change of the endonuclease of the normal-temperature flora, but the microbial inoculum has higher enzyme activity than the endonuclease of the normal-temperature flora all the time; within 14 days, the cellulose endoenzyme activity of the cryophyte is always low, and the combination of the cryophyte and the normal temperature bacteria can promote or improve the cellulose enzyme activity of the normal temperature bacteria.
As can be seen from fig. 6, comparing 14 days, the residual amount of the straw converted by the original microbial inoculum, the normal temperature flora and the low temperature flora is almost unchanged, and the straw processed by the low temperature flora has better straw decomposition capability, although the original microbial inoculum is a combination of the normal temperature flora and the low temperature flora, the straw decomposition capability of the combined original microbial inoculum is not simply superposed by the decomposition capability of 2 floras, and although the low temperature flora is poor in the capability of decomposing macromolecular organic substances of the straw, the intermediate products of straw metabolism, such as disaccharides, acids, alcohols, lipids and the like, can be converted, so that the repression or inhibition of the metabolic products caused by the straw decomposition products is reduced or eliminated, and the straw decomposition is promoted.
Example 8
Effect of 4 different treatment modalities on product Activity
And (2) centrifuging the low-temperature flora prepared in the example 7 at 4000rpm for 5min to prepare a bacterial suspension with the bacterial content of 50%, calculating the number of bacterial colonies of the bacterial suspension by using a plate coating method, and adding the components in a mass ratio of 7: 1 (defatted powder: sodium ascorbate: 5: 1), and preparing the low-temperature bacteria containing the protective material.
Treatment 1: adding a carrier (5: 1 of corn flour and rice bran) into the low-temperature bacteria containing the protective material, wherein the mass ratio of the low-temperature bacteria to the carrier is 2: 1, serving as a common protective treatment microbial inoculum;
and (3) treatment 2: extruding the low-temperature bacteria containing the protective material and sodium alginate with the volume ratio of 2% drop by drop into a 2.5% calcium chloride solution by using a sterile injector to prepare sodium alginate microcapsules;
and (3) treatment: mixing low-temperature bacteria containing a protective material with 2% of sodium alginate and 2% of chitosan 1: 2, mixing, and extruding drop by drop into a 2.5 percent calcium chloride solution by using a sterile injector to prepare sodium alginate + chitosan microcapsules;
and (4) treatment: the bacterial suspension is prepared into freeze-dried powder by using a freeze-drying technology.
Meanwhile, the finished products treated in the above 4 ways are placed under the same conditions of room temperature (20-25 ℃) for 6 months, and samples are taken at 1, 2, 4, 5 and 6 months respectively to determine the survival rate of the cryophyte, and the results are shown in fig. 7: the 4 treatments (common protection, sodium alginate + chitosan and freeze-dried powder) have great influence on the survival rate of the low-temperature flora, and the survival rates of the microbial agents are sequentially from low to high by comparing the curves corresponding to the 4 treatments: the survival rates of common protection sodium alginate and chitosan freeze-dried powder are respectively 27.6%, 59.8%, 67.8% and 85.1% after 6 months of storage, and the results show that the freeze-dried powder prepared by freeze drying is more favorable for storage and use of low-temperature flora at room temperature, and more microorganisms survive and are favorable for maintaining the optimal activity of strains.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A cold region straw decomposition microbial inoculum is characterized by comprising a low-temperature flora and a normal-temperature flora; the low-temperature flora comprises lactobacillus plantarum with the cell number content of 20-28%, lactococcus lactis subspecies lactis with the cell number content of 17-25%, lactococcus 14-22%, arthrobacter globiformis with the cell number content of 11-22% and rhodococcus 9-18%; the normal-temperature flora is lignocellulose decomposition flora.
2. The cold region straw decomposition microbial inoculum according to claim 1, wherein the low temperature flora is prepared into freeze-dried powder or is embedded in a microcapsule carrier.
3. The cold-region straw decomposing microbial inoculum according to claim 2, wherein the freeze-dried powder or the microcapsule carrier comprises a cryo-protectant, a cryo-antioxidant; the low-temperature bacterium protective agent comprises one or more of skim milk powder, alpha-glucopyranose, NaCl, glycerol, lactose, polyethylene glycol and trehalose; the low-temperature bacterium antioxidant comprises one or more of vitamin E, tocopherol, cysteine hydrochloride, sodium ascorbate, tert-butyl hydroquinone, ascorbyl palmitate, isoascorbic acid and sodium erythorbate; the mass ratio of the low-temperature bacterium protective agent to the low-temperature bacterium antioxidant is 3-20: 1; the mass of the low-temperature flora thallus is 6-8 times of the sum of the mass of the low-temperature flora protective agent and the mass of the low-temperature flora antioxidant.
4. The cold-region straw decomposing microbial inoculum according to claim 3, wherein the microcapsule carrier further comprises a base material and a coagulant; the matrix material comprises sodium alginate and/or chitosan; the coagulant comprises CaCl2。
5. The cold-region straw decomposing microbial inoculum according to claim 1, wherein the normal-temperature flora comprises 20-27% of aspergillus oryzae, 17-24% of penicillium oxalicum, 14-21% of trichoderma viride, 3-9% of penicillium decumbens, 11-18% of bacillus subtilis, 8-15% of clostridium cellulolyticum, 5-12% of bacillus cereus, 3-5% of saccharomyces cerevisiae and 3-5% of pichia guilliermondii.
6. The cold-region straw decomposing microbial inoculum according to claim 5, wherein the normal-temperature flora is mixed with a normal-temperature bacteria protective agent and a normal-temperature bacteria antioxidant to prepare normal-temperature flora bacterial powder; the normal temperature bacteria protective agent comprises one or more of glycerol, polyethylene glycol, peptone, tween, trehalose, anhydrous sodium acetate, dipotassium hydrogen phosphate, magnesium sulfate and manganese sulfate; the room temperature bacteria antioxidant comprises one or more of shiitake mushroom scrap, white mushroom scrap, Chelidonium majus scrap, waste tea powder, Salvia miltiorrhiza powder, licorice scrap, clove scrap, monkshood powder, gallic acid powder and superoxide dismutase SOD; the mass ratio of the room-temperature bacterium protective agent to the room-temperature bacterium antioxidant is 1 (1-10); the mass of the normal-temperature flora thallus is 6-7 times of the sum of the mass of the normal-temperature flora protective agent and the mass of the normal-temperature flora antioxidant.
7. The cold-region straw decomposing microbial inoculum according to claim 1, wherein the cell number content ratio of the low-temperature flora to the normal-temperature flora is 1 (3-6).
8. The method for preparing the cold-region straw decomposing microbial inoculum according to any one of claims 1 to 7, which is characterized by comprising the step of flora culture, wherein the flora culture comprises low-temperature flora culture and normal-temperature flora culture; the culture medium for low-temperature flora culture comprises 5-25 g/L of carbon source and K2HPO4 0.8～1.2g/L，KH2PO40.8-1.2 g/L, 2-10 g/L peptone, 2-10 g/L beef extract, 1-5 g/L, MgSO yeast powder4·7H20.08-0.12 g/L of O, 1.8-2.2 g/L of sodium acetate and 1.8-2.2 g/L, MnSO of diammonium hydrogen citrate4·H20.04-0.06 g/L of O and 800.8-1.2 mL/L of Tween; the carbon source comprises glucose and/or sucrose; the culture medium for culturing the normal-temperature flora comprises (NH)4)2SO4 1.2～1.6g/L，MgSO4·7H2O 0.2～0.4g/L，KH2PO4 1.8～2.2g/L，CaCO31.8-2.2 g/L, peptone 2.2-2.8 g/L, FeSO4·7H2O 4～6mg/L，MnSO4 1.2～2mg/L，ZnCl2 1.4～2mg/L，CoCl21.4-2 mg/L and 6-10 g/L of straw.
9. The application of the cold-region straw decomposition microbial inoculum according to any one of claims 1 to 7 and the cold-region straw decomposition microbial inoculum prepared by the preparation method according to claim 8 in straw decomposition.
10. The application according to claim 9, wherein the application comprises: the cold-region straw decomposition microbial inoculum is applied to straw in-situ returning, and the application amount of the cold-region straw decomposition microbial inoculum is 0.5-1.2 kg/mu.
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Effective date of registration: 20231213 Address after: Room 617, Science and Technology Innovation Center Building, No. 335 Jiansanjiang Yingbin Road, Fujin City, Jiamusi City, Heilongjiang Province, 154000 Patentee after: Heilongjiang Zhongke Futian Straw Returning Technology Co.,Ltd. Address before: 150040 No. 26 Hexing Road, Xiangfang District, Heilongjiang, Harbin Patentee before: NORTHEAST FORESTRY University |