CN113684146B - Ultrahigh-temperature decomposed microbial agent and preparation method and application thereof - Google Patents
Ultrahigh-temperature decomposed microbial agent and preparation method and application thereof Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Abstract
The invention belongs to the field of microorganisms, and particularly relates to an ultrahigh temperature decomposed microbial agent, and a preparation method and application thereof. The specific technical scheme is as follows: the bacillus urealyticus is preserved in China general microbiological culture Collection center (China Committee) for culture Collection of microorganisms at a preservation address of 2021, 3 and 29 days: the preservation number of the Beijing city Chaoyang area North Chen Xili No. 1 and 3 is: CGMCC No.22086. And based on the bacillus urealyticus, a decomposing inoculant is prepared, and further comprises geobacillus, an accelerator and ferment. According to the invention, two new high-temperature resistant microorganisms are screened out, and the aerobic fermentation temperature can be increased to more than 70 ℃ by adding the extreme thermophilic microorganisms into the compost raw material without external heating, so that the fermentation production period is effectively shortened, a new way is provided for efficiently treating the organic solid waste, and the treatment technical level of harmless treatment, reduction and recycling of the organic solid waste is improved.
Description
Technical Field
The invention belongs to the field of microorganisms, and particularly relates to an ultrahigh temperature decomposed microbial agent, and a preparation method and application thereof.
Background
The problem of organic solid waste treatment is an important content of ecological environment protection and treatment. Organic solid waste includes crop waste, livestock manure, household garbage, sludge, and the like; the livestock and poultry raising waste produced by China every year is about 40 hundred million tons, and the comprehensive utilization rate is less than 60%; annual yield of straw is approximately 9 hundred million tons, and the unutilized part is up to 2 hundred million tons; annual urban sludge exceeds 7 millions of tons, and a large amount of sludge is discharged without proper treatment, so that secondary pollution of underground water, surface water, soil, air and the like is caused. The organic solid wastes are large in quantity and wide in range, are randomly placed in a disordered way and are randomly burnt, so that the ecological environment is seriously damaged.
High-temperature aerobic composting is one of the important ways to treat organic solid wastes. However, the traditional high-temperature composting technology has the defects of low fermentation temperature, long period, incomplete harmless treatment, serious odor pollution and the like, and severely limits the comprehensive popularization and application of the technology. Patent CN 109679857A discloses a compost decomposing inoculant, which is prepared by uniformly mixing the inoculant with auxiliary materials, inoculating the mixture into a compost material, and fermenting at 55-65 ℃. The fermentation temperature in the process is low, harmful bacteria which are not killed possibly remain, and external temperature control is needed, so that the operation is troublesome. Patent CN 102660479B discloses a composting fermentation composite microbial agent, a solid state fermentation production method and application thereof, the fermentation completion time is 25-30 days, the decomposition efficiency is low, the preparation process of the microbial agent is complex, and the operation is not easy.
Therefore, a microbial inoculum for ultra-high temperature decomposition is needed to improve the treatment efficiency and effect of organic solid wastes.
Drawings
FIG. 1 is a colony morphology diagram of the ureido-degrading bacillus provided by the invention;
FIG. 2 is a microscopic view of the Bacillus urealyticum provided by the invention;
FIG. 3 is a morphology of geobacillus colonies provided by the present invention;
fig. 4 is a microscopic view of geobacillus provided by the invention.
Disclosure of Invention
The invention aims to provide an ultrahigh temperature decomposed microbial agent, and a preparation method and application thereof.
In order to achieve the aim of the invention, the invention adopts the following technical scheme: bacillus urealyticus (Ureibacillus sp.) was stored in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) at a storage address of 2021, 3 months: the preservation number of the Beijing city Chaoyang area North Chen Xili No. 1 and 3 is: CGMCC No.22086.
Correspondingly, a strain of bacillus urealyticus (Ureibacillus sp.) has a 16S rDNA sequence shown in SEQ ID NO 1.
Correspondingly, the application of the bacillus urealyticum in composting is provided.
Correspondingly, the composting microbial inoculum comprising the bacillus urealyticus.
Preferably, the microbial inoculum further comprises Geobacillus sp.
Preferably, the geobacillus is stored in China general microbiological culture Collection center (CGMCC) at the 3 rd month 29 of 2021, and the storage address is: the preservation number of the Beijing city Chaoyang area North Chen Xili No. 1 and 3 is: CGMCC No.22088, or the 16S rDNA sequence of the geobacillus is shown as SEQ ID NO 2.
Preferably, in the microbial inoculum, the viable bacteria ratio of the ureido bacillus to the geobacillus is 2:1-1:1.
Preferably, the ratio of the viable bacteria is 1:1.
Preferably, the microbial inoculum further comprises ferment.
Preferably, when the microbial inoculum is a solid microbial inoculum, the microbial inoculum further comprises an accelerator.
The invention has the following beneficial effects: the invention aims to develop the ultra-high temperature aerobic fermentation composite microbial inoculum through research and improve the composting efficiency and composting quality of organic solid wastes. By adding extreme thermophilic microorganisms into the compost raw materials, the aerobic fermentation temperature can be increased to be more than 70 ℃ under the condition of no external heating, so that the fermentation production period is effectively shortened, a new way is provided for efficiently treating the organic solid waste, and the treatment technical level of harmless treatment, reduction and recycling of the organic solid waste is improved.
Detailed Description
The invention provides an ultra-high temperature decomposing inoculant for composting, which comprises bacillus urealyticus (hereinafter referred to as X501) and Geobacillus sp (hereinafter referred to as X701).
Preferably, the microbial inoculum further comprises ferment.
The preferable scheme is as follows: in the microbial inoculum, the viable bacteria ratio of the bacillus urealyticus to the geobacillus is 2:1-1:1; more preferably, the viable bacteria ratio is 1:1. In the microbial inoculum, the effective viable count of the bacillus urealyticus and the geobacillus denitrificans is more than or equal to 6 multiplied by 10 9 Each/g.
The preferable scheme is as follows: the ferment accounts for 10-15% of the volume of the microbial inoculum.
The preferable scheme is as follows: the ferment is fruit ferment and is obtained by mixing and fermenting fruits or pericarps which are not easy to spoil and have fruit fragrance, such as apples, pears, orange peel, shaddock peel and the like, with one or more probiotics, such as lactobacillus plantarum, aroma-producing yeast and the like.
The more preferable scheme is as follows: the preparation method of the ferment comprises the following steps: according to the mass ratio, water and fruit are mixed uniformly, and the mixture is filled into a fermentation barrel, wherein the sample filling amount is 80-85% of the volume of the fermentation barrel, and meanwhile, the brown sugar is added after being melted. Inoculating lactobacillus plantarum, aroma-producing yeast and other probiotics bacterial liquid, preferably adding lactobacillus plantarum and aroma-producing yeast simultaneously, and adding lactobacillus plantarum bacterial liquid and aroma-producing yeast bacterial liquidThe amount of the active bacteria is 0.5 to 1 percent of the volume of the fermentation substrate, and the concentration of the active bacteria in the bacterial liquid is more than or equal to 10 8 And each mL. Stirring, spreading plastic film, and sealing. And (3) extracting ferment water from the fermented mixed bacterial liquid after 3 months, and filtering to obtain the required ferment.
The invention also provides a preparation process of the ultra-high temperature decomposed microbial inoculum. The microbial inoculum can be prepared into a liquid microbial inoculum or a solid microbial inoculum according to actual needs.
The preparation method of the liquid microbial inoculum specifically comprises the following steps:
1. x501 and X701 are respectively connected into corresponding solid culture mediums (beef extract peptone culture medium or LB culture medium, and beef extract peptone culture medium is more preferable) and activated for standby. The culture temperature of the ureidobacillus urealyticus X501 is 35-60 ℃, and the culture temperature of the geobacillus X701 is 40-75 ℃.
2. And (3) inoculating the activated strains into sterilized liquid culture media (beef extract peptone culture media or LB culture media, and beef extract peptone culture media are more preferable) for culture, wherein the dissolved oxygen of the fermentation liquid is 0.1-0.5 mg/L, and the fermentation time is 24-48 h.
3. The count of the two cultured bacterial liquids is regulated to be 10 for the effective viable bacteria 9 CFU/mL, and then uniformly mixing the two bacterial solutions according to a proportion to obtain the liquid composite microbial agent.
4. And adding the ferment filtrate, and uniformly mixing to obtain the liquid ultrahigh-temperature decomposed microbial inoculum.
The preparation method of the solid microbial inoculum specifically comprises the following steps:
1. sterilizing bran to obtain adsorbent. And adding 1Kg of adsorbent into each liter of the liquid ultra-high temperature decomposed microbial inoculum, and stirring and uniformly mixing.
2. Adding the promoter according to the proportion of 5g of promoter in each kilogram of microbial inoculum, and uniformly stirring to promote the activation of microorganisms. The promoter is preferably yeast extract, and commercially available yeast powder can be used directly. When solid microbial agents are used, the strain needs to be activated. The promoter may provide a promoting effect upon activation of the strain. The activity of the strain in the liquid microbial inoculum is relatively high, and the activation can be assisted without adding an accelerator alone.
3. Air-drying at 30-40 deg.C for 2-3 d until the water content is 15-20%. And then crushing the mixture to the required mesh number by a crusher, and quantitatively packaging and preserving the mixture at a low temperature (4-20 ℃) to obtain the solid composite ultrahigh-temperature decomposed microbial inoculum.
In the following, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments. The technical means used in the examples are conventional means well known to those skilled in the art unless otherwise indicated.
Embodiment one: screening, identification and preservation of microorganisms
1. Screening: 10g of a fresh sludge compost sample obtained after high-temperature fermentation is weighed, placed in a triangular flask filled with glass beads and 90mL of sterile water, placed in a constant-temperature shaking table at 30 ℃ for 30min, and then the suspension (10 is sucked up -1 ) 10mL of the mixture was placed in a triangular flask containing 90mL of sterile water according to a ratio of 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 、10 -8 、10 -9 And (3) carrying out gradient dilution on the compost sample suspension. Samples with different dilutions are respectively coated on a plate of a separation solid culture medium, and are inversely cultured in an incubator at 50 ℃ and 70 ℃ for 1-2 days. All single colonies with different characteristics are picked, streaked and separated on a solid flat plate culture medium, and the culture is repeated for 3 times, so that the high-temperature resistant bacteria pure strain is obtained and numbered. The culture medium used for separation and purification is beef extract peptone agar culture medium, wherein beef extract is 3.0g/L, peptone is 10.0g/L, sodium chloride is 5.0g/L, agar powder is 20g/L, pH is adjusted to 7.2-7.4, and the beef extract peptone agar culture medium is sterilized for 30min at 121 ℃ in an autoclave.
Selecting the strain with the best performance in the microorganism obtained by separation and being most suitable for high-temperature composting, and carrying out 16S rDNA sequencing comparison and identification. The identification strain X501 is bacillus urealyticus, the 16S rDNA sequence is shown as SEQ ID NO 1, the colony morphology is shown as figure 1, the colony is smaller, opaque, yellowish, dry and flat and easy to pick, and the microscopic examination chart is shown as figure 2. The temperature range of the bacillus urealyticum is 30-55 ℃ and the optimal growth temperature is 50 ℃. The pH is suitably in the range of 5 to 9, and the most suitable pH is 7.0. The carbon source includes corn starch, potato starch, industrial glucose, etc., and the nitrogen source includes corn steep liquor, bean cake powder, yeast extract, etc.
The identification X701 is Geobacillus, the 16S rDNA sequence is shown as SEQ ID NO 2, the colony morphology is shown as figure 3, the colony size is different, the surface is white and rough and opaque, and the microscopic examination chart is shown as figure 4. The temperature range of the geobacillus is 40-75 ℃ and the optimal growth temperature is 60 ℃. The pH is suitably in the range of 5 to 9, and the most suitable pH is 7.0. The carbon source includes corn starch, potato starch, industrial glucose, etc., and the nitrogen source includes corn steep liquor, bean cake powder, yeast extract, etc.
Bacillus urealyticus (Ureibacillus sp.) X501 is stored in China general microbiological culture Collection center (CGMCC) at 2021, 3 and 29 days, and the storage address is: the preservation number of the Beijing city Chaoyang area North Chen Xili No. 1 and 3 is: CGMCC No.22086. Geobacillus (Geobacillus sp.) X701 is preserved in China general microbiological culture Collection center (CGMCC) at 2021, 3 and 29 days, and the preservation address is: the preservation number of the Beijing city Chaoyang area North Chen Xili No. 1 and 3 is: CGMCC No.22088. Note that, pull Ding Ming at the naming of the division in the preservation is simply referred to, and the other places are fully referred to herein.
Embodiment two: preparation and application of solid composite super-temperature decomposed microbial inoculum
1. Quantitatively using a beef extract peptone solid culture medium subjected to high-temperature sterilization treatment (121 ℃ for 20 minutes) for later use, inoculating a strain X501 into the beef extract peptone solid slant culture medium subjected to sterilization treatment, and placing the beef extract peptone solid slant culture medium in a 50 ℃ incubator for later use for culture and activation. Quantitatively using a beef extract peptone solid culture medium subjected to high-temperature sterilization treatment (121 ℃ for 20 minutes) for later use, inoculating a strain X701 into the beef extract peptone solid slant culture medium subjected to sterilization treatment, and placing the beef extract peptone solid slant culture medium in a 60 ℃ incubator for later use for culture and activation.
The beef extract peptone liquid culture medium subjected to high-temperature sterilization treatment (121 ℃ for 20 minutes) is quantitatively reserved, and activated strain X501 is inoculated into the beef extract peptone liquid culture medium and is cultured for 24 hours by shaking (50 ℃ for 160 revolutions per minute). The beef extract peptone liquid culture medium subjected to high-temperature sterilization treatment (121 ℃ for 20 minutes) is quantitatively reserved, and then activated strain X701 is inoculated into the beef extract peptone liquid culture medium and is cultured for 24 hours by shaking (60 ℃ for 160 revolutions per minute).
Counting the cultured bacterial liquid, and regulating the effective viable count of each strain to be 10 9 CFU/mL, 19 groups of mixed bacterial solutions were prepared in the manner of combination in Table 1. And adding ferment filtrate according to the mode of the table 1 to obtain the liquid ultra-high temperature decomposed microbial inoculum. The preparation method of the ferment filtrate comprises the following steps: mixing water and apple with brown sugar=10:3:1, loading into a fermentation barrel, cleaning and dicing, wherein the total loading amount of the water and the apple is 85% of the volume of a fermentation pit, simultaneously adding the melted brown sugar, and adding 1% (v/v) lactobacillus plantarum and 1% (v/v) aroma-producing yeast into the fermentation barrel, wherein the viable bacteria concentration of the lactobacillus plantarum and the aroma-producing yeast is 10 respectively 8 And each mL. Stirring uniformly, spreading a plastic film, covering, sealing, fermenting for 3 months, and extracting fermentation liquor after 3 months, wherein the filtered liquor is ferment filtrate.
According to the proportion shown in Table 1, the liquid ultra-high temperature decomposed microbial inoculum is added into the sterilized bran. Selecting an open and cool place, and air-drying the composite microbial inoculum for 3d at 35 ℃ until the water content is 15-20%. Adding promoter (commercially available yeast powder) according to the mode of Table 1, stirring uniformly, pulverizing by a pulverizer to obtain solid ultra-high temperature decomposed microbial inoculum, and packaging and storing in a cool and dry low-temperature environment at 4-20deg.C.
In Table 1, the mixed bacterial liquid is the fermentation liquid of the single microorganism in the group to which only one microorganism is added. The commercial ureido bacillus is purchased from the German collection of microorganisms and cell cultures, with the deposit number DSM17952; the commercial Geobacillus is purchased from Beijing North Innovative biological technology institute, model BNCC161884.
Table 1 shows the preparation parameters of each group of solid ultra-high temperature decomposed microbial inoculants
2. The prepared solid ultra-high temperature decomposed microbial inoculum of each group is used for garden garbage composting. And (3) sorting garden garbage (road fallen leaves, lawn trimmings, garden pruning and the like), removing non-biological material foreign matters such as plastics, stones and the like, and crushing the sorted garden garbage to the grain size of 2cm. The solid ultra-high temperature decomposed microbial inoculum of each group is inoculated into a garden garbage pile body according to the inoculation quantity (mass ratio) of 0.1%, and the length, width and height of the pile body are respectively 3m, 2m and 1m. C/N is adjusted to 20-30:1, preferably C/N and C/N of the embodiment is adjusted to 25:1 (specifically, when C/N is lower, crushed wheat straw or rice straw or corn straw is added, and when C/N is higher, urea is added), meanwhile, slaked lime is added, pH and water content are simultaneously adjusted, and the water content of garden garbage is controlled to be 61%. The method for adding the slaked lime comprises the following steps: if the water content of the garden garbage raw material is less than or equal to 55%, adding the slaked limestone ash water in the form of solution; if the water content of the garden garbage is more than 55%, adding slaked lime solid. And then standing for composting fermentation.
In the fermentation process, pile turning, aeration and water supplementing are carried out to control the condition factors in the fermentation process, namely temperature, ventilation and water content, and the method specifically comprises the following steps: the water content is controlled to be 55-65%, water is dripped by hand grasping the hand seam, and the specific measurement method is measured according to the method of poor drying quality. Aerating and supplying oxygen to the materials by a high-pressure fan, an air compressor or a stirring mode; the ventilation mode using the air compressor is as follows: aeration was performed intermittently at a ventilation rate of 1.8L for 3 hours per day at half an hour intervals. When the temperature is maintained at about 65 ℃ for 3-4 d, the pile is turned manually, naturally ventilated and supplied with oxygen, water is added, and the mixture is stirred uniformly. And (3) when the composting fermentation is completed and the composting temperature is reduced to below 40 ℃ and the moisture is reduced to below 30 percent, and when the (terminal point C/N)/(initial C/N) is smaller than 0.6, the composting fermentation is completed, and finally screening is carried out to obtain the finished organic fertilizer product.
Except for each treatment of the different solid ultra-high temperature decomposed microbial inoculum settingsA blank control group (CK) was also set at the same time: no microbial agent is added, equal amount of bran is inoculated for replacement, and the rest conditions are the same; positive control group: 0.1% of a commercial decomposition agent was added, the remaining conditions being the same. The commercial organic fertilizer fermentation inoculant contains bacillus subtilis 2 multiplied by 10 10 CFU/g, bacillus amyloliquefaciens 2×10 10 CFU/g. The change in stack temperature was measured daily for each group and samples were taken periodically (mixed using multipoint sampling). And determining the microbial mass, organic matters, total nitrogen, total potassium, total phosphorus and seed germination rate of the sample.
The temperature measuring method comprises the following steps: a temperature probe (the insertion depth is 40-50 cm) is inserted into the central line of the pile body, the temperature change in the pile body in the composting process is monitored, and the temperature at nine am points of each day is taken as a measured value.
The method for measuring the microorganism content (bacterial number) is a dilution plate counting method, and a beef extract peptone culture medium is adopted as a culture medium.
The organic matter content is determined with reference to appendix C in the national standard NY 525-2021. The total content of nitrogen, phosphorus and potassium is determined by adopting annex D in national standard NY 525-2021. The germination index of the seeds after the decomposition (after the composting) is measured by referring to annex F in the national standard NY 525-2021, and the germination index is measured after the composting.
The results of each group are shown in tables 2, 3 and 4.
TABLE 2 comparison of composting effect (temperature) of solid ultra-high temperature decomposing inoculants of each group
TABLE 3 comparison of composting effects (organic matter and Properties) of various groups of solid ultra-high temperature decomposing inoculants
Table 4 comparison of composting effect (seed germination rate) of each group of solid ultra-high temperature decomposed microbial agents
| Group of | Seed germination rate (GI) |
| Group 1 | 88.09％ |
| Group 2 | 81.38％ |
| Group 3 | 90.10％ |
| Group 4 | 85.88％ |
| Group 5 | 94.67％ |
| Group 6 | 94.38％ |
| Group 7 | 93.29％ |
| Group 8 | 92.15％ |
| Group 9 | 91.57％ |
| Group 10 | 88.69％ |
| Group 11 | 86.25％ |
| Group 12 | 87.38％ |
| Group 13 | 87.12％ |
| Group 14 | 89.52％ |
| Group 15 | 93.29％ |
| Group 16 | 85.13％ |
| Group 17 | 87.21％ |
| Group 18 | 89.33％ |
| Group 19 | 88.15％ |
| Blank control | 69.54％ |
| Positive control | 85.61％ |
3. The prepared solid ultra-high temperature decomposed microbial inoculum of group 5, blank control group and positive control group is used for livestock manure composting. In a chicken farm in the Yangtze river area of the city of Sichuan province, inoculating each group of solid ultra-high temperature decomposed microbial inoculum into chicken manure piles according to the inoculation amount of 0.1%, adding rice bran slag to adjust the C/N to 25:1, controlling the water content of the compost to 59.8%, fully and uniformly mixing, and piling into a strip pile body with the width of 1.2m and the length of 2 m. The compost is turned once every 3 days in the initial stage of the compost, and the high-temperature stage (from the first temperature rise of the compost body to 50 ℃ until the temperature of the compost body is reduced to 50 ℃ and the temperature is not raised any more) in the middle stage of the compost is turned once every 5 days. A blank control group and a positive control group are also arranged in a mode of garden garbage compost. The results of each group are shown in Table 5.
TABLE 5 comparison of composting effects (organic matter and Properties) of various groups of solid ultra-high temperature decomposing inoculants
4. The prepared group 5 solid ultra-high temperature decomposed microbial inoculum is used for sludge composting, and a blank control group is reserved. Straw is used as auxiliary material, the C/N is adjusted to be 26:1 by adding straw, the water content is adjusted to be 61%, and the mixture is uniformly mixed, and the stack is a strip stack body with the width of 1.2m and the length of 2 m. And (3) inoculating solid ultra-high temperature decomposed microbial inoculum of each group according to the inoculation amount of 1%, and setting a blank control group and a positive control group according to a garden garbage composting method. After fully and uniformly mixing each group of sludge and the microbial inoculum, turning the pile once every 4 d.
The organic matter degradation rate (mass fraction of organic matter measured on a dry basis, performed according to annex C of national standard NY 525-2021), total nitrogen, and humus content (sodium pyrophosphate leaching-potassium dichromate capacity method) of each group of stacks were measured.
After the fermentation of the pile is completed, crushing, sieving, detecting, weighing and packaging are carried out, so that each group of organic fertilizer is prepared. And (3) carrying out field comparison experiments on a farmland experiment base in double-flow areas of the Chengdu city, wherein the tested crops are commercially available cabbages. According to the habit of applying local fertilizer, the fertilizer and fine soil are uniformly mixed and then placed into each hole by a hole application method, and then the seedlings of the Chinese cabbage are transplanted. Matching withCommercial fertilizer (50 Kg/mu, potassium sulfate type "vegetable, fruit and vegetable compound fertilizer" purchased from Taobao Tai mountain ecological agriculture store) was applied as a blank group 2, and then an equal amount of each group of organic fertilizer was applied, and three replicates were set for each group (i.e., 3 patch areas, each patch area planted with 50 plantlets). During the test, the treatments are consistent with other field management measures except for different fertilization. After one month, 40 cabbages are randomly selected from each area to measure the fresh weight of each plant, and the plant diseases and insect pests are observed. And after the measurement is finished, harvesting in a partition, and counting the actual product. The results of each group are shown in Table 6. Wherein, the average incidence (%) of chinese cabbage=the number of plants developed in all blocks/the total number of plants in all blocks×100%. Relative control (%) = (control disease index-treatment disease index)/control disease index x 100%. The disease index = Σ (number of disease stages at each stage x disease stage)/(total number of investigation x highest representative value) ×100% was recorded while investigating the disease rate of the plants by referring to the investigation grading standard of the disease index of tomato late blight (DB 51T 1652-2013 technical specification for tomato late blight). The grading method comprises the following steps: grade 0, no lesions; 1 grade, the area of the disease spots accounts for less than 25% of the whole leaf area; the area of the lesion accounts for 26% -50% of the whole leaf area; stage 3, the area of the lesion accounts for 51% -75% of the whole leaf area; grade 4, the area of the disease spots accounts for more than 76% of the whole leaf area or the plants die. Per mu yield fresh weight = block yield x 666/block area; the term 666 is 1 mu= 666.7m 2 Take value 666 for ease of calculation. Yield increase= (total treatment group yield-total blank group yield)/total blank group yield x 100%.
Table 6 comparison of composting effects of groups of solid ultra-high temperature decomposing inoculants
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications, variations, alterations, substitutions made by those skilled in the art to the technical solution of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the spirit of the design of the present invention.
Sequence listing
<110> institute of biological research for adult and capital of academy of sciences in China
Sichuan green microorganism technology Co.Ltd
<120> an ultra-high temperature decomposed microbial inoculum, and preparation method and application thereof
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cttcggtgtg ttatccggta ttagcacgcg tttccgcgag ttatcccggt ctatagggca 1320
ggttacccac gtgttactca cccgtccgcc gctaaccaat taaaaagcaa gcttctaatt 1380
gtccgctcga ctg 1393
<210> 2
<211> 1456
<212> DNA
<213> Geobacillus thermodenitrificans
<400> 2
gctggcggtg ctataatgca gtcgagcgga ccgaacgaga gcttgctctt gtttggtcag 60
cggcggacgg gtgagtaaca cgtgggcaac ctgcccgcaa gaccgggata actccgggaa 120
accggagcta ataccggata acaccaaaga ccgcatggtc tttggttgaa aggcggcttc 180
ggctgccact tgcggatggg cccgcggcgc attagctagt tggtgaggta acggctcacc 240
aaggcgacga tgcgtagccg gcctgagagg gtgaccggcc acactgggac tgagacacgg 300
cccagactcc tacgggaggc agcagtaggg aatcttccgc aatggacgaa agtctgacgg 360
agcgacgccg cgtgagcgaa gaaggccttc gggtcgtaaa gctctgttgt gagggacgaa 420
ggagcgccgt ttgaataagg cggcgcggtg acggtacctc acgagaaagc cccggctaac 480
tacgtgccag cagccgcggt aatacgtagg gggcgagcgt tgtccggaat tattgggcgt 540
aaagcgcgcg caggcggtcc tttaagtctg atgtgaaagc ccacggctca accgtggagg 600
gtcattggaa actgggggac ttgagtgcag gagaggagag cggaattcca cgtgtagcgg 660
tgaaatgcgt agagatgtgg aggaacacca gtggcgaagg cggctctctg gcctgtaact 720
gacgctgagg cgcgaaagcg tggggagcaa acaggattag ataccctggt agtccacgcc 780
gtaaacgatg agtgctaagt gttagagggg tcacaccctt tagtgctgta gctaacgcga 840
taagcactcc gcctggggag tacggccgca aggctgaaac tcaaaggaat tgacgggggc 900
ccgcacaagc ggtggagcat gtggtttaat tcgaagcaac gcgaagaacc ttaccaggtc 960
ttgacatccc ctgacaaccc aagagattgg gcgttccccc ttcgggggga cagggtgaca 1020
ggtggtgcat ggttgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag 1080
cgcaaccctt gcctctagtt gccagcattc agttgggcac tctagaggga ctgccggcta 1140
aaagtcggag gaaggtgggg atgacgtcaa atcatcatgc cccttatgac ctgggctaca 1200
cacgtgctac aatgggcggt acaaagggct gcgaacccgc gagggggagc gaatcccaaa 1260
aagccgctct cagttcggat tgcaggctgc aactcgcctg catgaagccg gaatcgctag 1320
taatcgcgga tcagcatgcc gcggtgaata cgttcccggg ccttgtacac accgcccgtc 1380
acaccacgag agcttgcaac acccgaagtc ggtgaggtaa cccttacggg agccagccgc 1440
cgaaggtgat caagtg 1456
Claims (4)
1. Bacillus urealyticus strainUreibacillussp.), characterized in that: the strain is preserved in China general microbiological culture Collection center (China Committee) for culture Collection of microorganisms (China) for type culture Collection of microorganisms (2021, 3, 29)The preservation address is: the preservation number of the Beijing city Chaoyang area North Chen Xili No. 1 and 3 is: cgmccno.22086.
2. A composting microbial inoculant comprising the bacillus urealyticus of claim 1, wherein: the microbial inoculum also comprises geobacillusGeobacillussp.) the Geobacillus was stored in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) at a storage address of 2021, 3 months and 29 days: the preservation number of the Beijing city Chaoyang area North Chen Xili No. 1 and 3 is: cgmccno.22088.
3. The composting microbial inoculum according to claim 2, characterized in that: in the microbial inoculum, the viable bacteria ratio of the bacillus urealyticus to the geobacillus is 2:1-1:1.
4. A composting microbial agent as claimed in claim 3 characterised in that: the ratio of the viable bacteria is 1:1.
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