CN106701621B - Preparation method of straw decomposition agent for decomposing at low temperature - Google Patents
Preparation method of straw decomposition agent for decomposing at low temperature Download PDFInfo
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Abstract
The invention relates to a compound microbial agent and a preparation method thereof, and solves the problems of slow decomposition starting and incomplete decomposition of a decomposition agent in the prior art. The invention has the beneficial effects that: 1. the straw decomposing inoculant disclosed by the invention uses the strain YO-H, so that the straw decomposing inoculant can be quickly decomposed under the condition of relatively low environmental temperature; 2. the strain YO-H is matched with other rotten bacteria, so that the rotten effect is better; 3. the additional enzyme preparation is added, so that the cellulase activity of the decomposing agent is improved, the additional enzyme preparation is utilized to degrade lignin and cellulose within 3-4 days before the decomposing bacteria are not propagated in large quantities to generate new enzymes, and the decomposing time is advanced by 3-4 days.
Description
The application is divided into separate applications by taking an invention patent with the application date of 2013-12-03 and the application number of 201310641595.4 and named as a straw decomposing agent capable of quickly starting to decompose at low temperature and a preparation method thereof as a parent.
Technical Field
The invention relates to a straw decomposing agent and a preparation method thereof, in particular to a straw decomposing agent for decomposing at a low temperature and a preparation method thereof.
Background
The organic materials such as the straws and the like contain a large amount of lignin, the lignin surrounds the hemicellulose and is filled between the microfibrils, the hemicellulose and the microfibril are tightly combined with cell wall components, and the lignin, the cellulose and the hemicellulose form a whole, so that the decomposition difficulty of the straws by microorganisms is increased. The straws are in the growth and propagation period of microorganisms in the decomposing agent in a few days before the straws are decomposed, the contribution to the decomposition is small, and only when the microorganisms are propagated and secrete various lignin and hemicellulose degrading enzymes in a large quantity, the tight combination between the lignin and the hemicellulose is damaged, the internal cellulose structure is exposed, and the straws enter the rapid decomposition stage. Often, the quantity of the 'three-element' enzyme secreted by the microorganisms is insufficient, and the activity is insufficient, so that the decomposition speed and degree of the straw are influenced.
The organic material decomposing inoculant breeds a large amount of decomposing bacteria at a proper environmental temperature, decomposes decomposed organic materials by a multienzyme system generated by metabolism activity of the organic material decomposing inoculant, and the conventional decomposing inoculant generally requires the environmental temperature for starting decomposition to be over 15 ℃, and is slow in starting decomposition, long in time and incomplete in decomposition at a lower temperature. The crop straws are generally idle in the ground after the harvest in 11 months in China is finished, and the composting and the maturity are generally carried out in the season. The temperature is lower in most areas in China in the season, and proper decomposition starting environment temperature is difficult to achieve.
Disclosure of Invention
In order to overcome the problems of slow decomposition starting and incomplete decomposition of the decomposing agent in the prior art, the invention provides the decomposing agent capable of quickly starting decomposition at a low temperature (lower than 10 ℃).
The technical scheme of the invention is to provide a straw decomposing agent for quickly starting decomposition at low temperature, the straw decomposing agent comprises arthrobacter strain YO-H, the strain YO-H is preserved in China general microbiological culture Collection center, and the preservation unit addresses are as follows: beijing, Chaoyang district Beichen Lu No. 1, institute of microbiology, academy of sciences of China, with a preservation date of 2010, 8.23.23.8.Latin, named Arthrobacter ariliati, and a strain name of "YO-H", with a preservation number of "CGMCC NO. 4119".
Preferably, the straw decomposition agent for rapidly starting decomposition at low temperature further comprises bacillus subtilis, saccharomyces cerevisiae, aspergillus niger and trichoderma harzianum.
Preferably, in the straw decomposition agent for rapidly starting decomposition at low temperature, the strain YO-H, the bacillus subtilis, the saccharomyces cerevisiae, the aspergillus niger and the trichoderma harzianum have a strain number ratio of 1: 1: 1: 2: 2.
the invention also provides a preparation method of the straw decomposing agent for quickly starting decomposition at low temperature, which comprises the following steps:
respectively culturing strain YO-H, Bacillus subtilis, Saccharomyces cerevisiae, Aspergillus niger and Trichoderma harzianum until the bacterial concentration is 108Per mL, in a volume ratio of 1: 1: 1: 2: 2, uniformly mixing to obtain a mixed bacterial liquid, inoculating the mixed bacterial liquid into wheat bran according to the proportion of 1:10, culturing for 10 days, wherein the mass fraction of the water in the wheat bran is 50%, air-drying the mixture in a shade until the mass fraction of the water in the wheat bran is less than 25%, adding cellulase with the mass fraction of 1% -1.5% of the wheat bran, and uniformly mixing to obtain the straw decomposition agent.
Preferably, the preparation method of the straw decomposition agent capable of quickly starting decomposition at low temperature comprises the following steps:
(1) carrying out enrichment culture on decaying corn straws at 5 ℃, coating a double-layer flat plate which takes cellulose as a unique carbon source after diluting an immersion liquid at 5 ℃ for culture until a single colony is formed, and puncturing and picking the colony to obtain a strain YO-H;
(2) respectively culturing strain YO-H, Bacillus subtilis, Saccharomyces cerevisiae, Aspergillus niger and Trichoderma harzianum until the bacterial concentration is 108Per mL, in a volume ratio of 1: 1: 1: 2: 2, uniformly mixing, inoculating the mixture into wheat bran according to the proportion of 1:10, culturing for 10 days, wherein the mass fraction of the water in the wheat bran is 50%, air-drying the mixture in a shade place until the mass fraction of the water in the wheat bran is less than or equal to 25%, adding cellulase with the mass fraction of 1% -1.5% of the wheat bran, and uniformly mixing to obtain the straw decomposition agent.
Preferably, in the above method for preparing a straw-decomposing inoculant capable of rapidly starting decomposition at a low temperature, the culture method of the strain YO-H is as follows:
1) activating strains: taking 1-ring strain YO-H lawn from a bevel storage tube into 30mL of liquid culture medium, and culturing at 37 ℃ and 200-250 rpm for 20-24 hours;
2) and (3) shake flask culture: drawing the bacterial liquid obtained in the step 1) into a shake flask by using a sterile pipettor, wherein the inoculation amount is as follows: 1 percent; temperature: 37 ℃; rotating speed: 200-250 rpm; culturing time: 20-24 h;
3) culturing in a fermentation tank: inoculating the shake flask cultured in the step 2) into a fermentation tank for amplification culture in an inoculation mode: flame inoculation; inoculation amount: 1 percent; temperature: 37 ℃; rotating speed: 180-200 rpm; the ventilation volume is 1: 0.8; and (3) tank pressure: 0.02 MPa; time: 24-36 h; obtaining strain YO-H bacterial liquid;
the method for activating the strain and preparing the culture medium for shaking the flask comprises the following steps: adding deionized water into bean sprouts, boiling for 20 minutes until the mass fraction of the bean sprouts is 25%, filtering to remove residues, supplementing the volume, adding 5% of glucose in mass fraction, dissolving and uniformly mixing, adjusting the pH value to 7.0, and sterilizing for 30 minutes at 121 ℃;
the preparation method of the culture medium for counting the strain inclined plane and the plate comprises the following steps: adding deionized water into bean sprouts, boiling for 20 minutes until the mass fraction of the bean sprouts is 25%, filtering and deslagging, complementing the volume, adding 5% of glucose and 2% of agar powder by mass fraction, dissolving and uniformly mixing, adjusting the pH value to 7.0, and sterilizing for 30 minutes at 121 ℃.
The invention has the beneficial effects that: 1. the straw decomposing inoculant disclosed by the invention uses the strain YO-H, so that the straw decomposing inoculant can be quickly decomposed under the condition of relatively low environmental temperature; 2. the strain YO-H is matched with other rotten bacteria, so that the rotten effect is better; 3. the additional enzyme preparation is added, so that the cellulase activity of the decomposing agent is improved, the additional enzyme preparation is utilized to degrade lignin and cellulose within 3-4 days before the decomposing bacteria are not propagated in large quantities to generate new enzymes, and the decomposing time is advanced by 3-4 days.
Detailed Description
In order to explain technical contents, structural features, and objects and effects of the present invention in detail, the following description is given in detail with reference to the embodiments.
The embodiment of the invention is as follows:
example 1
Performing enrichment culture on decaying corn straws at a low temperature (5 ℃), diluting the infusion, selecting a proper dilution to coat a double-layer flat plate with cellulose as a unique carbon source at the low temperature (5 ℃) for culture until a single bacterial colony is formed, and puncturing and picking the bacterial colony to obtain the low-temperature rotten bacteria. Identified as arthrobacterium YO-H with the preservation number of CGMCC NO. 4119.
Respectively culturing strain YO-H, Bacillus subtilis, Saccharomyces cerevisiae, Aspergillus niger and Trichoderma harzianum until the bacterial concentration is 108Per mL, in a volume ratio of 1: 1: 1: 2: 2, uniformly mixing, inoculating the mixture into wheat bran with 50% of water content according to the proportion of 1:10, culturing for 10 days, airing the mixture in a shade until the mass fraction of the water content in the wheat bran is less than or equal to 25%, adding 1% -1.5% of cellulase, and uniformly mixing to obtain the straw decomposition agent.
The culture method of the arthrobacter strain YO-H comprises the following steps:
shake flask and fermentation medium for strain: liquid medium. Adding 75 parts of deionized water into 25 parts of bean sprouts by weight, boiling for 20 minutes, filtering to remove residues, supplementing the volume, adding 5 parts of glucose by weight, dissolving and uniformly mixing, adjusting the pH value to 7.0, and sterilizing for 30 minutes at 121 ℃.
Culture medium for slant and plate counting of strains: solid medium. Adding 75 parts by weight of deionized water into 25 parts by weight of bean sprouts, boiling for 20 minutes, filtering to remove residues, supplementing the volume, adding 5 parts by weight of glucose and 2 parts by weight of agar powder, dissolving and uniformly mixing, adjusting the pH value to 7.0, and sterilizing for 30 minutes at 121 ℃.
Activating strains: taking 1-ring lawn from the slant preservation tube to 30mL liquid culture medium, culturing at 37 deg.C and 200-250 rpm for 20-24 hours.
And (3) shake flask culture: the activated bacteria liquid is drawn into a shake flask by a sterile pipette. Inoculation amount: 1 percent; temperature: 37 ℃; rotating speed: 200-250 rpm; culturing time: 20-24 h.
Culturing in a fermentation tank: inoculating the cultured shake flask into a fermentation tank for enlarged culture. An inoculation mode comprises the following steps: flame inoculation; inoculation amount: 1 percent; temperature: 37 ℃; rotating speed: 180-200 rpm; the ventilation volume is 1: 0.8; and (3) tank pressure: 0.02 MPa; time: 24-36 h; the final thallus concentration is more than or equal to 2 hundred million/mL.
Example 2 Effect of Using the decomposing agent of the present invention
The using method comprises the following steps: and composting the organic materials layer by layer. 15-20 cm per layer, uniformly spreading the decomposition agent, urea or human and animal excreta of the invention example 1 layer by layer, and the dosage is as follows: 2 per mill of decomposition agent; 6-8 kg/ton urea or 100-200 kg/ton human and animal excrement and urine, and regulating water content to 60-70%.
The test effect is as follows:
the test was carried out in winter, at an ambient temperature of about 10 ℃. After rice is harvested, straws are smashed and then are piled for retting fermentation, one mu of rice straw is piled into two piles, and the size of the piles is 2 meters wide and 1.2 to 1.5 meters high. Spreading a layer of 20 cm thick with decomposing agent and human and animal feces layer by layer, adding water uniformly to control the water content of the material to be about 60-70%, and sealing the material with plastic film after the material is piled.
The use amount is as follows: the usage amount of the decomposing agent is 2 per mill of the material amount, namely 2kg of the decomposing agent is used for fermenting 1000 kg of materials.
As shown in Table 1, the decomposing inoculant provided by the invention can rapidly increase the temperature of compost materials, promote the growth and reproduction of normal-temperature decomposing bacteria, and degrade decomposed organic materials, and the same decomposition degree is 3-5 days earlier than that of similar products.
TABLE 1
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by the present specification, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (2)
1. A preparation method of a straw decomposition agent for decomposing at low temperature is characterized by comprising the following steps: respectively culturing Arthrobacter (Arthrobacter ariliati) YO-H, Bacillus subtilis, Saccharomyces cerevisiae, Aspergillus niger and Trichoderma harzianum until the bacterial concentration is 108Per mL, in a volume ratio of 1: 1: 1: 2: 2, uniformly mixing to obtain a mixed bacterial liquid, inoculating the obtained mixed bacterial liquid into wheat bran with the water content of 50% by mass according to the proportion of 1:10, culturing for 10 days, air-drying the mixture in a shade until the water content of the wheat bran is less than or equal to 25%, adding cellulase with the wheat bran mass content of 1% -1.5%, and uniformly mixing to obtain the straw decomposition agent, wherein the strain YO-H has the collection number of CGMCC NO. 4119;
the culture method of the strain YO-H is as follows:
1) activating strains: taking the lawn of the 1-ring strain YO-H from the bevel storage tube into 30mL of liquid culture medium, and culturing at 37 ℃ and 200-250 rpm for 20-24 hours; the preparation method of the liquid culture medium comprises the following steps: adding deionized water into bean sprouts, boiling for 20 minutes until the mass fraction of the bean sprouts is 25%, filtering to remove residues, supplementing the volume, adding 5% of glucose in mass fraction, dissolving and uniformly mixing, adjusting the pH value to 7.0, and sterilizing for 30 minutes at 121 ℃;
2) and (3) shake flask culture: sucking the bacterial liquid obtained in the step 1) by using a sterile pipettor into a shake flask, wherein the inoculation amount is as follows: 1 percent; temperature: 37 ℃; rotating speed: 200-250 rpm; culturing time: 20-24 h;
3) culturing in a fermentation tank: inoculating the shake flask cultured in the step 2) into a fermentation tank for amplification culture in an inoculation mode: flame inoculation; inoculation amount: 1 percent; temperature: 37 ℃; rotating speed: 180-200 rpm; the ventilation volume is 1: 0.8; and (3) tank pressure: 0.02 MPa; time: 24-36 h; obtaining bacterial liquid of strain YO-H.
2. The method for preparing straw decomposition agent for decomposition under low temperature according to claim 1, wherein the method for preparing liquid medium for shake flask in step 2) is as follows: adding deionized water into bean sprouts, boiling for 20 minutes until the mass fraction of the bean sprouts is 25%, filtering to remove residues, supplementing the volume, adding 5% of glucose in mass fraction, dissolving and uniformly mixing, adjusting the pH value to 7.0, and sterilizing for 30 minutes at 121 ℃.
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| CN201611204518.2A CN106701621B (en) | 2013-12-03 | 2013-12-03 | Preparation method of straw decomposition agent for decomposing at low temperature |
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| CN201611204518.2A CN106701621B (en) | 2013-12-03 | 2013-12-03 | Preparation method of straw decomposition agent for decomposing at low temperature |
| CN201310641595.4A CN103710282B (en) | 2013-12-03 | 2013-12-03 | Straw decomposing agent for quick start of decomposition at low temperature and its preparation method |
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| CN103966150B (en) * | 2014-06-03 | 2016-03-16 | 天津市农业资源与环境研究所 | A kind of agricultural crop straw decomposing microbial inoculum and using method thereof and effect |
| CN104193430A (en) * | 2014-08-27 | 2014-12-10 | 武汉瑞泽园生物环保科技有限公司 | Straw rotting agent and preparation method thereof |
| CN105746850A (en) * | 2014-12-16 | 2016-07-13 | 北京大北农科技集团股份有限公司 | Low-temperature fermentation feed raw material and application of same in preparation of mixed feed for pig |
| CN104788135B (en) * | 2015-04-07 | 2018-08-07 | 天津市农业资源与环境研究所 | Categorized consumer waste process for producing organoclay process system |
| CN105294185B (en) * | 2015-11-06 | 2018-12-04 | 东北农业大学 | The agricultural compost of low temperature that can be reduced agricultural non-point source pollution plays the preparation method of warm agent |
| CN105272424A (en) * | 2015-11-24 | 2016-01-27 | 东北农业大学 | Method for preparing low-temperature peasant family self-use decomposition maturing agent |
| CN105368744A (en) * | 2015-11-30 | 2016-03-02 | 甘肃明德伟业生物科技有限公司 | Enzyme-bacterium duplex straw decomposing agent and its preparation and application methods |
| CN106342908A (en) * | 2016-08-04 | 2017-01-25 | 王永强 | Compound microbial agent composition capable of reducing continuous cropping obstacles for crops |
| CN106244488A (en) * | 2016-08-25 | 2016-12-21 | 仇颖超 | A kind of preparation method of efficient straw decomposing agent |
| CN108504586B (en) * | 2017-12-06 | 2020-05-05 | 四川明湖环保科技有限公司 | Paenibacillus softening and straw decomposing agent |
| CN110835607B (en) * | 2018-08-17 | 2023-05-02 | 吉林农业大学 | Low-temperature efficient degradation straw composite bacteria containing saccharomyces cerevisiae and application thereof |
| CN109161495B (en) * | 2018-08-17 | 2020-04-28 | 吉林农业大学 | Composite microbial inoculum for efficiently degrading straw cellulose |
| CN110041118A (en) * | 2019-03-27 | 2019-07-23 | 广州国色生物科技有限公司 | A kind of efficient cryogenic straw decomposing inoculant |
| CN110041133A (en) * | 2019-05-07 | 2019-07-23 | 哈尔滨禾野农业有限公司 | A kind of bacteria agent and preparation method for straw decomposing |
| CN110551650B (en) * | 2019-08-09 | 2022-11-01 | 辽宁省农业科学院 | Bacillus subtilis for producing cellulase at low temperature and screening method thereof |
| CN110527644B (en) * | 2019-08-09 | 2022-11-01 | 辽宁省农业科学院 | Screening method and application of low-temperature straw composting composite microbial inoculum |
| CN111334456A (en) * | 2020-03-18 | 2020-06-26 | 吉林省农业科学院 | Preparation method and application of straw decomposition microbial inoculum used under low-temperature condition in north |
| CN113929509A (en) * | 2021-11-09 | 2022-01-14 | 吉林农业大学 | Decomposing inoculant for straw treatment and preparation method thereof |
| CN116333944B (en) * | 2023-05-11 | 2023-08-22 | 东北农业大学 | Arthrobacter C2 and application of recombinant Arthrobacter in lignin degradation |
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| CN102174398B (en) * | 2010-12-14 | 2012-10-31 | 北京沃土天地生物科技有限公司 | Composite microbiological bacterial agent used for returning maize straws to field and preparation method and applications thereof |
| CN102517217B (en) * | 2011-12-27 | 2013-09-25 | 宋保德 | Environment-friendly straw fermenting agent |
| CN103274771B (en) * | 2013-05-31 | 2015-11-25 | 上海创博生态工程有限公司 | A kind of straw decomposing inoculant and preparation method thereof |
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| CN103710282A (en) | 2014-04-09 |
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