CN102533553A - Organic matter decomposition combined fungicide for rapidly degrading rice straws and application method thereof - Google Patents
Organic matter decomposition combined fungicide for rapidly degrading rice straws and application method thereof Download PDFInfo
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Abstract
The invention discloses an organic matter decomposition combined fungicide for rapidly degrading rice straws and an application method thereof, which are applied to in-place returning-to-field high-efficiency stable fertilization of the straws. The fungicide contains Trichoderma koningii CGMCC NO.3.4290, Bacillus licheniformis CGMCC NO.4835 and bacillus subtilis CGMCC NO.1.1414 with the fungus content ratio being 1:1:1. The fungicide is obtained after mixing a product obtained after solid fermentation culture is performed on the Trichoderma koningii with a culture obtained after two kinds of bacillus liquid are cultured and a product adsorbed by wheat bran. The fineness of the fungicide does not exceed 0.18mm, and the water content is lower than 30 percent. As a spore serves as a main existence form, the fungicide disclosed by the invention has strong adaptability, is suitable for spraying and spreading, is easy for mechanized operations, is capable of greatly reducing labor strength, shortening life cycle and accelerating the decomposition speed of the straws and is beneficial to large-scale use.
Description
Technical field
The present invention relates to be applied to the organic material composting combination microbial inoculum and the application method thereof of going back the fertile a kind of quick degradation of rice straw stalk that uses of field efficient stable one-tenth on the spot of rice straw.
Background technology
Agricultural crop straw will produce nearly 200,000,000 tons of straw as a kind of renewable resources in the annual agricultural production process of China.Because natural decomposition needs the time longer, burning disposal is still in a large amount of uses, and the quick decomposition of straw also is used for and the field processing is the problem that receives much attention.So the agricultural crop straw decomposition utilizes for solving following energy dilemma and environmental problem significant.
The Hunan is a big agricultural province, and food crop are main with double cropping paddy rice.About 3,200 ten thousand tons of annual total stalk amount, wherein rice straw is about 2,755 ten thousand tons." the straw-returning lifting soil organism " project and the Hunan Province's rice straw that start on the occasion of country fully utilize the period that create the demonstration area; This institute intends and develops the strong and high organic material composting agent of degradation efficiency of adaptive faculty to south ecology, envrionment conditions and cropping system.The application of this technology and products thereof can be accelerated the speed of becoming thoroughly decomposed of stalk, improves the content of soil with organic matter, available potassium etc., can quicken the rice nutrition supply, promote effective tillering, thereby increase plant height and effective fringe, promotes rice yield.Thereby improved the stalk comprehensive utilization ratio effectively, can obtain favorable economic benefit, ecological benefits and social benefit.
Summary of the invention
The object of the present invention is to provide a kind of be suitable for the stalk field on the spot also the field condition, rapidly and efficiently the decomposition stalk, have the organic material composting combination microbial inoculum and the application method thereof of a kind of quick degradation of rice straw stalk of certain stability.
The technical scheme that the present invention adopted
Contain the mould CGMCC NO.3.4290 of 1: 1: 1 healthy and free from worry wood of bacteria containing amount proportioning in a kind of organic material composting combination microbial inoculum of quick degradation of rice straw stalk; Bacillus licheniformis (Bacillus licheniformis) CGMCC NO.4835; Subtilis CGMCC NO.1.1414; Described microbial inoculum is mixed by following three kinds of compositions and forms: the product of product, the culture after the subtilis CGMCC NO.1.1414 liquid fermentation and culture and the wheat bran absorption of product, the culture after the Bacillus licheniformis CGMCC NO.4835 liquid fermentation and culture and wheat bran absorption after the mould CGMCC NO.3.4290 of healthy and free from worry wood cultivates through liquid fermenting, solid fermentation successively.
The mould CGMCC NO.3.4290 of described healthy and free from worry wood cultivates 45-48h in 25-28 ℃ of solid fermentation, and bacterial content is not less than 10 in the product
8Individual/g, described subtilis is cultivated 22-24h at 34-37 ℃, and described Bacillus licheniformis is behind 45-55 ℃ of cultivation 36-40h, and with culture and wheat bran absorption, bacterial content all is not less than 10 in the product respectively
9Individual/g, described bacterial content comprises thalline and the total content of gemma.
Bacterial content is not less than 2 * 10 in the described microbial inoculum
8Individual/g, the microbial inoculum fineness is no more than 0.18mm, and water cut is not higher than 30%.4. microbial inoculum according to claim 1 is characterized in that, the mould CGMCC NO.3.4290 of described healthy and free from worry wood solid fermentation detailed process is following:
After at first the mould CGMCC NO.3.4290 of described healthy and free from worry wood being cultivated through inclined-plane seed activation, shake-flask seed, the shake-flask seed nutrient solution is inoculated in the liquid fermentation tank substratum cultivates incubation time 22-24h, temperature 25-28 ℃, rotating speed 120-150rpm; Air flow 0.4-0.5VVM; Inoculum size accounts for the 10-15% of culture volume, and the substratum loading amount accounts for the 60-70% of fermentor tank volume in the fermentor tank, and the liquid fermentation medium compound method is: sucrose 20g, yeast powder 0.2g, malt meal 3.0g, NaCl 0.1g, CaCl
22H
2O 0.1g, KH
2PO
41.0g, MgSO
47H
2O 0.5g, (NH
4)
2SO
40.5g adding distil water is to 1L, pH6.5,121 ℃, 0.1MPa autoclaving 30min;
Inoculation accounts for the fermentation culture seed liquor of solid medium weight 10-15% then, and 25-28 ℃ of solid fermentation cultivated 45-48h; Solid fermentation culture medium preparation method is: Semen Maydis powder 10g, wheat bran 90g, (NH
4)
2SO
41.0g, KH
2PO
40.2g, MgSO
47H
2O 0.05g, 80mL zero(ppm) water, 121 ℃, 0.1MPa autoclaving 30min, the loading amount of substratum is the 60-70% of volume in the solid-state fermentation tank.
After described Bacillus licheniformis CGMCC NO.4835 cultivates through inclined-plane seed activation, shake-flask seed; The shake-flask seed nutrient solution is inoculated in the liquid fermentation tank substratum cultivates; The compound method of used liquid fermentation medium is: starch 5.0g, dextrin 20g, soybean cake powder 25g; Yeast powder 3.0g, CaCO
312g, (NH
4)
2SO
410g, NaCl 4.0g, KH
2PO
40.2g adding distil water is to 1L, pH7.2-7.4,121 ℃, 0.1MPa autoclaving 30min; Incubation time 36-40h, temperature 45-55 ℃, rotating speed 180-200rpm, air flow 0.8-1.0VVM, inoculum size accounts for the 8-10% of culture volume, and the liquid amount of substratum is the 55-65% of fermentor tank volume in the fermentor tank.
After described subtilis CGMCC NO.1.1414 cultivates through inclined-plane seed activation, shake-flask seed; The shake-flask seed nutrient solution is inoculated in the liquid fermentation tank substratum cultivates, the compound method of used liquid fermentation medium is: glucose 5.0g, sucrose 3.0g; Soybean cake powder 15g; Yeast powder 10g, peptone 5.0g, CaCO
31.0g, FeCl
36H
2O 0.1g, KH
2PO
44.0g, MgSO
47H
2O 0.5g, MnSO
4H
2O 0.062g, adding distil water be to 1L, pH7.2,121 ℃, 0.1MPa autoclaving 30min; Incubation time 22-24h, temperature 34-37 ℃, rotating speed 180-200rpm, air flow 0.8-1.0VVM, inoculum size accounts for the 8-10% of culture volume, and the liquid amount of substratum is the 55-65% of fermentor tank volume in the fermentor tank.
In the above-mentioned fungicide preparation process, during wheat bran absorption, its part by weight accounts for 70-80%.
Described microbial inoculum is used to improve paddy rice proterties and output; Specifically be to add in the rice straw, be used to quicken the content that rice straw becomes thoroughly decomposed, improves soil with organic matter, available potassium, quicken the rice nutrition supply, promote effective tillering, thereby increase plant height and effective fringe, promote rice yield.
The principle and the beneficial effect of combination bacterial strain of the present invention are: the composition that the present invention is directed to stalk; With multiple lignoenzyme vigor, cellulase activity, semicellulose enzyme activity and straw degradative rate serves as to detect index, chooses the mould CGMCC NO.3.4290 of healthy and free from worry wood especially; Bacillus licheniformis CGMCC NO.4835; Subtilis CGMCC NO.1.1414.Again the product of three kinds of strain culturing is mixed with the microbial inoculum that forms and is applied to straw also field field plot trial on the spot.Because after the stalk structure is studied; It is found that xylogen combines with the covalent linkage form with semicellulose; Cellulosic molecule is embedded in wherein, forms a kind of natural cover for defense, make cellulase etc. be difficult for contacting with cellulosic molecule; And the complicacy of water-insoluble, the chemical structure of xylogen has caused the refractory organics of stalk.Therefore want thorough degraded cellulose, at first must solve the degradation problem of xylogen.Utilize at first that healthy and free from worry wood is mould to decompose the composition wax and xylogen is preferentially degraded to stalk is outer difficult; Actively construct the strain combinations that improves straw class stalk decomposition efficient through cellulose-binding degradation bacteria (Bacillus licheniformis and subtilis) again.
Be made up of xylogen, Mierocrystalline cellulose, semicellulose etc. to stalk, the present invention will secrete that the healthy and free from worry wood of highly active lignoenzyme, cellulase and hemicellulase is mould, Bacillus licheniformis and bacillus subtilis strain make up.Because the combination of this bacterial classification has multiple bacterial strain, has certain stability, and all bacterium all exist with the gemma form, and growth conditions is suitable for wide, is prone to the survival breeding, especially on utilization of carbon source, has very strong synergetic property, therefore are easy to improve the stalk efficient of becoming thoroughly decomposed.This straw degradative microbial inoculum is combined in 14 days of degrading straw, and it is stable that the pH value keeps, and lignoenzyme, cellulase activity and living bacteria count amount remain at 10
7Individual/more than the g, the straw degradative rate constantly increases, and explains that this straw degradative microbial inoculum has very strong the adaptation and regulating power to pH, can maintain property stablize, and in the function performance, also has certain stability simultaneously.This microbial inoculum is main existence form with spore, and fineness is in the 0.18mm levels, and flexibility is strong; In being applied to the returning field into fertilizer on the spot of rice straw, be suitable for spreading fertilizer over the fields, spraying, be easy to mechanized operation; Can reduce labour intensity greatly; Shorten life cycle, quicken stalk and become thoroughly decomposed, help extensive use.
Bacillus licheniformis (Bacillus licheniformis) preserving number is in the bacterial classification combination of the present invention: CGMCC NO.4835; It is bacterial strain by the applicant oneself screening; And carried out preservation at the China Committee for Culture Collection of Microorganisms common micro-organisms center of the Institute of Microorganism, Academia Sinica that is positioned at No. 3, No. 1 institute in North Star West Road, Chaoyang District, BeiJing, China city on May 9th, 2011; And applied for that patent, application number are 201110166962.0.
The mould CGMCC NO.3.4290 of subtilis CGMCC NO.11414 and healthy and free from worry wood all buys the institute of microbiology in the Chinese Academy of Sciences.
Embodiment
Be intended to further specify the present invention below in conjunction with embodiment, and unrestricted the present invention.
Embodiment 1 fungicide preparation of the present invention
1) the mould CGMCC NO.3.4290 of healthy and free from worry wood is cultivated the back to the liquid fermentation tank fermentation culture through inclined-plane seed, shake-flask seed; Finally in solid-state fermentation tank, cultivate and produce spore, seed culture used culture medium preparation method in inclined-plane is: peptone 6.0g, potato 200g; NaCl 2.0g, MgSO
41.0g, sucrose 20g, KH
2PO
41.0g agar 18-20g, adding distil water are to 1L, pH is natural, 121 ℃, and 0.1MPa autoclaving 30min.Inclined-plane seed culture time 30-36h, temperature 25-28 ℃.Inclined-plane seed inoculation two to three encircles to shake-flask seed and cultivates, and shakes at the 500mL triangle that the substratum loading amount is 100mL in the bottle.The used culture medium preparation method of shake-flask seed is: soybean cake powder 1.5g, sucrose 20g, MgSO
41.5g, KH
2PO
43.0g adding distil water is to 1L, pH is natural, 121 ℃, and 0.1MPa autoclaving 30min; Incubation time 22-24h, temperature 25-28 ℃, rotating speed 120-150rpm cultivates in the liquid fermentation tank substratum; Incubation time 22h, 28 ℃ of temperature, rotating speed 150rpm, air flow 0.5VVM.The liquid fermentation medium compound method is: sucrose 20g, yeast powder 0.2g, malt meal 3.0g, NaCl 0.1g, CaCl
22H
2O 0.1g, KH
2PO
41.0g, MgSO
47H
2O 0.5g, (NH
4)
2SO
40.5g adding distil water is to 1L, pH6.5,121 ℃, 0.1MPa autoclaving 30min; Inoculum size accounts for 15% of culture volume, and the substratum loading amount accounts for 70% of volume in the fermentor tank.
2) inoculate the liquid fermentation and culture seed liquor that accounts for solid medium weight 15% then, 25 ℃ of solid fermentations are cultivated 48h; The compound method of solid medium is: Semen Maydis powder 10g, wheat bran 90g, (NH
4)
2SO
41.0g, KH
2PO
40.2g, MgSO
47H
2O0.05g, 80mL zero(ppm) water, 121 ℃, 0.1MPa autoclaving 30min.The loading amount of substratum is 70% of a volume in the solid-state fermentation tank.Bacterial content is in 3-6 * 10 in the product
8Individual/g, bacterial content comprises thalline and the total content of gemma.
3) Bacillus licheniformis CGMCC NO.4835 cultivates the back to fermentation culture through inclined-plane seed, shake-flask seed earlier; Wherein seed culture used culture medium preparation method in Bacillus licheniformis CGMCC NO.4835 inclined-plane is: glucose 4.0g; Yeast extract paste 4.0g, malt meal 10.0g, CaCO
32.0g agar 20g, adding distil water be to 1L, pH7.2, incubation time 24-26h, temperature 45-55 ℃.Inclined-plane seed inoculation one encircles to shake-flask seed and cultivates, and shakes at the 500mL triangle that the substratum loading amount is 100mL in the bottle.Used culture medium preparation method is: starch 30.0g, soybean cake powder 5.0g, yeast extract paste 5.0g, CaCO
35.0g, (NH
4)
2SO
44.0g, NaCl 5.0g, KH
2PO
40.12g adding distil water is to 1L, pH7.2,121 ℃, 0.1MPa autoclaving 30min.Incubation time 24-30h, temperature 45-55 ℃, rotating speed 200-220rpm.Inoculate the seed culture fluid that accounts for fermention medium volume 8% then and in fermention medium, cultivate 36h, 55 ℃ of temperature, rotating speed 200rpm, air flow 0.8VVM.The liquid amount of substratum is 65% in the fermentor tank.The compound method of used fermention medium is: starch 5.0g, dextrin 20g, soybean cake powder 25g, yeast powder 3.0g, CaCO
312g, (NH
4)
2SO
410g, NaCl 4.0g, KH
2PO
40.2g adding distil water is to 1L, pH7.2,121 ℃, 0.1MPa autoclaving 30min.
With the culture and the wheat bran absorption of fermentation culture, bacterial content is all in 4-6 * 10 in the product
9Individual/g, bacterial content comprises thalline and the total content of gemma.
4) subtilis CGMCC NO.1.1414 cultivates the back to fermentation culture through inclined-plane seed culture, shake-flask seed earlier; The compound method of used inclined-plane seed culture medium is: Carnis Bovis seu Bubali cream 3.0 peptones 10.0 sodium-chlor 5.0; Adding distil water is to 1L; PH7.2,121 ℃, 0.1MPa autoclaving 30min; Incubation time 22-24h, temperature 34-37 ℃.Inclined-plane seed inoculation one encircles to shake-flask seed and cultivates, and shakes at the 500mL triangle that the substratum loading amount is 100mL in the bottle.Shake-flask seed is cultivated used culture medium preparation method: Carnis Bovis seu Bubali cream 5.0 yeast extracts 5.0 peptones 10.0 glucose 5.0 sodium-chlor 5.0 zero(ppm) water 1000mlpH7.0,121 ℃, 0.1MPa autoclaving 30min.Incubation time 22-24h, temperature 34-37 ℃, rotating speed 200-220rpm.The shake-flask seed nutrient solution of inoculation 8% is to fermention medium, and the liquid amount of substratum is 65% in the fermentor tank.The compound method of used fermention medium is: glucose 5.0g, sucrose 3.0g, soybean cake powder 15g, yeast powder 10g, peptone 5.0g, CaCO
31.0g, FeCl
36H
2O 0.1g, KH
2PO
44.0g, MgSO
47H
2O 0.5g, MnSO
4H
2O 0.062g, adding distil water be to 1L, pH7.2,121 ℃, 0.1MPa autoclaving 30min.Incubation time 22h, 37 ℃ of temperature, rotating speed 200rpm, air flow 0.8VVM.With the culture and the wheat bran absorption of fermentation culture, bacterial content is all in 6-8 * 10 in the product
9Individual/g, bacterial content comprises thalline and the total content of gemma.
5) product after process liquid fermenting, solid fermentation are cultivated successively with the mould CGMCC NO.3.4290 of above-mentioned healthy and free from worry wood; With the microbial inoculum that obtains after the product of culture after the Bacillus licheniformis CGMCC NO.4835 liquid fermentation and culture and wheat bran absorption, the product of culture and wheat bran absorption after the subtilis CGMCC NO.1.1414 liquid fermentation and culture mix.The part by weight of wheat bran absorption is 70%, the mould CGMCC NO.3.4290 of 1: 1: 1 healthy and free from worry wood of bacteria containing amount proportioning; Bacillus licheniformis CGMCC NO.4835; Subtilis CGMCC NO.1.1414; Bacterial content is 2 * 10 in the microbial inoculum
8Individual/g; The microbial inoculum fineness is 0.18mm, and water cut is not higher than 30%.
Embodiment 2
Bacterial preparation process is with embodiment 1, be incubation time when cultivating in the mould CGMCC NO.3.4290 of the healthy and free from worry wood liquid fermentation tank substratum be 24h, 25 ℃ of temperature, rotating speed 120rpm, air flow 0.4VVM.Inoculum size accounts for 10% of culture volume, and the substratum loading amount is 60% in the fermentor tank.Inoculation accounts for the liquid culture seed liquor of solid medium weight 10% then, and 28 ℃ of solid fermentations are cultivated 45h; The loading amount of substratum is 60% of a volume in the solid-state fermentation tank.Bacterial content is in 3-6 * 10 in the product
8Individual/g, bacterial content comprises thalline and the total content of gemma.
Bacillus licheniformis CGMCC NO.4835, inoculation accounts for the seed culture fluid of fermention medium volume 10% and in fermention medium, cultivates 40h, 45 ℃ of temperature, rotating speed 180rpm, air flow 1.0VVM.The liquid amount of substratum is 55% in the fermentor tank.With culture and wheat bran absorption, bacterial content is all in 4-6 * 10 in the product
9Individual/g, bacterial content comprises thalline and the total content of gemma.
Subtilis CGMCC NO.1.1414, inoculation 10% shake-flask seed nutrient solution to fermention medium, incubation time 24h, 35 ℃ of temperature, rotating speed 180rpm, air flow 1.0VVM.The liquid amount of substratum is 55% in the fermentor tank.With the culture and the wheat bran absorption of fermentation culture, bacterial content is all in 6-8 * 10 in the product
9Individual/g, bacterial content comprises thalline and the total content of gemma.
The part by weight of wheat bran absorption is 80%, and bacterial content is 2 * 10 in the microbial inoculum that obtains at last
8Individual/g, the microbial inoculum fineness is 0.18mm, and water cut is not higher than 30%.
The contrast of embodiment 3 microbial inoculum straw degradative rates
The cultural method that the rice straw degradation rate is measured substratum is: fresh rice straw is cut into the long short stalk of 1cm; Take by weighing the triangular flask of 20g adding 500mL, every bottle of following microbial inoculum and 100mL zero(ppm) water that adds 50g again mixes; Nature pH value is in 121 ℃, the following sterilization of 0.1MPa 30min.
At first cellulose-degrading bacteria (being two kinds of genus bacillus of the present invention) being cultivated behind the 24h wheat bran absorption respectively at 34 ℃, to process concentration be 10
8The spore pulvis of individual/g mixes according to 1: 1 volume ratio, and the straw degradative rate is 39.6%, is higher than single strain; And the present invention is composite with two kinds of cellulose-degrading bacterias and lignin-degrading bacteria, and the straw degradative rate brings up to 50.2%.Explain that the bacterial strain in this combination exists certain synergetic property, does not have antagonism property between them.
Bacterial classification combination microbial inoculum of the present invention is an index with the rice straw degradation rate, and its degradation rate to rice straw is significantly improved than single bacterial classification, has reached 50.2%.
The application of embodiment 4 microbial inoculums of the present invention in the shortening rice straw becomes thoroughly decomposed the time
5 processing are established in test, 3 repetitions.Handle 1: conventional fertilizer application+straw incorporation+close edge board board organic material composting agent; Handle 2: conventional fertilizer application+straw incorporation+microbial inoculum of the present invention; Handle 3: conventional fertilizer application+straw incorporation+paddy continuous heavy rain board organic material composting agent; Handle 4: conventional fertilizer application+straw incorporation; Handle 5: conventional fertilizer application; Late rice test site random alignment, sub-district area 33.3M
2Result of the test shortens and becomes thoroughly decomposed the time than handling 4 for handling 2, handle 1 with handle 3 and be excellent (seeing Table 1).
The table 1 different treatment straw degree record table that becomes thoroughly decomposed
Annotate: under the general condition; Stalk decomposition degree is qualitative comparison; Be decided to be 1,2,3,4 grade to Huang, little Huang, brown Huang, black Huang in the stalk color respectively during statistics; Musty in the stalk smell, ammonia flavor, vinosity, putrid taste are decided to be 1,2,3,4 grade respectively, hard, the Microsoft in the feel softening degree, soft, rot to be decided to be 1,2,3,4 grade respectively, the decomposition degree numerical value during each is handled during statistics is three big index value sums; The stalk of big more this processing of expression of numerical value rots fast more, and the effect of becoming thoroughly decomposed is obvious more.
The application of embodiment 5 microbial inoculums of the present invention in the paddy rice economic characters are improved
With case study on implementation 4 same treatment group.Processing 2 ratio processing 1 and processing 3 every mu of average plant heights and effective fringe all have increase, and rice yield also has increase, and while and processing 4 and processing 5 significances of difference have economic benefit (seeing table 2,3,4,5,6).
Table 2 different treatment economic characters table (unit: cm, ten thousand/667m
2, %, g, kg/667m
2)
Table 3 different treatment rate schedule (unit: strain/667m
2, kg/667m
2)
Table 4 variance analysis
| Source of variation | Degree of freedom | Sum of squares | All square | The F value | F0.05 | F0.01 |
| Between processing | 4 | 19.871 | 4.968 | 114.638 | 3.478 | 5.994 |
| Error | 10 | 0.433 | 0.043 | - | - | - |
| Total variation | 14 | 20.304 | - | - | - | - |
[0049]Table 5 multiple comparisons (LSD method)
Table 6 different treatment performance analysis (unit: unit/667m
2, kg/667m
2)
Embodiment 6 microbial inoculum working conditions of the present invention and method of use
1. working conditions: the field optimum inoculation amount of this straw degradative combination microbial inoculum is 4 ‰, and promptly every kilogram of rice straw need be inoculated the 4g degradation bacterial agent; When the land for growing field crops applies microbial inoculum, need apply nitrogen and phosphorus.The applied amount of every kilogram of rice straw nitrogen is 12g, and the additional amount of phosphorus is 5g, generally helps the rice straw degraded more greatly more, therefore can confirm the phosphorus magnitude of recruitment based on actual conditions.
2. method of use: combine rice straw also the field time, spread fertilizer over the fields or spray microbial inoculum of the present invention.Consumption: every mu of 2-3 kilogram, spread fertilizer over the fields on rice straw or spray microbial inoculum of the present invention and get final product.Apply nitrogenous fertilizer and phosphate fertilizer simultaneously, then rice straw is ploughed under underground, also applicable to the rice straw fermentation of becoming thoroughly decomposed under the condition of banking up.
Claims (9)
1. the organic material composting of quick degradation of rice straw stalk combination microbial inoculum is characterized in that, contains the mould CGMCC NO.3.4290 of 1: 1: 1 healthy and free from worry wood of bacteria containing amount proportioning in the described microbial inoculum; Bacillus licheniformis CGMCC NO.4835; Subtilis CGMCC NO.1.1414; Described microbial inoculum is mixed by following three kinds of compositions and forms: the product of product, the culture after the subtilis CGMCC NO.1.1414 liquid fermentation and culture and the wheat bran absorption of product, the culture after the Bacillus licheniformis CGMCC NO.4835 liquid fermentation and culture and wheat bran absorption after the mould CGMCC NO.3.4290 of healthy and free from worry wood cultivates through liquid fermenting, solid fermentation successively.
2. microbial inoculum according to claim 1 is characterized in that, the mould CGMCC NO.3.4290 of described healthy and free from worry wood cultivates 45-48h in 25-28 ℃ of solid fermentation, and bacterial content is not less than 10 in the product
8Individual/g, described subtilis is cultivated 22-24h at 34-37 ℃, and described Bacillus licheniformis is behind 45-55 ℃ of cultivation 36-40h, and with culture and wheat bran absorption, bacterial content all is not less than 10 in the product respectively
9Individual/g, described bacterial content comprises thalline and the total content of gemma.
3. microbial inoculum according to claim 1 is characterized in that bacterial content is not less than 2 * 10 in the described microbial inoculum
8Individual/g, the microbial inoculum fineness is no more than 0.18mm, and water cut is not higher than 30%.
4. microbial inoculum according to claim 1 is characterized in that, the mould CGMCC NO.3.4290 of described healthy and free from worry wood solid fermentation detailed process is following:
After at first the mould CGMCC NO.3.4290 of described healthy and free from worry wood being cultivated through inclined-plane seed activation, shake-flask seed, the shake-flask seed nutrient solution is inoculated in the liquid fermentation tank substratum cultivates incubation time 22-24h, temperature 25-28 ℃, rotating speed 120-150rpm; Air flow 0.4-0.5VVM; Inoculum size accounts for the 10-15% of culture volume, and the substratum loading amount accounts for the 60-70% of fermentor tank volume in the fermentor tank, and the liquid fermentation medium compound method is: sucrose 20g, yeast powder 0.2g, malt meal 3.0g, NaCl 0.1g, CaCl
22H
2O 0.1g, KH
2PO
41.0g, MgSO
47H
2O 0.5g, (NH
4)
2SO
40.5g adding distil water is to 1L, pH6.5,121 ℃, 0.1MPa autoclaving 30min;
Inoculation accounts for the fermentation culture seed liquor of solid medium weight 10-15% then, and 25-28 ℃ of solid fermentation cultivated 45-48h; Solid fermentation culture medium preparation method is: Semen Maydis powder 10g, wheat bran 90g, (NH
4)
2SO
41.0g, KH
2PO
40.2g, MgSO
47H
2O 0.05g, 80mL zero(ppm) water, 121 ℃, 0.1MPa autoclaving 30min, the loading amount of substratum is the 60-70% of volume in the solid-state fermentation tank.
5. microbial inoculum according to claim 1 is characterized in that, after described Bacillus licheniformis CGMCC NO.4835 cultivates through inclined-plane seed activation, shake-flask seed; The shake-flask seed nutrient solution is inoculated in the liquid fermentation tank substratum cultivates; The compound method of used liquid fermentation medium is: starch 5.0g, dextrin 20g, soybean cake powder 25g; Yeast powder 3.0g, CaCO
312g, (NH
4)
2SO
410g, NaCl 4.0g, KH
2PO
40.2g adding distil water is to 1L, pH7.2-7.4,121 ℃, 0.1MPa autoclaving 30min; Incubation time 36-40h, temperature 45-55 ℃, rotating speed 180-200rpm, air flow 0.8-1.0VVM, inoculum size accounts for the 8-10% of culture volume, and the liquid amount of substratum is the 55-65% of fermentor tank volume in the fermentor tank.
6. microbial inoculum according to claim 1 is characterized in that, after described subtilis CGMCC NO.1.1414 cultivates through inclined-plane seed activation, shake-flask seed; The shake-flask seed nutrient solution is inoculated in the liquid fermentation tank substratum cultivates, the compound method of used liquid fermentation medium is: glucose 5.0g, sucrose 3.0g; Soybean cake powder 15g; Yeast powder 10g, peptone 5.0g, CaCO
31.0g, FeCl
36H
2O 0.1g, KH
2PO
44.0g, MgSO
47H
2O 0.5g, MnSO
4H
2O 0.062g, adding distil water be to 1L, pH7.2,121 ℃, 0.1MPa autoclaving 30min; Incubation time 22-24h, temperature 34-37 ℃, rotating speed 180-200rpm, air flow 0.8-1.0VVM, inoculum size accounts for the 8-10% of culture volume, and the liquid amount of substratum is the 55-65% of fermentor tank volume in the fermentor tank.
7. microbial inoculum according to claim 1 is characterized in that, during wheat bran absorption, its part by weight accounts for 70-80%.
8. the application method of any described microbial inoculum of claim 1-7 is characterized in that, is used to improve paddy rice proterties and output.
9. application method according to claim 8 is characterized in that, adds in the rice straw; Be used to quicken the content that rice straw becomes thoroughly decomposed, improves soil with organic matter, available potassium; Quicken the rice nutrition supply, promote effective tillering, thereby increase plant height and effective fringe, promote rice yield.
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