CN101838613A - Agricultural straw fast-degradation decomposition agent strain composition - Google Patents

Agricultural straw fast-degradation decomposition agent strain composition Download PDF

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Publication number
CN101838613A
CN101838613A CN200910172426A CN200910172426A CN101838613A CN 101838613 A CN101838613 A CN 101838613A CN 200910172426 A CN200910172426 A CN 200910172426A CN 200910172426 A CN200910172426 A CN 200910172426A CN 101838613 A CN101838613 A CN 101838613A
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China
Prior art keywords
decomposition agent
hlg
strain
combination
hrqj
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CN200910172426A
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Inventor
郭夏丽
李海泉
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HENAN HENGLONGTAI BIO-ENGINEERING CO LTD
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HENAN HENGLONGTAI BIO-ENGINEERING CO LTD
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Publication of CN101838613A publication Critical patent/CN101838613A/en
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Abstract

The invention discloses an agricultural straw fast-degradation decomposition agent strain composition, which is applied to the in-place returning-to-farmland and highly-efficient and stable fertilizer-formation of agricultural mechanical straws. The decomposition agent strain composition is HLG+X2+L3+HRQJ, namely, the decomposition agent strain composition is prepared by compositing four fungus strains which are HLG Penicillium oxalicum, X2 cellulose degradation fungi, L3 lignin degradation fungi and HRQJ abnormal Pichia anomala respectively in the ratio of 1 to 1 to 1 to 1, culturing the fungus strains at 30 DEG C for 3 days respectively, preparing spore suspension and bacterial suspension with the bacteria content of 108 per mL, taking 10 mL of spore suspension and 10 mL of bacterial suspension and inoculating the spore suspension and the bacterial suspension to a microbial inoculum preparation culture medium, wherein inoculum concentration is 4 percent. Due to spores serving as a primary existence form, the microbial inoculum of the invention has the characteristics of high adaptability, application to spraying and easy mechanical operation, can greatly reduce labor intensity, shorten a life cycle and speed up the decomposition of the straws, and is favorable for large-scale use.

Description

A kind of decomposition agent strain combination of agricultural straw fast-degradation
Technical field: the present invention relates to be applied to the also decomposition agent strain combination of the fertile a kind of agricultural straw fast-degradation that uses of field efficient stable one-tenth on the spot of agricultural machine stalk.
Background technology: the stalk agent of becoming thoroughly decomposed is remarkable day by day in the effect and the effect that promote the soil organism, make rational use of resources, solve aspects such as environmental pollution.Present most of stalk agent product that becomes thoroughly decomposed is to research and develop the microbial inoculum product of production at becoming thoroughly decomposed feces of livestock and poultry or itself and stalk mixture under the condition of banking up, at stalk on the spot also the field research and development become thoroughly decomposed agent but seldom.In " the straw-returning lifting soil organism " project that country promotes, stalk on the spot also the field agent of becoming thoroughly decomposed of using also exist less stable, problem such as the bacterial classification of use and bacterial classification combination/collocation are unreasonable.How to research and develop the stalk that is applicable to " the straw-returning lifting soil organism " project agent product that becomes thoroughly decomposed, filter out excellent property, have the straw degradative bacterium of very strong decay, and they are carried out the organic assembling compatibility, the stalk decomposition microbial inoculum of producing efficient stable is the work of the task of top priority.
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 decomposition agent strain combination of a kind of agricultural straw fast-degradation of certain stability.
The technical solution adopted in the present invention is: decomposition agent strain is combined as: HLG+X2+L3+HRQJ; Promptly by HLG penicillium oxalicum (Latin formal name used at school: Penicillium oxalicum; Unit is identified in check: Institute of Microorganism, Academia Sinica; The date is identified in check: on August 25th, 2009; No. the 167th, (2009) little searching), X2 cellulose-degrading bacteria, L3 lignin-degrading bacteria and unusual complete red female bacterium (the Latin formal name used at school: Pichia anomala of HRQJ numbering is identified in check:; Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center; Preservation date: on December 18th, 2008; Depositary institution is called for short and numbering: CGMCCNo:2814) four kinds of fungi strains are 1: 1: 1 by proportioning: 1 compound composition; Its preparation technology cultivates HLG, X2, four kinds of bacterium of L3, HRQJ 3 days in 30 ℃ separately, makes 10 8The spore suspension of individual/mL and bacteria suspension are respectively got 10mL and are inoculated in the fungicide preparation substratum, and inoculum size is 4%, and in 30 ℃ of cultivations, incubation time is 2-3 days, and bacterial content can reach 10 8Individual/mL.
The principle and the beneficial effect of combination of the present invention are: the composition that the present invention is directed to stalk, with multiple cellulase activity, hemicellulase work, lignoenzyme work and straw degradative rate serves as to detect index, with the stalk is substratum, filters out X2 cellulose-degrading bacteria and L3 lignin-degrading bacteria from occurring in nature.Suitable pH is about 5 because three plain enzymes such as cellulase, hemicellulase, lignoenzyme are lived, in addition, in order to remove the feedback inhibition of cellulase activity and hemicellulase end product alive, filtering out the degraded end product that can utilize Mierocrystalline cellulose, hemicellulose from nature is the yeast HRQJ that glucose, wood sugar produce acid.
Form by Mierocrystalline cellulose, hemicellulose, xylogen etc. at stalk, HLG, X2, L3 bacterial strain that the present invention will secrete highly active cellulase, hemicellulase and lignoenzyme make up, but because the inhibition of cellulase and the hemicellulase end product of often being degraded alive, and yeast can utilize glucose, these end products of wood sugar to grow and ferment, can remove this restraining effect, yeast also has the acid of producing, the beneficial effect of giving birth to the factor of secretion simultaneously, therefore yeast is combined, can promote the degraded of stalk.Because the combination of this bacterial classification has multiple bacterial strain, has certain stability, and all bacterium be fungi all, and growth conditions is similar, especially has very strong synergy on utilization of carbon source, so is easy to mixed culture, can reduce the production cost of microbial inoculum.This straw degradative bacterium is combined in 14 days of degrading straw, it is stable that the pH value keeps, cellulose degradation enzymic activity and fungi quantity remain higher, the straw degradative rate constantly increases, illustrate that this straw degradative microbial inoculum has stronger regulating power to pH, can maintain property stablize, in the function performance, also have certain stability simultaneously.This microbial inoculum is main existence form with spore, and adaptability is strong, in being applied to the returning field into fertilizer on the spot of agricultural stalk, is suitable for spraying, and is easy to mechanized operation, can reduce labour intensity greatly, shortens life cycle, quickens stalk and becomes thoroughly decomposed, and helps extensive use.
HLG in the bacterial classification combination of the present invention and the biological characteristics of HRQJ and bacterial classification check are identified and the preservation brief description:
1, HLG penicillium oxalicum: Latin formal name used at school: Penicillium oxalicum;
Unit is identified in check: Institute of Microorganism, Academia Sinica;
Unit address is identified in check: the Datun Road, Chaoyang District, Beijing City;
Numbering is identified in check: No. the 167th, (2009) little searching
The date is identified in check: on August 25th, 2009;
The HLG penicillium oxalicum belongs to the Deuteromycotina fungi.Bacterium colony is smooth, greyish-green; The bacterium colony back side is light brown.The conidiophore wall is smooth, and is colourless, and most two-wheels are given birth to, and the stigma size is (140～200) * (3.5～4) microns.The conidium ellipse, smooth, size is (4.5～5.3) * (2.5～3) microns, conidium produces in a large number.
2, the unusual Pichia yeast of HRQJ: Latin formal name used at school: Pichia anomala;
Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center;
Depositary institution address: Datun Road, Chaoyang District, Beijing City;
Preservation date: on December 18th, 2008;
Depositary institution is called for short and numbering: CGMCC No:2814)
The unusual Pichia yeast of HRQJ is cell sphere, ellipse, sausage shape, and size is (2.4～4.8) * (2.4～6) microns, and bacterium colony shows smooth, is the cheese shape, oyster white.Energy glucose fermentation, assimilation wood sugar, growth temperature is 30～37 ℃.
Embodiment: embodiment 1: this decomposition agent strain is combined as: the HLG+X2+L decomposition agent strain is combined as: HLG+X2+L3+HRQJ; Promptly by HLG penicillium oxalicum (Latin formal name used at school: Penicillium oxalicum; Unit is identified in check: Institute of Microorganism, Academia Sinica; Identify the date: on August 25th, 2009; No. the 167th, (2009) little searching), X2 cellulose-degrading bacteria, L3 lignin-degrading bacteria and unusual complete red female bacterium (the Latin formal name used at school: Pichia anomala of HRQJ identify numbering:; Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center; Preservation date: on December 18th, 2008; Depositary institution is called for short and numbering: CGMCC No:2814) four kinds of fungi strains are 1: 1: 1 by proportioning: 1 compound composition; Its preparation technology cultivates HLG, X2, four kinds of bacterium of L3, HRQJ 3 days in 30 ℃ separately, makes 10 8The spore suspension of individual/mL and bacteria suspension are respectively got 10mL and are inoculated in the fungicide preparation substratum, and inoculum size is 4%, and in 30 ℃ of cultivations, incubation time is 2-3 days, and bacterial content can reach 10 8Individual/mL.HLG, X2, four kinds of bacterial contents of L3, HRQJ are 10 in its finished product 6～10 10Individual/mL, HLG, X2, L3 bacterial strain mainly exist with spore form in the microbial inoculum.
Embodiment 2: culture medium culturing:
The cultural method of HLG, X2, the used substratum of L3 single strain is: K 2HPO 42g, (NH 4) 2SO 41.4g, MgSO 47H 2O 0.3g, CaCl 20.3g, CoCl 22mg, FeSO 47H 2O 5mg, MnSO 4H 2O 1.6mg, ZnSO 41.7mg, straw powder 2%, wheat bran 0.5%, water 1 000mL, 121 ℃, 0.1MPa autoclaving 30min
The cultural method of the used substratum of yeast single strain is: yeast extract paste 5g, glucose 5g, wood sugar 5g, natural pH value, water 1 000mL, 121 ℃, 0.1MPa autoclaving 30min.
The cultural method of combination fungicide preparation substratum is: straw powder and 4: 1 mixings of wheat bran mass ratio, every gram solid adds the nutritive medium of 50mL, natural pH value, 121 ℃, 0.1MPa autoclaving 30min.
The cultural method of mineral nutrition liquid is: (NH 4) 2SO 41.4g, KH 2PO 42g, urea 0.3g, MgSO 47H 2O 0.3g, CaCl 20.3g, FeSO 47H 2O 7.50mg, MnSO 4H 2O 1.50mg, ZnSO 41.0mg, CoCl 23.0mg, water 1 000mL.
The cultural method that the straw degradative rate is measured substratum is: the fresh corn stalk dries rives into pintongs, is cut into the square piece of 1cm, takes by weighing the triangular flask that 4g adds 150mL, every bottle of nutritive medium that adds 12mL again, mix, natural pH value is in 121 ℃, the following sterilization of 0.1MPa 30min.
Embodiment 4: the bacterial classification combination: with the straw degradative rate is index, and with HLG, X2, four kinds of bacterium of L3, HRQJ, concentration is 10 8Individual/mL, with 1: 1: 1: 1 volume ratio was mixed, obtained a straw degradative combination fungus strain of being made up of cellulose-degrading bacteria (HLG and X2), lignin-degrading bacteria (L3) and yeast (HRQJ), its degradation rate to stalk is significantly improved than single bacterial classification, has reached 44.75%.
At first cellulose-degrading bacteria HLG and X2 were cultivated 3 days respectively at 30 ℃, make the spore suspension that concentration is 10/mL, mix by 1: 1 volume ratio, the straw degradative rate is 38.7%, is higher than single strain; Press the same manner, HLG, X2, lignin-degrading bacteria L3 is composite, and the straw degradative rate brings up to 42.5%; Press the same manner, the HLG+X2+L3 combination is composite with yeast HRQJ, and the stalk degradation rate is increased to 44.7%.Illustrate that the bacterial strain in this combination exists certain synergetic property, does not have antagonism between them.
Embodiment 5: bacterial classification is used in combination condition and using method
1. working conditions: the field optimum inoculation amount of this straw degradative combination microbial inoculum is 4 ‰, and promptly every kilogram of stalk 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 dry straw nitrogen is 11.6 grams.Therefore the big more straw degradative that helps more of the additional amount of phosphorus need determine the phosphorus magnitude of recruitment according to practical situation.
2. using method:, in crushed stalk, spray the stalk decomposing microbial inoculum in conjunction with stalk machinery field also.Consumption: 2 kilograms every mu.Apply nitrogenous fertilizer simultaneously, every kilogram of dry straw applies 11.6 gram nitrogen, then stalk is ploughed under underground.Also applicable to the stalk fermentation of becoming thoroughly decomposed under the condition of banking up.

Claims (7)

1. the decomposition agent strain of agricultural straw fast-degradation combination is characterized in that decomposition agent strain is combined as: HLG+X2+L3+HRQJ; By HLG penicillium oxalicum (No. the 167th, the little searching of Penicillium oxalicum (2009)), X2 cellulose-degrading bacteria, L3 lignin-degrading bacteria and the unusual Pichia yeast of HRQJ (Pichia anomala; CGMCC No:2814) four kinds of fungi strains were pressed proportioning 1: 1: 1: 1 compound composition; Its preparation technology cultivates HLG, X2, four kinds of bacterium of L3, HRQJ 3 days in 30 ℃ separately, makes 10 8The spore suspension of individual/mL and bacteria suspension are respectively got 10mL and are inoculated in the fungicide preparation substratum, and inoculum size is 4%, and in 30 ℃ of cultivations, incubation time is 2-3 days, and bacterial content can reach 10 8Individual/mL.
2. the decomposition agent strain of a kind of agricultural straw fast-degradation according to claim 1 combination is characterized in that HLG, X2 in its finished product, L3, four kinds of bacterial contents of HRQJ are 10 6～10 10Individual/mL, HLG, X2, L3 bacterial strain mainly exist with spore form in the microbial inoculum.
3. the decomposition agent strain of a kind of agricultural straw fast-degradation according to claim 1 combination is characterized in that the cultural method of HLG, X2, the used substratum of L3 single strain is: K 2HPO 42g, (NH 4) 2SO 41.4g, MgSO 47H 2O 0.3g, CaCl 20.3g, CoCl 22mg, FeSO 47H 2O 5mg, MnSO 4H 2O 1.6mg, ZnSO 41.7mg, straw powder 2%, wheat bran 0.5%, water 1000mL, 121 ℃, 0.1MPa autoclaving 30min.
4. the decomposition agent strain of a kind of agricultural straw fast-degradation according to claim 1 combination is characterized in that the cultural method of the used substratum of yeast single strain is: yeast extract paste 5g, glucose 5g, wood sugar 5g, nature pH value, water 1000mL, 121 ℃, 0.1MPa autoclaving 30min.
5. the decomposition agent strain of a kind of agricultural straw fast-degradation according to claim 1 combination, it is characterized in that the cultural method that makes up the fungicide preparation substratum is: straw powder and 4: 1 mixings of wheat bran mass ratio, every gram solid adds the nutritive medium of 50mL, nature pH value, 121 ℃, 0.1MPa autoclaving 30min.
6. the decomposition agent strain of a kind of agricultural straw fast-degradation according to claim 1 combination is characterized in that the cultural method of mineral nutrition liquid is: (NH 4) 2SO 41.4g, KH 2PO 42g, urea 0.3g, MgSO 47H 2O 0.3g, CaCl 20.3g, FeSO 47H 2O 7.50mg, MnSO 4H 2O 1.50mg, ZnSO 41.0mg, CoCl 23.0mg, water 1000mL.
7. the decomposition agent strain of a kind of agricultural straw fast-degradation according to claim 1 combination, the cultural method that it is characterized in that straw degradative rate mensuration substratum is: the fresh corn stalk dries rives into pintongs, be cut into the square piece of 1cm, take by weighing the triangular flask of 4g adding 150mL, every bottle of nutritive medium that adds 12mL again, mix, natural pH value is in 121 ℃, the following sterilization of 0.1MPa 30min.
CN200910172426A 2009-09-30 2009-09-30 Agricultural straw fast-degradation decomposition agent strain composition Pending CN101838613A (en)

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CN103230917A (en) * 2013-03-14 2013-08-07 东北林业大学 Forest floor combustible biodegradation agent and use thereof
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