CN101519641A - Method for producing mixed microorganism inoculum - Google Patents
Method for producing mixed microorganism inoculum Download PDFInfo
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- CN101519641A CN101519641A CN200910029205A CN200910029205A CN101519641A CN 101519641 A CN101519641 A CN 101519641A CN 200910029205 A CN200910029205 A CN 200910029205A CN 200910029205 A CN200910029205 A CN 200910029205A CN 101519641 A CN101519641 A CN 101519641A
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Abstract
The invention relates to a method for producing a mixed microorganism solid inoculum. The method comprises the following steps: firstly, performing solid fermentation on actinomycete strains in a mixed microbial inoculum to obtain a dry solid leavening; secondly, cultivating bacterial strains through liquid fermentation respectively to obtain concentrated thalli; thirdly, fixing the asporulate bacterial concentrated thalli obtained after the fermentation by an immobilization method and preparing the same into solid particles for standby; fourthly, evenly spraying the sporulate bacterial concentrated thalli suspension obtained after the fermentation to the obtained dry solid leavening containing actinomycetes, and promoting spore bacteria to generate spores by a variable-temperature drying technique to obtain a dry inoculum; and finally, mixing the prepared dry solid particles containing the thalli to prepare the mixed microorganism inoculum. The method can produce a microorganism inoculum with various mixed functional microorganisms, effectively prevent soil-borne fungi diseases for plants, increase the output and the quality of crops, and improve soils. The production and the using method are simple, the cost is lower and the technique is easy to popularize.
Description
Technical field
The present invention relates to agricultural biological technical field, be specifically related to a kind of production method of mixing microorganisms solid inoculum.
Technical background
The relation of microorganism and plant is very close in soil, and normal and useful rhizospheric microorganism can promote plant-growth, and plays the effect that escorts in the plant-growth cycle; And plant pathogenic microorganisms can infect plant and causes disease, suppresses plant-growth even kill plants.There is the not beneficial microorganism of same-action of multiple performance in the plant rhizosphere.According to the principle that has restriction or checking relation in five elements and compatibility relation between the alternate biology, screen various functions (as the tool enzymatic activity high from plant rhizosphere, secretion growth hormone, produce microbiotic and antagonism pathogenic micro-organism and can from soil, discharge nutritive element etc.) microbial strains, utilize these functional microorganisms in the vital movement of plant rhizosphere and the effect of meta-bolites thereof, the antagonism plant pathogenic microorganisms, strengthen stress resistance of plant, fixed nutritive element in decomposition and the release soil, the secretion growth substance is realized improving output and is improved quality, reduce the use of chemical fertilizer and agricultural chemicals, improve edatope, reduce the purpose of disease sickness rate.
At present, microorganism inoculum has the Inoculant and the compound microbial inoculant of single culture.Microorganism inoculum can directly be manured into soil or be inoculated in plant rhizosphere as seed dressing in agriculture production, can also be composite in various fertilizer.Usually, be called microbial fertilizer (claiming bacterial manure again), also have products such as the biological organic fertilizer made by microorganism inoculum or compound organic and inorganic fertilizer simultaneously by microorganism and various organic fermentation or the fertilizer that is mixed and made into.Because, envrionment conditions is also had certain requirement,, could satisfy the demand of different areas Different Crop so all kinds of inoculum all should have series product according to stronger selectivity is arranged between beneficial microorganism and crop.
According to plant rhizosphere microbe ecological Studies and application and development trend thereof, microorganism inoculum is inevitable to be changed to composite bacteria by single culture.During this transfer, at first no longer merely pursue the raising of nutritive element level of supply, and to realize simultaneously multi-functional, as disease-resistant, keep away worm etc.5406 bacterial classifications are a kind of traditional zymophytes, can strengthen the resistance against diseases of crop, reduce the use of chemical pesticide, and by comparison, now the composite bacteria of using has been owing to brought into play reciprocal, collaborative, symbiosis, reinforcement or coordination effect, so the performance of bacterial classification is further improved.Secondly the microorganism inoculum of composite bacteria has prolonged the survival time of functional microorganism in soil, and the time of microbial survival is long more, and the lasting period of microbial fertilizer is long more.Microorganism length of survival time in soil depends primarily on available carbon source level in the soil, can be by in bio-fertilizer, adding the specified microorganisms bacterial strain that can decompose carbon compound in the soil, with other microbe carbon source nutrition of continuous supply, reach the effect that prolongs microorganism survival time in soil.
Microbial fertilizer has been abundant recognition and acceptance to the effect of plant, but most products effect aborning of having used at present and unstable, thereby limited it and popularized.This is because action of microorganisms is subjected to the restriction of many conditions, as: aspects such as effective quantity of the functional microorganism that the character of microbial function bacterial strain and the relation between the plant, functional microorganism are preserved in the survival time of functional microorganism on carrier and plant materials, the product in the relation of the ecological niche of plant rhizosphere, the single microbial inoculum of using or mix bacterium agent and indigenous microorganism, inoculum and application method thereof.If unclear to these restraining factors, the effect of functional microorganism just can not be given full play to, and therefore need go deep into and systematic research, establishes solid theory for the exploitation microbial fertilizer, and microbial fertilizer is widely used.
Though what bring into play leading role each period in plant-growth is superior microorganism population in the plant rhizosphere microbe group, the regulation and control edatope is the result that multiple microorganism plays a role.The investigator of home and overseas has screened a large amount of microorganism strains favourable to plant-growth from soil, these bacterial strains comprise many microbe species, different types of microorganism is not only active different, and cultural method, store method and the method for making microbial inoculum also are not quite similar, therefore, the microbial inoculum that microorganism inoculum is made can only be produced according to the characteristics of function stem, as solid fermentation microbial inoculum, solid absorption microbial inoculum and liquid fermentation agent etc.The production of these microbial inoculums mainly is at a kind of microorganism or two kinds of microorganisms.It is to be difficult to produce that the mixing microorganisms microbial inoculum of two or more microorganisms does not have certain technical measures guarantee, and particularly the production of the complete different mix bacterium agent of characteristic of several microorganisms must possess specific technology.
This laboratory is according to the effect to plant growth-promoting and diseases prevention two aspects, from plant rhizosphere soil, screened a large amount of microorganism strains, these bacterial strains have product ACC (1-amino-cyclopropane 1-carboxylic acid, the precursor of ethene) deaminase active on function, promote plant-growth and degeneration-resistant bacterial strain, have secretion tethelin bacterial strain, have secretion antagonism plant pathogenic fungi material bacterial strain, the bacterial strain of secretion antifungal protein etc. is arranged.These bacterial strains mainly concentrate in two quasi-microorganisms, i.e. actinomycetes and bacterium.The function actinomycetes strain mainly belongs to streptomycete and micromonospora two genus, and the function bacterium relates generally to genus bacillus and pseudomonas two genus.They are in the preparation of microbial inoculum or produce characteristics are respectively arranged, and gordian technique is also inequality.Streptomycete and micromonospora are made generally in fermenting agent, can form certain bacterium amount (being spore) behind the solid fermentation, dry back prolonged preservation; Genus bacillus needs liquid fermenting to produce a large amount of thalline in the thalli growth process, induce produce need behind the gemma dry could prolonged preservation; And pseudomonas is a kind of asporulate bacterium, the mass production of thalline needs liquid fermenting, a large amount of thalline prolonged preservation is the technology of a key, the cryodesiccated store method of general under study for action employing, but require the equipment complexity, the technology strictness, on producing, be difficult to use, therefore number of research projects has been made in this laboratory in this respect, explored immobilization and exsiccant method, made a large amount of pseudomonas bodies in the immobilization dry granules, reach 100% in the survival rate of preserving more than a year under the exsiccant condition.Under the production technology of research single culture microbial inoculum, for the function stem of different qualities is made the blended microbial inoculum, this lab design many covers technical scheme.Go through and constantly studied and tested in 3 years, finally formed the method for the production mixed microorganism inoculum of a cover comparative maturity, that is: solid/liquid fermentation technique and spraying/alternating temperature dry technology combines and produces the method for mixed microorganism inoculum
Summary of the invention
The purpose of this invention is to provide a kind of method of producing mixed microorganism inoculum, this microorganism inoculum is the method preparation by solid/liquid fermentation technique and spraying alternating temperature dry technology.
The method of the mixed microorganism inoculum of producing among the present invention, its feature may further comprise the steps:
(1) actinomycetes strain in the mix bacterium agent is adopted solid fermentation, get exsiccant solid fermentation thing;
(2) adopt liquid fermenting to carry out fermentation culture respectively bacterial isolates, must concentrate thalline;
(3) adopt the concentrated thalline of the asporulate bacterium that fermentation obtains in the immobilized method handle (2) to fix, make solid particulate, drying for standby;
(4) the concentrated thalline suspension of the sporiferous bacterium that fermentation in (2) is obtained obtains in (1) contains even spraying in the actinomycetic exsiccant solid material, adopts the alternating temperature dry technology to impel spore bacteria to produce gemma as early as possible again; Get dry microbial inoculum;
(5) dry solids that comprises thalline that (3) are made is sneaked in the dry microbial inoculum that makes in (4), mixes, and packing is placed siccative in the packing bag, promptly make the mixing microorganisms microbial inoculum.
The concrete operations of aforesaid method are:
(1) solid fermentation of actinomycetes strain: the slant strains of the actinomycetes function stem of the purification and rejuvenation of learning from else's experience, be inoculated into 31 ℃ of cultivation 7-10d in the rice medium, make actinomycetic spore be covered with the surface of each grain of rice, this bacterial classification is inoculated in the solid ingredient of sterilization with blending means under aseptic condition, 31 ℃ of fermentation culture 7-10d, regularly stir during the fermentation, note guaranteeing not pollution microbes, different actinomyces strains ferment respectively, and the fermentation back is poured in the aseptic square plate after mixing by 1:1, puts the loft drier drying, drying conditions is 60 ℃, can air blast when fermented product early stage is moistening, regularly stir but not air blast when the later stage is dry, fermented product is dried to till the constant weight;
(2) liquid fermenting of bacterial isolates: the slant strains of the bacterium function stem of the purification and rejuvenation of learning from else's experience, be inoculated in the triangular flask 31 ℃ of fermentation 48h and make seed liquor, seed liquor is inoculated into (prescription of substratum for optimizing) fermentation 48h in the fermentor tank, fermentation ends, centrifugal, collect and concentrate thalline.
(3) acquisition of immobilized thallus: the concentrated thalline of asporulate bacterium (Pseudomonas fluorescens etc.) of fermentation in (2) with 2% carrageenin mix, make solid particulate, drying.
(4) the bacterium liquid of above-mentioned (2) spissated spore production bacteria evenly is sprayed onto in the dry thing of making in (1) stirs, after dry 4-6 hour, oven dry is 6-12 hour in 60 ℃ of loft drier in the mid-40 ℃ of loft drier of the square plate that tiles.
(5) dry solids that comprises thalline that (3) are made is sneaked in the dry microbial inoculum that makes in (4) by a certain percentage, mixes, and packing is placed siccative in the packing bag, promptly make the mixing microorganisms microbial inoculum.
In the inventive method, the bacterium that must adopt immobilized method to make can not to produce gemma such as false pseudomonas bacillus etc. be present in the immobilization particle could be in dry microbial inoculum the preservation vigor.
Another target of the present invention provides a kind of mixed microorganism inoculum.Mixed microorganism inoculum is characterised in that, is to be made by above-mentioned (1)-(5) step.
Among the present invention, said actinomycetes strain is meant can conidiiferous all actinomycetes strains.Can from plant rhizosphere soil, screen, purifying, also can use existing bacterial strain.
Among the present invention, said bacterial isolates be meant in plant materials and rhizosphere all bacterial isolateses that screen, purifying.
Prepared microorganism inoculum is applied plant rhizosphere or seed dressing, can prevent plant silborne fungal diseases, promote plant-growth and strengthen stress resistance of plant.
By method of the present invention, can produce multiple functional microbial blended microorganism inoculum, this inoculum can effectively prevent plant silborne fungal diseases, increases crop yield and quality, improves the soil.Production and using method are simple, lower cost, and technology is easily promoted.
Description of drawings:
Fig. 1 is that solid/liquid fermentation technique and spraying/alternating temperature dry technology combine and produce the techniqueflow chart of mixed microorganism inoculum
Fig. 2 embodiment of the invention 3 effect photos: figure A and figure B are the short design sketchs of giving birth to of inoculation microbial inoculum and blank; Figure C and figure D inoculate the prevention effect figure that microbial inoculum inoculates Phytophthora capsici germ and toxin earlier; Figure E and figure F are the per plant quantity figure of inoculation microbial inoculum and blank; Figure G is the short living design sketch of inoculation microbial inoculum to Brassica campestris L seedling.
Embodiment:
Employed in the present invention term unless other explanation is arranged, generally has the implication of those of ordinary skills' common sense.
Below in conjunction with concrete preparation embodiment and Application Example, and comparable data is described the present invention in further detail.Should be understood that these embodiment just in order to demonstrate the invention, but not limit the scope of the invention by any way.
In following embodiment, various processes of Xiang Ximiaoshuing and method are not ordinary methods as known in the art.
In order to describe method of the present invention in detail, the bacterial strain that uses in following enforcement is preserved by the contriver laboratory, can provide to the public, but the bacterial strain that content of the present invention is not limited to enumerate among the embodiment.
The refined treasure of ME2:(Yan, Liu Weihong, the side is clean. and micromonospora E2 bacterial strain is to the inhibition of Phytophthora capsici and the rough determination of antimicrobial substance, plant protection journal, 2005, Vol.32 (4): 383-386.)
F53 (Wen Xiaojuan, intercalation refined treasure, Liu Weihong, Shen Ping, Wang Yong, the research of antagonism series bacillus F53 bacterial strain solid fermentation condition, Anhui agricultural sciences, 2007,35 (2): 461-463)
XG32 (Shen Ping, Liu Weihong, the refined precious * of Yan; Chen Shuanlin, XG32 bacterial strain produce the culture condition and the enzyme influence factor alive of acc deaminase, Nanjing Normal University's journal (natural science edition); 2008; Vol.31 (1): 104-108.) (Shen Ping, the refined precious * of Yan, Chen Shuanlin; Cui Xiaocan; Li Li. the active endophytic bacterium of tool acc deaminase is to the growth-promoting functions of capsicum with to the preventive and therapeutic effect of phytophthora root rot. plant protection journal, 2008, Vol.35 (1): 28-35.)
Embodiment 1:
The production of watermelon disease prevention growth-promoting microorganism inoculum
1, the preparation of functional bacterial strain:
(1) actinomycetes strain
Actinomycetes ACCC40021 buys from Chinese common micro-organisms bacterial classification administrative center; To screening from the plant rhizosphere separating payingoff bacteria, obtain strain function actinomycetes, code name is ME2.This two strains bacterium identifies that through antimicrobial spectrum withered germ of water-melon and watermelon anthrax bacteria are had higher antagonistic action, the protection effect of single strain fermentation thing artificial inoculation on seedling pathogenic bacteria reaches more than 80%, ACCC40021 has tangible growth-promoting functions to watermelon seedling stage, and ACCC40021 and ME2 all can be colonizated in the watermelon rhizosphere.
(2) bacterial isolates:
To in roots of plants, screening with the rhizosphere isolated bacterial, obtain two strain function bacteriums, code name is respectively F53 and XG32.Wherein XG32 is a Pseudomonas fluorescens, separates in pepper root; F53 is a series bacillus, separates from the eggplant rhizosphere.The XG32 bacterium has the acc deaminase activity, watermelon is had short giving birth to and degeneration-resistant effect, and can be colonizated in the watermelon root.F53 has antagonism various plants pathogenic bacteria and promotes the effect of plant-growth, can be colonizated in the watermelon rhizosphere.
2, the preparation of culture of strains and single microbial inoculum
(1) after sandy soil is guaranteed the actication of culture of the actinomycetes ACCC40021 that deposits and ME2, be inoculated into 31 ℃ of cultivation 7-10d in the rice medium respectively, make actinomycetic spore be covered with the surface of each grain of rice, this bacterial classification is inoculated in the solid ingredient of sterilization with blending means under aseptic condition, 31 ℃ of fermentation culture 7-10d, regularly stir during the fermentation, note guaranteeing not pollution microbes.ACCC40021 and ME2 bacterial strain ferment respectively, and the fermentation back is poured in the aseptic square plate after mixing by 1:1, puts the loft drier drying, drying conditions is 60 ℃, can air blast when fermented product early stage is moistening, regularly stir but not air blast when the later stage is dry, fermented product is dried to till the constant weight; Detect each bacterial strain quantity.
(2) learn from else's experience purification and rejuvenation bacterium F53 and the slant strains of XG32 function stem, be inoculated in the triangular flask 31 ℃ of fermentation 48h respectively and make seed liquor, seed liquor is inoculated into (prescription of substratum for optimizing) fermentation 48h in the fermentor tank, fermentation ends, centrifugal, collect and concentrate thalline.
(3) (2) fermentation XG32 bacterial strain (Pseudomonas fluorescens) concentrated thalline with 2% carrageenin mix, make solid particulate, in talcum powder, be dried to weight.That detects dry microbial inoculum contains bacterium quantity.
(4) the spissated bacterium liquid of F53 (series bacillus) is sprayed onto in the dry actinomycetes microbial inoculum of making in (1) uniformly stirs, in the mid-40 ℃ of loft drier of the square plate that tiles after dry 4-6 hour, make DP1 a large amount of gemma that form in 4-6 hour, when the thalline of the F53 bacterial strain 80% in the microbial inoculum had formed gemma after testing, 60 ℃ of loft drier were dried 6-12 hour.
3, the production of mix bacterium agent and packing
Dry microbial inoculum and the immobilized microbial inoculum of XG32 bacterial strain that actinomycetes ACCC40021 and ME2, bacterium F53 bacterial strain are made mix by a certain percentage, and the quantity of every strain bacterium is 10
6-10
8Cfu/g.Pack after indicating the thalline quantity of every kind of microorganism in the dry microbial inoculum of blended, place siccative in the packing bag, promptly make the mixing microorganisms microbial inoculum.
4, effect:
It is obvious that the water extract of microbial inoculum suppresses the watermelon blight effect, and microbial inoculum adds 5 times water droplet and adds that antibacterial circle diameter reaches 1.2cm in the cuvette of Oxford.Watermelon artificial inoculation on seedling watermelon blight preventive effect is more than 90%.The short fruit of coming into force is obvious.Effect of inoculation sees Table 1.
Table 1 mixing microorganisms Inoculant is to the prevention effect of watermelon artificial inoculation on seedling watermelon blight
Embodiment 2:
The production of strawberry disease prevention growth-promoting microorganism inoculum, flow process as shown in Figure 1.
1, the preparation of functional bacterial strain:
(1) actinomycetes strain
Actinomycetes ACCC40021 and ME2.This two strains bacterium identifies that through antimicrobial spectrum Strawberry Fusarium Wilt and strawberry anthrax bacteria are had higher antagonistic action, the protection effect of single strain fermentation thing artificial inoculation on seedling pathogenic bacteria reaches more than 80%, ACCC40021 has tangible growth-promoting functions to strawberry in seedling stage, and ACCC40021 and ME2 all can be colonizated in the strawberry rhizosphere.
(2) bacterial isolates:
From capsicum rhizosphere separation screening to a strain bacterium XG32, through being accredited as Pseudomonas fluorescens; The F53 that screens from the eggplant rhizosphere is a series bacillus, and the XG32 bacterium has higher acc deaminase activity, strawberry is had short giving birth to and degeneration-resistant effect, and can be colonizated in the strawberry root.F53 has antagonism various plants pathogenic bacteria and promotes the effect of plant-growth, can be colonizated in the strawberry rhizosphere.
2, the preparation of culture of strains and single microbial inoculum
(1) after sandy soil is guaranteed the actication of culture of the actinomycetes ACCC40021 that deposits and ME2, be inoculated into 31 ℃ of cultivation 7-10d in the rice medium respectively, make actinomycetic spore be covered with the surface of each grain of rice, this bacterial classification is inoculated in the solid ingredient of sterilization with blending means under aseptic condition, 31 ℃ of fermentation culture 7-10d, regularly stir during the fermentation, note guaranteeing not pollution microbes.ACCC40021 and ME2 bacterial strain ferment respectively, and the fermentation back is poured in the aseptic square plate after mixing by 1:1, puts the loft drier drying, drying conditions is 60 ℃, can air blast when fermented product early stage is moistening, regularly stir but not air blast when the later stage is dry, fermented product is dried to till the constant weight; Detect each bacterial strain quantity.
(2) learn from else's experience purification and rejuvenation bacterium F53 and the slant strains of XG32 function stem, be inoculated in the triangular flask 31 ℃ of fermentation 48h respectively and make seed liquor, seed liquor is inoculated into (prescription of substratum for optimizing) fermentation 48h in the fermentor tank, fermentation ends, centrifugal, collect and concentrate thalline.
(3) (2) fermentation the XG32 bacterial strain concentrated thalline with 2% carrageenin mix, make solid particulate, in talcum powder, be dried to weight.That detects dry microbial inoculum contains bacterium quantity.
(4) the spissated bacterium liquid of F53 (series bacillus) is sprayed onto in the dry actinomycetes microbial inoculum of making in (1) uniformly stirs, in the mid-40 ℃ of loft drier of the square plate that tiles after dry 4-6 hour, make DP1 a large amount of gemma that form in 4-6 hour, when the thalline of the F53 bacterial strain 80% in the microbial inoculum had formed gemma after testing, 60 ℃ of loft drier were dried 6-12 hour.
3, the production of mix bacterium agent and packing
Dry microbial inoculum and the immobilized dry microbial inoculum of XG32 bacterial strain that actinomycetes ACCC40021 and ME2, F53 bacterial strain are made mix by a certain percentage, pack after indicating the thalline quantity of every kind of microorganism in the dry microbial inoculum of blended, place siccative in the packing bag, promptly make the mixing microorganisms microbial inoculum.
4, effect:
It is obvious that the water extract of microbial inoculum suppresses strawberry blight effect, and microbial inoculum adds 5 times water droplet and adds that antibacterial circle diameter reaches 1.2cm in the cuvette of Oxford.Strawberry artificial inoculation on seedling watermelon blight preventive effect is more than 70%.The short fruit of coming into force obviously sees Table 2
Table 2 mixing microorganisms Inoculant is to the influence of strawberry seedling growth
Embodiment 3:
The production of vegetables disease prevention growth-promoting microorganism inoculum
1, the preparation of functional bacterial strain:
(1) actinomycetes strain
Actinomycetes ACCC40021 and ME2.This two strains bacterium identifies that through antimicrobial spectrum wilt and anthrax bacteria to various vegetables have higher antagonistic action, the protection effect of single strain fermentation thing artificial inoculation on seedling pathogenic bacteria reaches more than 80%, ACCC40021 has tangible growth-promoting functions to capsicum in seedling stage, and ACCC40021 and ME2 all can be colonizated in plant rhizospheres such as capsicum and tomato.
(2) bacterial isolates:
From eggplant rhizosphere separation screening to a strain bacterium F53, through being accredited as series bacillus; This strain bacterium has short giving birth to and resistant effect to various vegetables, and can be colonizated in the various vegetables rhizosphere.
2, the preparation of culture of strains and single microbial inoculum
(1) after sandy soil is guaranteed the actication of culture of the actinomycetes ACCC40021 that deposits and ME2, be inoculated into 31 ℃ of cultivation 7-10d in the rice medium respectively, make actinomycetic spore be covered with the surface of each grain of rice, this bacterial classification is inoculated in the solid ingredient of sterilization with blending means under aseptic condition, 31 ℃ of fermentation culture 7-10d, regularly stir during the fermentation, note guaranteeing not pollution microbes.ACCC40021 and ME2 bacterial strain ferment respectively, and the fermentation back is poured in the aseptic square plate after mixing by 1:1, puts the loft drier drying, drying conditions is 60 ℃, can air blast when fermented product early stage is moistening, regularly stir but not air blast when the later stage is dry, fermented product is dried to till the constant weight; Detect each bacterial strain quantity.
(2) the learn from else's experience slant strains of bacterium F53 function stem of purification and rejuvenation is inoculated into that 31 ℃ of fermentation 48h make seed liquor in the triangular flask, and seed liquor is inoculated into (prescription of substratum for optimizing) fermentation 48h in the fermentor tank, and fermentation ends is centrifugal, collects and concentrates thalline.
(3) the spissated bacterium liquid of F53 is sprayed onto in the dry actinomycetes microbial inoculum of making in (1) uniformly stirs, in the mid-40 ℃ of loft drier of the square plate that tiles after dry 4-6 hour, make F53 a large amount of gemma that form in 4-6 hour, when the thalline of the F53 bacterial strain 80% in the microbial inoculum had formed gemma after testing, 60 ℃ of loft drier were dried 6-12 hour.
3, the production of mix bacterium agent and packing
The dry mixed microbial inoculum that actinomycetes ACCC40021, ME2, F53 bacterial strain make is indicated the wherein thalline quantity of every kind of microorganism, and packing is placed siccative in the packing bag then, promptly makes the mixing microorganisms microbial inoculum.
4, effect:
The water extract of microbial inoculum suppress cucumber fusarium axysporum and eggplant verticillium wilt bacterium effect obvious, microbial inoculum adds 5 times water droplet and adds that antibacterial circle diameter reaches 1.2cm in the cuvette of Oxford.Eggplant artificial inoculation on seedling verticillium wilt pathogen preventive effect is more than 70%, to the capsicum epidemic disease preventive effect more than 80%.The inoculation preventive effect is seen diagram.Composite fungus agent comes into force fruit obviously to capsicum is short, and effect of increasing production is obvious.See Fig. 2 and table 3.
Table 3 mixing microorganisms Inoculant is to the influence of yield of hot pepper
Claims (4)
1, a kind of production method of mixed microorganism inoculum is characterized in that,
(1) actinomycetes strain in the mix bacterium agent is adopted solid fermentation, get exsiccant solid fermentation thing;
(2) adopt liquid fermenting to carry out fermentation culture respectively bacterial isolates, must concentrate thalline;
(3) adopt the concentrated thalline of the asporulate bacterium that fermentation obtains in the immobilized method handle (2) to fix, make solid particulate, drying for standby;
(4) the concentrated thalline suspension of the sporiferous bacterium that fermentation in (2) is obtained obtains in (1) contains even spraying in the actinomycetic exsiccant solid material, adopts the alternating temperature dry technology to impel spore bacteria to produce gemma as early as possible again; Get dry microbial inoculum;
(5) dry solids that comprises thalline that (3) are made is sneaked in the dry microbial inoculum that makes in (4), mixes, and packing is placed siccative in the packing bag, promptly make the mixing microorganisms microbial inoculum.
2, the production method of mixed microorganism inoculum according to claim 1 is characterized in that, concrete operations are:
(1) solid fermentation of actinomycetes strain: the slant strains of the purification and rejuvenation of learning from else's experience, be inoculated into and cultivate 7-10d in the rice medium, make actinomycetic spore be covered with the surface of each grain of rice, this bacterial classification is inoculated in the solid ingredient of committing genocide with blending means under aseptic condition, 31 ℃ of fermentation culture 7-10d, regularly stir during the fermentation, note guaranteeing not pollution microbes, different actinomyces strains ferment respectively, and the fermentation back is poured in the aseptic square plate after mixing by 1:1, puts the loft drier drying, drying conditions is 60 ℃, can air blast when fermented product early stage is moistening, regularly stir but not air blast when the later stage is dry, fermented product is dried to till the constant weight;
(2) liquid fermenting of bacterial isolates: the slant strains of the purification and rejuvenation of learning from else's experience, be inoculated in the triangular flask 31 ℃ of fermentation 48h respectively and make seed liquor, seed liquor is inoculated into fermentation cylinder for fermentation 48h, and fermentation ends is centrifugal, collects to concentrate thalline;
(3) acquisition of immobilized thallus: the concentrated thalline of asporulate bacterium of fermentation in (2) with 2% carrageenin mix, make solid particulate, drying;
(4) above-mentioned (2) spissated bacterium liquid is sprayed onto in the dry thing of making in (1) uniformly stirs, after dry 4-6 hour, the temperature of loft drier is transferred to 60 ℃ of oven dry 6-12 hour in the mid-40 ℃ of loft drier of the square plate that tiles;
(5) dry solids that comprises thalline that (3) are made is sneaked in the dry microbial inoculum that makes in (4), mixes, and packing is placed siccative in the packing bag, promptly make the mixing microorganisms microbial inoculum.
3, a kind of is the mixed microorganism inoculum that is made by claim 1 or 2 described methods.
4, mixed microorganism inoculum according to claim 3 is characterized in that, the mixing microorganisms microbial inoculum is a pressed powder, includes actinomycetes and bacterium, and the quantity of every strain bacterium is 10
6-10
8Cfu/g.
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| CN111411046A (en) * | 2020-04-17 | 2020-07-14 | 中国矿业大学(北京) | Dark color endophytic fungus agent and application thereof |
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| CN1511940A (en) * | 2002-12-31 | 2004-07-14 | 福建农林大学 | Compound microbe fermentation strain preparing method and its use |
| CN1216014C (en) * | 2003-08-21 | 2005-08-24 | 中国农业科学院油料作物研究所 | Fixed bacterial fertilizer and preparing method thereof |
-
2009
- 2009-04-03 CN CN2009100292051A patent/CN101519641B/en not_active Expired - Fee Related
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| CN102010825A (en) * | 2011-01-20 | 2011-04-13 | 上海创博生态工程有限公司 | Microorganism compound bactericide for controlling cucumber fusarium wilt and preparation method thereof |
| CN102010825B (en) * | 2011-01-20 | 2013-05-29 | 上海创博生态工程有限公司 | Microorganism compound bactericide for controlling cucumber fusarium wilt and preparation method thereof |
| CN105916986A (en) * | 2013-10-08 | 2016-08-31 | 日本农药株式会社 | Inoculum formed by inoculating microorganism, and method for producing antibiotic using same |
| CN108179130A (en) * | 2018-03-21 | 2018-06-19 | 广州同心源生物科技有限公司 | A kind of preparation method of high activity Enterococcus faecalis microorganisms preparation dry powder |
| CN108432425A (en) * | 2018-04-17 | 2018-08-24 | 黄昱子 | Filter paper bag anther fertilizer |
| CN111411046A (en) * | 2020-04-17 | 2020-07-14 | 中国矿业大学(北京) | Dark color endophytic fungus agent and application thereof |
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| CN101519641B (en) | 2011-07-20 |
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