CN106244491A - A kind of banana stem leaves residuum decomposing microbial inoculum suppressing droop and preparation method thereof - Google Patents
A kind of banana stem leaves residuum decomposing microbial inoculum suppressing droop and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of banana stem leaves residuum decomposing microbial inoculum suppressing droop and preparation method thereof, it is characterized in that with having cellulose, lignin, the dirty flat lead fungi of chromogen hair of hemicellulose decomposition function, Fructus Citri tangerinae green trichoderma, aspergillus oryzae, and there is Biological control function bacillus amyloliquefaciens, general streptomycete composite fermentation of receiving is made.This decomposing microbial inoculum can accelerate the rotten speed of also field banana stem leaves residuum effectively, can suppress the growth of the pathogen such as banana blight simultaneously.After this decomposing microbial inoculum is used in field, banana stem leaves Residue decay speed is accelerated, and wherein banana stem leaves residuum weight-loss ratio dramatically increases, banana stem leaves residuum pulling force significantly reduces;Soil surface characters antagonistic Bacillus dramatically increases with Population of Actinomycetes, and pathogenic microbes quantity significantly reduces.
Description
Technical field
Patent of the present invention relates to a kind of banana stem leaves residuum decomposing microbial inoculum suppressing droop and preparation method thereof, belongs to agriculture
Industry technical field of microbe application.
Background technology
Owing to Fructus Musae has fast-growing and Biomass high, while producing Fructus Musae, also produce the Fructus Musae being close to equivalent
Haulm by-product, it is estimated that often produce 1hm2Fructus Musae can produce 600-900 ton stem stalk, and its resource is the abundantest.And banana stem leaves
As a kind of special agricultural wastes, compared with other agricultural crop straw, containing abundant nutritional labeling, in banana stem leaves
Carbohydrate content is high, and according to another report, every strain Fructus Musae remains in the nitrogen average out to 57 in cauloid, blade, bulb and root system
Gram, phosphorus element is 6.6 grams, potassium element 232 grams, if average utilization is respectively based on 22%, 4% and 46.22%, and the utilization in stem and leaf also field
Rate is based on 30%, then the ammonia in per hectare banana stem leaves also field, phosphorus, potassium content are respectively equivalent to 1.29 tons, 0.8 ton, 0.75 ton and answer
Hefei (content 3 15%).Major part banana plantation, Guangdong Province at present, banana stem leaves is often dropped the edge of a field or river, not only
Cause the waste of nutrients resource in stem and leaf, and the pathogenic bacteria caught an illness in Fructus Musae residuum is flowed by water system and more easily causes pathogenic bacteria
Propagate.As can be seen here, carry out Fructus Musae and discard the recycling research of stem and leaf resource, have wide development and application space, to perfume (or spice)
The sustainable development of any of several broadleaf plants industry also has important realistic meaning.
Containing abundant nutritional labeling in banana stem leaves, and it is extremely low that Fructus Musae discards utilization rate in stem at present, the most former
Because being that its volume is big, moving difficulty, content of cellulose is high, it is difficult to decompose at short notice, therefore mostly as garbage quilt
Abandon, not only cause the great wasting of resources, and pollute environment, bring a lot of trouble also to any of several broadleaf plants field management subsequently.
In order to not make banana stem leaves waste, China's researcher has also made more research, and Kuang Shi is grown and waited in patent
Disclose in numbers 2010102334329 and a kind of utilize banana stem leaves to carry out the method that biofermentation makes biological organic fertilizer: will cut
Any of several broadleaf plants bar fallen axially cuts away half, and allows its natural drying 5-7 days, with pulverizer or hay cutter, any of several broadleaf plants bar is ground into a length of 2-3
Centimetre pipe nipple, add chicken manure and biofermentation microbial inoculum, mixing stirs evenly, and piles up holding 25-30 days, carries out fermentation and form.This side
Method improves the utilization rate of banana stem leaves really, but its production work amount is that comparison is huge, and the transport of banana stem leaves also will be relatively
Many is artificial；Chen Renjie etc. disclose one in patent No. 201310609319X and utilize banana stem leaves making beef cattle biology to raise
The method of material: after banana stem leaves is pulverized by it, after allocating adjuvant into, also carried out sterilization treatment, energy resource consumption with high steam
Bigger
Han Lina etc. disclose a kind of banana stem organic fertilizer and preparation method thereof in patent No. 201110272946X: main
It is as the organic method carrying out fermenting with muck using banana stem.He Han etc. are disclosed in the patent No. 2012100551946
A kind of method utilizing banana stalks, stem and leaf to make biological organic fertilizer: with banana stalks, stem and leaf be main work raw material, with animal wastes,
Pond sludge, for adjuvant, forms with the EM strain fermentation of multistage expansion.These fermentation process are the fermentation process of traditional fertilizer,
Its fermentation high temperature can suppress or kill pathogen.
But the existing method utilizing banana stem leaves is substantially that dystopy (not carrying out at former planting site) processes, and does not also have
Have and fundamentally solve its problem moved, and in transportation, also result in the pathogen (such as droop) of some plant
Diffusion and cause the generation of other planting site diseases.Therefore, if to become thoroughly decomposed the most on the spot and just must solve two
Individual problem: one is quickly by the degraded of the fiber in banana stem leaves；Two is suppressing or killing of the pathogen in banana stem leaves
Problem.
For problem above, the present invention uses microbial strains and the efficiently suppression with efficient degradation banana stem leaves ability
The biocontrol strains of banana blight makes decomposing agent, and this decomposing agent can be bred a large amount of during decomposing banana stem leaves
Biocontrol strains, the effective sprouting suppressing soil-borne pathogen.
Summary of the invention
For the problems referred to above, the invention discloses a kind of banana stem leaves residuum decomposing microbial inoculum suppressing droop, originally become thoroughly decomposed
Microbial inoculum by having stronger cellulose, lignin, the dirty flat lead fungi of chromogen hair of hemicellulose decomposition function, Fructus Citri tangerinae green trichoderma, aspergillus oryzae,
With there is Biological control function bacillus amyloliquefaciens, general streptomycete composite fermentation of receiving is made.This decomposing microbial inoculum can be effectively
Accelerate the rotten speed of also field banana stem leaves residuum, the growth of the pathogen such as banana blight can be suppressed simultaneously.Field is used
After this decomposing microbial inoculum banana stem leaves Residue decay speed accelerate, wherein banana stem leaves residuum weight-loss ratio dramatically increase, banana stem leaves
Residuum pulling force significantly reduces；Soil surface characters antagonistic Bacillus dramatically increases with Population of Actinomycetes, pathogenic microbes quantity
Significantly reduce.
In order to achieve the above object, the invention discloses the system of a kind of banana stem leaves residuum decomposing microbial inoculum suppressing droop
Preparation Method, its concrete preparation method, comprise the following steps:
Step one, the preparation of compound aspergillosis mycopowder
A, the preparation of dirty chromogen hair flat lead fungi seed liquor: take out dirty chromogen hair flat lead fungi culture presevation pipe, use PDA solid medium
Draw flat board to recover, cultivate 7 days for 30 DEG C.Under flat board, the streak inoculation of picking single bacterium colony is equipped with 150 milliliters of PDA solid culture
In the Fructus Solani melongenae bottle of base, cultivate 6-8 days for 30 DEG C in incubator, treat that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore powder and i.e. can use 500
The physiological saline solution eluting of milliliter, regulation spore concentration is 0.01 hundred million cfu/ml, is dirty chromogen hair flat lead fungi seed liquor；
B, the preparation of Fructus Citri tangerinae green trichoderma seed liquor: take out Fructus Citri tangerinae green trichoderma bacterium culture presevation pipe, draw flat board with PDA solid medium and carry out
Recovery, cultivates 6 days for 30 DEG C.Under flat board, the streak inoculation of picking single bacterium colony is equipped with the Fructus Solani melongenae bottle of 150 milliliters of PDA solid mediums
In, cultivate 4-6 days for 30 DEG C in incubator, treat that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore and i.e. can use the aseptic life of 500 milliliters
Reason salt water, regulation spore concentration is 0.1 hundred million cfu/ml, is Fructus Citri tangerinae green trichoderma seed liquor；
C, the preparation of aspergillus oryzae seed liquor: take out aspergillus oryzae strain preservation pipe, draw flat board with PDA solid medium and recover,
Cultivate 5 days for 30 DEG C.Under flat board, the streak inoculation of picking single bacterium colony is equipped with in the Fructus Solani melongenae bottle of 150 milliliters of PDA solid mediums,
Incubator is cultivated 5-7 days for 30 DEG C, treats that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore and i.e. can use the sterile physiological salt of 500 milliliters
Water elution, regulation spore concentration is 100,000,000 cfu/ml, is aspergillus oryzae seed liquor；
D, mixed fermentation: by the dirty chromogen hair flat lead fungi seed liquor of above-mentioned preparation, Fructus Citri tangerinae green trichoderma seed liquor and aspergillus oryzae seed liquor
After the ratio mix homogeneously of 2:1:1, being seeded in the aspergillosis solid fermentation culture medium of sterilizing with the inoculum concentration of 2%, strain is with solid
After body culture medium mix homogeneously, being spread out in connecting the aspergillosis solid fermentation culture medium planted on fermentation tank, height of materials is 5-10cm,
Temperature control 28-32 DEG C, humidity remains 60%-75%, ferments 6-10 days, detects its total spore content and is not less than 2,000,000,000 CFU/g, detection
Its dirty chromogen hair flat lead fungi spore content is not less than 1,000,000,000 CFU/g, and lignin peroxidase is lived, i.e. LiP enzyme is lived and is not less than
200U/ g；Manganese-dependent peroxidase, i.e. MnP enzyme is lived and is not less than 500U/ g；Cellulase content is not less than 500IU/ g, wood
Dextranase content the lowest 1000IU/ g, gets final product low-temperature air-drying, and moisture is less than 10%, i.e. obtains compound aspergillosis mycopowder；
Wherein, described PDA solid medium: Rhizoma Solani tuber osi 200g, sucrose 20g, water 1000mL, agar 20g；
Wherein, described aspergillosis solid fermentation culture medium: banana stem leaves slag 85%, cottonseed meal 2%, corn cob 8%, stone powder 1%, thick Semen Maydis
Skin 2%, carbamide 0.4%, dipotassium hydrogen phosphate 0.4%, magnesium sulfate 0.05%, moisture content in medium 50%-60%.The bar of each medium sterilization
Part is: 0.10-0.15MPa, 121 DEG C of sterilizings 30 minutes.
Step 2, the preparation of bacillus amyloliquefaciens mycopowder
Take out bacillus amyloliquefaciens preservation pipe, draw flat board respectively with nutrient solid medium and recover, 30 DEG C of cultivations
48 hours.Under flat board, the single colony inoculation of picking is to equipped with nutrient solid medium, cultivates 48 in 30 DEG C of incubators
Hour, with 3000ml sterilized water by the lawn eluting in three Fructus Solani melongenae bottles, it is seeded to equipped with 300L liquid seed culture medium
In 500L fermentation tank, opening stirring 120r/min, within first 10 hours, ventilation is 200L/min, and after 10 hours, ventilation is 320L/min
, cultivate 16-24 hour, treat that total bacterial content is not less than 2,000,000,000 cfu/ml, can be used as bacillus cereus liquid seed for 30 DEG C；
Fermentation: the bacillus cereus liquid seed of above-mentioned preparation is added in spore solid fermentation culture medium, mixing, tray is sent out
Ferment, windrow thickness is 5-10cm, controls fermentation temperature and is 30-40 DEG C, ferments 36-48 hour, treat that spore content is not less than 10,000,000,000
Cfu/g, can dry, and treats that moisture is less than 10%, i.e. obtains bacillus amyloliquefaciens mycopowder；
Wherein, described nutrient solid medium: peptone 5g, Carnis Bovis seu Bubali cream 3g, sodium chloride 5g, agar 20 g, water 1000mL,
pH7.2；
Wherein, described liquid seed culture medium: glucose 8g/L, Semen Maydis powder 10 g/L, Semen Glycines powder 10 g/L, magnesium sulfate 0.2 g/
L, manganese sulfate 0.1 g/L, potassium dihydrogen phosphate 0.5 g/L, disodium hydrogen phosphate 1.0 g/L, pH7.0.The condition of each medium sterilization
For 0.10-0.15MPa, 121 DEG C of sterilizings 30 minutes；
Wherein, spore solid fermentation culture medium: banana stem leaves slag 60%, wheat bran 35%, cottonseed meal 2%, stone powder 1%, thick skin of Semen Maydis 2%, urine
Element 0.4%, dipotassium hydrogen phosphate 0.4%, magnesium sulfate 0.05%, moisture content in medium 50%-60%.The condition of each medium sterilization is:
0.10-0.15MPa, 121 DEG C of sterilizings 30 minutes.
Step 3, the general preparation receiving streptomycete mycopowder
Take out general streptomyces species preservation pipe of receiving, draw flat board with Gause I solid medium and recover, cultivate 7 days for 30 DEG C.
Under flat board, the streak inoculation of picking single bacterium colony is equipped with in the Fructus Solani melongenae bottle of 150 milliliters of Gause I solid mediums, at incubator
In 30 DEG C cultivate 6-8 days, treat that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore powder and i.e. can use the physiological saline solution of 500 milliliters to wash
De-, regulation spore concentration is 0.1 hundred million cfu/ml, is general streptomycete seed liquor of receiving；
Fermentation: the general streptomycete solid of receiving that streptomycete seed liquor of above-mentioned preparation general being received is seeded to sterilizing with the inoculum concentration of 5% is sent out
In ferment culture medium, strain is received after streptomycete solid medium mixs homogeneously with general, the general streptomycete solid fermentation of receiving that will connect kind
Culture medium is spread out on fermentation tank, and height of materials is 5-8cm, temperature control 28-32 DEG C, and a few days ago air humidity remains 60%-75%, mistake
Rear air humidity remains 40%-50%, ferments 4-6 days, detects its spore content and is not less than 1,000,000,000 CFU/g, gets final product low-temperature air-drying,
I.e. obtain general streptomycete mycopowder of receiving；
Wherein, described Gause I solid medium: potassium nitrate: 1 gram, soluble starch: 20 grams, dipotassium hydrogen phosphate: 05
Gram, magnesium sulfate: 05 grams, sodium chloride: 05 grams, ferrous sulfate: 0 01 grams, agar: 20 grams, distilled water supplies 1000ml, PH7
2～7 4；
Wherein, described general streptomycete solid fermentation culture medium of receiving: banana stem leaves slag 80%, cottonseed meal 2%, Semen Maydis powder 1%, stone powder
10%, wheat bran 7%, manganese sulfate 0.004%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate 0.05%, solid medium water content is 40%-60%.
The condition of each medium sterilization is: 0.10-0.15MPa, 121 DEG C of sterilizings 30 minutes.
Step 4 by compound aspergillosis mycopowder, bacillus amyloliquefaciens mycopowder, general receive streptomycete mycopowder with cottonseed meal powder with 1:1:
The ratio mix homogeneously of 1:1, detects its dirty chromogen hair flat lead fungi spore content and is not less than 200,000,000 CFU/g, lignin peroxidase
Live and be not less than 40U/ g；Manganese-dependent peroxidase is lived and is not less than 100U/ g；Cellulase content is not less than 100IU/ g, wood
Dextranase content the lowest 200IU/ g, solves starch spore content and is not less than 2,000,000,000 CFU/g, and general streptomycete content of receiving is not less than 200,000,000
CFU/g, gets final product packed products.
Wherein, the using method of the banana stem leaves residuum decomposing microbial inoculum of described suppression droop is by 1-5kg decomposing microbial inoculum
Dilute 100 times uniformly to spray to 1 mu of banana stem leaves residuum；Alternatively, it is also possible to by 2-10kg decomposing microbial inoculum/every mu, entirely
Ground is uniformly spread in banana stem leaves residuum.
Beneficial effects of the present invention:
Containing white rot fungi in the present invention--the dirty flat lead fungi of chromogen hair, it can produce abundant lignin-degrading enzymes (lignin mistake
Oxide enzyme；Manganese-dependent peroxidase), thus fast degradation banana stem leaves residuum.Decomposing microbial inoculum in the present invention is the most former
Material utilizes the fermentation of banana stem leaves residuum to form, the suitably Fast-propagation life in banana stem leaves residuum of all of bacterial strain
Long, decrease each strain and do not grow due to cause in different substrates or breed slow risk；Strain in the present invention is
By screening test for many years, after various strains apply in banana stem leaves residuum, first by the dirty flat lead fungi of chromogen hair, the green wood of Fructus Citri tangerinae
Mould, that aspergillus oryzae produces all kinds of cellulase, lignoenzyme, the cellulose in glucanase degraded banana stem leaves residuum, wooden
The carbohydrates such as element become low molecule saccharide, for bacillus amyloliquefaciens, general receive streptomycete and utilize growth, and produce a large amount of
Antagonistic substance, thus suppress or kill Fusarium oxysporum in soil, and promote the growth and breeding of probiotics.
5 strain bacterium in this decomposing microbial inoculum can in banana stem leaves residuum symbiotic co-existence, it is possible to effectively accelerate also field
The rotten speed of banana stem leaves residuum, can suppress the growth of the pathogen such as banana blight simultaneously.This bacterium of becoming thoroughly decomposed is used in field
After agent banana stem leaves Residue decay speed accelerate, wherein banana stem leaves residuum weight-loss ratio dramatically increase, banana stem leaves residuum pulling force
Significantly reduce;Antagonism bud pole bacterium dramatically increases soil surface characters with Population of Actinomycetes, and pathogenic microbes quantity significantly reduces.
Products and marketing product of the present invention compares and has the advantage that using Fusarium oxysporum quantity in rear soil only executes
The 10%-20% processed with conventional commercial decomposing agent.After using this decomposing agent, soil suppresses the bacillus cereus number of Fusarium oxysporum
Amount is relatively used conventional decomposing agent and is processed increase 1000-10000 times；The Population of Actinomycetes suppressing Fusarium oxysporum in soil is relatively used
Conventional decomposing agent processes and adds 100-1000 times.
Detailed description of the invention
Embodiment 1
A kind of banana stem leaves residuum decomposing microbial inoculum suppressing droop, it is characterised in that the banana stem leaves of described suppression droop
Residuum decomposing agent is mainly by following several bacterium: the dirty flat lead fungi of chromogen hair, Fructus Citri tangerinae green trichoderma, aspergillus oryzae, bacillus amyloliquefaciens, Pu Na
Streptomycete fermentation is made；Its concrete preparation method, comprises the following steps:
Step one, the preparation of compound aspergillosis mycopowder
A, the preparation of dirty chromogen hair flat lead fungi seed liquor: take out dirty chromogen hair flat lead fungi culture presevation pipe, use PDA solid medium
Drawing flat board to recover, cultivate 7 days for 30 DEG C, under flat board, the streak inoculation of picking single bacterium colony is equipped with 150 milliliters of PDA solid culture
In the Fructus Solani melongenae bottle of base, cultivate 6-8 days for 30 DEG C in incubator, treat that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore powder and i.e. can use 500
The physiological saline solution eluting of milliliter, regulation spore concentration is 0.01 hundred million cfu/ml, is dirty chromogen hair flat lead fungi seed liquor；
B, the preparation of Fructus Citri tangerinae green trichoderma seed liquor: take out Fructus Citri tangerinae green trichoderma bacterium culture presevation pipe, draw flat board with PDA solid medium and carry out
Recovery, cultivates 6 days for 30 DEG C, and under flat board, the streak inoculation of picking single bacterium colony is equipped with the Fructus Solani melongenae bottle of 150 milliliters of PDA solid mediums
In, cultivate 4-6 days for 30 DEG C in incubator, treat that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore and i.e. can use the aseptic life of 500 milliliters
Reason salt water, regulation spore concentration is 0.1 hundred million cfu/ml, is Fructus Citri tangerinae green trichoderma seed liquor；
C, the preparation of aspergillus oryzae seed liquor: take out aspergillus oryzae strain preservation pipe, draw flat board with PDA solid medium and recover,
Cultivating 5 days for 30 DEG C, under flat board, the streak inoculation of picking single bacterium colony is equipped with in the Fructus Solani melongenae bottle of 150 milliliters of PDA solid mediums,
Incubator is cultivated 5-7 days for 30 DEG C, treats that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore and i.e. can use the sterile physiological salt of 500 milliliters
Water elution, regulation spore concentration is 100,000,000 cfu/ml, is aspergillus oryzae seed liquor；
D, mixed fermentation: by the dirty chromogen hair flat lead fungi seed liquor of above-mentioned preparation, Fructus Citri tangerinae green trichoderma seed liquor and aspergillus oryzae seed liquor
After the ratio mix homogeneously of 2:1:1, being seeded in the aspergillosis solid fermentation culture medium of sterilizing with the inoculum concentration of 2%, strain is with solid
After body culture medium mix homogeneously, being spread out in connecting the aspergillosis solid fermentation culture medium planted on fermentation tank, height of materials is 5-10cm,
Temperature control 28-32 DEG C, humidity remains 60%-75%, ferments 8 days, and detecting its total spore content is 3,000,000,000 CFU/g, detects its dirty color
Raw wool flat lead fungi spore content is 1,300,000,000 CFU/g, and lignin peroxidase is lived, i.e. LiP enzyme is lived as 265U/ g；Manganese relied on
Oxide enzyme, i.e. MnP enzyme is lived as 621U/ g；Cellulase content is 788IU/ g, and xylanase content is 1508IU/ g, i.e.
Can low-temperature air-drying, moisture is less than 10%, i.e. obtains compound aspergillosis mycopowder；
Wherein, described PDA solid medium: Rhizoma Solani tuber osi 200g, sucrose 20g, water 1000mL, agar 20g；
Wherein, described aspergillosis solid fermentation culture medium: banana stem leaves slag 85%, cottonseed meal 2%, corn cob 8%, stone powder 1%, thick Semen Maydis
Skin 2%, carbamide 0.4%, dipotassium hydrogen phosphate 0.4%, magnesium sulfate 0.05%, moisture content in medium 50%-60%；The bar of each medium sterilization
Part is: 0.10-0.15MPa, 121 DEG C of sterilizings 30 minutes.
Step 2, the preparation of bacillus amyloliquefaciens mycopowder
Take out bacillus amyloliquefaciens preservation pipe, draw flat board respectively with nutrient solid medium and recover, 30 DEG C of cultivations
48 hours, under flat board, the single colony inoculation of picking was to equipped with nutrient solid medium, cultivates 48 in 30 DEG C of incubators
Hour, with 3000ml sterilized water by the lawn eluting in three Fructus Solani melongenae bottles, it is seeded to equipped with 300L liquid seed culture medium
In 500L fermentation tank, opening stirring 120r/min, within first 10 hours, ventilation is 200L/min, and after 10 hours, ventilation is 320L/min
, cultivate 16-24 hour, treat that total bacterial content is not less than 2,000,000,000 cfu/ml, can be used as bacillus cereus liquid seed for 30 DEG C；
Fermentation: the bacillus cereus liquid seed of above-mentioned preparation is added in spore solid fermentation culture medium, mixing, tray is sent out
Ferment, windrow thickness is 5-10cm, controls fermentation temperature and is 30-40 DEG C, ferments 44 hours, and detection spore content is 15,000,000,000 cfu/
G, can dry, and treats that moisture is less than 10%, i.e. obtains bacillus amyloliquefaciens mycopowder；
Wherein, described nutrient solid medium: peptone 5g, Carnis Bovis seu Bubali cream 3g, sodium chloride 5g, agar 20 g, water 1000mL,
pH7.2；
Wherein, described liquid seed culture medium: glucose 8g/L, Semen Maydis powder 10 g/L, Semen Glycines powder 10 g/L, magnesium sulfate 0.2 g/
L, manganese sulfate 0.1 g/L, potassium dihydrogen phosphate 0.5 g/L, disodium hydrogen phosphate 1.0 g/L, pH7.0；
Wherein, spore solid fermentation culture medium: banana stem leaves slag 60%, wheat bran 35%, cottonseed meal 2%, stone powder 1%, thick skin of Semen Maydis 2%, urine
Element 0.4%, dipotassium hydrogen phosphate 0.4%, magnesium sulfate 0.05%, moisture content in medium 50%-60%；The condition of each medium sterilization is:
0.10-0.15MPa, 121 DEG C of sterilizings 30 minutes.
Step 3, the general preparation receiving streptomycete mycopowder
Take out general streptomyces species preservation pipe of receiving, draw flat board with Gause I solid medium and recover, cultivate 7 days for 30 DEG C,
Under flat board, the streak inoculation of picking single bacterium colony is equipped with in the Fructus Solani melongenae bottle of 150 milliliters of Gause I solid mediums, at incubator
In 30 DEG C cultivate 6-8 days, treat that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore powder and i.e. can use the physiological saline solution of 500 milliliters to wash
De-, regulation spore concentration is 0.1 hundred million cfu/ml, is general streptomycete seed liquor of receiving；
Fermentation: the general streptomycete solid of receiving that streptomycete seed liquor of above-mentioned preparation general being received is seeded to sterilizing with the inoculum concentration of 5% is sent out
In ferment culture medium, strain is received after streptomycete solid medium mixs homogeneously with general, the general streptomycete solid fermentation of receiving that will connect kind
Culture medium is spread out on fermentation tank, and height of materials is 5-8cm, temperature control 28-32 DEG C, and a few days ago air humidity remains 60%-75%, mistake
Rear air humidity remains 40%-50%, ferments 5 days, and detecting its spore content is 1,800,000,000 CFU/g, gets final product low-temperature air-drying, i.e. obtains
General streptomycete mycopowder of receiving；
Wherein, described Gause I solid medium: potassium nitrate: 1 gram, soluble starch: 20 grams, dipotassium hydrogen phosphate: 05
Gram, magnesium sulfate: 05 grams, sodium chloride: 05 grams, ferrous sulfate: 0 01 grams, agar: 20 grams, distilled water supplies 1000ml, PH7
2～7 4；
Wherein, described general streptomycete solid fermentation culture medium of receiving: banana stem leaves slag 80%, cottonseed meal 2%, Semen Maydis powder 1%, stone powder
10%, wheat bran 7%, manganese sulfate 0.004%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate 0.05%, solid medium water content is 40%-60%；
The condition of each medium sterilization is: 0.10-0.15MPa, 121 DEG C of sterilizings 30 minutes.
Step 4 by compound aspergillosis mycopowder, bacillus amyloliquefaciens mycopowder, general streptomycete mycopowder and the cottonseed meal powder received with 1:
The ratio mix homogeneously of 1:1:1, detecting its dirty chromogen hair flat lead fungi spore content is 300,000,000 CFU/g, and lignin peroxidase is lived
It is not less than 55U/ g；Manganese-dependent peroxidase is lived and is not less than 120U/ g；Cellulase content is not less than 150IU/ g, and wood is poly-
Carbohydrase content the lowest 310IU/ g, solves starch spore content and is not less than 3,100,000,000 CFU/g, and general streptomycete content of receiving is not less than 300,000,000
CFU/g, gets final product packed products.
Embodiment 2
Decomposing microbial inoculum application test decomposing microbial inoculum accelerates banana stem leaves residuum and becomes thoroughly decomposed and suppress Fusarium oxysporum disease compliance test result:
Proving ground: large-scale plantation family, Xuwen County Long Tang town；
Arranging following process: process 1. comparisons, soil does not use any decomposing agent, with the fresh water spraying of equivalent at the Fructus Musae pulverized
Stem and leaf residuum；Process 2. and use sale decomposing agent 5kg/ mu on market, dilute 100 times and spray in the banana stem leaves residuum pulverized；
Process 3. and use decomposing agent 2kg/ mu of the present invention, dilute 100 times and spray in the banana stem leaves residuum pulverized.Often process plot area
10 mu.Measuring banana stem leaves tension variations and the change of weight-loss ratio, soil Fusarium oxysporum (easily causes Fructus Musae to wither
Sick) quantity, suppresses the bacillus cereus quantity of Fusarium oxysporum in soil.
Test result indicate that: experiment starts latter 15th day, 35 days and 60 days, compared to control treatment, use the present invention rotten
The process banana stem leaves pulling force of ripe microbial inoculum significantly reduces；And the banana stem leaves pulling force using decomposing microbial inoculum of the present invention process compares city
Sell the pulling force reduction by 25%-56% that decomposing agent processes banana stem leaves.The weight-loss ratio of banana stem leaves measures display, uses the present invention rotten
The banana stem leaves weight-loss ratio that ripe microbial inoculum processes is significantly higher than control treatment, and banana stem leaves weight-loss ratio increases by 2. 1-3. 2 times；This
The banana stem leaves that daylight reason banana stem leaves weight-loss ratio processes also above commercially available decomposing agent.After off-test, soil microorganism measures aobvious
Show, use the present invention and process Fusarium oxysporum quantity in soil and be markedly inferior to control treatment, less than the 1% of control treatment；This
Daylight reason soil Fusarium oxysporum quantity is also markedly inferior to commercially available decomposing agent and processes, only the 10% of control treatment, commercially available straw
Decomposing agent does not has good inhibition to pathogenic soil fungus.The bacillus cereus quantity that can suppress pathogen in soil is surveyed
Fixed display, commercially available straw decomposing inoculant processes and control treatment does not has marked difference, but the present invention processes soil suppression pathogen
Bacillus cereus quantity higher about 1000-10000 times than control treatment.The Population of Actinomycetes suppressing Fusarium oxysporum in soil is relatively executed
Process with conventional decomposing agent and add 100-1000 times.
Claims (3)
1. the banana stem leaves residuum decomposing microbial inoculum suppressing droop, it is characterised in that the Fructus Musae stem of described suppression droop
Leaf residuum decomposing microbial inoculum is mainly by following several bacterium: the dirty flat lead fungi of chromogen hair, Fructus Citri tangerinae green trichoderma, aspergillus oryzae, bacillus amyloliquefaciens,
General streptomycete fermentation of receiving is made；Its concrete preparation method, comprises the following steps:
Step one, the preparation of compound aspergillosis mycopowder
A, the preparation of dirty chromogen hair flat lead fungi seed liquor: take out dirty chromogen hair flat lead fungi culture presevation pipe, use PDA solid medium
Drawing flat board to recover, cultivate 7 days for 30 DEG C, under flat board, the streak inoculation of picking single bacterium colony is equipped with 150 milliliters of PDA solid culture
In the Fructus Solani melongenae bottle of base, cultivate 6-8 days for 30 DEG C in incubator, treat that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore powder and i.e. can use 500
The physiological saline solution eluting of milliliter, regulation spore concentration is 0.01 hundred million cfu/ml, is dirty chromogen hair flat lead fungi seed liquor；
B, the preparation of Fructus Citri tangerinae green trichoderma seed liquor: take out Fructus Citri tangerinae green trichoderma bacterium culture presevation pipe, draw flat board with PDA solid medium and carry out
Recovery, cultivates 6 days for 30 DEG C, and under flat board, the streak inoculation of picking single bacterium colony is equipped with the Fructus Solani melongenae bottle of 150 milliliters of PDA solid mediums
In, cultivate 4-6 days for 30 DEG C in incubator, treat that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore and i.e. can use the aseptic life of 500 milliliters
Reason salt water, regulation spore concentration is 0.1 hundred million cfu/ml, is Fructus Citri tangerinae green trichoderma seed liquor；
C, the preparation of aspergillus oryzae seed liquor: take out aspergillus oryzae strain preservation pipe, draw flat board with PDA solid medium and recover,
Cultivating 5 days for 30 DEG C, under flat board, the streak inoculation of picking single bacterium colony is equipped with in the Fructus Solani melongenae bottle of 150 milliliters of PDA solid mediums,
Incubator is cultivated 5-7 days for 30 DEG C, treats that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore and i.e. can use the sterile physiological salt of 500 milliliters
Water elution, regulation spore concentration is 100,000,000 cfu/ml, is aspergillus oryzae seed liquor；
D, mixed fermentation: by the dirty chromogen hair flat lead fungi seed liquor of above-mentioned preparation, Fructus Citri tangerinae green trichoderma seed liquor and aspergillus oryzae seed liquor
After the ratio mix homogeneously of 2:1:1, being seeded in the aspergillosis solid fermentation culture medium of sterilizing with the inoculum concentration of 2%, strain is with solid
After body culture medium mix homogeneously, being spread out in connecting the aspergillosis solid fermentation culture medium planted on fermentation tank, height of materials is 5-10cm,
Temperature control 28-32 DEG C, humidity remains 60%-75%, ferments 6-10 days, detects its total spore content and is not less than 2,000,000,000 CFU/g, detection
Its dirty chromogen hair flat lead fungi spore content is not less than 1,000,000,000 CFU/g, and lignin peroxidase is lived, i.e. LiP enzyme is lived and is not less than
200U/ g；Manganese-dependent peroxidase, i.e. MnP enzyme is lived and is not less than 500U/ g；Cellulase content is not less than 500IU/ g, wood
Dextranase content the lowest 1000IU/ g, gets final product low-temperature air-drying, and moisture is less than 10%, i.e. obtains compound aspergillosis mycopowder；
Wherein, described PDA solid medium: Rhizoma Solani tuber osi 200g, sucrose 20g, water 1000mL, agar 20g；
Wherein, described aspergillosis solid fermentation culture medium: banana stem leaves slag 85%, cottonseed meal 2%, corn cob 8%, stone powder 1%, thick Semen Maydis
Skin 2%, carbamide 0.4%, dipotassium hydrogen phosphate 0.4%, magnesium sulfate 0.05%, moisture content in medium 50%-60%；The bar of each medium sterilization
Part is: 0.10-0.15MPa, 121 DEG C of sterilizings 30 minutes；
Step 2, the preparation of bacillus amyloliquefaciens mycopowder
Take out bacillus amyloliquefaciens preservation pipe, draw flat board respectively with nutrient solid medium and recover, 30 DEG C of cultivations
48 hours, under flat board, the single colony inoculation of picking was to equipped with nutrient solid medium, cultivates 48 in 30 DEG C of incubators
Hour, with 3000ml sterilized water by the lawn eluting in three Fructus Solani melongenae bottles, it is seeded to equipped with 300L liquid seed culture medium
In 500L fermentation tank, opening stirring 120r/min, within first 10 hours, ventilation is 200L/min, and after 10 hours, ventilation is 320L/min
, cultivate 16-24 hour, treat that total bacterial content is not less than 2,000,000,000 cfu/ml, can be used as bacillus cereus liquid seed for 30 DEG C；
Fermentation: the bacillus cereus liquid seed of above-mentioned preparation is added in spore solid fermentation culture medium, mixing, tray is sent out
Ferment, windrow thickness is 5-10cm, controls fermentation temperature and is 30-40 DEG C, ferments 36-48 hour, treat that spore content is not less than 10,000,000,000
Cfu/g, can dry, and treats that moisture is less than 10%, i.e. obtains bacillus amyloliquefaciens mycopowder；
Wherein, described nutrient solid medium: peptone 5g, Carnis Bovis seu Bubali cream 3g, sodium chloride 5g, agar 20 g, water 1000mL,
pH7.2；
Wherein, described liquid seed culture medium: glucose 8g/L, Semen Maydis powder 10 g/L, Semen Glycines powder 10 g/L, magnesium sulfate 0.2 g/
L, manganese sulfate 0.1 g/L, potassium dihydrogen phosphate 0.5 g/L, disodium hydrogen phosphate 1.0 g/L, pH7.0；
Wherein, spore solid fermentation culture medium: banana stem leaves slag 60%, wheat bran 35%, cottonseed meal 2%, stone powder 1%, thick skin of Semen Maydis 2%, urine
Element 0.4%, dipotassium hydrogen phosphate 0.4%, magnesium sulfate 0.05%, moisture content in medium 50%-60%；The condition of each medium sterilization is:
0.10-0.15MPa, 121 DEG C of sterilizings 30 minutes；
Step 3, the general preparation receiving streptomycete mycopowder
Take out general streptomyces species preservation pipe of receiving, draw flat board with Gause I solid medium and recover, cultivate 7 days for 30 DEG C,
Under flat board, the streak inoculation of picking single bacterium colony is equipped with in the Fructus Solani melongenae bottle of 150 milliliters of Gause I solid mediums, at incubator
In 30 DEG C cultivate 6-8 days, treat that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore powder and i.e. can use the physiological saline solution of 500 milliliters to wash
De-, regulation spore concentration is 0.1 hundred million cfu/ml, is general streptomycete seed liquor of receiving；
Fermentation: the general streptomycete solid of receiving that streptomycete seed liquor of above-mentioned preparation general being received is seeded to sterilizing with the inoculum concentration of 5% is sent out
In ferment culture medium, strain is received after streptomycete solid medium mixs homogeneously with general, the general streptomycete solid fermentation of receiving that will connect kind
Culture medium is spread out on fermentation tank, and height of materials is 5-8cm, temperature control 28-32 DEG C, and a few days ago air humidity remains 60%-75%, mistake
Rear air humidity remains 40%-50%, ferments 4-6 days, detects its spore content and is not less than 1,000,000,000 CFU/g, gets final product low-temperature air-drying,
I.e. obtain general streptomycete mycopowder of receiving；
Wherein, described Gause I solid medium: potassium nitrate: 1 gram, soluble starch: 20 grams, dipotassium hydrogen phosphate: 0.5
Gram, magnesium sulfate: 0.5 gram, sodium chloride: 0.5 gram, ferrous sulfate: 0.01 gram, agar: 20 grams, 1000ml supplied by distilled water,
PH7.2～7.4；
Wherein, described general streptomycete solid fermentation culture medium of receiving: banana stem leaves slag 80%, cottonseed meal 2%, Semen Maydis powder 1%, stone powder
10%, wheat bran 7%, manganese sulfate 0.004%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate 0.05%, solid medium water content is 40%-60%；
The condition of each medium sterilization is: 0.10-0.15MPa, 121 DEG C of sterilizings 30 minutes；
Step 4 by compound aspergillosis mycopowder, bacillus amyloliquefaciens mycopowder, general streptomycete mycopowder and the cottonseed meal powder received with 1:1:1:1
Ratio mix homogeneously, detect its dirty chromogen hair flat lead fungi spore content and be not less than 200,000,000 CFU/g, lignin peroxidase is lived
It is not less than 40U/ g；Manganese-dependent peroxidase is lived and is not less than 100U/ g；Cellulase content is not less than 100IU/ g, and wood is poly-
Carbohydrase content the lowest 200IU/ g, solves starch spore content and is not less than 2,000,000,000 CFU/g, and general streptomycete content of receiving is not less than 200,000,000
CFU/g, gets final product packed products.
A kind of banana stem leaves residuum decomposing microbial inoculum suppressing droop the most according to claim 1, it is characterised in that described
The using method of the banana stem leaves residuum decomposing microbial inoculum of suppression droop is for uniformly spraying 1-5kg decomposing microbial inoculum dilution 100 times
To 1 mu of banana stem leaves residuum.
A kind of banana stem leaves residuum decomposing microbial inoculum suppressing droop the most according to claim 1, it is characterised in that described
The using method of the banana stem leaves residuum decomposing microbial inoculum of suppression droop is every mu and entirely is uniformly spread to by 2-10kg decomposing microbial inoculum
In banana stem leaves residuum.
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| CN113444651A (en) * | 2021-07-12 | 2021-09-28 | 浙江中医药大学 | Saffron endophytic fungus and application thereof in preventing and treating bulb rot |
| CN114574372A (en) * | 2022-03-28 | 2022-06-03 | 山东省林业科学研究院 | Trichoderma citrinoviride and application thereof in degrading fish protein |
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