CN105154336B - A kind of Trichoderma viride XJ 3 and its method for preparing cotton stalk decomposed manure - Google Patents
A kind of Trichoderma viride XJ 3 and its method for preparing cotton stalk decomposed manure Download PDFInfo
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
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- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Abstract
The present invention discloses a kind of Trichoderma viride XJ 3 and its method for preparing cotton stalk decomposed manure.Separation, screening, seed selection, the domestication of strain are carried out by cotton pedotheque, through rDNA ITS sequences measure and Phylogenetic Analysis, it is determined that Trichoderma viride（Trichoderma viride）The CGMCC NO.10989 of XJ 3, and it is inoculated into the cotton stalk after crushing, the length of cotton stalk is 3cm 5cm after wherein crushing, water content is 60% 65%, and add 30% urea, 20% two ammoniums, strain protective agent, by bacterium solution, urea, two ammoniums and strain protective agent are prepared into fermentation base-material, and spray on stalk after being pulverized, plough to cotton and fermented after winter irrigation, the fermentative degradation time of 3 months is up to by autumn to Second Year spring, this prepares the method for cotton stalk decomposed manure in increase output of cotton and increases soil fertility etc. and to have prominent technique effect.
Description
Technical field
The present invention relates to agriculture technical field of microbe application, and specifically, the present invention relates to a kind of voluntarily seed selection to screen
Green wooden enzyme and Trichoderma viride XJ-3 prepare the technical field of cotton stalk decomposed manure.
Background technology
Soil organic matter not only facilitates the sustainable development of agricultural as soil and the important component of land carbon storehouse, and
And the capacity to carbon base in soil, release and turnover etc. all have an impact.Increase soil organic carbon, improve crop yield and water nitrogen
Utilization ratio is the important goal of arid biogeographic zone farmland optimum management.Biological carbon is the organic carbon that a kind of phosphorus content is high, stability is strong,
Soil carbon storage can be effectively increased, is increase soil fertility and productivity.
China is the important cotton production areas in the world, and the cotton stalk after cotton harvesting is a kind of very extensive valuable money of purposes
Source.First, cotton stalk is a kind of good substitute for wood, can be used as producing staple fibre, while can manufacture particieboard, fiber again
The sheet material such as plate and abatvoix, or outclass straw and straw as building materials paper making raw material, quality；Secondly, cotton stalk has very high
Nutritive value, after treatment with micron, its protein content may be up to 12%, higher 5-6 than the protein content of wheat straw and straw
Times, can be as the first-class feed of cattle and sheep；Again, cotton stalk is through the chemical process such as grating and high temperature pyrolysis, produce using carbon monoxide as
Main biogas, a kind of novel energy can be used as；The culture medium of pollution-free food edible mushroom is alternatively arranged as after cotton stalk powder is broken, this
The disposable tableware that external application stalk is made is pollution-free, degradable and renewable.Therefore, for development green agriculture, cotton stalk returns
Abundance and required raw material can be provided after receipts for the deep processing of agricultural.
At present, the general processing method of cotton stalk：First, burning cotton stalk, this is not only a kind of waste, and seriously pollutes
Environment；Second, hand harvest cotton stalk, labor intensity is big, and efficiency is low；Third, direct chopping and returning, though this way can increase soil
The organic fertilizer of earth, but the incidence of disease of coming year cotton diseases and insect pestses can be increased, cotton stalk is not easy to crush, and relies on nature bar merely
Part, cotton stalk corruption ability is weak after crushing, it is difficult to is utilized as organic fertilizer by next batch of cotton.Xinjiang is as the super-huge cotton in China
Flower production base, produces cotton stalk about 5.4 × 10 every year on average6t.Biological carbon returning to the field prepared by cotton stalk pyrolysis, can
To reduce the adverse effect of straw directly returning to field, while it can effectively improve soil productivity.In spite of many on utilizing cotton stalk
Prepare culture medium and research compound fertilizer is used for improving the cotton planting research of soil quality and report, but it is relevant from cotton
In the soil of flower planting site separation screening fermentability by force and the composition such as lignin, cellulose in cotton stalk of can degrading strain,
And the Trichoderma viride for utilizing voluntarily seed selection screening domestication to obtain carries out fermentation cotton stalk and prepares cotton stalk decomposed manure for improveing
The soil on cotton planting ground, the fertility for increasing soil, there is certain preventive and therapeutic effect to cotton in seedling stage damping-off, anthracnose, be favourable
Had not been reported in the research of cotton growth.
The content of the invention
For have no in the prior art about from the soil on cotton planting ground screen Trichoderma viride as excellent species simultaneously
The state of the art of cotton stalk decomposed manure is prepared using the strain of screening, the present invention is intended to provide a kind of Trichoderma viride XJ-3 and its
The method for preparing cotton stalk decomposed manure.The present invention by isolating a collection of microbial strains in the soil on cotton planting ground,
The bacterial strain that one plant of numbering is XJ-3, and the Trichoderma viride for being XJ-3 by using numbering are therefrom separated, addition strain is protected
Protect under conditions of agent, urea and diamines, prepare cotton stalk decomposed manure by raw material of cotton stalk, reduce the unfavorable of straw directly returning to field
Influence, be effectively improved soil productivity, both solved the utilization of cotton stalk discarded object, reduced the pollution for burning cotton stalk to environment, together
When add cotton soil fertility, cotton planting can be improved soil quality, have to cotton in seedling stage damping-off, anthracnose
Certain prevention effect, be advantageous to the plantation of cotton, be advantageous to improve the yield and quality of cotton, and obtain good technology effect
Fruit, the efficient utilization rate of cotton stalk is improved, meaning and the effect in improvement cotton planting ground with reality.
The present invention uses main technical scheme：
By the separation screening of microorganism fungus kind, a collection of microbial strains are isolated in the soil on cotton planting ground, are passed through
Further separation, screening, seed selection, domestication are crossed, obtains the trichoderma viride strain that one plant of numbering is XJ-3.By entering to obtained bacterial strain
Row morphological feature, physio-biochemical characteristics and rDNA ITS sector sequences measure and Phylogenetic Analysis, have primarily determined that it is classified
Status.Meanwhile will number in the cotton stalk that the Trichoderma viride for being XJ-3 is inoculated into after crushing, wherein the length of cotton stalk is after crushing
3cm-5cm, water content 60%-65%, and add 30% urea, 20% two ammoniums, strain protective agent, by bacterium solution, urea,
Two ammoniums and strain protective agent are prepared into fermentation base-material, and spray on stalk after being pulverized, are fermented after winter irrigation of ploughing to cotton,
The fermentative degradation time of 3 months is up to by autumn to Second Year spring, you can cotton stalk decomposed manure is prepared, preparation
Cotton stalk decomposed manure is brown, soft flexible when hand is grabbed；Fermentation system is carried out by using the strain and cotton stalk that voluntarily screen
It is standby to obtain cotton stalk decomposed manure, both solved the utilization of cotton stalk discarded object, reduced the pollution for burning cotton stalk to environment, increased simultaneously
The fertility in cotton soil is added, the quality in soil, is advantageous to the plantation of cotton with can improveing cotton planting, is advantageous to improve cotton
Colored yield and quality, good technique effect is obtained, meaning and the effect in improvement cotton planting ground with reality.
The present invention specifically provides a kind of Trichoderma viride (Trichoderma viride), passes through the soil to cotton planting ground
Middle microorganism is separated, screened and cultivated, and obtains a collection of microbial strains, therefrom filters out one plant of Serial No. XJ-3 bacterium
Strain, through microbiological classification and identification, the bacterial strain belongs to Trichoderma viride.
Specifically, the present invention provides a kind of Trichoderma viride (Trichoderma viride), strain number XJ-3.The bacterium
Strain was preserved in budapest treaty microorganism International Depository Authority before the applying date：Chinese microorganism strain preservation conservator
Can common micro-organisms center (CGMCC).Address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Chinese Academy of Sciences microorganism are ground
Study carefully institute, postcode：100101.Preservation date is on June 23rd, 2015, and preserving number is CGMCC No.10989.Reflected through microbiology
It is set to Trichoderma viride.The bacterial strain optimal culture conditions：25 DEG C -30 DEG C, pH 5-5.5, culture medium Ma Ling of temperature
Potato agar glucose (PDA) culture medium；Be white when the bacterial strain bacterium colony starts, it is fine and close, it is circular, extended to surrounding, after from bacterium colony
Center produces green spores, and center becomes green, and periphery of bacterial colonies has the growth band of white hypha, and last whole bacterium colony all becomes
Green；Spore is in ellipse, CO2Influence to the strain growth depends on CO2Concentration and culture medium pH value, in alkaline matrix
In, the CO of high concentration2Be advantageous to the growth that numbering is XJ-3 bacterial strains；The bacterial strain can by the use of gas chromatography as carbon source, compared with
Preferably monosaccharide and disaccharide, polysaccharide, purine, pyrimidine and amino acid etc.；The bacterial strain is big on the culture medium rich in carbohydrate
Amount produces acids, and 60%-80% citric acid is produced as carbon source, the bacterium by the use of glucose or starch；In buffer medium, ammonium
It is the nitrogen source that the bacterial strain most easily utilizes, other nitrogen sources such as amino acid, urea, nitrate, nitrite can also maintain it normally to give birth to
It is long；Grown with asparagus fern Men Suwei nitrogen sources especially good, nitrogen content is low, can promote sporogenesis, to the negatively influencing of high concentration nitrate,
Because the presence of magnesium sulfate can be compensated；The growth of inorganic salts and micro- inorganic salts to the bacterial strain is critically important, magnesium from
Son can promote its growth, and copper ion can promote conidium pigment to be formed, and formation of the iron ion to spore plays an important roll, warp
Microbiology is accredited as Trichoderma viride bacterium, but has obvious difference with common Trichoderma viride strain.Reference
《Fungal identification handbook》System Physiology and biochemistry identification is carried out to numbering XJ-3 bacterial strains, is detected through Physiology and biochemistry and determines numbering XJ-3 bacterium
Strain is the member in trichoderma (Trichoderma).Pass through the homologous comparisons of BLAST, strain X J-3 ITS rDNA sequence nucleotide sequences
After carrying out BLAST analyses in ncbi database, constructing system chadogram, the numbering is strain X J-3 and Trichoderma
Longibrachiatum isolateT50 are in minimum branch, are its allied specieses, and its homology is 76%；And then by the bacterial strain
XJ-3 is defined as Trichoderma viride, identifies and classifies by said system taxology, it was demonstrated that be provided by the invention green
Color trichoderma (Trichoderma viride) has with common Trichoderma viride in physio-biochemical characteristics and molecular level etc.
Obvious otherness, according to strain Analysis of The Physiological And Biochemical Properties, the comprehensive identification of molecular level analysis and systematics, from point
It is Trichoderma viride (Trichoderma viride) that the bacterial strain is identified in class.
Meanwhile the specific one kind that provides of the present invention utilizes Trichoderma viride (Trichoderma viride) XJ-3 CGMCC
The method that NO.10989 prepares cotton stalk decomposed manure, specific preparation method step are as follows：
(1) strain activation and culture：Prepare Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989 bacterium
The potato dextrose agar of strain, sterile working is strictly carried out after sterilizing, flat board is forwarded to from inclined-plane, at 28 DEG C of temperature
Culture 5 days.
(2) one-level culture：It is forwarded to from picking single bacterium colony on the potato dextrose agar solid medium in step (1)
In 50mL conical flasks equipped with potato dextrose broth, sterile working, 28 DEG C, 120r/min cultures 24h-48h.
(3) two level expands culture：Above-mentioned steps one-level culture strain is inoculated in and fills potato glucose Liquid Culture
In the 500mL conical flasks of base, sterile working, 28 DEG C, 120r/min cultures 24h-48h.
(4) determination of water：It is 250 kilograms to 450 kilograms of ratio core according to rear every mu of cotton stalk yield of cotton is plucked
Meter standard, cotton stalk processing need to add the 60%-65% that water is cotton stalk yield.
(5) it is inoculated with：Trichoderma viride (Trichoderma viride) XJ-3 CGMCC prepared by step (3)
In water prepared by NO.10989 activated spawns inoculation above-mentioned steps (4), prepared by water 10%-15% ratio inoculation as required
Trichoderma viride microbial inoculum is obtained, and strain protective agent, urea and diamines are added respectively according to the microbial inoculum amount of preparation, wherein, strain is protected
Agent is protected by sucrose, sodium glutamate and glycerine according to weight ratio 1:1:1 proportioning, strain protective agent is according to 15% addition, urea and two
Amine is added according to 30% and 20% ratio respectively, and fermentation base-material is prepared by stirring.
(6) cotton stalk crushing returning to the field：After having plucked cotton in autumn, by crop field operation by cotton stalk crushing, cotton stalk after crushing
Length control be 3cm-5cm, the hair prepared in crushing process using step (5) is sprayed on spray equipment cotton stalk after being pulverized
Ferment base-material.
(7) ferment：Fermentation base-material prepared by step (6) is raised together in company with the cotton stalk after crushing and is sprinkling upon cotton field
In, buried using common equipment of the ploughing base-material that will ferment, irrigate using winter irrigation water by cotton field flood is climing, autumn last year to the
Cotton stalk is prepared into cotton stalk decomposed manure by fermenting during 2 year spring.
Further, the present invention provides utilizes Trichoderma viride (Trichoderma viride) XJ-3 CGMCC using above-mentioned
The cotton stalk decomposed manure that the method that NO.10989 prepares cotton stalk decomposed manure prepares.
The present invention is provided one kind and prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
The method of cotton stalk decomposed manure, add urea and diamines, one side Trichoderma viride (Trichoderma viride) XJ-3
CGMCC NO.10989 bacterial strains need the carbon source and nitrogen source of abundance in its growth metabolism, add urea and diamines is advantageous to strain
The growth and breeding of Trichoderma viride；On the other hand addition urea and diamines can promote the fermentation of cotton stalk, while also help and change
The quality on kind cotton planting ground and the growth for promoting next batch of cotton.
The present invention is provided one kind and prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
The method of cotton stalk decomposed manure, strain protective agent is added, be on the one hand advantageous to protect Trichoderma viride (Trichoderma
Viride) the normal growth metabolism of XJ-3 CGMCC NO.10989 bacterial strains；On the other hand nutrients is provided for the growth of bacterial strain
Matter, be advantageous to the fermentation of cotton stalk.The strain protective agent of addition is by sucrose, sodium glutamate and glycerine according to weight ratio 1:1:1
Proportioning, adding sucrose and sodium glutamate contributes to strain to provide certain sugar source and energy element in fermented and cultured in cotton field
Element, help the breeding of strain, while can also cause strain to obtain certain protection, using glycerine ensure strain big Tanaka with
A kind of temperature low safeguard measure when the cotton stalk of crushing is survived the winter in cotton field, can promote the fermentation of cotton stalk, simultaneously
Also help the quality for improving cotton planting ground and the growth for promoting next batch of cotton.
In the present invention, potato glucose (PDA) Solid agar culture, the potato dextrose broth of use
All it is common culture medium, its culture medium content and component, and the condition of culture of culture medium, those of ordinary skill in the art can
With by that can be looked into known technology reference book or document.
By implementing the specific content of the invention of the present invention, following beneficial effect can be reached:
(1) Trichoderma viride (Trichoderma viride) the XJ-3 CGMCC NO.10989 bacterial strains tool that the present invention screens
There is stronger growth and breeding ability, growth rate is fast, and hereditary capacity is stable, and has specific effect to degraded cotton stalk.
(2) present invention is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989 bacterial strains
Cotton stalk decomposed manure, it is brown, it is soft flexible when hand is grabbed.
(3) present invention is a kind of is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
The method of cotton stalk decomposed manure, both solved the utilization of cotton stalk discarded object, reduced the pollution for burning cotton stalk to environment, increased simultaneously
The fertility in cotton soil is added, the quality in soil, is advantageous to the plantation of cotton with can improveing cotton planting, is advantageous to improve cotton
Colored yield and quality, good technique effect is obtained, meaning and the effect in improvement cotton planting ground with reality.
(4) present invention is a kind of is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
The method of cotton stalk decomposed manure, the adverse effect of straw directly returning to field is reduced, be effectively improved soil productivity, while improve cotton
The utilization rate of bar.
(5) present invention is a kind of is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
The method of cotton stalk decomposed manure produces beneficial effect to soil fertility.Cotton stalk decomposed manure group available potassium prepared by the present invention
Content organizes raising 44.57% with being directly used in cotton than cotton stalk, and available phosphorus contents organize raising with being directly used in cotton compared with cotton stalk
136.86%, water-soluble calcium content organizes raising 215.38% with being directly used in cotton than cotton stalk, while can promote organic in soil
Matter is decomposed, and the content of organic matter is organized with being directly used in cotton compared with cotton stalk reduces by 14.06% respectively.
(6) present invention is a kind of is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
The method of cotton stalk decomposed manure can increase the yield of cotton, and seed cotton yield organizes volume increase 21.20% with being directly used in cotton than cotton stalk,
Than cotton stalk, group adds 0.16g to unginned cotton's bell with being directly used in cotton again, and single basal munure is 15.6, and height is organized with being directly used in cotton compared with cotton stalk
2.40, therefore, cotton stalk decomposed manure group prepared by the present invention has certain production-increasing function to cotton.
(7) present invention is a kind of is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
The method of cotton stalk decomposed manure has certain prevention effect to cotton in seedling stage damping-off, anthracnose.Wherein, prepared by the present invention
The cotton stalk decomposed manure group incidence of disease minimum 0.67%, the diseased plant rate of commercially available common organic fertilizer group is 1.33%, and cotton stalk is direct
The diseased plant rate highest for cotton organized, it is 2.01%.
Brief description of the drawings
The shown in green trichodermas of Fig. 1 (Trichoderma viride) XJ-3 CGMCC NO.10989 colonial morphology figure,
A is bacterium colony front in figure, and B is the bacterium colony back side.
The shown in green trichodermas of Fig. 2 (Trichoderma viride) the XJ-3 CGMCC NO.10989 micro- knot of spore
Composition.
The shown in green trichodermas of Fig. 3 (Trichoderma viride) the XJ-3 CGMCC NO.10989 micro- knot of mycelia
Composition.
The shown in green trichodermas of Fig. 4 (Trichoderma viride) XJ-3 CGMCC NO.10989 rDNA ITS areas
Section sequential system development tree graph.
Embodiment
Below, the present invention is illustrated for embodiment, still, the present invention is not limited to following embodiments.
All raw and auxiliary materials selected in the present invention, and bacterium culture medium, the cultural method selected are all ripe for this area
Know selection, the % being related in the present invention is weight percentage, unless otherwise indicated except.
Embodiment one：Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989 separation, screening and
Identification
1st, the separation and screening of strain
(1) separate
Trichoderma viride used in the present invention is sampled from the soil of Xinjiang South Sinkiang Cotton Cultivation in Akesu Region planting site
Separation, according to strain degradation Analysis on action mechanism, preliminary screening goes out to have cotton stalk the bacterial strain of certain degradation capability.Utilize
Traditional plating method isolates the microorganism in soil layer, plate streak purifying bacterial strain, with different cultivation temperatures, pH
Value, culture medium are enrichment condition, filter out a collection of well-grown microbial strains, therefrom preferably go out one plant of Serial No. XJ-3
Bacterial strain.
Separating step：According to gradient dilution method, pedotheque is weighed in 10g cotton fields in 90mL sterile salines, 30
Gradient dilution is carried out after DEG C activation 30min, chooses 10-2、10-3、10-4Dilution is respectively coated on potato dextrose agar
(PDA) flat board of culture medium, 3 repetitions are each handled, in 28 DEG C of cultures.Picking shape, size, color etc. after bacterium colony is grown
Different bacterium colony difference streak inoculations are in new isolation medium potato dextrose agar (PDA) culture medium, until without miscellaneous bacteria
Fall.A bacterial strain part after purification is stored in 4 using mode preservation, a parts such as lyophilized products ampoul tube, glycerol tube and liquid nitrogen
DEG C it is directly used in follow-up study.
(2) condition of culture
By in the inoculation of purifying to potato dextrose agar (PDA) culture medium slant, cultivated in 28 DEG C
48h, it is put into 4 DEG C of refrigerators and saves backup.
Growth temperature：Numbering is that the growth temperature range of XJ-3 bacterial strains is 20 DEG C -38 DEG C, and optimum temperature is 25 DEG C -30 DEG C.
Grow pH：Numbering is that can be grown on the culture medium that the growth pH value of XJ-3 bacterial strains is 1.5 or 9.0, but acid bar
Part is higher than the germination rate under alkalescence condition, and growth pH scopes are 1.5-9.0, and the most suitable growth pH value is 5-5.5.
Carbon source：Numbering is that XJ-3 bacterial strains can be by the use of gas chromatography as carbon source, preferably monosaccharide and disaccharide, more
Sugar, purine, pyrimidine and amino acid etc..
Nitrogen source：In buffer medium, ammonium is that numbering is nitrogen source that XJ-3 bacterial strains most easily utilize, other nitrogen sources such as amino acid,
Urea, nitrate, nitrite can maintain its normal growth；When wherein with asparagus fern Men Suwei nitrogen sources, strain growth is good, nitrogenous
Measure low, can promote sporogenesis, magnesium sulfate can reduce high concentration nitrate to negatively influencing that numbering is XJ-3 bacterial strains.
Inorganic salts and trace element：It is XJ-3 strain growths that magnesium ion, which can promote numbering, and it is XJ- that copper ion, which can promote numbering,
3 bacterial strain conidium pigments are formed, and formation of the iron ion to spore plays an important roll.
Potato dextrose agar (PDA) culture medium is prepared as：Potato is cleaned peeling, takes 200 grams to be cut into
Fritter, add 1000 milliliters of water, after boiling half an hour, supply moisture.10 grams of agar are added in filtrate, add sucrose after boiling dissolving
20 grams, moisture is supplied, is dispensed, sterilizing, you can prepare PDA solid mediums.
By the way that the Soil Microorganism on cotton planting ground is separated, screened and cultivated, a collection of microbial strains are obtained,
One plant of Serial No. XJ-3 bacterial strain is therefrom filtered out, through microbiological classification and identification, the bacterial strain belongs to Trichoderma
viride.The present invention provides a kind of Trichoderma viride (Trichoderma viride), strain number XJ-3.The bacterial strain is in Shen
Budapest treaty microorganism International Depository Authority is please preserved in a few days ago：China Committee for Culture Collection of Microorganisms is commonly micro-
Bio-Centers (CGMCC).Address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postal
Compile：100101.Preservation date is on June 23rd, 2015, and preserving number is CGMCC No.10989.It is accredited as through microbiology
Trichoderma viride.The bacterial strain optimal culture conditions：25 DEG C -30 DEG C of temperature, pH 5-5.5, culture medium is potato
Agar glucose (PDA) culture medium；Be white when the bacterial strain bacterium colony starts, it is fine and close, it is circular, extended to surrounding, after from bacterium colony
Centre produces green spores, and center becomes green, and periphery of bacterial colonies has the growth band of white hypha, and last whole bacterium colony all becomes green
Color；Spore is in ellipse, CO2Influence to the strain growth depends on CO2Concentration and culture medium pH value, in alkaline matrix,
The CO of high concentration2Be advantageous to the growth that numbering is XJ-3 bacterial strains；The bacterial strain can be more satisfactory by the use of gas chromatography as carbon source
Be monosaccharide and disaccharide, polysaccharide, purine, pyrimidine and amino acid etc.；The bacterial strain produces greatly on the culture medium rich in carbohydrate
Acid is measured, 60%-80% citric acid is produced as carbon source, the bacterium by the use of glucose or starch；In buffer medium, ammonium
It is the nitrogen source that the bacterial strain most easily utilizes, other nitrogen sources such as amino acid, urea, nitrate, nitrite can also maintain it normally to give birth to
It is long；During with asparagus fern Men Suwei nitrogen sources, growth is especially good, and nitrogen content is low, can promote sporogenesis；The presence of magnesium sulfate can reduce
High concentration nitrate is to the negatively influencing of the bacterial strain, due to being compensated；The life of inorganic salts and micro- inorganic salts to the bacterial strain
Length is critically important, and magnesium ion can promote its growth, and copper ion can promote conidium pigment to be formed, formation tool of the iron ion to spore
Play an important role, be accredited as Trichoderma viride bacterium through microbiology, but have with common Trichoderma viride strain bright
The difference of aobvious colonial morphology, Physiology and biochemistry.Reference《Fungal identification handbook》System Physiology and biochemistry is carried out to numbering XJ-3 bacterial strains
Identification, detected through Physiology and biochemistry and determine that numbering XJ-3 bacterial strains are the member in trichoderma (Trichoderma).It is same by BLAST
Source compares, after strain X J-3 ITS rDNA sequence nucleotide sequences carry out BLAST analyses in ncbi database, constructing system evolution
Tree, the numbering are that strain X J-3 and Trichoderma longibrachiatum isolateT50 are in minimum branch, are them
Allied species, its homology are 76%；And then strain X J-3 is defined as Trichoderma viride, by said system point
Class identification classification, it was demonstrated that Trichoderma viride provided by the invention (Trichoderma viride) has with common Trichoderma viride
The otherness of obvious physio-biochemical characteristics difference and molecular level, according to strain Analysis of The Physiological And Biochemical Properties, molecular water is divided equally
Analysis and the comprehensive identification of systematics, the strain that numbering is XJ-3 is although in terms of physio-biochemical characteristics and molecular level and often
The Trichoderma viride seen has obvious difference, is a kind of new bacterium, but from taxology, is accredited as Trichoderma viride (Trichoderma
Viride), referring to accompanying drawing 1,2 and accompanying drawing 3.
Embodiment two：Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989 separation, screening and
Identification
1st, PCR expands rDNA ITS sections and its sequencing of Trichoderma viride
The single bacterium colony of a small amount of XJ-3 bacterial strains of picking, it is put into the EP pipes for filling 25 μ L sterilized waters, 100 DEG C are boiled 8-10min,
It is put into 5min in mixture of ice and water rapidly afterwards.10000r/min, 5min, 4 DEG C of preservations are centrifuged, the used time takes supernatant.
The structure of rDNA ITS sections gene sequencings and its systematic evolution tree：The DNA of bacterial strain is extracted according to a conventional method,
Universal primer will be diluted with deionized water, performing PCR of going forward side by side expands, and design of primers is as follows:
ITS1(F)：5'-TCCGTAGGTGAACCTGCGG-3'
ITS4(R)：5'-TCCTCCGCTTATTGATATGC-3'
The structure of rDNA ITS sections gene sequencings and its systematic evolution tree：The total of bacterial strain is extracted according to a conventional method
DNA, the PCR for diluting universal primer rDNA ITS sections is expanded with deionized water.Primer passes through through synthesis, electrophoresis detection
Sequencing.
2nd, the comparison of rDNA ITS sections gene order and Phylogenetic Analysis
Obtained rDNA ITS sections gene orders and the nucleotide sequence progress in GenBank databases will be sequenced
BLAST is analyzed, and therefrom rDNA ITS section gene orders similar in acquisition, XJ-3 rDNA ITS section gene orders are existed
After BLAST analyses being carried out in ncbi database, constructing system chadogram.Referring to shown in accompanying drawing 4, strain X J-3 with
Evolutionary distance is most short between Trichoderma longibrachiatum isolateT50, is Trichoderma
Longibrachiatum isolate allied species.With reference to XJ-3 Morphologic Characteristics and physio-biochemical characteristics, determine that it is
Trichoderma viride (Trichoderma viride) belongs to.Measured rDNA ITS sections gene order is inputted into Genbank, with
Blast programs carry out tetraploid rice, find the rDNA of it and Trichoderma longibrachiatum isolateT50
The similitude of ITS section gene orders is maximum, similitude 76%, so as to be further determined as Trichoderma viride,
There is obvious otherness in physio-biochemical characteristics difference and molecular level etc. with common Trichoderma viride, be a kind of typical case
New bacterium.With reference to XJ-3 Morphologic Characteristics and physio-biochemical characteristics, from classification evolution angle-determining, it for bacterium numbering is
XJ-3 comprehensive identifications are Trichoderma viride (Trichoderma viride)
Embodiment three：Cotton stalk is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
Decomposed manure
Cotton stalk decomposed manure is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
Specific preparation method step it is as follows：
(1) strain activation and culture：Prepare Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989 bacterium
The potato dextrose agar of strain, sterile working is strictly carried out after sterilizing, flat board is forwarded to from inclined-plane, at 28 DEG C of temperature
Culture 5 days.
(2) one-level culture：It is forwarded to from picking single bacterium colony on the potato dextrose agar solid medium in step (1)
In 50mL conical flasks equipped with potato dextrose broth, sterile working, 28 DEG C, 120r/min cultures 24h-48h.
(3) two level expands culture：Above-mentioned steps one-level culture strain is inoculated in and fills potato glucose Liquid Culture
In the 500mL conical flasks of base, sterile working, 28 DEG C, 120r/min cultures 24h-48h.
(4) determination of water：It is 250 kilograms to 450 kilograms of ratio core according to rear every mu of cotton stalk yield of cotton is plucked
Meter standard, cotton stalk processing need to add the 60%-65% that water is cotton stalk yield.
(5) it is inoculated with：Trichoderma viride (Trichoderma viride) XJ-3 CGMCC prepared by step (3)
In water prepared by NO.10989 activated spawns inoculation above-mentioned steps (4), prepared by water 10%-15% ratio inoculation as required
Trichoderma viride microbial inoculum is obtained, and strain protective agent, urea and diamines are added respectively according to the microbial inoculum amount of preparation, wherein, strain is protected
Agent is protected by sucrose, sodium glutamate and glycerine according to weight ratio 1:1:1 proportioning, strain protective agent is according to 15% addition, urea and two
Amine is added according to 30% and 20% ratio respectively, and fermentation base-material is prepared by stirring.
(6) cotton stalk crushing returning to the field：After having plucked cotton in autumn, by crop field operation by cotton stalk crushing, cotton stalk after crushing
Length control be 3cm-5cm, the hair prepared in crushing process using step (5) is sprayed on spray equipment cotton stalk after being pulverized
Ferment base-material.
(7) ferment：Fermentation base-material prepared by step (6) is raised together in company with the cotton stalk after crushing and is sprinkling upon cotton field
In, buried using common equipment of the ploughing base-material that will ferment, irrigate using winter irrigation water by cotton field flood is climing, autumn last year to the
Cotton stalk is prepared into cotton stalk decomposed manure by fermenting during 2 year spring.
In the present invention, using the cotton of Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989 preparations
Brown is presented in bar decomposed manure, soft flexible when hand is grabbed.The present invention is carried out by using the strain and cotton stalk voluntarily screened
Fermentation prepares cotton stalk decomposed manure, has both solved the utilization of cotton stalk discarded object, has reduced the pollution for burning cotton stalk to environment,
The fertility in cotton soil is added simultaneously, is advantageous to the plantation of cotton.
Example IV：Cotton stalk is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
Decomposed manure
Cotton stalk decomposed manure is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
Specific preparation method step it is as follows：
(1) strain activation and culture：Prepare Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989 bacterium
The potato dextrose agar of strain, sterile working is strictly carried out after sterilizing, flat board is forwarded to from inclined-plane, at 28 DEG C of temperature
Culture 5 days.
(2) one-level culture：It is forwarded to from picking single bacterium colony on the potato dextrose agar solid medium in step (1)
In 50mL conical flasks equipped with potato dextrose broth, sterile working, 28 DEG C, 120r/min cultures 24h-48h.
(3) two level expands culture：Above-mentioned steps one-level culture strain is inoculated in and fills potato glucose Liquid Culture
In the 500mL conical flasks of base, sterile working, 28 DEG C, 120r/min cultures 24h-48h.
(4) determination of water：Standard, cotton are assessed according to the ratio that rear every mu of cotton stalk yield of cotton is 250 kilograms is plucked
The processing of chopped straw stalk needs to add 60% that water is cotton stalk yield, therefore water is 150L.
(5) it is inoculated with：Trichoderma viride (Trichoderma viride) XJ-3 CGMCC prepared by step (3)
In water prepared by NO.10989 activated spawns inoculation above-mentioned steps (4), the ratio inoculation of water 10% as required prepares
Trichoderma viride microbial inoculum, the bacterium solution of access is 15L, and adds strain protective agent, urea and diamines respectively according to the microbial inoculum amount of preparation,
Wherein, strain protective agent by sucrose, sodium glutamate and glycerine according to weight ratio 1:1:1 proportioning, strain protective agent add according to 15%
Add, addition is 24.75 kilograms, and urea and diamines add according to 30% and 20% ratio respectively, and addition is 49.5 kilograms
With 53 kilograms, and stir and prepare fermentation base-material.
(6) cotton stalk crushing returning to the field：After having plucked cotton in autumn, by crop field operation by cotton stalk crushing, cotton stalk after crushing
Length control is 3cm-5cm, will spray the hair of step (5) preparation on the cotton stalk after crushing using spray equipment in crushing process
Ferment base-material.
(7) ferment：Fermentation base-material prepared by step (6) is raised together in company with the cotton stalk after crushing and is sprinkling upon cotton field
In, buried using common equipment of the ploughing base-material that will ferment, irrigate using winter irrigation water by cotton field flood is climing, autumn last year to the
Cotton stalk is prepared into cotton stalk decomposed manure by fermenting during 2 year spring.
The present invention adds strain protective agent when fermentation prepares cotton stalk decomposed manure, is on the one hand advantageous to protection green
The normal growth metabolism of trichoderma (Trichoderma viride) XJ-3 CGMCC NO.10989 bacterial strains；On the other hand it is bacterial strain
Growth provide nutriment, be advantageous to the fermentation of cotton stalk.The strain protective agent of addition is by sucrose, sodium glutamate and glycerine
According to weight than 1:1:1 proportioning, adding sucrose and sodium glutamate contributes to strain to be provided necessarily in fermented and cultured in cotton field
Sugar source and energy element, help the breeding of strain, while can also cause strain to obtain certain protection, bacterium ensured using glycerine
A kind of kind low safeguard measure of temperature when big Tanaka survives the winter with the cotton stalk of crushing in cotton field, can promote cotton
The fermentation of bar, while also help the quality for improving cotton planting ground and promote the growth of next batch of cotton.
Embodiment five：Cotton stalk is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
Decomposed manure
Cotton stalk decomposed manure is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
Specific preparation method step it is as follows：
(1) strain activation and culture：Prepare Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989 bacterium
The potato dextrose agar of strain, sterile working is strictly carried out after sterilizing, flat board is forwarded to from inclined-plane, at 28 DEG C of temperature
Culture 5 days.
(2) one-level culture：It is forwarded to from picking single bacterium colony on the potato dextrose agar solid medium in step (1)
In 50mL conical flasks equipped with potato dextrose broth, sterile working, 28 DEG C, 120r/min cultures 24h-48h.
(3) two level expands culture：Above-mentioned steps one-level culture strain is inoculated in and fills potato glucose Liquid Culture
In the 500mL conical flasks of base, sterile working, 28 DEG C, 120r/min cultures 24h-48h.
(4) determination of water：Standard, cotton are assessed according to the ratio that rear every mu of cotton stalk yield of cotton is 450 kilograms is plucked
The processing of chopped straw stalk needs to add 65% that water is cotton stalk yield, therefore water is 292.5L.
(5) it is inoculated with：Trichoderma viride (Trichoderma viride) XJ-3 CGMCC prepared by step (3)
In water prepared by NO.10989 activated spawns inoculation above-mentioned steps (4), the ratio inoculation of water 15% as required prepares
Trichoderma viride microbial inoculum, the bacterium solution of access is 43.88L, and adds strain protective agent, urea and two respectively according to the microbial inoculum amount of preparation
Amine, wherein, strain protective agent is by sucrose, sodium glutamate and glycerine according to weight ratio 1:1:1 proportioning, strain protective agent is according to 15%
Addition, addition are 50.46 kilograms, and urea and diamines add according to 30% and 20% ratio respectively, addition 100.91
Kilogram and 67.28 kilograms, and stir and prepare fermentation base-material.
(6) cotton stalk crushing returning to the field：After having plucked cotton in autumn, by crop field operation by cotton stalk crushing, cotton stalk after crushing
Length control is 3cm-5cm, will spray the hair of step (5) preparation on the cotton stalk after crushing using spray equipment in crushing process
Ferment base-material.
(7) ferment：Fermentation base-material prepared by step (6) is raised together in company with the cotton stalk after crushing and is sprinkling upon cotton field
In, buried using common equipment of the ploughing base-material that will ferment, irrigate using winter irrigation water by cotton field flood is climing, autumn last year to the
Cotton stalk is prepared into cotton stalk decomposed manure by fermenting during 2 year spring.
Embodiment six：Cotton stalk is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
Decomposed manure
Cotton stalk decomposed manure is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
Specific preparation method step it is as follows：
(1) strain activation and culture：Prepare Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989 bacterium
The potato dextrose agar of strain, sterile working is strictly carried out after sterilizing, flat board is forwarded to from inclined-plane, at 28 DEG C of temperature
Culture 5 days.
(2) one-level culture：It is forwarded to from picking single bacterium colony on the potato dextrose agar solid medium in step (1)
In 50mL conical flasks equipped with potato dextrose broth, sterile working, 28 DEG C, 120r/min cultures 24h-48h.
(3) two level expands culture：Above-mentioned steps one-level culture strain is inoculated in and fills potato glucose Liquid Culture
In the 500mL conical flasks of base, sterile working, 28 DEG C, 120r/min cultures 24h-48h.
(4) determination of water：Standard, cotton are assessed according to the ratio that rear every mu of cotton stalk yield of cotton is 350 kilograms is plucked
The processing of chopped straw stalk needs to add 62% that water is cotton stalk yield, therefore water is 217L.
(5) it is inoculated with：Trichoderma viride (Trichoderma viride) XJ-3 CGMCC prepared by step (3)
In water prepared by NO.10989 activated spawns inoculation above-mentioned steps (4), the ratio inoculation of water 12% as required prepares
Trichoderma viride microbial inoculum, the bacterium solution of access is 26.04L, and adds strain protective agent, urea and two respectively according to the microbial inoculum amount of preparation
Amine, wherein, strain protective agent is by sucrose, sodium glutamate and glycerine according to weight ratio 1:1:1 proportioning, strain protective agent is according to 15%
Addition, addition are 36.46 kilograms, and urea and diamines add according to 30% and 20% ratio respectively, and addition is 72.91 public
Jin and 48.61 kilograms, and stir and prepare fermentation base-material.
(6) cotton stalk crushing returning to the field：After having plucked cotton in autumn, by crop field operation by cotton stalk crushing, cotton stalk after crushing
Length control is 3cm-5cm, will spray the hair of step (5) preparation on the cotton stalk after crushing using spray equipment in crushing process
Ferment base-material.
(7) ferment：Fermentation base-material prepared by step (6) is raised together in company with the cotton stalk after crushing and is sprinkling upon cotton field
In, buried using common equipment of the ploughing base-material that will ferment, irrigate using winter irrigation water by cotton field flood is climing, autumn last year to the
Cotton stalk is prepared into cotton stalk decomposed manure by fermenting during 2 year spring.
Embodiment seven：Cotton stalk decomposed manure is advantageous to the field experiment of cotton growth
1st, experimental design and processing
This experiment is carried out in the cotton planting ground of Aksu Prefecture.Before this research, continuously planted for 2 seasons to cotton
Cotton, wherein earth type plough desert grey soil to fill, and quality is attached most importance to earth, organic matter 9.42g/kg, alkali-hydrolyzable nitrogen 36.76mg/kg, full nitrogen
0.19g/kg, rapid available phosphorus 27.42mg/kg, available potassium 284.82mg/kg.
Experiment shares 2 processing and 1 control, i.e.,：Cotton stalk with being directly used in cotton group, commercially available common organic fertilizer group, this hair
It is bright to prepare cotton stalk decomposed manure group using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989.Cotton
The seeds of flowering plants, which is planted, uses plastic-film-covered cultivation, the row of a film 4, Spacing form 30cm+60cm+30cm, spacing in the rows 10cm, and thickness of sowing 22.2 ×
104Strain/hm2.Irrigation method is under-film drip irrigation, and a film two is managed, drip irrigation zone spacing 90cm.Using it is dry broadcast it is wet go out, after planting ooze seedling
Water 45mm.Poured water during cotton growth 9 times, total irrigation volume 600mm, other cultivation management measures are with reference to local crop field.
2nd, process is tested
In the different growth phases locality upper plant sample of cotton, separated according to stem, leaf, cotton buds and bolls Different Organs, in 105
At DEG C after fixing 30min, then constant weight is dried at 70 DEG C, finally weighed.
Cotton disease is investigated in cotton in seedling stage；Cell is divided to receive flower meter production, measure bell weight；The fertility of soil.
3rd, result of the test
(1) dry-matter accumulation changes
Overall apparently cotton dry matter content extends with cotton growing stage, cotton stalk with being directly used in cotton group, commercially available common
Organic fertilizer group, the present invention prepare cotton stalk corruption using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
Difference between ripe organic fertilizer group constantly increases.It is flat that cotton stalk organizes cotton dry-matter accumulation increase within growth period with being directly used in cotton
It is slow；But commercially available common organic fertilizer group and the present invention utilize Trichoderma viride (Trichoderma viride) XJ-3 CGMCC
NO.10989 prepares cotton stalk decomposed manure group dry-matter accumulation after cotton florescence and increased sharply, gradually flat after Shengjing Town
It is slow.Meanwhile using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989 to prepare cotton stalk decomposed by the present invention
Dry matter accumulation amount of the dry matter accumulation amount of organic fertilizer group cotton higher than commercially available common organic fertilizer group cotton.
(2) the shadow noon of disease in cotton seedling stage
Sprout term disease is mainly anthracnose and damping-off.The present invention utilizes Trichoderma viride (Trichoderma viride)
XJ-3 CGMCC NO.10989 prepare cotton stalk decomposed manure group the incidence of disease be below cotton stalk organize with being directly used in cotton with it is commercially available
Common organic fertilizer group, wherein the present invention utilizes Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989 systems
The standby cotton stalk decomposed manure group incidence of disease minimum 0.67%, the diseased plant rate of commercially available common organic fertilizer group is 1.33%, and cotton stalk is straight
The diseased plant rate highest for cotton organized is connect, is 2.01%.As a result show that the present invention utilizes Trichoderma viride (Trichoderma
Viride) XJ-3 CGMCC NO.10989 prepare cotton stalk decomposed manure group have to cotton in seedling stage damping-off, anthracnose it is certain
Prevention effect.
(3) present invention prepares cotton stalk using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
Influence of the decomposed manure to output of cotton constituent element
It is decomposed that the present invention using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989 prepares cotton stalk
Organic fertilizer group seed cotton yield organizes volume increase 21.20% with being directly used in cotton than cotton stalk, and difference is extremely notable；Commercially available common organic fertilizer group
Ginnings is 3076.65kg/hm2, Trichoderma viride (Trichoderma viride) XJ-3 CGMCC are utilized with the present invention
NO.10989 prepares cotton stalk decomposed manure group significant difference, difference is organized compared with cotton stalk with being directly used in cotton not notable.To influenceing son
For cotton boll weight, the present invention prepares cotton stalk using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
Than cotton stalk, group adds 0.16g and 0.12g with being directly used in cotton respectively for decomposed manure group and commercially available common organic fertilizer group, not
Reach the significance level of difference；For single basal munure, the present invention utilizes Trichoderma viride (Trichoderma viride) XJ-3
CGMCC NO.10989 prepare cotton stalk decomposed manure group list basal munure 15.6, organized with being directly used in cotton compared with cotton stalk it is high by 2.40, it is poor
Heteropole is notable.From the data analysis of table 1, the present invention utilizes Trichoderma viride (Trichoderma viride) XJ-3 CGMCC
NO.10989, which prepares cotton stalk decomposed manure group, certain production-increasing function to cotton.
Influence of the different disposal of table 1 to output of cotton constituent element and yield
(4) present invention prepares cotton stalk using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
Influence of the decomposed manure group to Cotton Soil fertility
Table 2 shows that the present invention is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
Cotton stalk decomposed manure produces wholesome effect to soil fertility.The present invention utilizes Trichoderma viride (Trichoderma viride)
XJ-3 CGMCC NO.10989 prepare cotton stalk decomposed manure group quick-acting potassium content and organize raising with being directly used in cotton than cotton stalk
44.57%, and higher than commercially available common organic fertilizer group；The present invention utilizes Trichoderma viride (Trichoderma viride) XJ-3
CGMCC NO.10989 prepare cotton stalk decomposed manure group available phosphorus contents and organize raising 136.86% with being directly used in cotton compared with cotton stalk
And higher than commercially available common organic fertilizer group；Water-soluble calcium content organizes raising 215.38% with being directly used in cotton than cotton stalk, and is higher than city
Sell common organic fertilizer group；The present invention is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989
Cotton stalk decomposed manure can promote the organic matter decomposition in soil, and the content of organic matter is organized and reduced respectively with being directly used in cotton compared with cotton stalk
14.06%.
Influence of the different disposal of table 2 to Cotton Soil fertility
The Trichoderma viride of present invention screening seed selection domestication acquisition is obtained by above-mentioned series embodiment verification experimental verification
(Trichoderma viride) XJ-3 CGMCC NO.10989 bacterial strain has stronger growth and breeding ability, growth rate
It hurry up, hereditary capacity is stable, and to cotton growth and increases soil fertility with specific effect；By facts have proved profit of the invention
It is brown that cotton stalk decomposed manure group is prepared with Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989,
It is soft flexible when hand is grabbed, it applied to the soil on improvement cotton ground, can improve and activating soil, improve the fertility of soil, and
Cotton stalk decomposed manure group quick-acting potassium content prepared by the present invention organizes raising 44.57%, rapid available phosphorus with being directly used in cotton than cotton stalk
Content organizes raising 136.86% with being directly used in cotton compared with cotton stalk, and water-soluble calcium content organizes raising with being directly used in cotton than cotton stalk
215.38%, while the organic matter decomposition in soil can be promoted, the content of organic matter is organized and reduced respectively with being directly used in cotton compared with cotton stalk
14.06%；The cotton that the present invention is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989 bacterial strains
Bar decomposed manure can increase the yield of cotton, and seed cotton yield organizes volume increase 21.20%, unginned cotton's bell weight with being directly used in cotton than cotton stalk
Than cotton stalk, group adds 0.16g with being directly used in cotton, and single basal munure is 15.6, is organized with being directly used in cotton compared with cotton stalk high by 2.40；This
The cotton stalk that invention is prepared using Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989 bacterial strains is decomposed organic
The fertilizer group incidence of disease minimum 0.67%, the diseased plant rate of commercially available common organic fertilizer group is 1.33%, what cotton stalk was organized with being directly used in cotton
Diseased plant rate highest, it is 2.01%, the adverse effect during soil is produced to cotton planting reduces, to cotton in seedling stage damping-off, charcoal
Subcutaneous ulcer disease has certain prevention effect, therefore has extensively and applicable value in cotton planting and in increasing soil fertility.Meanwhile
The present invention uses the cotton stalk of Trichoderma viride (Trichoderma viride) XJ-3 CGMCC NO.10989 bacterial strains preparation is decomposed to have
Machine fertilizer, both solved the utilization of cotton stalk discarded object, reduced the pollution for burning cotton stalk to environment, while added the fertilizer in cotton soil
The quality of power, with can improveing cotton planting soil, is advantageous to the plantation of cotton, is advantageous to improve the yield and quality of cotton,
Good technique effect is obtained, meaning and the effect in improvement cotton planting ground with reality.
Above-described embodiment is only intended to clearly illustrate example of the present invention, and is not the restriction to embodiment.
For those of ordinary skill in the field, other various forms of changes can also be made on the basis of the above description
Or change.There is no necessity and possibility to exhaust all the enbodiments.And the obvious change thus extended
Or among changing still in protection scope of the present invention.
SEQUENCE LISTING
<110>Hami Xin Hemian industry limited company
<120>A kind of Trichoderma viride XJ-3 and its method for preparing cotton stalk decomposed manure
<130>Trichoderma viride
<160> 1
<170> PatentIn version 3.
<210> 1
<211> 597
<212> DNA
<213>Trichoderma viride
<220>
<221> ITS rDNA
<222> (1)..(597)
<400> 1
tccgtaggtg aacctgcgga gggatcatta ccgagtttac aactcccaaa ccccaatgtg 60
aacgttacca atctgttgcc tcggcgggat tctcttgccc cgggcgcgtc gcagccccgg 120
atcccatggc gcccgccgga ggaccaactc caaactcttt tttctctccg tcgcggctcc 180
cgtcgcggct ctgttttatt tttgctctga gcctttctcg gcgaccctag cgggcgtctc 240
gaaaatgaat caaaactttc aacaacggat ctcttggttc tggcatcgat gaagaacgca 300
gcgaaatgcg ataagtaatg tgaattgcag aattcagtga atcatcgaat ctttgaacgc 360
acattgcgcc cgccagtatt ctggcgggca tgcctgtccg agcgtcattt caaccctcga 420
acccctccgg ggggtcggcg ttggggatcg gcccctcacc gggccgcccc cgaaatacag 480
tggcggtctc gccgcagcct ctcctgcgca gtagtttgca cactcgcacc gggagcgcgg 540
cgcggccaca gccgtaaaac accccaaact tctgaaatgt gacctcggat caggtag 597
Claims (4)
1. a kind of Trichoderma viride (Trichoderma viride) XJ-3 for being applied to prepare cotton stalk decomposed manure, its feature exist
In described Trichoderma viride (Trichoderma viride) XJ-3 CGMCC culture presevation number is NO.10989.
2. Trichoderma viride (Trichoderma viride) XJ-3 as claimed in claim 1 is in cotton stalk decomposed manure is prepared
Using.
A kind of 3. method that cotton stalk decomposed manure is prepared using Trichoderma viride XJ-3, it is characterised in that described preparation method
Method acquisition is prepared by the following procedure：
(1) strain activation and culture：Prepare Trichoderma viride (Trichoderma viride) XJ-3CGMCC NO.10989 bacterial strains
Potato dextrose agar, sterile working is strictly carried out after sterilizing, flat board is forwarded to from inclined-plane, is cultivated at 28 DEG C of temperature
5 days；
(2) one-level culture：It is forwarded to and is equipped with from picking single bacterium colony on the potato dextrose agar solid medium in step (1)
In the 50mL conical flasks of potato dextrose broth, sterile working, 28 DEG C, 120r/min cultures 24h-48h；
(3) two level expands culture：Above-mentioned steps one-level culture strain is inoculated in and fills potato dextrose broth
In 500mL conical flasks, sterile working, 28 DEG C, 120r/min cultures 24h-48h；
(4) determination of water：Mark is assessed according to the ratio that rear every mu of cotton stalk yield of cotton is 250 kilograms to 450 kilograms is plucked
Standard, cotton stalk processing need to add the 60%-65% that water is cotton stalk yield；
(5) it is inoculated with：Trichoderma viride (Trichoderma viride) XJ-3CGMCC NO.10989 prepared by step (3) are activated
In water prepared by strain inoculation above-mentioned steps (4), water 10%-15% ratio inoculation as required prepares Trichoderma viride
Microbial inoculum, and strain protective agent, urea and diamines are added respectively according to the microbial inoculum amount of preparation, wherein, strain protective agent is by sucrose, paddy
Propylhomoserin sodium and glycerine are according to weight ratio 1:1:1 proportioning, strain protective agent is according to 15% addition, and urea and diamines are respectively according to 30%
Ratio with 20% is added, and fermentation base-material is prepared by stirring；
(6) cotton stalk crushing returning to the field：After having plucked cotton in autumn, by crop field operation by cotton stalk crushing, the length of cotton stalk after crushing
Control as 3cm-5cm, the fermentation base that step (5) preparation is sprayed on spray equipment cotton stalk after being pulverized is utilized in crushing process
Material；
(7) ferment：Fermentation base-material prepared by step (6) is raised together in company with the cotton stalk after crushing and is sprinkling upon in cotton field, is adopted
Buried, irrigate using winter irrigation water by cotton field flood is climing, autumn last year to Second Year with common equipment of the ploughing base-material that will ferment
Cotton stalk is prepared into cotton stalk decomposed manure by fermenting during spring.
4. the cotton stalk for preparing the method preparation of cotton stalk decomposed manure using Trichoderma viride XJ-3 as claimed in claim 3 is decomposed to be had
Machine fertilizer.
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| CN101492646A (en) * | 2009-01-15 | 2009-07-29 | 山东大学 | Trichoderma viride engineering bacterium and uses thereof |
| CN102071145A (en) * | 2010-09-09 | 2011-05-25 | 河南省农业科学院 | Trichoderma viride fungi and preparation and application of fungicide thereof |
| CN103397554A (en) * | 2013-07-29 | 2013-11-20 | 安徽安生生物化工科技有限责任公司 | New process for preparing microcrystalline cellulose from lignocelluloses biomass |
| CN104649757A (en) * | 2015-02-04 | 2015-05-27 | 中国烟草总公司广东省公司 | Soil high-carbon-based additive prepared from fermented cotton straw |
| CN104692846A (en) * | 2015-02-27 | 2015-06-10 | 山东省农业可持续发展研究所 | Straw bio-organic fertilizer and preparation method thereof |
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