CN105154336A - Trichoderma viride XJ-3 and method for preparing cotton stalk decomposed organic fertilizer by using same - Google Patents
Trichoderma viride XJ-3 and method for preparing cotton stalk decomposed organic fertilizer by using same Download PDFInfo
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- CN105154336A CN105154336A CN201510580276.6A CN201510580276A CN105154336A CN 105154336 A CN105154336 A CN 105154336A CN 201510580276 A CN201510580276 A CN 201510580276A CN 105154336 A CN105154336 A CN 105154336A
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Classifications
-
- 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
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses trichoderma viride XJ-3 and a method for preparing a cotton stalk decomposed organic fertilizer by using the same. The method for preparing the organic fertilizer with the decomposed cotton stalks has the outstanding technical effects of increasing the yield of cotton, improving the soil fertility and the like by separating, screening, breeding and domesticating strains of soil samples in the cotton field, determining Trichoderma viride (Trichoderma and viride) XJ-3CGMCC NO.10989 by rDNAITS sequence determination and phylogenetic analysis, inoculating the Trichoderma viride into crushed cotton stalks, wherein the crushed cotton stalks have the length of 3cm-5cm and the water content of 60-65 percent, adding 30 percent of urea, 20 percent of diammonium and a strain protective agent, preparing a bacterial solution, the urea, the diammonium and the strain protective agent into a fermentation base material, spraying the fermentation base material on the crushed stalks, turning the cotton field over for winter irrigation, fermenting for 3 months in autumn to the next spring.
Description
Technical field
The present invention relates to agriculture technical field of microbe application, specifically, the green wood enzyme and the viride XJ-3 that the present invention relates to a kind of seed selection voluntarily screening prepare the technical field of cotton stalk decomposed manure.
Background technology
Soil organic matter, as the important component in soil and Terrestrial Carbon storehouse, not only contributes to the Sustainable development of agricultural, and has impact to the capacity of carbon base in soil, release and turnover etc.Increase soil organic carbon, raising crop yield and Water and nitrogen use efficiency are the important goals of arid biogeographic zone farmland optimum management.Biological carbon is the organic carbon that a kind of carbon content is high, stability is strong, effectively can increase Soil carbon storage, increases soil fertility and productivity.
China is important cotton production areas, the world, and the cotton stalk after cotton harvesting is a kind of purposes precious resources widely.First, cotton stalk is a kind of well substitute for wood, and can be used as producing regenerated fiber, can manufacture the sheet materials such as shaving board, fiberboard and acoustic panel again simultaneously, or be used as building materials paper making raw material, quality outclass straw and straw; Secondly, cotton stalk has very high nutritive value, and after treatment with micron, its protein content can up to 12%, than wheat straw and straw protein content height 5-6 doubly, can be used as the first-class feed of cattle and sheep; Again, cotton stalk, through the chemical process such as grating and high temperature pyrolysis, produces the biogas based on carbon monoxide, can be used as a kind of novel energy; Cotton stalk also can be used as the substratum of pollution-free food edible mushrooms after pulverizing, the disposable tableware that this external application stalk is made is pollution-free, degradable and renewable.Therefore, from development green agriculture, cotton stalk can be agricultural deep processing after reclaiming provides sufficient with required starting material.
At present, the general treatment process of cotton stalk: is burn cotton stalk, and this is not only a kind of waste, and serious environment pollution; Two is the cotton stalks of hand harvest, and labour intensity is large, and efficiency is low; Three is direct chopping and returnings, though this way can increase the fertilizer of soil, can increase the sickness rate of cotton diseases and insect pests in the coming year, cotton stalk is not easily pulverized, and rely on natural condition merely, after pulverizing, cotton stalk corruption ability is weak, is difficult to be utilized by the cotton of next batch as fertilizer.Xinjiang, as the super-huge Cotton Production base of China, produces cotton stalk about 5.4 × 10 every year on average
6t.The biological carbon also field of cotton stalk pyrolysis being prepared, can reduce the disadvantageous effect of straw directly returning to field, effectively can improve soil productivity simultaneously.Although have many about utilizing cotton stalk to prepare substratum and studying compound manure for improving research and the report of cotton planting ground soil quality, but about separation screening fermentation capacity in the soil from cotton planting ground is strong and xylogen in cotton stalk of can degrading, the bacterial classification of the compositions such as Mierocrystalline cellulose, and utilize seed selection voluntarily screen domestication obtain viride carry out ferment cotton stalk prepare cotton stalk decomposed manure for improve cotton planting ground soil, increase the fertility of soil, to cotton in seedling stage damping-off, anthrax has certain preventive and therapeutic effect, the research being conducive to cotton growth have not been reported.
Summary of the invention
Also utilize the bacterial classification of screening to prepare the state of the art of cotton stalk decomposed manure about screening viride in the soil from cotton planting ground as excellent species for having no in prior art, the present invention aims to provide a kind of viride XJ-3 and prepares the method for cotton stalk decomposed manure.The present invention by isolating a collection of microorganism strains in the soil on cotton planting ground, therefrom separate the bacterial strain that a strain is numbered XJ-3, and by utilizing the viride being numbered XJ-3, add bacterial classification protective material, under the condition of urea and diamines, be that cotton stalk decomposed manure prepared by raw material with cotton stalk, reduce the disadvantageous effect of straw directly returning to field, effectively improve soil productivity, both the utilization of cotton stalk waste had been solved, reduce and burn cotton stalk to the pollution of environment, add the fertility in cotton soil simultaneously, the quality in soil, cotton planting ground can be improved, to cotton in seedling stage damping-off, anthrax has certain prevention effect, be conducive to the plantation of cotton, be conducive to the yield and quality improving cotton, and obtain good technique effect, improve the efficiency utilization rate of cotton stalk, there is the meaning and function of reality in improvement cotton planting ground.
The present invention adopts main technical scheme:
By the separation screening of microbial strains, cotton planting ground soil in isolate a collection of microorganism strains, through being separated further, screening, seed selection, domestication, obtain the trichoderma viride strain that a strain is numbered XJ-3.Measure and Phylogenetic Analysis by carrying out morphological specificity, physio-biochemical characteristics and rDNAITS sector sequence to obtained bacterial strain, tentatively determine its classification position.Simultaneously, the viride being numbered XJ-3 is inoculated in the cotton stalk after pulverizing, after wherein pulverizing, the length of cotton stalk is 3cm-5cm, water content is 60%-65%, and add the urea of 30%, two ammoniums of 20%, bacterial classification protective material, by bacterium liquid, urea, two ammoniums and bacterial classification protective material are prepared into fermentation base-material, and spray on stalk after being pulverized, plough after winter irrigation cottonly and ferment, the fermentative degradation time of 3 months is reached spring through autumn to Second Year, cotton stalk decomposed manure can be prepared, the cotton stalk decomposed manure of preparation is brown, when hand is grabbed, soft flexible, carry out fermentation from the bacterial classification of row filter and cotton stalk prepare cotton stalk decomposed manure by utilizing, both solved the utilization of cotton stalk waste, and reduced and burn cotton stalk to the pollution of environment, add the fertility in cotton soil simultaneously, the quality in soil, cotton planting ground can be improved, be conducive to the plantation of cotton, be conducive to the yield and quality improving cotton, obtain good technique effect, there is the meaning and function of reality in improvement cotton planting ground.
The present invention specifically provides a kind of viride (Trichodermaviride), by being separated the Soil Microorganism on cotton planting ground, screening and cultivating, obtain a collection of microorganism strains, therefrom filtering out a strain sequence number is the bacterial strain of XJ-3, through microbiological classification and qualification, this bacterial strain belongs to Trichodermaviride.
Concrete, the invention provides a kind of viride (Trichodermaviride), strain number is XJ-3.This bacterial strain was preserved in budapest treaty microorganism International Depository Authority before the applying date: China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC).Address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode: 100101.Preservation date is on June 23rd, 2015, and preserving number is CGMCCNo.10989.Trichodermaviride is accredited as through microbiology.This bacterial strain optimal culture conditions: temperature 25 DEG C-30 DEG C, pH is 5-5.5, substratum potato dextrose agar (PDA) substratum; Be white when this bacterial strain bacterium colony starts, fine and close, circular, to surrounding expansion, after produce green spores from bacterium colony central authorities, central authorities become green, the zone of growth of periphery of bacterial colonies adularescent mycelia, and last whole bacterium colony all becomes green; Spore is oval, CO
2cO is depended on the impact of this strain growth
2concentration and the pH value of substratum, in alkaline matrix, the CO of high density
2be conducive to the growth being numbered XJ-3 bacterial strain; This bacterial strain can utilize gas chromatography as carbon source, and comparatively ideal is monosaccharide and disaccharide, polysaccharide, purine, pyrimidine and amino acid etc.; This bacterial strain produces acids in a large number on the substratum being rich in carbohydrate, and with glucose or starch as carbon source, this bacterium produces the citric acid of 60%-80%; In buffer medium, ammonium is the nitrogenous source that this bacterial strain the most easily utilizes, and other nitrogenous sources such as amino acid, urea, nitrate, nitrite also can maintain its normal growth; Good especially with the growth of asparagus fern Men Suwei nitrogenous source, nitrogen content is low, can promote sporulation, to the negatively influencing of high concentration nitrate, because the existence of magnesium sulfate can be compensated; Inorganic salt and micro-inorganic salt very important to the growth of this bacterial strain, magnesium ion can promote that it grows, cupric ion can promote conidium chromogenesis, the formation of iron ion to spore has vital role, be accredited as Trichodermaviride bacterium through microbiology, but have obvious difference with common viride bacterial classification.With reference to " Fungal identification handbook ", the qualification of system Physiology and biochemistry is carried out to numbering XJ-3 bacterial strain, detect through Physiology and biochemistry and determine that numbering XJ-3 bacterial strain is the member in Trichoderma (Trichoderma).By the comparison of BLAST homology, after the ITSrDNA sequence nucleotide sequence of strain X J-3 carries out BLAST analysis in ncbi database, constructing system evolutionary tree, this is numbered strain X J-3 and TrichodermalongibrachiatumisolateT50 and is in minimum branch, be its allied species, its homology is 76%; And then this strain X J-3 is defined as Trichodermaviride, through said system taxonomy qualification classification, demonstrate viride provided by the invention (Trichodermaviride) and in physio-biochemical characteristics and molecular water equality, there is obvious otherness with common viride, according to bacterial classification Analysis of The Physiological And Biochemical Properties, from taxonomy, the comprehensive identification of molecular level analysis and systematics, identifies that this bacterial strain is viride (Trichodermaviride).
Meanwhile, the present invention specifically provides one to utilize viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare the method for cotton stalk decomposed manure, and concrete preparation method's step is as follows:
(1) strain activation and culture: the potato dextrose agar preparing viride (Trichodermaviride) XJ-3CGMCCNO.10989 bacterial strain, strictly aseptic technique is carried out after sterilizing, be forwarded to flat board from inclined-plane, cultivate 5 days at temperature 28 DEG C.
(2) one-level is cultivated: from the potato dextrose agar solid medium step (1), picking list bacterium colony is forwarded to and is equipped with in the 50mL Erlenmeyer flask of potato dextrose broth, aseptic technique, 28 DEG C, 120r/min cultivates 24h-48h.
(3) secondary enlarged culturing: above-mentioned steps one-level is cultivated strain inoculation in the 500mL Erlenmeyer flask filling potato dextrose broth, aseptic technique, 28 DEG C, 120r/min cultivates 24h-48h.
(4) determination of the water yield: be that the ratio of 250 kilograms to 450 kilograms assesses standard according to plucking rear every mu, cotton cotton stalk output, cotton stalk processing needs to add the 60%-65% that the water yield is cotton stalk output.
(5) inoculate: in water prepared by viride (Trichodermaviride) XJ-3CGMCCNO.10989 activated spawn inoculation above-mentioned steps (4) step (3) prepared, the ratio inoculation of water yield 10%-15% prepares viride microbial inoculum as required, and add bacterial classification protective material respectively according to the microbial inoculum amount of preparation, urea and diamines, wherein, bacterial classification protective material is by sucrose, Sodium Glutamate and glycerine are according to weight ratio 1:1:1 proportioning, bacterial classification protective material adds according to 15%, urea and diamines add according to the ratio of 30% and 20% respectively, fermentation base-material is prepared by stirring.
(6) cotton stalk crushing also field: after having plucked cotton in autumn, by land for growing field crops operation by cotton stalk crushing, after pulverizing, the cut to lengthen of cotton stalk is 3cm-5cm, utilizes fermentation base-material spray equipment cotton stalk after being pulverized spraying step (5) and prepare in crushing process.
(7) ferment: fermentation base-material step (6) prepared is raised together in company with the cotton stalk after pulverizing and is sprinkling upon in cotton field, common equipment of ploughing is adopted to be buried by fermentation base-material, utilize winter irrigation water to irrigate climing for cotton field flood, during autumn last year to Second Year spring, by fermentation, cotton stalk is prepared cotton stalk decomposed manure.
Further, the invention provides the cotton stalk decomposed manure adopting the above-mentioned method utilizing viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure to prepare.
The invention provides one utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare the method for cotton stalk decomposed manure, add urea and diamines, viride (Trichodermaviride) XJ-3CGMCCNO.10989 bacterial strain needs sufficient Carbon and nitrogen sources in its growth metabolism on the one hand, and interpolation urea and diamines are conducive to the Growth and reproduction of bacterial classification viride; Interpolation urea and diamines can promote the fermentation of cotton stalk on the other hand, also help the quality improving cotton planting ground and the growth promoting next batch of cotton simultaneously.
The invention provides one utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare the method for cotton stalk decomposed manure, add bacterial classification protective material, be conducive to the normal growth metabolism protecting viride (Trichodermaviride) XJ-3CGMCCNO.10989 bacterial strain on the one hand; On the other hand for the growth of bacterial strain provides nutritive substance, be conducive to the fermentation of cotton stalk.Add bacterial classification protective material by sucrose, Sodium Glutamate and glycerine according to weight ratio 1:1:1 proportioning; interpolation sucrose and Sodium Glutamate contribute to bacterial classification provides certain sugared source and energy element in cotton field in fermentation culture; help the breeding of bacterial classification; also bacterial classification can be made to obtain certain protection simultaneously; glycerine is adopted to guarantee bacterial classification large Tanaka along with a kind of temperature low sfgd. when the cotton stalk pulverized is survived the winter in cotton field; the fermentation of cotton stalk can be promoted, also help the quality improving cotton planting ground and the growth promoting next batch of cotton simultaneously.
In the present invention, potato glucose (PDA) Solid agar culture adopted, potato dextrose broth are all common substratum, its substratum content and component, and the culture condition of substratum, those of ordinary skill in the art can by looking in known technology reference book or document.
By implementing the concrete summary of the invention of the present invention, following beneficial effect can be reached:
(1) viride (Trichodermaviride) the XJ-3CGMCCNO.10989 bacterial strain of the present invention's screening has stronger growth and breeding ability, and growth velocity is fast, and hereditary property is stablized, and has specific effect to degraded cotton stalk.
(2) the present invention utilizes cotton stalk decomposed manure prepared by viride (Trichodermaviride) XJ-3CGMCCNO.10989 bacterial strain, in brown, when hand is grabbed, soft flexible.
(3) one of the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare the method for cotton stalk decomposed manure, both solved the utilization of cotton stalk waste, and reduced and burn cotton stalk to the pollution of environment, add the fertility in cotton soil simultaneously, the quality in soil, cotton planting ground can be improved, be conducive to the plantation of cotton, be conducive to the yield and quality improving cotton, obtain good technique effect, there is the meaning and function of reality in improvement cotton planting ground.
(4) one of the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare the method for cotton stalk decomposed manure, reduce the disadvantageous effect of straw directly returning to field, effectively improve soil productivity, improve the utilization ratio of cotton stalk simultaneously.
(5) method that one of the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure produces beneficial effect to soil fertility.Cotton stalk decomposed manure group quick-acting potassium content prepared by the present invention is directly used in than cotton stalk and cotton organizes raising 44.57%, available phosphorus contents comparatively cotton stalk is directly used in and organizes raising 136.86% cottonly, water-soluble calcium content is directly used in than cotton stalk and cotton organizes raising 215.38%, can impel the organic matter decomposition in soil, organic content comparatively cotton stalk is directly used in organize cottonly and reduces by 14.06% respectively simultaneously.
(6) method that one of the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure can increase the output of cotton, seed cotton yield is directly used in than cotton stalk and cotton organizes volume increase 21.20%, unginned cotton's bell anharmonic ratio cotton stalk is directly used in cotton ground group and adds 0.16g, single basal munure is 15.6, comparatively cotton stalk be directly used in organize cottonly high by 2.40, therefore, the cotton stalk decomposed manure group that prepared by the present invention has certain production-increasing function to cotton.
(7) method that one of the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure has certain prevention effect to cotton in seedling stage damping-off, anthrax.Wherein, cotton stalk decomposed manure group sickness rate prepared by the present invention is minimum is 0.67%, and the diseased plant rate of commercially available common fertilizer group is 1.33%, and it is the highest that cotton stalk is directly used in the diseased plant rate organized cottonly, is 2.01%.
Accompanying drawing explanation
The colonial morphology figure of the shown in green wood of Fig. 1 mould (Trichodermaviride) XJ-3CGMCCNO.10989, in figure, A is bacterium colony front, and B is the bacterium colony back side.
The spore micro-structure diagram of the shown in green wood of Fig. 2 mould (Trichodermaviride) XJ-3CGMCCNO.10989.
The mycelia micro-structure diagram of the shown in green wood of Fig. 3 mould (Trichodermaviride) XJ-3CGMCCNO.10989.
The rDNAITS sector sequence phylogeny tree graph of the shown in green wood of Fig. 4 mould (Trichodermaviride) XJ-3CGMCCNO.10989.
Embodiment
, for embodiment, the present invention is described below, but the present invention is not limited to following embodiment.
The all raw and auxiliary materials selected in the present invention, and the bacterium culture medium selected, cultural method are all well known selecting, the % related in the present invention is weight percentage, unless otherwise indicated.
Embodiment one: the separation of viride (Trichodermaviride) XJ-3CGMCCNO.10989, screening and qualification
1, the Isolation and screening of bacterial classification
(1) be separated
Viride used in the present invention carries out sampling and is separated from the soil of South Sinkiang, Xinjiang Cotton Cultivation in Akesu Region planting site, and according to strain degradation Analysis on action mechanism, preliminary screening goes out bacterial strain cotton stalk to certain degradation capability.Traditional plating method is utilized to isolate microorganism in soil layer, plate streak purifying bacterial strain, with different culture temperature, pH value, substratum for enrichment condition, filter out a collection of well-grown microorganism strains, therefrom optimizing a strain sequence number is the bacterial strain of XJ-3.
Separating step: according to gradient dilution method, takes pedotheque in 10g cotton field and, in 90mL stroke-physiological saline solution, carries out gradient dilution, choose 10 after 30 DEG C of activation 30min
-2, 10
-3, 10
-4diluent coats the flat board of potato dextrose agar (PDA) substratum respectively, each process 3 repetition, in 28 DEG C of cultivations.The bacterium colony that picking shape, size, color etc. are different after growing bacterium colony distinguishes streak inoculation in new isolation medium potato dextrose agar (PDA) substratum, until fall without miscellaneous bacteria.A bacterial strain part after purifying is adopted the mode preservations such as lyophilized products ampoul tube, glycerine pipe and liquid nitrogen, and a part is stored in 4 DEG C and is directly used in follow-up study.
(2) culture condition
By the inoculation of purifying in potato dextrose agar (PDA) culture medium slant, cultivate 48h in 28 DEG C, put into 4 DEG C of refrigerators and save backup.
Growth temperature: the growth temperature range being numbered XJ-3 bacterial strain is 20 DEG C-38 DEG C, and optimum temperuture is 25 DEG C-30 DEG C.
Growth pH: the growth pH value being numbered XJ-3 bacterial strain be 1.5 or 9.0 substratum on can grow, but acidic conditions is higher than the germination rate under alkaline condition, and growth pH scope is 1.5-9.0, and the most suitable growth pH value is 5-5.5.
Carbon source: be numbered XJ-3 bacterial strain and gas chromatography can be utilized as carbon source, comparatively ideal is monosaccharide and disaccharide, polysaccharide, purine, pyrimidine and amino acid etc.
Nitrogenous source: in buffer medium, ammonium is numbered the nitrogenous source that XJ-3 bacterial strain the most easily utilizes, and other nitrogenous source such as amino acid, urea, nitrate, nitrite all can maintain its normal growth; Time wherein with asparagus fern Men Suwei nitrogenous source, strain growth is good, and nitrogen content is low, can promote sporulation, and magnesium sulfate can reduce high concentration nitrate to the negatively influencing being numbered XJ-3 bacterial strain.
Inorganic salt and trace element: magnesium ion can promote to be numbered XJ-3 strain growth, cupric ion can promote to be numbered XJ-3 bacterial strain conidium chromogenesis, and the formation of iron ion to spore has vital role.
Being prepared as of described potato dextrose agar (PDA) substratum: potato is cleaned peeling, gets 200 grams and be cut into small pieces, add water 1000 milliliters, after boiling half an hour, supply moisture.In filtrate, add 10 grams of agar, to boil after dissolving with sucrose 20 grams, supply moisture, packing, sterilizing, PDA solid medium can be prepared.
By being separated the Soil Microorganism on cotton planting ground, screening and cultivating, obtain a collection of microorganism strains, therefrom filtering out a strain sequence number is the bacterial strain of XJ-3, and through microbiological classification and qualification, this bacterial strain belongs to Trichodermaviride.The invention provides a kind of viride (Trichodermaviride), strain number is XJ-3.This bacterial strain was preserved in budapest treaty microorganism International Depository Authority before the applying date: China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC).Address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode: 100101.Preservation date is on June 23rd, 2015, and preserving number is CGMCCNo.10989.Trichodermaviride is accredited as through microbiology.This bacterial strain optimal culture conditions: temperature 25 DEG C-30 DEG C, pH is 5-5.5, and substratum is potato dextrose agar (PDA) substratum; Be white when this bacterial strain bacterium colony starts, fine and close, circular, to surrounding expansion, after produce green spores from bacterium colony central authorities, central authorities become green, the zone of growth of periphery of bacterial colonies adularescent mycelia, and last whole bacterium colony all becomes green; Spore is oval, CO
2cO is depended on the impact of this strain growth
2concentration and the pH value of substratum, in alkaline matrix, the CO of high density
2be conducive to the growth being numbered XJ-3 bacterial strain; This bacterial strain can utilize gas chromatography as carbon source, and comparatively ideal is monosaccharide and disaccharide, polysaccharide, purine, pyrimidine and amino acid etc.; This bacterial strain produces a large amount of acid on the substratum being rich in carbohydrate, and with glucose or starch as carbon source, this bacterium produces the citric acid of 60%-80%; In buffer medium, ammonium is the nitrogenous source that this bacterial strain the most easily utilizes, and other nitrogenous sources such as amino acid, urea, nitrate, nitrite also can maintain its normal growth; During with asparagus fern Men Suwei nitrogenous source, it is good especially to grow, and nitrogen content is low, can promote sporulation; The existence of magnesium sulfate can reduce the negatively influencing of high concentration nitrate to this bacterial strain, owing to being compensated; Inorganic salt and micro-inorganic salt very important to the growth of this bacterial strain, magnesium ion can promote that it grows, cupric ion can promote conidium chromogenesis, the formation of iron ion to spore has vital role, be accredited as Trichodermaviride bacterium through microbiology, but have the difference of obvious colonial morphology, Physiology and biochemistry with common viride bacterial classification.With reference to " Fungal identification handbook ", the qualification of system Physiology and biochemistry is carried out to numbering XJ-3 bacterial strain, detect through Physiology and biochemistry and determine that numbering XJ-3 bacterial strain is the member in Trichoderma (Trichoderma).By the comparison of BLAST homology, after the ITSrDNA sequence nucleotide sequence of strain X J-3 carries out BLAST analysis in ncbi database, constructing system evolutionary tree, this is numbered strain X J-3 and TrichodermalongibrachiatumisolateT50 and is in minimum branch, be its allied species, its homology is 76%, and then this strain X J-3 is defined as Trichodermaviride, through said system taxonomy qualification classification, demonstrate the otherness that viride provided by the invention (Trichodermaviride) and common viride have obvious physio-biochemical characteristics difference and molecular level, according to bacterial classification Analysis of The Physiological And Biochemical Properties, the comprehensive identification of molecular level analysis and systematics, although the bacterial classification being numbered XJ-3 has obvious difference with common viride in physio-biochemical characteristics and molecular level, it is a kind of new bacterium, but from taxonomy, be accredited as viride (Trichodermaviride), see accompanying drawing 1, 2 and accompanying drawing 3.
Embodiment two: the separation of viride (Trichodermaviride) XJ-3CGMCCNO.10989, screening and qualification
1, the rDNAITS section of pcr amplification viride and order-checking thereof
Single bacterium colony of picking a small amount of XJ-3 bacterial strain, put into the EP pipe filling 25 μ L sterilized waters, 100 DEG C are boiled 8-10min, put into rapidly mixture of ice and water 5min afterwards.Centrifugal 10000r/min, 5min, 4 DEG C of preservations, the used time gets supernatant.
The structure of rDNAITS section gene sequencing and systematic evolution tree thereof: the DNA extracting bacterial strain according to a conventional method, will dilute universal primer with deionized water, and performing PCR of going forward side by side increases, and design of primers is as follows:
ITS1(F)：5'-TCCGTAGGTGAACCTGCGG-3'
ITS4(R)：5'-TCCTCCGCTTATTGATATGC-3'
The structure of rDNAITS section gene sequencing and systematic evolution tree thereof: the STb gene extracting bacterial strain according to a conventional method, with the pcr amplification of deionized water by dilution universal primer rDNAITS section.Primer through synthesis, electrophoresis detection, and through order-checking.
2, the comparison of rDNAITS section gene order and Phylogenetic Analysis
Nucleotide sequence in the rDNAITS section gene order and GenBank database that obtain checking order carries out BLAST analysis, therefrom obtain close rDNAITS section gene order, after the rDNAITS section gene order of XJ-3 is carried out BLAST analysis in ncbi database, constructing system evolutionary tree.Shown in accompanying drawing 4, between strain X J-3 and TrichodermalongibrachiatumisolateT50, evolutionary distance is the shortest, is the allied species of Trichodermalongibrachiatumisolate.In conjunction with Morphologic Characteristics and the physio-biochemical characteristics of XJ-3, determine that it is viride (Trichodermaviride) and belong to.By measured rDNAITS section gene order input Genbank, tetraploid rice is carried out with Blast program, find that the similarity of the rDNAITS section gene order of it and TrichodermalongibrachiatumisolateT50 is maximum, similarity is 76%, thus be defined as Trichodermaviride further, in physio-biochemical characteristics difference and molecular water equality, having obvious otherness with common viride, is a kind of typical new bacterium.In conjunction with Morphologic Characteristics and the physio-biochemical characteristics of XJ-3, from classification evolution angle, to determine that it is bacterium numbering be XJ-3 comprehensive identification is viride (Trichodermaviride)
Embodiment three: utilize viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure
Viride (Trichodermaviride) XJ-3CGMCCNO.10989 is utilized to prepare concrete preparation method's step of cotton stalk decomposed manure as follows:
(1) strain activation and culture: the potato dextrose agar preparing viride (Trichodermaviride) XJ-3CGMCCNO.10989 bacterial strain, strictly aseptic technique is carried out after sterilizing, be forwarded to flat board from inclined-plane, cultivate 5 days at temperature 28 DEG C.
(2) one-level is cultivated: from the potato dextrose agar solid medium step (1), picking list bacterium colony is forwarded to and is equipped with in the 50mL Erlenmeyer flask of potato dextrose broth, aseptic technique, 28 DEG C, 120r/min cultivates 24h-48h.
(3) secondary enlarged culturing: above-mentioned steps one-level is cultivated strain inoculation in the 500mL Erlenmeyer flask filling potato dextrose broth, aseptic technique, 28 DEG C, 120r/min cultivates 24h-48h.
(4) determination of the water yield: be that the ratio of 250 kilograms to 450 kilograms assesses standard according to plucking rear every mu, cotton cotton stalk output, cotton stalk processing needs to add the 60%-65% that the water yield is cotton stalk output.
(5) inoculate: in water prepared by viride (Trichodermaviride) XJ-3CGMCCNO.10989 activated spawn inoculation above-mentioned steps (4) step (3) prepared, the ratio inoculation of water yield 10%-15% prepares viride microbial inoculum as required, and add bacterial classification protective material respectively according to the microbial inoculum amount of preparation, urea and diamines, wherein, bacterial classification protective material is by sucrose, Sodium Glutamate and glycerine are according to weight ratio 1:1:1 proportioning, bacterial classification protective material adds according to 15%, urea and diamines add according to the ratio of 30% and 20% respectively, fermentation base-material is prepared by stirring.
(6) cotton stalk crushing also field: after having plucked cotton in autumn, by land for growing field crops operation by cotton stalk crushing, after pulverizing, the cut to lengthen of cotton stalk is 3cm-5cm, utilizes fermentation base-material spray equipment cotton stalk after being pulverized spraying step (5) and prepare in crushing process.
(7) ferment: fermentation base-material step (6) prepared is raised together in company with the cotton stalk after pulverizing and is sprinkling upon in cotton field, common equipment of ploughing is adopted to be buried by fermentation base-material, utilize winter irrigation water to irrigate climing for cotton field flood, during autumn last year to Second Year spring, by fermentation, cotton stalk is prepared cotton stalk decomposed manure.
In the present invention, the cotton stalk decomposed manure adopting viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare presents brown, when hand is grabbed, soft flexible.The present invention carries out fermentation from the bacterial classification of row filter and cotton stalk prepare cotton stalk decomposed manure by utilizing, and has both solved the utilization of cotton stalk waste, has reduced and burn cotton stalk to the pollution of environment, added the fertility in cotton soil simultaneously, be conducive to the plantation of cotton.
Embodiment four: utilize viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure
Viride (Trichodermaviride) XJ-3CGMCCNO.10989 is utilized to prepare concrete preparation method's step of cotton stalk decomposed manure as follows:
(1) strain activation and culture: the potato dextrose agar preparing viride (Trichodermaviride) XJ-3CGMCCNO.10989 bacterial strain, strictly aseptic technique is carried out after sterilizing, be forwarded to flat board from inclined-plane, cultivate 5 days at temperature 28 DEG C.
(2) one-level is cultivated: from the potato dextrose agar solid medium step (1), picking list bacterium colony is forwarded to and is equipped with in the 50mL Erlenmeyer flask of potato dextrose broth, aseptic technique, 28 DEG C, 120r/min cultivates 24h-48h.
(3) secondary enlarged culturing: above-mentioned steps one-level is cultivated strain inoculation in the 500mL Erlenmeyer flask filling potato dextrose broth, aseptic technique, 28 DEG C, 120r/min cultivates 24h-48h.
(4) determination of the water yield: be that the ratio of 250 kilograms assesses standard according to plucking rear every mu, cotton cotton stalk output, it is 60% of cotton stalk output that cotton stalk processing needs to add the water yield, and therefore the water yield is 150L.
(5) inoculate: in water prepared by viride (Trichodermaviride) XJ-3CGMCCNO.10989 activated spawn inoculation above-mentioned steps (4) step (3) prepared, the ratio inoculation of the water yield 10% prepares viride microbial inoculum as required, the bacterium liquid of access is 15L, and add bacterial classification protective material respectively according to the microbial inoculum amount of preparation, urea and diamines, wherein, bacterial classification protective material is by sucrose, Sodium Glutamate and glycerine are according to weight ratio 1:1:1 proportioning, bacterial classification protective material adds according to 15%, addition is 24.75 kilograms, urea and diamines add according to the ratio of 30% and 20% respectively, addition is 49.5 kilograms and 53 kilograms, and stir and prepare fermentation base-material.
(6) cotton stalk crushing also field: after having plucked cotton in autumn, by land for growing field crops operation by cotton stalk crushing, after pulverizing, the cut to lengthen of cotton stalk is 3cm-5cm, utilizes the fermentation base-material that spray equipment will the cotton stalk after pulverizing spray step (5) and prepares in crushing process.
(7) ferment: fermentation base-material step (6) prepared is raised together in company with the cotton stalk after pulverizing and is sprinkling upon in cotton field, common equipment of ploughing is adopted to be buried by fermentation base-material, utilize winter irrigation water to irrigate climing for cotton field flood, during autumn last year to Second Year spring, by fermentation, cotton stalk is prepared cotton stalk decomposed manure.
The present invention, when fermentation is for cotton stalk decomposed manure, adds bacterial classification protective material, is conducive to the normal growth metabolism protecting viride (Trichodermaviride) XJ-3CGMCCNO.10989 bacterial strain on the one hand; On the other hand for the growth of bacterial strain provides nutritive substance, be conducive to the fermentation of cotton stalk.Add bacterial classification protective material by sucrose, Sodium Glutamate and glycerine according to weight ratio 1:1:1 proportioning; interpolation sucrose and Sodium Glutamate contribute to bacterial classification provides certain sugared source and energy element in cotton field in fermentation culture; help the breeding of bacterial classification; also bacterial classification can be made to obtain certain protection simultaneously; glycerine is adopted to guarantee bacterial classification large Tanaka along with a kind of temperature low sfgd. when the cotton stalk pulverized is survived the winter in cotton field; the fermentation of cotton stalk can be promoted, also help the quality improving cotton planting ground and the growth promoting next batch of cotton simultaneously.
Embodiment five: utilize viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure
Viride (Trichodermaviride) XJ-3CGMCCNO.10989 is utilized to prepare concrete preparation method's step of cotton stalk decomposed manure as follows:
(1) strain activation and culture: the potato dextrose agar preparing viride (Trichodermaviride) XJ-3CGMCCNO.10989 bacterial strain, strictly aseptic technique is carried out after sterilizing, be forwarded to flat board from inclined-plane, cultivate 5 days at temperature 28 DEG C.
(2) one-level is cultivated: from the potato dextrose agar solid medium step (1), picking list bacterium colony is forwarded to and is equipped with in the 50mL Erlenmeyer flask of potato dextrose broth, aseptic technique, 28 DEG C, 120r/min cultivates 24h-48h.
(3) secondary enlarged culturing: above-mentioned steps one-level is cultivated strain inoculation in the 500mL Erlenmeyer flask filling potato dextrose broth, aseptic technique, 28 DEG C, 120r/min cultivates 24h-48h.
(4) determination of the water yield: be that the ratio of 450 kilograms assesses standard according to plucking rear every mu, cotton cotton stalk output, it is 65% of cotton stalk output that cotton stalk processing needs to add the water yield, and therefore the water yield is 292.5L.
(5) inoculate: in water prepared by viride (Trichodermaviride) XJ-3CGMCCNO.10989 activated spawn inoculation above-mentioned steps (4) step (3) prepared, the ratio inoculation of the water yield 15% prepares viride microbial inoculum as required, the bacterium liquid of access is 43.88L, and add bacterial classification protective material respectively according to the microbial inoculum amount of preparation, urea and diamines, wherein, bacterial classification protective material is by sucrose, Sodium Glutamate and glycerine are according to weight ratio 1:1:1 proportioning, bacterial classification protective material adds according to 15%, addition is 50.46 kilograms, urea and diamines add according to the ratio of 30% and 20% respectively, addition is 100.91 kilograms and 67.28 kilograms, and stir and prepare fermentation base-material.
(6) cotton stalk crushing also field: after having plucked cotton in autumn, by land for growing field crops operation by cotton stalk crushing, after pulverizing, the cut to lengthen of cotton stalk is 3cm-5cm, utilizes the fermentation base-material that spray equipment will the cotton stalk after pulverizing spray step (5) and prepares in crushing process.
(7) ferment: fermentation base-material step (6) prepared is raised together in company with the cotton stalk after pulverizing and is sprinkling upon in cotton field, common equipment of ploughing is adopted to be buried by fermentation base-material, utilize winter irrigation water to irrigate climing for cotton field flood, during autumn last year to Second Year spring, by fermentation, cotton stalk is prepared cotton stalk decomposed manure.
Embodiment six: utilize viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure
Viride (Trichodermaviride) XJ-3CGMCCNO.10989 is utilized to prepare concrete preparation method's step of cotton stalk decomposed manure as follows:
(1) strain activation and culture: the potato dextrose agar preparing viride (Trichodermaviride) XJ-3CGMCCNO.10989 bacterial strain, strictly aseptic technique is carried out after sterilizing, be forwarded to flat board from inclined-plane, cultivate 5 days at temperature 28 DEG C.
(2) one-level is cultivated: from the potato dextrose agar solid medium step (1), picking list bacterium colony is forwarded to and is equipped with in the 50mL Erlenmeyer flask of potato dextrose broth, aseptic technique, 28 DEG C, 120r/min cultivates 24h-48h.
(3) secondary enlarged culturing: above-mentioned steps one-level is cultivated strain inoculation in the 500mL Erlenmeyer flask filling potato dextrose broth, aseptic technique, 28 DEG C, 120r/min cultivates 24h-48h.
(4) determination of the water yield: be that the ratio of 350 kilograms assesses standard according to plucking rear every mu, cotton cotton stalk output, it is 62% of cotton stalk output that cotton stalk processing needs to add the water yield, and therefore the water yield is 217L.
(5) inoculate: in water prepared by viride (Trichodermaviride) XJ-3CGMCCNO.10989 activated spawn inoculation above-mentioned steps (4) step (3) prepared, the ratio inoculation of the water yield 12% prepares viride microbial inoculum as required, the bacterium liquid of access is 26.04L, and add bacterial classification protective material respectively according to the microbial inoculum amount of preparation, urea and diamines, wherein, bacterial classification protective material is by sucrose, Sodium Glutamate and glycerine are according to weight ratio 1:1:1 proportioning, bacterial classification protective material adds according to 15%, addition is 36.46 kilograms, urea and diamines add according to the ratio of 30% and 20% respectively, addition is 72.91 kilograms and 48.61 kilograms, and stir and prepare fermentation base-material.
(6) cotton stalk crushing also field: after having plucked cotton in autumn, by land for growing field crops operation by cotton stalk crushing, after pulverizing, the cut to lengthen of cotton stalk is 3cm-5cm, utilizes the fermentation base-material that spray equipment will the cotton stalk after pulverizing spray step (5) and prepares in crushing process.
(7) ferment: fermentation base-material step (6) prepared is raised together in company with the cotton stalk after pulverizing and is sprinkling upon in cotton field, common equipment of ploughing is adopted to be buried by fermentation base-material, utilize winter irrigation water to irrigate climing for cotton field flood, during autumn last year to Second Year spring, by fermentation, cotton stalk is prepared cotton stalk decomposed manure.
Embodiment seven: cotton stalk decomposed manure is conducive to the field experiment of cotton growth
1, test design and process
This test is carried out in the cotton planting ground of Aksu Prefecture.Before this research, planted continuously cottonly 2 season cotton, wherein earth type ploughs desert grey soil for filling with, and quality is attached most importance to earth, organic 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.
Test have 2 process and 1 contrast, that is: cotton stalk be directly used in cotton ground group, commercially available common fertilizer group, the present invention utilize viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure group.Cotton planting adopts plastic-film-covered cultivation, and film 4 row, Spacing form is 30cm+60cm+30cm, spacing in the rows 10cm, thickness of sowing 22.2 × 10
4strain/hm
2.Irrigation method is under-film drip irrigation, and a film two is managed, drip irrigation zone spacing 90cm.Adopt dry broadcasting to wet out, after planting ooze Miao Shui 45mm.Pour water during cotton growth 9 times, total irrigation volume 600mm, other cultivation management measure is with reference to local land for growing field crops.
2, process of the test
At the Different growth phases locality upper plant sample of cotton, be separated according to stem, leaf, cotton buds and bolls Different Organs, after the 30min that completes at 105 DEG C, then dry to constant weight at 70 DEG C, finally weigh.
At cotton in seedling stage, cotton disease is investigated; Divide community to receive flower meter to produce, mensuration bell weight; The fertility of soil.
3, test-results
(1) dry-matter accumulation change
Totally it seems, cotton dry matter content extends with cotton growing stage, and cotton stalk is directly used in the difference that cotton ground group, commercially available common fertilizer group, the present invention utilize viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare between cotton stalk decomposed manure group and constantly increases.Cotton stalk is directly used in be organized cotton dry-matter accumulation within vegetative period cottonly and increases mild; But commercially available common fertilizer group and the present invention utilize viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure group dry-matter accumulation after cotton flowering period and increase sharply, gradually mild after Shengjing Town.Meanwhile, the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare the dry matter accumulation amount of dry matter accumulation amount higher than commercially available common fertilizer group cotton of cotton stalk decomposed manure group cotton.
(2) the shadow noon of disease in cotton seedling stage
Sprout term disease is anthrax and damping-off mainly.The sickness rate that the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure group is all directly used in cotton group and commercially available common fertilizer group lower than cotton stalk, wherein the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure group sickness rate minimum is 0.67%, the diseased plant rate of commercially available common fertilizer group is 1.33%, it is the highest that cotton stalk is directly used in the diseased plant rate organized cottonly, is 2.01%.Result shows that the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure group and has certain prevention effect to cotton in seedling stage damping-off, anthrax.
(3) the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare the impact of cotton stalk decomposed manure on output of cotton constituent element
The present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure group seed cotton yield and is directly used in than cotton stalk and cotton organizes volume increase 21.20%, and difference is extremely remarkable; Commercially available common fertilizer group seed cotton yield is 3076.65kg/hm
2, utilize viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure group significant difference with the present invention, comparatively cotton stalk is directly used in that to organize difference not remarkable cottonly.Heavy for affecting unginned cotton's bell, the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure group and commercially available common fertilizer group and is directly used in cotton ground group than cotton stalk respectively and adds 0.16g and 0.12g, does not all reach the significance level of difference; With regard to single basal munure, the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure group list basal munure 15.6, and comparatively cotton stalk is directly used in and organizes high by 2.40 cottonly, and difference is extremely remarkable.From table 1 data analysis, the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure group has certain production-increasing function to cotton.
Table 1 different treatment is on the impact of output of cotton constituent element and output
(4) the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure group to the impact of Cotton Soil fertility
Table 2 shows that the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure and produces wholesome effect to soil fertility.The present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure group quick-acting potassium content and is directly used in than cotton stalk and cotton organizes raising 44.57%, and higher than commercially available common fertilizer group; The present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure group available phosphorus contents and is directly used in compared with cotton stalk and cotton organizes raising 136.86% and higher than commercially available common fertilizer group; Water-soluble calcium content is directly used in than cotton stalk and cotton organizes raising 215.38%, and higher than commercially available common fertilizer group; The present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure can impel organic matter decomposition in soil, and organic content comparatively cotton stalk is directly used in organize cottonly and reduces by 14.06% respectively.
Table 2 different treatment is on the impact of Cotton Soil fertility
Obtain the present invention by above-mentioned series embodiment verification experimental verification to screen the bacterial strain of viride (Trichodermaviride) XJ-3CGMCCNO.10989 that seed selection domestication obtains there is stronger growth and breeding ability, growth velocity is fast, hereditary property is stablized, and has specific effect to cotton growth with increasing soil fertility, be brown by facts have proved that the present invention utilizes viride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed manure group, when hand is grabbed, soft flexible, be applied to the soil on the cotton ground of improvement, 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 is directly used in than cotton stalk and cotton organizes raising 44.57%, available phosphorus contents comparatively cotton stalk is directly used in and organizes raising 136.86% cottonly, water-soluble calcium content is directly used in than cotton stalk and cotton organizes raising 215.38%, the organic matter decomposition in soil can be impelled simultaneously, organic content comparatively cotton stalk is directly used in organize cottonly and reduces by 14.06% respectively, the cotton stalk decomposed manure that the present invention adopts viride (Trichodermaviride) XJ-3CGMCCNO.10989 bacterial strain to prepare can increase the output of cotton, seed cotton yield is directly used in than cotton stalk and cotton organizes volume increase 21.20%, unginned cotton's bell anharmonic ratio cotton stalk is directly used in cotton ground group and adds 0.16g, single basal munure is 15.6, comparatively cotton stalk be directly used in organize cottonly high by 2.40, the cotton stalk decomposed manure group sickness rate that the present invention adopts viride (Trichodermaviride) XJ-3CGMCCNO.10989 bacterial strain to prepare is minimum is 0.67%, the diseased plant rate of commercially available common fertilizer group is 1.33%, it is the highest that cotton stalk is directly used in the diseased plant rate organized cottonly, be 2.01%, disadvantageous effect in being produced cotton planting by soil reduces, there is certain prevention effect to cotton in seedling stage damping-off, anthrax, therefore at cotton planting with in increasing soil fertility, there is extensive and applicable value.Simultaneously, the cotton stalk decomposed manure that the present invention adopts viride (Trichodermaviride) XJ-3CGMCCNO.10989 bacterial strain to prepare, both solved the utilization of cotton stalk waste, and reduced and burn cotton stalk to the pollution of environment, add the fertility in cotton soil simultaneously, the quality in soil, cotton planting ground can be improved, be conducive to the plantation of cotton, be conducive to the yield and quality improving cotton, obtain good technique effect, there is the meaning and function of reality in improvement cotton planting ground.
Above-described embodiment is only for example of the present invention is clearly described, and the restriction not to embodiment.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here exhaustive without the need to also giving all embodiments.And apparent change extended thus or variation are still among protection scope of the present invention.
SEQUENCELISTING
<110> Hami Xin Hemian industry limited-liability company
<120> viride XJ-3 and prepare the method for cotton stalk decomposed manure
<130> viride
<160>1
<170>PatentInversion3.
<210>1
<211>597
<212>DNA
<213> viride
<220>
<221>ITSrDNA
<222>(1)..(597)
<400>1
tccgtaggtgaacctgcggagggatcattaccgagtttacaactcccaaaccccaatgtg60
aacgttaccaatctgttgcctcggcgggattctcttgccccgggcgcgtcgcagccccgg120
atcccatggcgcccgccggaggaccaactccaaactcttttttctctccgtcgcggctcc180
cgtcgcggctctgttttatttttgctctgagcctttctcggcgaccctagcgggcgtctc240
gaaaatgaatcaaaactttcaacaacggatctcttggttctggcatcgatgaagaacgca300
gcgaaatgcgataagtaatgtgaattgcagaattcagtgaatcatcgaatctttgaacgc360
acattgcgcccgccagtattctggcgggcatgcctgtccgagcgtcatttcaaccctcga420
acccctccggggggtcggcgttggggatcggcccctcaccgggccgcccccgaaatacag480
tggcggtctcgccgcagcctctcctgcgcagtagtttgcacactcgcaccgggagcgcgg540
cgcggccacagccgtaaaacaccccaaacttctgaaatgtgacctcggatcaggtag597
Claims (5)
1. the viride being applicable to prepare cotton stalk decomposed manure (
trichodermaviride) XJ-3, it is characterized in that, described viride (
trichodermaviride) the CGMCC culture presevation number of XJ-3 is NO.10989.
2. as claimed in claim 1 viride (
trichodermaviride) gene order of XJ-3 is as SEQUENCELISTING.
3. as claimed in claim 1 viride (
trichodermaviride) XJ-3 preparing the application in cotton stalk decomposed manure.
4. utilize viride XJ-3 to prepare a method for cotton stalk decomposed manure, it is characterized in that, described preparation method is obtained by following preparation method:
(1) strain activation and culture: prepare viride (
trichodermaviride) potato dextrose agar of XJ-3CGMCCNO.10989 bacterial strain, strictly carry out aseptic technique after sterilizing, be forwarded to flat board from inclined-plane, cultivate 5 days at temperature 28 DEG C;
(2) one-level is cultivated: from the potato dextrose agar solid medium step (1), picking list bacterium colony is forwarded to and is equipped with in the 50mL Erlenmeyer flask of potato dextrose broth, aseptic technique, 28 DEG C, 120r/min cultivates 24h-48h;
(3) secondary enlarged culturing: above-mentioned steps one-level is cultivated strain inoculation in the 500mL Erlenmeyer flask filling potato dextrose broth, aseptic technique, 28 DEG C, 120r/min cultivates 24h-48h;
(4) determination of the water yield: be that the ratio of 250 kilograms to 450 kilograms assesses standard according to plucking rear every mu, cotton cotton stalk output, cotton stalk processing needs to add the 60%-65% that the water yield is cotton stalk output;
(5) inoculate: viride prepared by step (3) (
trichodermaviride) in XJ-3CGMCCNO.10989 activated spawn inoculation above-mentioned steps (4) water prepared, the ratio inoculation of water yield 10%-15% prepares viride microbial inoculum as required, and add bacterial classification protective material, urea and diamines respectively according to the microbial inoculum amount of preparation, wherein, bacterial classification protective material by sucrose, Sodium Glutamate and glycerine according to weight ratio 1:1:1 proportioning, bacterial classification protective material adds according to 15%, urea and diamines add according to the ratio of 30% and 20% respectively, prepare fermentation base-material by stirring;
(6) cotton stalk crushing also field: after having plucked cotton in autumn, by land for growing field crops operation by cotton stalk crushing, after pulverizing, the cut to lengthen of cotton stalk is 3cm-5cm, utilizes fermentation base-material spray equipment cotton stalk after being pulverized spraying step (5) and prepare in crushing process;
(7) ferment: fermentation base-material step (6) prepared is raised together in company with the cotton stalk after pulverizing and is sprinkling upon in cotton field, common equipment of ploughing is adopted to be buried by fermentation base-material, utilize winter irrigation water to irrigate climing for cotton field flood, during autumn last year to Second Year spring, by fermentation, cotton stalk is prepared cotton stalk decomposed manure.
5. cotton stalk decomposed manure prepared by the method utilizing viride XJ-3 to prepare cotton stalk decomposed manure as claimed in claim 4.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112852666A (en) * | 2021-01-19 | 2021-05-28 | 新疆河润水业有限责任公司 | Preparation method of microbial agent and microbial fertilizer prepared by adopting microbial agent |
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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 |
-
2015
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112852666A (en) * | 2021-01-19 | 2021-05-28 | 新疆河润水业有限责任公司 | Preparation method of microbial agent and microbial fertilizer prepared by adopting microbial agent |
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