CN108085304A - Soil-repairing agent and its production method and application of the height containing polyphenol oxidase - Google Patents
Soil-repairing agent and its production method and application of the height containing polyphenol oxidase Download PDFInfo
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- CN108085304A CN108085304A CN201711459192.2A CN201711459192A CN108085304A CN 108085304 A CN108085304 A CN 108085304A CN 201711459192 A CN201711459192 A CN 201711459192A CN 108085304 A CN108085304 A CN 108085304A
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- polyphenol oxidase
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0055—Oxidoreductases (1.) acting on diphenols and related substances as donors (1.10)
- C12N9/0057—Oxidoreductases (1.) acting on diphenols and related substances as donors (1.10) with oxygen as acceptor (1.10.3)
- C12N9/0059—Catechol oxidase (1.10.3.1), i.e. tyrosinase
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y110/00—Oxidoreductases acting on diphenols and related substances as donors (1.10)
- C12Y110/03—Oxidoreductases acting on diphenols and related substances as donors (1.10) with an oxygen as acceptor (1.10.3)
- C12Y110/03001—Catechol oxidase (1.10.3.1), i.e. tyrosinase
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2101/00—Agricultural use
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2109/00—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE pH regulation
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Agronomy & Crop Science (AREA)
- Molecular Biology (AREA)
- Materials Engineering (AREA)
- General Life Sciences & Earth Sciences (AREA)
- Soil Sciences (AREA)
- Mycology (AREA)
- Medicinal Chemistry (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Virology (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- Processing Of Solid Wastes (AREA)
- Soil Conditioners And Soil-Stabilizing Materials (AREA)
Abstract
The invention discloses a kind of high soil-repairing agents and its production method and application containing polyphenol oxidase, wherein described production method is to utilize stalk and kitchen garbage, it is equipped with inorganic salts, it is sterilized to make base-material, it will be inoculated into again with the plant endogenesis epiphyte seed liquor of liquid Czapek's medium shaking table culture on base-material, the temperature-variable fermentation culture in sterile room, the tunning of acquisition is the high soil-repairing agent containing polyphenol oxidase.The production of the soil-repairing agent of the present invention is not affected by environment, and with short production cycle, simple production process, production time and yield are easily controllable, and soil remediation effect is apparent.Concrete application may be employed directly make an addition to soil and impregnate seed, the method for impregnating plant roots or pouring makes it act on plant, method is simple, easy to operate, and dosage is few, it is at low cost and it is equal can achieve good results, belong to green agriculture.
Description
Technical field
The invention belongs to agriculture microbe technologies, and in particular to a kind of high soil-repairing agent containing polyphenol oxidase and
Its production method and application.
Background technology
Polyphenol oxidase (polyphenol oxidase, EC 1.10.3.1) is the oxidoreducing enzyme of a kind of cupric, is had
Extensive substrate, can not only a variety of aromatic compounds of catalysis oxidation, moreover it is possible to lignin degrading, many toxic aldehydes matters of removal
Toxicity, a variety of dyestuffs and its waste water decoloring can also be made, therefore in papermaking, environmental protection, food, medical and health, biological detection etc.
Field polyphenol oxidase has larger research and application value.Since polyphenol oxidase can also aoxidize many organic contaminations
Object, and particularly important effect is played in soil aromatic series organic matter humification process, therefore it is in terms of soil remediation
Application potential also gradually it is in widespread attention.
China's agricultural straw resource enriches, and annual output is occupied about at 700,000,000 tons or so first of the world according to statistics, and average
Every year still with 1251.2 ten thousand tons of speed steady-state growth, wherein corn, rice and wheat stalk yield is maximum, but straw utilization is imitated
Rate is relatively low.In recent years, when every summer and autumn crops are rush-harvested and rush-plant, often figure facilitates saving of labor to save trouble to peasant, usually pays stalk
One of torch.Open Burning Crop Residue is the rich and influential family of agricultural pollution, since stalk is mostly imperfect combustion under open-air atmosphere, is generated big
Measure the pollutant of harmful to human respiratory tract.The dense smog triggered is burned, makes a very bad impression to air environment, also be easy to cause
The communal facilitys such as electric wire, cable, communication line are damaged, and are triggered fire, are caused people and animals' injures and deaths and property loss, even resulting in can see
Degree declines, and Flights Delayed normally takes off, interferes highway normal operation repeatedly.Therefore, recycling China stalk resource
It develops eco-agriculture significant with sustainable agriculture.
China is time-honored food and drink big country, and the special eating habit of our people causes the big volume production of kitchen garbage
It is raw.It is counted according to urban construction department of the Ministry of Construction, China generated 3.9 ten thousand tons of kitchen garbages 3.9 hundred million urban population day.The rubbish of such enormous amount
It can not quickly be dissolved in city, all flow to the rural area of surrounding city, this brings white elephant to the environment in rural area, also gives
Illegal retailer feeds swill pig, it is hidden that this feeding manner also brings serious food security with opportunity with kitchen garbage
Suffer from.
The grease weight of kitchen garbage, fat content are high, putrefactive phenomenon, micro- life can occur mostly in storage, transportational process
Object, bacterium amount reproduction.Wherein contain the pathogenic microorganism of a large amount of infectious diseases common to human beings and animals, not only easily cause zoogenetic infection husky
Door bacterium disease, colibacillosis etc. more than 10 plants infectious disease, and pathogenic microorganism colonizes in the internal procreation of pig, into food chain
Afterwards, human infection disease (such as hepatitis) can be caused, " the recessive bomb " for being known as communicable disease is caused to be broken out by sanitationist.
Meanwhile the leftovers of starch etc are also easily generated under the action of bacterium and gone mouldy.Contained by putrid and deteriorated swill
Various toxin can cause endotoxin contamination to domesticated animal, and such as the Aflatoxin that aspergillus flavus generates, the kojic acid that aspergillus oryzae generates is all
With very strong carcinogenicity, the diseases such as intestinal cancer, stomach cancer, liver cancer, kidney, breast cancer can be caused.
The harmless treatment of kitchen garbage is imperative, this problem has also caused the great attention of government.By
In kitchen garbage water content is high, fat content is high, salinity weight, calorific value is low the features such as, with landfill, burn, compost the methods of handle
Effect is undesirable.Expensive processing cost simultaneously also virtually adds the difficulty of kitchen garbage improvement.Therefore, in kitchen
On the governing problem of rubbish, preferable method be not only solve rubbish it is innoxious, reduce environmental pollution, ensure that food security is defended
It is raw, while more resource reutilization, maximum limit are carried out with the means of science using kitchen garbage as a kind of available resource
The waste of the reduction resource of degree could also solve food-safety problem and environmental problem from source.
Some plants, plant in same field for years, are often present with yellow, runt seedling, even large stretch of plant death,
Severely impact the raising of yield and quality.This phenomenon is referred to as continuous cropping obstacle on cultivation.Cause plant continuous cropping obstacle
The reason for mainly have that soil nutrient is unbalance, structure of soil microbial community changes, phytotoxic substance increases and soil physics
Learn property change etc..Guo Lanping etc. by the identification of rhizoma atractylodis rhizome and Rhizosphere Soil water extract bioactivity research and allelochemical,
Prompting Auto toxicity results in cultivation rhizoma atractylodis continuous cropping obstacle.Simultaneously by being ground to cultivation rhizoma atractylodis rhizosphere soil microorganism variation
Study carefully, it is believed that the generation of rhizoma atractylodis pest and disease damage is cultivated for many years, it may be with the change of its rhizosphere area soil microbe quantity and structure of community
Caused continuous cropping obstacle is related.
Endophyte of plant refers to be present in health plant histoorgan, does not form an apparent pathogenic quasi-microorganism.
Current study show that there is the presence of endophyte in all plants being investigated.Endophyte of plant research is to improve seed hair
Bud rate accelerates medicinal material growth, improves resistance, promotes active principle accumulation that medical material quanlity is maintained to solve the one of continuous cropping obstacle
A promising direction.The plant host for infecting endophyte often has fast growing, degeneration-resistant border, disease resistance, anti-animal injury
Etc. advantages, the plant than being uninfected by endophyte have more struggle for existence power.Mo Wenping etc. has found salt Promoting bacteria Rs1524 processing cottons
Afterwards, germination percentage improves 11.8% than control, and leaf little Mei etc. has found the hair of the green vegetables handled through 102 bacterium dilutions, tomato seeds
Bud rate and root long significantly improve, and the researchs such as high beautiful luxuriant growth show that endophyte can promote rye grass drought resisting, degeneration-resistant border, Chen Xiaomei etc.
Under isolated culture condition, it was found that endogenetic fungus can influence the growth of HERBA DENDROBII and total biology with HERBA DENDROBII symbiosis culture
The content of alkali improves the content of polysaccharide.Some endogenetic fungus have suppression harmful bacteria certain capacity of decomposition, therefore utilize Nei Shengzhen
Bacterium can solve or alleviate Auto toxicity.
Since continuous cropping obstacle leads to problems such as the percentage of seedgermination of crop low, slow-growing, easily sick, the country always cannot
It solves very well.We, which test to show to select in plant endogenesis epiphyte access stalk and the culture medium of kitchen garbage mixing, is made soil
Earth renovation agent will be an effective way, be overcome with the soil-repairing agent of high producing enzyme or eliminated continuous cropping obstacle unmanned report at present
Road, the soil remediation agent and process for producing same is without patent.
The content of the invention
The purpose of the present invention is being directed to drawbacks described above of the prior art, provide one kind can by comprehensively utilize stalk with
And kitchen garbage realizes extensive raise crop, overcomes or eliminate the high soil-repairing agent containing polyphenol oxidase of continuous cropping obstacle,
And provide the preparation method and concrete application of the soil-repairing agent.
Technical solution：A kind of production method of high soil-repairing agent containing polyphenol oxidase of the present invention, including with
Lower step：
(1) seed liquor culture：Plant endogenesis epiphyte strain is taken, after culture activation, mycelia shaking table culture is scraped, obtains seed
Liquid；
(2) base-material prepares：Stalk and kitchen garbage are taken, is pulverized and mixed, it is 20 to adjust C/N:1-40:1, then add inorganic
Salt, after mixing sterilizing obtain base-material；
(3) fermented and cultured：Seed liquor obtained by step (1) is inoculated into obtained by step (2) on base-material, inoculum concentration 4%-
8%, base-material is then placed in Fluctuation temperature culture in sterile room, it is the high soil remediation containing polyphenol oxidase to obtain tunning
Agent.
In step (1), the plant endogenesis epiphyte of the energy high yield polyphenol oxidase refers to be present in health plant tissue
It in organ, is not formed apparent pathogenic, can efficiently produce a kind of eukaryotic microorganisms of polyphenol oxidase.Existing energy high yield polyphenol
Oxidizing ferment, and the method that the plant endogenesis epiphyte of plant growth is promoted to could be used for the present invention, the present invention recommend following strain：Ball
Cupreum, intend mould pestalotia bacteria, Alternaria, Cunninghamella sp, mould, Phoma sp, sickle-like bacteria, hole ball spore, bulbil bacterium,
Branch acremonium, angle basidiomycetes, rhizoctonia, bulbil bacterium, small single head spore, cupreum, shell roundlet spore, stick disk spore category, vesicle shell category,
Phomopsis.
Specific available plant endogenesis epiphyte is listed as follows：
Further, in step (1), the plant endogenesis epiphyte strain is selected from chaetomium globosum (BNCC150438) or intends disk
Pestalotia bacteria (BNCC151148).
Step (1) concrete operations are to take out strain from 4 DEG C of refrigerators, be transferred to PDA inclined-planes, 25 DEG C grow 7 days, pour into nothing
Bacterium water scrapes mycelia, the liquid Czapek's medium transferred into cane sugar content for 0.5-1.5g/L, 22-30 DEG C, 160-200r/min
Shaking table culture 3-7 days, obtains seed liquor.
In step (2), preferred C/N is 20:1-30:1.
In step (2), the stalk is selected from one or both of wheat stalk and rice straw, can also use other
It is suitable as the agricultural wastes substituted.
In step (2), the inorganic salts additive amount is 0.5-1.5g/L.
In step (2), sterilizing refers to through 121 DEG C, 20min of high-pressure sterilizing pot (steam boiler), 100kpa high pressure sterilizations.
In step (3), preferably inoculum concentration is 6%-8%.
In step (3), the Fluctuation temperature culture refers to temperature setting for 15-30 DEG C of daytime, and in 10-20 DEG C of evening, one co-cultures
14-20 days.
It is also within the scope of the present invention that the high soil-repairing agent containing polyphenol oxidase of gained is prepared according to the above method.
Further, contained polyphenol oxidase reaches 300-600U/ in the high soil-repairing agent containing polyphenol oxidase
g.Wherein, the enzyme activity of polyphenol oxidase is 300U.
Above-mentioned soil-repairing agent is also within the scope of the present invention for the application of soil remediation.
Above-mentioned tunning can directly make an addition to soil as soil-repairing agent or be uniformly mixing to obtain Inoculant,
It can be used for seed soaking, pouring root or leaching root to use.
Above-mentioned tunning directly makes an addition to soil and can be used for overcoming or eliminating continuous cropping obstacle, our experimental result table
The soil-repairing agent of bright high yield polyphenol oxidase can pole significantly increase the germination percentage of Atractylis lancea seed, while promote crop
Growth improves resistance, disease-resistant insect resistance capacity, more accumulation of stimulating activity substance.It is considered that selected endophyte soil
Earth renovation agent can be by improving soil physical chemistry situation, culture fertility, improving soil nutrient utilization rate, decomposition suppression harmful bacteria, drive
Keep away the effect of subterranean pest-insect, so as to overcoming or eliminate continuous cropping obstacle.Above-mentioned described continuous cropping obstacle refers to some plants, continuous more
Year plants in same field, is often present with yellow, runt seedling, even large stretch of plant death, severely impacts yield and quality
It improves.
The present invention is the method for comprehensive utilization to agricultural wastes and kitchen garbage, and stalk and food and drink rubbish from cooking are mixed
It closes, using the high carbon elements of stalk, the characteristics of the high Nitrogen element of food and drink rubbish from cooking and moisture, adjusts C/N, access Nei Shengzhen
Bacterium, selected plant endogenesis epiphyte pole considerably improve polyphenol oxidase production of enzyme.
Gained tunning is used for soil remediation, can be directly added into soil, shifted, absorbed using microbial technique,
Pollutant in degradation and conversion soil, adjusts soil pH value, eliminates soil hardening, inhibits soil-borne disease, reaches improvement soil
The purpose of structure makes soil restoration to ecosystem structure；The method for impregnating seed, immersion plant roots or pouring can also be used to make
It acts on plant, adjusts the carbon nitrogen balance of plant rhizosphere, improves the micro-ecological environment of plant growth, improves percentage of seedgermination,
Promote the growth of crop, improve resistance, disease-resistant insect resistance capacity, so as to effectively overcome or eliminate continuous cropping obstacle.
The soil-repairing agent of the present invention can also effectively reduce the use of pesticide etc., so as to reduce agricultural environment pollution protection
The effect of ecology plays the role of the effect that repellent subterranean pest-insect resists pathogen.
Substantial amounts of viable bacteria group is colonized in crop root in the present invention simultaneously, mushrooms out crop root, deep bundle, can
Yield and quality, the quality of agricultural product are improved, is the preferable fertilizer of organic agriculture, there is important answer in large-scale industrialized production
With value, there are bright prospects in agricultural sustainable development.
Advantageous effect：The production of the soil-repairing agent of the present invention is not affected by environment, with short production cycle, production technology letter
Single, production time and yield are easily controllable, and soil remediation effect is apparent.Concrete application may be employed directly make an addition to soil and
The method for impregnate seed, impregnating plant roots or pouring makes it act on plant, and method is simple, easy to operate, and dosage is few, it is at low cost and
It can achieve good results.In addition, polyphenol oxidase can aoxidize many organic pollutions, and in soil organism humification
Particularly important effect is played in the process, and one long period still has and plays a role, therefore this soil-repairing agent acts on
Persistently, the use of the chemical substance of the pollution environment such as pesticide can be effectively reduced, so as to reduce agricultural environment pollution protecting ecology
Effect, belong to green agriculture.
Description of the drawings
Fig. 1 is the flow chart of production method of the present invention.
Specific embodiment
Used term in the present invention, it is unless otherwise specified, generally usual with those of ordinary skill in the art
The meaning of understanding.
With reference to specific embodiment, description is of the invention in further detail.It should be understood that these embodiments are intended merely to
It illustrates the present invention rather than limits the scope of the invention in any form.
Below in an example, the various processes and method not being described in detail are conventional methods as known in the art.
The source of agents useful for same, trade name and it is necessary to list its constituent person are indicated, thereafter phase used on the first appearance
With reagent unless otherwise specified, it is identical with the content indicated for the first time.
Embodiment 1
Chaetomium globosum (Chaetomium globosum) is taken out from 4 DEG C of refrigerators, is transferred to PDA inclined-planes, and 25 DEG C grow 7 days,
Sterile water is poured into, mycelia is scraped, transfers into liquid Czapek's medium, 25 DEG C, 180r/min shaking table cultures 3 days obtain a large amount of bacterium
Liquid.Together with kitchen garbage after stalk crushing, it is 20 to adjust C/N:1, it is equipped with 1.2g/L NaNO3, it is put into high-pressure sterilizing pot (steam
Boiler) high pressure sterilization, solid fermentation culture medium is made.Bacterium solution is accessed into solid fermentation culture medium, inoculum concentration 6%, Jiang Qidui
25cm high in sterile room, with incubator Fluctuation temperature culture, stimulates producing enzyme.Temperature setting is 25 DEG C of daytime, 20 DEG C at night, altogether
Culture 15 days.Product is obtained, contained polyphenol oxidase reaches 300-600U/ in soil-repairing agent of the gained height containing polyphenol oxidase
g.Wherein, the enzyme activity of polyphenol oxidase is 300U.For being applied directly in arable farming soil.
It is reflected in table 1 and adds endogenetic fungus chaetomium globosum, sterile hair in chrysanthemum sequential cropping ground respectively within 3 years in continuous plantation
After ferment dilution and embodiment 1 prepare gained containing bacterium fermentation dilution, chaetomium globosum and its fermentation dilution liquid energy improve continuous cropping
Chrysanthemum yield and quality in chrysanthemum ground.Each processing 50 florists chrysanthemum seedlings of cuttage.Processing 1：Apply sterile water (applied amount：200mL without
Bacterium water/chrysanthemum)；Processing 2：Apply zymotic fluid (applied amount：Fermentation dilutions of the 200mL without chaetomium globosum/chrysanthemum)；Processing
3：Apply chaetomium globosum and sterile water (applied amount：Weight in wet base bacterium/chrysanthemum and 200mL sterile waters/chrysanthemum after 5g is filtered)；Processing
4：Apply chaetomium globosum and zymotic fluid (applied amount：5g filter after the fermentation dilution of weight in wet base bacterium and 200mL without chaetomium globosum/
Chrysanthemum).More than 4 processing, per 60d reprocessing once, in entire Dendranthema Growth phase coprocessing 3 times.
Influence of 1 different disposal of table to chrysanthemum yield and economical character
Embodiment 2
Angle basidiomycetes (Ceratobasidium stevensii) takes out from 4 DEG C of refrigerators, is transferred to PDA inclined-planes, 25 DEG C of lifes
It is 7 days long, sterile water is poured into, mycelia is scraped, transfers into liquid Czapek's medium, 30 DEG C, 200r/min shaking table cultures 3 days obtain
A large amount of bacterium solutions.Together with kitchen garbage after stalk crushing, it is 30 to adjust C/N:1, it is equipped with 0.5g/L NaNO3, it is put into high-pressure sterilizing pot
(steam boiler) high pressure sterilization, is made solid fermentation culture medium.Bacterium solution is accessed into solid fermentation culture medium, inoculum concentration 4% will
Its heap 25cm high in sterile room, with incubator Fluctuation temperature culture, stimulates producing enzyme.Temperature setting is 30 DEG C of daytime, 20 DEG C at night,
One co-cultures 14 days.Product is obtained, contained polyphenol oxidase reaches 300- in soil-repairing agent of the gained height containing polyphenol oxidase
600U/g.Wherein, the enzyme activity of polyphenol oxidase is 300U.Zymotic fluid is used as immersion inoculation during budding.
In table 2, soil collection is in Yongzhou City, Hubei Province Hengyang Red Soil Experimental Station, Chinese Academy of Agricultural Sciences Meng public affairs mountain demonstration base
The red soil (5-20cm) of the Quaternary Period red clay development on ground, it is continuous to plant watermelon 3 years.After applying angle basidiomycetes, polyphenol oxidase
Activity increase significantly.Experiment totally 3 processing, 6 repetitions.Processing is respectively control (A)：It is not added with mycelia；B：Application is gone out
Mycelia living；C：Apply mycelia living.Seed carries out (55 DEG C) seed soaking of lukewarm water prior to seeding, then natural cooling 2h is soaked respectively
Kind 4h, A distilled water 50mL in 3 processing of A, B, C；The angle basidiomycetes mycelia 2g+48mL water of B inactivations；The angle basidiomycetes that C lives
Mycelia 2g+48mL water.Normal irrigation management, June 1, transplanting watermelon seedling is into experiment basin alms bowl.Basin alms bowl diameter 28cm is tested, it is high
28cm, per basin soil 15kg, respectively using urea 4g, potassium dihydrogen phosphate 4g, borax 0.55g, alkaline residue 60g, organic fertilizer 500g.This
Outside, B, C processing apply the wet mycelia 20g of B6 of the wet mycelia of angle basidiomycetes and work of inactivation respectively.Normal field management.July 10
(sitting the melon phase), B, C processing apply the wet mycelia 20g of B6 of the wet mycelia of angle basidiomycetes and work of inactivation respectively.July 29, picking west
Melon.It before watermelon is transplanted, in watermelon florescence, sits melon phase and maturity period and fetches earth, measure the vigor of soil polyphenol oxidase.
Table 2 applies influence of the angle basidiomycetes to watermelon soil polyphenol oxidase
Embodiment 3
Phomopsis (Phomopsis sp.) takes out from 4 DEG C of refrigerators, is transferred to PDA inclined-planes, and 25 DEG C grow 7 days, pour into nothing
Bacterium water scrapes mycelia, transfers into liquid Czapek's medium, 22 DEG C, 160r/min shaking table cultures 3 days obtain a large amount of bacterium solutions.Stalk
Together with kitchen garbage after crushing, it is 40 to adjust C/N:1, it is equipped with 1.5g/L NaNO3, it is put into high-pressure sterilizing pot (steam boiler) height
Pressure sterilizing, is made solid fermentation culture medium.Bacterium solution is accessed into solid fermentation culture medium, inoculum concentration 8%, by its heap 25cm high,
In sterile room, with incubator Fluctuation temperature culture, producing enzyme is stimulated.Temperature setting is 15 DEG C of daytime, 10 DEG C at night, and one co-cultures 20
My god.Fermentation ends add 0.1% agar or gelatin to be stirred evenly as thickener.Product is obtained, gained height is containing polyphenol oxidase
Contained polyphenol oxidase reaches 300-600U/g in soil-repairing agent.Wherein, the enzyme activity of polyphenol oxidase is 300U.For crop
Transplanting leaching root.
In table 3 and table 4, potting soil is derived from red soil ecology experiment station of the Yingtan Chinese Academy of Sciences of Jiangxi Province, is red clay
The red soil (5-20cm) of development, Peanut continuous cropping for many years, after applying Phomopsis B3, the activity of polyphenol oxidase, which has, significantly to be carried
Height, and peanut agronomic trait increases significantly compared to control.3 processing：It compares (CK)；Endogenetic fungus B3 is added in soil
(B3+)；Addition inactivation endogenetic fungus B3 (B3-) in soil.It tests all each time sampling points of processing and is all provided with 3 repetitions.With
It sows peanut immediately afterwards, per two, basin, ensures sunshine and moisture.Distinguish (the 0th day), peanut germination period the (the 10th prior to seeding
My god), seedling stage (the 30th day), florescence (the 60th day), fruiting period (the 90th day) and (the 110th day) maturity period be measured by sampling.
Table 3 applies influence of the Phomopsis to polyphenol oxidase activity in soil
The letter that each column numerical value marks below represents the significant difference (P between different numerical value<0.05)
Influence of 4 endogenetic fungus of table to peanut agronomic trait
Letter a, b, c represent P respectively in table<0.05 significance level of difference.
Claims (10)
1. a kind of production method of the high soil-repairing agent containing polyphenol oxidase, which is characterized in that comprise the following steps：
(1) seed liquor culture：Plant endogenesis epiphyte strain is taken, after culture activation, mycelia shaking table culture is scraped, obtains seed liquor；
(2) base-material prepares：Stalk and kitchen garbage are taken, is pulverized and mixed, it is 20 to adjust C/N:1-40:1, then inorganic salts are added, it mixes
Sterilizing obtains base-material after closing uniformly；
(3) fermented and cultured：Seed liquor obtained by step (1) is inoculated into obtained by step (2) on base-material, inoculum concentration 4%-8%, so
Base-material is placed in Fluctuation temperature culture in sterile room afterwards, it is the high soil-repairing agent containing polyphenol oxidase to obtain tunning.
2. the production method of soil-repairing agent of the height containing polyphenol oxidase according to claim 1, which is characterized in that step
(1) in, the plant endogenesis epiphyte strain be selected from chaetomium globosum, intend pestalotia bacteria, Alternaria, Cunninghamella sp, mould,
Phoma sp, sickle-like bacteria, hole ball spore are mould, bulbil bacterium, branch acremonium, angle basidiomycetes, rhizoctonia, bulbil bacterium, small single head spore, hair
One or more in shell bacterium, shell roundlet spore, stick disk spore category, vesicle shell category, Phomopsis.
3. the production method of soil-repairing agent of the height containing polyphenol oxidase according to claim 1, which is characterized in that step
(1) in, the plant endogenesis epiphyte strain is selected from chaetomium globosum or intends pestalotia bacteria.
4. the production method of soil-repairing agent of the height containing polyphenol oxidase according to claim 1, which is characterized in that step
(2) in, C/N is 20 in the base-material:1-30:1.
5. the production method of soil-repairing agent of the height containing polyphenol oxidase according to claim 1, which is characterized in that step
(2) in, the inorganic salts additive amount is 0.5-1.5g/L.
6. the production method of soil-repairing agent of the height containing polyphenol oxidase according to claim 1, which is characterized in that step
(3) in, inoculum concentration 6%-8%.
7. the production method of soil-repairing agent of the height containing polyphenol oxidase according to claim 1, which is characterized in that step
(3) in, the Fluctuation temperature culture refers to temperature setting for 15-30 DEG C of daytime, and 10-20 DEG C of evening cultivates 14-20 days.
8. any production method prepares the high soil-repairing agent containing polyphenol oxidase of gained in claim 1-7.
9. the high soil-repairing agent containing polyphenol oxidase according to claim 8, which is characterized in that described high containing polyphenol oxidase
Contained polyphenol oxidase reaches 300-600 units/grams in the soil-repairing agent of enzyme.
10. soil-repairing agent described in claim 9 is used for the application of soil remediation.
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Application publication date: 20180529 |