CN108164322A - A kind of preparation method of soil sterilants - Google Patents
A kind of preparation method of soil sterilants Download PDFInfo
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- CN108164322A CN108164322A CN201711257114.4A CN201711257114A CN108164322A CN 108164322 A CN108164322 A CN 108164322A CN 201711257114 A CN201711257114 A CN 201711257114A CN 108164322 A CN108164322 A CN 108164322A
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
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- 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
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/08—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing solids as carriers or diluents
- A01N25/10—Macromolecular compounds
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- 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
- A01N63/10—Animals; Substances produced thereby or obtained therefrom
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- 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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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- A—HUMAN NECESSITIES
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- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/40—Liliopsida [monocotyledons]
- A01N65/42—Aloeaceae [Aloe family] or Liliaceae [Lily family], e.g. aloe, veratrum, onion, garlic or chives
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F5/00—Fertilisers from distillery wastes, molasses, vinasses, sugar plant or similar wastes or residues, e.g. from waste originating from industrial processing of raw material of agricultural origin or derived products thereof
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Abstract
The invention discloses a kind of preparation methods of soil sterilants, belong to disinfectant field.The present invention extracts a kind of antibacterial material from fly body, the special mode of this antibacterial material inhibits the synthesis of the large biological molecules such as fungal Internal chitin, so as to influence the Energy distribution of somatic cells film surface, along with rhizoma atractylodis, grass-leaved sweetflag, folium artemisiae argyi, due to rhizoma atractylodis, grass-leaved sweetflag, folium artemisiae argyi all has certain bactericidal effect, package release is carried out using sodium alginate, so as to extend the time limit of disinfectant antibacterial, and sodium alginate has film forming, it is adsorbed in microbial cell surface, form a floor height molecular film, nutriment is prevented to intracellular transport, so as to play sterilization and bacteriostasis effect.The present invention is solved current soil sterilants and mainly soil is sprayed using chemical agent or stifling etc., although the biologies such as the bacterium in soil can be killed, medicament residue, and the problem of causing environmental pollution dangerous, difficult to degrade to plant.
Description
Technical field
The invention belongs to disinfectant fields, and in particular to a kind of preparation method of soil sterilants.
Background technology
The pathogen of soil-borne disease is generally more host micro-organisms, can infect several or even hundreds of crops and miscellaneous
Grass has very strong viability and infecting potential, regards as being most difficult to one of the disease of prevention by plant kingdom.In agriculture and forestry, soil
Quality be to restrict the key factor of this field development, in agriculture and forestry the soil of raise crop be by clay, sand, organic matter,
The compositions such as water, air, clay account for about 40%, and water and air respectively accounts for 30%, but also dwells give birth to bacterium, fungi, virus in the soil
And the harmful organisms such as nematode, they influence the growth and development of plant, so as to reduce yield and quality, allow peasant by great warp
Ji loss.The main place that soil is the main media of propagation pest and disease damage and disease pest is bred, many germs, worm's ovum and pest
All existence or overwintering in the soil.China's vegetable cultivation area is more than 19,700,000 hectares, and wherein Installation Vegetable Cultivation area is up to 250
More ten thousand hectares, account for more than the 80% of world's facilities vegetable gross area.Compared with developed countries, the intensive vegetable filed land use in China is strong
Degree is high, and usage frequency is high, and multiple crop index is high, and planting density is big.It is intensive vegetable filed to recycle, substantially without time out.Facility dish
Field high temperature, high humidity, closing or semiclosed environment cause soil-borne disease serious, and pest and disease damage occurs that frequently, victual can be reduced
Yield and quality, the underproduction even total crop failure when serious.
Current soil sterilants is mainly chemosterilant, includes the use of formaldehyde, Cosan, pulverized limestone, carbendazim, generation
The medicaments such as gloomy zinc, pentachloronitrobenzene, Urbasulf, fenaminosulf, ferrous sulfate, metalaxyl, Mancozeb are mixed in the soil to soil
Earth is sprayed or stifling etc., and medicament directly acts on soil, and effect is good, using universal, although using chemical agent energy
Kill the biologies such as bacterium, fungi, nematode in soil, but have after certain medicament residue it is dangerous to plant and these
Chemical agent is difficult often that degradation pollutes the environment.Therefore, a kind of drug effect height is produced and to the soil of eco-friendly
Earth disinfectant has the very big market demand.
Invention content
The technical problems to be solved by the invention：Mainly soil is carried out using chemical agent for current soil sterilants
Spray or fumigate etc., although the biologies such as the bacterium in soil can be killed, medicament residue is dangerous, difficult to degrade to plant to cause
The problem of environmental pollution, provides a kind of preparation method of soil sterilants.
In order to solve the above technical problems, the present invention is using technical solution as described below：
A kind of preparation method of soil sterilants, which is characterized in that the preparation method includes the following steps：
（1）Fly is put into the water that temperature is 0 ~ 4 DEG C and is ground, the benzyl sulfuryl fluoride of water quality 0.01 ~ 0.02% is added in, grinds
25 ~ 30min is ground, lapping liquid is obtained, lapping liquid, phosphate buffer is mixed, 25 ~ 30min is stood at 0 ~ 4 DEG C of temperature, and
Carry out 25 ~ 30min of centrifugation, obtain supernatant, supernatant is heated, temperature to 90 ~ 98 DEG C, 3 ~ 4min of time, then carry out from
25 ~ 30min of the heart, takes supernatant, and supernatant is moved into carboxymethyl cellulose chromatographic column carries out cation-exchange chromatography, then use phosphoric acid
Buffer solution, sodium chloride solution are eluted, and collect eluent；
（2）It counts in parts by weight, 3 ~ 4 parts of kaffir lily leaves, 3 ~ 4 parts of sansevieria trifasciata prain leaves, 1 ~ 2 part of grass-leaved sweetflag is taken to be mixed, is dried, then
It is put into pulverizer and crushes, be sieved, obtain sieving, sodium alginate, water are mixed, and is stirred, add alginic acid
The sieving of sodium quality 10 ~ 15%, is stirred, and obtains stirring mixture, stirring mixture is dried at 50 ~ 60 DEG C of temperature, powder
Broken, sieving obtains sieving a, mixes, and be stirred 20 ~ 30min eluent, sieving a, obtain mixed liquor, spare；
（3）Soil sample, sterile water in flower nursery are mixed, 25 ~ 30 min of shaken cultivation at 30 ~ 40 DEG C of temperature stands 1 ~ 2h,
Supernatant is taken, 50 ~ 60% water dilution is added in into supernatant, in 70 ~ 80 DEG C of 10 ~ 15min of water-bath of temperature, obtains dilution, it will be dilute
It releases liquid to be seeded in solid medium with oese, 1 ~ 2d is cultivated in 20 ~ 30 DEG C of insulating boxs of temperature, picking solid medium
Bacterium colony, and be seeded to the flat lining out of beef extract-peptone and isolate and purify, the bacterium colony of bacterium diameter maximum is selected, by the bacterium of bacterium diameter maximum
Fall, fluid nutrient medium is mixed, in 25 ~ 30 DEG C of temperature, 120 ~ 160r/min, 15 ~ 18h of shaking table culture obtain culture solution；
（4）Activated sludge, molasses, water are mixed, high-temperature sterilization, culture solution is added in, in 25 ~ 30 DEG C of temperature, 160 ~ 180r/
Ferment 1 ~ 2d under the conditions of min, adds turf, after mixing, dry at 40 ~ 50 DEG C of temperature, and crushes, and must crush
Object, by step（2）Spare mixed liquor, crushed material are mixed, and are stirred 10 ~ 15min to get soil sterilants.
The step（1）Middle fly, the mass ratio 1 for the water that temperature is 0 ~ 4 DEG C：5 ~ 6, lapping liquid, phosphate buffer matter
Measure ratio 1：4~5.
The step（1）Middle phosphate buffer is in mass ratio 1：2：5 ~ 6, by sodium dihydrogen phosphate, disodium hydrogen phosphate, water into
Row mixing is to get phosphate buffer.
The step（2）The mass ratio 1 of middle chitosan, the water that temperature is 80 ~ 90 DEG C：3 ~ 4, eluent, sieving a matter
Measure ratio 10 ~ 15：1~2.
The step（3）The mass ratio 1 of soil sample in middle flower nursery, sterile water：10 ~ 15, the bacterium colony of bacterium diameter maximum, Liquid Culture
The mass ratio 1 of base：5~6.
The step（3）Middle solid medium is to count in parts by weight, take 40 ~ 50 parts of water, 10 ~ 15 parts of agar, 10 ~ 15 parts
Mannitol, 5 ~ 7 parts of calcium carbonate, 0.2 ~ 0.4 part of potassium dihydrogen phosphate, 0.2 ~ 0.4 part of magnesium sulfate, 0.2 ~ 0.4 part of sodium chloride, 0.1 ~
0.2 part of calcium sulfate is mixed, and high-temperature sterilization is to get solid medium.
The step（3）Middle beef extract-peptone tablet is to count in parts by weight, take 50 ~ 60 parts of water, 15 ~ 25 parts of agar,
10 ~ 15 parts of peptones, 5 ~ 7 parts of sodium chloride, 3 ~ 4 portions of beef extracts are mixed, and high-temperature sterilization is to get beef extract-peptone tablet.
The step（3）Middle fluid nutrient medium is to count in parts by weight, take 40 ~ 50 parts of water, 10 ~ 15 portions of mannitol, 5 ~ 7 parts
Calcium carbonate, 0.2 ~ 0.4 part of potassium dihydrogen phosphate, 0.2 ~ 0.4 part of magnesium sulfate, 0.2 ~ 0.4 part of sodium chloride, 0.1 ~ 0.2 part of calcium sulfate into
Row mixing, high-temperature sterilization is to get fluid nutrient medium.
The step（4）In count in parts by weight, take 40 ~ 50 parts of water, 40 ~ 50 parts of activated sludge, 10 ~ 15 parts of molasses, 5 ~ 6
Part culture solution, 3 ~ 4 parts of turfs, step（2）Spare mixed liquor, the mass ratio 30 ~ 40 of crushed material：5~6.
Compared with other methods, advantageous effects are the present invention：
（1）The present invention extracts a kind of antibacterial material from fly body, and the special mode of this antibacterial material inhibits fungal Internal several
The synthesis of the large biological molecules such as fourth matter so as to influence the Energy distribution of somatic cells film surface, causes somatic cells film to go to pole
Change, ion and metabolite leakage, and other basic functions forfeiture such as respiration causes, along with rhizoma atractylodis, grass-leaved sweetflag, Chinese mugwort
Since rhizoma atractylodis, grass-leaved sweetflag, folium artemisiae argyi all have certain bactericidal effect, package release is carried out using sodium alginate for leaf, so as to
Extend the time limit of disinfectant antibacterial, and sodium alginate has film forming, absorption is in microbial cell surface, formation one floor height point
Sub- film prevents nutriment to intracellular transport, so as to play sterilization and bacteriostasis effect, what several Fungicidal substance mixing distributed
Smog sterilizing power is strong, and so as to further improve the bactericidal effect of disinfectant, and rhizoma atractylodis have drying damp and strengthening spleen, the work(of expelling wind and clearing away cold
Effect, folium artemisiae argyi have effects that eliminating cold to stop pain, and the two combines, has the function of cough-relieving, eliminating the phlegm, relievings asthma, can prevent influenza,
Therefore soil fungicides of the present invention can carry out sterilizing in the case of presence of people completely.
（2）The present invention also added in disinfectant it is a kind of using activated sludge for substrate progress mixed fermentation compound bacteria is made
Fertilizer not only takes full advantage of discarded activated sludge resource, while certain nutrient can be provided for soil, so as to improve soil
Earth fertility, therefore soil sterilants produced by the present invention has splendid Disinfection Effect, can improve soil fertility, promotes soil matter
Amount, and soil sterilants will not damage to environment and human body without chemical agent.
Specific embodiment
Phosphate buffer is in mass ratio 1：2：5 ~ 6, sodium dihydrogen phosphate, disodium hydrogen phosphate, water are mixed to get phosphorus
Acid buffer.
Solid medium is to count in parts by weight, take 40 ~ 50 parts of water, 10 ~ 15 parts of agar, 10 ~ 15 portions of mannitol, 5 ~ 7 parts
Calcium carbonate, 0.2 ~ 0.4 part of potassium dihydrogen phosphate, 0.2 ~ 0.4 part of magnesium sulfate, 0.2 ~ 0.4 part of sodium chloride, 0.1 ~ 0.2 part of calcium sulfate into
Row mixing, high-temperature sterilization is to get solid medium.
Beef extract-peptone tablet is to count in parts by weight, take 50 ~ 60 parts of water, 15 ~ 25 parts of agar, 10 ~ 15 parts of peptones,
5 ~ 7 parts of sodium chloride, 3 ~ 4 portions of beef extracts are mixed, and high-temperature sterilization is to get beef extract-peptone tablet.
Fluid nutrient medium is to count in parts by weight, take 40 ~ 50 parts of water, 10 ~ 15 portions of mannitol, 5 ~ 7 parts of calcium carbonate, 0.2 ~
0.4 part of potassium dihydrogen phosphate, 0.2 ~ 0.4 part of magnesium sulfate, 0.2 ~ 0.4 part of sodium chloride, 0.1 ~ 0.2 part of calcium sulfate are mixed, high temperature
Sterilizing is to get fluid nutrient medium.
A kind of preparation method of soil sterilants, the preparation method include the following steps：
（1）In mass ratio 1：5 ~ 6, fly is put into the water that temperature is 0 ~ 4 DEG C and is ground, adds in water quality 0.01 ~ 0.02%
Benzyl sulfuryl fluoride, grind 25 ~ 30min, obtain lapping liquid, in mass ratio 1：4 ~ 5, lapping liquid, phosphate buffer are mixed,
25 ~ 30min is stood at 0 ~ 4 DEG C of temperature, and 25 ~ 30min of centrifugation is carried out with 1000 ~ 2000r/min, supernatant is obtained, by supernatant
It is heated, temperature carries out 25 ~ 30min of centrifugation to 90 ~ 98 DEG C, 3 ~ 4min of time, then with 1000 ~ 2000r/min, takes supernatant
Supernatant is moved into carboxymethyl cellulose chromatographic column and carries out cation-exchange chromatography by liquid, then with phosphate buffer, sodium chloride solution
It is eluted, collects eluent；
（2）It counts in parts by weight, 3 ~ 4 parts of rhizoma atractylodis, 3 ~ 4 parts of grass-leaved sweetflags, 1 ~ 2 portion of folium artemisiae argyi is taken to be mixed, dries, places into crushing
It is crushed in machine, crosses 70 ~ 80 mesh sieve, obtain sieving, in mass ratio 1：3 ~ 4, sodium alginate, water are mixed, and with 200 ~
300r/min is stirred, and is added the sieving of sodium alginate quality 10 ~ 15%, is stirred with 150 ~ 200r/min, must be stirred
Mixture is mixed, stirring mixture at 50 ~ 60 DEG C of temperature is dried, is crushed, 120 ~ 140 mesh sieve is crossed, sieving a is obtained, by quality
Than 10 ~ 15：1 ~ 2, eluent, sieving a are mixed, and be stirred 20 ~ 30min with 200 ~ 300r/min, must be mixed
Liquid, it is spare；
（3）In mass ratio 1：10 ~ 15, soil sample, sterile water in flower nursery are mixed, at 30 ~ 40 DEG C of temperature
25 ~ 30 min of shaken cultivation stands 1 ~ 2h, takes supernatant, 50 ~ 60% water dilution is added in into supernatant, in temperature 70
~ 80 DEG C of 10 ~ 15min of water-bath, obtain dilution, and dilution is seeded to oese in solid medium, in 20 ~ 30 DEG C of perseverances of temperature
1 ~ 2d, the bacterium colony of picking solid medium are cultivated in incubator, and is seeded to the flat lining out of beef extract-peptone and isolates and purifies, is chosen
Select the bacterium colony of bacterium diameter maximum, in mass ratio 1：5 ~ 6, the bacterium colony of bacterium diameter maximum, fluid nutrient medium are mixed, temperature 25 ~
30 DEG C, 120 ~ 160r/min, 15 ~ 18h of shaking table culture obtain culture solution；
（4）Count in parts by weight, take 40 ~ 50 parts of water, 40 ~ 50 parts of activated sludge, 10 ~ 15 parts of molasses, 5 ~ 6 parts of culture solutions, 3 ~ 4 parts
Turf mixes activated sludge, molasses, water, high-temperature sterilization, culture solution is added in, in 25 ~ 30 DEG C of temperature, 160 ~ 180r/
Ferment 1 ~ 2d under the conditions of min, adds turf, after mixing, dry at 40 ~ 50 DEG C of temperature, and crushes, and must crush
Object, in mass ratio 30 ~ 40：5 ~ 6, by step（2）Spare mixed liquor, crushed material are mixed, and with 200 ~ 300r/min into
10 ~ 15min is to get soil sterilants for row stirring.
Example 1
Phosphate buffer is in mass ratio 1：2：5, sodium dihydrogen phosphate, disodium hydrogen phosphate, water are mixed to get phosphoric acid buffer
Liquid.
Solid medium is to count in parts by weight, take 40 parts of water, 10 parts of agar, 10 portions of mannitol, 5 parts of calcium carbonate, 0.2 part
Potassium dihydrogen phosphate, 0.2 part of magnesium sulfate, 0.2 part of sodium chloride, 0.1 part of calcium sulfate are mixed, and high-temperature sterilization is to get solid culture
Base.
Beef extract-peptone tablet is to count in parts by weight, takes 50 parts of water, 15 parts of agar, 10 parts of peptones, 5 parts of chlorinations
Sodium, 3 portions of beef extracts are mixed, and high-temperature sterilization is to get beef extract-peptone tablet.
Fluid nutrient medium is to count in parts by weight, takes 40 parts of water, 10 portions of mannitol, 5 parts of calcium carbonate, 0.2 part of biphosphate
Potassium, 0.2 part of magnesium sulfate, 0.2 part of sodium chloride, 0.1 part of calcium sulfate are mixed, and high-temperature sterilization is to get fluid nutrient medium.
A kind of preparation method of soil sterilants, the preparation method include the following steps：
（1）In mass ratio 1：5, fly is put into the water that temperature is 0 DEG C and is ground, adds in the benzyl sulphonyl of water quality 0.01%
Fluorine grinds 25min, obtains lapping liquid, in mass ratio 1：4, lapping liquid, phosphate buffer are mixed, stood at 0 DEG C of temperature
25min, and carry out centrifugation 25min with 1000r/min, obtains supernatant, supernatant is heated, and temperature is to 90 DEG C, the time
3min, then centrifugation 25min is carried out with 1000r/min, supernatant is taken, supernatant is moved into carboxymethyl cellulose chromatographic column carries out sun
Ion-exchange chromatography, then eluted with phosphate buffer, sodium chloride solution, collect eluent；
（2）It counts in parts by weight, 3 parts of rhizoma atractylodis, 3 parts of grass-leaved sweetflags, 1 portion of folium artemisiae argyi is taken to be mixed, dries, places into powder in pulverizer
It is broken, 70 mesh sieve is crossed, obtains sieving, in mass ratio 1：3, sodium alginate, water are mixed, and is stirred with 200r/min,
The sieving of sodium alginate quality 10% is added, is stirred with 150r/min, obtains stirring mixture, stirring mixture is existed
It dries, crushes under temperature 50 C, cross 120 mesh sieve, obtain sieving a, in mass ratio 10：1, eluent, sieving a are mixed
It closes, and 20min is stirred with 200r/min, obtain mixed liquor, it is spare；
（3）In mass ratio 1：10, soil sample, sterile water in flower nursery are mixed, shaken cultivation 25min, quiet at 30 DEG C of temperature
1h is put, takes supernatant, 50% water dilution is added in into supernatant, in temperature 70 C water-bath 10min, dilution is obtained, by dilution
It is seeded in solid medium with oese, 1d, the bacterium colony of picking solid medium is cultivated, and connect in 20 DEG C of insulating boxs of temperature
Kind is isolated and purified to the flat lining out of beef extract-peptone, selects the bacterium colony of bacterium diameter maximum, in mass ratio 1：5, by bacterium diameter maximum
Bacterium colony, fluid nutrient medium mixed, in 25 DEG C of temperature, 120r/min shaking table culture 15h obtain culture solution；
（4）It counts in parts by weight, takes 40 parts of water, 40 parts of activated sludge, 10 parts of molasses, 5 parts of culture solutions, 3 parts of turfs, it will be active dirty
Mud, molasses, water are mixed, high-temperature sterilization, add in culture solution, in 25 DEG C of temperature, ferment 1d under the conditions of 160r/min, adds
Turf, it is after mixing, dry at 40 DEG C of temperature, and crush, obtain crushed material, in mass ratio 30：5, by step（2）It is standby
Mixed liquor, crushed material are mixed, and are stirred 10min with 200r/min to get soil sterilants.
Example 2
Phosphate buffer is in mass ratio 1：2：6, sodium dihydrogen phosphate, disodium hydrogen phosphate, water are mixed to get phosphoric acid buffer
Liquid.
Solid medium is to count in parts by weight, take 50 parts of water, 15 parts of agar, 15 portions of mannitol, 7 parts of calcium carbonate, 0.4 part
Potassium dihydrogen phosphate, 0.4 part of magnesium sulfate, 0.4 part of sodium chloride, 0.2 part of calcium sulfate are mixed, and high-temperature sterilization is to get solid culture
Base.
Beef extract-peptone tablet is to count in parts by weight, takes 60 parts of water, 25 parts of agar, 15 parts of peptones, 7 parts of chlorinations
Sodium, 4 portions of beef extracts are mixed, and high-temperature sterilization is to get beef extract-peptone tablet.
Fluid nutrient medium is to count in parts by weight, takes 50 parts of water, 15 portions of mannitol, 7 parts of calcium carbonate, 0.4 part of biphosphate
Potassium, 0.4 part of magnesium sulfate, 0.4 part of sodium chloride, 0.2 part of calcium sulfate are mixed, and high-temperature sterilization is to get fluid nutrient medium.
A kind of preparation method of soil sterilants, the preparation method include the following steps：
（1）In mass ratio 1：6, fly is put into the water that temperature is 4 DEG C and is ground, adds in the benzyl sulphonyl of water quality 0.02%
Fluorine grinds 30min, obtains lapping liquid, in mass ratio 1：5, lapping liquid, phosphate buffer are mixed, stood at 4 DEG C of temperature
30min, and carry out centrifugation 30min with 2000r/min, obtains supernatant, supernatant is heated, and temperature is to 98 DEG C, the time
4min, then centrifugation 30min is carried out with 2000r/min, supernatant is taken, supernatant is moved into carboxymethyl cellulose chromatographic column carries out sun
Ion-exchange chromatography, then eluted with phosphate buffer, sodium chloride solution, collect eluent；
（2）It counts in parts by weight, 4 parts of rhizoma atractylodis, 4 parts of grass-leaved sweetflags, 2 portions of folium artemisiae argyis is taken to be mixed, dries, places into powder in pulverizer
It is broken, 80 mesh sieve is crossed, obtains sieving, in mass ratio 1：4, sodium alginate, water are mixed, and is stirred with 300r/min,
The sieving of sodium alginate quality 15% is added, is stirred with 200r/min, obtains stirring mixture, stirring mixture is existed
It dries, crushes under temperature 60 C, cross 140 mesh sieve, obtain sieving a, in mass ratio 15：2, eluent, sieving a are mixed
It closes, and 30min is stirred with 300r/min, obtain mixed liquor, it is spare；
（3）In mass ratio 1：15, soil sample, sterile water in flower nursery are mixed, 30 min of shaken cultivation, quiet at 40 DEG C of temperature
2h is put, takes supernatant, 60% water dilution is added in into supernatant, in 80 DEG C of water-bath 15min of temperature, dilution is obtained, by dilution
It is seeded in solid medium with oese, 2d, the bacterium colony of picking solid medium is cultivated, and connect in 30 DEG C of insulating boxs of temperature
Kind is isolated and purified to the flat lining out of beef extract-peptone, selects the bacterium colony of bacterium diameter maximum, in mass ratio 1：6, by bacterium diameter maximum
Bacterium colony, fluid nutrient medium mixed, in 30 DEG C of temperature, 160r/min shaking table culture 18h obtain culture solution；
（4）It counts in parts by weight, takes 50 parts of water, 50 parts of activated sludge, 15 parts of molasses, 6 parts of culture solutions, 4 parts of turfs, it will be active dirty
Mud, molasses, water are mixed, high-temperature sterilization, add in culture solution, in 30 DEG C of temperature, ferment 2d under the conditions of 180r/min, adds
Turf, it is after mixing, dry under temperature 50 C, and crush, obtain crushed material, in mass ratio 40：6, by step（2）It is standby
Mixed liquor, crushed material are mixed, and are stirred 15min with 300r/min to get soil sterilants.
Example 3
Phosphate buffer is in mass ratio 1：2：5.5, sodium dihydrogen phosphate, disodium hydrogen phosphate, water are mixed and delayed to get phosphoric acid
Fliud flushing.
Solid medium is to count in parts by weight, take 45 parts of water, 12.5 parts of agar, 12.5 portions of mannitol, 6 parts of calcium carbonate,
0.3 part of potassium dihydrogen phosphate, 0.3 part of magnesium sulfate, 0.3 part of sodium chloride, 0.15 part of calcium sulfate are mixed, and high-temperature sterilization is to get solid
Body culture medium.
Beef extract-peptone tablet is to count in parts by weight, takes 55 parts of water, 20 parts of agar, 12.5 parts of peptones, 6 parts of chlorinations
Sodium, 3.5 portions of beef extracts are mixed, and high-temperature sterilization is to get beef extract-peptone tablet.
Fluid nutrient medium is to count in parts by weight, takes 45 parts of water, 12.5 portions of mannitol, 6 parts of calcium carbonate, 0.3 part of di(2-ethylhexyl)phosphate
Hydrogen potassium, 0.3 part of magnesium sulfate, 0.3 part of sodium chloride, 0.15 part of calcium sulfate are mixed, and high-temperature sterilization is to get fluid nutrient medium.
A kind of preparation method of soil sterilants, the preparation method include the following steps：
（1）In mass ratio 1：5.5, fly is put into the water that temperature is 2 DEG C and is ground, adds in the benzyl sulphur of water quality 0.015%
Acyl fluorides grinds 27.5min, obtains lapping liquid, in mass ratio 1：4.5, lapping liquid, phosphate buffer are mixed, in 2 DEG C of temperature
Lower standing 27.5min, and centrifugation 27.5min is carried out with 1500r/min, supernatant is obtained, supernatant is heated, temperature to 94
DEG C, time 3.5min, then centrifugation 27.5min is carried out with 1500r/min, supernatant is taken, supernatant is moved into carboxymethyl cellulose
Chromatographic column carries out cation-exchange chromatography, then is eluted with phosphate buffer, sodium chloride solution, collects eluent；
（2）It counts in parts by weight, 3.5 parts of rhizoma atractylodis, 3.5 parts of grass-leaved sweetflags, 1.5 portions of folium artemisiae argyis is taken to be mixed, dries, places into crushing
It is crushed in machine, crosses 75 mesh sieve, obtain sieving, in mass ratio 1：3.5, sodium alginate, water are mixed, and with 250r/min into
Row stirring, is added the sieving of sodium alginate quality 12.5%, is stirred with 175r/min, obtain stirring mixture, will stirred
Mixture is dried at 55 DEG C of temperature, is crushed, and is crossed 130 mesh sieve, is obtained sieving a, in mass ratio 12.5：1.5, by eluent, mistake
Sieve object a is mixed, and be stirred 25min with 250r/min, obtains mixed liquor, spare；
（3）In mass ratio 1：12.5, soil sample, sterile water in flower nursery are mixed, the shaken cultivation at 35 DEG C of temperature
27.5min stands 1.5h, takes supernatant, and 55% water dilution is added in into supernatant, in 75 DEG C of water-bath 12.5min of temperature, is obtained
Dilution is seeded to oese in solid medium by dilution, and 1.5d, picking solid are cultivated in 25 DEG C of insulating boxs of temperature
The bacterium colony of culture medium, and be seeded to the flat lining out of beef extract-peptone and isolate and purify, the bacterium colony of bacterium diameter maximum is selected, by quality
Than 1：5.5, the bacterium colony of bacterium diameter maximum, fluid nutrient medium are mixed, in 27 DEG C of temperature, 140r/min shaking table culture 16.5h,
Obtain culture solution；
（4）It counts in parts by weight, takes 45 parts of water, 45 parts of activated sludge, 12.5 parts of molasses, 5.5 parts of culture solutions, 3.5 parts of turfs, it will
Activated sludge, molasses, water are mixed, high-temperature sterilization, are added in culture solution, in 27 DEG C of temperature, are fermented under the conditions of 170r/min
1.5d adds turf, after mixing, dry under temperature 45 C, and crushes, and obtains crushed material, in mass ratio 35：
5.5, by step（2）Spare mixed liquor, crushed material are mixed, and are stirred 12.5min with 250r/min to get soil
Disinfectant.
Reference examples：The soil sterilants of company of Hefei City production.
Four mu of soil that environment is identical, long-term cropping is identical of selection are tested, and wherein first three mu is pressed before plant is colonized
100 parts/mu carry out spraying the soil sterilants that embodiment 1, embodiment 2, embodiment 3 obtain respectively, spray and plough deeply soil later；
4th mu sprays chemical agent as reference examples and carries out disinfection to soil, to above-mentioned four mu of soil grab sample after 1 ~ 2 week, and
It is observed in soil pest and disease damage detector, as a result such as table 1.
Table 1：
| Test event | Example 1 | Example 2 | Example 3 | Reference examples |
| Disinfection Effect | 97.9% | 98.4% | 97.4% | 84.3~87.2% |
| Safety | It is nontoxic to people and animals, plant | It is nontoxic to people and animals, plant | It is nontoxic to people and animals, plant | The nerve of people is stimulated, skin, liver to people have coup injury, corrode ozone layer, destroy ecological environment. |
| Residence time | Noresidue | Noresidue | Noresidue | 3 ~ 6 days |
By the data of example 1 ~ 3 it is found that example 2 is optimal example, the soil sterilants effect of gained is best.Summary,
The soil sterilants of the present invention is worth promoting.
Claims (9)
1. a kind of preparation method of soil sterilants, which is characterized in that the preparation method includes the following steps：
（1）Fly is put into the water that temperature is 0 ~ 4 DEG C and is ground, the benzyl sulfuryl fluoride of water quality 0.01 ~ 0.02% is added in, grinds
25 ~ 30min is ground, lapping liquid is obtained, lapping liquid, phosphate buffer is mixed, 25 ~ 30min is stood at 0 ~ 4 DEG C of temperature, and
Carry out 25 ~ 30min of centrifugation, obtain supernatant, supernatant is heated, temperature to 90 ~ 98 DEG C, 3 ~ 4min of time, then carry out from
25 ~ 30min of the heart, takes supernatant, and supernatant is moved into carboxymethyl cellulose chromatographic column carries out cation-exchange chromatography, then use phosphoric acid
Buffer solution, sodium chloride solution are eluted, and collect eluent；
（2）It counts in parts by weight, 3 ~ 4 parts of kaffir lily leaves, 3 ~ 4 parts of sansevieria trifasciata prain leaves, 1 ~ 2 part of grass-leaved sweetflag is taken to be mixed, is dried, then
It is put into pulverizer and crushes, be sieved, obtain sieving, sodium alginate, water are mixed, and is stirred, add alginic acid
The sieving of sodium quality 10 ~ 15%, is stirred, and obtains stirring mixture, stirring mixture is dried at 50 ~ 60 DEG C of temperature, powder
Broken, sieving obtains sieving a, mixes, and be stirred 20 ~ 30min eluent, sieving a, obtain mixed liquor, spare；
（3）Soil sample, sterile water in flower nursery are mixed, 25 ~ 30 min of shaken cultivation at 30 ~ 40 DEG C of temperature stands 1 ~ 2h,
Supernatant is taken, 50 ~ 60% water dilution is added in into supernatant, in 70 ~ 80 DEG C of 10 ~ 15min of water-bath of temperature, obtains dilution, it will be dilute
It releases liquid to be seeded in solid medium with oese, 1 ~ 2d is cultivated in 20 ~ 30 DEG C of insulating boxs of temperature, picking solid medium
Bacterium colony, and be seeded to the flat lining out of beef extract-peptone and isolate and purify, the bacterium colony of bacterium diameter maximum is selected, by the bacterium of bacterium diameter maximum
Fall, fluid nutrient medium is mixed, in 25 ~ 30 DEG C of temperature, 120 ~ 160r/min, 15 ~ 18h of shaking table culture obtain culture solution；
Activated sludge, molasses, water are mixed, high-temperature sterilization, culture solution is added in, in 25 ~ 30 DEG C of temperature, 160 ~ 180r/min
Under the conditions of ferment 1 ~ 2d, add turf, it is after mixing, dry at 40 ~ 50 DEG C of temperature, and crush, obtain crushed material,
By step（2）Spare mixed liquor, crushed material are mixed, and are stirred 10 ~ 15min to get soil sterilants.
2. the preparation method of soil sterilants according to claim 1, which is characterized in that the step（1）Middle fly, temperature
Mass ratio 1 for 0 ~ 4 DEG C of water：5 ~ 6, lapping liquid, phosphate buffer mass ratio 1：4~5.
3. the preparation method of soil sterilants according to claim 1, which is characterized in that the step（1）Middle phosphoric acid buffer
Liquid is in mass ratio 1：2：5 ~ 6, sodium dihydrogen phosphate, disodium hydrogen phosphate, water are mixed to get phosphate buffer.
4. the preparation method of soil sterilants according to claim 1, which is characterized in that the step（2）Middle chitosan, temperature
Spend the mass ratio 1 for 80 ~ 90 DEG C of water：3 ~ 4, eluent, sieving a mass ratio 10 ~ 15：1~2.
5. the preparation method of soil sterilants according to claim 1, which is characterized in that the step（3）It is native in middle flower nursery
The mass ratio 1 of sample, sterile water：10 ~ 15, the bacterium colony of bacterium diameter maximum, the mass ratio 1 of fluid nutrient medium：5~6.
6. the preparation method of soil sterilants according to claim 1, which is characterized in that the step（3）Middle solid culture
Base is to count in parts by weight, take 40 ~ 50 parts of water, 10 ~ 15 parts of agar, 10 ~ 15 portions of mannitol, 5 ~ 7 parts of calcium carbonate, 0.2 ~ 0.4 part
Potassium dihydrogen phosphate, 0.2 ~ 0.4 part of magnesium sulfate, 0.2 ~ 0.4 part of sodium chloride, 0.1 ~ 0.2 part of calcium sulfate are mixed, high-temperature sterilization,
Up to solid medium.
7. the preparation method of soil sterilants according to claim 1, which is characterized in that the step（3）Middle beef extract egg
White peptone tablet is to count in parts by weight, takes 50 ~ 60 parts of water, 15 ~ 25 parts of agar, 10 ~ 15 parts of peptones, 5 ~ 7 parts of sodium chloride, 3 ~ 4
Part beef extract is mixed, and high-temperature sterilization is to get beef extract-peptone tablet.
8. the preparation method of soil sterilants according to claim 1, which is characterized in that the step（3）Middle Liquid Culture
Base is to count in parts by weight, take 40 ~ 50 parts of water, 10 ~ 15 portions of mannitol, 5 ~ 7 parts of calcium carbonate, 0.2 ~ 0.4 part of potassium dihydrogen phosphate,
0.2 ~ 0.4 part of magnesium sulfate, 0.2 ~ 0.4 part of sodium chloride, 0.1 ~ 0.2 part of calcium sulfate are mixed, and high-temperature sterilization is to get Liquid Culture
Base.
9. the preparation method of soil sterilants according to claim 1, which is characterized in that the step（4）In by weight
Number meter, takes 40 ~ 50 parts of water, 40 ~ 50 parts of activated sludge, 10 ~ 15 parts of molasses, 5 ~ 6 parts of culture solutions, 3 ~ 4 parts of turfs, step（2）It is standby
The mass ratio 30 ~ 40 of mixed liquor, crushed material：5~6.
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| CN107216191A (en) * | 2017-05-25 | 2017-09-29 | 兰溪市普润斯水产养殖技术有限公司 | A kind of flowers chemical fertilizer and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1312292A (en) * | 2001-02-23 | 2001-09-12 | 辽宁天成生物制药研究所 | Preparation process of gene antibiotic peptide medicine from transgenic antibiotic peptide fly maggot |
| CN102485882A (en) * | 2010-12-02 | 2012-06-06 | 河南省农业科学院 | Method for cultivating predominant bacteria of fungus in alkaline soil |
| CN106148225A (en) * | 2016-06-29 | 2016-11-23 | 四川农业大学 | A kind of bacillus subtilis, its screening technique and purposes and biological preservative, its preparation method and fruit and vegetables corrosion protection method |
| CN107216191A (en) * | 2017-05-25 | 2017-09-29 | 兰溪市普润斯水产养殖技术有限公司 | A kind of flowers chemical fertilizer and preparation method thereof |
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