CN105439725A - Paenibacillus polymyxa pesticide-fertilizer for farm onsite fermentation and applications thereof - Google Patents
Paenibacillus polymyxa pesticide-fertilizer for farm onsite fermentation and applications thereof Download PDFInfo
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- CN105439725A CN105439725A CN201510951101.1A CN201510951101A CN105439725A CN 105439725 A CN105439725 A CN 105439725A CN 201510951101 A CN201510951101 A CN 201510951101A CN 105439725 A CN105439725 A CN 105439725A
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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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- C05B7/00—Fertilisers based essentially on alkali or ammonium orthophosphates
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Abstract
The invention relates to a paenibacillus polymyxa pesticide-fertilizer for farm onsite fermentation, a preparation method, and applications thereof. The pesticide-fertilizer comprises the following raw materials: paenibacillus polymyxa powder, colistin sulfate, natamycin, L-alanine, valine, yeast powder, corn steep liquor powder, disodium hydrogen phosphate, sodium chloride, fructose, and brown sugar. The colistin sulfate and natamycin in the insecticide-fertilizer can inhibit the growth of fungi (such as pathogenic fungi, yeast, etc.) and gram negative bacteria (such as colibacillus, salmonella, etc.) during the fermentation process. The spore germinator (alanine, fructose, and valine) can rapidly promote the germination of spores of paenibacillus polymyxa and improve the germination rate of spores. The provided pesticide-fertilizer can be applied to the onsite fermentation and propagation in a simple and crude farm. The required equipment is simple. The using amount of bacterium is little. The cost is low. The fermentation broth obtained through onsite fermentation has the advantages of high content and good activity. The insecticide-fertilizer can prevent disease, promote growth, and increase output and improve product quality, and can be used as a microbial pesticide, and the effect is stable.
Description
Technical field
The present invention relates to microbial fertilizer and microbial pesticide field, particularly relate to a kind of the Paenibacillus polymyxa medicine fertilizer and the application thereof that are applied to the fermentation of scene, farm.
Background technology
Along with the development of existing agricultural policy; farm size; the trend maximized is strengthened further, and bacillus is current biological pesticide, a kind of microorganism of most study in bio-feritlizer; it has broad spectrum antibiotic activity and extremely strong anti-adversity ability; its vegetative cell can be grown surely in the procreation of fungal pathogen mycelium surface, effectively suppresses pathogen growth, therefore; occur in control disease, promote plant strain growth, improve the aspects such as crop yield and show wide application prospect.
Many sticky class bud pole bacterium are a kind of sporiferous gram positive bacteriums, belong to class bud and to satisfy Bacillaceae.Its bacterium colony is many in pale yellow or white, protuberance, and thick, surface wettability is smooth, and thalline is shaft-like, and G ten C content is 40% one 50%; Utilize peritrichous to move, produce ellipse bud and embrace expanding to embrace in ascus; The most suitable growth pH is 7.0, and optimum temperuture is 28-35 DEG C; Amphimicrobian, decomposition glucose and other carbohydrates produce acid, and aerogenesis sometimes, without soluble pigment on nutrient agar medium.
The bacterial strain of many sticky class Bacillus has bud and to satisfy some general character of Bacillus genus strain, not only there is significant Biocontrol Potential, the bud that can also produce heat-resistant inverse is embraced, be conducive to the production of biocontrol fungicide, formulation and survive in the environment, surely grow and breeding, the important component part (parthaPatra of Plant diseases Biocontrol microorganism, etal, 2006).Many sticky class buds bacillus that satisfies has non-virulent to plant, has prophylaxis effect and growth-promoting functions simultaneously, is widely used in agriculture production as biocontrol microorganisms.Many sticky class bud pole bacterium can produce the multiple antimicrobial substance such as peptide class, protein-based, ucleosides, Pyrazine and phenols, can prevent and treat plurality of plant diseases (Lee another name for Sichuan Province etc., 2007; Yu Shuchi etc., 2004).
In addition, many sticky class bud pole bacterium have the ability (Xiao Chunqiao etc. moving from soil to plant root and surely grow, 2004), and act as host plant by fixed nitrogen and molten phosphorus nutritive ingredient (Chen Zhiyi etc. are provided, 2001), promote plant strain growth, improve the effect of plant disease-resistant pathogenic microorganism, many sticky class bud pole bacterium sphere of action are wide, pathogenetic bacteria can be prevented and treated, pathogenic fungi, root knot nematode (Xiao Chunqiao etc., 2004) the multiple harmful microorganism such as, mechanism of action is various, existing antimicrobial substance works, there is again the direct effect of microorganism, induction host plant is likely made to produce disease resistance (Zhao Shuan etc., 2008).Many sticky class buds satisfy bacillus as biocontrol microorganisms to be applied to a lot of field, and the secondary metabolite that this bacterium produces can suppress a lot of phytopathogen, and can promote plant growth, and induction crop produces systemic disease resistance.How sticky class bud pole bacterium w3 and Y2-11-1 that the screening such as Tong Yunhui obtains anti-Botrytis cinerea has extremely significant system induction effect to tomato plant botrytis resistant, it can be grown well surely at tomato body surface, surely the time of growing reaches 10-15d (Tong Yunhui etc., 2004).The research such as Zhang Zhen find many sticky class buds satisfy bacillus A2 to cucumber at seedling stage samping off and vegetative period blight have good prevention effect, and this bacterium can promote the growth of cucumber, increases dry matter content.The how sticky class bud of 0.1 hundred million cfu/g bacillus granula subtilis (health ground flower bud obtain) of satisfying is the microbial bactericide researched and developed by East China University of Science, the machine-processed Suppressing phytopathogens such as disease resistance and Promoting plant growth (Wei Honggang etc., 2008) is produced by antimicrobial substance, rhizosphere site competition, inducing plant.Many sticky class bud pole bacterium have protection effect good, to the advantage such as person poultry safety and non-environmental-pollution, be classified as a level security bacterial classification by the Ministry of Agriculture in China.After beet is inoculated endogenetic polymexa sporeformer S-7, it significantly improves beet leaf Net Photosynthetic Rate, makes beet leaf rate of photosynthisis maintain a higher level, thus adds the sugar content (Shi Yingwu etc., 2009) of beet.
The secondary metabolite kind that many sticky class bud pole bacterium produce is more, complicated component.Zhang Daojing etc. are separated to 8 kinds of compounds in the ethanol extraction of many sticky class bud pole fermented liquids, be respectively uridylic, thymus pyrimidine, Sophoricol, 5,4.-dihydroxyl-7-methoxy flavone, 2,4-DI-tert-butylphenol compounds, p-Hydroxybenzaldehyde, palm fibre put acid, meso-2,3-butyleneglycol (Zhang Daojing etc., 2008).Wang Zhiwen etc. utilize macroporous adsorbent resin to satisfy to many sticky classes the Adsorption and desorption performance of antifungus active substance that bacillus Cp-S316 produces, and determine the some properties (Wang Zhiwen etc., 2007) of antifungus active substance.The antimicrobial spectrum of Paenibacillus polymyxa is comparatively wide, and meta-bolites is abundanter, can produce multiple antibacterial substance.Paenibacillus polymyxa has the ability suppressing Plant diseases, can be made into biotechnological formulation disease control after plant flowers portion, leaf, limb, root and fruits and vegetables Bian; Use the fermented liquid of Paenibacillus polymyxa, farm crop are sprayed, dress seed, fill with root etc., plant is all produced to effect of growth promoting effects.In field of biological control, Paenibacillus polymyxa application is a lot.Although many with the microbial pesticide product that genus bacillus is main preparation, but all things considered, also exist comprise research and development with produce disconnect, formulation difficulty, constant product quality is poor, production technique is backward, homogeneity of product is not high, manufacturing enterprise's small scale, the problem that application difficulty is large, except a little, based on wettable powder, although this formulation is convenient to store and transport, in production application, there is a lot of restrictive factor in current genus bacillus.As most using gemma as the unitary basis weighing effective constituent in product in the wettable powder used, if gemma can not be sprouted in time, application effect is then not good, as Wang Qian utilize subtilis TR21 to prevent and treat Brazilian any of several broadleaf plants blight time, pre-stage test utilizes TR21 bacterial strain NB fermentation liquid irrigating root and axil inoculation effectively can prevent and treat blight, but utilize gemma to prepare the filling root of wettable powder and axil effect undesirable, showing the pure gemma after removing fermented liquid may because sprouts difficulty and lack effective constituent in fermented liquid and make preventive effect decline.In addition, in production practice, along with the growth of crop, need regularly constantly to supplement biocontrol microorganisms and could obtain good preventive effect, but existing wettable powder price is general higher, and the dissolving of wettable powder needs to use a large amount of water, and spreading along with Plant diseases, source of clean water also becomes the factor that restriction biological prevention and control agent uses, the pathogenic bacteria that can not directly kill in water is dissolved in water unlike gemma with chemical agent, if thus use and used biocontrol microorganisms by the water body of pathogen contamination, also spreading of disease can be promoted.
In order to can production be beneficial to, transport, the sporeformer that we produce in product mainly exists with the state of spore, but it is applied to crop, soil, in water body, the mainly vegetative state cell all needed, as used bacillus preparation on aquatic products, before use can through overactivation, even if the genus bacillus of gemma state is converted into vegetative state cell, but activation traditionally mainly adds some brown sugar, or glucose, the nutritive substance of substratum and so on induces its growth and breeding, as bacillus preparation: brown sugar: water carries out " activation " with the ratio of 1:1:10, but there is the problem of the following aspects in " activation " traditionally:
One, be the yeast easily made after high nutritional medium adds water in air and water, milk-acid bacteria breeding rapidly, consumes substratum Middle nutrition, reduces pH in nutrient solution rapidly, suppresses the Germination and growth of gemma;
Two, be that gemma sprouting needs the time long in early stage, general needs 4 hours, gemma just can start to sprout, and now, because the amount reproduction of yeast and milk-acid bacteria causes pH to decline rapidly in " activation " liquid, can suppress sprouting and the breeding of gemma, thus greatly affect gemma sprouting;
Three, be also easy infection intestinal bacteria in high nutritional medium, harmful miscellaneous bacteria such as the plant disease fungus of original work, causes using rear weak effect and has still caused retroaction;
Four, be that sporeformer consumption is large, effect is unstable.
Along with soil allows circulation; country advocates farm formula and manages; farm size; the trend maximized will be strengthened further; numerous fermentation biocontrol microorganisms then suitable demand applying agricultural, commodities market is carried out expanding in scene, farm, but is not up to the present also disclosed and is relevantly a kind ofly applied to the fermentation of scene, farm and expands numerous Paenibacillus polymyxa medicine fertilizer that better can control miscellaneous bacteria and promotion gemma fast-germination and the correlation technique applied thereof.
Summary of the invention
In order to overcome the shortcoming of prior art, the invention provides and be a kind ofly applied to the fermentation of scene, farm and expand and numerous better can control miscellaneous bacteria and promote that the Paenibacillus polymyxa medicine of gemma fast-germination is fertile and apply, be to provide specifically one can be applicable to farm scene fermentation expand numerous, tennecetin and polymyxin is utilized to suppress pathogenic fungi, the fungies such as yeast and intestinal bacteria, the growth of the Gram-negative bacterias such as Salmonellas, utilize L-Ala, fructose, the gemma germinants such as α-amino-isovaleric acid, the sprouting of gemma can be promoted fast, improve gemma germination efficiency, reduce gemma sprout time, thus use Paenibacillus polymyxa breeding fast and occupy dominant bacteria, and better suppress other miscellaneous bacteria and harmful bacteria, therefore, a kind of bacteria using amount can be obtained few, cost is low, the fermented liquid bacterial content of on-the-spot fermentation is high, active high, miscellaneous bacteria is few, there is disease prevention growth-promoting, increasing yield and improving quality, effect stability, can be used as the Paenibacillus polymyxa fermented liquid of microbial pesticide and Fertilizer application.
To achieve these goals, the technical solution adopted in the present invention is: a kind of Paenibacillus polymyxa medicine for the fermentation of scene, farm is fertile, and the raw material preparing this Paenibacillus polymyxa medicine fertilizer is:
Paenibacillus polymyxa bacterium powder 50-200 part, colistine sulfate 10-50 part, tennecetin 1-2 part, ALANINE 3-40 part, α-amino-isovaleric acid 8-50 part, yeast leaching powder 50-200 part, peptone 50-200 part, Sodium phosphate dibasic 1-10 part, sodium-chlor 1-10 part, glucose 20-200 part, fructose 20-50 part, brown sugar 20-200 part.
In order to achieve the above object, the preparation method for the solution shallow lake genus bacillus medicine fertilizer of scene, farm fermentation comprises following steps:
First by colistine sulfate 10-50 part, tennecetin 1-2 part, ALANINE 3-40 part, α-amino-isovaleric acid 8-50 part, yeast leaching powder 50-200 part, peptone 50-200 part, Sodium phosphate dibasic 1-10 part, sodium-chlor 1-10 part, glucose 20-200 part, fructose 20-50 part, brown sugar 20-200 part, place in sealing stirred pot and stir 20-30 minute, mix, then after Paenibacillus polymyxa bacterium powder 50-200 part being added and stirring in stirred pot, packing 500 grams is often wrapped, vacuumize packaging, the Paenibacillus polymyxa medicine namely obtained for scene fermentation is fertile;
Wherein, the application of described a kind of Paenibacillus polymyxa medicine fertilizer for the fermentation of scene, farm, comprises the following steps:
the fertile method of carrying out fermenting at scene, farm of this Paenibacillus polymyxa medicine is: the water-wet first the Paenibacillus polymyxa medicine of above-mentioned preparation fertilizer being added its 3-5 times weight is mixed well, water is supplied again after 30 minutes, to reach the ratio that Paenibacillus polymyxa medicine ratio that is fertile and water is 1:10 to 1:30, after adjusting PH to 8-8.2, start to cultivate, within first 12 hours, per minute air flow liquid-gas ratio is 1:0.6, after 12 hours, air flow is 1:1, culture temperature is 20-35 DEG C, cultivate after 16-24 hour, the fermented liquid of the Paenibacillus polymyxa medicine fertilizer of on-the-spot fermentation can be obtained;
the application of Paenibacillus polymyxa medicine fertilizer for the fermentation of scene, farm be the fermented liquid of the Paenibacillus polymyxa medicine fertilizer of on-the-spot fermentation in arable farming to soil-borne disease, the application of leaf diseases;
the application of Paenibacillus polymyxa medicine fertilizer for the fermentation of scene, farm be the fermented liquid of the Paenibacillus polymyxa medicine fertilizer of on-the-spot fermentation in arable farming as base fertilizer, topdress and foliage fertilizer and the application of spreading manuer in holes and dressing seed;
Wherein, for soil-borne disease as tobacco bacterial wilt, balck shank, watermelon blight, cucumber fusarium axysporum etc., by step
the fermented liquid dilution 500-800 of preparation doubly after adopt the method for filling with root to prevent and treat, when transplanting and their early stage medication, each is 0.5-0.8 kg/acre by fermentation liquid measure;
Wherein, for leaf diseases as rice blast, rice sheath blight disease, powdery mildew of strawberry etc., by step
the fermented liquid dilution 200-500 of preparation doubly adopts the method for spraying to prevent and treat, and starts medication at the initial stage of a disease, uses once every 7 days, 3-5 time altogether;
Wherein, during as base fertilizer, by step
fermented liquid and other fertilizers of preparation and chemical fertilizer with the use of, be 5-20 kg/acre by fermentation liquid measure at every turn;
Wherein, as when topdressing, by step
the fermented liquid dilution 500-1000 of preparation doubly fills with root, and each is 0.1-0.3 kg/acre by fermentation liquid measure;
Wherein, as foliage fertilizer, by step
the fermented liquid dilution 300-800 times of foliage-spray of preparation, till positive and negative blade face all squirts, each is 0.3-0.5 kg/acre by fermentation liquid measure;
Wherein, during for dressing seed, by step
doubly, dress seed the fermented liquid dilution 200-500 of preparation 2-12 hour.
Wherein, the spore content of described Paenibacillus polymyxa bacterium powder is not less than 10,000,000,000 cfu/g.
Wherein, the water content of each component raw material of described a kind of gemma medicine fertilizer for the fermentation of scene, farm is lower than 5%.
Wherein, described Paenibacillus polymyxa bacterium powder can adopt prior art, such as wettable powder, common bacterium powder, moisture dispersion agent all can, the invention provides following methods:
Take out Paenibacillus polymyxa culture presevation pipe, draw flat board with LB solid medium and recover, cultivate 48 hours for 37 DEG C.Under flat board in the single colony inoculation of picking 50 milliliters of LB substratum, in incubator, 37 DEG C shake cultivation 24 hours.Seed adopts the inoculum size of 5%, be seeded in the large triangular flask of 5L, liquid amount is the LB substratum of 2L, 20-28 hour is cultivated in 37 DEG C of concussions, detect its bacterial concentration, viable bacteria content is more than 4,000,000,000/milliliter, namely can be used as bacillus liquid body strain inoculation to 1000L fermention medium, inoculum size is 5%, open and stir 120r/min, per minute air flow is liquid/gas 1:0.8, cultivate 16-24 hour for 37 DEG C, treat that spore content is not less than 8,000,000,000/milliliter, fermentation can be terminated, add diatomite spraying dry, water content is lower than 5%, bacteria concentration is regulated to be 10,000,000,000/gram, pulverize 100 orders and namely obtain genus bacillus bacterium powder.Wherein said LB substratum: peptone 5g, extractum carnis 3g, sodium-chlor 5g, water 1000mL, pH7.2, add agar 2% in solid medium; Wherein said genus bacillus liquid bacterium culture medium: Semen Maydis powder 10g/L, Zulkovsky starch 20g/L, corn starch 10g/L, bean cake powder 44g/L, magnesium sulfate 0.5g/L, manganous sulfate 0.5g/L, potassium primary phosphate 0.5g/L, dipotassium hydrogen phosphate 0.8g/L, calcium carbonate 2.5g/L, pH7.2.The condition of each medium sterilization is: 0.10-0.15MPa, 121 DEG C of sterilizings 30 minutes.
Containing tennecetin in this Paenibacillus polymyxa medicine fertilizer, tennecetin is 26 kinds of antibiotic one of polyene macrolide antifungal, produce primarily of 3 kinds of streptomycetes, just exert to add streptomycete (Streptomyceschattanovgensis), Streptomyces natalensis Streptomyxesnatalensis respectively) and brown yellow spore streptomycete (Streptomyxesgilvosporeus), through fermentation, extraction, the refining biological preservative with wide spectrum, economic benefits and social benefits obtained.The food spoilage that it both can suppress various mould, the growth of yeast causes, again can the generation of Antifungi toxin, prevents from causing toxin type to poison by food.1996, China's approval tennecetin was used for the surface anticorrosion of cheese, meat product, moon cake, cake, fruit juice primary pulp and easily mouldy food-processing vessel.Tennecetin also goes through directly to add in fermented wine, Yoghourt and salad dressing, limits the quantity as 10mg/kg.Tennecetin, to human non-toxic, and without mutagenesis, carcinogenic, teratogenesis and sensitization, and is difficult to be absorbed by mammalian digestive tract.To the tennecetin of detoxification high dosage, more than 90% discharges with ight soil with prototype and degradation production thereof.Research shows, under natural condition, microorganism is difficult to produce the resistance to tennecetin.The clinical medicine range of application of tennecetin is also very extensive, may be used for the fungi infestation of anti-skin, mucous membrane, vagina and lung.Tennecetin is the antimycotic biological preservative uniquely got the Green Light in the world, is a kind of efficient, safe New Biological Preservatives.Tennecetin is that a kind of polyene hydrocarbon macrolide is antifungal agents based, polyenoid is a plane macrolide ring texture, interacts have high affinity with sterol compound on cell, and trigger cell membrane structure changes and breaks thus, cause the seepage of entocyte, make necrocytosis.But there are not these sterol compounds, so tennecetin does not act on bacterium in some microorganism such as the cell walls of bacterium and cytoplasmic membrane.
Colistin, Polymyxin E is had another name called containing colistin (Colistin) in Paenibacillus polymyxa medicine fertilizer of the present invention.Nineteen fifty, from Fukushima county soil, be separated to by Japanese hill Kang Fu etc. a kind of lock ring shape peptide antibiotics produced in strain aerobacillus polymyxa Donker colistin mutant (Bacilluspolymyxavarcolistin) nutrient solution.It combines with the free phosphoric acid salt of lipoprotein in sensitive organism endochylema film, and endochylema film tension is reduced, and permeability increases, and causes endochylema film to lose barrier action and cell Nei Piao ridge, pyrimidine, Nucleotide etc. is leaked outside and dead, genus sterilization class microbiotic.Colistin has stronger anti-microbial effect to most of Gram-negative bacteria, as intestinal bacteria, Salmonellas, pseudomonas etc., but to gram-positive microorganism and fungi nearly unavailable.
Containing L-Ala in Paenibacillus polymyxa medicine fertilizer of the present invention, α-amino-isovaleric acid, the gemma germinants such as fructose, occurring in nature gemma sprouts the impact by exogenous nutrition material, this kind of nutritive substance is referred to as " germinant ", under normal circumstances, after the gemma germinant mixing that gemma and promotion are sprouted within the several seconds, germinant strides across the surface of gemma clothing and cortex arrival intine fast, interact with the receptor protein be positioned on intine, gemma is excited to enter germinating, then a succession of sprouting event is started, germination process afterwards can not stop because removing germinant.
Containing ALANINE in the present invention, ALANINE is widely used in foodstuff production as the efficient gemma germinant of one, and multiple field such as inspection and quarantine, is not used in biological pesticide, in the research field of bacterial manure.Add external source carbohydrate substance to improve bud and to satisfy germination efficiency (clementsandMoir1998; Paidhungatandsetlow, 2001).Meeannetal (1996) and Thanhetal (2005) points out that, when ALANINE exists, monose can promote that bud embraces sprouting.Equally, in the present invention, certain density fructose can promote that together with ALANINE bud is embraced efficiently and sprouts completely.
Containing a large amount of yeast leaching powder in the present invention, peptone, brown sugar, is all the nutrition agent being applicable to promoting fermentation of bacillus breeding, once gemma is sprouted, can promotes breeding and the multiplication of genus bacillus fast.
Paenibacillus polymyxa in Paenibacillus polymyxa medicine fertilizer in the present invention from Trace Elements in Rhizosphere Soil of Flue-Cured Tobacco, is separated the effect that has obtained prevent and treat Powdery Mildew, black shank, gray mold, rice blast, cucumber fusarium axysporum, early blight of tomato, pimento anthrax, watermelon sclerotium disease, wheat scab, watermelon blight etc. prevent and treat the genus bacillus of multiple diseases.
The present invention adopts the beneficial effect of such scheme: Paenibacillus polymyxa medicine is fertile first carries out wetting mixing in the ratio of 1:3-1:5 with water, after gemma germinant now can fully contact with gemma, thus impel it to sprout soon, through overtesting, after 30 minutes, gemma germination rate reaches more than 75%, moisturizing is to Paenibacillus polymyxa bacterial manure and water first in the ratio of 1:10-1:30 again, regulate pH to 8-8.2, ventilation starts to cultivate, under higher pH, expanding the growth of the good lactic acid bacteria inhibiting of numerous earlier fermentation energy, the tennecetin wherein contained in Paenibacillus polymyxa medicine fertilizer and polymyxin can suppress preferably in water with contained yeast in the air that passes into, pathogenic fungi and intestinal bacteria, the growth of the harmful bacterias such as Salmonellas, and the yeast powder in Paenibacillus polymyxa medicine fertilizer, peptone, brown sugar, it is all the nutrition agent being applicable to promoting fermentation of bacillus breeding, once gemma is sprouted, breeding and the multiplication of genus bacillus can be promoted fast.Thus increase the thalline content of its genus bacillus fast, thus make genus bacillus breed and occupy dominant bacteria fast, and better suppress other miscellaneous bacteria and harmful bacteria.
Therefore, the fertile water used that ferments at the scene of Paenibacillus polymyxa medicine of the present invention, air, container does not need the sterilizing that carries out disinfection, in the on-the-spot fermentation culture process in farm, miscellaneous bacteria can be suppressed preferably, reach the bacillus polymyxa fermented liquid of in-situ batch culture higher concentration.Paenibacillus polymyxa medicine for the fermentation of scene, farm of the present invention is fertile, and production technique is simple, can at simple and crude farm execute-in-place in on-the-spot fermenting process, facility demand is simple, simple to operate, easy to implement, can be mass and apply, bacteria using amount is few, and cost is low, the fermented liquid bacterial content of on-the-spot fermentation is high, and active high, miscellaneous bacteria is few, there is disease prevention growth-promoting, increasing yield and improving quality, can be used as microbial pesticide and Fertilizer application, effect stability.
Embodiment
Embodiment 1 illustrates the superiority of the gemma " activation " that the Paenibacillus polymyxa medicine of the present invention scene fermentation is fertile and traditional below by test
Test group: colistine sulfate 15 parts, tennecetin 1 part, ALANINE 10 parts, α-amino-isovaleric acid 18 parts, yeast leaching 100 parts, powder, peptone 100 parts, Sodium phosphate dibasic 2 parts, 2 parts, sodium-chlor, glucose 22 parts, 30 parts, fructose, 100 parts, brown sugar is placed in sealing stirred pot and is stirred 20-30 minute, mix, again 100 parts, Paenibacillus polymyxa bacterium powder is added after stirring in stirred pot, get 1kg to do experiment, the water-wet first Paenibacillus polymyxa medicine fertilizer being added 4kg is mixed well, water is supplied again to reach the ratio (altogether the water of 20kg) that Paenibacillus polymyxa medicine ratio that is fertile and water is 1:20 after 30 minutes, after adjusting PH to 8.2, start to cultivate, within first 12 hours, per minute air flow liquid-gas ratio is 1:0.6, after 12 hours, air flow is 1:0.8, culture temperature is 20-35 DEG C, cultivate the fermented liquid of the Paenibacillus polymyxa medicine fertilizer being on-the-spot fermentation after 24 hours,
Control group: yeast leaching 100 parts, powder, peptone 100 parts, Sodium phosphate dibasic 2 parts, 2 parts, sodium-chlor, glucose 22 parts, 100 parts, brown sugar, place in sealing stirred pot and stir 20-30 minute, mix, again 100 parts, bacillus megaterium bacterium powder is added after stirring in stirred pot, get 0.852kg to do experiment, first prepared by control group the fertile water-wet adding its 4kg of Paenibacillus polymyxa medicine to mix well, water is supplied again to reach the ratio (water of 20kg) that Paenibacillus polymyxa medicine ratio that is fertile and water is 1:20 after 30 minutes, after adjusting PH to 8.2, start to cultivate, within first 12 hours, per minute air flow liquid-gas ratio is 1:0.6, after 12 hours, air flow is 1:0.8, culture temperature is 20-35 DEG C, cultivate the fermented liquid of the Paenibacillus polymyxa medicine fertilizer being scene " activation " after 24 hours,
Under identical culture condition, detect the pH of fermented liquid in its 24 hours culturing process, gemma, total viable count, the changing conditions of miscellaneous bacteria number is as accompanying drawing 1, and accompanying drawing 2, shown in accompanying drawing 3.
Can find out that from accompanying drawing 1 the pH change of test group is slower, pH maintains more than 7 always, and the pH of control group is just reduced to less than 5 after 8 hours fast, after 24 hours, pH is reduced to less than 4, at this moment be comparatively applicable to yeast, the growth of milk-acid bacteria, lower pH can suppress sprouting and the growth of gemma.Can obviously find out from accompanying drawing 2, after 4 hours, its gemma germination rate reaches more than 90% to the gemma of test group, and the gemma of control group ferments 24 hours its gemma germination rates later just to about 50%.
From accompanying drawing 3, obviously can see that the fermentation of test group is after 24 hours, its total count about reaches 2,100,000,000 CFU/ml, substantially lower than 0.1 hundred million CFU/ml after assorted fungal counting, and total bacterial content of control group is about 1,100,000,000 CFU/ml, but miscellaneous bacteria content then reaches 3.8 hundred million CFU/ml.Therefore illustrate that test group adds sanitas: colistine sulfate, tennecetin, with gemma germinant L-Ala, α-amino-isovaleric acid, fructose etc. just significantly can suppress varied bacteria growing, and promote the Fast-propagation of row Paenibacillus polymyxa, its action effect is very significant.
Wherein, accompanying drawing 1 is test group and the change curve (the fertile on-the-spot pH change curve fermented of Fig. 1 Paenibacillus polymyxa medicine) contrasting the on-the-spot middle pH that spreads cultivation.
Wherein, accompanying drawing 2 is change curves (the fertile on-the-spot remaining gemma change curve that ferments of Fig. 2 Paenibacillus polymyxa medicine) of the remaining gemma of test group and control group.
Wherein, accompanying drawing 3 is change curves (change curve of Fig. 3 Paenibacillus polymyxa medicine fertile on-the-spot ferment total count and miscellaneous bacteria number) of test group and control group total viable count and miscellaneous bacteria number.
Embodiment 2
Utilize the fermented liquid of the Paenibacillus polymyxa medicine fertilizer of embodiment 1 test group situ production in flue-cured tobacco as the effect of medicine fertilizer
1. test site: Bai Ju village, beachhead town, Longhui County, Hunan Province Cotton Varieties by Small Farming Households ground
2. test method: flue-cured tobacco cultivars: middle cigarette 99-10-1, establishes 3 process altogether, repeats for 4 times.Test area 24 mu, peripheral hardware protection row.Test group: the conventional flue-cured tobacco Special basal fertilizer (N-P2O5-K2O=10-10-25) i.e. of fermented liquid+60% 24 kilograms of special fertilizer for tobacco of Paenibacillus polymyxa medicine fertilizer; The mu consumption of the fermented liquid of Paenibacillus polymyxa medicine fertilizer is 4 kilograms: during flue-cured tobacco transplanting seedling, with fermented liquid 800 times of liquid irrigating roots of Paenibacillus polymyxa medicine fertilizer with to be stained with root each once; Within after slow seedling 20 days, fill with second time with fermented liquid 1000 times of liquid of Paenibacillus polymyxa medicine fertilizer, spray third time and the 4th time every 20 days then continuously;
Control group 1: the conventional flue-cured tobacco Special basal fertilizer (N-P2O5-K2O=10-10-25) i.e. of activation solution+60% 24 kilograms of special fertilizer for tobacco of the bacillus polymyxa at control group scene; The mu consumption of the activation solution of the bacillus polymyxa of control group is 4 kilograms: during flue-cured tobacco transplanting seedling, with activation solution 800 times of liquid irrigating roots of the bacillus polymyxa of control group with to be stained with root each once; Within after slow seedling 20 days, fill with second time with activation solution 1000 times of liquid of the bacillus polymyxa of control group, spray third time and the 4th time every 20 days then continuously;
Control group 2 (CK): conventional flue-cured tobacco Special basal fertilizer, the consumption of flue-cured tobacco Special basal fertilizer is 40 kgs/acre, and fertilizing method is bottom application, fills with root with test group with clear water simultaneously;
3. test-results:
Because flue-cured tobacco is all planted in continuous 2 years in the soil in Bai Ju village, although also plant every year one season late rice, but its bacterial wilt and balck shank sickness rate higher and, after test group uses the fermented liquid of Paenibacillus polymyxa medicine fertilizer production, plant height, stem girth, pitch, leaves number, maximum leaf area is all greater than control group, the situation of the composite fertilizer of minimizing 40% is described as table 1, the fermented liquid of the Paenibacillus polymyxa medicine fertilizer of test group situ production effectively can improve the economical character of cigarette strain, promote cigarette strain robust growth, the effect of the activation solution of same Paenibacillus polymyxa is utilized then to have some improvement with blank group effect, but obviously and the effect of the fermented liquid of the Paenibacillus polymyxa medicine fertilizer of test group situ production have obvious gap.As shown in table 2 use the fermented liquid of the Paenibacillus polymyxa medicine fertilizer of test group cigarette strain bacterial wilt and balck shank sickness rate very low, and occurring degree significantly reduces, control group 1 and control group 2 then have obvious bacterial wilt and balck shank, also obviously there is disease after usage chain mycin and 50% fenaminosulf wettable powder;
Table 1 Paenibacillus polymyxa medicine fertilizer produce fermentation broth on tobacco plant height, stem girth, pitch, leaves number, maximum leaf area impact
Table 2 uses fermented liquid that Paenibacillus polymyxa medicine fertilizer produces to the prevention effect of land for growing field crops tobacco to balck shank and bacterial wilt
Fermentation broth on tobacco yield and output value the effect of increasing income of table 3 Paenibacillus polymyxa medicine fertilizer
| Process | Output (kg/acre) | The output value (unit/mu) | Average price (unit/kilogram) |
| Control group 2(CK) | 196.4 | 2718.2 | 13.84 7 --> |
| Control group 1 | 208.3 | 2926.6 | 14.05 |
| Test group | 266.7 | 4043.2 | 15.16 |
Use Paenibacillus polymyxa medicine fertilizer produce fermented liquid after tobacco premature ripening, improve quality, improve the output of tobacco leaf, average price, the output value, upper medium grade cigarette ratio to a great extent.Can analyze from table 3 and draw, est and pilot project yield of tobacco, the output value, average price be group more blank than control group 2(clear water respectively) exceed 60.4 kgs/acre, 1212.2 yuan/mu, 1.52 yuan/kilogram.Deduct the cost 440 yuan (wherein artificial 360 yuan) of increase, 772.2 yuan/mu can be had a net increase of.And namely use the activation solution of bacillus polymyxa relative to control group 1, then mu increases production 1116.6 yuan.And the control group 1 using the activation solution of bacillus polymyxa is more or less the same with the effect of clear water blank group 2, do not embody obvious volume increase advantage.
Embodiment 3
A kind of Paenibacillus polymyxa medicine for the fermentation of scene, farm is fertile, and the raw material preparing this Paenibacillus polymyxa medicine fertilizer is:
90 parts, Paenibacillus polymyxa bacterium powder, colistine sulfate 18 parts, tennecetin 1 part, ALANINE 12 parts, α-amino-isovaleric acid 22 parts, yeast leaching 100 parts, powder, peptone 100 parts, Sodium phosphate dibasic 3 parts, 2 parts, sodium-chlor, glucose 28 parts, 30 parts, fructose, 100 parts, brown sugar;
In order to achieve the above object, the preparation method for the solution shallow lake genus bacillus medicine fertilizer of scene, farm fermentation comprises following steps:
First by colistine sulfate 18 parts, tennecetin 1 part, ALANINE 12 parts, α-amino-isovaleric acid 22 parts, yeast leaching 100 parts, powder, peptone 100 parts, Sodium phosphate dibasic 3 parts, 2 parts, sodium-chlor, glucose 28 parts, 30 parts, fructose, 100 parts, brown sugar, places in sealing stirred pot and stirs 20-30 minute, mix, then after being added by 90 parts, Paenibacillus polymyxa bacterium powder and stirring in stirred pot, packing 500 grams is often wrapped, vacuumize packaging, the Paenibacillus polymyxa medicine namely obtained for scene fermentation is fertile;
Wherein, the application of described a kind of Paenibacillus polymyxa medicine fertilizer for the fermentation of scene, farm, comprises the following steps:
the fertile method of carrying out fermenting at scene, farm of this Paenibacillus polymyxa medicine is: the water-wet first the Paenibacillus polymyxa medicine of above-mentioned preparation fertilizer being added its 3-5 times weight is mixed well, water is supplied again after 30 minutes, to reach the ratio that Paenibacillus polymyxa medicine ratio that is fertile and water is 1:20, after adjusting PH to 8.2, start to cultivate, within first 12 hours, per minute air flow liquid-gas ratio is 1:0.6, after 12 hours, air flow is 1:1, culture temperature is 20-35 DEG C, cultivate after 16-24 hour, the fermented liquid of the Paenibacillus polymyxa medicine fertilizer of on-the-spot fermentation can be obtained.
Embodiment 4
Utilize the application of on-the-spot fermented liquid on plastic tent cucumber of embodiment 3 Paenibacillus polymyxa medicine fertilizer
Experimental field in certain large-scale plantation cooperative society of Huludao City, Liaoning, its plastic tent cucumber needs the problem solved to have following on producing: when A. kind is planted, fertilizer does not become thoroughly decomposed, and causes the problem of smoked canopy; B. the plot pathogenic bacteria accumulation of many years continuous cropping, causes the problem of the large generation of disease; C. soil compaction, root system development is bad, the problem that flower is pinched; D. in soil there is the problem that serious problem etc.E. improves utilization rate of fertilizer in root knot nematode;
Case study: use fertilizer, agricultural chemicals, hormone etc. to pollute soil, cause soil surface characters to accumulate, probiotics reduces, and Soil structure is unreasonable, soil compaction, have impact on the growth of cucumber root.If fundamentally do not solve the purifying problem of soil, control table and do not effect a permanent cure, on the contrary vicious cycle, more have impact on the quality of product;
Take measures: during nursery, can plastic shed soil be processed, the on-the-spot fermented liquid 10 kgs/acre of Paenibacillus polymyxa medicine fertilizer carries out turning over once after executing, and after field planting, filling with root ampoule is 2 kgs/acre, execute double as punching fertilising punching after the Spring Festival, each 2 kgs/acre;
Test-results: effect highly significant, mainly contains the effect of the application of following several respects:
A, it can eliminate stink, reduces ammonia fumigating seedling;
B, activating soil, improve the utilization ratio of soil and fertilizer;
C, shorten becoming thoroughly decomposed the time of fertilizer, shorten about 5 days, can early field planting, realizing restraining the growth of seedlings in seedling stage becomes possibility, and seedlings root is flourishing healthy and strong, improves the resistance against diseases of seedling;
D, the quantity of probiotics can be increased, suppress the accumulation of pathogenic bacteria; Effectively can control the generation of soil-borne disease, especially to root rot, root knot nematode disease, damping-off, bacterial wilt has extraordinary preventive effect;
E, provide comfortable edatope to cucumber growth, make plant strain growth healthy and strong, prolong growth period, last year, plastic tent cucumber also grew vigorous to mid-August, and cucumber yield is high, than contrast volume increase more than 25%; Product are of fine quality, profitable simultaneously, and mu annual income can reach more than 50,000 yuan;
F, minimizing agricultural chemicals and fertilizer amount, purification soil, reduces costs, and has walked out the way of benign cycle Sustainable development, has had far-reaching realistic meaning and historic significance.
Claims (5)
1. the Paenibacillus polymyxa medicine fertilizer for the fermentation of scene, farm, it is characterized in that, the raw material preparing this Paenibacillus polymyxa medicine fertilizer is: Paenibacillus polymyxa bacterium powder 50-200 part, colistine sulfate 10-50 part, tennecetin 1-2 part, ALANINE 3-40 part, α-amino-isovaleric acid 8-50 part, yeast leaching powder 50-200 part, peptone 50-200 part, Sodium phosphate dibasic 1-10 part, sodium-chlor 1-10 part, glucose 20-200 part, fructose 20-50 part, brown sugar 20-200 part.
2. a kind of Paenibacillus polymyxa medicine for the fermentation of scene, farm according to claim 1 is fertile, it is characterized in that, its preparation method is: first by colistine sulfate 10-50 part, tennecetin 1-2 part, ALANINE 3-40 part, α-amino-isovaleric acid 8-50 part, yeast leaching powder 50-200 part, corn starch 50-200 part, Sodium phosphate dibasic 1-10 part, sodium-chlor 1-10 part, fructose 20-50 part, brown sugar 20-200 part, place in sealing stirred pot and stir 20-30 minute, mix, again Paenibacillus polymyxa bacterium powder 50-200 part is added after stirring in stirred pot, packing 500 grams is often wrapped, vacuumize packaging, namely the Paenibacillus polymyxa medicine obtained for scene fermentation is fertile.
3. the application of a kind of Paenibacillus polymyxa medicine fertilizer for the fermentation of scene, farm according to claim 1, is characterized in that, comprise the following steps:
the fertile method of carrying out fermenting at scene, farm of this Paenibacillus polymyxa medicine is: the water-wet that the Paenibacillus polymyxa medicine fertilizer first claim 2 prepared adds its 3-5 times weight is mixed well, water is supplied again after 30 minutes, to reach the ratio that Paenibacillus polymyxa medicine ratio that is fertile and water is 1:10 to 1:30, after adjusting PH to 8-8.2, start to cultivate, within first 12 hours, per minute air flow liquid-gas ratio is 1:0.6, after 12 hours, air flow is 1:1, culture temperature is 20-35 DEG C, cultivate after 16-24 hour, the fermented liquid of the Paenibacillus polymyxa medicine fertilizer of on-the-spot fermentation can be obtained,
the application of Paenibacillus polymyxa medicine fertilizer for the fermentation of scene, farm be the fermented liquid of the Paenibacillus polymyxa medicine fertilizer of on-the-spot fermentation in arable farming to soil-borne disease, the application of leaf diseases;
the application of Paenibacillus polymyxa medicine fertilizer for the fermentation of scene, farm be the fermented liquid of the Paenibacillus polymyxa medicine fertilizer of on-the-spot fermentation in arable farming as base fertilizer, topdress and foliage fertilizer and the application of spreading manuer in holes and dressing seed;
Wherein, for soil-borne disease as tobacco bacterial wilt, balck shank, watermelon blight, cucumber fusarium axysporum etc., by step
the fermented liquid dilution 500-800 of preparation doubly after adopt the method for filling with root to prevent and treat, when transplanting and their early stage medication, each is 0.5-0.8 kg/acre by fermentation liquid measure;
Wherein, for leaf diseases as rice blast, rice sheath blight disease, powdery mildew of strawberry etc., by step
the fermented liquid dilution 200-500 of preparation doubly adopts the method for spraying to prevent and treat, and starts medication at the initial stage of a disease, uses once every 7 days, 3-5 time altogether;
Wherein, during as base fertilizer, by step
fermented liquid and other fertilizers of preparation and chemical fertilizer with the use of, be 5-20 kg/acre by fermentation liquid measure at every turn;
Wherein, as when topdressing, by step
the fermented liquid dilution 500-1000 of preparation doubly fills with root, and each is 1-3 kg/acre by fermentation liquid measure;
Wherein, as foliage fertilizer, by step
the fermented liquid dilution 300-800 times of foliage-spray of preparation, till positive and negative blade face all squirts, each is 0.3-0.5 kg/acre by fermentation liquid measure;
Wherein, during for dressing seed, by step
doubly, dress seed the fermented liquid dilution 200-500 of preparation 2-12 hour.
4. a kind of Paenibacillus polymyxa medicine for the fermentation of scene, farm according to claim 1 is fertile, it is characterized in that the spore content of described Paenibacillus polymyxa bacterium powder is not less than 10,000,000,000 cfu/g.
5. a kind of Paenibacillus polymyxa medicine for the fermentation of scene, farm according to claim 1 is fertile, it is characterized in that the water content of described each component raw material is lower than 5%.
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| CN107345215A (en) * | 2017-07-27 | 2017-11-14 | 上海万力华生物科技有限公司 | One plant of Paenibacillus polymyxa and its application in powdery mildew of cucumber and downy mildew preventing and treating |
| CN109832273A (en) * | 2019-03-08 | 2019-06-04 | 浙江大学 | The pesticide control of application and rice disease of the branched-chain amino acid in the pesticide control of preparation rice disease |
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| CN111533602A (en) * | 2020-04-24 | 2020-08-14 | 安徽汉合生物科技有限公司 | Special foliar fertilizer for selenium-rich biological rice |
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| CN105734000A (en) * | 2016-05-06 | 2016-07-06 | 南京农业大学 | Paenibacillus polymyxa NSY50 with capabilities of promoting growth and preventing diseases |
| CN107345215A (en) * | 2017-07-27 | 2017-11-14 | 上海万力华生物科技有限公司 | One plant of Paenibacillus polymyxa and its application in powdery mildew of cucumber and downy mildew preventing and treating |
| CN107345215B (en) * | 2017-07-27 | 2020-07-03 | 上海万力华生物科技有限公司 | Paenibacillus polymyxa and application thereof in prevention and treatment of powdery mildew and downy mildew of cucumber |
| CN109832273A (en) * | 2019-03-08 | 2019-06-04 | 浙江大学 | The pesticide control of application and rice disease of the branched-chain amino acid in the pesticide control of preparation rice disease |
| CN109832273B (en) * | 2019-03-08 | 2020-04-21 | 浙江大学 | Application of branched chain amino acid in preparation of rice disease control agent |
| CN110483111A (en) * | 2019-09-12 | 2019-11-22 | 广西垂青生物科技有限公司 | A kind of biological organic fertilizer and preparation method and application for preventing and treating soil-borne disease |
| CN111533602A (en) * | 2020-04-24 | 2020-08-14 | 安徽汉合生物科技有限公司 | Special foliar fertilizer for selenium-rich biological rice |
| CN111620745A (en) * | 2020-05-09 | 2020-09-04 | 大连理工大学 | Method for producing bio-organic fertilizer by degrading agricultural wastes with biological agent |
| CN111620745B (en) * | 2020-05-09 | 2021-08-24 | 大连理工大学 | Method for producing bio-organic fertilizer by degrading agricultural wastes with biological agent |
| CN113930466A (en) * | 2021-10-28 | 2022-01-14 | 吉林大学 | Paenibacillus-like genistein and preparation method thereof |
| CN113930466B (en) * | 2021-10-28 | 2023-12-08 | 吉林大学 | Paenibacillus genistein and preparation method thereof |
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