CN101671210A - Preparation method of paecilomyces lilacinus biological fertilizer - Google Patents
Preparation method of paecilomyces lilacinus biological fertilizer Download PDFInfo
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Abstract
The invention provides a preparation method of paecilomyces lilacinus biological fertilizer, belonging to the technical field of agrobiomechanics, and solving the problems in the prior art. The preparation method comprises the steps of: taking the paecilomyces lilacinus as bacterial strain, inoculating triangular flask liquid bacterial strain, and fermenting with czapek-dox medium to prepare liquid bacterial strain for production; taking decomposed or semi-decomposed fermenting bed padding materials in a culturing farm as basic medium, matching with accessorial carbon source and water, inoculating the liquid bacterial strain, and fermenting in a ventilation way in the depth layer of a solid fermenting pond to prepare paecilomyces lilacinus solid fermentation culture material; and drying the paecilomyces lilacinus solid fermentation culture material and grinding. The invention takes high-effective eelworm biocontrol bacteria paecilomyces lilacinus as the bacterial strain, takes the decomposed or semi-decomposed fermenting bed padding materials in the culturing farm as main raw materials, and ferments to prepare the paecilomyces lilacinus biological fertilizer to be taken as soil microorganism fertilizer which can prevent and cure the parasitic eelworm and various plant pathogenic bacteria, promote the plant growth, and reduce the pesticide residue in the soil.
Description
Technical field
The present invention relates to the fertile medicine of the dual-purpose soil microorganisms of fertile medicine, more specifically relate to a kind of preparation method of paecilomyces lilacinus biological fertilizer.
Background technology
In traditional agriculture is produced, for increasing both production and income, peasant household often uses products such as chemical fertilizer and production hormone to promote plant growth in a large number, use agricultural chemicals control disease and pest simultaneously, but because these chemical fertilizer, agricultural chemicals and production hormone etc. nearly all are that contemporaneity uses, supplementary additives such as dimethylbenzene wherein etc. are repeatedly residual, make the Soil structure in many places, land fertility and plant ecological are unbalance, take place to degenerate and mutation, cause particularly vegetables of farm crop, the intractable disease appears in fruit trees etc., and administers the pollution that the used chemical pesticide of these diseases has increased soil and agricultural-food.In this case, some producers begin to research and develop the preparation that has drug effect and fertilizer efficiency simultaneously, to reduce the amount of application of chemical fertilizer and agricultural chemicals, the saving cost that reaches, the purpose of reducing work hours, reduce Land pollution.These preparations can be divided into again in the mode of medicine and use and have concurrently the fertile medicine of fertilizer efficiency and the medicine fertilizer that uses and have concurrently drug effect with the form of fertilizer, normally simply agricultural chemicals and fertilizer are filled a prescription according to a certain percentage and mix mutually, and the agricultural chemicals of producing by certain technology and the recombiner of fertilizer, both bring into play its effectiveness separately separately, but between there is no interaction.
Nemas is the class root disease that the coelenterates nematode causes, has a strong impact on crop yield.At present, nemas takes place serious day by day, and nearly all crop all has eelworm harm.Parasitic nematode becomes one of important pathogen of plant infectious diseases, and agriculture production has a significant impact to the whole world.According to estimates, the whole world is approximately 12.3% because of the loss that eelworm harm causes to grain and fibre crops every year, causes about 1,000 hundred million dollars loss.China's proportion of crop planting industry is lost nearly 23,300,000,000 yuan because of the harm that nematode is caused every year.In the control of nematode most widely used general be to adopt nematocides to carry out chemical prevention, but the common toxicity of nematocides is higher, can increase the pollution of environment, more and more scholars recognizes that the control parasitic nematode must be taked aggregate measures or adopt with the biological and ecological control is the sustainable governing system of core.Fungi is the more natural enemy biology of research in the biocontrol of nematodes, also is one of of paramount importance biological and ecological methods to prevent plant disease, pests, and erosion factor.Wherein famous, also be that a most important opportunistic fungus Paecilomyces lilacinus (Paecilomyces lilacinus (Thom) Samson) is distributed widely in all over the world, easily from soil, fungi sclerotium separates obtaining, and easily cultivates on nematode and the insect.
Chinese scholars has been carried out a large amount of research to this bacterium, facts have proved for many years: the Paecilomyces lilacinus effect is remarkable, and has long-lasting; Multiple nematode is prevented and treated usefulness, is the most promising biological prevention and control agent of control parasitic nematode.Simultaneously, this bacterium has suitable resistance to the fertile medical instrument of killing of present use, can with kill fertile medicine and use with, improve preventive effect.With this bacterium commercialization, commodity are called Biocon in Philippines.
Studies show that, in the process of growth of Paecilomyces lilacinus, particularly in the submerged fermentation culturing process, this bacterium can produce abundant derivative, the similar indolylacetic acid product of the first, its the most significant physiological function is the growth that promotes root system of plant and plant when lower concentration, the effect that can also produce similar growth hormone and phytokinin in addition.Therefore using Paecilomyces lilacinus at root system can not only obviously suppress nematode infection, and can promote the growth of plant nutrition organ, and the sprouting to seed simultaneously also has promoter action with growth.
Paecilomyces lilacinus has antagonistic activity to the various plants pathogenic bacteria, there are some researches show: Paecilomyces lilacinus bacterial strain 36-1 has remarkable antagonistic activity to 8 kind of plant pathogenic bacterias, they are corn southern leaf blights, wheat scab, cotton wilt, the water rice bakanae disease, rice sheath blight disease, citrus pathogens penicillium, citrus green mold bacterium, cucumber anthracnose, Abdul (2000) utilizes Paecilomyces lilacinus control cotton bacterialo wilt disease and cotton virus disease, the result shows that paecilomycerol can obviously reduce cotton (NIAB-18, the CIM-240 kind) viral disease, and effectively suppress bacterialo wilt disease, also from the greenhouse, filter out high antagonism power bacterial strain, be sprayed on the wheat leaf blade, in order to the control speckled leaf blotch.Correlative study also confirms: Paecilomyces lilacinus can significantly reduce the sickness rate of Sunflower Receptacle and natto root knot, and can suppress the tomato root rot that caused by Fusarium.
Discover that Paecilomyces lilacinus can promote the dissolving of difficultly-soluble phosphates, through laboratory test: the Paecilomyces lilacinus bacterium to the correction degradation rate of Volaton about 94.Similar research shows that also Paecilomyces varioti can also promote the decomposition of many chemical polymerization things (as organophosphorus pesticide, leather-making waste water etc.).
In sum, Paecilomyces lilacinus not only can be developed becomes outstanding wireworm-killing biologic agricultural chemicals, and can prevent and treat plurality of plant diseases simultaneously, and also can be used as efficiently, bio-feritlizer uses, reduce the residual of chemical pesticide and chemical fertilizer, promote the production of non-polluted farm product.
Summary of the invention
The preparation method who the purpose of this invention is to provide a kind of paecilomyces lilacinus biological fertilizer; solve in the prior art as only medicine and fertilizer are simply mixed; medicinal and fertile all not good with effect; a series of problems of contaminate environment or the like; preparation process is simple; the bacterium activity is good in the paecilomyces lilacinus biological fertilizer of cultivating; when solid fermentation is cultivated directly with fertilizer as substratum; and with microorganism as medicinal ingredients; make fertile medical instrument of the present invention that the combination of medicinal and fertile usefulness be arranged; the equal highly significant of fertilizer efficiency drug effect, and environmentally safe have remarkable economic efficiency and environmental protection benefit.
Technology contents of the present invention: the preparation method of paecilomyces lilacinus biological fertilizer, step comprises:
(1) production prepares with liquid spawn: be that bacterial classification is made second-class liquid isolate with the Paecilomyces lilacinus, and serve as to produce to use substratum with liquid spawn with Cha Shi liquid nutrient medium or PDA liquid nutrient medium, at fermentor tank inoculation second-class liquid isolate, cultivate to produce and use liquid spawn;
(2) configuration solid fermentation substratum, liquid spawn is used in the production of inoculation step (1) gained, changes the solid fermentation pond over to, and the thick-layer ventilating fermentation prepares the Paecilomyces lilacinus solid fermentation culture;
(3) with the Paecilomyces lilacinus solid fermentation culture of step (2), 50～80 ℃ of oven dry are pulverized, and promptly get the fertile medicine of Paecilomyces lilacinus.
Excellent results of the present invention is: select for use efficient nematode biocontrol microorganisms Paecilomyces lilacinus for producing bacterial strain, not only plant parasitic nematodes had high virulence, and the various plants pathogenic bacteria had antagonistic activity, simultaneously can also produce class tethelin material, promote plant-growth, the fertilizer efficiency effect that further improves product; As the basic medium of Paecilomyces lilacinus solid fermentation, plant's fermenting bed padding that becomes thoroughly decomposed itself promptly is good fertilizer source, also can better grow the condition that provides surely in soil for bacterial strain when providing fertility for crop.Potted plant and field test results shows, and is remarkable to crop root knot nematode disease evil prevention effect, and good disease-resistant growth-promoting effect is arranged simultaneously; Preparation process is simple, biocontrol microorganisms spore production rate height, active good in the paecilomyces lilacinus biological fertilizer of cultivation; Solid fermentation is cultivated directly with fertilizer as basic medium; with efficient nematode biocontrol microorganisms Paecilomyces lilacinus as medicinal ingredients; make fertile medical instrument of the present invention that the combination of drug effect and fertilizer efficiency be arranged; support medicine with fertilizer, with the medicine getting fat, both mutually promote; the equal highly significant of fertilizer efficiency drug effect; can reduce the usage quantity of chemical pesticide and chemical fertilizer, environmentally safe has remarkable economic efficiency and environmental protection benefit.
Embodiment
The Paecilomyces lilacinus that the present invention sells with the grand monarch of the section agricultural science and technology company limited in Fujian serves as to produce bacterial strain, the preparation paecilomyces lilacinus biological fertilizer, and each step specifies as follows:
Strain preparation:
Second-class liquid isolate is cultivated: this step can be cultivated by Paecilomyces lilacinus second-class liquid isolate routine techniques, also can cultivate according to following concrete steps:
A. the solid slant strains is cultivated: configuration Cha Shi solid medium or PDA solid medium, and sterilization, the streak inoculation Paecilomyces lilacinus places in the constant incubator and cultivated 7～9 days under 25～30 ℃;
The Cha Shi substratum that described solid slant strains is used in cultivating: NaNO
33g, K
2HPO
41g, KCl 0.5g, MgSO
47H
2O 0.5g, FeSO
40.01g, sucrose 30g, agar 15～20g, H
2O 1000mL, the pH nature;
The PDA solid medium that described solid slant strains is used in cultivating: take by weighing the 200g potato, clean peeling is cut into small pieces, and adds water 1000mL and boils half an hour or steaming and decocting under high pressure 20 minutes, filtered through gauze, add 10～20g glucose and 17～20g agar again, fully filtered through gauze while hot after the dissolving.
B. second-class liquid isolate is cultivated: identical except that not adding the agar all the other compositions or preparation method in second-class liquid isolate culture medium prescription and the described solid slant strains culture medium prescription, the second-class liquid isolate substratum is sub-packed in being sub-packed in the triangular flask of 1L by the amount of 250mL/ bottle, after the sterilization cooling, on the aseptic technique platform, collude from the Paecilomyces lilacinus that the solid slant culture base is preserved with transfering loop and to get conidium one ring, be seeded in the triangular flask in the liquid nutrient medium, shaking table 110～150rpm, 25～30 ℃ cultivated 4～5 days.
Described cultivation is produced with the Cha Shi liquid nutrient medium of using in the liquid spawn or PDA liquid culture based on after tinning in the airlift fermentor, sterilizing, press the amount inoculation second-class liquid isolate of substratum 1wt%, control air flow 1: 1,25～30 ℃ of temperature, fermentation culture 7～8 days, liquid spawn is used in the production that obtains Paecilomyces lilacinus.
Production is used substratum with liquid spawn:
Cha Shi liquid culture based formulas is: NaNO
33g, K
2HPO
41g, KCl 0.5g, MgSO
47H
2O 0.5g, FeSO
40.01g, sucrose 30g, H
2O 1000mL, the pH nature;
The PDA liquid nutrient medium is: take by weighing the 200g potato, clean peeling is cut into small pieces, and adds water 1000mL and boils half an hour or steaming and decocting under high pressure 20 minutes, and filtered through gauze adds 10～20g glucose again, fully filtered through gauze while hot after the dissolving.
(2) the thick-layer ventilating fermentation prepares the Paecilomyces lilacinus solid fermentation culture
Basic medium 60～80 weight parts, auxiliary carbon source 20～40 weight parts, the Hou Jiashui that stirs, the adjustment water ratio is 30～50wt%, the production of inoculation step (1) is with liquid spawn 10～15 weight parts, change in the solid fermentation pond, ventilate,, stirred once in 24～30 hours at 25～30 ℃ of bottom fermentations, fermented 7～9 days, and promptly obtained the solid fermentation culture of Paecilomyces lilacinus.
Basic medium is the organic bedding and padding of plant's fermentation bed that become thoroughly decomposed or partly become thoroughly decomposed; Auxiliary carbon source is one or more of rice bran, wheat bran, W-Gum or straw powder.
The solid fermentation culture of this Paecilomyces lilacinus, spore count can reach 500,000,000/g dry product.
(3) the Paecilomyces lilacinus solid fermentation culture that step (2) is obtained is after 50～80 ℃ of dryings, pulverizing, quality inspection, packaging final prod.
Below in conjunction with specific embodiment, further set forth the present invention.These embodiment only are used to the present invention is described and are not used in and limit the scope of the invention.The experimental technique of unreceipted actual conditions among the embodiment, condition routinely usually, or the condition of advising by manufacturer.Under the prerequisite of not doing to indicate especially, all involved umbers are mass fraction, and all per-cents are mass percent.
Embodiment 1
(1) preparation of the second-class liquid isolate of Paecilomyces lilacinus, the concrete operations step is as follows:
A. slant strains (solid)
Take by weighing 20 portions of potatos, clean peeling is cut into small pieces, add 100 parts in water and boiled half hour or steaming and decocting under high pressure 20 minutes, filtered through gauze adds 2 parts of glucose and 2 parts of agar again, fully filtered through gauze while hot after the dissolving, the packing test tube, take out test tube pendulum inclined-plane after 15 pounds of steams (121 ℃) were sterilized 20 minutes, cooling back streak inoculation Paecilomyces lilacinus NH-PL-03 bacterial strain places thermostat container to cultivate 8 days for interior 28 ℃.
B. second class inoculum (liquid)
Culture medium prescription is sub-packed in the triangular flask with the inclined-plane but do not add agar, after the sterilization cooling, on the aseptic technique platform, collude from the Paecilomyces lilacinus that the solid slant culture base is preserved with transfering loop and to get conidium one ring, be seeded in the liquid nutrient medium, shaking table 110rpm, 28 ℃ cultivated 4 days.
Produce preparation with liquid spawn
Substratum: Cha Shi liquid nutrient medium, NaNO
33 parts, K
2HPO
41 part, 0.5 part of KCl, MgSO
47H
20.5 part of O, FeSO
40.01 part, 30 parts of sucrose, H
21000 parts of O, the pH nature.
After airlift fermentor tinning, the sterilization, 50 parts of inoculation second-class liquid isolates, control air flow 1: 1,28 ℃ of temperature, fermentation culture 8 days, liquid spawn is used in the production that obtains Paecilomyces lilacinus.
(2) the thick-layer ventilating fermentation prepares the Paecilomyces lilacinus solid fermentation culture
80 parts of the plant's fermenting bed paddings that partly becomes thoroughly decomposed, 20 parts in wheat bran, the Hou Jiashui that stirs, adjusting water ratio is 40%, the production of inoculation step (1) is with 10 parts of liquid spawns (more than be weight part), change in the solid fermentation pond, ventilate,, stirred once in 24 hours at 28 ℃ of bottom fermentations, fermented 8 days, and promptly obtained the solid fermentation culture of Paecilomyces lilacinus.
The solid fermentation culture of this Paecilomyces lilacinus, spore count can reach 500,000,000/g dry product.
(3) with the Paecilomyces lilacinus solid fermentation culture of step (2), 50～80 ℃ of oven dry are pulverized, and promptly get the fertile medicine of Paecilomyces lilacinus.
Embodiment 2: the preparation of the fertile medicine of Paecilomyces lilacinus
The step different with embodiment 1 is as follows, and all the other unaccounted steps are with embodiment 1:
Step (2) thick-layer ventilating fermentation prepares the Paecilomyces lilacinus solid fermentation culture
70 parts of the plant's fermenting bed paddings that becomes thoroughly decomposed, 15 parts in rice bran, 15 parts of W-Gums, Hou Jiashui stirs, adjusting water ratio is 40%, the production of inoculation step (1) changes in the solid fermentation pond with 8 parts of liquid spawns (more than be weight part), ventilates, at 27 ℃ of bottom fermentations, stirred once in 24～30 hours, fermented 9 days, promptly obtain the solid fermentation culture of Paecilomyces lilacinus.
The effect test of the fertile medicine of Paecilomyces lilacinus of the present invention's preparation
The potted plant control test of tobacco root knot nematode
1 for collection of examination nematode and breeding
Obtain from the rhizosphere that nematode infects for the examination nematode.The root knot of being injured identifies at test under microscope and through scanning electron microscopic observation, confirm as javanese root knot nematode (Meloidogyme javanica) after, adopt the egg capsule inoculation method, nematode is inoculated in cigarette shoot root portion carries out expanding propagation.
2 tests are handled
Add earlier in the cultivation alms bowl through the Exposure to Sunlight sterilization and through separating the strong soil that confirms no nematode, do following 4 kinds of processing: 1. fertile medicine is handled: every alms bowl adds rhizosphere sick soil and the 50g fertile medicine of 100g through inoculation reproduction band root knot; 2. nematocides is handled: every alms bowl adds the sick soil and the 2g nematocides (3% carbofuran Granules) of 100g band root knot, is executing 1 nematocides (every alms bowl 2g) behind 15d; 3. Exposure to Sunlight disinfection soil: every alms bowl adds the soil of 100g through sunning sterilization, not wiring worm, not dispenser.Contrast (CK): every alms bowl adds the sick soil of 100g band root knot, does not does not prevent and treat.Every treatment group is all implanted the uniform three leaf age cigarette seedlings of growing way, every processing 20 alms bowls, and test repeats 2 times.The pot experiment bacteria using amount is the every strain of 50g/.
After the field planting, observe the growing state of plant.Behind the tobacco seedling growth 2 months, extract the cigarette seedling, remove root soil, measure plant height one by one, leaf is long, weighs up every strain overground part and root system fresh weight with general utility balance with flushing with clean water.Carry out the classification of root knot severity by following standard: 0 grade for all root systems all not by nematode infection, no root knot; The II level is the 26%-50% that root knot accounts for total root system; The III level accounts for the 51%-75% of total root system for the root knot number; The IV level accounts for the 76%-100% of total root system for the root knot number.Disease index=strain numbers at different levels * total strain the number of severity progression/investigation * the highest sick progression
Calculate disease index in each treatment group by above formula.Every alms bowl is got 1 part of root system soil sample, by the treatment group mixing, takes by weighing 50g, examines under a microscope, count in the rhizosphere soil 2 age the nematode population level.
3 prevention effect
3.1 drug effect
The tobacco root knot nematode causes the overground part poor growth in the tobacco root generation root knot that causes harm.Test-results shows, uses fertile medicine and nematocides and all can suppress the breeding of root knot nematode effectively and alleviate to cause harm.The nematocides effect is the strongest, tobacco root knot nematode disease disease index comparison according to descend 39.5%, two age the nematode number reduce 79.2%, secondly be to use fertile medicine, the comparison of plant disease index according to descended 34.0%, two age the nematode number reduced by 76.3% (table 1)
Two nematode numbers in age (in the 50g rhizosphere soil) in table 1 disease index and the soil
3.2 fertilizer efficiency
Test-results shows that the fertile medicine of Paecilomyces lilacinus not only plays restraining effect to nematode but also can promote the tobacco plant growth.Though compare with nematocides, the root knot disease index is high slightly, and plant strain growth is than the red treatment group stalwartness of furans.From the plant growing way, the cigarette strain leaf look that adds fertile medicine is dark green, grows fine; Add the plant base of leaf cripetura of the tobacco of 3% carbofuran Granules, leaf colour cast Huang, plant height are starkly lower than the cigarette strain that adds fertile medicine; The contrast that only adds sick soil is then obviously short and small, and growing way is poor; Use the cigarette strain robust growth of Exposure to Sunlight disinfection soil plantation.Cigarette strain growth traits measurement result shows that the average fresh weight of treated tobacco plant and root system, plant height, Ye Changjun are apparently higher than untreated plant.The average fresh weight comparison of plant overground part of using fertile medicine is according to increasing by 96.25%, and root system increases by 153.85%, and plant height and leaf are long to increase by 75.89% and 38.89% respectively, the most remarkable with the root system weightening finish.In addition, use average fresh weight, plant height and the Ye Changjun of its overground part of cigarette strain of fertile medicine and root system apparently higher than the cigarette of using nematocides.To use fertile medicine group and use the strong native treatment group of Exposure to Sunlight sterilization to compare, the overground part of fertile medicine group and root system fresh weight 2,7 all " Invest, Then Investigate "s are found: 2 nematode minimizings in age 76.3% in the soil, root knot index decreased 43.6%, cigarette strain root growth amount increases by 153.7%, and the over-ground part fresh weight increases by 44.4%
The control test of sub-district, eggplant root knot nematode field
The preparation of 1 root knot nematode oophyte:
After the root knot of indoor cultivation (band root knot nematode) is sheared, filter, under anatomical lens, calculate the root-knot nematode egg number with double gauze.
2 experiments are handled
Tested in 2003 and carry out in academy of agricultural sciences, Geling Hill Fujian, Yongtai, Fuzhou City, Fujian Province biotechnology center experiment base, establish three fertile medicines altogether and handle, every processing interts a contrast, and the every furrow in field are planted 36 strains, whenever be treated to three furrow and be divided into two rows, each 108 strain of promptly every processing.During two true leaves of eggplant length, three processing are converted the clear water pouring with the fertile medicine of Paecilomyces lilacinus respectively.Water bacterium one week back with the inoculation of root knot nematode oophyte, 500 strain eggplant seedlings connect altogether 4000 ovum (10 of suspension concentration ≈/mL), two week the back repeat last time and connect ovum to handle.Behind the eggplant transplantation of seedlings land for growing field crops, respectively water once (spore concentration ≈ 1.2 * 10 of fertile medicine at vegetative period, florescence respectively
7Individual/mL), CK waters with clear water.Quantity of tracking investigation root knot nematode and plant strain growth situation are estimated the effect of fertile medicine.
3 prevention effect
3.1 drug effect
Table 1. field eggplant rhizosphere root knot nematode population density sample survey (unit: bar/50g soil)
| Before florescence | ??12 | ??78 | ??30 | ??88.44 |
| Fruiting period | ??0 | ??18 | ??24 | ??95.96 |
| CK (fruiting period) | ??276 | ??132 | ??137 | ??47.50 |
| Radix | ??486 | ??234 | ??318 | ??/ |
Investigation result sees Table 1, and in a week after the dispenser, the fertile medicine of Paecilomyces lilacinus is handled the sub-district root knot nematode insect population rate that goes down and descended 58.02%, 20.51%, 66.04% respectively, reaches 88.44 and 95.96% with the fruiting period average population rate of going down before the florescence.The field nature insect population rate that goes down also reaches 47.50%, and the biological and ecological methods to prevent plant disease, pests, and erosion factor that also has other root knot nematode in the field soil is described.
3.2 fertilizer efficiency
Week sampling is observed behind the medicine, handle the eggplant root system for three kinds and infected in various degree by root knot nematode, and aggregate level is lower, compares the fertile medicine of Paecilomyces lilacinus and handles, and the CK root nodule is obviously big.Get the fruiting period plant and observe (Fig. 2), the Paecilomyces lilacinus agent is handled better in the plant strain growth The Characters; On root was caused harm shape, the eggplant plant root elegans development that the fertile medicine of Paecilomyces lilacinus is handled was slow, root nodule is tiny, well developed root system, and fibrous root is more, and the contrast strain shows as that root swells obviously, root nodule is bigger, the also little prosperity of root system.
The output that the sub-district is respectively handled in the field sees Table 2, and the sub-district stimulation ratio of using Paecilomyces lilacinus is between 24.32%-29.56%, and average stimulation ratio reaches 26.60%,
Ultimate production is respectively handled to stimulation ratio when in table 2 field
Claims (7)
1. the preparation method of a paecilomyces lilacinus biological fertilizer, it is characterized in that: described preparation method's step comprises:
(1) production prepares with liquid spawn: be that bacterial classification is made second-class liquid isolate with the Paecilomyces lilacinus, and serve as to produce to use substratum with liquid spawn with Cha Shi liquid nutrient medium or PDA liquid nutrient medium, at fermentor tank inoculation second-class liquid isolate, cultivate to produce and use liquid spawn;
(2) configuration solid fermentation substratum, liquid spawn is used in the production of inoculation step (1) gained, changes the solid fermentation pond over to, and the thick-layer ventilating fermentation prepares the Paecilomyces lilacinus solid fermentation culture;
(3) with the Paecilomyces lilacinus solid fermentation culture of step (2), 50～80 ℃ of oven dry are pulverized, and promptly get the fertile medicine of Paecilomyces lilacinus.
2. the preparation method of paecilomyces lilacinus biological fertilizer according to claim 1, it is characterized in that: the solid fermentation culture of described step (2) prepares specific as follows:
Basic medium 60～80 weight parts, auxiliary carbon source 20～40 weight parts, the Hou Jiashui that stirs, the adjustment water ratio is 30～50wt%, the production of inoculation step (1) is with liquid spawn 10～15 weight parts, change in the solid fermentation pond, ventilate,, stirred once in 24～30 hours at 25～30 ℃ of bottom fermentations, fermented 7～9 days, and promptly obtained the solid fermentation culture of Paecilomyces lilacinus.
3. the preparation method of paecilomyces lilacinus biological fertilizer according to claim 2 is characterized in that: described basic medium is the organic bedding and padding of plant's fermentation bed that become thoroughly decomposed or partly become thoroughly decomposed.
4. the preparation method of paecilomyces lilacinus biological fertilizer according to claim 2, it is characterized in that: described auxiliary carbon source is one or more of rice bran, wheat bran, W-Gum or straw powder.
5. the preparation method of paecilomyces lilacinus biological fertilizer according to claim 1 is characterized in that: the second-class liquid isolate concrete steps are in the described step (1):
A. the solid slant strains is cultivated: configuration Cha Shi solid medium or PDA solid medium, and sterilization, the streak inoculation Paecilomyces lilacinus places in the constant incubator and cultivated 7～9 days under 25～30 ℃;
The Yao Shi solid culture based formulas that described solid slant strains is used in cultivating: NaNO
33g, K
2HPO
41g, KCl0.5g, MgSO
47H
2O 0.5g, FeSO
40.01g, sucrose 30g, agar 15～20g, water 1000mL, the pH nature;
The PDA solid medium that described solid slant strains is used in cultivating is: take by weighing the 200g potato, clean peeling is cut into small pieces, and adds water 1000mL and boils half an hour or steaming and decocting under high pressure 20 minutes, filtered through gauze, add 10～20g glucose and 17～20g agar again, fully filtered through gauze while hot after the dissolving;
B. second-class liquid isolate is cultivated: identical except that not adding the agar all the other compositions or preparation method in second-class liquid isolate culture medium prescription and the described solid slant strains culture medium prescription, the amount of second-class liquid isolate substratum by the 250mL/ bottle is sub-packed in the triangular flask of 1L, after the sterilization cooling, on the aseptic technique platform, collude from the Paecilomyces lilacinus that the solid slant culture base is preserved with transfering loop and to get conidium one ring, be seeded in the triangular flask in the liquid nutrient medium, shaking table 110～150rpm, 25～30 ℃ cultivated 4～5 days.
6. the preparation method of paecilomyces lilacinus biological fertilizer according to claim 1 is characterized in that: produce in the described step (1) with in the liquid spawn preparation process:
Described cultivation is produced with the Cha Shi liquid nutrient medium of using in the liquid spawn or PDA liquid culture based on after tinning in the airlift fermentor, sterilizing, press the amount inoculation second-class liquid isolate of substratum 1wt%, control air flow 1: 1,25～30 ℃ of temperature, fermentation culture 7～8 days, liquid spawn is used in the production that obtains Paecilomyces lilacinus.
7. according to the preparation method of claim 1 or 6 described paecilomyces lilacinus biological fertilizers, it is characterized in that: the production in the described step (1) is used substratum with liquid spawn:
Yao Shi liquid culture based formulas is: NaNO
33g, K
2HPO
41g, KCl 0.5g, MgSO
47H
2O0.5g, FeSO
40.01g, sucrose 30g, water 1000mL, pH nature;
The PDA liquid nutrient medium is: take by weighing the 200g potato, clean peeling is cut into small pieces, and adds water 1000mL and boils half an hour or steaming and decocting under high pressure 20 minutes, and filtered through gauze adds 10～20g glucose again, fully filtered through gauze while hot after the dissolving.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009103090441A CN101671210B (en) | 2009-10-29 | 2009-10-29 | Preparation method of paecilomyces lilacinus biological fertilizer |
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| CN103773697A (en) * | 2013-12-05 | 2014-05-07 | 江西天人生态股份有限公司 | paecilomyces lilacinus and application thereof |
| CN104016811A (en) * | 2014-05-09 | 2014-09-03 | 安徽福瑞德生物科技有限公司 | Beauveria bassiana biological compound medicine fertilizer and preparation method thereof |
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| CN105613591A (en) * | 2016-01-27 | 2016-06-01 | 刘宫介 | Formula and preparation method of environment improver capable of reducing quantity of pathogenic microorganisms |
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