CN101671212A - Microbial agent capable of reducing leaf vegetable plant diseases and insect pests and preparing method thereof - Google Patents
Microbial agent capable of reducing leaf vegetable plant diseases and insect pests and preparing method thereof Download PDFInfo
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Abstract
The invention relates to a microbial agent capable of reducing leaf vegetable plant diseases and insect pests and a preparing method thereof, belonging to the technical field of agricultural production application of microorganisms. The microbial agent capable of reducing leaf vegetable plant diseases and insect pests is characterized in that the microbial agent is prepared from the following rawmaterials: beauveria bassiana culture solution, streptomyces coelicolor culture solution, compound strain culture solution, garlic, molasses, Chinese medicinal herb and trace element inorganic salt solution, wherein the compound strain is composed of Bacillus subtilis culture solution, rhodopseudomonas palustris and saccharomycetes. The preparing method is as follows: respectively culturing beauveria bassiana and streptomyces coelicolor alone, culturing Bacillus subtilis, lactobacillus and saccharomycetes in a compound mode; mixing and fermenting three culturing products, Chinese medicinal herb, garlic and the like; and finally, adding trace elements into the compound fermentation product. The invention can obviously strengthen leaf vegetable organism disease resistance and resilience, improves soil and reduces soil-borne diseases.
Description
Technical field
The present invention relates to a kind ofly can reduce microbial preparation of leaf vegetable plant diseases and insect pests and preparation method thereof, can significantly strengthen leaf vegetables body disease-resistant power and resistance against disadvantage, improve the soil, reduce soil-borne disease, impel the volume increase of leaf vegetables safety; Effectively reduce the use of fertilizer and pesticide simultaneously, improve the quality of leaf vegetables, reduce planting cost, belong to microorganism in its application in agriculture field.
Background technology
Leaf vegetables can provide nutritive substances such as the carbohydrate that enriches, VITAMIN, Mierocrystalline cellulose for people, is indispensable food in the daily life.Yet, under the driving of the economic interests that covet, people have begun cruel activity in production of taking formula by force, bad means such as product for agriculture such as excessive applied nitrogen, agricultural chemicals, weedicide have been taked, the result causes serial problems such as the leaf vegetables quality worsens, disease and pest aggravates, pesticide residue exceeds standard, and has directly influenced leaf vegetables output and quality.Along with the raising of people's living standard and the reinforcement of concept of health, the ratio of leaf vegetables in people's diet formula is increasing, the quality safety of vegetables also becomes the problem of enduring concern to the fullest extent, and simultaneously, the excessive cultivation technique system of using chemical fertilizer, agricultural chemicals has also caused serious environmental problem.The safe yield-increasing technology of exploitation leaf vegetables disease control is extremely important.
Along with going deep into of research, formed the yield-increasing technology of different angles such as physics, chemistry, biology.Physical control utilizes the taxis insect-killing trapping more, reduces the leaf vegetables loss; Chemical terms, adopt used chemical fertilizer and the agricultural chemicals method of using low toxicity and low residue more, still can not avoid the residual gathering of hazardous and noxious substances fully; Biological angle then uses no or little chemical pesticide, improves the soil, suppresses disease and pest, promotes crop alimentary to absorb by useful microbial activities, reaches the effect of safety volume increase.But the research about the leaf vegetables yield-increasing technology focuses mostly in the physical chemistry field at present, and the relevant mature technology of biological field is less, and mostly is the application of single microorganism, and the vigor of bacterium is generally 10
6-10
8Cfu/ gram, function is also more single, and is few about the disease control of compound probiotic, safe yield-increasing technology.
Summary of the invention
The purpose of this invention is to provide and a kind ofly can significantly strengthen leaf vegetables body disease-resistant power and resistance against disadvantage, improve the soil, reduce soil-borne disease, impel complex microorganism preparations of leaf vegetables safety volume increase and preparation method thereof.
In order to achieve the above object, technical scheme of the present invention provides a kind of microbial preparation that can reduce leaf vegetable plant diseases and insect pests, it is characterized in that it is made by following raw material in weight part: beauveria bassiana nutrient solution 10-20, streptomycete nutrient solution 5-10, composite bacteria nutrient solution 15-30, garlic 1-15, molasses 20-40, herbal medicine 1-15 and micro-inorganic salt solution 0.2-0.8;
Wherein, described composite bacteria is made up of bacillus subtilis bacteria culture fluid 25-45wt%, Rhodopseudomonas palustris 30-50wt% and yeast 25-45wt%.
Described herbal medicine is that Radix Glycyrrhizae 30-50wt%, Japanese Honeysuckle 20-40wt% and Root of Indigowoad 20-30wt% form.
Described micro-inorganic salt solution is the inorganic salt solution that contains iron ion, molybdenum ion, cupric ion, boron ion and chlorion, above-mentioned ionic concn and be 1-5g/L.
The present invention also provides the above-mentioned preparation method that can reduce the microbial preparation of leaf vegetable plant diseases and insect pests, it is characterized in that, concrete steps are:
The first step: with first liquid nutrient medium 120-130 ℃ of sterilization after 15-30 minute, with tween-80 with weight ratio 100: 0.5-0.8 mixes; With beauveria bassiana by 1: the weight ratio of 80-100 be inoculated in the institute liquid nutrient medium mixture of winning in, at 25-35 ℃, oscillation frequency be under the condition of 110-130r/min in shaking table shaking culture 5-7d, sampling is observed, when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, stop fermentation, obtain the beauveria bassiana nutrient solution;
Described first liquid nutrient medium is made up of following raw material in weight part: analysis for soybean powder 2-4, peanut powder 2-5, sucrose 5-10, potassium primary phosphate 1.5-2, sal epsom 0.2-0.6, calcium chloride 0.01-0.02, manganous sulfate 0.001-0.008, VB8-10 and sterilized water 70-80;
Second the step: with second liquid nutrient medium 120-130 ℃ the sterilization 15-30 minute after, with streptomycete by 1: the weight ratio of 50-100 is inoculated in above-mentioned second liquid nutrient medium, regulating the pH value is 7.0-7.2, at temperature 25-30 ℃, oscillation frequency be under the condition of 200-240r/min in shaking table shaking culture 8-10d, sampling is observed, when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, stop fermentation, obtain the streptomycete nutrient solution;
Described second liquid nutrient medium is made up of following raw material in weight part: starch 10-15, Semen Maydis powder 8-10, yeast powder 2-5, soybean cake powder 2-5, lime carbonate 1.5-2.0, potassium primary phosphate 1.5-2, sal epsom 0.2-0.6, calcium chloride 0.01-0.02, cobalt chloride 0.0004-0.0007 and sterilized water 65-78;
The 3rd step: subtilis, Rhodopseudomonas palustris, yeast are pressed 25-45: 30-50: the weight ratio of 25-45 is mixed, by 1: the weight ratio of 50-100 is inoculated in the 3rd liquid nutrient medium, the pH value is adjusted to 6.8, after the 25-35 ℃ of speed with 150-300r/min stirs 15-25min, airtight standing for fermentation 2-4d, sampling is observed, and stops fermentation when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, obtains the composite bacteria nutrient solution;
Described the 3rd liquid nutrient medium is made up of following raw material in weight part: molasses 5-10, peptone 0.5-1, yeast powder 0.5-1, ammonium sulfate 0.2-0.5, sal epsom 0.2-0.5 and sterilized water 88-94;
The 4th step: garlic and herbal medicine are pulverized the back and beauveria bassiana nutrient solution, streptomycete nutrient solution, composite bacteria nutrient solution and molasses are mixed in proportion, insert the inherent 30-35 ℃ of fermentation 6-8d of closed tin, sampling is observed, treat that viable bacteria total concn number stops fermentation when surpassing 1,200,000,000/ml, obtain the mixed fermentation product;
The 5th goes on foot: go on foot in the mixed fermentation product that obtains the 4th and add micro-inorganic salt solution in proportion, obtain reducing the microbial preparation of leaf vegetable plant diseases and insect pests.
The temperature of shaking culture is 26-28 ℃ in described second step.
The used 5 kinds of bacterial classifications of the present invention are all bought from agriculture microbial strains preservation administrative center, and its preserving number is as follows: beauveria bassiana Beauveria bassiana (ACCC 30006); Streptomycete Streptomyces sp (ACCC 40069); Subtilis Bacillus subtilis (ACCC 10623); Rhodopseudomonas palustris Rhodosedoseudomousaspadustris (ACCC 10649); Yeast Saccharomyces cerevisiae (IFO 0203).
The principle of the invention is as follows:
The present invention adopts antimicrobial substance syntrophism fermentations such as multiple bacterial classification and garlic, herbal medicine, produces the metabolic substd of big content simultaneously, and viable bacteria content liquid is 1,200,000,000/mL.
The full muscardine of ball has the ability of objectionable impuritiess such as part degraded toluene compound and the ability of heavy metals such as bio-transformation cadmium and copper, the bag beauvericin of its generation in addition, the defense response that can suppress the insect hemocyte, it can be bred in host in a large number, destroy host's function of organization, the a large amount of accumulation of its meta-bolites oxalate in blood can make host's metabolic disturbance and death simultaneously.
Streptomyces can be carried out plant protection by antibiosis, competition and super parasitization in actinomycetales.Connect sterilization and can secrete antibiotics or extracellular ferment, can effectively suppress phytopathogen, small part can promote plant-growth or inducing plant to produce disease resistance; Also have stronger decomposition complexity and reach nitrogenous and nonnitrogenous organic ability, soil improvement is played an important role, can effectively reduce germ and grow, reach and prevent and treat the disease effect.
Rhodopseudomonas palustris can produce and discharge a kind of disease-resistant factor coenzyme in metabolic process, pathogenic bacterium in body and the soil are had certain restraining effect, also can with the actinomycetes symbiosis in the soil, make probiotics propagation, suppress other pernicious bacteria breeding, play ecological disease-proof functions.
Yeast flora is the cell protein content height not only, and in their growth metabolism process, can form multiple vitamin B group and physiologically active substance, help other microorganisms and vegeto-animal dietetic alimentation, strengthen the activity of effective bacterium, promote the performance of disease control function.
Use by microbial preparation, can in leaf vegetables root or soil, can produce and accumulate a large amount of nutritious microbial cells and useful metabolite, as organic acid, alcohol, aldehyde, VITAMIN, hormone, microbiotic, strengthen plant body vigor and metabolic capacity, impel root system and plant development, improve the chlorophyll content and the photosynthetic speed of photosynthetic area blade, promote the safety volume increase.
The mainly edible position of leaf vegetables is branches and leaves, needs to guarantee that its color and luster is dark green, if lack essential element, can cause the endogenous illness, therefore replenishes molybdenum, avoids plant to lack molybdenum and causes Nitrate Accumulation, causes maculopathy; Supplementation with copper is avoided plant to lack copper and is caused the electronic chain transmission to be obstructed, and plant wilt is thin and delicate.Boron make-up, the tip of a root and the shoot apical meristem necrosis of avoiding boron deficiency to cause.Replenish chlorine, avoid plant to lack chlorine and form leaflet and necrose.
Advantage of the present invention is as follows:
1. can strengthen the metabolism of plant, strengthen the stratum corneum of blade protection film, prevent that pathogenic bacteria from entering, the xylogen of plant root is increased, disease is alleviated;
2. the ester composition that contains in the complex microorganism preparations does not decompose in herbvore pest body, can make insect produce physiological barrier and death;
3. microbial strains can also form collaborative mutually mycoderm in plant materials or root, blade face field planting, can improve the dietetic alimentation ability of plant root, effectively suppresses growth and breeding and the invasion of harmful bacterium.
4. by the activity of microorganism, promote the decomposition of the soil organism and synthesizing of biologically active substance, can improve soil water-retaining, fertilizer-preserving ability, improved ecological environment of soil, suppressed the breeding of weeds in field and harmful microbe, promoted the leaf vegetables growth.
5. replenish essential trace element is provided, avoid plant in default of necessary trace element, endogenous illnesss such as generation is withered, macula lutea, leaflet, disease-controlling effect is consolidated in the combination of inner and outside.
Embodiment
The invention will be further described below in conjunction with embodiment.
Embodiment 1
A kind of microbial preparation that can reduce leaf vegetable plant diseases and insect pests is made by following raw material in weight part: beauveria bassiana nutrient solution 20, streptomycete nutrient solution 5, composite bacteria nutrient solution 30, garlic 1, molasses 40, herbal medicine 4 and micro-inorganic salt solution 0.8;
Wherein, described composite bacteria is made up of bacillus subtilis bacteria culture fluid 45wt%, Rhodopseudomonas palustris 30wt% and yeast 25wt%.
Described herbal medicine is made up of Radix Glycyrrhizae 30wt%, Japanese Honeysuckle 40wt% and Root of Indigowoad 30wt%.
Described micro-inorganic salt solution is the inorganic salt solution that contains iron ion, molybdenum ion, cupric ion, boron ion and chlorion, above-mentioned ionic concn and be 1g/L.
The above-mentioned preparation method that can reduce the microbial preparation of leaf vegetable plant diseases and insect pests, concrete steps are:
The first step: first liquid nutrient medium, is mixed with weight ratio with tween-80 after 30 minutes 120 ℃ of sterilizations at 100: 0.5; The weight ratio of beauveria bassiana by 1: 80 is inoculated in institute's liquid nutrient medium mixture of winning, at 25 ℃, oscillation frequency be under the condition of 130r/min in shaking table shaking culture 5d, sampling is observed, when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, stop fermentation, obtain the beauveria bassiana nutrient solution;
Described first liquid nutrient medium is made up of following raw material in weight part: analysis for soybean powder 4, peanut powder 2, sucrose 5, potassium primary phosphate 2, sal epsom 0.2, calcium chloride 0.02, manganous sulfate 0.005, vitamins B 8 and sterilized water 80;
Second the step: with second liquid nutrient medium 120 ℃ the sterilization 30 minutes after, the weight ratio of streptomycete by 1: 50 is inoculated in above-mentioned second liquid nutrient medium, regulating p H value is 7.0, in 25 ℃ of temperature, oscillation frequency be under the condition of 200r/min in shaking table shaking culture 8d, sampling is observed, when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, stop fermentation, obtain the streptomycete nutrient solution;
Described second liquid nutrient medium is made up of following raw material in weight part: starch 15, Semen Maydis powder 8, yeast powder 5, soybean cake powder 2, lime carbonate 1.5, potassium primary phosphate 1.5, sal epsom 0.4, calcium chloride 0.01, cobalt chloride 0.0004 and sterilized water 68;
The 3rd step: subtilis, Rhodopseudomonas palustris, yeast are mixed by 45: 30: 25 weight ratio, be inoculated in the 3rd liquid nutrient medium by 1: 50 weight ratio, the pH value is adjusted to 6.8, after 26 ℃ of speed with 150r/min stir 25min, airtight standing for fermentation 3d, sampling is observed, and stops fermentation when microscopically viable bacteria total concn number surpasses 1,000,000,000/m1, obtains the composite bacteria nutrient solution;
Described the 3rd liquid nutrient medium is made up of following raw material in weight part: molasses 10, peptone 0.5, yeast powder 1, ammonium sulfate 0.2, sal epsom 0.2 and sterilized water 88;
The 4th step: garlic and herbal medicine are pulverized the back and beauveria bassiana nutrient solution, streptomycete nutrient solution, composite bacteria nutrient solution and molasses are mixed in proportion, insert the inherent 35 ℃ of fermentation 6d of closed tin, sampling is observed, treat that viable bacteria total concn number stops fermentation when surpassing 1,200,000,000/ml, obtain the mixed fermentation product;
The 5th goes on foot: go on foot in the mixed fermentation product that obtains the 4th and add micro-inorganic salt solution in proportion, obtain reducing the microbial preparation of leaf vegetable plant diseases and insect pests.
Embodiment 2
A kind of microbial preparation that can reduce leaf vegetable plant diseases and insect pests is made by following raw material in weight part: beauveria bassiana nutrient solution 15, streptomycete nutrient solution 10, composite bacteria nutrient solution 28, garlic 11, molasses 35, herbal medicine 1 and micro-inorganic salt solution 0.2;
Wherein, described composite bacteria is made up of bacillus subtilis bacteria culture fluid 35wt%, Rhodopseudomonas palustris 40wt% and yeast 25wt%.
Described herbal medicine is made up of Radix Glycyrrhizae 50wt%, Japanese Honeysuckle 20wt% and Root of Indigowoad 30wt%.
Described micro-inorganic salt solution is the inorganic salt solution that contains iron ion, molybdenum ion, cupric ion, boron ion and chlorion, above-mentioned ionic concn and be 5g/L.
The above-mentioned preparation method that can reduce the microbial preparation of leaf vegetable plant diseases and insect pests, concrete steps are:
The first step: first liquid nutrient medium, is mixed with weight ratio with tween-80 after 30 minutes 125 ℃ of sterilizations at 100: 0.6; The weight ratio of beauveria bassiana by 1: 90 is inoculated in institute's liquid nutrient medium mixture of winning, at 28 ℃, oscillation frequency be under the condition of 120r/min in shaking table shaking culture 6d, sampling is observed, when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, stop fermentation, obtain the beauveria bassiana nutrient solution;
Described first liquid nutrient medium is made up of following raw material in weight part: analysis for soybean powder 3, peanut powder 4, sucrose 8, potassium primary phosphate 1.5, sal epsom 0.5, calcium chloride 0.01, manganous sulfate 0.006, vitamins B 9 and sterilized water 75;
Second the step: with second liquid nutrient medium 125 ℃ the sterilization 20 minutes after, the weight ratio of streptomycete by 1: 60 is inoculated in above-mentioned second liquid nutrient medium, regulating the pH value is 7.0, in 27 ℃ of temperature, oscillation frequency be under the condition of 220r/min in shaking table shaking culture 9d, sampling is observed, when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, stop fermentation, obtain the streptomycete nutrient solution;
Described second liquid nutrient medium is made up of following raw material in weight part: starch 12, Semen Maydis powder 9, yeast powder 4, soybean cake powder 4, lime carbonate 1.5, potassium primary phosphate 2, sal epsom 0.5, calcium chloride 0.01, cobalt chloride 0.0004 and sterilized water 68;
The 3rd step: subtilis, Rhodopseudomonas palustris, yeast are mixed by 35: 40: 25 weight ratio, be inoculated in the 3rd liquid nutrient medium by 1: 80 weight ratio, the pH value is adjusted to 6.8, after 28 ℃ of speed with 200r/min stir 20min, airtight standing for fermentation 4d, sampling is observed, and stops fermentation when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, obtains the composite bacteria nutrient solution;
Described the 3rd liquid nutrient medium is made up of following raw material in weight part: molasses 8, peptone 0.8, yeast powder 1, ammonium sulfate 0.5, sal epsom 0.2 and sterilized water 90;
The 4th step: garlic and herbal medicine are pulverized the back and beauveria bassiana nutrient solution, streptomycete nutrient solution, composite bacteria nutrient solution and molasses are mixed in proportion, insert the inherent 32 ℃ of fermentation 7d of closed tin, sampling is observed, treat that viable bacteria total concn number stops fermentation when surpassing 1,200,000,000/ml, obtain the mixed fermentation product;
The 5th goes on foot: go on foot in the mixed fermentation product that obtains the 4th and add micro-inorganic salt solution in proportion, obtain reducing the microbial preparation of leaf vegetable plant diseases and insect pests.
Embodiment 3
A kind of microbial preparation that can reduce leaf vegetable plant diseases and insect pests is made by following raw material in weight part: beauveria bassiana nutrient solution 20, streptomycete nutrient solution 10, composite bacteria nutrient solution 30, garlic 10, molasses 20, herbal medicine 10 and micro-inorganic salt solution 0.7;
Wherein, described composite bacteria is made up of bacillus subtilis bacteria culture fluid 25wt%, Rhodopseudomonas palustris 50wt% and yeast 25wt%.
Described herbal medicine is made up of Radix Glycyrrhizae 40wt%, Japanese Honeysuckle 40wt% and Root of Indigowoad 20wt%.
Described micro-inorganic salt solution is the inorganic salt solution that contains iron ion, molybdenum ion, cupric ion, boron ion and chlorion, above-mentioned ionic concn and 2g/L.
The above-mentioned preparation method that can reduce the microbial preparation of leaf vegetable plant diseases and insect pests, concrete steps are:
The first step: first liquid nutrient medium, is mixed with weight ratio with tween-80 after 15 minutes 130 ℃ of sterilizations at 100: 0.8; The weight ratio of beauveria bassiana by 1: 100 is inoculated in institute's liquid nutrient medium mixture of winning, at 35 ℃, oscillation frequency be under the condition of 110r/min in shaking table shaking culture 7d, sampling is observed, when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, stop fermentation, obtain the beauveria bassiana nutrient solution;
Described first liquid nutrient medium is made up of following raw material in weight part: analysis for soybean powder 2, peanut powder 5, sucrose 10, potassium primary phosphate 1.5, sal epsom 0.6, calcium chloride 0.02, manganous sulfate 0.008, vitaminB10 and sterilized water 72;
Second the step: with second liquid nutrient medium 130 ℃ the sterilization 15 minutes after, the weight ratio of streptomycete by 1: 100 is inoculated in above-mentioned second liquid nutrient medium, regulating the pH value is 7.2, in 28 ℃ of temperature, oscillation frequency be under the condition of 240r/min in shaking table shaking culture 10d, sampling is observed, when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, stop fermentation, obtain the streptomycete nutrient solution;
Described second liquid nutrient medium is made up of following raw material in weight part: starch 10, Semen Maydis powder 10, yeast powder 2, soybean cake powder 5, lime carbonate 1.5, potassium primary phosphate 2, sal epsom 0.6, calcium chloride 0.02, cobalt chloride 0.0005 and sterilized water 78.3795;
The 3rd step: subtilis, Rhodopseudomonas palustris, yeast are mixed by 25: 50: 25 weight ratio, be inoculated in the 3rd liquid nutrient medium by 1: 100 weight ratio, the pH value is adjusted to 6.8, after 35 ℃ of speed with 300r/min stir 15min, airtight standing for fermentation 4d, sampling is observed, and stops fermentation when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, obtains the composite bacteria nutrient solution;
Described the 3rd liquid nutrient medium is made up of following raw material in weight part: molasses 5, peptone 1, yeast powder 0.5, ammonium sulfate 0.5, sal epsom 0.5 and sterilized water 92.5;
The 4th step: garlic and herbal medicine are pulverized the back and beauveria bassiana nutrient solution, streptomycete nutrient solution, composite bacteria nutrient solution and molasses are mixed in proportion, insert the inherent 30 ℃ of fermentation 8d of closed tin, sampling is observed, treat that viable bacteria total concn number stops fermentation when surpassing 1,200,000,000/ml, obtain the mixed fermentation product;
The 5th goes on foot: go on foot in the mixed fermentation product that obtains the 4th and add micro-inorganic salt solution in proportion, obtain reducing the microbial preparation of leaf vegetable plant diseases and insect pests.
Embodiment 4
A kind of microbial preparation that can reduce leaf vegetable plant diseases and insect pests is made by following raw material in weight part: beauveria bassiana nutrient solution 10, streptomycete nutrient solution 10, composite bacteria nutrient solution 15, garlic 15, molasses 35, herbal medicine 15 and micro-inorganic salt solution 0.5;
Wherein, described composite bacteria is made up of bacillus subtilis bacteria culture fluid 25wt%, Rhodopseudomonas palustris 30wt% and yeast 45wt%.
Described herbal medicine is made up of Radix Glycyrrhizae 40wt%, Japanese Honeysuckle 40wt% and Root of Indigowoad 20wt%.
Described micro-inorganic salt solution is the inorganic salt solution that contains iron ion, molybdenum ion, cupric ion, boron ion and chlorion, above-mentioned ionic concn and be 3g/L.
The above-mentioned preparation method that can reduce the microbial preparation of leaf vegetable plant diseases and insect pests, concrete steps are:
The first step: first liquid nutrient medium, is mixed with weight ratio with tween-80 after 15 minutes 130 ℃ of sterilizations at 100: 0.8; The weight ratio of beauveria bassiana by 1: 100 is inoculated in institute's liquid nutrient medium mixture of winning, at 35 ℃, oscillation frequency be under the condition of 110r/min in shaking table shaking culture 7d, sampling is observed, when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, stop fermentation, obtain the beauveria bassiana nutrient solution;
Described first liquid nutrient medium is made up of following raw material in weight part: analysis for soybean powder 2, peanut powder 5, sucrose 10, potassium primary phosphate 1.5, sal epsom 0.6, calcium chloride 0.02, manganous sulfate 0.001, vitaminB10 and sterilized water 72;
Second the step: with second liquid nutrient medium 130 ℃ the sterilization 15 minutes after, the weight ratio of streptomycete by 1: 100 is inoculated in above-mentioned second liquid nutrient medium, regulating the pH value is 7.2, in 30 ℃ of temperature, oscillation frequency be under the condition of 240r/min in shaking table shaking culture 10d, sampling is observed, when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, stop fermentation, obtain the streptomycete nutrient solution;
Described second liquid nutrient medium is made up of following raw material in weight part: starch 10, Semen Maydis powder 10, yeast powder 2, soybean cake powder 5, lime carbonate 1.5, potassium primary phosphate 2, sal epsom 0.6, calcium chloride 0.02, cobalt chloride 0.0007 and sterilized water 80;
The 3rd step: subtilis, Rhodopseudomonas palustris, yeast are mixed by 25: 30: 45 weight ratio, be inoculated in the 3rd liquid nutrient medium by 1: 100 weight ratio, the pH value is adjusted to 6.8, after 35 ℃ of speed with 300r/min stir 15min, airtight standing for fermentation 2d, sampling is observed, and stops fermentation when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, obtains the composite bacteria nutrient solution;
Described the 3rd liquid nutrient medium is made up of following raw material in weight part: molasses 5, peptone 1, yeast powder 0.5, ammonium sulfate 0.5, sal epsom 0.5 and sterilized water 94;
The 4th step: garlic and herbal medicine are pulverized the back and beauveria bassiana nutrient solution, streptomycete nutrient solution, composite bacteria nutrient solution and molasses are mixed in proportion, insert the inherent 30 ℃ of fermentation 8d of closed tin, sampling is observed, treat that viable bacteria total concn number stops fermentation when surpassing 1,200,000,000/ml, obtain the mixed fermentation product;
The 5th goes on foot: go on foot in the mixed fermentation product that obtains the 4th and add micro-inorganic salt solution in proportion, obtain reducing the microbial preparation of leaf vegetable plant diseases and insect pests.
The present invention can take different usages and different ratios during use according to demand:
Foliage-spray: dilution 200-300 doubly carries out vaporific sprinkling to plant stem-leaf portion, and the blade tow sides all will be sprayed onto; Take the circumstances into consideration to reduce extension rate in the more serious plot of preceding stubble disease and pest.
Pouring water reclamation: water after diluting 1000-1500 times;
Soil improvement: when the leaf vegetables bulk application, evenly admix use, every mu of general 2-3kg that uses; Use when crop growth period, should apply after loosening the soil, every mu of 5-6kg waters after the use.
Practical application effect of the present invention is as follows:
This product began to have carried out pakchoi plantation contrast test in Songjiang, Shanghai five village, often used in village names agricultural garden the first tenday period of a month in May, 2009, The using method that experimental group adopts foliage-spray to combine with soil improvement. When executing base manure, pakchoi evenly admixes use, and every Mu uses 2-3kg; After the pakchoi seed transplanted for 1 week, use 200-300 doubly to dilute liquid plant stem-leaf section carried out vaporific sprinkling, After this sprayed once every 7-10 days. Experimental result shows that the present invention can make the pakchoi disease and pest reduce 40-45%, Xiao Bai Dish output increased 10-15%, ripe 7-10 days in advance of pakchoi.
Soil is detected: the ratio of Fusarium≤5% in the thread bacterium of soil; Total nitrogen content improves 5% in the soil; Effectively Phosphorus content improves more than 8%; Content of organics improves 10%.
Economic benefit: agricultural chemicals, Reducing amount of chemical fertilizer applied are more than 30%, and indirect cost reduces more than 25%.
Claims (5)
1, a kind of microbial preparation that can reduce leaf vegetable plant diseases and insect pests, it is characterized in that it is made by following raw material in weight part: beauveria bassiana nutrient solution 10-20, streptomycete nutrient solution 5-10, composite bacteria nutrient solution 15-30, garlic 1-15, molasses 20-40, herbal medicine 1-15 and micro-inorganic salt solution 0.2-0.8;
Wherein, described composite bacteria is made up of bacillus subtilis bacteria culture fluid 25-45wt%, Rhodopseudomonas palustris 30-50wt% and yeast 25-45wt%.
2, the microbial preparation that can reduce leaf vegetable plant diseases and insect pests as claimed in claim 1 is characterized in that, described herbal medicine is made up of Radix Glycyrrhizae 40wt%, Japanese Honeysuckle 40wt% and Root of Indigowoad 20wt%.
3, the microbial preparation that can reduce leaf vegetable plant diseases and insect pests as claimed in claim 1, it is characterized in that, described micro-inorganic salt solution is the inorganic salt solution that contains iron ion, molybdenum ion, cupric ion, boron ion and chlorion, above-mentioned ionic concn and be 1-5g/L.
4, the described preparation method that can reduce the microbial preparation of leaf vegetable plant diseases and insect pests of claim 1 is characterized in that concrete steps are:
The first step: with first liquid nutrient medium 120-130 ℃ of sterilization after 15-30 minute, with tween-80 with weight ratio 100: 0.5-0.8 mixes; With beauveria bassiana by 1: the weight ratio of 80-100 be inoculated in the institute liquid nutrient medium mixture of winning in, at 25-35 ℃, oscillation frequency be under the condition of 110-130r/min in shaking table shaking culture 5-7d, sampling is observed, when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, stop fermentation, obtain the beauveria bassiana nutrient solution;
Described first liquid nutrient medium is made up of following raw material in weight part: analysis for soybean powder 2-4, peanut powder 2-5, sucrose 5-10, potassium primary phosphate 1.5-2, sal epsom 0.2-0.6, calcium chloride 0.01-0.02, manganous sulfate 0.001-0.008, VB8-10 and sterilized water 70-80;
Second the step: with second liquid nutrient medium 120-130 ℃ the sterilization 15-30 minute after, with streptomycete by 1: the weight ratio of 50-100 is inoculated in above-mentioned second liquid nutrient medium, regulating p H value is 7.0-7.2, at temperature 25-30 ℃, oscillation frequency be under the condition of 200-240r/min in shaking table shaking culture 8-10d, sampling is observed, when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, stop fermentation, obtain the streptomycete nutrient solution;
Described second liquid nutrient medium is made up of following raw material in weight part: starch 10-15, Semen Maydis powder 8-10, yeast powder 2-5, soybean cake powder 2-5, lime carbonate 1.5-2.0, potassium primary phosphate 1.5-2, sal epsom 0.2-0.6, calcium chloride 0.01-0.02, cobalt chloride 0.0004-0.0007 and sterilized water 65-78;
The 3rd step: subtilis, Rhodopseudomonas palustris, yeast are pressed 25-45: 30-50: the weight ratio of 25-45 is mixed, by 1: the weight ratio of 50-100 is inoculated in the 3rd liquid nutrient medium, the pH value is adjusted to 6.8, after the 25-35 ℃ of speed with 150-300r/min stirs 15-25min, airtight standing for fermentation 2-4d, sampling is observed, and stops fermentation when microscopically viable bacteria total concn number surpasses 1,000,000,000/ml, obtains the composite bacteria nutrient solution;
Described the 3rd liquid nutrient medium is made up of following raw material in weight part: molasses 5-10, peptone 0.5-1, yeast powder 0.5-1, ammonium sulfate 0.2-0.5, sal epsom 0.2-0.5 and sterilized water 88-94;
The 4th step: garlic and herbal medicine are pulverized the back and beauveria bassiana nutrient solution, streptomycete nutrient solution, composite bacteria nutrient solution and molasses are mixed in proportion, insert the inherent 30-35 ℃ of fermentation 6-8d of closed tin, sampling is observed, treat that viable bacteria total concn number stops fermentation when surpassing 1,200,000,000/ml, obtain the mixed fermentation product;
The 5th goes on foot: go on foot in the mixed fermentation product that obtains the 4th and add micro-inorganic salt solution in proportion, obtain reducing the microbial preparation of leaf vegetable plant diseases and insect pests.
5, the preparation method that can reduce the microbial preparation of leaf vegetable plant diseases and insect pests as claimed in claim 4 is characterized in that, the temperature of shaking culture is 26-28 ℃ in described second step.
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