CN112341284A - Special microbial fertilizer for tobacco and preparation method and application thereof - Google Patents
Special microbial fertilizer for tobacco and preparation method and application thereof Download PDFInfo
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- CN112341284A CN112341284A CN202011328846.XA CN202011328846A CN112341284A CN 112341284 A CN112341284 A CN 112341284A CN 202011328846 A CN202011328846 A CN 202011328846A CN 112341284 A CN112341284 A CN 112341284A
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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
- C05G3/60—Biocides or preservatives, e.g. disinfectants, pesticides or herbicides; Pest repellants or attractants
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05B—PHOSPHATIC FERTILISERS
- C05B1/00—Superphosphates, i.e. fertilisers produced by reacting rock or bone phosphates with sulfuric or phosphoric acid in such amounts and concentrations as to yield solid products directly
- C05B1/02—Superphosphates
Abstract
The invention discloses a tobacco microbial fertilizer, which comprises four strains of Saccharomyces cerevisiae (Saccharomyces cerevisiae), Streptomyces lavandulae (Streptomyces lavendaae), Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) and Trichoderma viride (Trichoderma viride). The microbial agent provided by the invention utilizes the synergistic effect of the four bacteria to quickly form a dominant flora in the tobacco microbial fertilizer, is compatible with the raw materials such as sophora alopecuroides straw and the like, fully decomposes the raw materials and promotes the fertilizer efficiency to be exerted. Meanwhile, the problem that the microbial fertilizer cannot give consideration to both fertilizer efficiency and disease and insect resistance is solved. The prepared microbial fertilizer can remarkably promote the growth of tobacco, has remarkable control effects on tobacco bacterial wilt, tobacco black shank, tobacco mosaic disease, root knot nematode disease and myzus persicae, and has good ecological benefit and economic benefit and huge development potential.
Description
Technical Field
The invention belongs to the technical field of biological fermentation, and further relates to a tobacco fertilizer, and further relates to the technical field of a special microbial fertilizer for tobacco and application thereof.
Background
The biological bacterial fertilizer usually takes organic or inorganic fertilizer as a carrier, and the growth of plants is regulated by adding a certain special microorganism and utilizing the combined action of the microorganism and the common fertilizer. The biological bacterial fertilizer mainly has two functions, namely, the biological bacterial fertilizer is used as a functional fertilizer; secondly, regulating the growth of plants. Research shows that after the biological bacterial manure is sprayed or applied to the tobacco leaf surfaces, the plant root systems grow vigorously, the root system activity is improved, and the nutrient absorption capacity is enhanced. The bacterial manure can increase the chlorophyll content of tobacco plants, improve the photosynthetic rate of tobacco leaves, increase the leaf area, promote the yield increase of the tobacco and improve the quality of the tobacco leaves. Research also finds that the disease resistance of the tobacco is improved after the biological bacterial manure is applied, but the comprehensive influence of the biological bacterial manure on tobacco cultivation is rarely reported.
Currently, common diseases and pests of tobacco are as follows: the prevention and control of brown spot, black shank, bacterial wilt, mosaic disease, root knot nematode disease and myzus persicae mainly adopt chemical prevention and control, agricultural prevention and control, biological prevention and control and other measures, but the characteristic of difficult prevention and control dominates the strategy after the scale generation. The tobacco leaves produced by only applying the organic fertilizer have good quality but low yield. The tobacco leaves produced by simply applying inorganic fertilizer have greatly improved yield but reduced quality. Therefore, the fertilizer applied to the tobacco needs to have long fertilizer efficiency so as to avoid the damage of the root system caused by frequent fertilization, and the nutrient needs to be supplied stably so as to ensure the uniformity of the leaves and improve the quality of the tobacco leaves. The biological bacterial fertilizer is an environment-friendly fertilizer, is one of important directions of future agricultural sustainable development, and has the advantages of maintaining and improving soil fertility, being beneficial to propagation of dominant species of microorganisms, effectively preventing and controlling soil-borne diseases, keeping ecological balance, reducing environmental pollution and the like.
The special biological bacterial fertilizer for grass provided by the invention patent with the application number of 201810692750.8 in the prior art can effectively prevent and control black shank, bacterial wilt and root knot nematode, but is not ideal for preventing and controlling myzus persicae and mosaic disease.
Disclosure of Invention
Based on the incomplete prevention and control of plant diseases and insect pests by the special bacterial fertilizer for tobacco prepared in the prior art, the invention aims to provide the special bacterial fertilizer which has the obvious effects of simultaneously preventing and controlling tobacco black shank, bacterial wilt, mosaic disease, root knot nematode disease and myzus persicae and can obviously promote the growth of tobacco. The microbial agent provided by the invention utilizes the synergistic effect of the four bacteria to quickly form a dominant flora in the tobacco microbial fertilizer, the raw materials are compatible, the raw materials are fully decomposed, and the fertilizer efficiency is promoted to be exerted. Meanwhile, the problem that the microbial fertilizer cannot give consideration to both fertilizer efficiency and disease and insect resistance is solved. The prepared microbial fertilizer can remarkably promote the growth of tobacco, has remarkable prevention and treatment effects on various plant diseases and insect pests, has good ecological and economic benefits and has huge development potential.
The present invention solves the above-mentioned problems by the following technical means. Specifically, the special tobacco microbial fertilizer comprises, by weight, 10-18 parts of sophora alopecuroides straw, 4-12 parts of biomass charcoal, 31-39 parts of calcium superphosphate, 21-29 parts of potassium sulfate, 10-14 parts of ammonium nitrate and 4-8 parts of a microbial agent.
Preferably, the microbial fertilizer special for tobacco comprises, by weight, 14 parts of sophora alopecuroide straw, 8 parts of biomass charcoal, 35 parts of calcium superphosphate, 25 parts of potassium sulfate, 12 parts of ammonium nitrate and 6 parts of a microbial agent.
The special tobacco microbial fertilizer is prepared by providing biomass charcoal prepared by mixing and cleaning rice husk charcoal and rice straw charcoal according to the mass ratio of 1: 1.
The special microbial fertilizer for the tobacco is prepared by four strains of Saccharomyces cerevisiae (Saccharomyces cerevisiae), Streptomyces lavandulae (Streptomyces lavendale), Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) and Trichoderma viride (Trichoderma viride) according to a specific ratio and a preparation method thereof, and is further applied to tobacco planting, so that the raw materials can be fully decomposed, the fertilizer efficiency can be promoted to be exerted, the growth of the tobacco can be obviously promoted, the obvious prevention and control effect on various plant diseases and insect pests is realized, and the special microbial fertilizer for the tobacco has good ecological benefit and economic benefit.
The invention also discloses a preparation method of the special microbial fertilizer for tobacco, which comprises the following steps:
(1) activating each slant strain stored in an environment at 0-4 ℃, and respectively carrying out test tube-shake flask culture on four strains of Saccharomyces cerevisiae (Saccharomyces cerevisiae), Streptomyces lavandulae (Streptomyces lavendaae), Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) and Trichoderma viride (Trichoderma viride) to obtain a first-grade seed solution of each strain;
(2) performing fermentation tank enlargement culture on the seed solution, fermenting Saccharomyces cerevisiae, Streptomyces lavandulae, Bacillus amyloliquefaciens and Trichoderma viride respectively, detecting light absorption value and flora number of the fermented liquid by microscopic direct counting method and ultraviolet spectrophotometry, determining growth condition of flora, and detecting effective viable bacteria number of 2 × 109Stopping culturing when cfu/mL to obtain a zymogen liquid;
(3) after the fermentation of the single strains is finished, mixing saccharomyces cerevisiae, streptomyces lavandulae, bacillus amyloliquefaciens and trichoderma viride according to the proportion of 1: (1-3): (1-5): (2-4) mixing the components in a volume ratio to obtain a composite zymocyte liquid;
(4) mixing the compound bacterial liquid obtained in the step (3) with the sophora alopecuroides straw crushed to 1-2cm, and uniformly stirring;
(5) uniformly stirring the biomass charcoal, the calcium superphosphate, the potassium sulfate and the ammonium nitrate in the raw materials obtained in the step (4), and drying until the water content is below 15 percent and the viable count is not less than 0.5 multiplied by 109cfu/mL, namely preparing the special microbial fertilizer for tobacco.
The invention provides the special microbial fertilizer for tobacco, which is obtained by the preparation process.
In the invention, a strain culture medium and a culture method adopted by Saccharomyces cerevisiae (Saccharomyces cerevisiae) are disclosed, wherein the FC2 culture medium comprises the following components in percentage by weight: 2% of sucrose, 0.4% of yeast extract, 0.3% of peptone, 0.2% of disodium hydrogen phosphate, 1-2% of inoculation amount and 150-200 rpm, and culturing for 24-36 h.
In the invention, the Streptomyces lavendensis (Streptomyces lavendensis) adopts an optimized soybean meal culture medium, and the formula of the culture medium is as follows: 1.2% of starch, 2% of glycerol, 2.1% of soybean meal, 0.6% of glucose, 0.2% of yeast extract, 0.3% of peptone, 0.2% of calcium carbonate and 2% of corn oil, wherein the inoculation amount is 10% and 220rpm, and the culture is carried out for 140 hours.
In the invention, an optimized Landy culture medium is adopted by Bacillus amyloliquefaciens (Bacillus amyloliquefaciens), and the formula of the culture medium is as follows: 2 percent of glucose, 0.5 percent of L-sodium glutamate, MgSO40.04 percent, 0.05 percent of KCl and KH2PO41％、FeSO40.15‰、MnSO45.0‰、CuSO40.14 per mill, 5 percent of inoculation amount, 160rpm, and culturing for 36 hours.
In the invention, Trichoderma viride (Trichoderma viride) is cultured for 96h by adopting a conventional PDB culture medium with the inoculum size of 5 percent and the rpm of 180.
In the invention, the adopted Saccharomyces cerevisiae (Saccharomyces cerevisiae), Streptomyces lavandulae (Streptomyces lavendale), Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) and Trichoderma viride (Trichoderma viride) are all harmless and safe strains allowed to be used by the ministry of agriculture, and are all purchased from China general microbiological culture collection center (CGMCC), and common technicians can obtain the strains through public channels.
Meanwhile, the invention provides the application of the special tobacco microbial fertilizer in agricultural wastes.
Further, the invention provides application of the special tobacco microbial fertilizer, wherein the application amount of the special tobacco microbial fertilizer is 60-140kg per mu.
Preferably, the application amount of the special tobacco microbial fertilizer is 100kg per mu.
By implementing the technical scheme of the invention, the following beneficial effects can be achieved:
(1) in the preparation process of the special microbial fertilizer for tobacco, the four strains of Saccharomyces cerevisiae (Saccharomyces cerevisiae), Streptomyces lavandulae (Streptomyces lavendaae), Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) and Trichoderma viride (Trichoderma viride) have strong adaptability synergistic effect, the microbial agent which is composed according to a specific proportion utilizes the synergistic effect of the four bacteria in the tobacco microbial fertilizer, integrates the antagonism of Streptomyces lavandulae (Streptomyces lavendale) and tobacco ralstonia solanacearum, the antagonism of Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) to tobacco mosaic virus and tobacco black shank bacteria and the antagonism of Trichoderma viride (Trichoderma viride) to other common disease viruses of tobacco, coordinates the acid production function of Saccharomyces cerevisiae (Saccharomyces cerevisiae), the pH value of the special bacterial manure can be effectively adjusted, the alkalinity is reduced, and the bacterial manure is matched according to a specific proportion, so that the integral formation of the dominant flora with disease and insect resistance and obvious growth promoting effect is promoted.
(2) According to the technical scheme provided by the invention, the microbial agent with a specific ratio is compatible with the alopecuroide straw, the rice husk carbon and the biomass carbon prepared by mixing the rice straw carbon and other raw materials, so that the insect-resistant characteristic of the alopecuroide straw and the adsorption and bearing effect of the biomass carbon on the microbial agent are fully exerted; the adopted rice husk carbon can increase potassium, various spots and even death of plant leaves caused by potassium deficiency are avoided, and the influence of potassium carbonate on acidity and alkalinity is removed through water washing; the adopted rice straw carbon can adsorb pollutants, and the mixed biomass carbon can loosen, ventilate and darken soil and promote fertilizer efficiency to play.
(3) When the special microbial fertilizer for tobacco provided by the invention is applied to tobacco planting, the activity of the bacterial strain is kept for a long time after the bacterial agent is inoculated, the dominant bacterial flora is quickly formed, the growth requirement of tobacco is ensured, the tobacco yield is obviously improved, the yield of the upper part, the middle part and the lower part and the total yield are both increased, the control effect on common diseases and insect pests of tobacco is also obvious, the control effect on tobacco bacterial wilt, tobacco black shank, tobacco mosaic disease, root knot nematode disease and tobacco aphid is obvious, and the ecological benefit and the economic benefit are good.
Drawings
FIG. 1 is a production flow chart of bacterial manure special for tobacco flower and leaf.
Detailed Description
The present invention will be described below by way of examples, but the present invention is not limited to the following examples.
The materials of the invention are: saccharomyces cerevisiae (Saccharomyces cerevisiae), Streptomyces lavendulae (Streptomyces lavendaae), Bacillus amyloliquefaciens (Bacillus amyloliquefaciens), Trichoderma viride (Trichoderma viride), bean straws, biomass charcoal, calcium superphosphate, potassium sulfate, ammonium nitrate, sucrose, yeast extract, peptone, disodium hydrogen phosphate, starch, glycerol, soybean meal, glucose, yeast extract, calcium carbonate, corn oil, glucose, sodium L-glutamate, magnesium sulfate, potassium chloride, potassium dihydrogen phosphate, ferric sulfate, manganese sulfate and copper sulfate can be purchased through public channels (sold in the market), and equipment and instruments adopted in the process are common equipment in the field.
All materials, reagents and equipment selected for use in the present invention are well known in the art, but do not limit the practice of the invention, and other reagents and equipment well known in the art may be suitable for use in the practice of the following embodiments of the invention.
The following examples further illustrate the present invention but are not to be construed as limiting the invention. Modifications or substitutions to methods, procedures, or conditions of the invention may be made without departing from the spirit and scope of the invention.
The first embodiment is as follows: special microbial fertilizer for tobacco and preparation method thereof
The special microbial fertilizer for tobacco comprises, by weight, 10-18 parts of sophora alopecuroide straw, 4-12 parts of biomass charcoal, 31-39 parts of calcium superphosphate, 21-29 parts of potassium sulfate, 10-14 parts of ammonium nitrate and 4-8 parts of a microbial agent.
The microbial agent comprises Saccharomyces cerevisiae (Saccharomyces cerevisiae), Streptomyces lavandulae (Streptomyces lavendale), Bacillus amyloliquefaciens (Bacillus amyloliquefaciens), and Trichoderma viride (Trichoderma viride).
The biomass charcoal is prepared by mixing and cleaning rice hull charcoal and rice straw charcoal according to a mass ratio of 1: 1.
The invention also discloses a preparation method of the special microbial fertilizer for tobacco, which comprises the following steps:
(1) activating various slant strains stored in an environment at 0-4 ℃, and respectively carrying out test tube-shake flask culture on four strains of Saccharomyces cerevisiae (Saccharomyces cerevisiae), Streptomyces lavandulae (Streptomyces lavendale), Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) and Trichoderma viride (Trichoderma viride), wherein the Saccharomyces cerevisiae (Saccharomyces cerevisiae) adopts an optimized FC2 culture medium, and the formula of the culture medium is as follows: 2% of sucrose, 0.4% of yeast extract, 0.3% of peptone, 0.2% of disodium hydrogen phosphate, 1-2% of inoculation amount, 150-200 rpm, and culturing for 24-36 h at 37 ℃; the lavender Streptomyces lavipeditum (Streptomyces lavendensis) adopts an optimized soybean meal culture medium, and the formula of the culture medium is as follows: 1.2% of starch, 2% of glycerol, 2.1% of soybean meal, 0.6% of glucose, 0.2% of yeast extract, 0.3% of peptone, 0.2% of calcium carbonate and 2% of corn oil, wherein the inoculation amount is 10% and 220rpm, and the culture is carried out for 140 hours at 35 ℃; an optimized Landy culture medium is adopted by Bacillus amyloliquefaciens (Bacillus amyloliquefaciens), and the formula of the culture medium is as follows: 2% of glucose, 0.5% of L-sodium glutamate, 0.04% of magnesium sulfate, 0.05% of potassium chloride, 1% of monopotassium phosphate, 0.15% of ferric sulfate, 5.0% of manganese sulfate and 0.14% of copper sulfate, 5% of inoculum size, 160rpm and 30 ℃ for 36 hours; culturing Trichoderma viride (Trichoderma viride) with conventional PDB culture medium at an inoculum size of 5% and 180rpm at 27 deg.C for 96 hr; obtaining first-stage seed liquid of each strain;
(2) performing fermentation tank enlargement culture on the seed solution, fermenting Saccharomyces cerevisiae, Streptomyces lavandulae, Bacillus amyloliquefaciens and Trichoderma viride respectively, detecting light absorption value and flora number of the fermented liquid by microscopic direct counting method and ultraviolet spectrophotometry, determining growth condition of flora, and detecting effective viable bacteria number of 2 × 109Stopping culturing when cfu/mL to obtain a zymogen liquid;
(3) after the fermentation of the single strains is finished, mixing saccharomyces cerevisiae, streptomyces lavandulae, bacillus amyloliquefaciens and trichoderma viride according to the proportion of 1: (1-3): (1-5): (2-4) mixing the components in a volume ratio to prepare a composite fermentation microbial inoculum;
(4) mixing the composite microbial inoculum liquid obtained in the step (3) with the sophora alopecuroides straw crushed to 1-2cm, and uniformly stirring;
(5) uniformly stirring the biomass charcoal, the calcium superphosphate, the potassium sulfate and the ammonium nitrate in the raw materials obtained in the step (4), and drying until the water content is below 15 percent and the viable count is not less than 0.5 multiplied by 109cfu/mL, namely preparing the special microbial fertilizer for tobacco.
Example two: preparation of special microbial fertilizer for tobacco
Activating each slant strain stored in an environment of 0-4 ℃, and respectively performing test tube-shake flask culture on four strains of saccharomyces cerevisiae, streptomyces lavandulae, bacillus amyloliquefaciens and trichoderma viride, wherein the saccharomyces cerevisiae adopts an optimized FC2 culture medium, and the formula of the culture medium is as follows: 2% of sucrose, 0.4% of yeast extract, 0.3% of peptone, 0.2% of disodium hydrogen phosphate, 1-2% of inoculation amount, 150-200 rpm, and culturing for 24-36 h at 37 ℃; the lavender streptomyces adopts an optimized soybean meal culture medium, and the formula of the culture medium is as follows: 1.2% of starch, 2% of glycerol, 2.1% of soybean meal, 0.6% of glucose, 0.2% of yeast extract, 0.3% of peptone, 0.2% of calcium carbonate and 2% of corn oil, wherein the inoculation amount is 10% and 220rpm, and the culture is carried out for 140 hours at 35 ℃; the bacillus amyloliquefaciens adopts an optimized Landy culture medium, and the formula of the culture medium is as follows: 2% of glucose, 0.5% of L-sodium glutamate, 0.04% of magnesium sulfate, 0.05% of potassium chloride, 1% of monopotassium phosphate, 0.15% of ferric sulfate, 5.0% of manganese sulfate and 0.14% of copper sulfate, 5% of inoculum size, 160rpm and 30 ℃ for 36 hours; culturing Trichoderma viride with conventional PDB culture medium at an inoculum size of 5% at 180rpm at 27 deg.C for 96 hr; obtaining first-stage seed liquid of each strain; performing fermentation tank enlargement culture on the seed solution, fermenting respectively, detecting light absorption value and flora number of the fermented liquid by microscopy direct counting method and ultraviolet spectrophotometry, determining growth condition of flora, and detecting effective viable bacteria number of 2 × 109Stopping culturing when cfu/mL to obtain a zymogen liquid; after fermentation is finished, mixing saccharomyces cerevisiae, streptomyces lavandulae, bacillus amyloliquefaciens and trichoderma viride according to the proportion of 1: 1: 1: 2, preparing a composite fermentation microbial inoculum; mixing 4kg of the composite microbial inoculum liquid with 10kg of sophora alopecuroides straw crushed to 1-2cm, and uniformly stirring; adding rice husk carbon and rice straw carbon into the raw materials according to the mass ratio of 1:1Mixing and cleaning to obtain biomass charcoal 4kg, calcium superphosphate 31kg, potassium sulfate 21kg and ammonium nitrate 10kg, stirring, drying to water content below 15%, and viable count not less than 0.5 × 109cfu/mL, namely preparing the special microbial fertilizer for tobacco. 60kg of the fertilizer is applied to each mu.
Example three: preparation of special microbial fertilizer for tobacco
Activating and culturing each bacterium based on the second embodiment to obtain a zymogen liquid; after fermentation is finished, mixing saccharomyces cerevisiae, streptomyces lavandulae, bacillus amyloliquefaciens and trichoderma viride according to the proportion of 1: 1: 2: 2, preparing a composite fermentation microbial inoculum; 5kg of composite microbial inoculum liquid is mixed with 12kg of sophora alopecuroides straw which is crushed to 1-2cm and is stirred uniformly; adding rice hull carbon and rice straw carbon into the raw materials, mixing and cleaning according to the mass ratio of 1:1 to obtain 6kg of biomass carbon, 33kg of calcium superphosphate, 23kg of potassium sulfate and 11kg of ammonium nitrate, uniformly stirring, drying until the water content is below 15%, and the viable count is not less than 0.5 multiplied by 109cfu/mL, namely preparing the special microbial fertilizer for tobacco. 80kg of the fertilizer is applied to each mu.
Example four: preparation of special microbial fertilizer for tobacco
Activating and culturing each bacterium based on the second embodiment to obtain a zymogen liquid; after fermentation is finished, mixing saccharomyces cerevisiae, streptomyces lavandulae, bacillus amyloliquefaciens and trichoderma viride according to the proportion of 1: 2: 3: 3, mixing the components in a volume ratio to prepare a composite fermentation microbial inoculum; mixing 6kg of the composite microbial inoculum liquid with 14kg of sophora alopecuroides straw crushed to 1-2cm, and uniformly stirring; adding rice hull carbon and rice straw carbon into the raw materials, mixing and cleaning according to the mass ratio of 1:1 to obtain 8kg of biomass carbon, 35kg of calcium superphosphate, 25kg of potassium sulfate and 12kg of ammonium nitrate, uniformly stirring, drying until the water content is below 15%, and the viable count is not less than 0.5 multiplied by 109cfu/mL, namely preparing the special microbial fertilizer for tobacco. 100kg is applied to each mu when the fertilizer is applied.
Example five: preparation of special microbial fertilizer for tobacco
Activating and culturing each bacterium based on the second embodiment to obtain a zymogen liquid; after fermentation is finished, mixing saccharomyces cerevisiae, streptomyces lavandulae, bacillus amyloliquefaciens and trichoderma viride according to the proportion of 1: 2: 4: 3 volume ratio ofMixing to obtain a compound fermentation microbial inoculum; mixing 7kg of the composite microbial inoculum liquid with 16kg of sophora alopecuroides straw crushed to 1-2cm, and uniformly stirring; adding rice hull carbon and rice straw carbon into the raw materials, mixing and cleaning according to the mass ratio of 1:1 to obtain 10kg of biomass carbon, 37kg of calcium superphosphate, 27kg of potassium sulfate and 13kg of ammonium nitrate, uniformly stirring, drying until the water content is below 15%, and the viable count is not less than 0.5 multiplied by 109cfu/mL, namely preparing the special microbial fertilizer for tobacco. When in application, 120kg of the fertilizer is applied to each mu.
Example six: preparation of special microbial fertilizer for tobacco
Activating and culturing each bacterium based on the second embodiment to obtain a zymogen liquid; after fermentation is finished, mixing saccharomyces cerevisiae, streptomyces lavandulae, bacillus amyloliquefaciens and trichoderma viride according to the proportion of 1: 3: 5: 4, preparing the composite fermentation inoculum; mixing 8kg of the composite microbial inoculum liquid with 18kg of sophora alopecuroides straw crushed to 1-2cm, and uniformly stirring; adding rice hull carbon and rice straw carbon into the raw materials, mixing and cleaning according to the mass ratio of 1:1 to obtain 12kg of biomass carbon, 39kg of calcium superphosphate, 29kg of potassium sulfate and 14kg of ammonium nitrate, uniformly stirring, drying until the water content is below 15%, and the viable count is not less than 0.5 multiplied by 109cfu/mL, namely preparing the special microbial fertilizer for tobacco. 140kg of the fertilizer is applied to each mu.
Example seven: optimization of preparation process of special microbial fertilizer for tobacco
The following different formulas are respectively adopted to prepare the components of the complex microbial inoculum:
formula 1: the method comprises the following steps of 1: 1: 1: 2, and mixing to obtain the composite microbial inoculum.
And (2) formula: the method comprises the following steps of 1: 1: 2: 2, and mixing to obtain the composite microbial inoculum.
And (3) formula: the method comprises the following steps of 1: 2: 3: 3, and mixing to obtain the composite microbial inoculum.
And (4) formula: the method comprises the following steps of 1: 2: 4: 3, and mixing to obtain the composite microbial inoculum.
And (5) formula: the method comprises the following steps of 1: 3: 5: 4, and mixing to obtain the composite microbial inoculum.
The five different formulas are respectively prepared into the complex microbial inoculum according to the following processes:
activating each slant strain stored in an environment of 0-4 ℃, and respectively performing test tube-shake flask culture on four strains of saccharomyces cerevisiae, streptomyces lavandulae, bacillus amyloliquefaciens and trichoderma viride, wherein the saccharomyces cerevisiae adopts an optimized FC2 culture medium, and the formula of the culture medium is as follows: 2% of sucrose, 0.4% of yeast extract, 0.3% of peptone, 0.2% of disodium hydrogen phosphate, 1-2% of inoculation amount, 150-200 rpm, and culturing for 24-36 h at 37 ℃; the lavender streptomyces adopts an optimized soybean meal culture medium, and the formula of the culture medium is as follows: 1.2% of starch, 2% of glycerol, 2.1% of soybean meal, 0.6% of glucose, 0.2% of yeast extract, 0.3% of peptone, 0.2% of calcium carbonate and 2% of corn oil, wherein the inoculation amount is 10% and 220rpm, and the culture is carried out for 140 hours at 35 ℃; the bacillus amyloliquefaciens adopts an optimized Landy culture medium, and the formula of the culture medium is as follows: 2% of glucose, 0.5% of L-sodium glutamate, 0.04% of magnesium sulfate, 0.05% of potassium chloride, 1% of monopotassium phosphate, 0.15% of ferric sulfate, 5.0% of manganese sulfate and 0.14% of copper sulfate, 5% of inoculum size, 160rpm and 30 ℃ for 36 hours; culturing Trichoderma viride with conventional PDB culture medium at an inoculum size of 5% at 180rpm at 27 deg.C for 96 hr; obtaining first-stage seed liquid of each strain; performing fermentation tank enlargement culture on the seed solution, fermenting respectively, detecting light absorption value and flora number of the fermented liquid by microscopy direct counting method and ultraviolet spectrophotometry, determining growth condition of flora, and detecting effective viable bacteria number of 2 × 109Stopping culturing when cfu/mL to obtain a zymogen liquid; after fermentation is finished, mixing saccharomyces cerevisiae, streptomyces lavandulae, bacillus amyloliquefaciens and trichoderma viride according to a volume ratio to prepare a composite fermentation microbial inoculum; mixing 6kg of the composite microbial inoculum liquid with 14kg of sophora alopecuroides straw crushed to 1-2cm, and uniformly stirring; adding rice hull carbon and rice straw carbon into the raw materials, mixing and cleaning according to the mass ratio of 1:1 to obtain 8kg of biomass carbon35kg of calcium phosphate, 25kg of potassium sulfate and 12kg of ammonium nitrate, uniformly stirring, drying until the water content is below 15 percent and the viable count is not less than 0.5 multiplied by 109cfu/mL, namely preparing the special microbial fertilizer for tobacco. 100kg is applied to each mu when the fertilizer is applied.
1. Orthogonal optimization composite microbial inoculum
A single-factor experiment is designed, the influence of the fertilizer efficiency of the special tobacco microorganism bacteria prepared by the streptomyces lavandulae, the bacillus amyloliquefaciens and the trichoderma viride in different adding proportions is respectively explored, and the influence on the total tobacco yield is detected. Three-factor three-level orthogonal experiments were performed on the basis of the single-factor experiments, see table 1.
Table 1: orthogonality test factors and levels
The results and analysis of the orthogonal optimization test are shown in Table 2.
Table 2: results and analysis of orthogonal assays
As can be seen from the table 2, in the range of the experimental design, the influence of the bacillus amyloliquefaciens and the trichoderma viride on the fertilizing effect of the compound microbial inoculum is the largest, and the influence of the streptomyces lavandulae is smaller. The optimal raw material of the complex microbial inoculum is prepared as A2B2C2Namely, streptomyces lavandulae, bacillus amyloliquefaciens and trichoderma viride are mixed according to the volume ratio of 2: 3: 3, and preparing the special microbial fertilizer for tobacco according to the preparation process provided by the fourth embodiment.
Example eight: effect of applying special microbial fertilizer for tobacco in tobacco planting
1. Bacterial manure preparation
The microbial fertilizer special for tobacco is prepared according to the second embodiment to the sixth embodiment.
2. And (3) field test:
summary of the test: the test soil has moderate fertility, smooth tobacco land, yellow soil, irrigation condition and better illumination condition. The test plots are plots with serious diseases of perennial bacterial wilt, black shank, mosaic disease, root knot nematode disease and myzus persicae.
3. And (3) field test design:
the microbial fertilizer special for tobacco prepared according to the second to sixth embodiments is applied according to the application amount defined in each embodiment, and is respectively numbered 1-5, the microbial fertilizer prepared by the technical scheme provided in the invention patent with the application number of 201810692750.8 is used as a reference 1 group, a conventional fertilizer is used as a reference 2 group, the microbial fertilizer special for tobacco prepared by removing the straw component of sophora alopecuroides based on the technical scheme provided in the fourth embodiment is used as a reference 3 group, the microbial fertilizer prepared by removing the biomass charcoal component based on the technical scheme provided in the fourth embodiment is used as a reference 4 group, the test is totally 3 rows, and the application mode is repeated for 3 times: hole application of base fertilizer and sprinkling application of additional fertilizer.
3. Analysis items and methods:
(1) investigation of disease onset
The occurrence of black shank, bacterial wilt, mosaic disease, root knot nematode disease and myzus persicae is investigated every 30 days from 30 days after fertilization, and the investigation is continuously carried out for 3 times. And the average incidence was calculated. The calculation method is as follows:
and counting the number of the myzus persicae and the root-knot nematodes in each square meter of soil. The results are shown in Table 3.
Table 3: disease and pest prevention effect of applying special microbial fertilizer for tobacco to tobacco planting
Through determination, the special microbial fertilizer for tobacco of each group prepared by the technical scheme provided by the invention has a better effect, and compared with the microbial fertilizer prepared by the technical scheme provided by the invention patent with the application number of 201810692750.8 as a comparison group 1 and the conventional chemical fertilizer as a comparison group 2, the special microbial fertilizer has obvious effects of reducing the occurrence conditions of black shank, bacterial wilt, mosaic disease, root knot nematode disease and myzus persicae, and is respectively reduced by 0%, 60.7% and 72% compared with the comparison group 1; compared with a control 2 group, the reduction is respectively 26.7%, 30%, 82%, 71.7% and 84.4%; the control 3 groups have control effects on black shank and bacterial wilt which are lower than those of the control 1 group and higher than those of the control 2 group, have control effects on mosaic diseases which are better than those of the control 1 group and the control 2 group but are obviously lower than those of the test 1-5 groups, and have control effects on myzus persicae which are lower than those of the other groups; the control effect of the control group 4 on diseases is lower than that of the control groups 1-5, 2 and 3, and the control effect on insect pests is similar to that of the control groups 1-5. Therefore, the addition of the sophora alopecuroides straw can obviously contribute to the effect of the special bacterial fertilizer for tobacco on pest control in the tobacco planting process, and meanwhile, the addition of the sophora alopecuroides straw in different proportions can also have different degrees of influence on the pest control effect of the special bacterial fertilizer for tobacco. Moreover, different microbial inoculum proportions provided by the invention have a significant effect on pest control, different degrees of dominant floras are formed by different proportions, so that different effects on pest control are achieved, and the addition of biomass charcoal has an effect on the adsorption effect of the microbial inoculum, so that the disease control is affected.
(2) Method for measuring yield and quality of tobacco leaves
And (3) taking the cell as a unit, separately harvesting and baking, and respectively calculating the yield of the upper leaves, the yield of the middle leaves, the yield of the lower leaves and the total yield, and finally converting the yield per mu. The results are shown in Table 4.
Table 4: growth promoting effect of special microbial fertilizer for tobacco in tobacco planting
Through determination, the special microbial fertilizer for tobacco of each group prepared by the technical scheme provided by the invention has a better effect, and compared with the microbial fertilizer prepared by the technical scheme provided by the invention patent with the application number of 201810692750.8 as a comparison group 1 and the conventional fertilizer applied as a comparison group 2, the special microbial fertilizer has obvious effects of improving the levels of upper leaf yield, middle leaf yield, lower leaf yield and total yield by 10.44%, 2.90%, 2.22% and 4.38% respectively compared with the comparison group 1; compared with a control group 2, the content of the protein is respectively increased by 30.68%, 21.82%, 29.12% and 25.67%; the levels of the comparison group 3 on the upper leaf yield, the middle leaf yield, the lower leaf yield and the total yield are higher than those of the comparison group 1 and the comparison group 2, but lower than those of the experiments 1-5, so that the addition of the sophora alopecuroides straw not only can influence the control effect of the special bacterial fertilizer for the tobacco on plant diseases and insect pests, but also indirectly influences each part and the total yield of the tobacco. Meanwhile, by comparing the effects of the test groups 1-5, the effect of adding the microbial inoculum with different strain proportions on the pest control can be found out, and the tobacco yield is influenced to different degrees. The yield of the control group 4 is lower than that of the test groups 1-5, the control group 1 and the control group 3, but higher than that of the control group 2, so that the addition of the biomass charcoal has an influence on the adsorption effect of the microbial inoculum so as to influence the overall yield of the tobacco. The optimal technical effect obtained by the technical scheme provided by the fourth embodiment is better than that of other groups in the effect of promoting the growth of tobacco.
In conclusion, the microbial agent is prepared by mixing Saccharomyces cerevisiae (Saccharomyces cerevisiae), Streptomyces lavandulae (Streptomyces lavendale), Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) and Trichoderma viride (Trichoderma viride) according to a specific proportion, and the microbial agent is compounded with biomass carbon and other components which are prepared by mixing sophora alopecuroides straw, rice hull carbon and rice straw carbon according to a specific proportion.
As described above, the present invention can be preferably implemented, and the above-mentioned embodiments only describe the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various changes and modifications of the technical solution of the present invention made by those skilled in the art without departing from the design spirit of the present invention shall fall within the protection scope defined by the present invention.
Claims (10)
1. The special microbial fertilizer for tobacco is characterized by comprising, by weight, 10-18 parts of sophora alopecuroides straw, 4-12 parts of biomass charcoal, 31-39 parts of calcium superphosphate, 21-29 parts of potassium sulfate, 10-14 parts of ammonium nitrate and 4-8 parts of a microbial agent.
2. The microbial fertilizer special for tobacco according to claim 1, comprising 14 parts by weight of sophora alopecuroides straw, 8 parts by weight of biomass charcoal, 35 parts by weight of calcium superphosphate, 25 parts by weight of potassium sulfate, 12 parts by weight of ammonium nitrate and 6 parts by weight of microbial agent.
3. The microbial fertilizer special for tobacco according to any one of claims 1 or 2, characterized in that the microbial agent comprises Saccharomyces cerevisiae (Saccharomyces cerevisiae), Streptomyces lavandulae (Streptomyces lavendaae), Bacillus amyloliquefaciens (Bacillus amyloliquefaciens), and Trichoderma viride (Trichoderma viride).
4. The microbial fertilizer special for tobacco according to any one of claims 1 or 2, wherein the biomass charcoal is prepared by mixing rice husk charcoal and rice straw charcoal according to a mass ratio of 1: 1.
5. The method for preparing the special microbial fertilizer for tobacco as claimed in claim 1, which comprises the following steps:
(1) activating each slant strain stored in an environment at 0-4 ℃, and respectively carrying out test tube-shake flask culture on four strains of Saccharomyces cerevisiae (Saccharomyces cerevisiae), Streptomyces lavandulae (Streptomyces lavendaae), Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) and Trichoderma viride (Trichoderma viride) to obtain a first-grade seed solution of each strain;
(2) performing fermentation tank enlargement culture on the seed solution, fermenting Saccharomyces cerevisiae, Streptomyces lavandulae, Bacillus amyloliquefaciens and Trichoderma viride, respectively, detecting light absorption value and flora number of the fermented liquid by microscopic direct counting method and ultraviolet spectrophotometry, determining growth condition of flora, and detecting effective viable bacteria number of 2 × 109Stopping culturing at cfu/mL to obtain each single-strain zymogen liquid;
(3) after the fermentation of the single strains is finished, fermenting bacteria liquid of saccharomyces cerevisiae, streptomyces lavandulae, bacillus amyloliquefaciens and trichoderma viride is mixed according to the proportion of 1: (1-3): (1-5): (2-4) mixing in a volume ratio to obtain a composite zymocyte liquid;
(4) mixing the compound bacterial liquid obtained in the step (3) with the sophora alopecuroides straw crushed to 1-2cm, and uniformly stirring;
(5) adding biomass charcoal, calcium superphosphate, potassium sulfate and ammonium nitrate into the raw material obtained in the step (4), uniformly stirring, and drying until the water content is below 15 percent and the viable count is not less than 0.5 multiplied by 109cfu/mL, namely preparing the special microbial fertilizer for tobacco.
6. The method for preparing the microbial fertilizer special for tobacco as claimed in claim 5, wherein saccharomyces cerevisiae adopts an optimized FC2 culture medium, and the formula of the culture medium is as follows: 2% of sucrose, 0.4% of yeast extract, 0.3% of peptone, 0.2% of disodium hydrogen phosphate, 1-2% of inoculation amount and 150-200 rpm, and culturing for 24-36 h.
7. The method for preparing the microbial fertilizer special for tobacco as claimed in claim 5, wherein the streptomyces lavandulae adopts an optimized soybean meal culture medium, and the formula of the culture medium is as follows: 1.2% of starch, 2% of glycerol, 2.1% of soybean meal, 0.6% of glucose, 0.2% of yeast extract, 0.3% of peptone, 0.2% of calcium carbonate and 2% of corn oil, wherein the inoculation amount is 10% and 220rpm, and the culture is carried out for 140 hours.
8. The method for preparing the microbial fertilizer special for tobacco as claimed in claim 5, wherein the bacillus amyloliquefaciens adopts an optimized Landy culture medium, and the formula of the culture medium is as follows: 2% of glucose, 0.5% of L-sodium glutamate, 0.04% of magnesium sulfate, 0.05% of potassium chloride, 1% of monopotassium phosphate, 0.15% of ferric sulfate, 5.0% of manganese sulfate and 0.14% of copper sulfate, 5% of inoculum size, 160rpm and culturing for 36 h.
9. The method for preparing the microbial fertilizer special for tobacco as claimed in claim 5, wherein the Trichoderma viride is cultured for 96h by using a conventional PDB culture medium with an inoculum size of 5% and at 180 rpm.
10. The method for applying the microbial fertilizer special for tobacco as claimed in any one of claims 1 to 9, wherein the application amount of the microbial fertilizer special for tobacco is 60-140kg per mu.
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| CN117511753A (en) * | 2023-11-23 | 2024-02-06 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | Marine trichoderma strain and application thereof in prevention and treatment of tobacco mosaic virus |
| CN117511753B (en) * | 2023-11-23 | 2024-04-26 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | Marine trichoderma strain and application thereof in prevention and treatment of tobacco mosaic virus |
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