CN112794746B

CN112794746B - Composite bacterial powder for Chinese yam and preparation method thereof, and composite bacterial fertilizer for Chinese yam and preparation method thereof

Info

Publication number: CN112794746B
Application number: CN202110288165.3A
Authority: CN
Inventors: 焦国宝; 孙宁宁; 高璇璇; 朱丽娜; 陈晓宇; 崔红健; 王渝鹭; 曹忠洋; 张永钦; 朱飞帆; 段波; 刘芦鹏; 卢佳琦; 邹瑞瑞; 李佳; 谢莹娟; 席华强
Original assignee: Henan New Yangshao Biological Technology Co ltd
Current assignee: Henan New Yangshao Biological Technology Co ltd
Priority date: 2021-03-16
Filing date: 2021-03-16
Publication date: 2022-09-02
Anticipated expiration: 2041-03-16
Also published as: CN112794746A

Abstract

The invention provides composite bacterial powder for Chinese yam and a preparation method thereof, and composite bacterial fertilizer for Chinese yam and a preparation method thereof, and relates to the technical field of Chinese yam planting. The composite bacterial powder for the Chinese yam comprises the following composite bacterial powder according to the viable count ratio: the composite fungus powder is (20-100): (1-10), wherein the composite fungus powder comprises bacillus subtilis, bacillus laterosporus, bacillus polymyxa and bacillus thuringiensis, and the composite fungus powder comprises trichoderma harzianum and metarhizium anisopliae. The compound bacterial fertilizer for the Chinese yam comprises, by mass, 200-500 parts of compound bacterial powder for the Chinese yam, 200-500 parts of fulvic acid potassium, 100-300 parts of algal polysaccharide and 5-30 parts of chitosan oligosaccharide. Bacteria and fungi in the compound bacterial fertilizer for Chinese yam work cooperatively to improve soil structure, prevent diseases and pests and promote plant growth; the bacterial manure keeps physiological active substances secreted by microorganisms, has high contents of nutrition and active ingredients, and improves the yield of the Chinese yam through comprehensive action.

Description

Compound bacterial powder for Chinese yam and preparation method thereof, and compound bacterial fertilizer for Chinese yam and preparation method thereof

Technical Field

The invention relates to the technical field of Chinese yam planting, in particular to composite bacterial powder for Chinese yam and a preparation method thereof, and composite bacterial fertilizer for Chinese yam and a preparation method thereof.

Background

The yam has dual purposes of medicine and food, the root tuber is fat and deep, not only has more nutrition needed, but also is easy to be attacked by soil insects such as cutworms, grubs, wireworms and the like, so that the yam has reduced yield and deteriorated quality. The overground vine of the Chinese yam is dense, the environment is concealed, a high-temperature and high-humidity environment is created, and the invasion of pathogenic bacteria is facilitated.

The following defects exist in the current yam planting:

firstly, the yield is increased by applying a large amount of chemical fertilizer;

secondly, frequently using pesticides to prevent plant diseases and insect pests;

thirdly, the available microbial products have single strain, low activity and single effect.

In view of the above, the present invention is particularly proposed.

Disclosure of Invention

The invention aims to provide composite bacterial powder for Chinese yam, which solves the technical problems of single strain, low activity and single effect of the prior art that the yield is increased by applying a large amount of chemical fertilizer, pesticides are frequently used for preventing diseases and insect pests, and available microbial products are frequently used.

The invention also aims to provide a preparation method of the composite bacteria powder for the yam, which has the advantages of low cost, maintenance of metabolites of each microorganism and high content of nutrients and active ingredients.

The invention also aims to provide the compound bacterial fertilizer for the Chinese yam, which can stimulate the growth of plants according to the growth and development characteristics of the Chinese yam, provide nutritional ingredients for the Chinese yam in all aspects, improve soil conditions and prevent diseases and insect pests of the Chinese yam.

The invention also aims to provide a preparation method of the composite bacterial fertilizer for the yam, which is simple and easy to operate and is suitable for large-scale industrial production.

In order to solve the technical problems, the invention adopts the following technical scheme:

the invention provides composite bacterial powder for Chinese yam in a first aspect, which comprises the following components in viable count ratio: compound bacterial powder: the composite fungus powder is (20-100) to (1-10);

wherein the composite bacterial powder comprises bacillus subtilis, bacillus laterosporus, bacillus polymyxa and bacillus thuringiensis; the composite fungus powder comprises trichoderma harzianum and metarhizium anisopliae.

Furthermore, the viable bacteria ratio of the bacillus subtilis, the bacillus laterosporus, the bacillus polymyxa and the bacillus thuringiensis in the composite bacterial powder is (50-200): 1-10: (1-10): 10-30);

and/or the viable count ratio of trichoderma harzianum to metarhizium anisopliae in the composite fungus powder is (1-10) to (5-20).

Furthermore, the viable count ratio of the bacillus subtilis, the bacillus laterosporus, the bacillus polymyxa and the bacillus thuringiensis in the composite bacterial powder is (100-150): 2-8): 20-28;

and/or the viable count ratio of trichoderma harzianum to metarhizium anisopliae in the composite fungus powder is (2-8) to (10-15).

The second aspect of the present invention provides the preparation method of the composite bacterial powder for yam of the first aspect, wherein the composite bacterial powder for yam is obtained by mixing the composite bacterial powder and the composite fungal powder.

Further, the preparation method of the composite bacterial powder comprises the following steps:

step a: respectively activating the strains of the bacillus subtilis, the bacillus laterosporus, the bacillus polymyxa and the bacillus thuringiensis to respectively obtain four activated bacteria liquids;

step b: b, respectively preparing four seed solutions from the four activated bacterial solutions obtained in the step a;

step c: c, respectively carrying out liquid fermentation culture on the four seed liquids obtained in the step b to obtain corresponding four fermentation liquids;

step d: c, mixing the four fermentation liquors obtained in the step c, and then carrying out spray drying;

preferably, the speed of the spray drying is 800-1300 ml/h;

preferably, the spray drying is carried out in a spray drying tower, wherein the inlet temperature of the spray drying tower is 145-150 ℃, and the outlet temperature of the spray drying tower is 90-95 ℃.

Further, the preparation method of the composite fungus powder comprises the following steps:

step A: respectively activating the trichoderma harzianum and metarhizium anisopliae to respectively obtain two activated strains;

and B: b, respectively carrying out liquid fermentation culture on the two activated strains obtained in the step A to obtain two corresponding mother solutions;

and C: respectively inoculating the two mother solutions obtained in the step B to a solid culture medium;

step D: drying the two solid culture media obtained in the step C at a low temperature and mixing to obtain the composite fungus powder;

wherein in the step D, the temperature of the low-temperature drying is not more than 35 ℃.

The third aspect of the invention provides a compound bacterial fertilizer for Chinese yam, which comprises compound bacterial powder for Chinese yam, a carrier, a fertilizer synergist and a biological stimulator;

the composite bacterial powder for Chinese yam comprises the composite bacterial powder for Chinese yam in the first aspect or the composite bacterial powder for Chinese yam prepared by the preparation method in the second aspect.

Further, the carrier comprises potassium fulvate;

and/or, the fertilizer synergist comprises algal polysaccharides;

and/or, the biostimulant comprises a chitosan oligosaccharide.

Further, the composition comprises the following components in parts by mass: 200-500 parts of composite bacterial powder for Chinese yam, 200-500 parts of potassium fulvate, 100-300 parts of algal polysaccharide and 5-30 parts of chitosan oligosaccharide.

The fourth aspect of the invention provides a preparation method of the compound bacterial fertilizer for the Chinese yam of the third aspect, which is obtained by uniformly mixing the compound bacterial powder for the Chinese yam, the potassium fulvic acid, the algal polysaccharides and the chitosan oligosaccharide.

According to the composite bacterial powder for the Chinese yam, provided by the invention, the characteristics of growth and pest and disease damage of the Chinese yam are combined, bacillus subtilis, bacillus laterosporus, bacillus polymyxa, bacillus thuringiensis, trichoderma harzianum and metarhizium anisopliae are selected for compounding, the soil structure is improved through cooperative work, meanwhile, diseases and pests are prevented, and the growth of plants is promoted. When the floras are used in combination, the inhibition efficiency of the plant diseases and insect pests of the Chinese yam is higher than the sum of single floras, and the Chinese yam is non-toxic and pollution-free, and the Chinese yam plants do not generate drug resistance.

The preparation method of the compound bacterial powder for the Chinese yam, provided by the invention, has the advantages that the seed liquid after the activation of the strains is fermented, the preparation cost is low, the physiological active substances secreted by microorganisms, such as cytokinin, indoleacetic acid, gibberellin, vitamins, amino acid and the like, are reserved, and the contents of nutrients and active ingredients are high. The substances promote new root germination, plant growth and tuber expansion, and indirectly improve the yield of the Chinese yam.

According to the compound bacterial fertilizer for the Chinese yam, the microbial agent forms a protective barrier through modes of competition, antibiotic, bacteriolysis and the like, so that the growth and propagation of pathogenic bacteria are inhibited; the carrier, the fertilizer synergist and the biological stimulator provide nutrient substances for plant growth, improve the stress resistance of plants, chelate metal ions and are beneficial to plant utilization. The components of the compound bacterial fertilizer have synergistic effect, and can prevent and treat plant diseases, enhance plant resistance, promote plant growth and improve yam yield.

The preparation method of the compound bacterial fertilizer for the Chinese yam provided by the invention is obtained by uniformly mixing the compound bacterial powder for the Chinese yam, the potassium fulvic acid, the algal polysaccharide and the chitosan oligosaccharide, the proportion can be adjusted according to the characteristics of the cultivation soil, and the compound bacterial powder for the raw materials can still keep the activity of the strains. Simple process, labor saving and suitability for large-scale industrial production.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention clearer, the technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Thus, the following detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Due to the specific planting and growing mode of the Chinese yam, the growth and the quality of the Chinese yam can be influenced greatly by soil conditions. Considering the characteristic of deep root pricking of the Chinese yam, part of the Chinese yam can even be pricked to the soil below 1m, and the pest control must be enhanced in the growth process of the Chinese yam. Common diseases of Chinese yam include anthracnose, brown spot, root-stem rot and blight, and are mainly caused by colletotrichum gloeosporioides, yam alternaria, fusarium solani or fusarium oxysporum in soil. Common underground pests of yam include cutworms, wireworms and grubs. The invention provides a comprehensive prevention and control method aiming at the plant diseases and insect pests.

According to the first aspect of the invention, the composite bacterial powder for the Chinese yam comprises the following components in viable count ratio: compound bacterial powder: the composite fungus powder is (20-100) to (1-10);

According to the composite bacterial powder for the Chinese yam, provided by the invention, the characteristics of growth and pest and disease damage of the Chinese yam are combined, bacillus subtilis, bacillus laterosporus, bacillus polymyxa, bacillus thuringiensis, trichoderma harzianum and metarhizium anisopliae are selected for compounding, the soil structure is improved through cooperative work, and the problem that the yield is increased by applying a large amount of chemical fertilizers in the prior art is solved. Meanwhile, the pesticide composition can prevent diseases and insect pests, promote the growth of plants and reduce the use amount of the existing pesticide. When the floras are prepared according to a certain proportion, the inhibition efficiency of the floras on the plant diseases and insect pests of the Chinese yam is higher than the sum of single bacteria, the technical problems of single strain, low activity and single effect of the existing microbial products are solved, and the Chinese yam plant is non-toxic and pollution-free and does not generate drug resistance.

Bacillus subtilis is a species of Bacillus, CAS number 68038-70-0. The single cell is 0.7-0.8 multiplied by 2-3 microns and is uniformly colored. Without capsule, the perigenic flagellum can move. Gram-positive bacteria can form endogenous adversity resistant spores, the spores are 0.6-0.9 multiplied by 1.0-1.5 microns, the ellipse to the column are positioned in the center of the bacteria or slightly deviated, and the bacteria do not expand after the spores are formed. The growth and propagation speed is high, the surface of a colony is rough and opaque, and is white or yellowish, and when the colony grows in a liquid culture medium, the skin becomes always formed, so that the colony is an aerobic bacterium. Active substances such as subtilin, polymyxin, nystatin, gramicidin and the like generated in the growth process of bacillus subtilis have obvious inhibiting effect on pathogenic bacteria or conditional pathogenic bacteria of endogenous infection. Bacillus subtilis also produces antibiotics which inhibit the growth of bacteria, viruses, fungi and pathogens, and these antibiotics are based on phospholipids, aminosugars, peptides and lipopeptides. Among them, lipopeptide antibiotics are the most important antibacterial substances of bacillus subtilis.

The bacillus laterosporus improves the soil micro-ecological environment, promotes the formation of a granular structure, improves the soil texture, increases the number of beneficial microorganisms in the soil, increases the ratio of gram-positive bacteria to gram-negative bacteria and improves the soil micro-ecological environment. The mass propagation of the bacillus laterosporus generates viscous secretion, increases soil organic matters, promotes the formation of a soil aggregate structure, and improves the permeability, the soil moisture retention, the heat preservation and the fertilizer preservation of soil.

The bacillus polymyxa exists in a spore form, has good temperature tolerance and stable quality, treats bacteria with bacteria and has lasting drug effect; the pesticide and fertilizer can prevent and control soil-borne diseases such as bacterial wilt, fusarium wilt, root rot, soft rot and the like, and has the obvious effects of promoting growth, increasing the yield of Chinese yam and improving the quality of Chinese yam.

The bacillus thuringiensis strain can produce endotoxin and exotoxin to stop pests from eating, and the pests die due to hunger, cell wall rupture, blood destruction and nerve poisoning. After the bacillus thuringiensis is eaten by pests, the bacillus thuringiensis parasitizes in the midgut of a host and grows and propagates in a proper alkaline environment in the intestine, and crystal toxins are hydrolyzed by protease in the intestinal tract of a pest body to form smaller subunits with toxic effects, and the subunits act on epithelial cells of the midgut of the pest body to cause intestinal paralysis, perforation, paralysis of the pest body and stop eating. The bacillus thuringiensis then enters the blood cavity to multiply, causing septicemia, resulting in death of the insect.

Trichoderma harzianum has preventing effect on plant diseases caused by pathogenic bacteria such as pythium, Rhizoctonia solani, fusarium, Rhizopus nigricans, Cylindrocarpon, Sclerotinia sclerotiorum and the like in the growth environment of Chinese yam. The trichoderma harzianum grows rapidly at the root and leaf peripheries of the plants, occupies the sites on the surface of the plants, forms a protective cover, prevents pathogenic bacteria from contacting the root systems and the surfaces of the leaves of the plants, thereby protecting the roots and the leaves of the plants from being infected by the pathogenic bacteria and ensuring that the plants can grow healthily.

The metarhizium anisopliae forms a rhizosphere symbiotic relationship at the root of the Chinese yam to be subjected to long-term storage, pests in soil actively invade through the body wall of the pests to infect different pests, yeast-shaped propagation can be performed in the blood cavity of the pests to generate a large amount of worm bodies, meanwhile, insecticidal toxins such as destructor and the like can be generated, the immunity of the pests is inhibited, and the pests are killed quickly. When the pest host does not exist, the metarhizium anisopliae can form rhizosphere symbiotic relationship with the root of the plant to carry out long-term storage, and long-term protection is provided for the growth of the Chinese yam plant.

The ratio of bacteria to fungi affects the growth of yam plants and yam yield. The ratio of bacteria to fungi is set in the range of (20-100) to (1-10). In a preferred embodiment of the invention, the ratio of the viable count of the composite bacterial powder to the composite fungal powder in the composite bacterial powder for Chinese yam is typically, but not limited to, 100:1, 50:1, 20:1, 10:1, 2: 1.

Furthermore, the viable bacteria ratio of the bacillus subtilis, the bacillus laterosporus, the bacillus polymyxa and the bacillus thuringiensis in the composite bacterial powder is (50-200): (1-10): (10-30).

The ratio of viable bacteria of Bacillus subtilis, Bacillus laterosporus, Bacillus polymyxa and Bacillus thuringiensis in the composite bacterial powder is typically, but not limited to, 50:1:1:10, 20:1:1:3, 10:1:1:1 or 200:1:1: 10.

The ratio of viable bacteria of the composite fungal powder is typically, but not limited to, 2:1, 1:2, 1:5, 1:10 or 1: 20.

Furthermore, the viable bacteria ratio of the bacillus subtilis, the bacillus laterosporus, the bacillus polymyxa and the bacillus thuringiensis in the composite bacterial powder is (100-150): (2-8): (20-28).

In a preferred embodiment of the invention, the viable count ratio of Bacillus subtilis, Bacillus laterosporus, Bacillus polymyxa and Bacillus thuringiensis is typically, but not limited to, 50:1:1:10, 25:2:2:5, 30:2:2:7 or 30:2:2: 5.

In a preferred embodiment of the invention, the viable count ratio of Trichoderma harzianum to Metarhizium anisopliae is typically, but not limited to, 1:5, 2:15, 4:5, 8:15 or 1: 3.

According to the preparation method of the composite bacterial powder for the Chinese yam provided by the second aspect of the invention, the composite bacterial powder for the Chinese yam is obtained by mixing the composite bacterial powder and the composite fungal powder.

According to the preparation method of the compound bacterial powder for the Chinese yam, provided by the invention, the seed liquid after the strain activation is fermented, the preparation cost is low, the metabolites of the microorganisms are reserved, and the contents of nutrients and active ingredients are high.

a, step a: respectively activating the strains of the bacillus subtilis, the bacillus laterosporus, the bacillus polymyxa and the bacillus thuringiensis to respectively obtain four activated bacteria liquids;

preferably, the speed of the spray drying is 800-1300 ml/h;

The strain activation is to put the preserved strain into a proper culture medium for culture, and to enlarge the culture step by step to obtain a pure and strong culture, i.e. to obtain a culture with vigorous activity and enough inoculation quantity. The strain fermentation has a rejuvenation process which generally needs 2-3 generations, and the condition during storage is different from the condition during culture, so the strain needs to be activated and gradually adapts to the culture environment.

The activated strains are subjected to expanding culture, the fermentation period is shortened, and the bacteria are cultured by fermentation as early as possible after coming to a stable period. Fermentation culture is the process of mass propagation of bacteria under suitable growth conditions. The liquid culture medium is used in the bacterial culture process, nutrient substances of the liquid culture medium are uniformly distributed and fully contacted with the surfaces of thalli, and metabolic products of microorganisms can be greatly dissolved. Therefore, a large amount of bacteria and bacterial metabolites, such as cytokinin, indoleacetic acid, gibberellin, vitamins, amino acids, etc., are present in the fermentation broth of the fermentation culture.

Spray drying is a method for applying systematic technology to material drying. After the mixed fermentation liquor is atomized in a drying chamber, water is quickly vaporized in the contact with hot air to obtain dried bacterial powder, so that the damage and the killing to the bacteria in the crushing process are avoided.

In the spray drying process, potassium fulvate is diluted according to the viable bacteria concentration of the target bacterial powder.

The spray drying is carried out in a spray drying tower at a rate typically, but not limited to, 800ml/h, 1000ml/h or 1300 ml/h. The inlet temperature of the fermentation broth in the spray drying tower is typically, but not limited to, 145 ℃, 146 ℃, 147 ℃, 148 ℃, 149 ℃ or 150 ℃, and the outlet temperature of the bacterial powder in the spray drying tower is typically, but not limited to, 90 ℃, 91 ℃, 92 ℃, 93 ℃, 94 ℃ or 95 ℃.

step A: respectively activating the trichoderma harzianum and the metarhizium anisopliae to respectively obtain two activated strains;

step D: drying the two solid culture mediums obtained in the step C at a low temperature and mixing to obtain the composite fungus powder;

According to a third aspect of the invention, the compound bacterial fertilizer for the Chinese yam comprises compound bacterial powder for the Chinese yam, a carrier, a fertilizer synergist and a biological stimulator;

The compound bacterial fertilizer for the Chinese yam provided by the invention can comprehensively guarantee the growth of Chinese yam plants by adding bacterial powder, a carrier, a fertilizer synergist and a biological stimulator, wherein various thalli are arranged to be beneficial to soil remediation and soil improvement, and can play a role in inhibiting bacteria, resisting insects and diseases, prevent soil hardening and acidification, enhance soil permeability, and the thalli fermentation of the preparation method adopts a compound fermentation process, so that the compound bacterial fertilizer is rich in bioactive substances such as extracellular polysaccharide and the like, so that the crops are high in yield and quality, and the soil can be continuously and efficiently cultivated. The carrier, the fertilizer synergist and the biological stimulator act together to enhance the stress resistance of cold resistance, drought resistance and the like of crops and the capability of resisting plant diseases and insect pests.

The carrier is a main substance for bearing the bacterial powder, and after the carrier is applied to soil, the carrier can provide nutrients such as organic matters for microorganisms, is beneficial to growth and propagation, and prevents the low content of the organic matters in the soil and the limited growth and propagation of the microorganisms.

The fertilizer synergist can directly act on crops and can also improve the soil fertilizer efficiency. Mineral nutrition which is difficult to utilize in the soil can be activated, and medium and trace elements which are necessary for crops can be supplemented; the harmful substances in the soil can be removed, the soil is purified and repaired, and the soil structure is improved; can promote the activity of microorganisms, enhance the stress resistance of crops and improve the quality.

The target of the biological stimulant is the Chinese yam plant, which can improve the physiological and biochemical state of the plant and improve the pesticide effect and the utilization rate of the fertilizer. Activating the innate immune system of the plant, rooting, inhibiting bacteria, preventing diseases, resisting stress and improving quality.

Further, the carrier comprises potassium fulvate;

and/or, the fertilizer synergist comprises algal polysaccharides;

and/or, the biostimulant comprises a chitosan oligosaccharide.

In a preferred embodiment of the invention, the carrier is typically, but not limited to, potassium fulvate. The potassium fulvate is beneficial to the growth of Chinese yam plants, can chelate metal ions and is beneficial to the utilization of the plants; preventing and controlling plant diseases and enhancing resistance; improve nutrient absorption and promote plant growth.

In a preferred embodiment of the invention, the fertilizer synergist is typically, but not limited to, algal polysaccharides. The algal polysaccharide provides nutrient substances for the growth and development of microorganisms, and is beneficial to the growth and the propagation of the microorganisms. Can stimulate the growth of Chinese yam plants and improve the stress resistance of the plants, such as cold resistance, disease resistance, waterlogging resistance and the like.

In a preferred embodiment of the invention, the biostimulant is typically, but not limited to, chitosan oligosaccharide. The chitosan oligosaccharide changes the soil flora and promotes the growth of beneficial microorganisms. The chitosan oligosaccharide can also induce disease resistance of Chinese yam plants, and has immunity and killing effects on various fungi, bacteria, viruses and nematodes. The compound algal polysaccharide promotes the absorption of nutrient elements of Chinese yam plants, promotes the growth of root systems, increases the yield, strengthens the plants, enhances the stress resistance, achieves the effects of deep and luxuriant roots and leaves, improves the quality and improves the bacteriostatic and antibacterial effects of the Chinese yam.

In some preferred embodiments of the present invention, the yam composite bacterial powder is typically, but not limited to, 200 parts, 300 parts, 400 parts or 500 parts; potassium fulvate is typically, but not limited to, 200 parts, 300 parts, 400 parts or 500 parts; algal polysaccharides are typically, but not limited to, 100 parts, 200 parts or 300 parts; chitosan oligosaccharides are typically, but not limited to, 5 parts, 10 parts, 20 parts or 30 parts.

According to the preparation method of the compound bacterial fertilizer for the Chinese yam provided by the fourth aspect of the invention, the compound bacterial powder for the Chinese yam, the potassium fulvic acid, the algal polysaccharides and the chitosan oligosaccharides are uniformly mixed to obtain the compound bacterial fertilizer.

The preparation method of the compound bacterial fertilizer for the Chinese yam, provided by the invention, is simple in process, labor-saving and suitable for large-scale industrial production. The composite bacterial fertilizer for the Chinese yam fully utilizes the interaction principle of plants and microorganisms with soil and the active response principle of root systems to the environment, takes rhizosphere microorganisms and fertilizer nutrients as entry points, regulates the air permeability of the soil through organic matters on the basis of meeting the nitrogen, phosphorus and potassium nutrients required by the Chinese yam, promotes the effective absorption of calcium through the chelation of potassium fulvate to calcium ions, has the function of regulating the growth of the Chinese yam, stimulates the extending growth of the root systems of the Chinese yam and the generation of fibrous roots, and finally achieves the effects of promoting the growth and development of the Chinese yam plants and improving the yield.

The following examples are provided to explain the embodiments of the present invention in detail. The embodiments described below and the features of the embodiments can be combined with each other without conflict.

Example 1

The embodiment provides a compound bacterial powder for Chinese yam, which comprises the following specific steps:

step 1: respectively inoculating the preserved strains of the bacillus subtilis, the bacillus laterosporus, the bacillus polymyxa and the bacillus thuringiensis on an LB plate culture medium, culturing for 40h at 36 ℃, transferring the strains to a shake flask of an LB liquid culture medium for further culture, wherein the culture temperature is 36 ℃, the rotation speed of a shaking table is 145r/min, and the culture time is 36 h. Respectively transferring the activated bacteria liquid into respective seed tanks, wherein the inoculation amount is 3%, the culture temperature is 36 ℃, the pressure is 0.8MPa, and the ventilation capacity is 50m ³ And h, fermenting for 24h to obtain a seed solution. Respectively transferring the prepared seed liquid into respective liquid fermentation culture medium for fermentation culture.

B, fermentation culture of the bacillus subtilis: the inoculation amount is 5-8%, the culture temperature is 32-36 ℃, the rotation speed is 180- ³ And h, stopping fermentation when the spore rate reaches 98%. The culture medium is composed of 8-10 parts of corn flour, 25-30 parts of soybean meal, 3-5 parts of sodium chloride and 3-5 parts of ammonium sulfate, and the raw materials of the culture medium are crushed by a superfine crusher. The pH value is 6.5-7.0.

B, fermentation culture of bacillus laterosporus: the inoculation amount is 5-8%, the culture temperature is 32-35 ℃, and the rotation speed is 160-; the ventilation capacity is 60-80m ³ And h, stopping fermentation when the spore rate reaches 98%. The culture medium consists of corn flour 15-20 weight portions, soybean cake 10-15 weight portions, sodium dihydrogen phosphate 1-2 weight portions and magnesium sulfate 0.5-0.8 weight portion, and the material is superfine crushedAnd (4) crushing by a machine. The pH value is 6.0-7.0.

And (3) fermenting and culturing the bacillus polymyxa: the inoculation amount is 8-10%, the culture temperature is 32-35 ℃, the rotating speed is 120-150r/min, and the ventilation volume is 50-80m ³ And h, stopping fermentation when the spore rate reaches 98 percent. The culture medium is composed of 15-20 parts of corn starch, 30-35 parts of soybean meal, 4-7 parts of sodium chloride and 3-5 parts of calcium carbonate, and the raw materials of the culture medium are crushed by a superfine crusher. The pH value is 6.0-7.2.

And (3) fermenting and culturing the bacillus thuringiensis: the inoculation amount is 8-13%, the culture temperature is 28-35 ℃, the rotation speed is 160- ³ And h, stopping fermentation when the spore rate reaches 98 percent. The culture medium is composed of 10-15 parts of corn starch, 10-15 parts of soybean meal, 3-6 parts of ammonium sulfate, 2-4 parts of dipotassium hydrogen phosphate, 0.4-0.8 part of magnesium sulfate, 0.4-0.6 part of sodium chloride and 0.05-0.1 part of ferrous sulfate, and the raw materials of the culture medium are crushed by a superfine crusher. The pH value is 6.5-7.5.

And 2, step: and (2) mixing the bacillus subtilis, bacillus laterosporus, bacillus polymyxa and bacillus thuringiensis fermentation liquor obtained in the step (1) according to the viable count ratio of 50:1:1:10, and then performing spray drying by using a spray drying tower, wherein the feeding speed is 1000ml/h, the inlet temperature is 145 ℃, and the outlet temperature is 95 ℃ to obtain bacterial powder.

And step 3: respectively inoculating the preserved Trichoderma harzianum and Metarhizium anisopliae to a PDA plate culture medium, culturing at 30 deg.C for 4 d. Respectively transferring the activated trichoderma harzianum and metarhizium anisopliae seeds to respective liquid fermentation culture media.

And (3) fermenting and culturing trichoderma harzianum liquid: the culture temperature is 25-30 ℃, the rotation speed is 150-. The culture medium consists of corn starch 15-20 weight portions, bean dregs 30-40 weight portions, glucose 15-20 weight portions, sodium chloride 2-3 weight portions, sodium dihydrogen phosphate 0.4-0.5 weight portion, ammonium sulfate 4-6 weight portions, calcium carbonate 2-4 weight portions, and magnesium sulfate 0.4-0.5 weight portion.

Carrying out liquid fermentation culture on metarhizium anisopliae: the culture temperature is 28-30 ℃, the rotation speed is 180-. The culture medium consists of 15-20 parts of glucose, 8-10 parts of yeast extract, 4-6 parts of ammonium sulfate, 0.4-0.6 part of potassium dihydrogen phosphate, 0.1-0.15 part of magnesium sulfate and 0.1 part of ferrous sulfate.

And 4, step 4: transferring the mother liquor obtained in the step 3 to a solid culture medium.

And (3) solid fermentation culture of trichoderma harzianum: the inoculation amount is 18-22%, the temperature is 25-30 ℃, the humidity is 70-80%, the culture is carried out for 2 weeks, the initial pH value of the culture is 6.5-7.0, and the culture is turned and thrown once every 48 hours. The solid culture medium is composed of 35-40 parts of wheat bran, 25-30 parts of rice hulls, 25-32 parts of straws, 4-6 parts of soybean meal, 1-3 parts of potassium dihydrogen sulfate and 2-4 parts of magnesium sulfate, and the raw materials of the culture medium are crushed by a superfine crusher, and the thickness of the culture medium is 4 cm.

And (3) carrying out solid fermentation culture on metarhizium anisopliae: the inoculation amount is 20-22%, the temperature is 25-30 ℃, the humidity is 75-80%, the culture is carried out for 2 weeks, the initial pH value of the culture is 6.5-7.0, and the culture is turned and thrown once every 48 hours. The solid culture medium is composed of 50-70 parts of wheat bran, 18-22 parts of corn flour and 8-12 parts of soybean meal, and the raw materials of the culture medium are crushed by a superfine crusher, and the thickness of the culture medium is 4 cm.

And 5: and (5) drying the solid culture medium containing the trichoderma harzianum and the solid culture medium containing the metarhizium anisopliae in the step (4) at low temperature, and mixing according to the viable count ratio of 1:5 to obtain the composite fungal powder.

Step 6: and (3) mixing the bacterial powder obtained in the step (2) and the fungal powder obtained in the step (5) according to the viable count ratio of 50:1 to obtain the composite bacterial powder for the Chinese yam.

Example 2

The embodiment provides composite bacterial powder for Chinese yam, which is different from the embodiment 1 in that in the step 2, bacillus subtilis, bacillus laterosporus, bacillus polymyxa and bacillus thuringiensis fermentation liquor are mixed according to the ratio of viable count of 20:1:1: 3. The rest steps are the same as those in embodiment 1, and are not described herein again.

Example 3

The embodiment provides a composite bacterial powder for Chinese yam, which is different from embodiment 1 in that in step 2, bacillus subtilis, bacillus laterosporus, bacillus polymyxa and bacillus thuringiensis fermentation liquor are mixed according to a viable count ratio of 10:1:1:1, and the rest steps are the same as embodiment 1 and are not repeated.

Example 4

The embodiment provides a composite bacterial powder for Chinese yam, which is different from embodiment 1 in that bacillus subtilis, bacillus laterosporus, bacillus polymyxa and bacillus thuringiensis fermentation liquor in step 2 are mixed according to the viable count ratio of 200:1:1:10, and other steps are the same as embodiment 1 and are not repeated.

Example 5

The embodiment provides a composite bacterial powder for Chinese yam, which is different from embodiment 1 in that bacillus subtilis, bacillus laterosporus, bacillus polymyxa and bacillus thuringiensis fermentation liquor in step 2 are mixed according to the viable count ratio of 30:2:2:7, and other steps are the same as embodiment 1 and are not repeated.

Example 6

The embodiment provides a composite bacterial powder for Chinese yam, which is different from embodiment 1 in that in step 2, bacillus subtilis, bacillus laterosporus, bacillus polymyxa and bacillus thuringiensis fermentation liquor are mixed according to a viable count ratio of 25:2:2:5, and the rest steps are the same as embodiment 1 and are not repeated.

Example 7

The embodiment provides a composite bacterial powder for Chinese yam, which is different from embodiment 1 in that bacillus subtilis, bacillus laterosporus, bacillus polymyxa and bacillus thuringiensis fermentation liquor in step 2 are mixed according to the viable count ratio of 40:1:1:10, and other steps are the same as embodiment 1 and are not repeated.

Example 8

The present embodiment provides a composite bacterial powder for Chinese yam, which is different from embodiment 1 in that the solid medium containing trichoderma harzianum and the solid medium containing metarhizium anisopliae in step 5 are dried at low temperature and then mixed according to the viable bacteria ratio of 2:1, and the rest steps are the same as embodiment 1 and are not described herein again.

Example 9

The present embodiment provides a composite bacterial powder for Chinese yam, which is different from embodiment 1 in that in step 5, a solid medium containing trichoderma harzianum and a solid medium containing metarhizium anisopliae are subjected to low temperature drying and then mixed according to a viable bacteria ratio of 1:2, and the remaining steps are the same as embodiment 1 and are not repeated herein.

Example 10

The present embodiment provides a composite bacterial powder for Chinese yam, which is different from embodiment 1 in that the solid medium containing trichoderma harzianum and the solid medium containing metarhizium anisopliae in step 5 are dried at low temperature and then mixed according to the viable bacteria ratio of 1:20, and the rest steps are the same as embodiment 1 and are not described herein again.

Example 11

The present embodiment provides a composite bacterial powder for yam, which is different from embodiment 1 in that in step 6, bacterial powder and fungal powder are mixed according to a viable bacteria ratio of 100:1, and the remaining steps are the same as embodiment 1, and are not described herein again.

Example 12

The present embodiment provides a composite bacterial powder for yam, which is different from embodiment 1 in that in step 6, bacterial powder and fungal powder are mixed according to a viable bacteria ratio of 2:1, and the remaining steps are the same as embodiment 1, and are not described herein again.

Comparative example 1

The comparative example provides composite bacterial powder for Chinese yam, which is different from example 1 in that bacterial powder and fungal powder are mixed according to a ratio of 1:2 in step 6, and the rest steps are the same as example 1 and are not repeated.

Comparative example 2

The comparative example provides composite bacterial powder for Chinese yam, which is different from example 1 in that the bacterial powder only comprises bacillus subtilis, bacillus laterosporus and bacillus polymyxa, the viable count ratio is 50:1:1, and the rest raw materials and preparation steps are the same as those in example 1 and are not repeated.

Comparative example 3

The comparative example provides composite bacterial powder for Chinese yam, which is different from the example 1 in that the fungal bacterial powder only contains trichoderma harzianum, and the rest raw materials and preparation steps are the same as those in the example 1 and are not repeated.

Comparative example 4

This comparative example provides a compound fungus powder for chinese yam, and it is different from example 1, only contains bacterial powder in the compound fungus powder, does not contain fungi powder, and other raw materials and preparation steps are the same with example 1, and no longer describe here.

Comparative example 5

This comparative example provides a compound fungus powder for chinese yam, and it is different from example 1, only contains fungus powder in the compound fungus powder, does not contain bacterial fungus powder, and other raw materials and preparation steps all are the same with example 1, and no longer describe here.

Test example 1

The Chinese yams obtained in the examples 1 to 12 and the comparative examples 1 to 5 are activated by the composite bacterial powder and then inoculated at 4 corner points around a PDA culture medium plate, colletotrichum gloeosporioides, dioscorea opposita, fusarium solani and fusarium oxysporum are respectively inoculated at the center of the PDA culture medium plate inoculated with the composite bacteria, only pathogenic bacteria are inoculated on a blank group of PDA culture medium, the blank group of PDA culture medium is placed in an incubator at 35 ℃ for culture, the steps are repeated for 3 times, and after 3 days, the bacteriostasis rate is measured. The bacteriostatic rate was calculated by the following formula (1).

The bacteriostatic ratio (%) is (blank group colony diameter-treated group colony diameter)/blank group colony diameter × 100%

Formula (1)

TABLE 1 bacteriostatic effect of composite bacterial powder for Chinese yam on various pathogenic bacteria

As can be seen from table 1, the composite bacterial powders for yam provided in examples 1 to 12 all have the effect of inhibiting the growth of pathogenic bacteria such as colletotrichum gloeosporioides, diaphora dioscoreae, fusarium solani, fusarium oxysporum and the like, and have the best effect of inhibiting fusarium solani, which indicates that the composite bacterial powder for yam provided by the present invention has a good disease prevention effect. The compound bacterial powder for Chinese yam provided by the embodiment 1 has the best bacteriostasis rate, so that the compound bacterial fertilizer for Chinese yam provided by the following embodiments are all prepared according to the bacterial powder proportion in the embodiment 1.

Example 13

The embodiment provides a compound bacterial fertilizer for Chinese yam, which is prepared by uniformly mixing 400g of the compound bacterial powder for Chinese yam obtained in the embodiment 1, 200g of potassium fulvate, 200g of algal polysaccharide and 20g of chitosan oligosaccharide.

Example 14

The present embodiment provides a composite bacterial fertilizer for yam, and the difference in example 13 is that 200g of composite bacterial powder for yam, 500g of potassium fulvate, 100g of algal polysaccharide and 20g of chitosan oligosaccharide, and the remaining steps are the same as those in example 12, and are not described herein again.

Example 15

The embodiment provides a compound bacterial fertilizer for Chinese yam, and the difference of the embodiment 13 is that 500g of compound bacterial powder for Chinese yam, 200g of potassium fulvate, 100g of algal polysaccharide and 20g of chitosan oligosaccharide, and the rest steps are the same as those in the embodiment 12, and are not repeated herein.

Comparative example 6

The comparative example provides a compound bacterial fertilizer for Chinese yam, and 820g of the compound bacterial powder for Chinese yam obtained in example 1 is taken.

Comparative example 7

The comparative example provides a compound bacterial fertilizer for Chinese yam, 500g of the compound bacterial powder for Chinese yam obtained in the example 1 is uniformly mixed with 320g of potassium fulvate, and the compound bacterial fertilizer for Chinese yam is obtained.

Comparative example 8

The comparative example provides a compound bacterial fertilizer for Chinese yam, 500g of the compound bacterial powder for Chinese yam obtained in the example 1, 200g of potassium fulvic acid and 120g of algal polysaccharide are uniformly mixed to obtain the compound bacterial fertilizer for Chinese yam.

Comparative example 9

The comparative example provides a compound bacterial fertilizer for Chinese yam, and the compound bacterial powder for Chinese yam obtained in example 1 is 220g, potassium fulvate 200g, algal polysaccharide 200g and chitosan oligosaccharide 200g, and the compound bacterial fertilizer for Chinese yam is obtained by uniformly mixing.

Test example 2

The Chinese yams obtained in the examples 13 to 15 and the comparative examples 6 to 9 are uniformly mixed with 0.43g of the compound bacterial fertilizer and sieved fine soil, then the mixture is loaded into a flowerpot, 3 Chinese yam seedlings are planted in each pot, the mixture is placed into a sunlight greenhouse, after 5 days, the 2 nd larvae of the wireworms, the 3 rd larvae of the cutworms and the 2 nd larvae of the scarab (namely grubs) are respectively inoculated for 4 heads to the surface of soil, the test insects which can not enter the soil for 24 hours are replaced, the steps are repeated for 5 times, and the survival condition of each larva is checked after 5 days of inoculation. The planting density of the Chinese yam is 7000 plants/mu, and the application amount of the composite bacterial manure for the Chinese yam is 1 kg/mu, namely 0.143 g/plant; the blank did not apply any compound bacterial manure as a control.

TABLE 2 inhibitory Effect of Compound bacterial manure for Yam against soil insects

Test example 3

The composite bacterial manure for yam obtained in examples 13 to 15 and comparative examples 6 to 9 was used when yam was planted in spring in Yao Wen county, and was applied in the following manner:

bottom application: 1.5 kg/mu of the compound bacterial fertilizer is mixed with fine soil uniformly and then applied into the planting furrows.

Dressing: after seedling emergence, the compound bacterial fertilizer is applied for 5 times, wherein the compound bacterial fertilizer is applied for 5 times in a monthly manner at 500 g/mu/time.

The blank example is the case without applying the compound bacterial manure for the Chinese yam.

The yam obtained in test example 3 was subjected to the following measurements:

in the picking season, fresh weight, length, stem thickness and yield of yam were measured, and the results are shown in table 3.

TABLE 3 influence of composite bacterial manure for yam on yam yield

Item	Fresh weight (kg/plant)	Length (cm)	Diameter (cm)	Yield (kg/mu)
					Blank example	0.27	62.8	1.83	1520
Example 13	0.41	72.6	2.23	2172
					Example 14	0.38	71.9	2.18	1892
Example 15	0.40	72.2	2.20	1980
					Comparative example 6	0.34	69.0	2.09	1703
Comparative example 7	0.33	68.3	2.03	1629
					Comparative example 8	0.36	69.2	2.11	1732
Comparative example 9	0.33	67.9	2.01	1683

The influence on the disease index of the overground part of the Chinese yam is preliminarily graded in the following grading mode:

level 0: the whole plant leaves have no disease spots;

level 1: slightly, the lower leaves have scabs, and the upper leaves have no scabs;

and 2, stage: the lower part of the plant has more scabs, the upper part of the plant has less scabs, and the leaf falling rate of the plant is 1 to 30 percent;

and 3, level: serious, the leaf disease has more spots, and the leaf falling rate of the plants is 30 to 90 percent;

4, level: and (4) death, wherein the leaf falling rate of the plants reaches over 90 percent or the plants die.

The results of statistical calculations on the rated data are shown in table 4.

And (4) carrying out statistical calculation after grading, wherein the calculation mode is shown as a formula (2) and a formula (3).

TABLE 4 influence of composite bacterial manure for Chinese yam on disease index of overground part

Treatment of	Index of disease condition	Control effect (%)
			Blank example	48.2
Example 13	14.3	70.33
			Example 14	15.6	67.63
Example 15	14.9	69.09
			Comparative example 6	17.9	62.86
Comparative example 7	19.3	59.96
			Comparative example 8	17.1	64.52
Comparative example 9	19.8	58.92

The lesion length of yam was measured and the percentage of lesions was calculated, the results are shown in table 5.

TABLE 5 influence of composite bacterial manure for Chinese yam on root rot of Chinese yam

Item	Length of lesion	Percentage of lesion (%)
			Blank example	21.4	34.08
Example 13	4.2	5.79
			Example 14	4.8	6.68
Example 15	4.4	6.09
			Comparative example 6	6.4	9.28
Comparative example 7	6.7	9.81
			Comparative example 8	5.9	8.53
Comparative example 9	7.1	10.46

The number of rhizosphere microorganisms of yam soil was measured, and the measurement results are shown in table 6.

TABLE 6 influence of composite bacterial manure for Chinese yam on microbial count of root of Chinese yam soil

The yam appearance quality is counted, the appearance quality is mainly observed from deformity, defects, uniform length and thickness, color and surface smoothness, and the statistical result is shown in table 7:

TABLE 7 influence of composite bacterial manure for Chinese yam on appearance and quality of Chinese yam

Item	Distorted shape	Number of defects	Length and thickness uniformity	Degree of color and luster	Surface smoothness
						Blank example	Much more	Much more	Irregular and irregular	Dark colour	Is coarser than
Example 13	Very few	Is almost none	Uniformity	Bright colour	Smooth and smooth
						Example 14	Chinese character shao (a Chinese character of 'shao')	Chinese character shao (a Chinese character of 'shao')	Is more uniform	The color is brighter	Is smoother
Example 15	Very few	Is almost none	Uniformity	Bright colour	Is smooth and smooth
						Comparative example 6	Less	Is less	Is more uniform	Slightly brighter color	Is smoother
Comparative example 7	Is less	Is less	Is relatively uniform	Slightly darker colour	Is relatively smooth
						Comparative example 8	A little bit	Chinese character shao (a Chinese character of 'shao')	Is relatively uniform	The color is brighter	Is smoother
Comparative example 9	Is less	Less	Is relatively uniform	Slightly darker colour	Is relatively smooth

As can be seen from Table 2, the number of live insects of cutworms, wireworms and grubs is reduced after the compound bacterial fertilizer for Chinese yam is applied for 5 days, which shows that the compound bacterial fertilizer for Chinese yam provided by the invention can kill pests. As can be seen from Table 3, the composite bacterial fertilizer for Chinese yam provided by the invention can promote the growth and expansion of Chinese yam tuberous roots and dry matter accumulation, and can improve the yield of Chinese yam. Real-time observation shows that the surface brightness of the Chinese yam tuber root applying the composite bacterial manure for Chinese yam is better and the root hair is more. As can be seen from the data in tables 4, 5 and 6, the composite bacterial fertilizer for yam provided in the examples can reduce the yam infection degree in different degrees, increase the bacterial content in soil, inhibit the growth of soil fungi and reduce the number of soil-borne pathogenic bacteria. As can be seen from table 7, after the composite bacterial manure for yam provided in the examples was applied, the yam tuber had good color, less distortion, uniform length and thickness, and good appearance quality.

The compound bacterial fertilizer for the Chinese yam provided by the invention can prevent plant diseases and insect pests, promote plant growth, improve the quality and the yield of the Chinese yam, and is suitable for large-scale popularization and application.

Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. The compound bacterial fertilizer for the Chinese yam is characterized by comprising 400g of compound bacterial powder for the Chinese yam, 200g of potassium fulvate, 200g of algal polysaccharide and 20g of chitosan oligosaccharide;

the composite bacteria powder for the yam consists of bacteria powder and fungus powder according to the viable bacteria ratio of 50: 1;

the bacterial powder consists of bacillus subtilis, bacillus laterosporus, bacillus polymyxa and bacillus thuringiensis according to the ratio of viable count of 50:1:1: 10;

the fungus powder is composed of trichoderma harzianum and metarhizium anisopliae according to the viable count ratio of 1: 5.