CN112674116A - Compound microbial agent for preventing and treating scab of crops and preparation and use methods thereof - Google Patents
Compound microbial agent for preventing and treating scab of crops and preparation and use methods thereof Download PDFInfo
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Abstract
The microbial inoculum has strong pertinence to crop bacterial scab, reasonably utilizes the mutual synergistic action among microorganisms, improves the rhizosphere ecological environment of crops by utilizing the life activity of active microorganisms contained in the microbial inoculum, and converts nutrient elements which can not be directly utilized by the crops in soil into quick-acting nutrients which can be utilized by the crops; in the aspect of disease treatment, the microbial agent adopts the principle of bacteria treatment, utilizes antibiotic substances secreted by antagonistic strains to inhibit the growth of pathogenic bacteria in soil, ensures that the pathogenic bacteria can not reach pathogenic quantity in the soil, so that the crops can not be infected, the diseases of the crops can be prevented, and the microbial agent has strong adaptability and good stability, can effectively antagonize potato scab, tomato-pepper scab and winter jujube scab, effectively reduces the pesticide residue, can not cause pollution to the environment, and has the functions of fixing nitrogen, dissolving phosphorus and potassium, stimulating the growth of the plants, improving the quality of the crops and the like, thereby achieving the purposes of yield increase and high yield.
Description
Technical Field
The invention belongs to the technical field of microbial fertilizers, and particularly relates to a compound microbial agent for preventing and treating crop scab and a preparation method and a use method thereof.
Background
Scab, english: scabs (tied over wounds or sores); spot disease (of the plant). Scab is a common disease in crop planting and is divided into bacterial type and fungal type, wherein the bacterial type is mainly caused by Streptomyces and Xanthomonas (Xanthomonas) in actinomycetes; the fungi are mainly caused by Elsinoe and Elsinoe. At present, the scab of crops is mostly treated by chemical pesticide control in China, pesticide abuse causes very serious environmental problems, pathogenic bacteria generate drug resistance, in addition, the hardening and pollution of soil are aggravated by excessive use of chemical fertilizers, the quality of the soil is reduced and degraded, the organic matter content of the soil is seriously lower than the average level of the whole world, the harmfulness of heavy metals in the soil is increased, and the sustainable development of agriculture in China is harmed. The compound microbial agent adopts the principle of 'treating bacteria by bacteria' to aim at the main harmful bacteria causing scab, namely streptomyces scab, xanthomonas campestris and pseudomonas cepacia, has the advantages of targeting, safety and no residue, optimizing the microecological balance of crops and soil rhizosphere, improving the absorption and utilization rate of the crops on nutrients, and enhancing the stress resistance of the crops, such as drought resistance, low temperature resistance, high salt resistance and the like. In particular, the microbial liquid inoculant has the advantages of convenient use, obvious growth promotion and stress resistance effects and the like.
The potato scab is a common crop disease, and is an important soil-borne disease caused by streptomyces spp in plant pathogenic actinomycetes, wherein streptomyces scab, streptomyces galileoparus and streptomyces oedematus are taken as main bacteria, bacteria-bearing fertilizers and diseased potatoes are main initial infection sources, germs mainly infect tubers, invade and infect the tubers from skin pores and wounds, mainly damage the epidermis of potato tubers, cause tissue necrosis around the infection point of the epidermis, form flat spots, bulges or sunken spots on the potato tubers, and influence the commercial value of the potato scab. At present, chemical prevention and agricultural prevention methods are adopted in severe areas, no very effective medicament can be used in the aspect of chemical prevention, and environmental pollution and drug resistance are easily caused by improper use; agricultural control cannot control the disease fundamentally; the breeding and application of disease-resistant varieties are economic and effective measures, but no disease-resistant variety can be used at present; the biological control method does not pollute the environment, has no toxicity to people and livestock, has strong pertinence, is a fundamental measure for controlling soil-borne diseases, and is a fundamental way for solving the problem of soil pollution caused by excessive use of pesticide and chemical fertilizers. At present, China reports various biocontrol strains with control effect on potato scab, wherein bacillus (Bacillus spp.) is the majority. Due to the diversity and synchronous evolution characteristics of pathogenic bacteria, the adopted single antagonistic strain cannot achieve ideal control effect, so that a plurality of biological functions of the biocontrol bacteria can play a better role only by depending on the synergistic effect of two or more strains of bacteria.
Tomato-pepper scab is a common disease of tomatoes and is also one of important diseases of peppers. Tomatoes mainly harm the stem, leaves and fruits. The diseased leaves appear small water-soaked spots on the backs of the leaves in the early stage, gradually expand to be nearly round or are connected into irregularly shaped tawny diseased spots, and are dry and crisp in the later stage. The stem part firstly appears water-soaked chlorosis spots, and then expands vertically to be oblong and black brown disease spots with slightly concave centers; the surfaces of diseased fruits have water-soaked chlorosis spots which gradually expand, oil is soaked in the surface of the diseased fruits and brightens, then the diseased fruits are yellow brown or black brown, cork and nearly round rough dead spots, some of the diseased fruits are mutually connected to form irregular large spots, and the fruits are easy to drop when the joints of the fruit stalks and the fruits are damaged.
The scab of hot pepper mainly damages leaves, stems and fruits. A plurality of round or irregular dark green to yellow brown spots appear in the early stage after the leaves are infected with diseases, sometimes the spots appear in the ring, the back of the leaves are slightly raised and are in a blister shape, the front of the leaves is slightly concave, the spots after the stems are infected with the diseases are irregular strip spots or patches, round or long round dark green spots appear after the fruits are infected with the diseases, the diameter is about 0.5 cm, the edges are slightly raised, the surface is rough, and rotten fruits are caused.
The scab of winter jujube is also called ulcer disease, which can damage the jujube head, jujube hanger and leaves of winter jujube tree, to cause the blackening and drying of jujube head, the separation of jujube hanger and the falling of leaves, finally to cause the flower bud not to be formed or fallen, to seriously affect the fruit setting rate of winter jujube, and the disease is very serious in the main domestic winter jujube production area and becomes a natural enemy of winter jujube production.
In view of the above-mentioned problems, there is a need for improvement.
Disclosure of Invention
The technical problems solved by the invention are as follows: the microbial inoculum is characterized by strong pertinence to crop bacterial scab, reasonably utilizing the mutual synergistic action among microorganisms, improving the rhizosphere ecological environment of crops by utilizing the life activity of active microorganisms contained in the microbial inoculum, and converting nutrient elements which can not be directly utilized by the crops in soil into quick-acting nutrients which can be utilized by the crops; in the aspect of disease treatment, the microbial agent adopts the principle of bacteria treatment, utilizes antibiotic substances secreted by antagonistic strains to inhibit the growth of pathogenic bacteria in soil, ensures that the pathogenic bacteria can not reach pathogenic quantity in the soil, so that crops can not be infected, the disease of the crops can be prevented, the adaptability is strong, the stability is good, selected microorganisms can coexist in a symbiotic manner, the potato scab, tomato-pepper scab and winter jujube scab can be effectively antagonized, the pesticide residue can be effectively reduced, the environment can not be polluted, and the microbial agent has the functions of fixing nitrogen, dissolving phosphorus and potassium, stimulating the growth of the crops, improving the quality of the crops and the like, so that the purposes of increasing the yield and increasing the yield are achieved.
The technical scheme adopted by the invention is as follows: the compound microbial agent for preventing and treating the scab of the crops is liquid, and comprises the following active components in parts by weight:
55-75 parts of beneficial microbial bacteria liquid, wherein the beneficial microbial bacteria liquid is formed by combining 10-20 parts of small golden streptomyces bacteria liquid, 5-10 parts of bacillus polymyxa bacteria liquid, 15-20 parts of bacillus subtilis liquid, 10-15 parts of bacillus amyloliquefaciens liquid and 5-10 parts of bacillus aryabhattai bacteria liquid; the content of viable bacteria in the beneficial microorganism liquid is more than or equal to 2.0 multiplied by 109CFU/ml；
5-10 parts of polyglutamic acid;
15-35 parts of biological organic matters;
0.3-0.5 part of melatonin.
The molecular weight of the polyglutamic acid is 5-200 kilodaltons, and the polyglutamic acid is polyglutamic acid sodium salt.
The biological organic matter is one or the combination of two of biological humic acid liquid, molasses and yeast extract.
The preparation method of the compound microbial agent for preventing and treating the scab of the crops comprises the following steps: the composite microbial agent is prepared by mixing five bacterial liquids, namely small golden streptomyces bacteria liquid, bacillus polymyxa bacteria liquid, bacillus subtilis liquid, bacillus amyloliquefaciens liquid and bacillus aryabhattai liquid, which are prepared in proportion according to parts, adding melatonin, polyglutamic acid and biological organic matters which are prepared according to parts, and uniformly mixing.
The compound microbial agent is mainly used for bacterial scab caused by streptomyces scab, streptomyces galileoparus, streptomyces oedemansicus, xanthomonas campestris and pseudomonas cepacia on crops, and is used for any one or more of improving the yield of crops, improving the quality of the crops and enhancing the stress resistance of the crops;
the use method of the compound microbial agent comprises the following steps:
firstly, spraying on leaf surfaces, namely spraying 2 kilograms per mu 7 to 15 days before flowering;
secondly, seed dressing, wherein the seed dressing amount of the compound microbial agent is 1/100 of the weight of the seeds, and the compound microbial agent is uniformly sprayed on the surfaces of the seeds;
thirdly, dipping the roots, adhering the compound microbial inoculum to the roots of the crops when the crops are transplanted, and planting the crops into soil.
Wherein the compound microbial agent is used for preventing and treating potato scab caused by streptomyces scabies, streptomyces galileoparus and streptomyces coelomis; the method for preventing and treating potato scab comprises the following steps: applied to crops by spraying or dressing; the seed dressing amount is 1/100 of the weight of the potato seeds, and the diluted seed dressing agent is uniformly sprayed on the surfaces of the seeds; for early-maturing potato varieties, the spraying is carried out 7-10 days before the first time of flower blooming, and the spraying is carried out 7-10 days after the second time of flower blooming.
The compound microbial agent is used for preventing and treating tomato-pepper scab caused by Xanthomonas campestris scab variety; the method for preventing and treating tomato-pepper scab comprises the following steps: soaking seeds in warm water at about 52 ℃ for 10 minutes before culturing seedlings of tomatoes and hot peppers, soaking the seeds in the compound microbial agent for 15 minutes, and accelerating germination and sowing; when transplanting, the roots are dipped with the compound microbial agent, 2 kilograms per mu are sprayed once 10 days before flower buds emerge, and the spraying is carried out once again at 2 kilograms per mu in the fruit setting period.
The compound microbial agent is used for preventing and treating winter jujube scab caused by xanthomonas campestris and pseudomonas cepacia; the method for preventing and treating winter jujube scab comprises the following steps: spraying 2 kilograms/mu 15 days before the bud emerges, spraying once every five days, and spraying twice continuously; spraying the fertilizer once at the fruit setting period of 2 kilograms per mu.
Compared with the prior art, the invention has the advantages that:
1. the compound microbial agent is formed by compounding melatonin, five beneficial microbial liquid, biological organic matters and polyglutamic acid which are prepared according to the proportion, is different from common chemical fertilizers and chemical pesticides, improves the rhizosphere ecological environment of crops by utilizing the life activity of active microorganisms contained in the compound microbial agent, and converts nutrient elements which can not be directly utilized by the crops in soil into quick-acting nutrients which can be utilized by the crops; in the aspect of disease treatment, selected microorganisms coexist in symbiosis, and the microbial inoculant adopts the principle of treating bacteria by bacteria, utilizes antibiotic substances secreted by antagonistic strains to inhibit the growth of pathogenic bacteria in soil, so that the pathogenic bacteria cannot reach pathogenic quantity in the soil, and thus crops cannot be infected and the crops are prevented from being attacked; the microbial inoculum has strong adaptability and good stability, and microorganisms contained in the microbial inoculum can quickly grow on stems, leaves and roots of crops after being sprayed, secrete growth-promoting substances to stimulate the growth of the crops, improve the quality of the crops and achieve the purposes of increasing yield and high yield;
2. the chemical name of melatonin used in the microbial inoculum is N-acetyl-5-methoxytryptamine, the melatonin is an indole micromolecule compound, the compound is safe to human and animals, and compared with the traditional pesticide, the melatonin is an ideal plant protective agent. Research shows that the plant melatonin not only participates in regulation of plant growth and development, flowering, maturation and aging, but also participates in various stress responses such as salt stress, drought stress, high-temperature stress and low-temperature stress. The external application of melatonin can improve the heat resistance and disease tolerance of plants, and leaf spots of crops such as potatoes, tomatoes, peppers and the like treated by the melatonin are obviously reduced compared with those of the crops without the melatonin. The invention is used for preventing and treating crop scab according to the characteristics of melatonin, improves the drought resistance and salt tolerance of crops through the melatonin and beneficial microbial flora, and leads bacterial scab pathogenic bacteria of crops to be effectively antagonized;
3. streptomyces aureofaciens used in the microbial inoculum is taken as one of microbial pesticides, mainly produces kasugamycin and a plurality of enzymes, and in a new bioactive substance produced by microbes, the yield of the streptomyces accounts for 74%; the main metabolite kasugamycin of the streptomyces parvus has the action mechanism of an esterase system interfering the metabolism of pathogenic bacteria amino acid, destroying the biosynthesis of protein, inhibiting the growth of hypha and causing cell granulation, so that the pathogenic bacteria lose the reproductive and infection capacity, thereby achieving the purposes of killing the pathogenic bacteria and preventing and treating diseases. Meanwhile, the streptomyces parvus can quickly grow around stems, leaves and roots of crops by spraying, generates a large amount of antibiotics and macrolactin, and can effectively inhibit the growth of scab pathogenic bacteria;
4. the bacillus amyloliquefaciens used in the microbial inoculum can generate a plurality of physiological active substances and amino acid substances such as gibberellin, indoleacetic acid, cytokinin and the like, and the generated antibacterial substances mainly comprise a plurality of antibacterial active substances such as sweet protein, sweet-like protein, antibiotic peptide substances, iturin, surfactant, chitinase, antibiotics and the like, so that the growth of plant roots and plants is promoted, the disease resistance of the plants is enhanced, and the disease occurrence is indirectly reduced;
5. the bacillus aryabhattai and the bacillus subtilis which are used in the microbial inoculum are both indigenous bacteria in soil and plants, can generate spores with heat resistance and stress resistance, have remarkable biocontrol potential, can be used as root growth promoting microorganisms, can parasitize roots, leaves and plant surfaces, can quickly colonize plant root hairs, stems and leaves and other surfaces, and can prevent the establishment of pathogenic fungi and bacteria; in addition, the Bacillus aryabhattai can inhibit the growth of rhizopus, and has good effect of preventing root mold and root rot;
6. the polyglutamic acid used in the microbial inoculum is a special anionic high molecular polymer, the side chain contains a large number of carboxyl groups, the groups endow the microbial inoculum with special biological activity, and the polyglutamic acid can provide the membrane-clamping protection for the growth-promoting bacteria as an adhesive substance, so that beneficial bacteria form an external coating biological membrane system, thereby isolating the material exchange inside and outside the growth-promoting bacteria cells and achieving a balanced stable state.
Drawings
FIG. 1 is a comparison of leaves of tomato control group and experimental group;
FIG. 2 is a graph showing the growth of leaves and fruits of a control strain of Capsicum annuum according to the present invention;
FIG. 3 is a diagram showing the growth of leaves and fruits of a pepper experimental plant according to an embodiment of the present invention;
FIG. 4 is a comparison of flower buds of a control group and an experimental group of winter jujubes in an embodiment of the present invention;
FIG. 5 is a comparison graph of flowering and fruit setting of the winter jujube control group and the winter jujube experimental group in the embodiment of the invention.
Detailed Description
Embodiments of the present invention are described below.
The compound microbial agent for preventing and treating the scab of the crops comprises the following components:
the compound microbial agent is liquid, and comprises the following active components in parts by weight:
55-75 parts of beneficial microbial strain liquid: wherein the culture medium is formed by compounding and mixing five fermentation bacterial liquids, namely 10-20 parts of streptomyces aureofaciens bacterial liquid, 5-10 parts of bacillus polymyxa bacterial liquid, 15-20 parts of bacillus subtilis bacterial liquid, 10-15 parts of bacillus amyloliquefaciens bacterial liquid and 5-10 parts of bacillus aryabhattai bacterial liquid; the content of viable bacteria in the beneficial microorganism liquid is more than or equal to 2.0 multiplied by 109CFU/ml；
5-10 parts of polyglutamic acid, wherein the molecular weight of the polyglutamic acid is 5-200 ten thousand daltons, and the polyglutamic acid is a polyglutamic acid sodium salt;
15-35 parts of biological organic matters, wherein the biological organic matters are one or a combination of two of biological humic acid liquid, molasses and yeast extract;
0.3-0.5 part of melatonin.
Streptomyces aureofaciens used in the microbial inoculum is taken as one of microbial pesticides, mainly producing kasugamycin and a plurality of enzymes, and in a new bioactive substance produced by microbes, the yield of the streptomyces accounts for 74%. The main metabolite kasugamycin of the streptomyces parvus has the action mechanism of an esterase system interfering the metabolism of pathogenic bacteria amino acid, destroying the biosynthesis of protein, inhibiting the growth of hypha and causing cell granulation, so that the pathogenic bacteria lose the reproductive and infection capacity, thereby achieving the purposes of killing the pathogenic bacteria and preventing and treating diseases. Meanwhile, the streptomyces parvus can quickly grow around stems, leaves and roots of crops by spraying, generates a large amount of antibiotics and macrolactin, and can effectively inhibit the growth of scab pathogenic bacteria.
The bacillus amyloliquefaciens used in the microbial inoculum can generate a plurality of physiological active substances and amino acid substances such as gibberellin, indoleacetic acid, cytokinin and the like, and the generated antibacterial substances mainly comprise a plurality of antibacterial active substances such as sweet protein, sweet-like protein, antibiotic peptide substances, iturin, surfactant, chitinase, antibiotics and the like, so that the growth of plant roots and plants is promoted, the disease resistance of the plants is enhanced, and the disease occurrence is indirectly reduced.
The bacillus aryabhattai and the bacillus subtilis which are used in the microbial inoculum are both the original bacteria in soil and plants, can generate spores with heat resistance and stress resistance, have remarkable biocontrol potential, can be used as root growth promoting microorganisms, can parasitize roots, leaves and plant surfaces, can quickly colonize plant root hairs, stems and leaves and other surfaces, and can prevent the establishment of pathogenic fungi and bacteria. In addition, the Bacillus aryabhattai can inhibit the growth of rhizopus and has good effect of preventing root mold and root rot.
The polyglutamic acid used in the microbial inoculum is a special anionic high molecular polymer, the side chain contains a large number of carboxyl groups, the groups endow the microbial inoculum with special biological activity, and the polyglutamic acid can provide the membrane-clamping protection for the growth-promoting bacteria as an adhesive substance, so that beneficial bacteria form an external coating biological membrane system, thereby isolating the material exchange inside and outside the growth-promoting bacteria cells and achieving a balanced stable state.
The biological organic matter in the microbial inoculum is biological humic acid liquid agent, and is high-quality organic matter.
The chemical name of melatonin in the microbial inoculum is N-acetyl-5-methoxytryptamine, the microbial inoculum is an indole micromolecule compound, the microbial inoculum has safety to human and animals, and compared with the traditional pesticide, the melatonin is an ideal plant protective agent. Research shows that the plant melatonin not only participates in regulation of plant growth and development, flowering, maturation and aging, but also participates in various stress responses such as salt stress, drought stress, high-temperature stress and low-temperature stress. The external application of melatonin can improve the heat resistance and disease tolerance of plants, and leaf spots of crops such as potatoes, tomatoes, peppers and the like treated by the melatonin are obviously reduced compared with those of the crops without the melatonin. The invention is based on the characteristics of melatonin, and is used for preventing and treating crop scab, and the drought resistance and salt tolerance of crops are improved through the melatonin and beneficial microbial flora, so that the bacterial scab pathogenic bacteria of the crops can be effectively antagonized. Through the synergistic effect of the melatonin and the rhizosphere salt-tolerant microbial community of the crops, a new way for improving the stress resistance of the crops by using a non-transgenic way is developed, and the international competitiveness in the aspect of agricultural scientific research of China is improved.
In a word, the compound combination of five bacteria solutions, melatonin, biological organic matters and sodium polyglutamate in the compound microbial agent is different from common chemical fertilizers and chemical pesticides, the ecological environment of the rhizosphere of crops is improved by using the life activity of active microorganisms contained in the compound microbial agent, and nutrient elements which cannot be directly utilized by the crops in soil are converted into quick-acting nutrients which can be utilized by the crops; in the aspect of disease treatment, selected microorganisms coexist in symbiosis, and the microbial agent adopts the principle of bacteria treatment, utilizes antibiotic substances secreted by antagonistic strains to inhibit the growth of pathogenic bacteria in soil, so that the pathogenic bacteria cannot reach pathogenic quantity in the soil, and thus crops cannot be infected and the crops are prevented from being attacked. The microbial inoculum has strong adaptability and good stability, and microorganisms contained in the microbial inoculum can quickly grow on stems, leaves and roots of crops after being sprayed, secrete growth-promoting substances to stimulate the growth of the crops, improve the quality of the crops and achieve the purposes of increasing yield and high yield.
Secondly, a preparation method of the compound microbial agent for preventing and treating the scab of the crops comprises the following steps:
the method comprises the following steps: the composite microbial agent is prepared by mixing five kinds of bacteria liquid, namely small golden streptomyces bacteria liquid, bacillus polymyxa liquid, bacillus subtilis liquid, bacillus amyloliquefaciens liquid and bacillus aryabhattai liquid, which are prepared in proportion according to parts, and then adding melatonin, polyglutamic acid and biological organic matters which are prepared according to parts.
The preparation method of the Streptomyces aureofaciens bacterial liquid in the compound microbial agent comprises the following steps:
selecting a kasugamycin high-yield strain streptomyces parvus oblique surface test tube seed, respectively scratching a plate by using a Gauss synthesis first agar culture medium for resuscitation, culturing for 36 hours at 28 ℃, selecting a single bacterial colony under the plate to inoculate the Gauss synthesis first liquid culture medium, performing shake-flask culture at 28 ℃ and 150r/min for 48 hours to form a large amount of hyphae, and preparing the streptomyces parvus liquid seed for later use; inoculating 2L of the shake flask seeds into a 200L seed tank filled with Gao's synthetic first culture medium, stirring at 28 ℃ for 180r/min, and allowing ventilation to be 1: 1 culture for 40 hours, inoculating the culture solution into a 2000L fermentation tank, stirring the mixture at the temperature of between 25 and 30 ℃ for 300r/min, and introducing air flow into a reactor with the ratio of 1: culturing for 5 days to collect the fermentation liquid, namely the Streptomyces aureofaciens bacterial liquid.
(2) The preparation method of the bacillus polymyxa bacterial liquid in the composite microbial agent comprises the following steps:
taking out the bacillus polymyxa preservation tube, respectively scratching a plate by using a nutrient gravy solid culture medium for resuscitation, and culturing for 48 hours at 35 ℃; selecting a single colony under a plate, inoculating the single colony to a nutrient broth solid culture medium, culturing for 48 hours in an incubator at 35 ℃, eluting lawn with 2000ml of sterile water, inoculating the colony to a 200L fermentation tank filled with a bacillus polymyxa liquid seed culture medium, stirring for 120r/min, ventilating for 200L/min in the first 10 hours, ventilating for 320L/min after 10 hours, culturing for 16-24 hours at 35 ℃, and concentrating until the total bacteria content is not lower than 20 hundred million CFU/ml, thus obtaining the bacillus polymyxa bacterial liquid.
(3) The preparation method of the bacillus aryabhattai bacterial liquid in the compound microbial agent comprises the following steps:
taking out the Bacillus aryabhattai preservation tube, respectively scratching a plate by using a nutrient broth solid culture medium for resuscitation, and culturing for 48 hours at 36 ℃; selecting a single colony under a flat plate, inoculating the single colony to a nutrient broth solid culture medium, culturing for 48 hours in an incubator at 36 ℃, eluting lawn with 2000ml of sterile water, inoculating the single colony to a 200L fermentation tank filled with an alder liquid seed culture medium, stirring for 120r/min, ventilating for 200L/min in the first 10 hours, ventilating for 320L/min after 10 hours, culturing for 16-24 hours at 36 ℃, and concentrating until the total bacteria content is not lower than 20 hundred million CFU/ml, thus obtaining the bacillus alder bacterial liquid.
(4) The preparation method of the bacillus amyloliquefaciens bacterial liquid in the composite microbial agent comprises the following steps:
taking out the inclined storage tube of the bacillus amyloliquefaciens, respectively scratching a plate by using nutrient gravy agar culture medium for resuscitation, culturing for 48 hours at 36 ℃, selecting a single colony under the plate to inoculate to the nutrient gravy agar culture medium, culturing for 24 hours in an incubator at 36 ℃, eluting the lawn in the plate by using 2000ml of sterile water, inoculating to a 200L fermentation tank containing the bacillus amyloliquefaciens liquid seed culture medium, stirring for 120r/min, wherein the ventilation volume is 200L/min in the first 12 hours, the ventilation volume is 320L/min after 12 hours, culturing for 16-24 hours at 36 ℃, and taking the bacillus amyloliquefaciens liquid seed as the liquid seed when the total bacteria content is not less than 10 hundred million CFU/ml; inoculating bacillus amyloliquefaciens strains into a sterilized 2000L fermentation tank; fermentation conditions are as follows: and (3) the aeration ratio is 1:0.6, the culture temperature is 37 ℃, the stirring revolution is 200rpm, the dissolved oxygen is not less than 30%, the pH value is not less than 6.5, and the fermentation broth, namely the bacillus amyloliquefaciens bacterial liquid, is collected after stirring culture for 15 hours.
(5) The preparation method of the bacillus subtilis liquid in the compound microbial agent comprises the following steps:
taking out the bacillus subtilis storage tube, respectively scratching a plate by using a nutrient gravy solid culture medium for resuscitation, and culturing for 24 hours at 36 ℃; selecting a single colony under a flat plate, inoculating the single colony to a solid culture medium filled with nutrient gravy, culturing for 24 hours in an incubator at 36 ℃, eluting lawn with 2000ml of sterile water, inoculating the single colony to a 200L fermentation tank filled with a liquid seed culture medium, stirring for 120r/min, ventilating for 200L/min in the first 10 hours, ventilating for 320L/min after 10 hours, culturing for 24-30 hours at 36 ℃, and concentrating until the total bacteria content is not lower than 20 hundred million CFU/ml, thus obtaining the bacillus subtilis liquid.
Thirdly, the use method of the compound microbial agent for preventing and treating the scab of the crops comprises the following steps:
the compound microbial agent is mainly used for bacterial scab caused by streptomyces scabiosus, streptomyces galileoparus, streptomyces oedemansi, xanthomonas campestris and pseudomonas cepacia on crops, and is used for any one or more of improving the yield of crops, improving the quality of crops and enhancing the stress resistance of crops.
The use method of the compound microbial agent comprises the following steps:
firstly, spraying on leaf surfaces, namely spraying 2 kilograms per mu 7 to 15 days before flowering;
secondly, seed dressing, wherein the seed dressing amount of the compound microbial agent is 1/100 of the weight of the seeds, and the compound microbial agent is uniformly sprayed on the surfaces of the seeds;
thirdly, dipping the roots, adhering the compound microbial inoculum to the roots of the crops when the crops are transplanted, and planting the crops into soil.
In particular to different application methods for different crops.
1. For potatoes: the compound microbial agent is used for preventing and treating potato scab caused by streptomyces scabies, streptomyces galileoparus and streptomyces coelomis. The method for preventing and treating potato scab comprises the following steps: applied to crops by spraying or dressing; seed dressing is carried out on potato seeds by using the microbial agent for preventing and treating the crop scab, the seed dressing amount is 1/100 of the weight of the potato seeds, and the diluted seed dressing is uniformly sprayed on the surfaces of the seeds; for early-maturing potato varieties, the spraying is carried out 7-10 days before the first time of flower blooming, and the spraying is carried out 7-10 days after the second time of flower blooming.
2. For tomatoes and peppers: the compound microbial agent is used for preventing and treating tomato-pepper scab caused by Xanthomonas campestris scab variety. The method for preventing and treating tomato-pepper scab comprises the following steps: soaking seeds in warm water at about 52 ℃ for 10 minutes before culturing seedlings of tomatoes and hot peppers, soaking the seeds in the compound microbial agent for 15 minutes, and accelerating germination and sowing; when transplanting, the roots are dipped with the compound microbial inoculum, the compound microbial inoculum is sprayed once for 10 days before the flower buds emerge, and the compound microbial inoculum is sprayed once again for 2 kilograms per mu in the fruit setting period.
3. Application to winter jujubes: the compound microbial agent is used for preventing and treating winter jujube scab caused by xanthomonas campestris and pseudomonas cepacia. The method for preventing and treating winter jujube scab comprises the following steps: spraying 2 kilograms/mu 15 days before the bud emerges, spraying once every five days, and spraying twice continuously; spraying the fertilizer once at the fruit setting period of 2 kilograms per mu.
Fourth, field test effect of compound microbial inoculum for preventing and treating scab
(1) For potatoes
In the control group of 100 mu, the potato seeds are mixed with 70 percent thiophanate methyl and talcum powder, and the microbial inoculum is not applied in the bud period. In an experimental group of 100 mu, potato seeds are mixed with the microbial inoculum and are sprayed once 7-10 days before blooming and 7-10 days after blooming.
The test and control groups were sampled at harvest time, respectively, with the sampling point being a single row 10 meters long. Observing the growth and premature senility conditions of the overground part, measuring the number of plants at a sampling point with the row length of 10 meters, the total number and weight of the potato blocks, the number of the potato blocks with diseases, the number of the potato blocks with goods, the number of the potato blocks without diseases, the number and weight of the potato blocks with goods, and calculating the disease resistance rate and the yield per mu.
One set of experiments: (a) the length of 10 meters is 55 plants, 11 sampling points are arranged, and 10 samples are compared;
(b) the length of 10 meters is 53 plants, 10 sampling points are set, and 10 samples are compared;
(c) the length of 10 meters is 57 plants with 12 sampling points, and the number of the sampling points is 12;
two groups of experiments were carried out: (a) the length of 10 meters is 53 plants, 10 sampling points are set, and 9 samples are compared;
(b) the length of 10 meters is 55 plants, 11 sampling points are arranged, and 10 samples are compared;
(c) the length of 10 meters is 56 plants, 11 sampling points are arranged, and 11 samples are compared;
three groups of experiments were performed: (a) the length of the plant is 10 meters, and the total length of the plant is 46 plants, and 9 sampling points are set, and 9 controls are set;
(b) the length of 10 meters is 43 plants, and 8 sampling points are arranged, and 9 samples are compared;
(c) the length of the 10 meters is 43 plants, and 9 sampling points are arranged, and the number of the samples is 8;
data statistics
| Group (McKen I early maturing) | Total number (number) | Disease-free potato (sweet potato) | Control rate (%) | Total yield (kilogram/mu) | Commercial potato yield (kilogram/mu) | Commercial potato rate (%) |
| Control group | 171 | 102 | 59.6％ | 2873 | 2510 | 87.37 |
| Test set (200 kg/mu) | 192 | 183 | 95.3 | 3140 | 2900 | 92.36 |
As can be seen from the table above, the control effect of the test group treated by 200 kg in the test group reaches 95.3%, which is 35.7% higher than that of the control group. The compound microbial agent obviously improves the commodity potato rate of potatoes, wherein the commodity potato rate of a 200 kg treatment test group is improved by 5 percent, the commodity potato selling price in this year is 2200 yuan/ton, but the non-commodity potato selling price is 1500 yuan/ton, the income per mu is increased by 673.5 yuan, and the economic benefit is considerable.
(2) For tomato-pepper
The pepper and the tomato are soaked by the compound microbial agent. Soaking seeds in warm water at about 52 ℃ for 10 minutes before culturing seedlings of tomatoes and hot peppers, soaking the seeds in the complex microbial inoculum for 15 minutes, and accelerating germination and sowing. When transplanting, the root is dipped with the microbial inoculum, the compound microbial inoculum is sprayed for 2 kilograms/mu 15 days before the bud emerges, the compound microbial inoculum is sprayed for one time every 5 days, the compound microbial inoculum is continuously applied for two times, and the compound microbial inoculum is sprayed for one time again at 2 kilograms/mu in the fruit setting period. As can be seen from the comparative graphs of the leaves of the tomato control group and the tomato experimental group shown in FIG. 1, the leaves of the tomato control group are withered and yellow and curly, and the leaves of the tomato experimental group are healthy; as can be seen from the growth chart of the leaves and fruits of the pepper control plant shown in FIG. 2, the control pepper plant has obvious lesion spots on the leaves and fruits; the growth chart of the leaves and fruits of the pepper experimental plant shown in FIG. 3 shows that the leaves of the experimental pepper plant are healthy and the fruits have no disease spots.
(3) For winter jujube
A compound microbial agent for preventing and treating crop scab is used for preventing and treating experimental test sites of winter jujube scab: dali county Van Jia Zhen Ju Fa
The using method and the experimental result are as follows: spraying 2 kilograms per mu about 15 days before the bud of winter jujube comes first, and spraying once every five days and twice continuously. Spraying the fertilizer once at the fruit setting period of 2 kilograms per mu.
As can be seen from the comparison graph of flower buds of the control group and the experimental group of the winter jujubes shown in fig. 4 and the comparison graph of flowering and fruit setting of the control group and the experimental group of the winter jujubes shown in fig. 5, the control group has relatively serious jujube orchard scab, the flowering and fruit setting period is delayed, the flower buds fall off, and the influence is large; after the experimental group is sprayed with the compound microbial inoculant, the flowering and fruit setting are early, and the development is good.
The tests show that said compound microbial preparation can effectively resist potato scab, tomato-hot pepper scab and winter jujube scab, can effectively reduce pesticide residue, does not produce pollution to environment, and has the functions of stimulating plant growth and improving crop quality, etc. so as to attain the goal of raising yield and raising yield.
The above-mentioned embodiments are merely preferred embodiments of the present invention, which are not intended to limit the scope of the present invention, and therefore, all equivalent changes made by the contents of the claims of the present invention should be included in the claims of the present invention.
Claims (8)
1. The compound microbial agent for preventing and treating the scab of the crops is characterized in that: the compound microbial agent is liquid, and comprises the following active components in parts by weight:
55-75 parts of beneficial microbial bacteria liquid, wherein the beneficial microbial bacteria liquid is formed by combining 10-20 parts of small golden streptomyces bacteria liquid, 5-10 parts of bacillus polymyxa bacteria liquid, 15-20 parts of bacillus subtilis liquid, 10-15 parts of bacillus amyloliquefaciens bacteria liquid and 5-10 parts of bacillus aryabhattai bacteria liquid; the content of viable bacteria in the beneficial microorganism liquid is more than or equal to 2.0 multiplied by 109CFU/ml；
5-10 parts of polyglutamic acid;
15-35 parts of biological organic matters;
0.3-0.5 part of melatonin.
2. The compound microbial agent for preventing and treating scab of crops as claimed in claim 1, wherein: the molecular weight of the polyglutamic acid is 5-200 ten thousand daltons, and the polyglutamic acid is sodium polyglutamate.
3. The compound microbial agent for preventing and treating scab of crops as claimed in claim 1, wherein: the biological organic matter is one or the combination of two of biological humic acid liquid, molasses and yeast extract.
4. The method for preparing the compound microbial agent for preventing and treating the scab of the crop according to claim 1, 2 or 3, wherein the method comprises the following steps: the method comprises the following steps: the composite microbial agent is prepared by mixing five bacterial liquids, namely small golden streptomyces bacteria liquid, bacillus polymyxa bacteria liquid, bacillus subtilis liquid, bacillus amyloliquefaciens liquid and bacillus aryabhattai liquid, which are prepared in proportion according to parts, adding melatonin, polyglutamic acid and biological organic matters which are prepared according to parts, and uniformly mixing.
5. The use method of the compound microbial agent for preventing and treating the scab of the crop as claimed in claim 4, wherein: the compound microbial agent is mainly used for bacterial scab caused by streptomyces scabiosus, streptomyces galileoparus, streptomyces oedemansi, xanthomonas campestris and pseudomonas cepacia on crops, and is used for any one or more of improving the yield of crops, improving the quality of crops and enhancing the stress resistance of crops;
the use method of the compound microbial agent comprises the following steps:
firstly, spraying on leaf surfaces, namely spraying 2 kilograms per mu 7 to 15 days before flowering;
secondly, seed dressing, wherein the seed dressing amount of the compound microbial agent is 1/100 of the weight of the seeds, and the compound microbial agent is uniformly sprayed on the surfaces of the seeds;
thirdly, dipping the roots, adhering the compound microbial inoculum to the roots of the crops when the crops are transplanted, and planting the crops into soil.
6. The use method of the compound microbial agent for preventing and treating the scab of the crop as claimed in claim 5, wherein: the compound microbial agent is used for preventing and treating potato scab caused by streptomyces scab, streptomyces galileoparus and streptomyces coelomis; the method for preventing and treating potato scab comprises the following steps: applied to crops by spraying or dressing; the seed dressing amount is 1/100 of the weight of the potato seeds, and the diluted seed dressing agent is uniformly sprayed on the surfaces of the seeds; for early-maturing potato varieties, the spraying is carried out 7-10 days before the first time of flower blooming, and the spraying is carried out 7-10 days after the second time of flower blooming.
7. The use method of the compound microbial agent for preventing and treating the scab of the crop as claimed in claim 5, wherein: the compound microbial agent is used for preventing and treating tomato-pepper scab caused by Xanthomonas campestris scab variety; the method for preventing and treating tomato-pepper scab comprises the following steps: soaking seeds in warm water at about 52 ℃ for 10 minutes before culturing seedlings of tomatoes and hot peppers, soaking the seeds in the compound microbial agent for 15 minutes, and accelerating germination and sowing; when transplanting, the roots are dipped with the compound microbial agent, 2 kilograms per mu are sprayed once 10 days before flower buds emerge, and the spraying is carried out once again at 2 kilograms per mu in the fruit setting period.
8. The use method of the compound microbial agent for preventing and treating the scab of the crop as claimed in claim 5, wherein: the compound microbial agent is used for preventing and treating winter jujube scab caused by xanthomonas campestris and pseudomonas cepacia; the method for preventing and treating winter jujube scab comprises the following steps: spraying 2 kilograms/mu 15 days before the bud emerges, spraying once every five days, and spraying twice continuously; spraying the fertilizer once at the fruit setting period of 2 kilograms per mu.
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