CN114315458A - Microbial source biological stimulator and preparation method thereof - Google Patents
Microbial source biological stimulator and preparation method thereof Download PDFInfo
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- CN114315458A CN114315458A CN202111392105.2A CN202111392105A CN114315458A CN 114315458 A CN114315458 A CN 114315458A CN 202111392105 A CN202111392105 A CN 202111392105A CN 114315458 A CN114315458 A CN 114315458A
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- photosynthetic bacteria
- nitrogen
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- porphyridium
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- 238000002360 preparation method Methods 0.000 title claims description 23
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- 230000000243 photosynthetic effect Effects 0.000 claims abstract description 50
- 241000195493 Cryptophyta Species 0.000 claims abstract description 48
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 38
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 22
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 19
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 18
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- 239000001963 growth medium Substances 0.000 claims description 35
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- 239000003795 chemical substances by application Substances 0.000 claims description 15
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Landscapes
- Fertilizers (AREA)
Abstract
The invention relates to the technical field of microorganisms and agriculture, and particularly discloses a microbial source biological stimulator which comprises the following components in parts by weight: 1 to 50 percent of porphyridium, 1 to 50 percent of nitrogen-fixing blue algae, 1 to 50 percent of photosynthetic bacteria, 1 to 10 percent of carbon source and 1 to 10 percent of nitrogen source. The microbial source biological stimulator is compounded by microalgae and photosynthetic bacteria, has the effects of fertilizer and pesticide, can promote the metabolism and growth of crops, and improves the disease resistance of the crops; the microbial source biological stimulator can effectively improve the absorption and stress resistance of nutrient substances by spraying on the leaf surfaces, can also improve the physical and chemical properties and community microorganisms of soil by spraying on the soil, and improves the yield and quality of crops.
Description
Technical Field
The invention belongs to the technical field of microorganisms and agriculture, and particularly relates to a microbial source biostimulant and a preparation method thereof.
Background
The use of pesticides and chemical fertilizers is always an important means for improving the crop yield, China is one of the countries with the largest production and application amount of agricultural chemicals in the world, and the excessive use of chemical fertilizers and pesticides brings negative effects to the environment and grain crops in China, such as pollution of surface water, underground water and near sea water, soil hardening, crop quality reduction, enhancement of pest resistance and the like. The development of zero-increase action of the usage amount of chemical fertilizers and pesticides by the organization in the rural area of agriculture is a long-term and difficult task. The method has the advantages of improving the fertilization and pesticide application mode, improving the utilization rate of pesticide and fertilizer, reducing unreasonable investment, and ensuring effective and safe supply of main agricultural products such as grains, and is a problem which needs to be solved urgently in agricultural sustainable development.
The plant biostimulant is a substance containing certain components and microorganisms, and when the plant biostimulant is applied to plants or roots, the components and the microorganisms play a role in stimulating the natural growth process of the plants, can regulate and control the growth and development of crops in a two-way mode, activate the secondary metabolism of the crops, induce the crops to generate a stress resistance mechanism, enhance the resistance and the immunity to adverse growth environment, activate soil nutrients, increase the oxygen supply capacity of the available nutrients of the soil, and further improve the utilization rate of the available components of the pesticide and fertilizer for the crops. Some microorganisms such as microalgae and bacteria can synthesize and accumulate various high-value-added biological activities with specific structure and function, including functional polysaccharide, lipid, functional protein, amino acid, natural active pigment, effective antibacterial components, phytohormone analogs and the like, and the high-value-added products can stimulate the generation of nonspecific active factors in plants, regulate the balance of endogenous hormones, improve the stress resistance of crops and enhance the absorption and utilization of nutrient substances. On the other hand, the microalgae and bacteria can improve the physicochemical property of soil, regulate the microbial community of the soil and enhance the soil fertility. Microalgae and bacteria are various in types, widely distributed, high in propagation speed and capable of being cultured in large scale, and become ideal sources for preparing novel powerful plant biological stimulators. The dominant algae species and bacteria are screened and cultured, the formula of the microbial source biological stimulator is optimized, the microbial source biological stimulator applicable to crops and soil property improvement is researched, and the method has important significance for agricultural planting.
Disclosure of Invention
The invention aims to provide a microbial source biological stimulator and a preparation method thereof, which can promote the growth of crops, improve the disease resistance of the crops, improve the absorption of the crops to nutrient substances, improve the utilization rate of pesticide and fertilizer and improve soil.
In order to achieve the aim, the invention provides a biostimulant of microbial origin, which comprises the following components in parts by weight: 1 to 50 percent of porphyridium, 1 to 50 percent of nitrogen-fixing blue algae, 1 to 50 percent of photosynthetic bacteria, 1 to 10 percent of carbon source and 1 to 10 percent of nitrogen source.
Preferably, the biostimulant of microbial origin comprises the following components in parts by weight: 10 to 35 percent of porphyridium, 10 to 35 percent of nitrogen-fixing blue algae, 5 to 25 percent of photosynthetic bacteria, 1 to 5 percent of carbon source and 1 to 5 percent of nitrogen source.
Preferably, in the microbial source biostimulation agent, the porphyridium is one of porphyridium liquid, porphyridium powder and porphyridium metabolic extract, and the porphyridium is porphyridium.
Preferably, in the microbial source biological stimulant, the nitrogen-fixing blue algae is one of nitrogen-fixing blue algae solution, nitrogen-fixing blue algae powder and nitrogen-fixing blue algae metabolic extract; the nitrogen-fixing blue algae species are as follows: one or more of Spirulina, anabaena, Nostoc, Gekko Swinhonis, Schistosoma, and Bifidobacterium.
Preferably, in the microbial source biological stimulator, the photosynthetic bacteria is one of a photosynthetic bacteria solution, a photosynthetic bacteria powder and a metabolic extract of the photosynthetic bacteria; the photosynthetic bacteria species are: one or more of rhodospirillum, cyanobacteria, protochlorella, rhodobacter, chlorobacter and rhodobacter sphaeroides.
Preferably, in the microbial source biostimulation agent, the carbon source is one or more of molasses powder, glucose, sucrose, starch, glycerol, sodium carbonate, sodium bicarbonate and sodium acetate;
the nitrogen source is one or more of urea, amino acid, peptone, ammonium chloride, ammonium nitrate, potassium nitrate and sodium nitrate.
Preferably, in the above microbial source biostimulation agent, the microbial source biostimulation agent further comprises an auxiliary material, the auxiliary material is one or more of a solvent, an adsorbent, a wetting agent, a dispersing agent and an emulsifying agent, and the microbial source biostimulation agent is a liquid preparation or a solid preparation.
Preferably, in the biostimulant of microbial origin, the biostimulant of microbial origin is a liquid preparation or a solid preparation.
Preferably, in the microbial source bio-stimulant, the solvent is one or more of water, methanol, ethanol, ethylene glycol, acetonitrile, xylene, epoxidized soybean oil, turpentine and nodel rosin vegetable oil.
Preferably, in the microbial source biostimulation agent, the adsorbent is one or more of polysaccharide, protein, anthracite, bamboo charcoal, activated carbon, white carbon black and glutaraldehyde glycidyl methacrylate copolymer.
Preferably, in the biostimulating agent of microbial origin, the dispersant is one or more of lignosulfonate, alkylnaphthalene formaldehyde condensate sulfonate, sulfate ester salt, methyl cellulose, alkylphenol polyoxyethylene, polyoxyethylene ether sulfonate and polycarboxylate.
Preferably, in the microbial source biostimulant, the emulsifier is one or more of calcium dodecylbenzene sulfonate, fatty alcohol-polyoxyethylene ether, alkyl glycoside, block polyether, alkylbenzene sulfonate, nonylphenol ether phosphate, and nonylphenol polyoxyethylene ether formaldehyde condensate sulfonate.
The preparation method of the microbial source biological stimulator comprises the following steps:
(1) preparing microalgae solution, microalgae powder or microalgae extract, wherein the microalgae solution comprises porphyridium solution and nitrogen-fixing cyanobacteria solution, the microalgae powder comprises porphyridium powder and nitrogen-fixing cyanobacteria powder, and the microalgae extract comprises porphyridium metabolic extract and nitrogen-fixing cyanobacteria metabolic extract;
the porphyridium algae liquid is prepared by the following steps: inoculating porphyridium into a culture medium A, and culturing to a growth stabilization period under the conditions that the culture temperature is 25-30 ℃, the light-dark time ratio is 16h:8h, and the illumination intensity of an LED light source is 2500-4000 Lux to obtain porphyridium liquid;
the preparation of the nitrogen-fixing blue algae solution comprises the following steps: inoculating nitrogen-fixing blue algae into a culture medium B, and culturing the nitrogen-fixing blue algae in a constant-temperature illumination bed until the growth stabilization period is reached under the conditions that the illumination intensity is 60000-80000 Lux, the light dark period is 12h:12h, the temperature is 28-30 ℃, the rotation speed of a shaking table is 120-200 r/min, and the pH is 7.0-7.5 to obtain nitrogen-fixing blue algae liquid;
(2) preparing photosynthetic bacteria liquid, photosynthetic bacteria powder or metabolic extract of photosynthetic bacteria;
the preparation of the photosynthetic bacteria liquid is as follows: inoculating photosynthetic bacteria into a basic culture medium, and culturing to a growth stabilization period under the conditions of illumination intensity of an LED light source of 3000-5000 Lux for 24 hours at the temperature of 28-30 ℃ in a constant-temperature illumination bed to obtain a photosynthetic bacteria liquid;
(3) uniformly mixing one of the microalgae solution, the microalgae powder and the microalgae extract with one of the photosynthetic bacteria liquid, the photosynthetic bacteria powder and the photosynthetic bacteria metabolic extract to obtain a mixture;
(4) and (4) adding a carbon source, a nitrogen source and auxiliary materials into the mixture obtained in the step (3) and uniformly mixing to obtain the biological stimulator.
Preferably, in the above method for producing a biostimulant derived from a microorganism, the medium a is: artificial seawater culture medium (ASW), KOCK culture medium, Harder-Bederke culture medium, Pringshein culture medium I, Pringshein culture medium II.
Preferably, in the method for producing a biostimulant of microbial origin, the culture medium is an artificial seawater culture medium (ASW).
Preferably, in the above method for preparing a microbial source biostimulating agent, the artificial seawater culture medium comprises a nitrogen source and a carbon source, and the nitrogen source is: one or more of urea, amino acid, ammonium chloride, ammonium nitrate, potassium nitrate and sodium nitrate, wherein the content of the nitrogen source is 0.5-2.0 g/L; the carbon source is: one or more of molasses powder, glucose, glycerol, sodium carbonate, sodium bicarbonate and sodium acetate, wherein the content of the carbon source is 1.0-20.0 g/L. The additional carbon source and nitrogen source are added into the culture medium, which is beneficial to improving the biomass of the porphyridium and producing more high value-added metabolites.
Preferably, in the preparation method of the microbial source biostimulation agent, in the step (1), the LED light source is green light, and the illumination intensity is 3000-3500 Lux.
Preferably, in the above method for producing a biostimulant of microbial origin, the medium B is: AA culture medium, BG11 culture medium, and aquatic 111 culture medium.
Preferably, in the above method for producing a biostimulant of microbial origin, the medium B is an AA medium.
Preferably, in the above method for preparing a biostimulating agent of microbial origin, in the step (2), the LED light source is: blue light or yellow light, and the illumination intensity is 4000-5000 Lux.
Preferably, in the above method for preparing a biostimulating agent derived from microorganisms, the preparation of the microalgal powder or the photosynthetic bacteria powder is: flocculating the microalgae solution or the photosynthetic bacteria solution cultured to the growth stabilization period by using a flocculating agent with the concentration of 0.01-0.5 g/L, filtering, and drying filter residues in vacuum to obtain microalgae powder or photosynthetic bacteria powder; the flocculating agent is as follows: one of aluminum sulfate, ferric chloride, polyaluminium sulfate, modified chitosan bentonite, modified chitosan diatomite and cationic starch.
Preferably, in the above method for preparing a biostimulant derived from microorganisms, the microalgae extract or the metabolic extract of photosynthetic bacteria is prepared by:
firstly, crushing microalgae or photosynthetic bacteria cells by adopting one of high pressure, ultrasonic wave, repeated freeze thawing, biological enzyme dissolution and ball milling methods;
② extracting water-soluble metabolites by one of alkali liquor extraction method, microwave-assisted extraction method and hot water extraction method, filtering and concentrating to powder I;
③ solvent extraction, enzymolysis and supercritical CO2Extracting water-insoluble metabolite by one of the extraction methods, filtering, and concentrating to obtain powder II;
and fourthly, mixing the powder I and the powder II to obtain the microalgae extract or the photosynthetic bacteria metabolic extract.
Compared with the prior art, the invention has the following beneficial effects:
1. the microbial source biological stimulator is compounded by microalgae and photosynthetic bacteria, has the effects of fertilizer and pesticide, can promote the metabolism and growth of crops, and improves the disease resistance of the crops.
2. The microbial source biological stimulator can effectively improve the absorption and stress resistance of nutrient substances by spraying on the leaf surfaces, can also improve the physical and chemical properties and community microorganisms of soil by spraying on the soil, and improves the yield and quality of crops.
3. In the microbial source biological stimulant, the nitrogen-fixing blue algae has a high nitrogen-fixing effect and can improve soil nutrients; the porphyridium produces high value-added metabolites such as phycobiliprotein, polyunsaturated fatty acid and extracellular polysaccharide substances, and can provide nutrient components for crops to promote the growth of the crops and improve the stress resistance; the photosynthetic bacteria have the capabilities of fixing nitrogen and dissolving phosphate, can provide nutrients for plants, can synthesize plant hormones, stimulate the growth of the plants and activate the immunity of the plants, and compared with the method of singly using one microorganism, the photosynthetic bacteria can effectively improve the nutrient absorption of crops, remarkably improve the utilization rate of pesticide and fertilizer, activate the stress resistance of the crops and improve the disease resistance of the crops by compounding three microorganisms and mutually synergizing three components.
Detailed Description
The following detailed description of specific embodiments of the invention is provided, but it should be understood that the scope of the invention is not limited to the specific embodiments.
Example 1
A microbial source biological stimulator is composed of the following components in parts by weight: 30% of porphyridium algae solution, 30% of spirulina solution, 20% of rhodococcus-like bacteria solution, 3% of urea, 3% of glucose and the balance of water.
The preparation method of the microbial source biological stimulator comprises the following steps:
(1) preparation of porphyridium algae solution: inoculating porphyridium into an artificial seawater culture medium (ASW), and culturing to a growth stabilization period under the conditions that the culture temperature is 25-30 ℃, the light-dark time ratio is 16h:8h, an LED green light source and the illumination intensity is 3500Lux to obtain porphyridium liquid; the ASW culture medium comprises a nitrogen source and a carbon source, wherein the nitrogen source is as follows: 1.0g/L of urea; the carbon source is glucose, and the content is 5.0 g/L;
(2) preparation of spirulina liquid: inoculating spirulina into an AA culture medium, and culturing to a growth stabilization period under the conditions of a constant-temperature illumination bed, the illumination intensity of 70000Lux, the light dark period of 12h:12h, the temperature of 28-30 ℃, the rotating speed of a shaking table of 150r/min and the pH of 7.0-7.5 to obtain spirulina liquid;
(3) the preparation of the rhodococcus-like bacterium liquid comprises the following steps: inoculating rhodococcus-like bacteria into a basic culture medium, and culturing the erythrococcus-like bacteria to a growth stabilization phase under the conditions of a constant-temperature illumination bed, an LED blue light source with illumination intensity of 4000Lux and illumination for 24 hours at the temperature of 28-30 ℃ to obtain a rhodococcus-like bacteria liquid;
(4) mixing the porphyridium algae solution, the spirulina algae solution and the rhodococcus-like bacteria solution, adding water, urea and glucose, and uniformly mixing to obtain the microbial source biological stimulator.
Example 2
A microbial source biological stimulator is composed of the following components in parts by weight: 25% of porphyridium algae liquid, 25% of spirulina liquid, 15% of rhodococcus-like bacteria liquid, 2% of ammonium nitrate, 2% of molasses powder, 3% of calcium lignosulfonate and the balance of activated carbon.
The method for preparing the microbial source biostimulant of the present example is different from that of example 1 in that: the ingredients were varied from the ingredient content and the other steps and parameters were the same as in example 1.
Example 3
A microbial source biological stimulator is composed of the following components in parts by weight: 25% of porphyridium powder, 20% of anabaena and spirulina mixed powder, 10% of rhodobacter sphaeroides powder, 3% of urea, 3% of sodium dodecyl benzene sulfonate, 5% of white carbon black and the balance of glucan.
The preparation method of the microbial source biological stimulator comprises the following steps:
(1) preparation of porphyridium algae powder: flocculating the porphyridium liquid cultured to the growth stabilization period by using a polyaluminium chloride flocculating agent with the concentration of 0.2g/L, filtering, and drying filter residues in vacuum to obtain microalgae powder;
(2) preparing mixed algae powder of anabaena and spirulina: flocculating the anabaena algae liquid and the spirulina liquid cultured to the growth stabilization period by using a modified chitosan bentonite flocculating agent with the concentration of 0.1g/L, filtering, drying filter residues in vacuum to obtain anabaena algae powder and spirulina powder, and then mixing the anabaena algae powder and the spirulina powder according to the weight ratio of 40:60 to obtain mixed algae powder of anabaena and spirulina;
(3) preparing rhodobacter violaceus powder: flocculating the rhodobacter sphaeroides liquid cultured to the growth stabilization phase by using cationic starch with the concentration of 0.2g/L, filtering, and drying filter residues in vacuum to obtain rhodobacter sphaeroides powder;
(4) mixing the porphyridium algae powder, anabaena and spirulina mixed algae powder, rhodobacter sphaeroides powder, urea, sodium dodecyl benzene sulfonate, white carbon black and glucan to obtain the microbial source biological stimulator.
Example 4
A microbial source biological stimulator is composed of the following components in parts by weight: 10% of porphyridium metabolic extract, 5% of schizandra metabolic extract, 5% of rhodospirillum metabolic extract, 2% of ethanol, 3% of alkyl glycoside and the balance of water.
The preparation method of the microbial source biological stimulator comprises the following steps:
(1) preparation of porphyridium metabolic extract, basket algae metabolic extract and rhodospirillum metabolic extract: crushing microalgae or photosynthetic bacteria cells by using ultrasonic waves; extracting water soluble metabolites by alkali liquor extraction, filtering and concentrating to powder I; extracting water-insoluble metabolites by a solvent extraction method, filtering and concentrating to obtain powder II; mixing the powder I and the powder II to obtain porphyridium metabolic extract, basket algae metabolic extract and rhodospirillum metabolic extract;
(2) mixing the metabolic extracts of porphyridium, Schistosoma and Rhodospirillum, adding ethanol, alkyl glycoside and water, and mixing to obtain the microbial source biostimulant.
Example 5
A microbial source biological stimulator is composed of the following components in parts by weight: 15 percent of porphyridium metabolic extract, 10 percent of anabaena metabolic extract, 10 percent of rhodospirillum metabolic extract, 3 percent of calcium dodecyl benzene sulfonate and the balance of white carbon black are mixed to prepare the biological stimulator.
Comparative example 1
The biostimulant of microbial origin of this comparative example differs from example 1: does not contain spirulina liquid and rhodococcus-like bacteria liquid, and the weight percentage of the porphyridium liquid is 80 percent.
Comparative example 2
The biostimulant of microbial origin of this comparative example differs from example 1: the spirulina liquid is 80 percent by weight except the porphyridium liquid and the rhodococcus-like bacteria liquid.
Comparative example 3
The biostimulant of microbial origin of this comparative example differs from example 1: not containing porphyridium algae liquid and spirulina algae liquid, the weight percentage of the rhodococcus-like bacteria liquid is 80%.
Comparative example 4
The biostimulant of microbial origin of this comparative example differs from example 1: not containing rhodococcus-like bacteria liquid, and the weight percentage of the porphyridium algae liquid and the spirulina algae liquid is 40 percent.
Comparative example 5
The biostimulant of microbial origin of this comparative example differs from example 1: the spirulina liquid does not contain porphyridium liquid, the weight percentage of the spirulina liquid is 48 percent, and the weight percentage of the rhodococcus-like bacteria liquid is 32 percent.
Comparative example 6
The biostimulant of microbial origin of this comparative example differs from example 1: does not contain spirulina liquid. The weight percentage of the porphyridium liquid is 48 percent, and the weight percentage of the rhodococcus-like bacteria liquid is 32 percent.
Comparative example 7
The biostimulant of microbial origin of this comparative example differs from example 4 in that: does not contain metabolic extracts of Schistosoma and Rhodospirillum, and the weight percentage of the metabolic extracts of porphyridium is 20%.
Comparative example 8
The biostimulant of microbial origin of this comparative example differs from example 4 in that: does not contain metabolic extracts of porphyridium and rhodospirillum, and the weight percentage of the metabolic extracts of the schizallium is 20 percent.
Comparative example 9
The biostimulant of microbial origin of this comparative example differs from example 4 in that: does not contain metabolic extracts of porphyridium and Schistosoma, and the weight percentage of the metabolic extracts of rhodospirillum is 20 percent.
Comparative example 10
The biostimulant of microbial origin of this comparative example differs from example 4 in that: does not contain the metabolic extract of rhodospirillum, the weight percentage of the metabolic extract of porphyridium is 13.3 percent, and the weight percentage of the metabolic extract of schinophyta is 6.7 percent.
Comparative example 11
The biostimulant of microbial origin of this comparative example differs from example 4 in that: does not contain porphyridium metabolic extract, the weight percentage of the schinophyta metabolic extract is 10 percent, and the weight percentage of the rhodospirillum metabolic extract is 10 percent.
Comparative example 12
The biostimulant of microbial origin of this comparative example differs from example 4 in that: does not contain any metabolic extract of Schistosoma. The weight percentage of the porphyridium metabolic extract is 13.3 percent, and the weight percentage of the rhodospirillum metabolic extract is 6.7 percent.
Test example 1
Tests were conducted to examine the influence of the microbial biostimulant prepared in example 1 and comparative examples 1 to 6 on the growth characteristics, sugar content and disease resistance of sugarcane.
The test is carried out at Jincheng Tun of Jiangzhou district of Chongxi left city in Guangxi 3 months in 2020, the variety of the sugarcane to be tested is the osmanthus sugar No. 55, the row spacing is 1.85 meters, and the seeding amount is 4500 effective buds per mu. The test is provided with 7 treatment groups and 1 control group, a random block design is adopted, each treatment group is repeated for 3 times, and the area of each cell is 1 mu. The microbial source biological stimulator dosage of each treatment group is 5L/mu, the microbial source biological stimulator is diluted by 500 times and then sprayed on soil once in the tillering stage and the jointing stage of the sugarcane, and the rest stages are managed according to a conventional mode; and (5) performing water and fertilizer management on the control group according to a conventional mode. The incidence of the sugarcane dead center seedlings, smut and tip rot of each treatment is investigated in 8 months; randomly selecting 3 points in each cell before sugarcane harvest, investigating plant height, effective stem number and stem diameter of each point 1, measuring yield and determining sugar content and yield during harvest. The test results are shown in tables 1 and 2.
As shown in Table 1, the microbial biostimulation agents of example 1 and comparative examples 1 to 6 can effectively reduce the incidence rates of the dead center seedlings, the smut and the tip rot, compared with the control group, the incidence rates of the dead center seedlings, the smut and the tip rot of the group of example 1 are respectively reduced by 37%, 53% and 46%, and meanwhile, the disease resistance effect of the group of example 1 is obviously better than that of the group of comparative examples 1 to 6, which shows that algae and bacteria in the microbial biostimulation agent of the invention have mutual synergistic action and effectively improve the disease resistance of sugarcane.
TABLE 1 sugarcane incidence in the treatment and control groups
| Group of | Treatment of | Seedling of dead heart | Smut disease | Tip rot disease |
| 1 | Control | 3.38％ | 2.26％ | 2.63％ |
| 2 | Example 1 | 2.12％ | 1.06％ | 1.41％ |
| 3 | Comparative example 1 | 2.54％ | 1.45％ | 2.17％ |
| 4 | Comparative example 2 | 2.97％ | 1.86％ | 1.86％ |
| 5 | Comparative example 3 | 2.50％ | 1.79％ | 2.14％ |
| 6 | Comparative example 4 | 2.47％ | 1.77％ | 1.77％ |
| 7 | Comparative example 5 | 2.45％ | 1.40％ | 1.75％ |
| 8 | Comparative example 6 | 2.49％ | 1.42％ | 1.78％ |
Table 2 shows the growth characteristics and sugar content of the sugar canes of each treatment group and the control group, and it can be seen from the table that the plant height, stem diameter, effective stem, sugar content and yield of the sugar canes are significantly improved by applying the microbial source biostimulant of example 1 and comparative examples 1 to 6 compared with the control group; the yield and the sugar increasing effect of the microbial source biostimulant of the example 1 containing the compound microorganism are better than those of the comparative examples 1 to 6.
TABLE 2 sugarcane growth traits and sugar content of the treatment groups and the control group
In conclusion, the algae and the bacteria in the microbial source biological stimulator have mutual synergistic effect, can effectively enhance the disease resistance of the sugarcane, promote the growth of the sugarcane, and have obvious yield and sugar increasing effect.
Test example 2
The weight-reducing and yield-increasing tests of the microbial biostimulants prepared in example 4 and comparative examples 7 to 12 were performed on tomatoes.
The tomato seeds are dispersed in the flowerpot with soil after germination acceleration, a layer of fine soil is uniformly covered, thinning is carried out after emergence of seedlings, and 3 seedlings are remained in each pot. The test is carried out by clear water contrast and 8 treatments: 2 grams of compound fertilizer is applied singly, 30 percent of compound fertilizer is reduced (1.4 grams) +0.5mL of example 4, 30 percent of compound fertilizer is reduced (1.4 grams) +0.5mL of comparative example 7, 30 percent of compound fertilizer is reduced (1.4 grams) +0.5mL of comparative example 8, 30 percent of compound fertilizer is reduced (1.4 grams) +0.5mL of comparative example 9, 30 percent of compound fertilizer is reduced (1.4 grams) +0.5mL of comparative example 10, 30 percent of compound fertilizer is reduced (1.4 grams) +0.5mL of comparative example 11, and 30 percent of compound fertilizer is reduced (1.4 grams) +0.5mL of comparative example 12. After each treatment is diluted by 500 times, the diluted solution is applied to roots in a seedling stage, a flowering stage, a fruit setting stage and a fruit expanding stage respectively, and the rest stages are managed according to a conventional mode. Each treatment was performed in 3 pots, 3 replicates were used, and the number of fruits per plant and the weight of fruits were investigated at harvest, and the test results are shown in Table 3.
TABLE 3 tomato yield for each treatment group
As can be seen from Table 3, compared with the clear water control, the application of each treatment to tomatoes can obviously improve the number of single plants and the weight and the yield of the single plants, and the tomato yield increasing effect is relatively good when the compound fertilizer is reduced by using the method of the invention of example 4 or comparative examples 7-12, the effect of reducing the compound fertilizer by 30% + the treatment of example 4 is the best, and the yield of the compound fertilizer reduced by 30% + the treatment group of example 4 is improved by 30.7% compared with the treatment group of the compound fertilizer. The microbial source biological stimulator composition has the effect of increasing the yield and can effectively reduce the dosage of chemical fertilizers.
The foregoing descriptions of specific exemplary embodiments of the present invention have been presented for purposes of illustration and description. It is not intended to limit the invention to the precise form disclosed, and obviously many modifications and variations are possible in light of the above teaching. The exemplary embodiments were chosen and described in order to explain certain principles of the invention and its practical application to enable one skilled in the art to make and use various exemplary embodiments of the invention and various alternatives and modifications as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the claims and their equivalents.
Claims (10)
1. A biostimulant of microbial origin, comprising the following components in parts by weight: 1 to 50 percent of porphyridium, 1 to 50 percent of nitrogen-fixing blue algae, 1 to 50 percent of photosynthetic bacteria, 1 to 10 percent of carbon source and 1 to 10 percent of nitrogen source.
2. The biostimulant of microbial origin according to claim 1, comprising the following components in weight fractions: 10 to 35 percent of porphyridium, 10 to 35 percent of nitrogen-fixing blue algae, 5 to 25 percent of photosynthetic bacteria, 1 to 5 percent of carbon source and 1 to 5 percent of nitrogen source.
3. The biostimulant of microbial origin according to claim 1, wherein the porphyridium is one of a porphyridium algal fluid, a porphyridium powder, a porphyridium metabolic extract, the porphyridium species being a porphyridium species; the nitrogen-fixing blue algae is one of nitrogen-fixing blue algae solution, nitrogen-fixing blue algae powder and nitrogen-fixing blue algae metabolic extract; the nitrogen-fixing blue algae species are as follows: one or more of Spirulina, anabaena, Nostoc, Gekko Swinhonis, Schistosoma, and Bifidobacterium.
4. The biostimulator of claim 1, wherein said photosynthetic bacteria is one of a photosynthetic bacteria broth, a photosynthetic bacteria powder, a metabolic extract of photosynthetic bacteria; the photosynthetic bacteria species are: one or more of rhodospirillum, cyanobacteria, protochlorella, rhodobacter, chlorobacter and rhodobacter sphaeroides.
5. The biostimulator of claim 1, wherein the carbon source is one or more of molasses, glucose, sucrose, starch, glycerol, sodium carbonate, sodium bicarbonate, sodium acetate; the nitrogen source is one or more of urea, amino acid, peptone, ammonium chloride, ammonium nitrate, potassium nitrate and sodium nitrate.
6. The biostimulation agent of claim 1, wherein said biostimulation agent further comprises adjuvants, and said adjuvants are one or more of solvent, adsorbent, wetting agent, dispersant, and emulsifier.
7. A method for preparing a biostimulant of microbial origin according to any of claims 1 to 6, comprising the steps of:
(1) preparing microalgae solution, microalgae powder or microalgae extract, wherein the microalgae solution comprises porphyridium solution and nitrogen-fixing cyanobacteria solution, the microalgae powder comprises porphyridium powder and nitrogen-fixing cyanobacteria powder, and the microalgae extract comprises porphyridium metabolic extract and nitrogen-fixing cyanobacteria metabolic extract;
the porphyridium algae liquid is prepared by the following steps: inoculating porphyridium into a culture medium A, and culturing to a growth stabilization period under the conditions that the culture temperature is 25-30 ℃, the light-dark time ratio is 16h:8h, and the illumination intensity of an LED light source is 2500-4000 Lux to obtain porphyridium liquid; the culture medium A is one of an artificial seawater culture medium, a KOCK culture medium, a Harder-Bederke culture medium and a Pringshein culture medium I, Pringshein culture medium II.
The preparation of the nitrogen-fixing blue algae solution comprises the following steps: inoculating nitrogen-fixing blue algae into a culture medium B, and culturing the nitrogen-fixing blue algae in a constant-temperature illumination bed until the growth stabilization period is reached under the conditions that the illumination intensity is 60000-80000 Lux, the light dark period is 12h:12h, the temperature is 28-30 ℃, the rotation speed of a shaking table is 120-200 r/min, and the pH is 7.0-7.5 to obtain nitrogen-fixing blue algae liquid; the culture medium B is one of an AA culture medium, a BG11 culture medium and an aquatic 111 culture medium;
(2) preparing photosynthetic bacteria liquid, photosynthetic bacteria powder or metabolic extract of photosynthetic bacteria;
the preparation of the photosynthetic bacteria liquid is as follows: inoculating photosynthetic bacteria into a basic culture medium, and culturing to a growth stabilization period under the conditions of illumination intensity of an LED light source of 3000-5000 Lux for 24 hours at the temperature of 28-30 ℃ in a constant-temperature illumination bed to obtain a photosynthetic bacteria liquid;
(3) uniformly mixing one of the microalgae solution, the microalgae powder and the microalgae extract with one of the photosynthetic bacteria liquid, the photosynthetic bacteria powder and the photosynthetic bacteria metabolic extract to obtain a mixture;
(4) and (4) adding a carbon source, a nitrogen source and auxiliary materials into the mixture obtained in the step (3) and uniformly mixing to obtain the biological stimulator.
8. The method for preparing a biostimulant of microbial origin according to claim 7, wherein in the step (1), the culture medium A is an artificial seawater culture medium, the artificial seawater culture medium further comprises a nitrogen source and a carbon source, and the nitrogen source is: one or more of urea, amino acid, ammonium chloride, ammonium nitrate, potassium nitrate and sodium nitrate, wherein the content of the nitrogen source is 0.5-2.0 g/L; the carbon source is: one or more of molasses powder, glucose, glycerol, sodium carbonate, sodium bicarbonate and sodium acetate, wherein the content of the carbon source is 1.0-20.0 g/L.
9. The method for preparing biostimulant of microbial origin according to claim 7, wherein the microalgal flour or photosynthetic bacteria flour is prepared by: flocculating the microalgae solution or the photosynthetic bacteria solution cultured to the growth stabilization period by using a flocculating agent with the concentration of 0.01-0.5 g/L, filtering, and drying filter residues in vacuum to obtain microalgae powder or photosynthetic bacteria powder; the flocculating agent is as follows: one of aluminum sulfate, ferric chloride, polyaluminium sulfate, modified chitosan bentonite, modified chitosan diatomite and cationic starch.
10. The method of claim 7, wherein the microalgae extract or the metabolic extract of photosynthetic bacteria is prepared by:
firstly, crushing microalgae or photosynthetic bacteria cells by adopting one of high pressure, ultrasonic wave, repeated freeze thawing, biological enzyme dissolution and ball milling methods;
② extracting water-soluble metabolites by one of alkali liquor extraction method, microwave-assisted extraction method and hot water extraction method, filtering and concentrating to powder I;
③ solvent extraction, enzymolysis and supercritical CO2Extracting water-insoluble metabolite by one of the extraction methods, filtering, and concentrating to obtain powder II;
and fourthly, mixing the powder I and the powder II to obtain the microalgae extract or the photosynthetic bacteria metabolic extract.
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