CN114437978A - Microbial compound inoculant for sugarcane planting and preparation method thereof - Google Patents
Microbial compound inoculant for sugarcane planting and preparation method thereof Download PDFInfo
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Abstract
The application relates to a microbial compound microbial inoculum for sugarcane planting and a preparation method thereof, wherein the microbial compound microbial inoculum comprises the following components in percentage by mass: 35-40wt% of compound microbial inoculum A; 50-55wt% of compound microbial inoculum B; 5-15wt% of a regulator; the compound microbial inoculum A comprises 55-60wt% of bacillus licheniformis suspension, 25-30wt% of cercospora suspension and 10-20wt% of alginic acid; the composite microbial inoculum B at least comprises 45-50wt% of bacillus mucilaginosus bacterial suspension, 25-30wt% of photosynthetic bacteria bacterial suspension and 16-30wt% of silicate bacteria bacterial suspension. The application has the effect of improving the yield and the sugar content of the sugarcane.
Description
Technical Field
The application relates to the field of microbial compound inocula, in particular to a microbial compound inocula for sugarcane planting and a preparation method thereof.
Background
Sugar cane is a temperate and tropical crop, sucrose is one of the end products of sugar cane photosynthesis, and is also the primary form of photosynthetic product transport and distribution within most plants. "double high" sugarcane means sugarcane that is high in yield and sugar content. The sugarcane is used as a raw material for preparing cane sugar and extracting ethanol, has higher economic benefit and is widely planted and popularized in southern areas of China.
Sugarcane is a raw material of the sugar industry, and sugarcane sugar is a source of economic benefits of the sugar industry. The economic benefit of sugar industry means that the three main factors of the amount of sugar cane pressed, the sugar yield and the cost of sugar per ton are mutually linked, and the total profit of sugar products and the profit of ten thousand tons of sugar cane are comprehensively reflected by the financial accounting in a complementary way. The sugarcane squeezing amount is a basis and is related to the profit of ten thousand tons of sugarcane, but the sugarcane has high sucrose content and does not have a certain amount of raw sugarcane, so that the sugarcane squeezing period is short, the sugarcane yield is low, the equipment utilization rate is low, and the economic benefit is not high. However, although a certain amount of raw sugarcane is available, the sugar content of the raw sugarcane is low, the sugar yield is low, the cost is high, and the economic benefit is not great. Only the raw material cane with sufficient quantity and high quality can obtain the maximum economic benefit.
Aiming at the technical problems, the inventor thinks that the double-high sugarcane with high yield and high sugar content is obtained by regulating and improving the yield and the sugar content of the sugarcane through the life metabolic activity of microorganisms in the microbial fertilizer, and is the direction of key research in the sugarcane planting in the future.
Disclosure of Invention
In order to improve the yield and sugar content of sugarcane, the application provides a microbial compound inoculant for sugarcane planting and a preparation method thereof.
In a first aspect, the application provides a microbial composite inoculant for sugarcane planting, which adopts the following technical scheme:
a microbial compound microbial inoculant for sugarcane planting comprises the following components in percentage by mass:
35-40wt% of compound microbial inoculum A;
50-55wt% of compound microbial inoculum B;
5-15wt% of a regulator;
the complex microbial inoculum A comprises 55-60wt% of bacillus licheniformis suspension, 25-30wt% of cercospora suspension and 10-20wt% of alginic acid;
the preparation method of the complex microbial inoculum A comprises the following steps:
s1-1, respectively performing solid activation culture on the Bacillus licheniformis and Cercospora species to obtain the final product with concentration of 1-3 × 108CFU/ml of Bacillus licheniformis suspension liquid and Cercospora species suspension liquid;
s1-2, inoculating the bacillus licheniformis suspension liquid to a PDA culture medium in an inoculation amount of 5-7%, performing single-plant shake flask fermentation culture on a shaking table at the culture temperature of 32-35 ℃, the shaking speed of the shaking table of 150-170rpm, and the culture time of 72-76h to obtain the bacillus licheniformis suspension;
s1-3, inoculating the cercospora suspension liquid to a PDA culture medium by 3-5% of inoculum size, carrying out single-plant shake flask fermentation culture on a shaking table, wherein the culture temperature is 32-35 ℃, the shaking rate of the shaking table is 150-170rpm, and the culture time is 72-76h, so as to obtain the cercospora suspension;
s1-4, mixing the bacillus licheniformis suspension, the cercospora suspension and alginic acid, adding into a stirring pot, and stirring at a constant speed of 100r/min for 15-20min to obtain a composite microbial inoculum A;
the complex microbial inoculum B at least comprises 45-50wt% of Bacillus mucilaginosus bacterial suspension, 25-30wt% of photosynthetic bacteria bacterial suspension and 16-30wt% of silicate bacteria bacterial suspension;
the preparation method of the compound microbial inoculum B comprises the following steps:
s2-1, respectively performing solid activation culture on Bacillus mucilaginosus strain, photosynthetic bacteria strain and silicate bacteria strain to obtain the culture solution with concentration of 1-3 × 108CFU/ml of Bacillus mucilaginosus suspension liquid, photosynthetic bacteria suspension liquid and silicate bacteria suspension liquid;
s2-2, inoculating the Bacillus mucilaginosus bacterial suspension liquid to a PDA culture medium in an inoculation amount of 5-7%, performing single-plant shake flask fermentation culture on a shaking table at the culture temperature of 32-35 ℃, the shaking table oscillation speed of 150-170rpm, and the culture time of 72-76h to obtain the Bacillus mucilaginosus bacterial suspension;
s2-3, inoculating the photosynthetic bacteria suspension liquid to an RCVBN culture medium by 3-5% of inoculation amount, carrying out single plant shake flask fermentation culture on a shaking table, wherein the culture temperature is 32-35 ℃, the shaking table oscillation speed is 150-170rpm, and the culture time is 72-76h, so as to obtain photosynthetic bacteria suspension;
s2-4, inoculating the silicate bacteria suspension liquid to an HB8548-2 culture medium in an inoculation amount of 2-4%, carrying out single-plant shake flask fermentation culture on a shaking table at the culture temperature of 30-32 ℃, the shaking table oscillation speed of 200-220rpm, and the culture time of 48-50h to obtain the silicate bacteria suspension;
s2-5, mixing the colloidal bacillus bacteria suspension, the photosynthetic bacteria suspension and the silicate bacteria suspension, adding into a stirring pot, and stirring at a constant speed of 100r/min for 15-20min to obtain the composite microbial agent B.
Preferably, the complex microbial inoculum A comprises the following components in percentage by mass: 57-58wt% of bacillus licheniformis suspension, 27-28wt% of cercospora suspension and 14-16wt% of alginic acid suspension; the composite microbial inoculum B at least comprises the following components in percentage by mass: 47-50wt% of Bacillus mucilaginosus bacterial suspension, 25-28wt% of photosynthetic bacteria bacterial suspension and 16-22wt% of silicate bacteria bacterial suspension.
Preferably, the complex microbial inoculum A comprises the following components in percentage by mass: 57.5wt% of a Bacillus licheniformis suspension, 27.5wt% of a Cercospora suspension and 15wt% of alginic acid; the composite microbial inoculum B at least comprises the following components in percentage by mass: 48.5wt% Bacillus mucilaginosus bacterial suspension, 26.5wt% photosynthetic bacteria bacterial suspension and 19wt% silicate bacteria bacterial suspension.
The sugar accumulation of sugarcane is a complex process which is regulated and controlled by a plurality of sucrose metabolic enzymes and relates to physiological processes of sucrose synthesis, decomposition, transportation and the like. The enzyme can catalyze sucrose synthesis and regulate sucrose content. Among them, Sucrose Phosphate Synthase (SPS), Sucrose Synthase (SS) and Invertase (INV) are 3 major key enzymes for regulating sucrose metabolism. SPS is responsible for the synthesis of sucrose; SS is a reversible enzyme which can catalyze the synthesis of sucrose and the decomposition of sucrose; INV plays an important role in the transport, storage and distribution of sucrose. In plants, SPS, expressed both in the source and sink, is involved in sucrose resynthesis, regulating starch accumulation, protein storage and cellulose synthesis.
By adopting the technical scheme, the combined action of the bacillus licheniformis, the cercospora and the alginic acid in the composite microbial agent A can enhance the activity of amylase, protease and Invertase (INV), promote the decomposition and transformation of sucrose and nutrient substances, make the nutrient substances easier to be absorbed by plants and help to promote the plant growth of sugarcane; the bacillus mucilaginosus, the photosynthetic bacteria and the silicate bacteria in the composite microbial agent B act together, so that the generation and conversion of various elements such as nitrogen, potassium, phosphorus, boron and the like in the soil are increased, and the sugarcane sugar content accumulation is enhanced; the regulator is mainly plant growth hormone; the addition of the regulator can stimulate metabolic enzymes in the sugarcane to a certain extent, and the regulator, the compound microbial inoculum A and the compound microbial inoculum B are mixed according to a certain proportion and act together, so that the accumulation or conversion of cane sugar in the sugarcane is favorably balanced, the dynamic balance of high-speed conversion of cane sugar is realized, the growth of the sugarcane and the accumulation of cane sugar are further promoted, and the aims of high yield and high sugar content of the sugarcane are further realized. The components in the three components act together, and the inhibition and resistance effects of the sugarcane on the external fungi and bacteria are improved.
Preferably, the regulator is one or more of fulvic acid, glyphosate and gibberellin.
By adopting the technical scheme, the fulvic acid can promote the growth of plants, particularly can properly control the opening degree of leaf stomata of crops, reduce transpiration, improve the stress resistance of the plants and improve the quality; meanwhile, the activity of the plant root system can be improved, and various enzyme activities can be improved. Glyphosate mainly inhibits enolpyruvylshikimate phosphate synthase in plants, thereby inhibiting the conversion of shikimate to phenylalanine, tyrosine and tryptophan and interfering protein synthesis. Gibberellin can accelerate cell elongation and promote cell division. The balance of the compound microbial inoculum A and the compound microbial inoculum B on the synthesis and decomposition of the cane sugar can be effectively regulated and controlled by single or mixed use of the three, so that the contradiction between insufficient accumulation of the cane sugar or insufficient conversion of the cane sugar to hinder the growth due to too fast growth is relieved, the growth of the cane sugar is good, and the content of the cane sugar is also improved.
Preferably, the regulator is prepared by mixing fulvic acid, glyphosate and gibberellin according to the mass ratio of (0.5-0.8) to (1-1.1) to (1.2-1.5).
By adopting the technical scheme, the fulvic acid, the glyphosate and the gibberellin are mixed and used in a specific ratio, and the fulvic acid, the glyphosate and the gibberellin interact with each other, so that the composite microbial inoculum A, the composite microbial inoculum B and the regulator can be fully combined, the decomposition and accumulation of the cane sugar are in a balanced state, and the yield and the sugar of the cultivated cane sugar are improved.
Preferably, the complex microbial inoculum B also comprises the following components in percentage by mass: 3-5wt% azotobacter suspension, 2-4wt% phosphate solubilizing bacteria suspension and 3-5wt% potassium solubilizing bacteria suspension;
the preparation method of the azotobacter bacterial suspension comprises the following steps: activating and culturing azotobacteria strain to obtain the product with concentration of 1-3 × 108CFU/ml azotobacter suspension liquid; inoculating the azotobacter suspension liquid to No. 77 azotobacter culture medium with the inoculation amount of 1-3%, performing single-strain shake flask fermentation culture on a shaking table at the culture temperature of 28-30 ℃, the shaking table oscillation speed of 200-220rpm, and the culture time of 48-50h to obtain azotobacter suspension;
the preparation method of the phosphate solubilizing bacteria suspension comprises the following steps: activating and culturing solid of phosphorus-solubilizing bacteria strain to obtain the product with concentration of 1-3 × 108CFU/ml of phosphate solubilizing bacteria suspension liquid; inoculating the phosphate solubilizing bacteria suspension liquid to an HB8549-2 phosphate solubilizing bacteria culture medium by an inoculation amount of 1-3%, performing single-strain shake flask fermentation culture on a shaking table at the culture temperature of 30-32 ℃, the shaking table oscillation speed of 200-220rpm, and the culture time of 48-50h to obtain the phosphate solubilizing bacteria suspension;
the preparation method of the potassium bacteria dissolving suspension comprises the following steps: activating and culturing solid potassium-decomposing bacteria strain to obtain the product with concentration of 1-3 × 108CFU/ml of potassium bacteria suspension liquid; inoculating the suspension liquid of the potassium bacteria to a potassium solution culture medium by the inoculation amount of 1-3%, and performing single-plant shake flask fermentation culture on a shaking table at the culture temperature of 37-40 ℃, the shaking table oscillation speed of 200-220rpm and the culture time of 48-50h to obtain the suspension of the potassium bacteria.
By adopting the technical scheme, the three substances are added into the composite microbial agent B, and the three microorganisms can stably and quickly accumulate nutrient substances such as nitrogen, phosphorus, potassium and the like in soil, so that the nutrition of the nutrient substances to the sugarcane is further promoted, and the sugar content of the sugarcane is promoted to be increased.
Preferably, the azotobacter strain is azospirillum strain or paspalum azotobacter strain.
By adopting the technical scheme, the azospirillum and paspalum azotobacter can live between cortical cells in sugarcane roots. The two have certain specificity with symbiotic sugarcane, play a role in combined nitrogen fixation, and contribute to further improving the nitrogen fixation effect on the sugarcane.
In a second aspect, the preparation method of the microbial compound inoculant for sugarcane planting provided by the application adopts the following technical scheme:
a preparation method of a microbial compound inoculant for sugarcane planting comprises the following steps: and (3) uniformly mixing the compound microbial inoculum A, the compound microbial inoculum B and the regulator, and fermenting to obtain the microbial compound microbial inoculum.
By adopting the technical scheme, the composite microbial inoculum A and the composite microbial inoculum B are prepared respectively, so that the components in the composite microbial inoculum A and the composite microbial inoculum B can be uniformly mixed, the combined action of the components in the composite microbial inoculum A and the composite microbial inoculum B is facilitated, and the respective effects of the composite microbial inoculum A and the composite microbial inoculum B are improved; and finally, uniformly mixing the regulator with the composite microbial inoculum A and the composite microbial inoculum B, so that a microbial composite microbial inoculum system is balanced and stable, and the yield and sugar content of sugarcane applied with the microbial composite microbial inoculum are improved.
In summary, the present application includes at least one of the following beneficial technical effects:
1. the composite microbial inoculum A in the application has the combined action of bacillus licheniformis, cercospora and alginic acid, and is beneficial to promoting the growth of sugarcane plants; the bacillus mucilaginosus, the photosynthetic bacteria and the silicate bacteria contained in the compound microbial inoculum B act together, so that the generation and the stability of various elements such as nitrogen, potassium, phosphorus, boron and the like in soil are facilitated, and the accumulation of the content of sucrose in sugarcane can be promoted; the regulator is mainly plant growth hormone; the regulator, the complex microbial inoculum A and the complex microbial inoculum B are mixed and act together, so that the accumulation or transformation of cane sugar in the cane can be better regulated and controlled, and the aims of high yield and high sugar content of the cane can be further realized;
2. the regulator is prepared by mixing fulvic acid, glyphosate and gibberellin according to the mass ratio of (0.5-0.8) to (1-1.1) to (1.2-1.5); the regulator prepared by mixing the three components has better compatibility with the compound microbial inoculum A and the compound microbial inoculum B, is beneficial to balancing the influence of the compound microbial inoculum A and the compound microbial inoculum B on the growth process of the sugarcane, and further promotes the yield and the sugar content of the sugarcane;
3. the composite microbial inoculum B also comprises azotobacter, phosphate solubilizing bacteria and potassium solubilizing bacteria, and the addition of the three beneficial bacteria can quickly and stably accumulate nutrient substances such as nitrogen, phosphorus, potassium and the like in soil, so that the growth of sugarcane and the accumulation of sucrose are further promoted.
Detailed Description
The present application will be described in further detail with reference to examples.
The species of Bacillus licheniformis, Cercospora species, Bacillus mucilaginosus, photosynthetic bacteria, azotobacter species, phosphate solubilizing species, potassium solubilizing species and silicate bacteria, and the media required accordingly, as required in the examples and comparative examples of the present application, were commercially available.
Examples
Example 1
The embodiment discloses a microbial compound inoculant for sugarcane planting, which comprises the following raw materials:
3.5kg of compound microbial inoculum A; 5.0kg of compound microbial inoculum B; 1.5kg of conditioning agent; the regulator is ethephon.
The compound microbial inoculum A comprises 55wt% of bacillus licheniformis suspension, 25 wt% of cercospora suspension and 20wt% of alginic acid; the preparation method of the compound microbial inoculum A comprises the following steps:
s1-1, respectively performing solid activation culture on the Bacillus licheniformis and Cercospora species to obtain the final product with concentration of 1-3 × 108CFU/ml of Bacillus licheniformis suspension liquid and Cercospora species suspension liquid;
s1-2, inoculating the bacillus licheniformis suspension liquid to a PDA culture medium in an inoculation amount of 5%, carrying out single-plant shake flask fermentation culture on a shaking table at the culture temperature of 32 ℃, the shaking table oscillation speed of 150rpm, and the culture time of 72h to obtain bacillus licheniformis suspension;
s1-3, inoculating the cercospora suspension liquid to a PDA culture medium in an inoculation amount of 3%, carrying out single-strain shake flask fermentation culture on a shaking table at the culture temperature of 32 ℃, the shaking table oscillation speed of 150rpm, and the culture time of 72h to obtain the cercospora suspension;
s1-4, mixing 5.5kg of bacillus licheniformis suspension, 2.5kg of cercospora suspension and 2kg of alginic acid, adding into a stirring pot, and stirring at a constant speed of 100r/min for 15min to obtain a composite microbial inoculum A;
the compound microbial inoculum B comprises 45 wt% of bacillus mucilaginosus suspension, 25 wt% of photosynthetic bacteria suspension and 30wt% of silicate bacteria suspension. The preparation method of the compound microbial inoculum B comprises the following steps:
s2-1, respectively performing solid activation culture on Bacillus mucilaginosus strain, photosynthetic bacteria strain and silicate bacteria strain to obtain the culture solution with concentration of 1-3 × 108CFU/ml of Bacillus mucilaginosus suspension liquid, photosynthetic bacteria suspension liquid and silicate bacteria suspension liquid;
s2-2, inoculating the bacillus mucilaginosus suspension liquid to a PDA culture medium in an inoculation amount of 5%, carrying out single-strain shake flask fermentation culture on a shaking table at the culture temperature of 35 ℃, the shaking table oscillation speed of 170rpm, and the culture time of 73h to obtain the bacillus mucilaginosus suspension;
s2-3, inoculating the photosynthetic bacteria suspension liquid to an RCVBN culture medium with an inoculation amount of 5%, carrying out single plant shake flask fermentation culture on a shaking table at a culture temperature of 35 ℃, a shaking table oscillation speed of 170rpm, and a culture time of 73h to obtain photosynthetic bacteria suspension;
s2-4, inoculating the silicate bacteria suspension liquid to an HB8548-2 culture medium in an inoculation amount of 4%, carrying out single strain shake flask fermentation culture on a shaking table, wherein the culture temperature is 32 ℃, the shaking table oscillation speed is 220rpm, and the culture time is 50h, so as to obtain silicate bacteria suspension;
s2-5, mixing 4.5kg of colloidal bacillus bacterial suspension, 2.5kg of photosynthetic bacteria bacterial suspension and 3kg of silicate bacteria bacterial suspension, adding into a stirring pot, and stirring at a constant speed of 100r/min for 20min to obtain the compound microbial inoculum B.
The embodiment also discloses a preparation method of the microbial compound inoculant for sugarcane planting, which comprises the following steps: adding 3.5kg of compound microbial inoculum A, 5kg of compound microbial inoculum B and 1.5kg of regulator into a reaction kettle, stirring once every 5min under the conditions that the heating temperature is 35 ℃ and the stirring speed is 500r/min, taking out after heating and stirring for 30min, standing and cooling for 1h, fermenting until the concentration is 2 x 1012CFU/ml and the number of spores accounts for more than 85 percent of the total number of bacteria in the zymogen liquid, and the microbial composite inoculum is obtained.
Example 2
The embodiment discloses a microbial compound inoculant for sugarcane planting, which comprises the following raw materials:
4.0kg of compound microbial inoculum A; 5.5kg of compound microbial inoculum B; 0.5kg of conditioning agent; the regulator is ethephon.
The preparation method of the compound microbial inoculum A comprises the following steps:
s1-1, respectively performing solid activation culture on the Bacillus licheniformis and Cercospora species to obtain the final product with concentration of 1-3 × 108CFU/ml of Bacillus licheniformis suspension liquid and Cercospora species suspension liquid;
s1-2, inoculating the bacillus licheniformis suspension liquid to a PDA culture medium in an inoculation amount of 7%, performing single-strain shake-flask fermentation culture on a shaking table at the culture temperature of 35 ℃, the shaking table oscillation speed of 170rpm, and the culture time of 73h to obtain the bacillus licheniformis suspension;
s1-3, inoculating the cercospora suspension liquid to a PDA culture medium in an inoculation amount of 5%, carrying out single-strain shake-flask fermentation culture on a shaking table at the culture temperature of 35 ℃, the shaking table oscillation speed of 170rpm, and the culture time of 73h to obtain the cercospora suspension;
s1-4, mixing 5.5kg of bacillus licheniformis suspension, 2.5kg of cercospora suspension and 2kg of alginic acid, adding into a stirring pot, and stirring at a constant speed of 100r/min for 20min to obtain a composite microbial inoculum A;
the preparation method of the compound microbial inoculum B comprises the following steps:
s2-1, mixing the colloidRespectively performing solid activation culture on bacillus strain, photosynthetic bacteria strain and silicate bacteria strain to obtain the product with concentration of 1-3 × 108CFU/ml of Bacillus mucilaginosus suspension liquid, photosynthetic bacteria suspension liquid and silicate bacteria suspension liquid;
s2-2, inoculating the bacillus mucilaginosus suspension liquid to a PDA culture medium in an inoculation amount of 5%, carrying out single-strain shake flask fermentation culture on a shaking table at the culture temperature of 32 ℃, the shaking table oscillation speed of 150rpm, and the culture time of 72h to obtain the bacillus mucilaginosus suspension;
s2-3, inoculating the photosynthetic bacteria suspension liquid to an RCVBN culture medium with the inoculation amount of 3%, carrying out single plant shake flask fermentation culture on a shaking table, wherein the culture temperature is 32 ℃, the shaking table oscillation speed is 150rpm, and the culture time is 72h, so as to obtain photosynthetic bacteria suspension;
s2-4, inoculating the silicate bacteria suspension liquid to an HB8548-2 culture medium in an inoculation amount of 2%, carrying out single strain shake flask fermentation culture on a shaking table, wherein the culture temperature is 30 ℃, the shaking speed of the shaking table is 200rpm, and the culture time is 48h, so as to obtain silicate bacteria suspension;
s2-5, mixing 4.5kg of colloidal bacillus bacterial suspension, 2.5kg of photosynthetic bacteria bacterial suspension and 3kg of silicate bacteria bacterial suspension, adding into a stirring pot, and stirring at a constant speed of 100r/min for 20min to obtain the compound microbial inoculum B.
The embodiment also discloses a preparation method of the microbial compound inoculant for sugarcane planting, which comprises the following steps:
adding 4kg of compound microbial inoculum A, 5.5kg of compound microbial inoculum B and 0.5kg of regulator into a reaction kettle, stirring once every 5min under the conditions that the heating temperature is 37 ℃ and the stirring speed is 450r/min, taking out after heating and stirring for 35min, standing and cooling for 1h, fermenting until the concentration is 2 multiplied by 1012CFU/ml and the number of spores accounts for more than 85 percent of the total number of bacteria in the zymogen liquid, and the microbial composite inoculum is obtained.
Examples 3 to 6
The embodiment is different from the embodiment 1 in that: the content of each bacterial suspension in the compound bacterial agent A, B is different and the material for the regulator is different. See table 1 for details.
TABLE 1
Example 7
The embodiment is different from the embodiment 1 in that: the regulator is prepared by mixing 0.28kg of fulvic acid, 0.56kg of glyphosate and 0.66kg of gibberellin, wherein the mass ratio of the fulvic acid to the glyphosate to the gibberellin is 0.5: 1: 1.2.
example 8
The embodiment is different from the embodiment 1 in that: the regulator is prepared by mixing 0.35kg of fulvic acid, 0.49kg of glyphosate and 0.66kg of gibberellin, wherein the mass ratio of the fulvic acid to the glyphosate to the gibberellin is 0.8: 1.1: 1.5 mixing the obtained regulator.
Example 9
The embodiment is different from the embodiment 1 in that: azotobacter strain, phosphate solubilizing bacterium strain and potassium solubilizing bacterium strain are also inoculated in the step 2-1, and in the step 2-2, 0.3kg azotobacter suspension, 0.2kg phosphate solubilizing bacterium suspension and 0.3kg potassium solubilizing bacterium suspension are mixed with 4.5kg colloidal bacillus bacterium suspension, 2.5kg photosynthetic bacteria suspension and 2.2kg silicate bacteria suspension and added into a stirring pot; wherein the nitrogen-fixing strain is rhizobium.
Example 10
The embodiment is different from the embodiment 1 in that: azotobacter strain, phosphate solubilizing bacterium strain and potassium solubilizing bacterium strain are also inoculated in the step 2-1, and in the step 2-2, 0.5kg azotobacter suspension, 0.4kg phosphate solubilizing bacterium suspension and 0.5kg potassium solubilizing bacterium suspension are mixed with 4.5kg colloidal bacillus bacterium suspension, 2.5kg photosynthetic bacteria suspension and 1.6kg silicate bacteria suspension and added into a stirring pot; wherein the nitrogen-fixing strain is azospirillum.
Example 11
The embodiment is different from the embodiment 10 in that: the nitrogen-fixing strain is selected from paspalum nitrogen-fixing bacteria.
Example 12
The present embodiment is different from embodiment 10 in that: the regulator is prepared by mixing 0.28kg of fulvic acid, 0.56kg of glyphosate and 0.66kg of gibberellin, wherein the mass ratio of the fulvic acid to the glyphosate to the gibberellin is 0.5: 1: 1.2.
comparative example
Comparative example 1
A microbial compound inoculant for sugarcane planting is different from that in example 1 in that: does not contain compound bacteria B and regulator.
Comparative example 2
A microbial compound inoculant for sugarcane planting is different from that in example 1 in that: does not contain the complex microbial inoculum A and a regulator.
Comparative example 3
A microbial compound inoculant for sugarcane planting is different from that in example 1 in that: does not contain compound bacteria agent A and compound bacteria agent B.
Comparative example 4
A microbial compound inoculant for sugarcane planting is different from that in example 1 in that: no regulator is included.
Comparative example 5
A microbial compound inoculant for sugarcane planting is different from that in example 1 in that: does not contain the compound bacterial agent B.
Comparative example 6
A microbial compound inoculant for sugarcane planting is different from that in example 1 in that: does not contain the complex microbial inoculum A.
Comparative example 7
A microbial compound inoculant for sugarcane planting is different from that in example 1 in that: replacing the bacillus licheniformis suspension with the equivalent bacillus amyloliquefaciens suspension.
Comparative example 8
A microbial compound inoculant for sugarcane planting is different from that in example 1 in that: the cercospora suspension is replaced by the same amount of actinomycetes suspension, the preparation method of the actinomycetes suspension is the same as that of the cercospora suspension, and the culture medium is a PDA (potato dextrose agar) culture medium.
Comparative example 9
A microbial compound inoculant for sugarcane planting is different from that in example 1 in that: alginic acid was replaced with equal amount of fulvic acid.
Performance test and data analysis
Test No.)
(1) Planting time: year 2020, 4-12 months
(2) Planting place: the pond area is the camphorwood town Deng Bao village and Huanghe village;
(3) the test varieties are: gui Tang No. 44
(4) Scheme design:
1. dividing the test field in Huanghe village into equal 22 parts, and recording as 1-1, 1-2, 1-3, etc.; planting 10 new plants of osmanthus sugar No. 44 in each part;
2. selecting 22 equal test fields in Deng Bao villages, and recording as 2-1, 2-2, 2-3 and the like; planting 10 plants of pachyrhizus 44 perennial root in each part;
(5) sample preparation: a commercial compound fertilizer of 125kg plus 2kg of microbial compound inoculum (one part of the Deng Bamura test field and the Huanghe village test field corresponds to the microbial compound inoculum prepared in one embodiment or the comparative example) is selected, and the other part does not use the microbial compound inoculum as a blank control group.
(6) As a result: the data of the test tests of examples 1 to 12 and comparative examples 1 to 9 and of the blank control are detailed in Table 2 (data of each test field are averaged over 10 sugarcane plants).
TABLE 2
According to the performance detection data of the sugarcane in the two test fields of the newly planted sugarcane and the perennial root in the table 2, the sugar content and the growth activity of the newly planted sugarcane are more excellent than those of the perennial root.
The microbial compound inoculant prepared in the embodiments 1 to 12 comprises three components, namely a compound inoculant A, a compound inoculant B and a regulator, and the three components are mutually promoted and act together, so that the sugarcane using the compound inoculant prepared in the embodiments 1 to 12 has better sugarcane yield and higher sugar content compared with a blank control group, and is closer to the planting target of 'double-high' sugarcane.
The sugarcane obtained by using the composite microbial inoculum prepared in the comparative example 1 is improved in plant height, stem diameter, sucrose content and field weight compared with the sugarcane obtained by a blank control, particularly the plant height and stem diameter are obviously improved, and the composite microbial inoculum A has the effect of promoting the growth of the sugarcane; the comparative example 2 does not contain the compound microbial inoculum A and the regulator, the compound microbial inoculum B is additionally added in the comparative example 2 compared with a blank control group, and the plant height, the stem diameter, the sucrose content and the field brix of the sugarcane obtained by using the compound microbial inoculum prepared in the comparative example 2 are improved in comparison with the sugarcane obtained by using the blank control group, particularly the sucrose content and the field brix are obviously improved, so that the compound microbial inoculum B has the effect of improving the sucrose content of the sugarcane.
The sugarcane obtained by using the composite microbial inoculum prepared in the comparative example 4 has smaller plant height, stem diameter, sucrose content and field brix than the sugarcane obtained in the comparative examples 1 and 2, and the condition that the composite microbial inoculum A and the composite microbial inoculum B possibly generate mutual inhibition when used together is shown; the comparative example 5 is the mixture of the complex microbial inoculum A and the regulator, the comparative example 6 is the mixture of the complex microbial inoculum B and the regulator, the performance detection data values of the comparative examples 5 and 6 are also higher than the performance detection data value of the comparative example 4, and the inhibition effect between the complex microbial inoculum A and the complex microbial inoculum B is further reflected.
The inventor analyzes that: the bacillus licheniformis, the cercospora and the alginic acid in the composite microbial agent A act together, so that the metabolism of the sugarcane is promoted, and the growth efficiency of plants is improved; the bacillus mucilaginosus, the photosynthetic bacteria and the silicate bacteria in the composite microbial agent B act together, so that the generation and stability of various elements such as nitrogen, potassium, phosphorus, boron and the like in the soil are increased, and the accumulation of the content of cane sugar in the sugarcane is facilitated; however, as the growth of plants needs to decompose part of the accumulated sucrose into nutrient substances required by growth, one of the compound microbial inoculum A and the compound microbial inoculum B synthesizes the sucrose, and the other promotes the decomposition and consumption of the sucrose, so that the mutual inhibition between the compound microbial inoculum A and the compound microbial inoculum B can be generated, and the mutual inhibition between the compound microbial inoculum A and the compound microbial inoculum is mainly added into a regulator of plant growth hormone; the regulator, the compound microbial inoculum A and the compound microbial inoculum B are mixed and act together, so that the accumulation or transformation of cane sugar in a system tends to be balanced, the plant growth and the accumulation of cane sugar are further promoted, and the effects of high yield and high sugar content of sugarcane are further achieved.
In the comparative example 7, the bacillus licheniformis is replaced by the same amount of bacillus amyloliquefaciens, and the gain effect of the composite microbial inoculum prepared in the comparative example 7 on the plant height and the stem diameter of the sugarcane is weakened, so that the mixing of the bacillus amyloliquefaciens, the cercospora and the alginic acid can not achieve the gain effect of the bacillus licheniformis, the cercospora and the alginic acid on the growth of the sugarcane in the example 1; similarly, in the comparative example 8, cercospora species is replaced by the same amount of actinomycetes, in the comparative example 9, alginic acid is replaced by the same amount of fulvic acid, and the performance detection data of the sugarcane obtained by using the composite microbial inoculum prepared in the comparative examples 8 and 9 is similar to that of the sugarcane using the comparative example 7, which indicates that the replacement of bacillus licheniformis, cercospora species and alginic acid can play a certain synergistic effect, and the replaced substances cannot play the synergistic effect. The inventor analyzes that: the bacillus licheniformis has stronger activities of protease, lipase and amylase and can promote the absorption and utilization of plants; cercospora micrantha produced by Cercospora has certain stimulation effect on the biological activity of plants, alginic acid can strengthen cell walls and has stable characteristic, and the cercospora micrantha produced by Cercospora and Cercospora micrantha are mixed and mutually promoted, so that the activities of enzyme and hormone required by the growth of the plants can be stimulated in a large range, and the plants can efficiently grow.
In the embodiments 1-6, the regulator is one or more of ethephon, fulvic acid, glyphosate and gibberellin, and the yield and sugar content of the sugarcane obtained by using the composite microbial inoculum prepared in the embodiments 1-6 are similar; in examples 7 and 8, the mass ratio is (0.5-0.8): (1-1.1): when fulvic acid, glyphosate and gibberellin in the range of (1.2-1.5) are mixed to be used as a regulator, the plant height, the stem diameter, the sugar content and the field brix of the obtained sugarcane are obviously improved, and the compound microbial inoculum prepared in examples 7 and 8 has stronger gain effect on the sugarcane yield and the sugar content.
The inventor analyzes that: the fulvic acid has the effects of promoting plant growth, particularly properly controlling the opening degree of leaf stomata of crops, reducing transpiration and improving quality, improves the activity of plant roots, improves various enzyme activities and increases the chlorophyll content by more than 20%. Glyphosate mainly inhibits enolpyruvylshikimate phosphate synthase in plants, thereby inhibiting the conversion of shikimate to phenylalanine, tyrosine and tryptophan and interfering the protein synthesis. Gibberellin can accelerate cell elongation and promote cell division. The three are mixed and cooperated in a certain proportion, so that the composite microbial inoculum A, the composite microbial inoculum B and the regulator can fully play a role, the decomposition and accumulation of the cane sugar are in a high-speed conversion balance state, and the yield and the sugar content of the cultivated cane sugar are improved.
In the embodiment 9, azotobacter, phosphate solubilizing bacteria and potassium solubilizing bacteria are also added into the composite microbial inoculum B; the compound microbial inoculum prepared in the embodiment 9 plays a role in increasing the growth and sugar content of sugarcane; the addition of the three beneficial bacteria further promotes the generation and conversion of elements such as nitrogen, phosphorus, potassium and the like in the soil, and further contributes to the promotion of the growth of sugarcane and the accumulation of sucrose. In examples 10 and 11, azospirillum and paspalum azotobacter which have directional effects on sugarcane are also selected respectively, so that the sugarcane obtained by using the composite microbial inoculum prepared in examples 10 and 11 has better growth and higher sugar content.
In example 12, fulvic acid, glyphosate and gibberellin which are mixed in a specific ratio are selected as regulators, and azotobacter, phosphate solubilizing bacteria and potassium solubilizing bacteria are also added into the composite microbial inoculum B, so that the composite microbial inoculum prepared in example 12 has the best effect on growth and sugar content gain of sugarcane, and example 12 is the best example of the application.
Test No. two
(1) Planting time: 2021, 4 months-12 months;
(2) planting place: the pond area of the pond is camphorwood for Deng Bao cun, Zhongtang village and Huanghe village;
(3) the test varieties are: sweet osmanthus No. 44;
(4) scheme design: 1. planting Saccharum sinensis Roxb No. 44 perennial root in Deng Bao village and Zhongtang village;
2. planting Arctose No. 44 in Huanghe village;
(5) sample preparation: control zone-commercial compound fertilizer 125 kg;
an experimental area, namely 125kg of a commercial compound fertilizer and 2kg of a microbial compound inoculant (the microbial compound inoculant prepared in the example 1 is selected);
(6) as a result: the test data are detailed in table 3;
TABLE 3
Experiment three
(1) Planting time: 2021, 4 months-12 months;
(2) planting place: a double-height demonstration base of a stone ballast town in the area of the pond in Guihong City;
(3) variety: cassia twig 05/136;
(4) scheme design: planting perennial roots;
(5) sample preparation: the contrast area-the planting base prepares fertilizer by itself;
2kg of fertilizer and microbial compound bacteria are prepared in an experimental area-a planting base (the microbial compound bacteria are prepared in the embodiment 1);
(6) as a result: the test detection data are detailed in tables 4 and 5;
TABLE 4 comparison of field test data
TABLE 5 comparison of sugar refinery laboratory test data
As can be seen from the data in tables 4 and 5, the sugarcane using the microbial composite inoculant prepared in example 1 has improved performance in all aspects, good growth, higher sugar content and greatly improved yield and sugar content of the sugarcane compared with the sugarcane without the microbial composite inoculant, and thus can bring considerable economic benefits.
The above embodiments are preferred embodiments of the present application, and the protection scope of the present application is not limited by the above embodiments, so: all equivalent changes made according to the structure, shape and principle of the present application shall be covered by the protection scope of the present application.
Claims (8)
1. A microbial compound inoculant for sugarcane planting is characterized in that: the composite material comprises the following components in percentage by mass:
35-40wt% of compound microbial inoculum A;
50-55wt% of compound microbial inoculum B;
5-15wt% of a regulator;
the complex microbial inoculum A comprises 55-60wt% of bacillus licheniformis suspension, 25-30wt% of cercospora suspension and 10-20wt% of alginic acid;
the preparation method of the complex microbial inoculum A comprises the following steps:
s1-1, respectively performing solid activation culture on the Bacillus licheniformis and Cercospora species to obtain the final product with concentration of 1-3 × 108CFU/ml of Bacillus licheniformis suspension liquid and Cercospora species suspension liquid;
s1-2, inoculating the bacillus licheniformis suspension liquid to a PDA culture medium by an inoculation amount of 5-7%, performing single-strain shake flask fermentation culture on a shaking table at the culture temperature of 32-35 ℃, the shaking speed of the shaking table of 150-170rpm, and the culture time of 72-76h to obtain the bacillus licheniformis suspension;
s1-3, inoculating the cercospora suspension liquid to a PDA culture medium in an inoculation amount of 3-5%, carrying out single-plant shake flask fermentation culture on a shaking table at the culture temperature of 32-35 ℃, the shaking table oscillation speed of 150-170rpm, and the culture time of 72-76h to obtain the cercospora suspension;
s1-4, mixing the bacillus licheniformis suspension, the cercospora suspension and alginic acid, adding into a stirring pot, and stirring at a constant speed of 100r/min for 15-20min to obtain a composite microbial inoculum A;
the complex microbial inoculum B at least comprises 45-50wt% of Bacillus mucilaginosus bacterial suspension, 25-30wt% of photosynthetic bacteria bacterial suspension and 16-30wt% of silicate bacteria bacterial suspension;
the preparation method of the composite microbial inoculum B comprises the following steps:
s2-1, respectively performing solid activation culture on Bacillus mucilaginosus strain, photosynthetic bacteria strain and silicate bacteria strain to obtain the culture solution with concentration of 1-3 × 108CFU/ml of Bacillus mucilaginosus suspension liquid, photosynthetic bacteria suspension liquid and silicate bacteria suspension liquid;
s2-2, inoculating the Bacillus mucilaginosus bacterial suspension liquid to a PDA culture medium in an inoculation amount of 5-7%, performing single-plant shake flask fermentation culture on a shaking table at the culture temperature of 32-35 ℃, the shaking table oscillation speed of 150-170rpm, and the culture time of 72-76h to obtain the Bacillus mucilaginosus bacterial suspension;
s2-3, inoculating the photosynthetic bacteria suspension liquid to an RCVBN culture medium by 3-5% of inoculation amount, carrying out single plant shake flask fermentation culture on a shaking table, wherein the culture temperature is 32-35 ℃, the shaking table oscillation speed is 150-170rpm, and the culture time is 72-76h, so as to obtain photosynthetic bacteria suspension;
s2-4, inoculating the silicate bacteria suspension liquid to an HB8548-2 culture medium in an inoculation amount of 2-4%, carrying out single-plant shake flask fermentation culture on a shaking table at the culture temperature of 30-32 ℃, the shaking table oscillation speed of 200-220rpm, and the culture time of 48-50h to obtain the silicate bacteria suspension;
s2-5, mixing the colloidal bacillus bacteria suspension, the photosynthetic bacteria suspension and the silicate bacteria suspension, adding into a stirring pot, and stirring at a constant speed of 100r/min for 15-20min to obtain the composite microbial agent B.
2. The microbial composite inoculant for sugarcane planting according to claim 1, wherein: the composite microbial inoculum A comprises the following components in percentage by mass: 57-58wt% of bacillus licheniformis suspension, 27-28wt% of cercospora suspension and 14-16wt% of alginic acid; the composite microbial inoculum B at least comprises the following components in percentage by mass: 47-50wt% of Bacillus mucilaginosus bacterial suspension, 25-28wt% of photosynthetic bacteria bacterial suspension and 16-22wt% of silicate bacteria bacterial suspension.
3. The microbial composite inoculant for sugarcane planting according to claim 2, wherein: the composite microbial inoculum A comprises the following components in percentage by mass: 57.5wt% of a Bacillus licheniformis suspension, 27.5wt% of a Cercospora suspension and 15wt% of alginic acid; the composite microbial inoculum B at least comprises the following components in percentage by mass: 48.5wt% Bacillus mucilaginosus bacterial suspension, 26.5wt% photosynthetic bacteria bacterial suspension and 19wt% silicate bacteria bacterial suspension.
4. The microbial composite inoculant for sugarcane planting according to any one of claims 1 to 3, wherein: the regulator is one or a mixture of fulvic acid, glyphosate and gibberellin.
5. The microbial composite inoculant for sugarcane planting according to claim 4, wherein: the regulator is prepared by mixing fulvic acid, glyphosate and gibberellin according to the mass ratio of (0.5-0.8) to (1-1.1) to (1.2-1.5).
6. The microbial composite inoculant for sugarcane planting according to any one of claims 1 to 3, wherein: the composite microbial inoculum B also comprises the following components in percentage by mass: 3-5wt% azotobacter suspension, 2-4wt% phosphate solubilizing bacteria suspension and 3-5wt% potassium solubilizing bacteria suspension;
the preparation method of the azotobacter bacterial suspension comprises the following steps: activating and culturing azotobacteria strain to obtain the product with concentration of 1-3 × 108CFU/ml azotobacter suspension liquid; inoculating the azotobacter suspension liquid to No. 77 azotobacter culture medium with the inoculation amount of 1-3%, performing single-strain shake flask fermentation culture on a shaking table at the culture temperature of 28-30 ℃, the shaking table oscillation speed of 200-220rpm, and the culture time of 48-50h to obtain azotobacter suspension;
the preparation method of the phosphate solubilizing bacterium suspension comprises the following steps: activating and culturing solid of phosphate solubilizing bacteria strain to obtain the product with concentration of 1-3 × 108CFU/ml of phosphate solubilizing bacteria suspension liquid; inoculating the phosphate solubilizing bacteria suspension liquid to an HB8549-2 phosphate solubilizing bacteria culture medium by an inoculation amount of 1-3%, performing single-strain shake flask fermentation culture on a shaking table at the culture temperature of 30-32 ℃, the shaking table oscillation speed of 200-220rpm, and the culture time of 48-50h to obtain the phosphate solubilizing bacteria suspension;
the preparation method of the potassium bacteria dissolving suspension comprises the following steps: activating and culturing solid potassium-decomposing bacteria strain to obtain the product with concentration of 1-3 × 108CFU/ml of potassium bacteria suspension liquid; inoculating the suspension liquid of the potassium bacteria to a potassium solution culture medium by the inoculation amount of 1-3%, and performing single-plant shake flask fermentation culture on a shaking table at the culture temperature of 37-40 ℃, the shaking table oscillation speed of 200-220rpm and the culture time of 48-50h to obtain the suspension of the potassium bacteria.
7. The microbial composite inoculant for sugarcane planting according to claim 6, wherein: the azotobacter strain is azospirillum strain or paspalum azotobacter strain.
8. A method for preparing a microbial composite inoculant for sugarcane planting according to any one of claims 1 to 7, wherein the microbial composite inoculant comprises the following components: the method comprises the following steps: and (3) uniformly mixing the compound microbial inoculum A, the compound microbial inoculum B and the regulator, and fermenting to obtain the microbial compound microbial inoculum.
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| WO2023151580A1 (en) * | 2022-02-10 | 2023-08-17 | 深圳市爱格丽生物科技有限公司 | Microbial complex microbial agent for sugarcane planting, and preparation method therefor |
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