WO2023151580A1 - Microbial complex microbial agent for sugarcane planting, and preparation method therefor - Google Patents

Microbial complex microbial agent for sugarcane planting, and preparation method therefor Download PDF

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WO2023151580A1
WO2023151580A1 PCT/CN2023/074969 CN2023074969W WO2023151580A1 WO 2023151580 A1 WO2023151580 A1 WO 2023151580A1 CN 2023074969 W CN2023074969 W CN 2023074969W WO 2023151580 A1 WO2023151580 A1 WO 2023151580A1
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suspension
bacterial agent
composite
microbial
agent
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PCT/CN2023/074969
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French (fr)
Chinese (zh)
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王剑英
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深圳市爱格丽生物科技有限公司
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G22/00—Cultivation of specific crops or plants not otherwise provided for
    • A01G22/55—Sugar cane
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00—Other organic fertilisers
    • C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor

Definitions

  • the present application relates to the field of microbial composite bacterial agents, in particular to a microbial composite bacterial agent for sugarcane planting and a preparation method thereof.
  • Sugarcane is a temperate and tropical crop, and sucrose is one of the final products of photosynthesis in sugarcane, and it is also the main form of transport and distribution of photosynthetic products in most plants.
  • "Double high" sugarcane refers to sugarcane with high yield and high sugar content.
  • sugarcane As a raw material for making sucrose and refining ethanol, sugarcane has high economic benefits and has been widely planted and promoted in southern my country.
  • Sugarcane is the raw material of the sugar industry, and the sugar content of sugarcane is the source of the economic benefits of the sugar industry.
  • the economic benefits of the sugar industry refer to the three main factors, the amount of cane crushed, the sugar production rate, and the cost per ton of sugar, which are interrelated and complement each other through financial accounting to comprehensively reflect the total profit of sugar products and the profit of 10,000 tons of sugarcane.
  • the amount of cane crushed is the foundation, which is related to the profit of 10,000 tons of cane.
  • the crushing period will be short, the sugar yield will be small, the utilization rate of equipment will be low, and the economic benefits will not be high.
  • the sugar production rate is low, the cost is high, and the economic benefit is not large. Only with sufficient and high-quality raw material cane can the greatest economic benefits be obtained.
  • the application provides a microbial Compound microbial agent and preparation method thereof.
  • the present application provides a microbial composite bacterial agent for planting sugarcane, which adopts the following technical scheme:
  • a microbial compound bacterial agent for planting sugarcane comprising the following components in mass percentage: Composite microbial agent A 35-40wt%; Composite bacterial agent B 50-55wt%; Regulator 5-15wt%;
  • the composite bacterial agent A includes 55-60wt% of Bacillus licheniformis suspension, 25-30wt% of Cercospora suspension and 10-20wt% of alginic acid;
  • the preparation method of described composite bacterial agent A comprises:
  • Bacillus licheniformis suspension seed liquid and Cercospora bacterial suspension seed liquid were prepared with a concentration of 1-3 ⁇ 10 8 CFU/ml liquid;
  • the composite bacterial agent B includes 25-30wt% of the photosynthetic bacteria suspension and the jelly-like Paenibacillus suspension balance; wherein, the photosynthetic bacteria can be Rhodopseudomonas palustris, Rhodopseudomonas capsule middle a kind of
  • the preparation method of described composite bacterial agent B comprises:
  • the composite microbial agent A comprises the following components by mass percentage: 57-58wt% of Bacillus licheniformis suspension, 27-28wt% of Cercospora suspension and 14-16wt% of alginic acid;
  • the composite microbial agent B includes the following components in mass percentage: 25-28 wt% of photosynthetic bacteria suspension and the balance of jelly-like Paenibacillus suspension.
  • the composite bacterial agent A comprises the following components by mass percentage: 57.5wt% of the Bacillus licheniformis bacterial suspension, 27.5wt% of the Cercospora bacterial suspension and 15wt% alginic acid; B at least includes the following components in mass percentage: 26.5wt% photosynthetic bacteria suspension and the balance of jelly-like Paenibacillus suspension.
  • Sugarcane sugar accumulation is a complex process regulated by a variety of sucrose metabolizing enzymes, involving sucrose Physiological processes such as synthesis, breakdown and transport of sugars. Enzymes can catalyze the synthesis of sucrose and regulate sucrose content. Among them, sucrose phosphate synthase (SPS), sucrose synthase (SS) and invertase (INV) are the three key enzymes regulating sucrose metabolism. SPS is responsible for the synthesis of sucrose; SS is a reversible enzyme that can not only catalyze the synthesis of sucrose, but also catalyze the decomposition of sucrose; INV plays an important role in the transport, storage and distribution of sucrose. In plants, SPS expressed in both source and sink is involved in sucrose resynthesis, regulation of starch accumulation, protein storage and cellulose synthesis.
  • SPS sucrose phosphate synthase
  • SS sucrose synthase
  • INV invertase
  • the joint action of Bacillus licheniformis, Cercospora and alginic acid in the composite bacterial agent A can enhance the activity of amylase, protease and invertase (INV), and promote the decomposition and transformation of sucrose and nutrients, It makes nutrients more easily absorbed by plants and helps to promote the growth of sugarcane plants; the jelly-like Paenibacillus and photosynthetic bacteria in the compound microbial agent B work together to increase the nitrogen, potassium, phosphorus, boron and other nutrients in the soil.
  • the formation and transformation of elements have a positive effect on the accumulation of sucrose in sugarcane; the addition of regulators can stimulate the metabolic enzymes in sugarcane to a certain extent, and the regulator, composite bacterial agent A and composite bacterial agent B are in a certain proportion Mixing and working together help to better balance the accumulation or transformation of sucrose in sugarcane, enabling the dynamic balance of high-speed transformation of sucrose, and further promoting the growth of sugarcane and the accumulation of sucrose, thereby realizing the "high yield, high yield" of sugarcane sugar” target.
  • the joint action of the various components within the three is also conducive to improving the inhibitory resistance of sugarcane to external fungi and bacteria.
  • the regulator is one or more of ethephon, chitosan and polyglutamic acid.
  • the preparation method of a kind of microbial composite bacterial agent for sugarcane planting provided by the application adopts the following technical scheme:
  • a method for preparing a microbial composite bacterial agent for planting sugarcane comprising the following steps: uniformly mixing composite microbial agent A, composite bacterial agent B and a regulator to obtain a microbial composite bacterial agent.
  • the composite bacterial agent A and the composite bacterial agent B are respectively prepared earlier, so that the internal components of the composite bacterial agent A and the composite bacterial agent B can be uniformly mixed, which is beneficial to the composite bacterial agent A, the composite bacterial agent
  • the components inside the bacterial agent B work together to improve the respective effects of the composite bacterial agent A and the composite bacterial agent B; finally, the regulator is evenly mixed with the composite bacterial agent A and the composite bacterial agent B, so that the microbial composite bacterial agent system reaches The balance is stable, so that the sugarcane yield and sugar content of the application of the microbial compound bacterial agent are both improved.
  • Bacillus licheniformis, Cercospora and alginic acid contained in the composite bacterial agent A in the present application work together to help promote the plant growth of sugarcane;
  • the jelly-like Paenibacillus and the Photosynthetic bacteria work together, which is beneficial to the formation and stability of nitrogen, potassium, phosphorus, boron and other elements in the soil, and can promote the accumulation of sucrose in sugarcane;
  • the regulator is mainly plant growth hormone;
  • the regulator, compound bacterial agent A Mixed with the compound bacterial agent B they work together to help better control the accumulation or transformation of sucrose in sugarcane, and then achieve the goal of "high yield and high sugar content" of sugarcane;
  • ethephon, oligochitosan and polyglutamic acid are selected as regulators.
  • Ethephon, oligochitosan and polyglutamic acid can be more effective than other regulators when used in conjunction with compound bacterial agents A and B.
  • Bacillus licheniformis, Cercospora, Paenibacillus jelly-like, photosynthetic bacteria species required in the Examples and Comparative Examples of the present application and the corresponding culture medium were obtained commercially.
  • This embodiment discloses a kind of microbial composite bacteria agent for sugarcane plantation, including the following raw materials:
  • Composite bacterial agent A comprises 55wt% of bacillus licheniformis bacterial suspension, 25wt% of cercospora bacterial suspension and 20wt% of alginic acid;
  • the preparation method of composite microbial agent A is:
  • Bacillus licheniformis suspension seed liquid and Cercospora bacterial suspension seed liquid were prepared with a concentration of 1-3 ⁇ 10 8 CFU/ml liquid;
  • Composite microbial agent B includes 75 wt% of jelly-like Paenibacillus suspension and 25 wt% of photosynthetic bacteria suspension.
  • the preparation method of composite microbial agent B is:
  • the present embodiment also discloses a preparation method of a microbial composite bacterial agent for sugarcane plantation, comprising the following steps:
  • This embodiment discloses a kind of microbial composite bacteria agent for sugarcane plantation, including the following raw materials:
  • the preparation method of composite microbial agent A is:
  • Bacillus licheniformis suspension seed liquid and Cercospora bacterial suspension seed liquid were prepared with a concentration of 1-3 ⁇ 10 8 CFU/ml liquid;
  • the preparation method of composite microbial agent B is:
  • the present embodiment also discloses a preparation method of a microbial composite bacterial agent for sugarcane plantation, comprising the following steps:
  • This embodiment discloses a kind of microbial composite bacteria agent for sugarcane plantation, including the following raw materials:
  • Composite bacterial agent A comprises 60wt% of bacillus licheniformis suspension, 30wt% of cercospora bacterial suspension and 10wt% of alginic acid; the preparation method of composite bacterial agent A is:
  • Bacillus licheniformis suspension seed liquid and Cercospora bacterial suspension seed liquid were prepared with a concentration of 1-3 ⁇ 10 8 CFU/ml liquid;
  • Composite microbial agent B includes 70 wt% of jelly-like Paenibacillus suspension and 30 wt% of photosynthetic bacteria suspension.
  • the preparation method of composite microbial agent B is:
  • the present embodiment also discloses a preparation method of a microbial composite bacterial agent for sugarcane plantation, comprising the following steps:
  • a microbial compound bacterial agent for sugarcane planting differs from Example 1 in that it does not contain compound bacterial agent B and regulator.
  • a kind of microbial compound bacterial agent that is used for sugarcane plantation is: not containing compound bacterial agent A and regulator.
  • a microbial composite bacterial agent for sugarcane planting differs from Example 1 in that it does not contain composite microbial agent A and composite bacterial agent B.
  • a microbial compound bacterial agent for sugarcane planting differs from Example 1 in that it does not contain regulators.
  • a microbial composite bacterial agent for sugarcane planting the difference from Example 1 is: no composite bacterial agent B.
  • a microbial compound bacterial agent used for sugarcane planting differs from Example 1 in that it does not contain compound bacterial agent A.
  • a kind of microbial composite bacterial agent that is used for sugarcane planting, and the difference of embodiment 1 is: the suspension of Bacillus licheniformis is replaced by the suspension of Bacillus amyloliquefaciens of equal amount.
  • a kind of microbial compound bacterial agent that is used for sugarcane plantation the difference with embodiment 1 is: the cercospora suspension is replaced by the actinomycetes suspension of equal amount, the preparation method of actinomyces suspension The preparation method is the same as that of the Cercospora suspension, and the medium is PDA medium.
  • a microbial compound bacterial agent for sugarcane cultivation the difference from Example 1 is that alginic acid is replaced by an equivalent amount of fulvic acid.
  • the experimental field selected by Huanghe Village is divided into 13 equal parts, which are recorded as 1-1, 1-2, 1-3, etc.; 10 new plants of Guitang No. 44 are planted in each part;
  • the microbial compound microbial agent that makes among the embodiment 1-3 comprises three major components of composite microbial agent A, composite bacterial agent B and regulator, and the mutual promotion between the three major components works together, so that using embodiment 1 Compared with the sugarcane of the control group, the sugarcane of the compound bacterial agent prepared in -3 has better yield and higher sugar content, and is closer to the planting goal of "double high" sugarcane.
  • Composite bacterial agent B and regulator were not contained in comparative example 1, compared with blank control group, comparative example 1 added composite bacterial agent A, and the sugarcane obtained by using the composite bacterial agent obtained in comparative example 1 was compared with the sugarcane obtained by the control
  • the plant height, stem diameter, sucrose content and field hammer have all been improved, especially the improvement of plant height and stem diameter is more obvious, indicating that the compound bacterial agent A has the effect of promoting the growth of sugarcane;
  • comparative example 2 does not contain the compound bacterial agent A and regulator, compared with the control group in comparative example 2, the compound microbial agent B was added more.
  • the sugarcane obtained by using the compound bacterial agent obtained in comparative example 2 had higher plant height, stem diameter, sucrose The content and field Brix have also been improved, especially the increase of sucrose content and field Brix is more obvious, indicating that the compound bacterial agent B has the effect of increasing the sucrose content of sugarcane.
  • Comparing example 4 does not contain regulator, comparative example 4 has added composite bacterial agent A and composite bacterial agent B more compared to the control group, and the sugarcane obtained by using comparative example 4 to make composite bacterial agent is compared with using comparative examples 1 and 2 Obtained sugarcane plant height, stem diameter, sucrose content and field Brix have a small increase, indicating that composite bacterial agent A and composite bacterial agent B may produce mutual inhibition when they are used together;
  • comparative example 5 is a composite bacterial agent A is mixed with regulator, and comparative example 6 is that compound bacterial agent B is mixed with regulator, and the performance test data values of comparative examples 5 and 6 are also higher than the performance test data values of comparative example 4, further reflecting compound bacterial agent A and compound bacterial agent Inhibitory effect between the two agents B.
  • Bacillus licheniformis has strong protease, lipase, and amylase activities, which can promote the absorption and utilization of plants; the cercosporin produced by Cercospora has a certain stimulating effect on the biological activity of plants, and alginic acid can It strengthens the cell wall and has stable characteristics. After the three are mixed, they promote each other and can stimulate the activity of enzymes and hormones required for plant growth in a large range, so that plants can grow efficiently.
  • Example 1-3 ethephon was selected as the regulator, and the yield and sugar content of sugarcane obtained by using the compound bacterial agent prepared in Example 1-3 were similar, and had a good effect on the yield and sugar content of sugarcane.
  • Composite bacterial agent A, composite bacterial agent B and ethephon can fully play a role, so that the decomposition and accumulation of sucrose are in a high-speed transformation equilibrium state, and then the yield and sugar content of the cultivated sugarcane are improved.
  • control area commercially available compound fertilizer 125kg
  • control area the planting base mixes its own fertilizer
  • Experimental area-planting base oneself mixes fertilizer+microbial composite bacterial agent 2kg (microbial composite bacterial agent selects the microbial composite bacterial agent that embodiment 1 makes for use);
  • the sugarcane that used the microbial composite bacterial agent obtained in Example 1 has improved in all aspects, with good growth and sugar content.
  • the higher quality greatly increases the yield and sugar content of sugarcane, which in turn can bring considerable economic benefits.

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Abstract

Provided are a microbial complex microbial agent for sugarcane planting, and a preparation method therefor. The microbial complex microbial agent comprises the following components in percentage by mass: 35-40wt% of a complex microbial agent A, 50-55wt% of a complex microbial agent B, and 5-15wt% of an adjustment agent; the complex microbial agent A comprises 55-60wt% of a bacillus licheniformis suspension, 25-30wt% of a cercospora suspension, and 10-20wt% of alginic acid; and the complex microbial agent B comprises 25-30wt% of a photosynthetic bacteria suspension and the balance being a paenibacillus mucilaginosus suspension. The microbial agent has the effects of improving the yield and sugar content of sugarcane.

Description

一种用于甘蔗种植的微生物复合菌剂及其制备方法A kind of microbial composite bacteria agent for sugarcane planting and preparation method thereof 技术领域technical field
本申请涉及微生物复合菌剂领域，尤其是涉及一种用于甘蔗种植的微生物复合菌剂及其制备方法。The present application relates to the field of microbial composite bacterial agents, in particular to a microbial composite bacterial agent for sugarcane planting and a preparation method thereof.
背景技术Background technique
甘蔗是温带和热带农作物，蔗糖是甘蔗光合作用的最终产物之一，也是绝大多数植物体内光合产物运输与分配的主要形式。“双高”甘蔗是指产量高、含糖量高的甘蔗。甘蔗作为制作蔗糖和提炼乙醇的原料，具有较高的经济效益，在我国南方地区得到了广泛的种植与推广。Sugarcane is a temperate and tropical crop, and sucrose is one of the final products of photosynthesis in sugarcane, and it is also the main form of transport and distribution of photosynthetic products in most plants. "Double high" sugarcane refers to sugarcane with high yield and high sugar content. As a raw material for making sucrose and refining ethanol, sugarcane has high economic benefits and has been widely planted and promoted in southern my country.
甘蔗是制糖工业的原料，甘蔗蔗糖分是制糖工业经济效益的源泉。制糖工业经济效益就是指榨蔗量、产糖率和吨糖成本三个主要因素互相联系，相辅相成地通过财务核算,综合反映出来的糖品总利润和万吨蔗利润。榨蔗量是基础，关系到万吨蔗利润的高低,但是只有蔗糖含量高而没有一定数量的原料蔗,则榨期短,产糖量少,设备利用率低，经济效益不高。而虽有一定数量的原料蔗但它的蔗糖分低，则产糖率低，成本高，经济效益也不大。只有数量充足且优质的原料蔗，才能获得最大的经济效益。Sugarcane is the raw material of the sugar industry, and the sugar content of sugarcane is the source of the economic benefits of the sugar industry. The economic benefits of the sugar industry refer to the three main factors, the amount of cane crushed, the sugar production rate, and the cost per ton of sugar, which are interrelated and complement each other through financial accounting to comprehensively reflect the total profit of sugar products and the profit of 10,000 tons of sugarcane. The amount of cane crushed is the foundation, which is related to the profit of 10,000 tons of cane. However, if there is only a high content of sucrose without a certain amount of raw cane, the crushing period will be short, the sugar yield will be small, the utilization rate of equipment will be low, and the economic benefits will not be high. And although there is a certain amount of raw material cane but its sucrose content is low, then the sugar production rate is low, the cost is high, and the economic benefit is not large. Only with sufficient and high-quality raw material cane can the greatest economic benefits be obtained.
现有的农业种植中，无毒无害、无环境污染的微生物肥料的广泛使用，针对上述技术问题，发明人认为通过微生物肥料中微生物生命代谢活动调节和改善甘蔗的产量及含糖量，得到产量高、含糖量高的“双高”甘蔗，是未来甘蔗种植中重点研究的方向。In the existing agricultural planting, non-toxic, harmless, and non-environmentally polluting microbial fertilizers are widely used. In view of the above-mentioned technical problems, the inventor believes that the yield and sugar content of sugarcane can be adjusted and improved through microbial life and metabolic activities in microbial fertilizers to obtain "Double-high" sugarcane with high yield and high sugar content is the key research direction in future sugarcane planting.
发明内容Contents of the invention
为了提升甘蔗的产量和含糖量，本申请提供一种用于甘蔗种植的微生 物复合菌剂及其制备方法。In order to increase the yield and sugar content of sugarcane, the application provides a microbial Compound microbial agent and preparation method thereof.
第一方面，本申请提供的一种用于甘蔗种植的微生物复合菌剂,采用如下的技术方案：In the first aspect, the present application provides a microbial composite bacterial agent for planting sugarcane, which adopts the following technical scheme:
一种用于甘蔗种植的微生物复合菌剂，包括以下质量百分比的组分：
复合菌剂A 35-40wt％；
复合菌剂B 50-55wt％；
调节剂5-15wt％；A microbial compound bacterial agent for planting sugarcane, comprising the following components in mass percentage:
Composite microbial agent A 35-40wt%;
Composite bacterial agent B 50-55wt%;
Regulator 5-15wt%;
所述复合菌剂A包括55-60wt％的地衣芽孢杆菌菌悬液、25-30wt％的尾孢菌菌悬液以及10-20wt％的海藻酸；The composite bacterial agent A includes 55-60wt% of Bacillus licheniformis suspension, 25-30wt% of Cercospora suspension and 10-20wt% of alginic acid;
所述复合菌剂A的制备方法包括：The preparation method of described composite bacterial agent A comprises:
S1-1，将地衣芽孢杆菌和尾孢菌菌株分别固体活化培养后，制得浓度均为1-3×108CFU/ml的地衣芽孢杆菌菌悬液种液和尾孢菌菌悬液种液；S1-1, after solid activation and culture of Bacillus licheniformis and Cercospora strains respectively, Bacillus licheniformis suspension seed liquid and Cercospora bacterial suspension seed liquid were prepared with a concentration of 1-3×10 8 CFU/ml liquid;
S1-2，将地衣芽孢杆菌菌悬液种液以5-7％的接种量接种至液体培养基，培养温度为32-35℃，培养时间为72-76h，140-200rpm振荡培养得到地衣芽孢杆菌菌悬液；S1-2, inoculate the Bacillus licheniformis suspension seed liquid into the liquid medium with an inoculum amount of 5-7%, the culture temperature is 32-35°C, the culture time is 72-76h, and the 140-200rpm shaking culture is obtained to obtain the licheniformis spores Bacillus suspension;
S1-3，将尾孢菌菌悬液种液以3-5％的接种量接种至PDA培养基，培养温度为32-35℃，培养时间为72-76h，140-200rpm振荡培养得到尾孢菌菌悬液；S1-3, inoculate the seed liquid of the Cercospora suspension to the PDA medium with an inoculum amount of 3-5%, the culture temperature is 32-35°C, the culture time is 72-76h, and the cercospora is obtained by shaking culture at 140-200rpm bacterial suspension;
S1-4，将地衣芽孢杆菌菌悬液、尾孢菌菌悬液和海藻酸混合，得到复合菌剂A；S1-4, mixing Bacillus licheniformis suspension, Cercospora suspension and alginic acid to obtain composite microbial agent A;
所述复合菌剂B包括25-30wt％的光合细菌菌悬液以及胶冻样类芽孢杆菌菌悬液余量；其中，光合细菌可为沼泽红假单胞菌、荚膜红假单胞菌中 的一种；The composite bacterial agent B includes 25-30wt% of the photosynthetic bacteria suspension and the jelly-like Paenibacillus suspension balance; wherein, the photosynthetic bacteria can be Rhodopseudomonas palustris, Rhodopseudomonas capsule middle a kind of
所述复合菌剂B的制备方法包括：The preparation method of described composite bacterial agent B comprises:
S2-1，将胶冻样类芽孢杆菌菌种和光合细菌菌种分别固体活化培养后，制得浓度均为1-3×108CFU/ml的胶冻样类芽孢杆菌菌悬液种液和光合细菌菌悬液种液；S2-1, after the jelly-like Paenibacillus strain and the photosynthetic bacteria strain are respectively activated and cultured on a solid basis, a jelly-like Paenibacillus suspension seed solution with a concentration of 1-3×10 8 CFU/ml is prepared and photosynthetic bacteria suspension seed liquid;
S2-2，将胶冻样类芽孢杆菌菌悬液种液以5-7％的接种量接种至液体培养基，培养温度为32-35℃，培养时间为72-76h，140-200rpm振荡培养得到胶冻样类芽孢杆菌菌悬液；S2-2, inoculate the jelly-like Paenibacillus suspension seed liquid into the liquid medium with an inoculum size of 5-7%, the culture temperature is 32-35°C, the culture time is 72-76h, and the culture is shaken at 140-200rpm Obtain jelly-like Paenibacillus suspension;
S2-3，将光合细菌菌悬液种液以3-5％的接种量接种至RCVBN培养基，培养温度为32-35℃，培养时间为72-76h，140-200rpm振荡培养得到光合细菌菌悬液；S2-3, inoculate the seed solution of the photosynthetic bacteria suspension into the RCVBN medium with an inoculum size of 3-5%, the culture temperature is 32-35°C, the culture time is 72-76h, and the photosynthetic bacteria are obtained by shaking culture at 140-200rpm suspension;
S2-4，将胶冻样类芽孢杆菌菌悬液和光合细菌菌悬液混合,得到复合菌剂B。S2-4, mixing the jelly-like Paenibacillus suspension and the photosynthetic bacteria suspension to obtain a composite bacterial agent B.
优选的，所述复合菌剂A包括以下质量百分比的组分：57-58wt％的地衣芽孢杆菌菌悬液、27-28wt％的尾孢菌菌悬液以及14-16wt％的海藻酸；所述复合菌剂B包括以下质量百分比的组分：25-28wt％的光合细菌菌悬液以及胶冻样类芽孢杆菌菌悬液余量。Preferably, the composite microbial agent A comprises the following components by mass percentage: 57-58wt% of Bacillus licheniformis suspension, 27-28wt% of Cercospora suspension and 14-16wt% of alginic acid; The composite microbial agent B includes the following components in mass percentage: 25-28 wt% of photosynthetic bacteria suspension and the balance of jelly-like Paenibacillus suspension.
优选的，所述复合菌剂A包括以下质量百分比的组分：57.5wt％的地衣芽孢杆菌菌悬液、27.5wt％的尾孢菌菌悬液以及15wt％的海藻酸；所述复合菌剂B至少包括以下质量百分比的组分：26.5wt％的光合细菌菌悬液以及胶冻样类芽孢杆菌菌悬液余量。Preferably, the composite bacterial agent A comprises the following components by mass percentage: 57.5wt% of the Bacillus licheniformis bacterial suspension, 27.5wt% of the Cercospora bacterial suspension and 15wt% alginic acid; B at least includes the following components in mass percentage: 26.5wt% photosynthetic bacteria suspension and the balance of jelly-like Paenibacillus suspension.
甘蔗糖分积累是一个由多种蔗糖代谢酶参与调控的复杂过程，涉及蔗 糖的合成、分解和运输等生理过程。酶可以催化蔗糖合成的和调节蔗糖含量。其中蔗糖磷酸合成酶(SPS)、蔗糖合成酶(SS)和转化酶(INV)是调控蔗糖代谢的3大关键酶。SPS负责蔗糖的合成；SS是一种可逆酶，既能催化蔗糖合成，又能催化蔗糖分解；INV在蔗糖的转运、贮藏和分配中具有重要作用。在植物中，同时在源和库中表达的SPS参与蔗糖的再合成，调节淀粉积累、蛋白质储存和纤维素合成。Sugarcane sugar accumulation is a complex process regulated by a variety of sucrose metabolizing enzymes, involving sucrose Physiological processes such as synthesis, breakdown and transport of sugars. Enzymes can catalyze the synthesis of sucrose and regulate sucrose content. Among them, sucrose phosphate synthase (SPS), sucrose synthase (SS) and invertase (INV) are the three key enzymes regulating sucrose metabolism. SPS is responsible for the synthesis of sucrose; SS is a reversible enzyme that can not only catalyze the synthesis of sucrose, but also catalyze the decomposition of sucrose; INV plays an important role in the transport, storage and distribution of sucrose. In plants, SPS expressed in both source and sink is involved in sucrose resynthesis, regulation of starch accumulation, protein storage and cellulose synthesis.
通过采用上述技术方案，复合菌剂A中地衣芽孢杆菌、尾孢菌和海藻酸三者共同作用，能够增强淀粉酶、蛋白酶、转化酶(INV)的活性，促进蔗糖和营养物质的分解转化，使得营养物质更容易被植物吸收，有助于促进甘蔗的植株生长；复合菌剂B中胶冻样类芽孢杆菌和光合细菌共同作用，增加了土壤中的氮、钾、磷、硼等多种元素的生成和转化，对甘蔗中蔗糖含量的积累有增益效果；调节剂的加入能够一定程度上刺激甘蔗中的代谢酶，调节剂、复合菌剂A和复合菌剂B三者按照一定的比例混合，共同作用，有助于更好地平衡甘蔗中蔗糖的积累或转化，使得蔗糖实现高速转化的动态平衡，还进一步促进了甘蔗的生长与蔗糖的积累，进而实现甘蔗的“高产量、高糖分”的目标。三者内部各组分之间共同作用，还有利于提升甘蔗面对外界真菌、细菌的抑制抵抗效果。By adopting the above technical scheme, the joint action of Bacillus licheniformis, Cercospora and alginic acid in the composite bacterial agent A can enhance the activity of amylase, protease and invertase (INV), and promote the decomposition and transformation of sucrose and nutrients, It makes nutrients more easily absorbed by plants and helps to promote the growth of sugarcane plants; the jelly-like Paenibacillus and photosynthetic bacteria in the compound microbial agent B work together to increase the nitrogen, potassium, phosphorus, boron and other nutrients in the soil. The formation and transformation of elements have a positive effect on the accumulation of sucrose in sugarcane; the addition of regulators can stimulate the metabolic enzymes in sugarcane to a certain extent, and the regulator, composite bacterial agent A and composite bacterial agent B are in a certain proportion Mixing and working together help to better balance the accumulation or transformation of sucrose in sugarcane, enabling the dynamic balance of high-speed transformation of sucrose, and further promoting the growth of sugarcane and the accumulation of sucrose, thereby realizing the "high yield, high yield" of sugarcane sugar" target. The joint action of the various components within the three is also conducive to improving the inhibitory resistance of sugarcane to external fungi and bacteria.
优选的，所述调节剂为乙烯利、壳寡糖和聚谷氨酸中的一种或多种。Preferably, the regulator is one or more of ethephon, chitosan and polyglutamic acid.
通过采用上述技术方案，乙烯利、壳寡糖和聚谷氨酸能够良好的配合复合菌剂A、B使用，进而有效调控复合菌剂A和复合菌剂B对于蔗糖合成和分解的平衡性，进而缓解因生长过快导致蔗糖积累不充分或蔗糖转换不充分阻碍生长之间的矛盾，使得甘蔗生长良好，蔗糖含量也提升。 By adopting the above-mentioned technical scheme, ethephon, chitosan and polyglutamic acid can be well used in conjunction with composite bacterial agents A and B, and then effectively regulate the balance of composite bacterial agent A and composite bacterial agent B for the synthesis and decomposition of sucrose, Furthermore, the contradiction between insufficient sucrose accumulation or insufficient sucrose conversion hindering growth due to excessive growth is alleviated, so that the sugarcane grows well and the sucrose content is also increased.
第二方面，本申请提供的一种用于甘蔗种植的微生物复合菌剂的制备方法采用如下的技术方案：In the second aspect, the preparation method of a kind of microbial composite bacterial agent for sugarcane planting provided by the application adopts the following technical scheme:
一种用于甘蔗种植的微生物复合菌剂的制备方法，包括以下步骤：将复合菌剂A、复合菌剂B和调节剂混合均匀，得到微生物复合菌剂。A method for preparing a microbial composite bacterial agent for planting sugarcane, comprising the following steps: uniformly mixing composite microbial agent A, composite bacterial agent B and a regulator to obtain a microbial composite bacterial agent.
通过采用上述技术方案，先各自制得复合菌剂A、复合菌剂B，使得复合菌剂A、复合菌剂B中内部各组分之间可以均匀混合，进而有利于复合菌剂A、复合菌剂B内部各组分之间共同作用，提升复合菌剂A、复合菌剂B各自的效果；最后将调节剂和复合菌剂A、复合菌剂B混合均匀，使得微生物复合菌剂体系达到平衡稳定，进而使得施用微生物复合菌剂的甘蔗产量和糖分均得到提升。By adopting the above-mentioned technical scheme, the composite bacterial agent A and the composite bacterial agent B are respectively prepared earlier, so that the internal components of the composite bacterial agent A and the composite bacterial agent B can be uniformly mixed, which is beneficial to the composite bacterial agent A, the composite bacterial agent The components inside the bacterial agent B work together to improve the respective effects of the composite bacterial agent A and the composite bacterial agent B; finally, the regulator is evenly mixed with the composite bacterial agent A and the composite bacterial agent B, so that the microbial composite bacterial agent system reaches The balance is stable, so that the sugarcane yield and sugar content of the application of the microbial compound bacterial agent are both improved.
综上所述，本申请包括以下有益技术效果：In summary, the application includes the following beneficial technical effects:
1.本申请中复合菌剂A中含有的地衣芽孢杆菌、尾孢菌和海藻酸三者共同作用，有助于促进甘蔗的植株生长；复合菌剂B中含有的胶冻样类芽孢杆菌和光合细菌共同作用，有利于土壤中的氮、钾、磷、硼等多种元素的生成和稳定，能够促进甘蔗中蔗糖含量的积累；调节剂主要为植物生长激素；调节剂、复合菌剂A和复合菌剂B三者混合，共同作用，有助于更好地调配控制甘蔗中蔗糖的积累或转化，进而实现甘蔗的“高产量、高糖分”的目标；1. Bacillus licheniformis, Cercospora and alginic acid contained in the composite bacterial agent A in the present application work together to help promote the plant growth of sugarcane; the jelly-like Paenibacillus and the Photosynthetic bacteria work together, which is beneficial to the formation and stability of nitrogen, potassium, phosphorus, boron and other elements in the soil, and can promote the accumulation of sucrose in sugarcane; the regulator is mainly plant growth hormone; the regulator, compound bacterial agent A Mixed with the compound bacterial agent B, they work together to help better control the accumulation or transformation of sucrose in sugarcane, and then achieve the goal of "high yield and high sugar content" of sugarcane;
2.本申请中调节剂选用乙烯利、壳寡糖和聚谷氨酸，乙烯利、壳寡糖和聚谷氨酸配合复合菌剂A、B使用相较于其他的调节剂能够更加有效的调控复合菌剂A和复合菌剂B对于蔗糖合成和分解的平衡性，进而缓解因生长过快导致蔗糖积累不充分或蔗糖转换不充分阻碍生长之间的矛盾，使 得甘蔗生长良好，蔗糖含量也提升。2. In this application, ethephon, oligochitosan and polyglutamic acid are selected as regulators. Ethephon, oligochitosan and polyglutamic acid can be more effective than other regulators when used in conjunction with compound bacterial agents A and B. Regulate the balance between composite bacterial agent A and composite bacterial agent B on the synthesis and decomposition of sucrose, thereby alleviating the contradiction between insufficient sucrose accumulation or insufficient sucrose conversion hindering growth due to excessive growth, so that The sugar cane grows well, and the sucrose content also increases.
具体实施方式Detailed ways
以下结合实施例对本申请作进一步详细说明。The present application will be described in further detail below in conjunction with the examples.
市购获得本申请实施例、对比例中所需的地衣芽孢杆菌、尾孢菌、胶冻样类芽孢杆菌、光合细菌菌种以及对应需要的培养基。The Bacillus licheniformis, Cercospora, Paenibacillus jelly-like, photosynthetic bacteria species required in the Examples and Comparative Examples of the present application and the corresponding culture medium were obtained commercially.
实施例Example
实施例1Example 1
本实施例公开一种用于甘蔗种植的微生物复合菌剂，包括以下原料：This embodiment discloses a kind of microbial composite bacteria agent for sugarcane plantation, including the following raw materials:
3.5kg复合菌剂A；5.0kg复合菌剂B；1.5kg调节剂；调节剂为乙烯利。3.5kg composite bacterial agent A; 5.0kg composite bacterial agent B; 1.5kg regulator; regulator is ethephon.
复合菌剂A包括地衣芽孢杆菌菌悬液55wt％、尾孢菌菌悬液25wt％以及海藻酸20wt％；复合菌剂A的制备方法为：Composite bacterial agent A comprises 55wt% of bacillus licheniformis bacterial suspension, 25wt% of cercospora bacterial suspension and 20wt% of alginic acid; The preparation method of composite microbial agent A is:
S1-1，将地衣芽孢杆菌和尾孢菌菌株分别固体活化培养后，制得浓度均为1-3×108CFU/ml的地衣芽孢杆菌菌悬液种液和尾孢菌菌悬液种液；S1-1, after solid activation and culture of Bacillus licheniformis and Cercospora strains respectively, Bacillus licheniformis suspension seed liquid and Cercospora bacterial suspension seed liquid were prepared with a concentration of 1-3×10 8 CFU/ml liquid;
S1-2，将地衣芽孢杆菌菌悬液种液以5％的接种量接种至PDA液体培养基，在摇床上进行单株摇瓶发酵培养，培养温度为32℃，摇床振荡速率为150rpm，培养时间为72h，得到地衣芽孢杆菌菌悬液；S1-2, inoculate the Bacillus licheniformis suspension seed solution to the PDA liquid medium with an inoculum amount of 5%, carry out a single-plant shake flask fermentation culture on a shaker, the culture temperature is 32 ° C, and the shaker oscillation speed is 150 rpm, The culture time is 72 hours, and the suspension of Bacillus licheniformis is obtained;
S1-3，将尾孢菌菌悬液种液以3％的接种量接种至PDA培养基，在摇床上进行单株摇瓶发酵培养，培养温度为32℃，摇床振荡速率为150rpm，培养时间为72h，得到尾孢菌菌悬液；S1-3, inoculate the Cercospora suspension seed solution to the PDA medium with a 3% inoculum amount, carry out single-plant shake flask fermentation culture on a shaker, the culture temperature is 32°C, and the shaker oscillation speed is 150rpm. The time is 72h to obtain the suspension of Cercospora bacteria;
S1-4，将5.5kg地衣芽孢杆菌菌悬液、2.5kg尾孢菌菌悬液和2kg海藻酸混合加入搅拌锅中，以100r/min的速度匀速搅拌15min，得到复合菌 剂A；S1-4, mix 5.5kg of Bacillus licheniformis suspension, 2.5kg of Cercospora suspension and 2kg of alginic acid into the stirring pot, and stir at a constant speed of 100r/min for 15min to obtain composite bacteria Agent A;
复合菌剂B包括胶冻样类芽孢杆菌菌悬液75wt％和光合细菌菌悬液25wt％。复合菌剂B的制备方法为：Composite microbial agent B includes 75 wt% of jelly-like Paenibacillus suspension and 25 wt% of photosynthetic bacteria suspension. The preparation method of composite microbial agent B is:
S2-1，将胶冻样类芽孢杆菌菌种和光合细菌菌种分别固体活化培养后，制得浓度均为1-3×108CFU/ml的胶冻样类芽孢杆菌菌悬液种液和光合细菌菌悬液种液；S2-1, after the jelly-like Paenibacillus strain and the photosynthetic bacteria strain are respectively activated and cultured on a solid basis, a jelly-like Paenibacillus suspension seed solution with a concentration of 1-3×10 8 CFU/ml is prepared and photosynthetic bacteria suspension seed liquid;
S2-2，将胶冻样类芽孢杆菌菌悬液种液以5％的接种量接种至PDA液体培养基，在摇床上进行单株摇瓶发酵培养，培养温度为35℃，摇床振荡速率为170rpm，培养时间为73h，得到胶冻样类芽孢杆菌菌悬液；S2-2, inoculate the jelly-like Paenibacillus suspension seed liquid into the PDA liquid medium with an inoculum amount of 5%, and carry out a single-plant shake flask fermentation culture on a shaker, the culture temperature is 35°C, and the shaker oscillation rate Be 170rpm, culture time is 73h, obtains jelly-like Paenibacillus suspension;
S2-3，将光合细菌菌悬液种液以5％的接种量接种至RCVBN培养基，在摇床上进行单株摇瓶发酵培养，培养温度为35℃，摇床振荡速率为170rpm，培养时间为73h，得到光合细菌菌悬液；S2-3, inoculate the photosynthetic bacteria suspension seed solution to RCVBN medium with 5% inoculum, carry out single-plant shake flask fermentation culture on a shaker, the culture temperature is 35°C, the shaker oscillation speed is 170rpm, and the culture time For 73h, obtain the photosynthetic bacteria suspension;
S2-4，将7.5kg胶冻样类芽孢杆菌菌悬液和2.5kg光合细菌菌悬液混合加入搅拌锅中，以100r/min的速度匀速搅拌20min，得到复合菌剂B。S2-4, mix 7.5kg of jelly-like Paenibacillus suspension and 2.5kg of photosynthetic bacteria suspension into the stirring pot, stir at a constant speed of 100r/min for 20min, and obtain composite bacterial agent B.
本实施例还公开一种用于甘蔗种植的微生物复合菌剂的制备方法，包括以下步骤：The present embodiment also discloses a preparation method of a microbial composite bacterial agent for sugarcane plantation, comprising the following steps:
取用3.5kg复合菌剂A、5kg复合菌剂B和1.5kg调节剂加入反应釜中，在加热温度为35℃，搅拌速度为500r/min的条件下，每5min进行搅拌一次，加热搅拌30min后取出，静止降温1h，得到微生物复合菌剂。Take 3.5kg of composite bacterial agent A, 5kg of composite bacterial agent B and 1.5kg of regulator and add them to the reaction kettle. Under the condition of heating temperature of 35°C and stirring speed of 500r/min, stir once every 5min, heat and stir for 30min Afterwards, take it out, let it cool down for 1 hour, and obtain the microbial compound bacterial agent.
实施例2Example 2
本实施例公开一种用于甘蔗种植的微生物复合菌剂，包括以下原料：This embodiment discloses a kind of microbial composite bacteria agent for sugarcane plantation, including the following raw materials:
4.0kg复合菌剂A；5.5kg复合菌剂B；0.5kg调节剂；调节剂为乙烯 利。4.0kg composite bacterial agent A; 5.5kg composite bacterial agent B; 0.5kg regulator; regulator is ethylene profit.
复合菌剂A的制备方法为：The preparation method of composite microbial agent A is:
S1-1，将地衣芽孢杆菌和尾孢菌菌株分别固体活化培养后，制得浓度均为1-3×108CFU/ml的地衣芽孢杆菌菌悬液种液和尾孢菌菌悬液种液；S1-1, after solid activation and culture of Bacillus licheniformis and Cercospora strains respectively, Bacillus licheniformis suspension seed liquid and Cercospora bacterial suspension seed liquid were prepared with a concentration of 1-3×10 8 CFU/ml liquid;
S1-2，将地衣芽孢杆菌菌悬液种液以7％的接种量接种至PDA液体培养基，在摇床上进行单株摇瓶发酵培养，培养温度为35℃，摇床振荡速率为170rpm，培养时间为73h，得到地衣芽孢杆菌菌悬液；S1-2, inoculate the Bacillus licheniformis suspension seed solution to the PDA liquid medium with an inoculum amount of 7%, carry out a single-plant shake flask fermentation culture on a shaker, the culture temperature is 35 ° C, and the shaker oscillation speed is 170 rpm, The culture time is 73h, and the suspension of Bacillus licheniformis is obtained;
S1-3，将尾孢菌菌悬液种液以5％的接种量接种至PDA培养基，在摇床上进行单株摇瓶发酵培养，培养温度为35℃，摇床振荡速率为170rpm，培养时间为73h，得到尾孢菌菌悬液；S1-3, inoculate the Cercospora suspension seed solution to the PDA medium with a 5% inoculum amount, carry out single-plant shake flask fermentation culture on a shaker, the culture temperature is 35°C, and the shaker oscillation speed is 170rpm. The time is 73h to obtain the suspension of Cercospora bacteria;
S1-4，将5.5kg地衣芽孢杆菌菌悬液、2.5kg尾孢菌菌悬液和2kg海藻酸混合加入搅拌锅中，以100r/min的速度匀速搅拌20min，得到复合菌剂A；S1-4, mix 5.5kg of Bacillus licheniformis suspension, 2.5kg of Cercospora suspension and 2kg of alginic acid into the stirring pot, and stir at a constant speed of 100r/min for 20min to obtain the composite microbial agent A;
复合菌剂B的制备方法为：The preparation method of composite microbial agent B is:
S2-1，将胶冻样类芽孢杆菌菌种和光合细菌菌种分别固体活化培养后，制得浓度均为1-3×108CFU/ml的胶冻样类芽孢杆菌菌悬液种液和光合细菌菌悬液种液；S2-1, after the jelly-like Paenibacillus strain and the photosynthetic bacteria strain are respectively activated and cultured on a solid basis, a jelly-like Paenibacillus suspension seed solution with a concentration of 1-3×10 8 CFU/ml is prepared and photosynthetic bacteria suspension seed liquid;
S2-2，将胶冻样类芽孢杆菌菌悬液种液以5％的接种量接种至PDA液体培养基，在摇床上进行单株摇瓶发酵培养，培养温度为32℃，摇床振荡速率为150rpm，培养时间为72h，得到胶冻样类芽孢杆菌菌悬液；S2-2, inoculate the jelly-like Paenibacillus suspension seed liquid into the PDA liquid medium with an inoculum amount of 5%, and carry out a single-plant shake flask fermentation culture on a shaker, the culture temperature is 32 ° C, and the shaker shakes at a high rate Be 150rpm, culture time is 72h, obtains jelly-like Paenibacillus suspension;
S2-3，将光合细菌菌悬液种液以3％的接种量接种至RCVBN培养基，在摇床上进行单株摇瓶发酵培养，培养温度为32℃，摇床振荡速率为 150rpm，培养时间为72h，得到光合细菌菌悬液；S2-3, inoculate photosynthetic bacteria suspension seed solution to RCVBN medium with 3% inoculum amount, and carry out single-plant shake flask fermentation culture on a shaker, the culture temperature is 32 ° C, and the shaker shake rate is 150rpm, culture time is 72h, obtains photosynthetic bacteria suspension;
S2-4，将7.5kg胶冻样类芽孢杆菌菌悬液和2.5kg光合细菌菌悬液混合加入搅拌锅中，以100r/min的速度匀速搅拌20min，得到复合菌剂B。S2-4, mix 7.5kg of jelly-like Paenibacillus suspension and 2.5kg of photosynthetic bacteria suspension into the stirring pot, stir at a constant speed of 100r/min for 20min, and obtain composite bacterial agent B.
本实施例还公开一种用于甘蔗种植的微生物复合菌剂的制备方法，包括以下步骤：The present embodiment also discloses a preparation method of a microbial composite bacterial agent for sugarcane plantation, comprising the following steps:
取用4kg复合菌剂A、5.5kg复合菌剂B和0.5kg调节剂加入反应釜中，在加热温度为37℃，搅拌速度为450r/min的条件下，每5min进行搅拌一次，加热搅拌35min后取出，静止降温1h，得到微生物复合菌剂。Take 4kg of composite bacterial agent A, 5.5kg of composite bacterial agent B and 0.5kg of regulator and add them to the reaction kettle. Under the conditions of heating temperature of 37°C and stirring speed of 450r/min, stir once every 5min, heat and stir for 35min Afterwards, take it out, let it cool down for 1 hour, and obtain the microbial compound bacterial agent.
实施例3Example 3
本实施例公开一种用于甘蔗种植的微生物复合菌剂，包括以下原料：This embodiment discloses a kind of microbial composite bacteria agent for sugarcane plantation, including the following raw materials:
3.5kg复合菌剂A；5.0kg复合菌剂B；1.5kg调节剂；调节剂为乙烯利。3.5kg composite bacterial agent A; 5.0kg composite bacterial agent B; 1.5kg regulator; regulator is ethephon.
复合菌剂A包括地衣芽孢杆菌菌悬液60wt％、尾孢菌菌悬液30wt％以及海藻酸10wt％；复合菌剂A的制备方法为：Composite bacterial agent A comprises 60wt% of bacillus licheniformis suspension, 30wt% of cercospora bacterial suspension and 10wt% of alginic acid; the preparation method of composite bacterial agent A is:
S1-1，将地衣芽孢杆菌和尾孢菌菌株分别固体活化培养后，制得浓度均为1-3×108CFU/ml的地衣芽孢杆菌菌悬液种液和尾孢菌菌悬液种液；S1-1, after solid activation and culture of Bacillus licheniformis and Cercospora strains respectively, Bacillus licheniformis suspension seed liquid and Cercospora bacterial suspension seed liquid were prepared with a concentration of 1-3×10 8 CFU/ml liquid;
S1-2，将地衣芽孢杆菌菌悬液种液以5％的接种量接种至PDA液体培养基，在摇床上进行单株摇瓶发酵培养，培养温度为32℃，摇床振荡速率为150rpm，培养时间为72h，得到地衣芽孢杆菌菌悬液；S1-2, inoculate the Bacillus licheniformis suspension seed solution to the PDA liquid medium with an inoculum amount of 5%, carry out a single-plant shake flask fermentation culture on a shaker, the culture temperature is 32 ° C, and the shaker oscillation speed is 150 rpm, The culture time is 72 hours, and the suspension of Bacillus licheniformis is obtained;
S1-3，将尾孢菌菌悬液种液以3％的接种量接种至PDA培养基，在摇床上进行单株摇瓶发酵培养，培养温度为32℃，摇床振荡速率为150rpm，培养时间为72h，得到尾孢菌菌悬液； S1-3, inoculate the Cercospora suspension seed solution to the PDA medium with a 3% inoculum amount, carry out single-plant shake flask fermentation culture on a shaker, the culture temperature is 32°C, and the shaker oscillation speed is 150rpm. The time is 72h to obtain the suspension of Cercospora bacteria;
S1-4，将6.0kg地衣芽孢杆菌菌悬液、3.0kg尾孢菌菌悬液和1kg海藻酸混合加入搅拌锅中，以100r/min的速度匀速搅拌15min，得到复合菌剂A；S1-4, mix 6.0kg of Bacillus licheniformis suspension, 3.0kg of Cercospora suspension and 1kg of alginic acid into the stirring pot, and stir at a constant speed of 100r/min for 15min to obtain composite microbial agent A;
复合菌剂B包括胶冻样类芽孢杆菌菌悬液70wt％和光合细菌菌悬液30wt％。复合菌剂B的制备方法为：Composite microbial agent B includes 70 wt% of jelly-like Paenibacillus suspension and 30 wt% of photosynthetic bacteria suspension. The preparation method of composite microbial agent B is:
S2-1，将胶冻样类芽孢杆菌菌种和光合细菌菌种分别固体活化培养后，制得浓度均为1-3×108CFU/ml的胶冻样类芽孢杆菌菌悬液种液和光合细菌菌悬液种液；S2-1, after the jelly-like Paenibacillus strain and the photosynthetic bacteria strain are respectively activated and cultured on a solid basis, a jelly-like Paenibacillus suspension seed solution with a concentration of 1-3×10 8 CFU/ml is prepared and photosynthetic bacteria suspension seed liquid;
S2-2，将胶冻样类芽孢杆菌菌悬液种液以5％的接种量接种至PDA液体培养基，在摇床上进行单株摇瓶发酵培养，培养温度为35℃，摇床振荡速率为170rpm，培养时间为73h，得到胶冻样类芽孢杆菌菌悬液；S2-2, inoculate the jelly-like Paenibacillus suspension seed liquid into the PDA liquid medium with an inoculum amount of 5%, and carry out a single-plant shake flask fermentation culture on a shaker, the culture temperature is 35°C, and the shaker oscillation rate Be 170rpm, culture time is 73h, obtains jelly-like Paenibacillus suspension;
S2-3，将光合细菌菌悬液种液以5％的接种量接种至RCVBN培养基，在摇床上进行单株摇瓶发酵培养，培养温度为35℃，摇床振荡速率为170rpm，培养时间为73h，得到光合细菌菌悬液；S2-3, inoculate the photosynthetic bacteria suspension seed solution to RCVBN medium with 5% inoculum, carry out single-plant shake flask fermentation culture on a shaker, the culture temperature is 35°C, the shaker oscillation speed is 170rpm, and the culture time For 73h, obtain the photosynthetic bacteria suspension;
S2-4，将7.0kg胶冻样类芽孢杆菌菌悬液和3.0kg光合细菌菌悬液混合加入搅拌锅中，以100r/min的速度匀速搅拌20min，得到复合菌剂B。S2-4, mix 7.0 kg of jelly-like Paenibacillus suspension and 3.0 kg of photosynthetic bacteria suspension into a stirring pot, and stir at a constant speed of 100 r/min for 20 minutes to obtain composite bacterial agent B.
本实施例还公开一种用于甘蔗种植的微生物复合菌剂的制备方法，包括以下步骤：The present embodiment also discloses a preparation method of a microbial composite bacterial agent for sugarcane plantation, comprising the following steps:
取用3.5kg复合菌剂A、5kg复合菌剂B和1.5kg调节剂加入反应釜中，在加热温度为35℃，搅拌速度为500r/min的条件下，每5min进行搅拌一次，加热搅拌30min后取出，静止降温1h，得到微生物复合菌剂。Take 3.5kg of composite bacterial agent A, 5kg of composite bacterial agent B and 1.5kg of regulator and add them to the reaction kettle. Under the condition of heating temperature of 35°C and stirring speed of 500r/min, stir once every 5min, heat and stir for 30min Afterwards, take it out, let it cool down for 1 hour, and obtain the microbial compound bacterial agent.
对比例 comparative example
对比例1Comparative example 1
一种用于甘蔗种植的微生物复合菌剂，与实施例1的不同之处在于：不含有复合菌剂B和调节剂。A microbial compound bacterial agent for sugarcane planting differs from Example 1 in that it does not contain compound bacterial agent B and regulator.
对比例2Comparative example 2
一种用于甘蔗种植的微生物复合菌剂，与实施例1的不同之处在于：不含有复合菌剂A和调节剂。A kind of microbial compound bacterial agent that is used for sugarcane plantation, the difference with embodiment 1 is: not containing compound bacterial agent A and regulator.
对比例3Comparative example 3
一种用于甘蔗种植的微生物复合菌剂，与实施例1的不同之处在于：不含有复合菌剂A和复合菌剂B。A microbial composite bacterial agent for sugarcane planting differs from Example 1 in that it does not contain composite microbial agent A and composite bacterial agent B.
对比例4Comparative example 4
一种用于甘蔗种植的微生物复合菌剂，与实施例1的不同之处在于：不含有调节剂。A microbial compound bacterial agent for sugarcane planting differs from Example 1 in that it does not contain regulators.
对比例5Comparative example 5
一种用于甘蔗种植的微生物复合菌剂，与实施例1的不同之处在于：不含有复合菌剂B。A microbial composite bacterial agent for sugarcane planting, the difference from Example 1 is: no composite bacterial agent B.
对比例6Comparative example 6
一种用于甘蔗种植的微生物复合菌剂，与实施例1的不同之处在于：不含有复合菌剂A。A microbial compound bacterial agent used for sugarcane planting differs from Example 1 in that it does not contain compound bacterial agent A.
对比例7Comparative example 7
一种用于甘蔗种植的微生物复合菌剂，与实施例1的不同之处在于：将地衣芽孢杆菌菌悬液替换为等量的解淀粉芽孢杆菌菌悬液。A kind of microbial composite bacterial agent that is used for sugarcane planting, and the difference of embodiment 1 is: the suspension of Bacillus licheniformis is replaced by the suspension of Bacillus amyloliquefaciens of equal amount.
对比例8 Comparative example 8
一种用于甘蔗种植的微生物复合菌剂，与实施例1的不同之处在于：将尾孢菌菌悬液替换为等量的放线菌菌悬液，放线菌菌悬液的制备方法与尾孢菌菌悬液的制备方法相同，培养基选用PDA培养基。A kind of microbial compound bacterial agent that is used for sugarcane plantation, the difference with embodiment 1 is: the cercospora suspension is replaced by the actinomycetes suspension of equal amount, the preparation method of actinomyces suspension The preparation method is the same as that of the Cercospora suspension, and the medium is PDA medium.
对比例9Comparative example 9
一种用于甘蔗种植的微生物复合菌剂，与实施例1的不同之处在于：将海藻酸替换为等量的黄腐酸。A microbial compound bacterial agent for sugarcane cultivation, the difference from Example 1 is that alginic acid is replaced by an equivalent amount of fulvic acid.
性能检测试验及数据分析Performance testing and data analysis
试验一test one
(1)种植时间：2020年4月-12月(1) Planting time: April-December 2020
(2)种植地点：覃塘区樟木镇邓保村、黄鹤村；(2) Planting locations: Dengbao Village and Huanghe Village, Zhangmu Town, Qintang District;
(3)供试品种：桂糖44号(3) Test variety: Guitang No. 44
(4)方案设计：(4) Scheme design:
1、黄鹤村选取试验田划分为均等的13份，记为1-1、1-2、1-3等；每份种植10株桂糖44号新植；1. The experimental field selected by Huanghe Village is divided into 13 equal parts, which are recorded as 1-1, 1-2, 1-3, etc.; 10 new plants of Guitang No. 44 are planted in each part;
2、邓保村中选取试验田划分为均等的13份，记为2-1、2-2、2-3等；每份种植10株桂糖44号宿根；2. Select the experimental field in Dengbao Village and divide it into 13 equal parts, which are recorded as 2-1, 2-2, 2-3, etc.; plant 10 Guitang No. 44 perennials in each part;
(5)样品：选用市售复混肥125kg+微生物复合菌剂2kg(邓保村试验田和黄鹤村试验田中的一份田地对应选用一个实施例或对比例制得的微生物复合菌剂)其中一份不使用微生物复合菌剂，其他管理措施都一致作为对照组。(5) Sample: select commercially available compound fertilizer 125kg+microbial compound bacterial agent 2kg (a part of the field in the Dengbao Village test field and the Huanghe Village test field corresponds to selecting an embodiment or the microbial compound bacterial agent made in the comparative example) wherein one part does not The use of microbial compound bacterial agents and other management measures are consistent as the control group.
(6)结果：实施例1-3和对比例1-9以及对照组的试验检测数据详见表2(每份试验田的数据取用10株甘蔗的平均值)。 (6) Result: The experimental detection data of embodiment 1-3 and comparative example 1-9 and control group are shown in Table 2 (the average value of 10 sugarcane plants is used for the data of each test field).
表2
Table 2
根据表2中新植和宿根两个试验田甘蔗的性能检测数据可得，新植甘蔗的含糖量和生长活性相较于宿根更加优秀。According to the performance test data of sugarcane in the two test fields of newly planted and perennial roots in Table 2, the sugar content and growth activity of newly planted sugarcane are better than those of perennial roots.
实施例1-3中制得的微生物复合菌剂包含有复合菌剂A、复合菌剂B以及调节剂三大组分，三大组分之间相互促进，共同作用，使得使用了实施例1-3制得的复合菌剂的甘蔗相较于对照组的甘蔗产量更好、含糖量更高，更加接近“双高”甘蔗的种植目标。 The microbial compound microbial agent that makes among the embodiment 1-3 comprises three major components of composite microbial agent A, composite bacterial agent B and regulator, and the mutual promotion between the three major components works together, so that using embodiment 1 Compared with the sugarcane of the control group, the sugarcane of the compound bacterial agent prepared in -3 has better yield and higher sugar content, and is closer to the planting goal of "double high" sugarcane.
对比例1中不含有复合菌剂B和调节剂，对比例1相较于空白对照组多加了复合菌剂A，使用对比例1制得的复合菌剂得到的甘蔗相较于对照得到的甘蔗在株高、茎径、蔗糖份和田间锤度都得到了提升，特别是株高、茎径的提升较为明显，说明复合菌剂A具有促进甘蔗生长的作用；对比例2不含有复合菌剂A和调节剂，对比例2相较于对照组多加了复合菌剂B，使用对比例2制得的复合菌剂得到的甘蔗相较于空白对照组得到的甘蔗在株高、茎径、蔗糖份和田间锤度也都得到了提升，特别是蔗糖份和田间锤度的提升较为明显，说明复合菌剂B具有提升甘蔗蔗糖含量的作用。Composite bacterial agent B and regulator were not contained in comparative example 1, compared with blank control group, comparative example 1 added composite bacterial agent A, and the sugarcane obtained by using the composite bacterial agent obtained in comparative example 1 was compared with the sugarcane obtained by the control The plant height, stem diameter, sucrose content and field hammer have all been improved, especially the improvement of plant height and stem diameter is more obvious, indicating that the compound bacterial agent A has the effect of promoting the growth of sugarcane; comparative example 2 does not contain the compound bacterial agent A and regulator, compared with the control group in comparative example 2, the compound microbial agent B was added more. Compared with the sugarcane obtained by the blank control group, the sugarcane obtained by using the compound bacterial agent obtained in comparative example 2 had higher plant height, stem diameter, sucrose The content and field Brix have also been improved, especially the increase of sucrose content and field Brix is more obvious, indicating that the compound bacterial agent B has the effect of increasing the sucrose content of sugarcane.
对比例4中不含有调节剂，对比例4相较于对照组多加了复合菌剂A和复合菌剂B，使用对比例4制得复合菌剂得到的甘蔗相较于使用对比例1、2得到甘蔗的株高、茎径、蔗糖份和田间锤度的提升较小，说明复合菌剂A和复合菌剂B两者共同使用时可能会产生相互抑制的作用；对比例5是复合菌剂A和调节剂混合、对比例6是复合菌剂B和调节剂混合，对比例5、6的性能检测数据数值同样高于对比例4的性能检测数据数值，进一步反映了复合菌剂A和复合菌剂B两者之间的抑制作用。Comparing example 4 does not contain regulator, comparative example 4 has added composite bacterial agent A and composite bacterial agent B more compared to the control group, and the sugarcane obtained by using comparative example 4 to make composite bacterial agent is compared with using comparative examples 1 and 2 Obtained sugarcane plant height, stem diameter, sucrose content and field Brix have a small increase, indicating that composite bacterial agent A and composite bacterial agent B may produce mutual inhibition when they are used together; comparative example 5 is a composite bacterial agent A is mixed with regulator, and comparative example 6 is that compound bacterial agent B is mixed with regulator, and the performance test data values of comparative examples 5 and 6 are also higher than the performance test data values of comparative example 4, further reflecting compound bacterial agent A and compound bacterial agent Inhibitory effect between the two agents B.
据推测：复合菌剂A中地衣芽孢杆菌、尾孢菌和海藻酸三者共同作用，有助于促进甘蔗的新陈代谢，提升植株的生长效率；而复合菌剂B中胶冻样类芽孢杆菌和光合细菌共同作用，增加了土壤中的氮、钾、磷、硼等多种元素的生成和稳定，有助于促进甘蔗中蔗糖含量的积累；但是由于植株生长需要将积累的蔗糖一部分分解为生长所需的营养物质，故复合菌剂A和复合菌剂B两者一方对蔗糖进行合成、另一方则促进蔗糖分解消耗，如此会产生两者之间相互抑制的情况，调节剂的加入能够与复合菌剂A和复 合菌剂B配合，共同作用下使得体系中蔗糖的积累或转化趋于平衡且进一步促进植株生长和蔗糖的累积，进而达到甘蔗的“高产量、高糖分”的效果。It is speculated that the joint action of Bacillus licheniformis, Cercospora and alginic acid in compound bacterial agent A helps to promote the metabolism of sugarcane and improve the growth efficiency of plants; while the jelly-like Paenibacillus and Photosynthetic bacteria work together to increase the production and stability of nitrogen, potassium, phosphorus, boron and other elements in the soil, which helps to promote the accumulation of sucrose in sugarcane; however, due to plant growth, part of the accumulated sucrose needs to be decomposed into The required nutrients, so one side of compound bacterial agent A and compound bacterial agent B synthesizes sucrose, and the other promotes the decomposition and consumption of sucrose, so that there will be mutual inhibition between the two, and the addition of regulators can be combined with Compound Bacteria A and Complex Combined with Bacteria B, under the combined action, the accumulation or conversion of sucrose in the system tends to be balanced and further promotes plant growth and sucrose accumulation, thereby achieving the effect of "high yield and high sugar content" of sugarcane.
对比例7中将地衣芽孢杆菌替换为等量的解淀粉芽孢杆菌，对比例7制得的复合菌剂对甘蔗株高、茎径的增益作用减弱，说明解淀粉芽孢杆菌、尾孢菌和海藻酸三者之间混合达不到实施例1中地衣芽孢杆菌、尾孢菌和海藻酸三者对甘蔗生长的增益效果；同理，对比例8中将尾孢菌替换为等量的放线菌、对比例9中将海藻酸替换为等量的黄腐酸，使用对比例8、9制得复合菌剂得到的甘蔗的性能检测数据和使用对比例7的甘蔗性能检测数据相近，说明替换地衣芽孢杆菌、尾孢菌和海藻酸三者之间能够起到一定的协同作用，替换后的物质之间无法起到这种协同作用。据推测：地衣芽孢杆菌具有较强的蛋白酶、脂肪酶、淀粉酶的活性，能够促进植株的吸收利用；尾孢菌产生的尾孢菌素对植株的生物活性具有一定的刺激作用，海藻酸可以强化细胞壁，具有稳定的特性，三者之间混合后彼此间相互促进，能够较大范围地刺激植株生长所需酶和激素的活性，使得植株高效生长。In Comparative Example 7, Bacillus licheniformis was replaced with an equivalent amount of Bacillus amyloliquefaciens, and the compound bacterial agent prepared in Comparative Example 7 weakened the effect of gaining sugarcane plant height and stem diameter, indicating that Bacillus amyloliquefaciens, Cercospora and seaweed Mixing between the acid three does not reach the gain effect of Bacillus licheniformis, Cercospora and alginic acid three on sugarcane growth in embodiment 1; Similarly, in comparative example 8, Cercospora is replaced by an equal amount of actinomycetes Alginic acid is replaced by equal amount of fulvic acid in bacteria, comparative example 9, and the performance test data of the sugarcane that uses comparative example 8,9 to make the sugarcane performance test data that composite bacterial agent obtains is similar to the sugarcane performance test data that uses comparative example 7, illustrates replacement Bacillus licheniformis, Cercospora and alginic acid can play a certain synergistic effect, but the replaced substances cannot play this synergistic effect. It is speculated that Bacillus licheniformis has strong protease, lipase, and amylase activities, which can promote the absorption and utilization of plants; the cercosporin produced by Cercospora has a certain stimulating effect on the biological activity of plants, and alginic acid can It strengthens the cell wall and has stable characteristics. After the three are mixed, they promote each other and can stimulate the activity of enzymes and hormones required for plant growth in a large range, so that plants can grow efficiently.
实施例1-3中调节剂选用了乙烯利，使用实施例1-3制得复合菌剂得到的甘蔗产量和含糖量相近，对甘蔗产量和含糖量有良好的增益作用。In Example 1-3, ethephon was selected as the regulator, and the yield and sugar content of sugarcane obtained by using the compound bacterial agent prepared in Example 1-3 were similar, and had a good effect on the yield and sugar content of sugarcane.
据推测：乙烯利可以加速细胞的伸长，对细胞的***也有促进作用。复合菌剂A、复合菌剂B和乙烯利之间能够充分发挥作用，进而使得蔗糖分解和积累处于一个高速转化的平衡状态，进而培育得到的甘蔗的产量和糖分均得到提升。 It is speculated that ethephon can accelerate cell elongation and promote cell division. Composite bacterial agent A, composite bacterial agent B and ethephon can fully play a role, so that the decomposition and accumulation of sucrose are in a high-speed transformation equilibrium state, and then the yield and sugar content of the cultivated sugarcane are improved.
试验二Test two
(1)种植时间：2021年4月-12月；(1) Planting time: April-December 2021;
(2)种植地点：覃塘区樟木镇邓保村、中塘村、黄鹤村；(2) Planting locations: Dengbao Village, Zhongtang Village, Huanghe Village, Zhangmu Town, Qintang District;
(3)供试品种：桂糖44号；(3) Test variety: Guitang No. 44;
(4)方案设计：1、邓保村、中塘村种植桂糖44号宿根；(4) Scheme design: 1. Plant Guitang No. 44 perennial root in Dengbao Village and Zhongtang Village;
2、黄鹤村种植桂糖44号新植；2. The new plant of Guitang No. 44 was planted in Huanghe Village;
(5)样品：对照区—市售复混肥125kg；(5) Sample: control area—commercially available compound fertilizer 125kg;
实验区—市售复混肥125kg+微生物复合菌剂2kg(微生物复合菌剂选用实施例1制得的微生物复合菌剂)；Experimental area—commercially available compound fertilizer 125kg+microbial composite bacterial agent 2kg (microbial composite bacterial agent is selected the microbial composite bacterial agent that embodiment 1 makes);
(6)结果：试验检测数据详见表3；(6) Results: See Table 3 for the test data;
表3
table 3
试验三Trial Three
(1)种植时间：2021年4月-12月；(1) Planting time: April-December 2021;
(2)种植地点：贵港市覃塘区石卡镇双高示范基地；(2) Planting location: Shuanggao Demonstration Base, Shika Town, Qintang District, Guigang City;
(3)品种：桂柳05/136；(3) Variety: Laurel 05/136;
(4)方案设计：均种植宿根；(4) Scheme design: all planted perennial roots;
(5)样品：对照区—种植基地自己配肥； (5) Sample: control area - the planting base mixes its own fertilizer;
实验区—种植基地自己配肥+微生物复合菌剂2kg(微生物复合菌剂选用实施例1制得的微生物复合菌剂)；Experimental area-planting base oneself mixes fertilizer+microbial composite bacterial agent 2kg (microbial composite bacterial agent selects the microbial composite bacterial agent that embodiment 1 makes for use);
(6)结果：试验检测数据详见表4、表5；(6) Results: See Table 4 and Table 5 for the test data;
表4-田间检测数据对比
Table 4 - Comparison of field testing data
表5-糖厂试验室检测数据对比
Table 5-Comparison of test data in sugar factory laboratory
由表4和表5中的数据可知，使用了实施例1制得的微生物复合菌剂的甘蔗相较于未使用微生物复合菌剂的甘蔗在各方面的性能都得到了提升，生长良好，糖分质量较高，大大提升了甘蔗的产量和含糖量，进而可以带来较为可观的经济效益。As can be seen from the data in Table 4 and Table 5, compared with the sugarcane that did not use the microbial composite bacterial agent, the sugarcane that used the microbial composite bacterial agent obtained in Example 1 has improved in all aspects, with good growth and sugar content. The higher quality greatly increases the yield and sugar content of sugarcane, which in turn can bring considerable economic benefits.
以上均为本申请的较佳实施例，并非依此限制本申请的保护范围，故：凡依本申请的结构、形状、原理所做的等效变化，均应涵盖于本申请的保护范围之内。 All of the above are preferred embodiments of the application, and are not intended to limit the protection scope of the application. Therefore, all equivalent changes made according to the structure, shape, and principle of the application should be covered by the protection scope of the application. Inside.

Claims (5)

  1. 一种用于甘蔗种植的微生物复合菌剂，其特征在于：包括以下质量百分比的组分：A microbial compound bacterial agent for planting sugarcane is characterized in that: it comprises the following components by mass percentage:
    复合菌剂A 35-40wt％；Composite bacterial agent A 35-40wt%;
    复合菌剂B 50-55wt％；Composite bacterial agent B 50-55wt%;
    调节剂5-15wt％；Regulator 5-15wt%;
    所述复合菌剂A包括55-60wt％的地衣芽孢杆菌菌悬液、25-30wt％的尾孢菌菌悬液以及10-20wt％的海藻酸；The composite bacterial agent A includes 55-60wt% of Bacillus licheniformis suspension, 25-30wt% of Cercospora suspension and 10-20wt% of alginic acid;
    所述复合菌剂A的制备方法包括：The preparation method of described composite bacterial agent A comprises:
    S1-1，将地衣芽孢杆菌和尾孢菌菌株分别固体活化培养后，制得浓度均为1-3×108CFU/ml的地衣芽孢杆菌菌悬液种液和尾孢菌菌悬液种液；S1-1, after solid activation and culture of Bacillus licheniformis and Cercospora strains respectively, Bacillus licheniformis suspension seed liquid and Cercospora bacterial suspension seed liquid were prepared with a concentration of 1-3×10 8 CFU/ml liquid;
    S1-2，将地衣芽孢杆菌菌悬液种液以5-7％的接种量接种至培养基，培养温度为32-35℃，培养时间为72-76h，150-170rpm振荡培养得到地衣芽孢杆菌菌悬液；S1-2, inoculate the Bacillus licheniformis suspension seed solution to the culture medium with an inoculum amount of 5-7%, the culture temperature is 32-35°C, the culture time is 72-76h, and 150-170rpm vibration culture is obtained to obtain Bacillus licheniformis bacterial suspension;
    S1-3，将尾孢菌菌悬液种液以3-5％的接种量接种至培养基，培养温度为32-35℃，培养时间为72-76h，150-170rpm振荡培养得到尾孢菌菌悬液；S1-3, inoculate the Cercospora suspension seed solution to the culture medium with an inoculum size of 3-5%, the culture temperature is 32-35°C, the culture time is 72-76h, and the Cercospora is obtained by shaking culture at 150-170rpm bacterial suspension;
    S1-4，将地衣芽孢杆菌菌悬液、尾孢菌菌悬液和海藻酸混合，得到复合菌剂A；S1-4, mixing Bacillus licheniformis suspension, Cercospora suspension and alginic acid to obtain composite microbial agent A;
    所述复合菌剂B包括25-30wt％的光合细菌菌悬液以及胶冻样类芽孢杆菌菌悬液余量；The composite bacterial agent B includes 25-30wt% photosynthetic bacteria suspension and jelly-like Paenibacillus suspension balance;
    所述复合菌剂B的制备方法包括：The preparation method of described composite bacterial agent B comprises:
    S2-1，将胶冻样类芽孢杆菌菌种和光合细菌菌种分别固体活化培养后，制得浓度均为1-3×108CFU/ml的胶冻样类芽孢杆菌菌悬液种液和光合细菌菌悬液种液；S2-1, after the jelly-like Paenibacillus strain and the photosynthetic bacteria strain are respectively activated and cultured on a solid basis, a jelly-like Paenibacillus suspension seed solution with a concentration of 1-3×10 8 CFU/ml is prepared and photosynthetic bacteria suspension seed liquid;
    S2-2，将胶冻样类芽孢杆菌菌悬液种液以5-7％的接种量接种至培养基，培养温度为32-35℃，培养时间为72-76h，150-170rpm振荡培养得到胶冻样类芽孢杆菌菌悬液；S2-2, inoculate the jelly-like Paenibacillus suspension seed liquid into the culture medium with an inoculum size of 5-7%, the culture temperature is 32-35°C, the culture time is 72-76h, and the culture medium is shaken at 150-170rpm to obtain Jelly-like Paenibacillus suspension;
    S2-3，将光合细菌菌悬液种液以3-5％的接种量接种至培养基，培养温度为32-35℃，培养时间为72-76h，150-170rpm振荡培养得到光合细菌菌悬液；S2-3, inoculate the seed solution of the photosynthetic bacteria suspension to the culture medium with an inoculum size of 3-5%, the culture temperature is 32-35°C, the culture time is 72-76h, and the photosynthetic bacteria suspension is obtained by oscillating culture at 150-170rpm. liquid;
    S2-4，将胶冻样类芽孢杆菌菌悬液和光合细菌菌悬液混合,得到复合菌剂B。S2-4, mixing the jelly-like Paenibacillus suspension and the photosynthetic bacteria suspension to obtain a composite bacterial agent B.
  2. 根据权利要求1所述的一种用于甘蔗种植的微生物复合菌剂，其特征在于：所述复合菌剂A包括以下质量百分比的组分：57-58wt％的地衣芽孢杆菌菌悬液、27-28wt％的尾孢菌菌悬液以及14-16wt％的海藻酸；所述复合菌剂B包括以下质量百分比的组分：25-28wt％的光合细菌菌悬液以及胶冻样类芽孢杆菌菌悬液余量。A microbial composite bacterial agent for sugarcane plantation according to claim 1, characterized in that: said composite microbial agent A comprises the following components in mass percent: 57-58wt% of Bacillus licheniformis suspension, 27 -28wt% of Cercospora suspension and 14-16wt% of alginic acid; the composite bacterial agent B includes the following components in mass percentage: 25-28wt% of photosynthetic bacteria suspension and jelly-like Paenibacillus The remaining amount of bacterial suspension.
  3. 根据权利要求2所述的一种用于甘蔗种植的微生物复合菌剂，其特征在于：所述复合菌剂A包括以下质量百分比的组分：57.5wt％的地衣芽孢杆菌菌悬液、27.5wt％的尾孢菌菌悬液以及15wt％的海藻酸；所述复合菌剂B至少包括以下质量百分比的组分：26.5wt％的光合细菌菌悬液以及胶冻样类芽孢杆菌菌悬液余量。A kind of microbial composite bacterial agent for planting sugarcane according to claim 2, is characterized in that: described composite bacterial agent A comprises the following components of mass percent: 57.5wt% Bacillus licheniformis bacterial suspension, 27.5wt% % of Cercospora bacteria suspension and 15wt% alginic acid; the composite bacterial agent B includes at least the following mass percentage components: 26.5wt% of photosynthetic bacteria suspension and jelly-like Paenibacillus suspension quantity.
  4. 根据权利要求1-3任一所述的一种用于甘蔗种植的微生物复合菌剂，其特征在于：所述调节剂为乙烯利。A microbial compound bacterial agent for sugarcane planting according to any one of claims 1-3, characterized in that: the regulator is ethephon.
  5. [根据细则26改正 07.04.2023]
    一种如权利要求1-4任一所述的用于甘蔗种植的微生物复合菌剂的制备方法，其特征在于：包括以下步骤：将复合菌剂A、复合菌剂B和调节剂混合均匀，得到微生物复合菌剂。    [Corrected 07.04.2023 under Rule 26]
    A preparation method for the microbial composite bacterial agent for sugarcane plantation as described in any one of claims 1-4, it is characterized in that: comprise the following steps: mixing composite bacterial agent A, composite bacterial agent B and regulator, Obtain microbial composite bacteria agent.
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