Microalgae biological synergist as well as preparation method and application thereof

Abstract

The invention provides a microalgae biological synergist as well as a preparation method and application thereof, and belongs to the technical field of soil improvement and crop planting. The microalgae biological synergist is prepared from the following raw materials in percentage by weight: 35 to 50 percent of microalgae concentrated solution, 35 to 50 percent of liquid mineral source potassium fulvate and 15 to 30 percent of powdery bacteria, the microalgae are chlorella sorokiniana, and the cell density of the microalgae is (2.5 to 3.5) * 10 < 7 > cells / mL; the powdery bacterium is a composite functional flora composed of bacillus polymyxa, bacillus subtilis, bacillus cereus and bacillus licheniformis, and the effective viable count is greater than or equal to 2.0 billion / g. The biological synergist provided by the invention can effectively improve soil, and can greatly improve the drought resistance, water and fertilizer utilization efficiency, yield, quality and the like of crops.

Description

Technical Field

[0001] The invention belongs to the technical field of soil improvement and crop planting, and in particular relates to a microalgae biosynergist and a preparation method and application thereof. Background Art

[0002] Farmland soil is a precious resource for human survival and an irreplaceable production factor for human agricultural planting. The protection, restoration, improvement, utilization and development of farmland soil are related to the healthy and sustainable development of my country's agriculture, national food security and the physical nutrition and health of the people. Farmland soil improvement and restoration is an important technical means to stick to my country's 1.8 billion mu of arable land red line. my country has a large population, but arable land resources are scarce. The per capita arable land area is only about 40% of the world average. Due to the national conditions of small land and large population, my country's arable land is in a state of high input, high output and overload utilization all year round. The quality of farmland soil has declined, and agricultural stable production, increased production and the quality of agricultural products have been affected to a certain extent. The pollutants accumulated in the soil will pollute the farmland soil and affect food safety. Therefore, it is urgent to do a good job in improving and restoring farmland soil and comprehensively improve the quality of my country's arable land.

[0003] The main problems with farmland soil quality in my country are farmland soil degradation and poor soil fertility; farmland soil pollution and difficulty in management; the current situation of over-utilization of arable land is difficult to fundamentally change, and farmland soil quality continues to deteriorate; companies are flocking into the soil remediation and management industry, and product technologies vary greatly.

[0004] As people pay more and more attention to the research of marine biological resources, the exploration of algae biological resources has also received more and more attention. In recent years, its application and value in the agricultural field have become increasingly prominent. At present, people's research on the application of algae resources in agriculture mainly focuses on the exploration of algae biofertilizers and biostimulants, that is, to find new fertilizer sources for crops.

[0005] As an important part of the biosphere, algae grow widely in water and terrestrial environments, with morphological diversity and functional diversity. Studies have shown that algae cells are widely used in the agricultural field because they are rich in bioactive substances. Algae cells increase the oxygen and organic matter content of the soil through photosynthesis and nitrogen fixation, thereby increasing the activity of soil microorganisms and improving the interaction between microorganisms, promoting plant growth; biostimulants extracted from algae cells can promote seed germination, plant growth, improve plant nutrient utilization efficiency, and enhance plant tolerance to environmental stress; seaweed polysaccharides extracted from algae cells can form biofilms on the surface of agricultural products, inhibit microbial infection and biological oxidation, and are potential preservatives for agricultural products; in terms of soil remediation and pollution control, the extracellular polymers of microalgae can also be used to enhance the stability of soil aggregates and achieve the control of desertified soil; microalgae cells are a natural and efficient biosorbent that can tolerate and enrich metal ions in the environment, thereby achieving the purpose of controlling soil heavy metal pollution; when treating saline-alkali soil, cyanobacteria that produce extracellular polysaccharides can also be used to increase the nitrogen content of saline-alkali soil and alleviate the problem of excessive sodium ion content in saline-alkali soil.

[0006] However, there are very few studies on promoting soil probiotics through algal cell secretions, and few scholars have used desert algae secretions to increase the growth of soil probiotics to achieve the purpose of soil improvement. Summary of the invention

[0007] In order to solve the above technical problems, the present invention provides a microalgae biosynergist and its preparation method and application. The microalgae biosynergist in the present invention comprises the following raw materials in weight percentage: 35-50% microalgae concentrate, 35-50% liquid mineral potassium humate and 15-30% powdery bacteria; the microalgae is Chlorella sorokiniana, with a cell density of 2.5-3.5×10 7 The powdery bacteria are a composite functional bacterial community composed of Bacillus polymyxa, Bacillus subtilis, Bacillus cereus and Bacillus licheniformis, and the effective live bacteria count is ≥ 200 million / gram. The biological enhancer in the present invention can effectively improve the soil and greatly improve the drought resistance, water and fertilizer utilization efficiency, yield and quality of crops.

[0008] In order to achieve the above object, the present invention adopts the following technical solutions:

[0009] The present invention provides a microalgae biosynergist, which comprises the following raw materials in weight percentage: 35-50% of microalgae concentrate, 35-50% of liquid mineral potassium humate and 15-30% of powdery bacteria; the microalgae is Chlorella sorokiniana, and the cell density is 2.5-3.5×10 7The powdery bacteria is a composite functional bacterial community composed of Bacillus polymyxa, Bacillus subtilis, Bacillus cereus and Bacillus licheniformis, and the effective live bacteria count is ≥ 200 million / gram.

[0010] Furthermore, the microalgae biosynergist comprises the following raw materials in percentage by weight: 50% microalgae concentrate, 35% liquid mineral potassium humate and 15% powdered bacteria.

[0011] Furthermore, the cell density of Chlorella sorokinensis in the microalgae biosynergist is 3×10 7 Pieces / mL.

[0012] The present invention also provides a method for preparing the microalgae biosynergist, which specifically comprises the following steps:

[0013] Step 1: preparing microalgae concentrate: (1) preparing a modified BG11 liquid culture medium, and packaging, sealing, and sterilizing at high temperature and high pressure; (2) inoculating a single colony of microalgae in the modified BG11 liquid culture medium, irradiating with light, and culturing statically for 7 to 10 days to obtain microalgae mother cells; (3) taking 1 to 3 ml of culture from the microalgae mother cells, and culturing in the modified BG11 liquid culture medium until the cell density reaches 0.8 to 1.2 × 10 6 cell / mL to obtain the first-level microalgae seed; (4) expanding the first-level microalgae seed cells by 10 times and inoculating them again in the improved BG11 liquid culture medium, illuminating and statically culturing for 7 to 10 days until the cell density reaches 0.8 to 1.2×10 7 cell / mL to obtain microalgae secondary seeds; (5) performing solid-liquid separation on the culture solution of the microalgae secondary seeds, and concentrating the solid part until the cell density reaches 0.8 to 1.2×10 8 cell / mL, and the microalgae concentrate is obtained;

[0014] Step 2: Mix the liquid mineral potassium humate solution and the powdered bacteria, and stir at 25-35° C. for 10-20 minutes;

[0015] Step 3, mixing the microalgae concentrate with the product obtained in step 2, and stirring at 25-30° C. for 20-40 minutes;

[0016] Step 4: Filling and sealing the product obtained in step 3 to obtain the microalgae bio-enhancer.

[0017] Furthermore, the modified BG11 liquid culture medium in step (1) is prepared by adding 1.5 g / L NaNO 3 Change to 5g / L CO(NH 2 ) 2 The conditions for high temperature and high pressure sterilization treatment are: temperature is 121°C, pressure is 0.12MPa, and treatment time is 20min.

[0018] Furthermore, in step (3), the cells are cultured until the cell density reaches 1×10 6 cell / mL; in step (4), the cell density is cultured until 1×10 7 cell / mL; in step (5), the cells are cultured until the cell density reaches 1×10 8 cell / mL.

[0019] Furthermore, the culture conditions in step (2) and step (4) are: temperature of 27-29° C. and light of 3000 lux.

[0020] The present invention also provides an application of the microalgae biosynergist in soil improvement.

[0021] The present invention also provides an application of the microalgae biosynergist in crop planting.

[0022] Compared with the prior art, the present invention has the following technical effects:

[0023] (1) The microalgae biosynergist prepared by the present invention provides a more excellent plant cultivation method, which can effectively improve the activity of soil microbial flora, promote and stimulate the recovery of original microorganisms and soil animals in the soil, and greatly improve the drought resistance, water and fertilizer utilization efficiency, yield and quality of crops.

[0024] (2) The microalgae biostimulant obtained by the present invention is simpler to prepare and easier to operate than conventional microalgae biostimulants. DETAILED DESCRIPTION

[0025] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Without departing from the spirit and essence of the present invention, modifications or replacements made to the method, steps or conditions of the present invention are within the scope of the present invention. In the examples of the present invention, the equipment, raw materials, etc. used are commercially available unless otherwise specified, and the methods used are consistent with conventional methods unless otherwise specified.

[0026] The microalgae used in the present invention is Chlorella sorokiniana; the liquid mineral potassium humate used is produced by Xinjiang Black Ecological Technology Co., Ltd., and is a high-molecular non-uniform aromatic hydroxycarboxylic acid potassium salt liquid, containing humic acid ≥12.0%, fulvic acid ≥10.0%, potassium oxide ≥2.0%, and solid content ≥20.0%; the powdered bacteria used are Bacillus produced by Beijing Zhongnong Xinke Biotechnology Co., Ltd., and are a composite functional bacterial community composed of Bacillus polymyxa, Bacillus subtilis, Bacillus cereus, and Bacillus licheniformis, and the registration certificate number is Microbial Fertilizer (2003) Standard No. (0106), and the number of effective live bacteria is ≥200 million / gram.

[0027] The technical solution of the present invention is further described in detail below in conjunction with embodiments.

[0028] Example 1

[0029] A microalgae biosynergist comprises 35% of microalgae concentrate, 45% of liquid mineral potassium humate and 20% of powdered bacteria, and the preparation method thereof is as follows:

[0030] 1. Preparation of microalgae culture medium: (1) Add 1.5 g / L NaNO 3 Change to 5g / L CO(NH 2 ) 2， Prepare a modified BG11 liquid culture medium and dispense it into 100 mL of a 150 mL Erlenmeyer flask, seal it with a 6×6 cm sealing film, tie it with a high temperature resistant rubber band, and sterilize it at high temperature and high pressure for 20 min at a temperature of 121°C to obtain a liquid culture medium for microalgae cell culture; (2) Use an inoculation loop to take an appropriate amount of single algae colonies under sterile conditions and inoculate them into the prepared liquid culture medium, and illuminate and statically culture it at a temperature of 27 to 29°C and a light intensity of 3000 lux for 7 to 10 days to obtain microalgae mother cells; (3) After the culture is completed, take 2 ml of the culture from the mother cells, observe the cell morphology, cell size, purity, cell number and other indicators under an optical microscope, and culture it until the cell density reaches 1×10 6cell / mL to obtain the first-level microalgae seed; (4) the first-level seed cells were expanded 10 times and inoculated into the prepared liquid culture medium, and the temperature was 27-29°C and the light intensity was 3000 lux. The culture was carried out under illumination and static culture for 7-10 days. During the culture process, 2 ml of sample was taken from the culture medium regularly every day, and the cell morphology, cell size, purity, cell number and other indicators were observed under an optical microscope. The culture was continued until the cell density reached 1×10 7 cell / mL to obtain microalgae secondary seeds; (5) after the culture, the culture solution was separated into solid and liquid by centrifugation, and the obtained solid was concentrated to a cell density of 1×10 8 cell / mL is put into a stirring tank to prepare a microalgae concentrate. 2. Mix the liquid mineral potassium humate solution and the powdered bacteria, place them at 25-35°C and stir for 10-20 minutes; 3. Mix the microalgae concentrate with the product obtained in step 2, place them at 25-30°C and stir for 20-40 minutes; 4. Fill and seal the product obtained in step 3 to obtain a microalgae bio-enhancer.

[0031] Example 2

[0032] A microalgae biosynergist comprises 40% of microalgae concentrate, 45% of liquid mineral potassium humate and 15% of powdered bacteria, and the preparation method thereof is as follows:

[0033] 1. Preparation of microalgae culture medium: (1) Add 1.5 g / L NaNO 3 Change to 5g / L CO(NH 2 ) 2， Prepare a modified BG11 liquid culture medium and dispense it into 100 mL of a 150 mL Erlenmeyer flask, seal it with a 6×6 cm sealing film, tie it with a high temperature resistant rubber band, and sterilize it at high temperature and high pressure for 20 min at a temperature of 121°C to obtain a liquid culture medium for microalgae cell culture; (2) Use an inoculation loop to take an appropriate amount of single algae colonies under sterile conditions and inoculate them into the prepared liquid culture medium, and illuminate and statically culture it at a temperature of 27 to 29°C and a light intensity of 3000 lux for 7 to 10 days to obtain microalgae mother cells; (3) After the culture is completed, take 2 ml of the culture from the mother cells, observe the cell morphology, cell size, purity, cell number and other indicators under an optical microscope, and culture it until the cell density reaches 1×10 6cell / mL to obtain the first-level microalgae seed; (4) the first-level seed cells were expanded 10 times and inoculated into the prepared liquid culture medium, and the temperature was 27-29°C and the light intensity was 3000 lux. The culture was carried out under illumination and static culture for 7-10 days. During the culture process, 2 ml of sample was taken from the culture medium regularly every day, and the cell morphology, cell size, purity, cell number and other indicators were observed under an optical microscope. The culture was continued until the cell density reached 1×10 7 cell / mL to obtain microalgae secondary seeds; (5) after the culture, the culture solution was separated into solid and liquid by centrifugation, and the obtained solid was concentrated to a cell density of 1×10 8 cell / mL is put into a stirring tank to prepare a microalgae concentrate. 2. Mix the liquid mineral potassium humate solution and the powdered bacteria, place them at 25-35°C and stir for 10-20 minutes; 3. Mix the microalgae concentrate with the product obtained in step 2, place them at 25-30°C and stir for 20-40 minutes; 4. Fill and seal the product obtained in step 3 to obtain a microalgae bio-enhancer.

[0034] Example 3

[0035] A microalgae biosynergist comprises 50% of microalgae concentrate, 35% of liquid mineral potassium humate and 15% of powdered bacteria, and the preparation method thereof is as follows:

[0036] 1. Preparation of microalgae culture medium: (1) Add 1.5 g / L NaNO 3 Change to 5g / L CO(NH 2 ) 2， Prepare a modified BG11 liquid culture medium and dispense it into 100 mL of a 150 mL Erlenmeyer flask, seal it with a 6×6 cm sealing film, tie it with a high temperature resistant rubber band, and sterilize it at high temperature and high pressure for 20 min at a temperature of 121°C to obtain a liquid culture medium for microalgae cell culture; (2) Use an inoculation loop to take an appropriate amount of single algae colonies under sterile conditions and inoculate them into the prepared liquid culture medium, and illuminate and statically culture it at a temperature of 27 to 29°C and a light intensity of 3000 lux for 7 to 10 days to obtain microalgae mother cells; (3) After the culture is completed, take 2 ml of the culture from the mother cells, observe the cell morphology, cell size, purity, cell number and other indicators under an optical microscope, and culture it until the cell density reaches 1×10 6cell / mL to obtain the first-level microalgae seed; (4) the first-level seed cells were expanded 10 times and inoculated into the prepared liquid culture medium, and the temperature was 27-29°C and the light intensity was 3000 lux. The culture was carried out under illumination and static culture for 7-10 days. During the culture process, 2 ml of sample was taken from the culture medium regularly every day, and the cell morphology, cell size, purity, cell number and other indicators were observed under an optical microscope. The culture was continued until the cell density reached 1×10 7 cell / mL to obtain microalgae secondary seeds; (5) after the culture, the culture solution was separated into solid and liquid by centrifugation, and the obtained solid was concentrated to a cell density of 1×10 8 cell / mL is put into a stirring tank to prepare a microalgae concentrate. 2. Mix the liquid mineral potassium humate solution and the powdered bacteria, place them at 25-35°C and stir for 10-20 minutes; 3. Mix the microalgae concentrate with the product obtained in step 2, place them at 25-30°C and stir for 20-40 minutes; 4. Fill and seal the product obtained in step 3 to obtain a microalgae bio-enhancer.

[0037] Example 4

[0038] The microalgae biosynergists prepared in Examples 1 to 3 were applied to improve the greenhouse soil in Xishan Farm, Urumqi. Three dosage groups were set up, and the application rate of the high-dose group was 10 L / m 2 The dosage of the medium dose group was 5L / m 2 The low-dose group was administered at a volume of 2.5 L / m 2 ; At the same time, a "bacteria bun" treatment group (co-produced by Shandong Zhengdao Biotechnology Co., Ltd. and Zhucheng Land Organic Fertilizer Co., Ltd.) was set up, with an application rate of 5L / mu; a potassium humate solution + powdered bacteria treatment group, with an application rate of 5L / mu; and a control group without any product. The test plants were tomato and cucumber varieties "Super 100" and "Dinghao Youliang 777" respectively, and the specific cultivation and management methods were consistent with conventional methods.

[0039] (1) The soil pH, salinity, electrical conductivity, available phosphorus, available potassium and ammonium nitrogen in each treatment group were tested. The results are shown in Table 1.

[0040] Table 1 Soil index data of each treatment group

[0041] project Before applying the enhancer First application of booster Second application of booster Salt content (g / L) 0.65 0.59 0.49 Conductivity (mS / cm) 1.59 1.42 0.75 Available phosphorus (mg / kg) 13.8 81.29 82.26 Available potassium (mg / kg) 108.2 273.87 279.95 Ammonium nitrogen (mg / kg) 44.69 65.81 81.46

[0042] The results showed that after the application of microalgae enhancer, the salt content of the soil decreased from 0.65g / L to 0.49g / L, with a decrease rate of 24.61%; the electrical conductivity decreased from 1.59mS / cm to 0.75mS / cm, with a decrease rate of 52.83%; the effective phosphorus content increased from 13.8mg / kg to 82.26mg / kg, with an increase rate of 496%; the fast-acting potassium content increased from 108.2mg / kg to 279.95mg / kg, with an increase rate of 158.73%; the ammonium nitrogen content increased from 44.69mg / kg to 81.46mg / kg, with an increase rate of 82.27%.

[0043] (2) The yields of cucumber and tomato in each treatment group are shown in Table 2.

[0044] Table 2 Yield of cucumber and tomato in each treatment group

[0045] Treatment group (tomato) Output(kg) Treatment group (cucumber) Output(kg) Blank Group 8.384 Blank Group 23.922 Positive control group 10.882 Positive control group 28.451 Microalgae liquid low dose group 10.596 Microalgae liquid low dose group 26.551 Microalgae medium dose group 13.471 Microalgae liquid medium dose group 32.484 Microalgae liquid high dose group 13.643 Microalgae liquid high dose group 33.012 Microalgae enhancer group 14.193 Microalgae enhancer group 33.102 Potassium humate solution + powdered bacteria group 12.665 Potassium humate solution + powdered bacteria group 29.079

[0046] The results showed that the yields of cucumber and tomato were increased by 16.35% and 30.43% respectively in the treatment group using the microalgae synergist of the present invention.

[0047] (3) The quality of cucumbers and tomatoes in the “bacteria bun” treatment group (bio-organic fertilizer group) and the microalgae enhancer treatment group are shown in Table 3.

[0048] Table 3 Quality of cucumber and tomato in each treatment group

[0049]

[0050]

[0051] The quality results showed that the protein and soluble sugar content of cucumber and pepper in the high-dose microalgae treatment group increased by 15.69% and 37.78% respectively. The comparison ratio of potassium fulvic acid solution + bacteria was 35%:15%.

[0052] The above experimental results fully demonstrate that the microalgae biosynergist in the present invention not only has the effect of increasing the yield of greenhouse vegetables, but also has the effect of improving the quality of fruits.

[0053] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A microalgae biosynergist, characterized in that: The microalgae biosynergist comprises the following raw materials in weight percentage: 35-50% of microalgae concentrate, 35-50% of liquid mineral potassium humate and 15-30% of powdered bacteria; the microalgae is Chlorella sorokiniana, with a cell density of 2.5-3.5×10 7 The powdery bacteria is a composite functional bacterial community composed of Bacillus polymyxa, Bacillus subtilis, Bacillus cereus and Bacillus licheniformis, and the effective live bacteria count is ≥ 200 million / gram.

2. The microalgae biostimulant according to claim 1, characterized in that: The microalgae biosynergist comprises the following raw materials in percentage by weight: 50% of microalgae concentrate, 35% of liquid mineral potassium humate and 15% of powdered bacteria.

3. The microalgae biostimulant according to claim 1, characterized in that: The cell density of Chlorella sorokinensis in the microalgae biosynergist is 3×10 7 Pieces / mL.

4. The method for preparing the microalgae biosynergist according to any one of claims 1 to 3, characterized in that: The following steps are involved: Step 1: preparing microalgae concentrate: (1) preparing a modified BG11 liquid culture medium, and packaging, sealing, and sterilizing at high temperature and high pressure; (2) inoculating a single colony of microalgae in the modified BG11 liquid culture medium, irradiating with light, and culturing statically for 7 to 10 days to obtain microalgae mother cells; (3) taking 1 to 3 ml of culture from the microalgae mother cells, and culturing in the modified BG11 liquid culture medium until the cell density reaches 0.8 to 1.2 × 10 6 cell / mL to obtain the first-level microalgae seed; (4) expanding the first-level microalgae seed cells by 10 times and inoculating them again in the improved BG11 liquid culture medium, illuminating and statically culturing for 7 to 10 days until the cell density reaches 0.8 to 1.2×10 7 cell / mL to obtain microalgae secondary seeds; (5) performing solid-liquid separation on the culture solution of the microalgae secondary seeds, and concentrating the solid part until the cell density reaches 0.8 to 1.2×10 8 cell / mL, and the microalgae concentrate is obtained; Step 2: Mix the liquid mineral potassium humate solution and the powdered bacteria, and stir at 25-35° C. for 10-20 minutes; Step 3, mixing the microalgae concentrate with the product obtained in step 2, and stirring at 25-30° C. for 20-40 minutes; Step 4: Filling and sealing the product obtained in step 3 to obtain the microalgae bio-enhancer.

5. The preparation method according to claim 4, characterized in that: The preparation method of the improved BG11 liquid culture medium in step (1) is as follows: replacing 1.5 g / L NaNO3 in the BG11 culture medium with 5 g / L CO(NH2)2; the conditions for high temperature and high pressure sterilization treatment are as follows: temperature of 121°C, pressure of 0.12 MPa, and treatment time of 20 min.

6. The preparation method according to claim 5, characterized in that: In step (3), the cells are cultured until the cell density reaches 1×10 6 cell / mL; in step (4), the cell density is cultured until 1×10 7 cell / mL; in step (5), the cells are cultured until the cell density reaches 1×10 8 cell / mL.

7. The preparation method according to claim 6, characterized in that: The culture conditions in step (2) and step (4) are: temperature of 27-29° C. and light of 3000 lux.

8. Use of the microalgae biostimulant according to any one of claims 1 to 3 or the microalgae biostimulant prepared by the preparation method according to any one of claims 4 to 7 in soil improvement.

9. Use of the microalgae biosynergist according to any one of claims 1 to 3 or the microalgae biosynergist prepared by the preparation method according to any one of claims 4 to 7 in crop planting.

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