Microalgae biological fertilizer as well as preparation method and application thereof

Abstract

The invention relates to the technical field of microbial fertilizers, in particular to a microalgae biological fertilizer and a preparation method and application thereof, and the method comprises the following steps: step 1, algae liquid inoculation, step 2, graded propagation cultivation, step 3, algae cell screening, step 4, compounding and mixing, and step 5, finished product preparation. According to the invention, the microalgae such as the terriella variabilis, the small monofidus algae and the chlorella pyrenoidosa are adopted and combined with the microalgae biological fertilizer prepared by the preparation method disclosed by the invention, so that the competition of the microalgae for nutrients can be reduced, meanwhile, the nutrients can be better produced, and the growth of crops is facilitated.

Description

Technical Field

[0001] The invention relates to the technical field of microbial fertilizers, and in particular to a microalgae biofertilizer and a preparation method and application thereof. Background Art

[0002] In agricultural production, in order to improve the yield and quality of crops, various fertilizers are often used to supplement the nutrients required by plants. Although traditional chemical fertilizers can quickly provide the nutrients required by plants, their pollution to the environment and potential harm to human health have attracted widespread attention. Therefore, in recent years, people have begun to study biofertilizers prepared from microbial resources to achieve green, environmentally friendly and sustainable agricultural production.

[0003] Microalgae are a type of microorganism that grows fast, has a high metabolic intensity, and is rich in bioactive substances such as proteins, lipids, and nucleic acids. They are ideal materials for preparing biofertilizers and can better promote soil health and plant growth. For example, microalgae with nitrogen-fixing effects can provide plants with the nutrient element nitrogen, and spherical algae can release oxygen through photosynthesis.

[0004] However, microalgae are also plants and need to absorb nutrients. Therefore, after being applied to the soil, there is a problem of competing with plants for nutrients, which results in the consumption of some nutrients while producing them.

[0005] Currently, there are no products or methods that can reduce the competition for nutrients from microalgae while better producing nutrients and more conducive to crop growth. Summary of the invention

[0006] The purpose of the present invention is to overcome the shortcomings of the prior art and propose a microalgae biofertilizer and a preparation method and application thereof to solve the problem of reducing the competition of microalgae for nutrients while better producing nutrients, which is beneficial to crop growth.

[0007] The technical solution adopted by the present invention to solve the technical problem is: A microalgae biofertilizer comprises micromonad, pyrenoid chlorella and variegated chlorella, wherein the proportion of variegated chlorella is greater than that of pyrenoid chlorella.

[0008] A method for preparing microalgae biofertilizer comprises the following steps: Step 1: Algal liquid inoculation Inoculate the microalgae into the culture medium and control the temperature, light intensity and pH value; Step 2: Gradual propagation and cultivation The algae species are cultured in stages. When the microalgae cells in the previous stage are ≧1.0*10 6 When the number of cells / ml is reached, the next level of culture is carried out; Step 3: Algal cell screening Use a microscope to test the algae cells to determine whether they are qualified; Unqualified algae cells are inactivated by high temperature treatment and discarded, while qualified algae cells are preserved; Step 4: Preparation of microalgae biofertilizer The preserved qualified algae cells are pumped into a mixing tank through a pipeline for compounding and mixing of algae species to obtain liquid microalgae biofertilizer; Freeze-drying the liquid microalgae biofertilizer to obtain powdered microalgae biofertilizer; The freeze-drying conditions are -20°C to -10°C and the pressure is 100 to 500 Pa; Microalgae biofertilizer includes liquid microalgae biofertilizer and powder microalgae biofertilizer.

[0009] In step 1, the volume ratio of the algae solution to the culture medium in step 1 is 1-5:20-30; The temperature was controlled at 20-30°C, the light intensity was 2000-5000 Lux, and the pH value was 6.5-7.5; The culture medium comprises the following components by weight: 1-5 parts of sodium bicarbonate, 1-5 parts of sodium nitrate, 0.8-2 parts of potassium dihydrogen phosphate, 0.01-0.05 parts of calcium disodium ethylenediaminetetraacetate, 0.01-0.05 parts of trace elements, and 1-5 parts of white sugar, and distilled water is added to make the volume to 1000 parts.

[0010] In step 2, the algae cells are cultured in a stepwise manner in a clean bench. The colonies are picked from the algae plate and inoculated into a 10 ml container until the number of algae cells under microscopic examination is ≥ 1.0 × 10 6 / ml, inoculated into a 100ml container and cultured until the number of algal cells under microscopic examination was ≥1.0×10 6 / ml, inoculated into a 1000ml container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into a 5000ml container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into an 18L container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into an 800L container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 After 100 cells / ml, the culture is completed; The ventilation comprises continuous introduction of purified air with a pressure of 0.12 to 0.6 MPa; and intermittent introduction of carbon dioxide with a pressure of 0.12 to 0.6 MPa. The ventilation interval is 3 to 5 hours and the ventilation time is 20 to 50 minutes.

[0011] In step 4, the algae species are compounded and mixed according to the following weight ratios: 20 to 40% of Trisolatum multiflorum 20-60 parts of Micromonas microphylla and 10-30 parts of Chlorella protothecoides; The rotation speed of the compounding and mixing is ≤29r / min.

[0012] The trace element is one of vitamin B1 and vitamin B12 or a combination of the two in any proportion.

[0013] The method for using the microalgae biofertilizer comprises the following steps: pouring the microalgae biofertilizer into water, stirring evenly, and applying to the roots or spraying; The volume ratio of microalgae biofertilizer to water is 1:50-4000; The microalgae biofertilizer comprises one or both of liquid microalgae biofertilizer and powdered microalgae biofertilizer; The stirring speed is 65-150 r / min, and the stirring time is 2-5 min.

[0014] The dosage of microalgae biofertilizer is 500-1200 ml / time / mu.

[0015] Apply microalgae biofertilizer to fertilizers; The fertilizer is one or more of organic fertilizer, bio-organic fertilizer, organic-inorganic fertilizer and water-soluble fertilizer.

[0016] Technical effects of the present invention: Compared with the prior art, the present invention has the following advantages: 1. The combination of Trichoderma variabilis, Chlorella pyrenoidosa and Microcystis pyrenoidosa can better produce nutrients and promote crop growth.

[0017] 2. The present application uses Trisolaria variabilis, Chlorella pyrenoidosa and Microsphaera spp., among which Trisolaria variabilis and Chlorella pyrenoidosa are not protected by a colloid sheath, while Microsphaera spp. is protected by a colloid sheath. Therefore, the present application controls the stirring speed to ≤29r / min during compounding and mixing, in order to prevent the stirring speed from being too fast and destroying Trisolaria variabilis and Chlorella pyrenoidosa; when in use, the stirring speed is 65-150r / min, and the stirring time is 2-5min. Rapid stirring destroys Trisolaria variabilis and Chlorella pyrenoidosa without destroying Microsphaera spp., thereby reducing the absorption of nutrients by algae, and releasing proteins, nucleic acids, vitamins and minerals in the spheroids to provide nutrition for plants. More importantly, after the destruction of Trisolaria variabilis and Chlorella pyrenoidosa, dormant chloroplasts are released and activated to continue photosynthesis, thereby achieving the goal of continuing to produce nutrients and providing nutrition to plants without consuming nutrients. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a diagram showing the microalgae biofertilizer of the present invention. DETAILED DESCRIPTION

[0019] In order to make the objectives, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are clearly and completely described.

[0020] Example 1 The invention discloses a microalgae biofertilizer, which is composed of Micromononuclear Algae, Chlorella Vulgaris and Trisolianthes variegata.

[0021] A method for preparing microalgae biofertilizer comprises the following steps: Step 1: Algal liquid inoculation Inoculate the microalgae into the culture medium and control the temperature, light intensity and pH value; Step 2: Gradual propagation and cultivation The algae species are cultured in stages. When the microalgae cells in the previous stage are ≧1.0*10 6 When the number of cells / ml is reached, the next level of culture is carried out; Step 3: Algal cell screening Use a microscope to test the algae cells to determine whether they are qualified; Unqualified algae cells are inactivated by high temperature treatment and discarded, while qualified algae cells are preserved; sequentially obtain qualified Micromonodactylum cells, qualified Chlorella pyrenoidosa cells, and qualified Trisolianthes variabilis cells; Step 4: Preparation of microalgae biofertilizer Qualified Micromonospora cells, qualified Chlorella pyrenoidosa cells and qualified Trisolianthes variabilis cells are pumped into a mixing tank through a pipeline to compound and mix the algae species to obtain liquid microalgae bio-fertilizer; Freeze-drying the liquid microalgae biofertilizer to obtain powdered microalgae biofertilizer; The freeze-drying conditions are -15-1515°C and the pressure is 200 Pa.

[0022] The temperature was controlled at 25±2℃, the light intensity was 3000±30Lux, and the pH value was 6.8±0.2; The culture medium comprises the following components by weight: 2 parts of sodium bicarbonate, 2 parts of sodium nitrate, 0.8 parts of potassium dihydrogen phosphate, 0.03 parts of calcium disodium ethylenediaminetetraacetate, 0.02 parts of trace elements, and 3 parts of white sugar, and distilled water is added to make the volume to 1000 parts.

[0023] In step 2, the algae cells are cultured in a stepwise manner in a clean bench. The colonies are picked from the algae plate and inoculated into a 10 ml container until the number of algae cells under microscopic examination is ≥ 1.0 × 10 6 / ml, inoculated into a 100ml container and cultured until the number of algal cells under microscopic examination was ≥1.0×10 6 / ml, inoculated into a 1000ml container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into a 5000ml container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into an 18L container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into an 800L container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 After 100 cells / ml, the culture is completed; The ventilation includes continuous introduction of purified air with a pressure of 0.25 MPa; and intermittent introduction of carbon dioxide with a pressure of 0.25 MPa. The ventilation interval is 3.5 hours and the ventilation time is 30 minutes.

[0024] In step 4, the algae species are compounded and mixed according to the following weight ratios: 30 parts of Trisolatum multiflorum, 40 parts of Micromonas microphylla and 20 parts of Chlorella protothecoides; The rotation speed of the compounding and mixing is 15r / min.

[0025] The trace element is vitamin B1.

[0026] The method for using microalgae biofertilizer comprises the following steps: pouring liquid microalgae biofertilizer into water, stirring evenly, Apply with water roots; The volume ratio of microalgae biofertilizer to water is 1:1000; The stirring speed is 65 r / min and the stirring time is 3 min.

[0027] The dosage of liquid microalgae biofertilizer is 1000ml / time / acre.

[0028] Example 2 The invention discloses a microalgae biofertilizer, which is composed of Micromononuclear Algae, Chlorella Vulgaris and Trisolianthes variegata.

[0029] A method for preparing microalgae biofertilizer comprises the following steps: Step 1: Algal liquid inoculation Inoculate the microalgae into the culture medium and control the temperature, light intensity and pH value; Step 2: Gradual propagation and cultivation The algae species are cultured in stages. When the microalgae cells in the previous stage are ≧1.0*10 6 When the number of cells / ml is reached, the next level of culture is carried out; Step 3: Algal cell screening Use a microscope to test the algae cells to determine whether they are qualified; Unqualified algae cells are inactivated by high temperature treatment and discarded, while qualified algae cells are preserved; sequentially obtain qualified Micromonodactylum cells, qualified Chlorella pyrenoidosa cells, and qualified Trisolianthes variabilis cells; Step 4: Preparation of microalgae biofertilizer Qualified Micromonospora cells, qualified Chlorella pyrenoidosa cells and qualified Trisolianthes variabilis cells are pumped into a mixing tank through a pipeline to compound and mix the algae species to obtain liquid microalgae bio-fertilizer; Freeze-drying the liquid microalgae biofertilizer to obtain powdered microalgae biofertilizer; The freeze-drying conditions are -15-1515°C and the pressure is 200 Pa.

[0030] The temperature was controlled at 25±2℃, the light intensity was 3000±30Lux, and the pH value was 6.8±0.2; The culture medium comprises the following components by weight: 2 parts of sodium bicarbonate, 2 parts of sodium nitrate, 0.8 parts of potassium dihydrogen phosphate, 0.03 parts of calcium disodium ethylenediaminetetraacetate, 0.02 parts of trace elements, and 3 parts of white sugar, and distilled water is added to make the volume to 1000 parts.

[0031] In step 2, the algae cells are cultured in a stepwise manner in a clean bench. The colonies are picked from the algae plate and inoculated into a 10 ml container until the number of algae cells under microscopic examination is ≥ 1.0 × 10 6 / ml, inoculated into a 100ml container and cultured until the number of algal cells under microscopic examination was ≥1.0×10 6 / ml, inoculated into a 1000ml container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into a 5000ml container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into an 18L container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into an 800L container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 After 100 cells / ml, the culture is completed; The ventilation includes continuous introduction of purified air with a pressure of 0.25 MPa; and intermittent introduction of carbon dioxide with a pressure of 0.25 MPa. The ventilation interval is 3.5 hours and the ventilation time is 30 minutes.

[0032] In step 4, the algae species are compounded and mixed according to the following weight ratios: 20 parts of Triaspergillus variabilis, 40 parts of Micromonas microphylla, and 30 parts of Chlorella protothecoides; The rotation speed of the compounding and mixing is 15r / min.

[0033] The trace element is vitamin B1.

[0034] The method for using microalgae biofertilizer comprises the following steps: pouring liquid microalgae biofertilizer into water, stirring evenly, Apply with water roots; The volume ratio of microalgae biofertilizer to water is 1:1000; The stirring speed is 65 r / min and the stirring time is 3 min.

[0035] The dosage of liquid microalgae biofertilizer is 1000ml / time / acre.

[0036] Example 3 The invention discloses a microalgae biofertilizer, which is composed of Micromononuclear Algae, Chlorella Vulgaris and Trisolianthes variegata.

[0037] A method for preparing microalgae biofertilizer comprises the following steps: Step 1: Algal liquid inoculation Inoculate the microalgae into the culture medium and control the temperature, light intensity and pH value; Step 2: Gradual propagation and cultivation The algae species are cultured in stages. When the microalgae cells in the previous stage are ≧1.0*10 6 When the number of cells / ml is reached, the next level of culture is carried out; Step 3: Algal cell screening Use a microscope to test the algae cells to determine whether they are qualified; Unqualified algae cells are inactivated by high temperature treatment and discarded, while qualified algae cells are preserved; sequentially obtain qualified Micromonodactylum cells, qualified Chlorella pyrenoidosa cells, and qualified Trisolianthes variabilis cells; Step 4: Preparation of microalgae biofertilizer Qualified Micromonospora cells, qualified Chlorella pyrenoidosa cells and qualified Trisolianthes variabilis cells are pumped into a mixing tank through a pipeline to compound and mix the algae species to obtain liquid microalgae bio-fertilizer; Freeze-drying the liquid microalgae biofertilizer to obtain powdered microalgae biofertilizer; The freeze-drying conditions are -15-1515°C and the pressure is 200 Pa.

[0038] The temperature was controlled at 25±2℃, the light intensity was 3000±30Lux, and the pH value was 6.8±0.2; The culture medium comprises the following components by weight: 2 parts of sodium bicarbonate, 2 parts of sodium nitrate, 0.8 parts of potassium dihydrogen phosphate, 0.03 parts of calcium disodium ethylenediaminetetraacetate, 0.02 parts of trace elements, and 3 parts of white sugar, and distilled water is added to make the volume to 1000 parts.

[0039] In step 2, the algae cells are cultured in a stepwise manner in a clean bench. The colonies are picked from the algae plate and inoculated into a 10 ml container until the number of algae cells under microscopic examination is ≥ 1.0 × 10 6 / ml, inoculated into a 100ml container and cultured until the number of algal cells under microscopic examination was ≥1.0×10 6 / ml, inoculated into a 1000ml container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into a 5000ml container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into an 18L container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into an 800L container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 After 100 cells / ml, the culture is completed; The ventilation includes continuous introduction of purified air with a pressure of 0.25 MPa; and intermittent introduction of carbon dioxide with a pressure of 0.25 MPa. The ventilation interval is 3.5 hours and the ventilation time is 30 minutes.

[0040] In step 4, the algae species are compounded and mixed according to the following weight ratio: 40 parts of Triaspergillus variabilis, 40 parts of Micromonas microphylla, and 20 parts of Chlorella protothecoides; The rotation speed of the compounding and mixing is 15r / min.

[0041] The trace element is vitamin B1.

[0042] The method for using microalgae biofertilizer comprises the following steps: pouring powdered microalgae biofertilizer into water, stirring evenly, and applying it along with the water roots; The volume ratio of powdered microalgae biofertilizer to water is 1:4000; The stirring speed is 65 r / min and the stirring time is 3 min.

[0043] The dosage of powdered microalgae biofertilizer is 250g / time / acre.

[0044] Example 4 The invention discloses a microalgae biofertilizer, which is composed of Micromononuclear Algae, Chlorella Vulgaris and Trisolianthes variegata.

[0045] A method for preparing microalgae biofertilizer comprises the following steps: Step 1: Algal liquid inoculation Inoculate the microalgae into the culture medium and control the temperature, light intensity and pH value; Step 2: Gradual propagation and cultivation The algae species are cultured in stages. When the microalgae cells in the previous stage are ≧1.0*10 6 When the number of cells / ml is reached, the next level of culture is carried out; Step 3: Algal cell screening Use a microscope to test the algae cells to determine whether they are qualified; Unqualified algae cells are inactivated by high temperature treatment and discarded, while qualified algae cells are preserved; sequentially obtain qualified Micromonodactylum cells, qualified Chlorella pyrenoidosa cells, and qualified Trisolianthes variabilis cells; Step 4: Preparation of microalgae biofertilizer The stored qualified Micromonospora cells, qualified Chlorella cells and qualified Trisoliana cells are pumped into a mixing tank through a pipeline to compound and mix the algae species to obtain liquid microalgae bio-fertilizer; Freeze-drying the liquid microalgae biofertilizer to obtain powdered microalgae biofertilizer; The freeze-drying conditions are -15-15°C and the pressure is 300 Pa.

[0046] In step 1, the volume ratio of the algae solution to the culture medium in step 1 is 3:25; The temperature was controlled at 22±2℃, the light intensity was 4000±40Lux, and the pH value was 7.0±0.2; The culture medium comprises the following components by weight: 1 part of sodium bicarbonate, 3 parts of sodium nitrate, 1.2 parts of potassium dihydrogen phosphate, 0.03 parts of calcium disodium ethylenediaminetetraacetate, 0.02 parts of trace elements, and 2 parts of white sugar, and distilled water is added to make the volume to 1000 parts.

[0047] In step 2, the algae cells are cultured in a stepwise manner in a clean bench. The colonies are picked from the algae plate and inoculated into a 10 ml container until the number of algae cells under microscopic examination is ≥ 1.0 × 10 6 / ml, inoculated into a 100ml container and cultured until the number of algal cells under microscopic examination was ≥1.0×10 6 / ml, inoculated into a 1000ml container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into a 5000ml container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into an 18L container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into an 800L container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 After 100 cells / ml, the culture is completed; The ventilation comprises continuous introduction of purified air with a pressure of 0.3 MPa, and intermittent introduction of carbon dioxide with a pressure of 0.32 MPa. The ventilation interval is 4 hours and the ventilation time is 25 minutes.

[0048] In step 4, the algae species are compounded and mixed according to the following weight ratio: 25 parts of Triaspergillus variabilis, 45 parts of Micromonas microphylla, and 20 parts of Chlorella protothecoides; The rotation speed of the compounding and mixing is 10r / min.

[0049] The trace elements are a composition of vitamin B1 and vitamin B12 in a mass ratio of 3:1.

[0050] The method for using microalgae biofertilizer comprises the following steps: pouring liquid microalgae biofertilizer into water, stirring evenly, and applying it along with the water roots; The volume ratio of liquid microalgae biofertilizer to water is 1:1200; The stirring speed is 80 r / min and the stirring time is 3 min.

[0051] The dosage of microalgae biofertilizer is 800ml / time / mu.

[0052] Example 5 The invention discloses a microalgae biofertilizer, which is composed of Micromonospora microphylla, Chlorella pyrenoidosa, Trisolianthes variabilis and Cyclotella meni.

[0053] A method for preparing microalgae biofertilizer comprises the following steps: Step 1: Algal liquid inoculation Inoculate the microalgae into the culture medium and control the temperature, light intensity and pH value; Step 2: Gradual propagation and cultivation The algae species are cultured in stages. When the microalgae cells in the previous stage are ≧1.0*10 6 When the number of cells / ml is reached, the next level of culture is carried out; Step 3: Algal cell screening Use a microscope to test the algae cells to determine whether they are qualified; Unqualified algae cells are inactivated by high temperature treatment and discarded, while qualified algae cells are preserved; sequentially obtaining qualified cells of Micromonospora microphylla, qualified cells of Chlorella pyrenoidosa, qualified cells of Trichoderma variabilis and qualified cells of Cyclotella meni; Step 4: Preparation of microalgae biofertilizer The stored qualified microalgae cells, qualified pyrenoid chlorella cells, qualified triloba cells and meni microcircularia diatoms are pumped into a mixing tank through a pipeline to compound and mix the algae species to obtain liquid microalgae biofertilizer; Freeze-drying the liquid microalgae biofertilizer to obtain powdered microalgae biofertilizer; The freeze-drying conditions are -12-121212°C and the pressure is 300pa.

[0054] In step 1, the volume ratio of the algae solution to the culture medium in step 1 is 4:28; The temperature was controlled at 28±2℃, the light intensity was 3500±35Lux, and the pH value was 6.8±0.2; The culture medium comprises the following components by weight: 4 parts of sodium bicarbonate, 2 parts of sodium nitrate, 1.5 parts of potassium dihydrogen phosphate, 0.05 parts of calcium disodium ethylenediaminetetraacetate, 0.02 parts of trace elements, and 3 parts of white sugar, and distilled water is added to make the volume to 1000 parts.

[0055] In step 2, the algae cells are cultured in a stepwise manner in a clean bench. The colonies are picked from the algae plate and inoculated into a 10 ml container until the number of algae cells under microscopic examination is ≥ 1.0 × 10 6 / ml, inoculated into a 100ml container and cultured until the number of algal cells under microscopic examination was ≥1.0×10 6 / ml, inoculated into a 1000ml container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into a 5000ml container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into an 18L container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into an 800L container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 After 100 cells / ml, the culture is completed; The ventilation comprises continuous introduction of purified air with a pressure of 0.42 MPa, and intermittent introduction of carbon dioxide with a pressure of 0.32 MPa. The ventilation interval is 3 hours and the ventilation time is 40 minutes.

[0056] In step 4, the algae species are compounded and mixed according to the following weight ratios: 30 parts of Trisolatum multiflorum, 50 parts of Micromonas microphylla and 20 parts of Chlorella protothecoides; The rotation speed of the compounding and mixing is ≤29r / min.

[0057] The trace element is vitamin B12.

[0058] The method for using microalgae biofertilizer comprises the following steps: pouring liquid microalgae biofertilizer into water, stirring evenly, and watering the land or roots with the water; The volume ratio of liquid microalgae biofertilizer to water is 1:1100; The stirring speed is 90 r / min and the stirring time is 3 min.

[0059] The dosage of microalgae biofertilizer is 500ml / time / mu.

[0060] The beneficial effects of the present invention are further illustrated below in conjunction with experimental data: Experiment 1 1.1 Experimental location: Laboratory of Shandong Alge Microalgae Biotechnology Co., Ltd.

[0061] 1.2 Experimental test: organic carbon (mg / kg) and total nitrogen (mg / kg).

[0062] 1.3 Test objects: Comparison 1 (except that the Trisoliana cells in Example 1 were replaced by Chlorella pyrenoidosa cells, the other preparation methods were the same), Comparison 2 (except that the Chlorella pyrenoidosa cells in Example 1 were replaced by Trisoliana pyrenoidosa cells, the other preparation methods were the same), and the liquid microalgae bio-fertilizers prepared in Examples 1 and 2.

[0063] 1.4. Experimental method: After the dry soil is passed through a 10-mesh sieve, it is placed in a preheated calciner and heated to 450°C to remove organic matter in the soil; the soil after the organic matter is removed is placed in a container, 100 g is placed in each container, and parallel treatment is performed, with a total of 24 treatments, which are respectively as follows: Comparison 1.1 (except that the Trisoliana cells in Example 1 are replaced with Chlorella pyrenoidosa cells, the other preparation methods are the same) only shake well before absorbing, comparison 1.2 except that the Trisoliana cells in Example 1 are replaced with Chlorella pyrenoidosa cells, the other preparation methods are the same), stirring with a stirrer with a speed of 120 r / min before absorbing; comparison 1.3 (Except that the Trisoliana cells in Example 1 are replaced by Chlorella pyrenoidosa cells, other preparation methods are the same) The mixture is stirred with a stirrer at 200 r / min before absorption; similarly, the following are Comparison 2.1, Comparison 2.2, and Comparison 2.3, Example 1.1, Example 1.2 and Example 1.3, Example 2.1, Example 2.2 and Example 2.3; the potting soil and each treatment correspond randomly, 5 ml of each treatment is absorbed and dropped into the corresponding potting soil, and cultured for 30 days. The culture conditions are a constant temperature of 25° C., a relative humidity of 75%, and a 12 h light / 12 h dark cycle. Organic carbon and total nitrogen are detected after 30 days.

[0064] Note: The difference between Example 1 and Example 2 is that the number of Trichoderma variabilis cells and Chlorella pyrenoidosa cells were exchanged.

[0065] 1.5 Detection method: Total nitrogen is determined by Kjeldahl method; organic carbon is determined by potassium dichromate volumetric method.

[0066] Except for the different treatments, the other implementations of this experiment were the same.

[0067] 2 Results and analysis Organic carbon (mg / kg) and total nitrogen (mg / kg), see Table 1 Table 1

[0068] It can be seen from the data in Table 1 that the organic carbon and total nitrogen contents in the soil treated with Comparative 2 and Example 1, in which the proportion of Trisoliana variabilis cells was greater than the proportion of Chlorella pyrenoidosa cells, were significantly higher than those in Comparative 1 and Example 2, in which the proportion of Trisoliana variabilis cells was greater than the proportion of Chlorella pyrenoidosa cells.

[0069] Promoting plant growth is not only related to total nitrogen and organic matter, but may also be related to biostimulants. A small amount of biostimulants may better promote crop growth and further verification is needed.

[0070] Experiment 2 1.1 Experimental location: 4 mu of vegetable greenhouse in Yingli Town, Shouguang City, Weifang City, the crop planted is Chinese cabbage. The temperature in the experimental greenhouse is 20-28℃, and the humidity is 54-76.

[0071] 1.2 Experimental testing: chlorophyll (SPAD) and yield (kg / mu).

[0072] 1.3 Test objects: blank (watering only), comparison 1 (except that the Trisoliana cells in Example 1 were replaced by Chlorella pyrenoidosa cells, the other preparation methods were the same), comparison 2 (except that the Chlorella pyrenoidosa cells in Example 1 were replaced by Trisoliana pyrenoidosa cells, the other preparation methods were the same), and liquid microalgae bio-fertilizers prepared in Examples 1 and 2.

[0073] 1.4. Experimental method: The 4-acre greenhouse was divided into 24 small areas, each with an area of ​​50 square meters. An isolation zone was set between the small areas. The treatments were respectively as follows: Comparison 1.1 (except that the polymorphic Tri-isocyanate cells in Example 1 were replaced with protein-nucleated Chlorella cells, the other preparation methods were the same) only slightly stirred before use, and the stirring speed was 30 r / min, Comparison 1.2 except that the polymorphic Tri-isocyanate cells in Example 1 were replaced with protein-nucleated Chlorella cells, the other preparation methods were the same), and the stirring speed was 120 r / min before use; Comparison 1.3 (except that the polymorphic Tri-isocyanate cells in Example 1 were replaced with protein-nucleated Chlorella cells, the other preparation methods were the same) before use, the stirring speed was 200 r / min; similarly, comparison 2.1, comparison 2.2, and comparison 2.3, Example 1.1, Example 1.2 and Example 1.3, Example 2.1, Example 2.2 and Example 2.3 The plots were randomly matched to the treatments, and the fertilizer was applied by flushing with water, diluted with water, and the dilution ratio was 1:1000; 200 g of liquid microalgae biofertilizer was applied to each plot.

[0074] On the 12th day after application, chlorophyll was tested using a handheld SPAD meter and the average value was taken. On the 42nd day after sowing, the crops were harvested and the yield was measured and the average value was taken.

[0075] Except for the different treatments, the other implementations of this experiment were the same.

[0076] 2 Results and analysis Chlorophyll (SPAD) and yield (kg / mu), see Table 2 Table 2

[0077] It can be seen from Table 2 that the application of microalgae can increase the chlorophyll content in plants and can increase crop yields. Combined with Table 1, although Comparison 2 has outstanding performance in increasing organic carbon and total nitrogen in Experiment 1, however, in the field test, the effect is not as good as Example 2, which increases organic carbon and total nitrogen less than Comparison 2, and is even worse than Example 1. The possible reasons are: 1. Chlorella pyrenoidosa cells release a certain biostimulant; 2. Trisoliana cells, Chlorella pyrenoidosa cells and Micromonospora microphylla interact with each other and secrete a certain biostimulant; which promotes the growth of crops.

[0078] From the comparison of the data of Example 1 and Example 2, it can be seen that the effect of Example 1 in which the number of Trisoliana variabilis cells is greater than the number of Chlorella pyrenoidosa cells is better than that of Example 2, which is consistent with the data in Table 1.

[0079] It can be seen from the data in Tables 1 and 2 that the stirring speed before use not only affects the output of organic matter, but also affects the growth of crops and the final yield.

[0080] Discussion: The reason for the above results in the present application may be that Trisolaria variabilis and Chlorella pyrenoidosa are not protected by a gelatinous sheath, while Chlorella microcarpa is protected by a gelatinous sheath. Therefore, during compounding and mixing, the present application controls the stirring speed to ≤29r / min, in order to prevent the stirring speed from being too fast and destroying Trisolaria variabilis and Chlorella pyrenoidosa; when in use, the stirring speed is 65-150r / min, and the stirring time is 2-5min. Rapid stirring is used to destroy Trisolaria variabilis and Chlorella pyrenoidosa without destroying Chlorella microcarpa, thereby reducing the absorption of nutrients by algae and releasing proteins, nucleic acids, vitamins and minerals in the spheroids to provide nutrition for plants. What is more important is that after the destruction of Trisolaria variabilis and Chlorella pyrenoidosa, dormant chloroplasts are released and activated to continue photosynthesis, thereby achieving the goal of continuing to produce nutrients and providing nutrition to plants without consuming nutrients.

[0081] The above-mentioned specific implementation modes are only specific cases of the present invention. The patent protection scope of the present invention includes but is not limited to the above-mentioned specific implementation modes. Any appropriate changes or modifications made thereto by any ordinary technician in the technical field in accordance with the claims of the present invention shall fall within the patent protection scope of the present invention.

Claims

1. A microalgae biofertilizer, characterized in that: Including Microcystis, Chlorella and Trichoderma variabilis.

2. The microalgae biofertilizer according to claim 1, characterized in that: The portion of Trisolium variabilis is greater than the portion of Chlorella pyrenoidosa.

3. A method for preparing microalgae biofertilizer according to claim 1 or 2, characterized in that: The following steps are involved: Step 1: Algal liquid inoculation Inoculate the microalgae into the culture medium and control the temperature, light intensity and pH value; Step 2: Gradual propagation and cultivation The algae species are cultured in stages. When the previous level of microalgae cells is ≧1.0*10 6 When the number of cells / ml is reached, the next level of culture is carried out; Step 3: Algal cell screening Use a microscope to test the algae cells to determine whether they are qualified; Unqualified algae cells are inactivated by high temperature treatment and discarded, while qualified algae cells are preserved; Step 4: Preparation of microalgae biofertilizer The preserved qualified algae cells are pumped into a mixing tank through a pipeline for compounding and mixing of algae species to obtain liquid microalgae biofertilizer; Freeze-drying the liquid microalgae biofertilizer to obtain powdered microalgae biofertilizer; The freeze-drying conditions are -20°C to -10°C and the pressure is 100 to 500 Pa; Microalgae biofertilizer includes liquid microalgae biofertilizer and powder microalgae biofertilizer.

4. The method for preparing microalgae biofertilizer according to claim 3, characterized in that: In step 1, the volume ratio of the algae solution to the culture medium in step 1 is 1-5:20-30; The temperature was controlled at 20-30°C, the light intensity was 2000-5000 Lux, and the pH value was 6.5-7.5; The culture medium comprises the following components by weight: 1-5 parts of sodium bicarbonate, 1-5 parts of sodium nitrate, 0.8-2 parts of potassium dihydrogen phosphate, 0.01-0.05 parts of calcium disodium ethylenediaminetetraacetate, 0.01-0.05 parts of trace elements, 1-5 parts of white sugar, and distilled water is added to make the volume to 1000 parts; The trace element is one of vitamin B1 and vitamin B12 or a combination of the two in any proportion.

5. The method for preparing microalgae biofertilizer according to claim 3, characterized in that: In step 2, the algae cells are cultured in a stepwise manner in a clean bench. The colonies are picked from the algae plate and inoculated into a 10 ml container until the number of algae cells under microscopic examination is ≥ 1.0 × 10 6 / ml, inoculated into a 100ml container and cultured until the number of algal cells under microscopic examination was ≥1.0×10 6 / ml, inoculated into a 1000ml container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into a 5000ml container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into an 18L container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 / ml, inoculated into an 800L container for aeration culture until the number of algal cells under microscopic examination is ≥1.0×10 6 After 100 cells / ml, the culture is completed; The ventilation comprises continuous introduction of purified air with a pressure of 0.12 to 0.6 MPa; and intermittent introduction of carbon dioxide with a pressure of 0.12 to 0.6 MPa. The ventilation interval is 3 to 5 hours and the ventilation time is 20 to 50 minutes.

6. The method for preparing microalgae biofertilizer according to claim 3, characterized in that: In step 4, press the following The algae species are compounded and mixed according to the proportion of the components: including 20-40 parts of Trichoderma variabilis, 20-60 parts of Micromonospora microphylla, and 10-30 parts of Chlorella protothecoides; The rotation speed of the compounding and mixing is ≤29r / min.

7. The method for using the microalgae biofertilizer according to any one of claims 1 to 6, comprising the following steps: Microalgae Pour the biofertilizer into the water, stir evenly, and apply it to the roots or by spraying; The volume ratio of microalgae biofertilizer to water is 1:50-4000; The microalgae biofertilizer includes one or both of liquid microalgae biofertilizer and powdered microalgae biofertilizer; The stirring speed is 65-150 r / min, and the stirring time is 2-5 min.

8. The method for using the microalgae biofertilizer according to claim 7, wherein the amount of the microalgae biofertilizer used is 500-1200 ml / time / acre.

9. The method for using the microalgae biofertilizer according to claim 7 or 8, characterized in that: Apply microalgae biofertilizer to fertilizers; The fertilizer is one or more of organic fertilizer, bio-organic fertilizer, organic-inorganic fertilizer and water-soluble fertilizer.

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