A plant growth regulator for tomatoes and preparation method thereof

Abstract

The present invention discloses a plant growth regulator for tomatoes and a preparation method thereof, and belongs to the technical field of plant growth regulators. The plant growth regulator of the present invention comprises the following raw materials in parts by weight: 30-50 parts of algae extract, 20-40 parts of bacterial culture, 3-6 parts of chitosan, 0.1-0.5 parts of sodium alginate, and 100-150 parts of auxiliary materials. The plant growth regulator of the present invention, when used in small amounts, can greatly increase crop yield and quality, while reducing the probability of occurrence of tomato gray mold, solving the problems of slow growth of tomatoes, low yield and high incidence of diseases in some tomato planting areas, and the ingredients are safe and environmentally friendly, non-toxic to crops, and have a positive effect on reducing environmental pollution and pesticide residues.

Description

Technical Field

[0001] The invention belongs to the technical field of plant growth regulators, and particularly relates to a plant growth regulator for tomatoes and a preparation method thereof. Background Art

[0002] Tomato is one of the most important vegetables. Its fruit is rich in nutrients, including carbohydrates, organic acids, and vitamins needed by the human body. It has health benefits such as anti-cancer and blood pressure reduction. However, in the early spring, when the temperature is low and the light is weak, tomatoes often suffer from problems such as flower and fruit drop. The application of plant growth regulators can effectively solve the problems of flower and fruit drop, slow plant growth, low fruit setting rate, and reduced yield.

[0003] Plant growth regulators refer to beneficial substances that exist naturally in plants and have a significant effect on plant growth and development. They are also called natural plant hormones or endogenous plant hormones, which can affect and control plant growth. They maintain plant growth from multiple links such as cell growth and division, germination, flowering, and fruiting, and can control the entire process. Since the current large-scale cultivation of tomatoes is mainly facility cultivation, the effect of pollination and fruit setting under natural conditions is not ideal, and the yield is low. Therefore, plant growth regulators are widely used in the tomato planting process to improve pollination and fruit setting rates, preserve flowers and fruits, and increase yields.

[0004] For example, Chinese patent application CN202210483877.5 discloses a growth regulator composition for tomatoes, which includes the following components by weight: 5-10 parts of chlormequat, 10-15 parts of triacontanol, 5-8 parts of paclobutrazol, and 10-12 parts of jasmonate compounds. Compared with the prior art, the present invention is extensive: a variety of growth regulators are applied to the field of tomatoes, which can effectively play the role of different regulators, improve the growth conditions and planting environment of plants, and effectively improve their yield and quality;

[0005] However, some of the current existing technologies mostly use common growth regulators and other chemical components in combination, with a single function. And repeated and long-term use of such regulators often causes plant poisoning, which has a great impact on the growth, yield and quality of tomatoes. The symptoms of plant growth regulator poisoning in tomatoes are often manifested as curled and deformed leaves. When the poisoning symptoms are severe, they appear as fern leaves, twisted leaves, or even linear. The appearance of tomato plant growth regulator poisoning symptoms is similar to viral diseases and is often misjudged as a virus by vegetable farmers. Therefore, the safety of this type of regulator is difficult to guarantee and is not suitable for long-term use.

[0006] Therefore, developing a green and safe plant growth regulator while optimizing its single growth-promoting effect is a technical problem that needs to be solved urgently. Summary of the invention

[0007] In view of the problems existing in the prior art, the present invention provides a safe and environmentally friendly plant growth regulator, which uses algae extracts and microorganisms as main functional substances. On the one hand, it effectively promotes the growth of tomatoes and improves the quality; on the other hand, it effectively inhibits the main pathogenic fungus of tomato gray mold - Botrytis cinerea, and improves crop resistance.

[0008] In order to achieve the above technical objectives, the technical solution adopted by the present invention is:

[0009] A plant growth regulator for tomatoes comprises the following raw materials in parts by weight: 30-50 parts of algae extract, 20-40 parts of bacterial culture, 3-6 parts of chitosan, 0.1-0.5 parts of sodium alginate and 100-150 parts of auxiliary materials.

[0010] Furthermore, the preparation method of the algae extract is: accurately weigh 30g of Ascophyllum nodosum powder and 30g of Chlorella vulgaris powder, put them into a container, add 300mL of a 3% phosphoric acid solution, extract at 80-90°C for 3-5h, then add 7-10g of potassium hydroxide, stir for 30-40min, and centrifuge at 8000-10000r / min in a high-speed centrifuge for 5-10min, and the supernatant is the algae extract.

[0011] Furthermore, the preparation method of the bacterial culture is:

[0012] Strain activation: The Epicoccum nigrum and Bacillus cereus stored in the refrigerator were transferred into LB culture medium and placed at 25-30°C for activation culture for 24 hours to obtain activated strains of Epicoccum nigrum and Bacillus cereus;

[0013] Seed liquid culture: take a loop of activated E. melanogaster and Bacillus cereus, inoculate them into 250mL conical flasks filled with seed culture medium, fill them with 20% liquid, place them in a shaking incubator at 25-30°C and 200r / min for 12-13h to obtain seed liquid of the two bacteria, mix the seed liquids of the two bacteria in a volume ratio of 1:1 to obtain a mixed seed liquid;

[0014] Fermentation culture: Inoculate the mixed seed liquid into a 1L conical flask containing fermentation medium at a 1% inoculation rate, fill it with 50% liquid, and culture it in a shaking incubator at 25-30℃ and 200r / min. Ferment and culture until the effective viable count is 2.4-3.2×10 9 cfu / ml, and adjust the pH value to 6.5-7.5 to obtain bacterial culture.

[0015] Furthermore, the strain number of the black coccus in step (1) is CGMCC No. 3.13856, purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration; the strain number of the Bacillus cereus is CGMCC No. 1.15914, purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration.

[0016] The selected Echinococcus melanococcus and Bacillus cereus can be purchased through the open search of the strain catalog of the General Microbiological Center of the China Culture Collection Administration, without the need for repeated biological deposits.

[0017] Furthermore, the composition of the seed culture medium in step (2) is: 10.0 g·L tryptone –1 , yeast extract powder 5.0 g•L –1 , NaCl10.0g•L –1 , pH 7.2-7.4, high temperature sterilization at 121℃ for 20min.

[0018] Furthermore, the composition of the fermentation medium in step (3) is: glucose 20.0 g·L –1 , peptone 10.0 g•L –1 , KH2PO41.0g•L –1 ,MgSO41.0g•L –1 ,MnSO4·H2O 0.2 g·L –1 , sterilize at 115℃ for 20min.

[0019] Furthermore, the auxiliary material is obtained by mixing sodium lignin sulfonate, sodium dodecyl sulfate, urea, starch and water in a mass ratio of 0.1:0.3:3:5:20.

[0020] A method for preparing a plant growth regulator for tomatoes, comprising the following preparation steps:

[0021] (1) preparing algae extract;

[0022] (2) preparing bacterial cultures;

[0023] (3) mixing sodium lignin sulfonate, sodium dodecyl sulfate, urea, starch, and water in the auxiliary materials in a mass ratio of 0.1:0.3:3:5:20, and adding chitosan to obtain a premix;

[0024] (4) Add the algae extract, bacterial culture and sodium alginate into the premix and mix them evenly.

[0025] The method for using the plant growth regulator of the present invention is as follows: starting from tomato planting, 20 L / mu is poured on the roots of the plants.

[0026] Beneficial Effects

[0027] The present invention excludes the use of traditional compound plant growth regulators, and extracts and uses the active ingredients of Ascophyllum nodosum and Chlorella vulgaris as one of the functional ingredients. In the Ascophyllum nodosum extract, it contains a variety of functional organic molecules that dissolve nutrients and promote growth, such as betaine and its various derivatives, which can promote the early growth and vigorous growth of plants, and also contains rich stress-resistant growth-promoting substances, such as alginic acid. These substances work together to improve the resistance of plants to environmental stress, promote root development, nutrient absorption and water utilization. And Chlorella vulgaris extract, it is rich in nutrients such as protein, amino acids, nucleotides, polysaccharides and vitamins. These rich nutrients can enhance the growth and development of plants, and at the same time it has good antioxidant properties, can effectively remove free radicals in plants, and reduce the damage of oxidative stress to plant cells, which helps to protect plant cells from pathogens and enhance their stress resistance. The two algae extracts are used together to show obvious disease-resistant growth-promoting effects.

[0028] At the same time, the present invention selects black Echinococcus and Bacillus cereus to form functional microorganisms. First, the two bacteria have good antagonistic effects on the main pathogenic bacteria of tomatoes-Botrytis cinerea. Secondly, the seed liquid of the two is mixed in equal proportions and then fermented. During the fermentation process, they promote each other and produce a variety of antifungal compounds. These compounds can significantly inhibit the growth of pathogens and synergistically inhibit bacteria. At the same time, these functional growth-promoting agents also have the functions of solubilizing phosphorus, secreting indoleacetic acid (IAA) and protease, etc. These functions play an important role in the growth and development of plants.

[0029] The present invention also adds sodium lignin sulfonate, sodium dodecyl sulfate, urea, starch and other substances as auxiliary materials, which can provide certain nutrients for the soil on the one hand, and on the other hand, sodium lignin sulfonate, starch and other substances provide energy for microorganisms, significantly improve the release rate and survival time of the bacteria, thereby enhancing the overall growth promotion and control effect.

[0030] The plant growth regulator of the present invention can greatly improve crop yield and quality when used in a small amount, while reducing the probability of occurrence of tomato gray mold, solving the problems of slow growth, low yield and high incidence of diseases in some tomato planting areas. The ingredients are safe and environmentally friendly, non-toxic to crops, and have a positive effect on reducing environmental pollution and pesticide residues. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 The antibacterial effect diagram of E. melanogaster and Bacillus cereus strains;

[0032] Figure 2 This is a diagram showing the effects of phosphate solubilization and protein decomposition of E. melanogaster;

[0033] Figure 3 This is a diagram showing the effects of Bacillus cereus on solubilizing phosphate and decomposing protein;

[0034] Figure 4 This is a diagram showing the antibacterial effects of different fermentation bacterial broths. DETAILED DESCRIPTION

[0035] The technical solution of the present invention is further described below in conjunction with specific embodiments, but is not limited thereto.

[0036] Example 1

[0037] A plant growth regulator for tomatoes comprises the following raw materials in parts by weight: 50 parts of algae extract, 40 parts of bacterial culture, 6 parts of chitosan, 0.1 parts of sodium alginate and 100 parts of auxiliary materials.

[0038] The preparation method of the algae extract is as follows: accurately weigh 30g of Ascophyllum nodosum powder and 30g of Chlorella vulgaris powder, put them into a container, add 300mL of a phosphoric acid solution with a mass concentration of 3%, extract at 80-90°C for 5h, then add 10g of potassium hydroxide, stir for 40min, centrifuge at 10000r / min for 10min in a high-speed centrifuge, and the supernatant is the algae extract.

[0039] The preparation method of the bacterial culture is:

[0040] Strain activation: The black Echinococcus and Bacillus cereus stored in the refrigerator were transferred into LB culture medium and placed at 25-30°C for activation culture for 24 hours to obtain activated strains of black Echinococcus and Bacillus cereus;

[0041] Seed liquid culture: take a loop of activated E. melanogaster and Bacillus cereus, inoculate them into 250mL conical flasks filled with seed culture medium, fill them with 20% liquid, place them in a shaking incubator at 25-30°C and 200r / min for 12h to obtain seed liquid of the two bacteria, mix the seed liquids of the two bacteria in a volume ratio of 1:1 to obtain a mixed seed liquid;

[0042] Fermentation culture: Inoculate the mixed seed liquid into a 1L conical flask containing fermentation medium at a 1% inoculation rate, fill it with 50% liquid, and culture it in a shaking incubator at 25-30℃ and 200r / min. Ferment and culture until the effective viable count is 2.4-3.2×10 9 cfu / ml, and adjust the pH value to 6.5-7.5 to obtain bacterial culture.

[0043] The strain number of the black coccus in step (1) is CGMCC No. 3.13856, which was purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration; the strain number of the Bacillus cereus is CGMCC No. 1.15914, which was purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration.

[0044] The selected Echinococcus melanococcus and Bacillus cereus can be purchased through the open search of the strain catalog of the General Microbiological Center of the China Culture Collection Administration, without the need for repeated biological deposits.

[0045] Step (2) The composition of the seed culture medium is: 10.0 g·L tryptone –1 , yeast extract powder 5.0 g•L –1 , NaCl10.0g•L –1 , pH 7.2-7.4, high temperature sterilization at 121℃ for 20min.

[0046] Step (3) The composition of the fermentation medium is: glucose 20.0 g·L –1 , peptone 10.0 g•L –1 , KH2PO41.0g•L –1 ,MgSO41.0g•L –1 ,MnSO4·H2O 0.2 g·L –1 , sterilize at 115℃ for 20min.

[0047] The auxiliary materials are sodium lignin sulfonate, sodium dodecyl sulfate, urea, starch and water mixed in a mass ratio of 0.1:0.3:3:5:20.

[0048] A method for preparing a plant growth regulator for tomatoes, comprising the following preparation steps:

[0049] (1) preparing algae extract;

[0050] (2) preparing bacterial cultures;

[0051] (3) mixing sodium lignin sulfonate, sodium dodecyl sulfate, urea, starch, and water in the auxiliary materials in a mass ratio of 0.1:0.3:3:5:20, and adding chitosan to obtain a premix;

[0052] (4) Add the algae extract, bacterial culture and sodium alginate into the premix and mix them evenly.

[0053] Example 2

[0054] A plant growth regulator for tomatoes comprises the following raw materials in parts by weight: 40 parts of algae extract, 30 parts of bacterial culture, 3 parts of chitosan, 0.5 parts of sodium alginate and 120 parts of auxiliary materials.

[0055] The preparation method of the algae extract is as follows: accurately weigh 30g of Ascophyllum nodosum powder and 30g of Chlorella vulgaris powder, put them into a container, add 300mL of a phosphoric acid solution with a mass concentration of 3%, extract at 80-90°C for 3h, then add 7g of potassium hydroxide, stir for 30min, centrifuge at 8000r / min for 5min in a high-speed centrifuge, and the supernatant is the algae extract.

[0056] The preparation method of the bacterial culture is:

[0057] Strain activation: The black Echinococcus and Bacillus cereus stored in the refrigerator were transferred into LB culture medium and placed at 25-30°C for activation culture for 24 hours to obtain activated strains of black Echinococcus and Bacillus cereus;

[0058] Seed liquid culture: take a loop of activated E. melanogaster and Bacillus cereus, inoculate them into 250mL conical flasks filled with seed culture medium, fill them with 20% liquid, place them in a shaking incubator at 25-30°C and 200r / min for 13h to obtain seed liquid of the two bacteria, mix the seed liquids of the two bacteria in a volume ratio of 1:1 to obtain a mixed seed liquid;

[0059] Fermentation culture: Inoculate the mixed seed liquid into a 1L conical flask containing fermentation medium at a 1% inoculation rate, fill it with 50% liquid, and culture it in a shaking incubator at 25-30℃ and 200r / min. Ferment and culture until the effective viable count is 2.4-3.2×10 9 cfu / ml, and adjust the pH value to 6.5-7.5 to obtain bacterial culture.

[0060] The strain number of the black coccus in step (1) is CGMCC No. 3.13856, which was purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration; the strain number of the Bacillus cereus is CGMCC No. 1.15914, which was purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration.

[0061] The selected Echinococcus melanococcus and Bacillus cereus can be purchased through the open search of the strain catalog of the General Microbiological Center of the China Culture Collection Administration, without the need for repeated biological deposits.

[0062] Step (2) The composition of the seed culture medium is: 10.0 g·L tryptone –1 , yeast extract powder 5.0 g•L –1 , NaCl10.0g•L–1 , pH 7.2-7.4, high temperature sterilization at 121℃ for 20min.

[0063] Step (3) The composition of the fermentation medium is: glucose 20.0 g·L –1 , peptone 10.0 g•L –1 , KH2PO41.0g•L –1 ,MgSO41.0g•L –1 ,MnSO4·H2O 0.2 g·L –1 , sterilize at 115℃ for 20min.

[0064] The auxiliary materials are sodium lignin sulfonate, sodium dodecyl sulfate, urea, starch and water mixed in a mass ratio of 0.1:0.3:3:5:20.

[0065] A method for preparing a plant growth regulator for tomatoes, comprising the following preparation steps:

[0066] (1) preparing algae extract;

[0067] (2) preparing bacterial cultures;

[0068] (3) mixing sodium lignin sulfonate, sodium dodecyl sulfate, urea, starch, and water in the auxiliary materials in a mass ratio of 0.1:0.3:3:5:20, and adding chitosan to obtain a premix;

[0069] (4) Add the algae extract, bacterial culture and sodium alginate into the premix and mix them evenly.

[0070] Example 3

[0071] A plant growth regulator for tomatoes comprises the following raw materials in parts by weight: 45 parts of algae extract, 35 parts of bacterial culture, 4 parts of chitosan, 0.3 parts of sodium alginate and 150 parts of auxiliary materials.

[0072] The preparation method of the algae extract is as follows: accurately weigh 30g of Ascophyllum nodosum powder and 30g of Chlorella vulgaris powder, put them into a container, add 300mL of a phosphoric acid solution with a mass concentration of 3%, extract at 80-90°C for 4h, then add 8g of potassium hydroxide, stir for 35min, centrifuge at 9000r / min for 8min in a high-speed centrifuge, and the supernatant is the algae extract.

[0073] The preparation method of the bacterial culture is:

[0074] Strain activation: The black Echinococcus and Bacillus cereus stored in the refrigerator were transferred into LB culture medium and placed at 25-30°C for activation culture for 24 hours to obtain activated strains of black Echinococcus and Bacillus cereus;

[0075] Seed liquid culture: take a loop of activated E. melanogaster and Bacillus cereus, inoculate them into 250mL conical flasks filled with seed culture medium, fill them with 20% liquid, place them in a shaking incubator at 25-30°C and 200r / min for 12-13h to obtain seed liquid of the two bacteria, mix the seed liquids of the two bacteria in a volume ratio of 1:1 to obtain a mixed seed liquid;

[0076] Fermentation culture: Inoculate the mixed seed liquid into a 1L conical flask containing fermentation medium at a 1% inoculation rate, fill it with 50% liquid, and culture it in a shaking incubator at 25-30℃ and 200r / min. Ferment and culture until the effective viable count is 2.4-3.2×10 9 cfu / ml, and adjust the pH value to 6.5-7.5 to obtain bacterial culture.

[0077] The strain number of the black coccus in step (1) is CGMCC No. 3.13856, which was purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration; the strain number of the Bacillus cereus is CGMCC No. 1.15914, which was purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration.

[0078] The selected Echinococcus melanococcus and Bacillus cereus can be purchased through the open search of the strain catalog of the General Microbiological Center of the China Culture Collection Administration, without the need for repeated biological deposits.

[0079] Step (2) The composition of the seed culture medium is: 10.0 g·L tryptone –1 , yeast extract powder 5.0 g•L –1 , NaCl10.0g•L –1 , pH 7.2-7.4, high temperature sterilization at 121℃ for 20min.

[0080] Step (3) The composition of the fermentation medium is: glucose 20.0 g·L –1 , peptone 10.0 g•L –1 , KH2PO41.0g•L –1 ,MgSO41.0g•L –1 ,MnSO4·H2O 0.2 g·L –1 , sterilize at 115℃ for 20min.

[0081] The auxiliary materials are sodium lignin sulfonate, sodium dodecyl sulfate, urea, starch and water mixed in a mass ratio of 0.1:0.3:3:5:20.

[0082] A method for preparing a plant growth regulator for tomatoes, comprising the following preparation steps:

[0083] (1) preparing algae extract;

[0084] (2) preparing bacterial cultures;

[0085] (3) mixing sodium lignin sulfonate, sodium dodecyl sulfate, urea, starch, and water in the auxiliary materials in a mass ratio of 0.1:0.3:3:5:20, and adding chitosan to obtain a premix;

[0086] (4) Add the algae extract, bacterial culture and sodium alginate into the premix and mix them evenly.

[0087] Example 4

[0088] A plant growth regulator for tomatoes comprises the following raw materials in parts by weight: 30 parts of algae extract, 20 parts of bacterial culture, 3 parts of chitosan, 0.5 parts of sodium alginate and 100 parts of auxiliary materials.

[0089] The preparation method of the algae extract is as follows: accurately weigh 30g of Ascophyllum nodosum powder and 30g of Chlorella vulgaris powder, put them into a container, add 300mL of a phosphoric acid solution with a mass concentration of 3%, extract at 80-90°C for 3h, then add 7g of potassium hydroxide, stir for 30min, centrifuge at 8000r / min for 5min in a high-speed centrifuge, and the supernatant is the algae extract.

[0090] The preparation method of the bacterial culture is:

[0091] Strain activation: The black Echinococcus and Bacillus cereus stored in the refrigerator were transferred into LB culture medium and placed at 25-30°C for activation culture for 24 hours to obtain activated strains of black Echinococcus and Bacillus cereus;

[0092] Seed liquid culture: take a loop of activated E. melanogaster and Bacillus cereus, inoculate them into 250mL conical flasks filled with seed culture medium, fill them with 20% liquid, place them in a shaking incubator at 25-30°C and 200r / min for 12-13h to obtain seed liquid of the two bacteria, mix the seed liquids of the two bacteria in a volume ratio of 1:1 to obtain a mixed seed liquid;

[0093] Fermentation culture: Inoculate the mixed seed liquid into a 1L conical flask containing fermentation medium at a 1% inoculation rate, fill it with 50% liquid, and culture it in a shaking incubator at 25-30℃ and 200r / min. Ferment and culture until the effective viable count is 2.4-3.2×10 9 cfu / ml, and adjust the pH value to 6.5-7.5 to obtain bacterial culture.

[0094] The strain number of the black coccus in step (1) is CGMCC No. 3.13856, which was purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration; the strain number of the Bacillus cereus is CGMCC No. 1.15914, which was purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration.

[0095] The selected Echinococcus melanococcus and Bacillus cereus can be purchased through the open search of the strain catalog of the General Microbiological Center of the China Culture Collection Administration, without the need for repeated biological deposits.

[0096] Step (2) The composition of the seed culture medium is: 10.0 g·L tryptone –1 , yeast extract powder 5.0 g•L –1 , NaCl10.0g•L –1 , pH 7.2-7.4, high temperature sterilization at 121℃ for 20min.

[0097] Step (3) The composition of the fermentation medium is: glucose 20.0 g·L –1 , peptone 10.0 g•L –1 , KH2PO41.0g•L –1 ,MgSO41.0g•L –1 ,MnSO4·H2O 0.2 g·L –1 , sterilize at 115℃ for 20min.

[0098] The auxiliary materials are sodium lignin sulfonate, sodium dodecyl sulfate, urea, starch and water mixed in a mass ratio of 0.1:0.3:3:5:20.

[0099] A method for preparing a plant growth regulator for tomatoes, comprising the following preparation steps:

[0100] (1) preparing algae extract;

[0101] (2) preparing bacterial cultures;

[0102] (3) mixing sodium lignin sulfonate, sodium dodecyl sulfate, urea, starch, and water in the auxiliary materials in a mass ratio of 0.1:0.3:3:5:20, and adding chitosan to obtain a premix;

[0103] (4) Add the algae extract, bacterial culture and sodium alginate into the premix and mix them evenly.

[0104] Comparative Example 1

[0105] A plant growth regulator for tomatoes comprises the following raw materials in parts by weight: 30 parts of algae extract, 20 parts of bacterial culture, 3 parts of chitosan, 0.5 parts of sodium alginate and 100 parts of auxiliary materials.

[0106] The preparation method of the algae extract is as follows: accurately weigh 60g of chlorella powder, put it into a container, add 300mL of a phosphoric acid solution with a mass concentration of 3%, extract at 80-90°C for 3h, then add 7g of potassium hydroxide, stir for 30min, centrifuge at 8000r / min for 5min in a high-speed centrifuge, and the supernatant is the algae extract.

[0107] In this comparative example, except that only Chlorella powder was used in the algae extract, the other raw materials and preparation methods were the same as those in Example 4.

[0108] Comparative Example 2

[0109] A plant growth regulator for tomatoes comprises the following raw materials in parts by weight: 30 parts of algae extract, 20 parts of bacterial culture, 3 parts of chitosan, 0.5 parts of sodium alginate and 100 parts of auxiliary materials.

[0110] The preparation method of the algae extract is as follows: accurately weigh 60g of Ascophyllum nodosum powder, put it into a container, add 300mL of a phosphoric acid solution with a mass concentration of 3%, extract at 80-90°C for 3h, then add 7g of potassium hydroxide, stir for 30min, centrifuge at 8000r / min for 5min in a high-speed centrifuge, and the supernatant is the algae extract.

[0111] In this comparative example, except that only Ascophyllum nodosum powder was used in the algae extract, the other raw materials and preparation methods were the same as those in Example 4.

[0112] Comparative Example 3

[0113] A plant growth regulator for tomatoes comprises the following raw materials in parts by weight: 30 parts of algae extract, 20 parts of bacterial culture, 3 parts of chitosan, 0.5 parts of sodium alginate and 100 parts of auxiliary materials.

[0114] The preparation method of the algae extract is as follows: accurately weigh 30g of Ascophyllum nodosum powder and 30g of Chlorella vulgaris powder, put them into a container, add 300mL of a phosphoric acid solution with a mass concentration of 3%, extract at 80-90°C for 3h, then add 7g of potassium hydroxide, stir for 30min, centrifuge at 8000r / min for 5min in a high-speed centrifuge, and the supernatant is the algae extract.

[0115] The preparation method of the bacterial culture is:

[0116] Strain activation: transfer the black Echinococcus stored in the refrigerator into LB culture medium and place it at 25-30°C for activation culture for 24 hours to obtain the black Echinococcus activated strain;

[0117] Seed liquid culture: Take a loop of activated black Escherichia cocci strain and inoculate it into a 250mL conical flask filled with seed culture medium, fill it with 20% liquid, and culture it in a shaking incubator at 25-30℃ and 200r / min for 12-13h to obtain seed liquid;

[0118] Fermentation culture: Inoculate the seed liquid at a 1% inoculum into a 1L conical flask filled with fermentation medium, fill it with 50% liquid, and culture it in a shaking incubator at 25-30℃ and 200r / min. Ferment and culture until the effective viable count is 2.4-3.2×10 9 cfu / ml, and adjust the pH value to 6.5-7.5 to obtain bacterial culture.

[0119] In this comparative example, except that only E. nigrosinus was used for fermentation in the bacterial culture, the other raw materials and preparation method were the same as those in Example 4.

[0120] Comparative Example 4

[0121] A plant growth regulator for tomatoes comprises the following raw materials in parts by weight: 30 parts of algae extract, 20 parts of bacterial culture, 3 parts of chitosan, 0.5 parts of sodium alginate and 100 parts of auxiliary materials.

[0122] The preparation method of the algae extract is as follows: accurately weigh 30g of Ascophyllum nodosum powder and 30g of Chlorella vulgaris powder, put them into a container, add 300mL of a phosphoric acid solution with a mass concentration of 3%, extract at 80-90°C for 3h, then add 7g of potassium hydroxide, stir for 30min, centrifuge at 8000r / min for 5min in a high-speed centrifuge, and the supernatant is the algae extract.

[0123] The preparation method of the bacterial culture is:

[0124] Strain activation: The Bacillus cereus stored in the refrigerator was transferred into LB medium and placed at 25-30°C for activation culture for 24 hours to obtain an activated strain of Bacillus cereus;

[0125] Seed liquid culture: Take a loop of activated Bacillus cereus strain and inoculate it into a 250mL conical flask filled with seed culture medium, fill it with 20% liquid, and culture it in a shaking incubator at 25-30℃ and 200r / min for 12-13h to obtain seed liquid;

[0126] Fermentation culture: Inoculate the seed liquid at a 1% inoculum into a 1L conical flask filled with fermentation medium, fill it with 50% liquid, and culture it in a shaking incubator at 25-30℃ and 200r / min. Ferment and culture until the effective viable count is 2.4-3.2×10 9 cfu / ml, and adjust the pH value to 6.5-7.5 to obtain bacterial culture.

[0127] In this comparative example, except that only Bacillus cereus was used for fermentation in the bacterial culture, the other raw materials and preparation method were the same as those in Example 4.

[0128] Comparative Example 5

[0129] A plant growth regulator for tomatoes comprises the following raw materials in parts by weight: 30 parts of algae extract, 20 parts of bacterial culture, 3 parts of chitosan, 0.5 parts of sodium alginate and 100 parts of auxiliary materials.

[0130] The preparation method of the algae extract is as follows: accurately weigh 30g of Ascophyllum nodosum powder and 30g of Chlorella vulgaris powder, put them into a container, add 300mL of a phosphoric acid solution with a mass concentration of 3%, extract at 80-90°C for 3h, then add 7g of potassium hydroxide, stir for 30min, centrifuge at 8000r / min for 5min in a high-speed centrifuge, and the supernatant is the algae extract.

[0131] The preparation method of the bacterial culture is:

[0132] Strain activation: The black Echinococcus and Bacillus cereus stored in the refrigerator were transferred into LB culture medium and placed at 25-30°C for activation culture for 24 hours to obtain activated strains of black Echinococcus and Bacillus cereus;

[0133] Seed liquid culture: take a loop of activated E. melanogaster and Bacillus cereus, inoculate them into 250mL conical flasks filled with seed culture medium, fill them with 20% liquid, place them in a shaking incubator at 25-30°C and 200r / min for 12-13h to obtain seed liquid of the two bacteria, mix the seed liquids of the two bacteria in a volume ratio of 2:1 to obtain a mixed seed liquid;

[0134] Fermentation culture: Inoculate the mixed seed liquid into a 1L conical flask containing fermentation medium at a 1% inoculation rate, fill it with 50% liquid, and culture it in a shaking incubator at 25-30℃ and 200r / min. Ferment and culture until the effective viable count is 2.4-3.2×10 9 cfu / ml, and adjust the pH value to 6.5-7.5 to obtain bacterial culture.

[0135] In this comparative example, except for changing the dosage ratio of E. nigrosinus and Bacillus cereus in the bacterial culture, the other raw materials and preparation method are the same as those in Example 4.

[0136] Comparative Example 6

[0137] A plant growth regulator for tomatoes comprises the following raw materials in parts by weight: 30 parts of algae extract, 20 parts of bacterial culture, 3 parts of chitosan, 0.5 parts of sodium alginate and 100 parts of auxiliary materials.

[0138] The preparation method of the algae extract is as follows: accurately weigh 30g of Ascophyllum nodosum powder and 30g of Chlorella vulgaris powder, put them into a container, add 300mL of a phosphoric acid solution with a mass concentration of 3%, extract at 80-90°C for 3h, then add 7g of potassium hydroxide, stir for 30min, centrifuge at 8000r / min for 5min in a high-speed centrifuge, and the supernatant is the algae extract.

[0139] The preparation method of the bacterial culture is:

[0140] Strain activation: The black Echinococcus and Bacillus cereus stored in the refrigerator were transferred into LB culture medium and placed at 25-30°C for activation culture for 24 hours to obtain activated strains of black Echinococcus and Bacillus cereus;

[0141] Seed liquid culture: take a loop of activated E. melanogaster and Bacillus cereus, inoculate them into 250mL conical flasks filled with seed culture medium, fill them with 20% liquid, place them in a shaking incubator at 25-30°C and 200r / min for 12-13h to obtain seed liquid of the two bacteria, mix the seed liquids of the two bacteria in a volume ratio of 1:2 to obtain a mixed seed liquid;

[0142] Fermentation culture: Inoculate the mixed seed liquid into a 1L conical flask containing fermentation medium at a 1% inoculation rate, fill it with 50% liquid, and culture it in a shaking incubator at 25-30℃ and 200r / min. Ferment and culture until the effective viable count is 2.4-3.2×10 9 cfu / ml, and adjust the pH value to 6.5-7.5 to obtain bacterial culture.

[0143] In this comparative example, except for changing the dosage ratio of E. nigrosinus and Bacillus cereus in the bacterial culture, the other raw materials and preparation method are the same as those in Example 4.

[0144] Performance Testing

[0145] Verification of antibacterial effect of strains

[0146] The plate confrontation method was used to determine the antibacterial activity of the strain, using the pathogen of tomato gray mold, Botrytis cinerea, as the target. A 5mm diameter pathogenic fungus cake was picked and inoculated in the center of the PDA medium. A dipped inoculation loop was used to inoculate black E. cocci and Bacillus cereus at a distance of 2cm from the center of the plate. No bacteria were inoculated as a control. After culturing in a constant temperature incubator at 25-30℃ for 3 days, the diameter of the pathogenic fungus colony was measured and the antibacterial rate was calculated. Each treatment and test was repeated 3 times, and the values ​​were averaged.

[0147] Antibacterial rate%=(C d -T d )×100 / (Cd -0.5cm); C d (cm): diameter of the control pathogenic fungal colony; T d (cm): diameter of the treated pathogenic fungal colony.

[0148] Table 1 Antibacterial effect of strains

[0149]

[0150] From the table data and Figure 1 It can be seen that the E. nigrosinus and Bacillus cereus screened by the present invention both have good antagonistic effects on the pathogen of tomato gray mold.

[0151] Determination of antibacterial activity of different fermentation broths

[0152] The bacterial cultures obtained in Example 4 and Comparative Examples 3 to 6 were used as samples to verify the antibacterial activity of fermentation broths composed of different strains. The experimental method is as follows:

[0153] After disinfection, place the prepared PDA culture medium, culture dish and Oxford cup on a sterile workbench. After 15 minutes of ultraviolet irradiation, put 20 ml of the culture medium directly into the culture dish and continue to irradiate with ultraviolet light for 15 minutes. After the culture medium is completely solidified, place 1 ml of the prepared gray mold suspension on the culture medium, and then use a spreader to evenly apply it on the culture medium. Then place it vertically in the Oxford cup, contacting the culture medium until there is no gap. Pipette 100 μl of the bacterial culture to be tested and add it to the Oxford cup. After culturing in a 25-30℃ incubator for 10 days, observe the antibacterial effect. Effects such as Figure 4 It can be seen that after the mixture of E. melanogaster and Bacillus cereus in equal proportions and fermentation, the two produce a synergistic effect and have a high antibacterial effect.

[0154] Determination of related metabolites of antagonistic strains

[0155] Taking the seed solution obtained in Example 4 as an example, the phosphate-dissolving inorganic phosphorus culture medium was poured onto a plate, a hole was punched in the center with a 9 mm puncher, and 50 μL of seed solution was added into the hole with a pipette. After culturing in a constant temperature incubator at 25-30°C for 3 days, the formation of a phosphate-dissolving ring was observed.

[0156] Extracellular protease production assay

[0157] Pour skim milk powder culture medium into plates, punch a hole in the center with a 9 mm borer, add 50 μL of seed solution into the hole with a pipette, and culture in a constant temperature incubator at 25-30°C for 3 days to observe whether a transparent circle is produced.

[0158] from Figure 2-Figure 3 It can be seen that both strains have good abilities of solubilizing phosphate and producing protease.

[0159] Determination of indoleacetic acid production ability:

[0160] The IAA biosynthesis capacity of the strain was determined by the Salkowski method. First, the strain was inoculated in a solution containing 200 mg·L - 1 In the NA liquid medium containing L-tryptophan, the temperature was kept at 28℃ and 200r·min. - 1 The samples were cultured on a shaker for 3 days. A low-temperature centrifuge was used at 4°C and 10,000 r / min - 1 After centrifugation for 10 min, the supernatant was removed and added with an equal volume of Salkowski colorimetric solution. After standing for 30 min in the dark, the absorbance was measured at a wavelength of 530 nm using an ultraviolet spectrophotometer to determine the IAA synthesis capacity of the strain.

[0161] Table 2 Determination of growth-promoting ability

[0162]

[0163] Pot experiment

[0164] Tomato variety tested: “Qingnong 866”.

[0165] The test soil was collected from a greenhouse in the experimental base of the Linyi University Scientific Research Institute in Linyi City, Shandong Province, where tomatoes had been grown continuously for 10 years. Gray mold disease often occurs here. The five-point sampling method was used for sampling, and the samples were sieved to remove stones and other debris before use.

[0166] Test samples: plant regulators obtained from Examples 1 to 4 and Comparative Examples 1 to 6, and a blank control was also set up.

[0167] Experimental methods:

[0168] The experimental flower pots were plastic flower pots with an upper inner diameter of 23 cm and a height of 21 cm, purchased from Shandong Jinan Xiangrui Gardening Supplies Co., Ltd.

[0169] The experiment set up 11 treatments, each treatment was repeated five times, and the results were averaged:

[0170] Treatment 1: 2 mL of the regulator of Example 1 per pot + planting normal tomato seedlings;

[0171] Treatment 2: 2 mL of the regulator of Example 2 per pot + planting normal tomato seedlings;

[0172] Treatment 3: 2 mL of the regulator of Example 3 per pot + planting normal tomato seedlings;

[0173] Treatment 4: 2 mL of the regulator of Example 4 per pot + planting normal tomato seedlings;

[0174] Treatment 5: 2 mL of the regulator of comparative example 1 per pot + planting normal tomato seedlings;

[0175] Treatment 6: 2 mL of the regulator of comparative example 2 per pot + planting normal tomato seedlings;

[0176] Treatment 7: 2 mL of the regulator of comparative example 3 per pot + planting normal tomato seedlings;

[0177] Treatment 8: 2 mL of the regulator of comparative example 4 per pot + planting normal tomato seedlings;

[0178] Treatment 9: 2 mL of the regulator of comparative example 5 per pot + planting normal tomato seedlings;

[0179] Treatment 10: 2 mL of the regulator of comparative example 6 per pot + planting normal tomato seedlings;

[0180] Treatment 11: 2 mL of clean water per pot + planting normal tomato seedlings.

[0181] After planting, water each pot with 100 mL of water, and then water each pot with 100 mL of water every 2 days. Conventional cultivation management was adopted, which was consistent among treatments. After planting, the growth and disease status of the plants were observed regularly.

[0182] When the tomato seedlings grow to the seven-leaf stage, the disease level of the plants is recorded according to the grading standards for tomato gray mold, and the disease index and disease prevention effect are calculated.

[0183] Disease prevention effect (%) = (disease index of control area - disease index of treated area) / disease index of control area × 100.

[0184] During the tomato fruiting period, 5 fruits (per plant) were randomly picked and brought back to the laboratory for tomato quality index determination, including Vc, protein, soluble sugar, etc. The superoxide dismutase (SOD) activity was determined by the nitroblue tetrazolium method, the peroxidase (POD) activity was determined by the guaiacol method, and the catalase (CAT) activity was determined by the ultraviolet absorption method.

[0185] Table 3 Potted plant experiment results (1)

[0186]

[0187] Table 4 Potted plant experiment results (2)

[0188]

[0189] From the data in the table, we can see that the quality and disease resistance of the tomatoes obtained in the examples of the present invention are significantly better than those of the blank control. This shows that the plant growth regulator of the present invention can effectively prevent and control tomato gray mold, improve crop resistance, and at the same time increase the content of tomato VC, protein and soluble sugar, thereby improving crop quality. However, in Comparative Examples 1-2, which have changed the composition of algae extracts, and Comparative Examples 3-4, which have changed the composition of microorganisms, the synergistic balance between the effective ingredients and the microbial agents is broken, and the macroscopic manifestation is that the regulatory effect on plants is weakened, and the crop resistance and quality are reduced.

[0190] It should be noted that the above embodiments are only partial embodiments of the preferred methods of implementing the present invention, rather than all embodiments. Obviously, based on the above embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

Claims

1. A plant growth regulator for tomatoes, characterized in that: The invention comprises the following raw materials in parts by weight: 30-50 parts of algae extract, 20-40 parts of bacterial culture, 3-6 parts of chitosan, 0.1-0.5 parts of sodium alginate, and 100-150 parts of auxiliary materials; the preparation method of the algae extract is as follows: accurately weigh 30g of Ascophyllum nodosum powder and 30g of Chlorella vulgaris powder, put them into a container, add 300mL of 3% phosphoric acid solution, extract at 80-90℃ for 3-5h, then add 7-10g of potassium hydroxide, stir for 30-40min, centrifuge at 8000-10000r / min for 5-10min in a high-speed centrifuge, and the supernatant is the algae extract; the preparation method of the bacterial culture is as follows: (1) Strain activation: Activate the black Echinococcus ( Epicoccum nigrum ) and Bacillus cereus ( Bacillus cereus ) were respectively transferred into LB medium and placed at 25-30°C for activation culture for 24 hours to obtain activated strains of E. melanogaster and Bacillus cereus; (2) Seed liquid culture: Take a loop of activated E. melanogaster and Bacillus cereus, respectively, and inoculate into 250 mL conical flasks filled with seed culture medium, fill with 20% liquid, place in a shaking incubator at 25-30°C and 200 rpm for 12-13 h to obtain seed liquid of the two bacteria, and mix the seed liquids of the two bacteria in a volume ratio of 1:1 to obtain a mixed seed liquid; (3) Fermentation culture: The mixed seed liquid was inoculated at 1% into a 1L conical flask containing fermentation medium, and the liquid volume was 50%. The flask was placed in a shaking incubator at 25-30°C and 200 r / min for fermentation until the effective viable bacterial count reached 2.4-3.2×10 9 cfu / ml, and the pH was adjusted to 6.5-7.5 to obtain bacterial culture; The black Echinococcus ( Epicoccum nigrum ) strain number is CGMCC No.3.13856, purchased from the General Microbiology Center of China National Microbiological Culture Collection Administration; the Bacillus cereus ( Bacillus cereus ) strain number is CGMCC No.1.15914, which was purchased from the General Microbiology Center of China Culture Collection Administration.

2. The plant growth regulator for tomatoes according to claim 1, characterized in that: The auxiliary materials are sodium lignin sulfonate, sodium dodecyl sulfate, urea, starch and water mixed in a mass ratio of 0.1:0.3:3:5:

20.

3. A method for preparing the plant growth regulator for tomatoes according to any one of claims 1 to 2, characterized in that: The method comprises the following preparation steps: (1) preparing algae extract; (2) preparing bacterial cultures; (3) mixing sodium lignin sulfonate, sodium dodecyl sulfate, urea, starch, and water in the auxiliary materials in a mass ratio of 0.1:0.3:3:5:20, and adding chitosan to obtain a premix; (4) Add the algae extract, bacterial culture and sodium alginate into the premix and mix them evenly.

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