A method for producing a composite bacterial plant growth regulator

Abstract

A method for manufacturing a composite bacterial plant growth regulator belongs to the technical field of microbial fertilizers. With components such as Laxogenin, Laxogenin C and Laxogenin derivatives, glucose, starch, EDTA-iron, EDTA-zinc, DA series, organosilicon, lithium oxide, etc., after fusion dispersion under ultrapure water liquid phase, it is warmed to 30-85°C, slowly added with hydrogen peroxide for hydroxylation reaction, and after keeping the temperature for 30-120min, it is cooled to room temperature, and then a bacterial agent is added, such as: yeast water agent, and the composite bacterial plant growth regulator is fully stirred and dispersed to obtain the composite bacterial plant growth regulator. The composite bacterial plant growth regulator produced by the present invention can be used for foliar spraying, drip irrigation or hydroponics, can effectively improve the plant growth state, can also improve the soil environment, while improving plant resistance, improving crop traits and up-regulating various metabolic functions, efficiently promoting root and leaf to promote chlorophyll synthesis, effectively improving tillering number and disease and insect resistance, therefore, compared with other chemical regulation and regulation substances, it also has pleiotropic function and wide-spectrum applicability.

Description

Technical Field

[0001] The present invention relates to a composite bacterial plant growth regulator and a manufacturing method thereof, belonging to the technical field of microbial fertilizers, and specifically to a composite plant growth regulator containing multiple components. The composite bacterial plant growth regulator of the present invention can be widely used in field crops, such as rice, wheat, rapeseed, soybean, and corn, and can effectively and significantly improve per-acre yield and resistance to stress, diseases, and pests; it can also be used in the fields of garden flower cultivation, hydroponics, fruit, tea, and greenhouse drip irrigation, and can effectively promote rooting, flowering, fruit preservation, and tillering. Background Art

[0002] After decades of using chemical fertilizers and pesticides, modern agriculture has severely damaged the living environment of plants. Water bodies have also been polluted to varying degrees, causing many originally fertile fields to gradually become less productive and the quality of food to decline year by year. It also leads to crop inversion and an increase in the frequency of insect pests throughout the life cycle. The current mainstream agricultural technology for increasing production and income is basically to improve the resistance / tolerance of plants and promote crop production by using various macro / trace elements and plant hormones / growth regulators. The effect of improving various agronomic traits in this way is not stable and is often accompanied by certain harmful effects.

[0003] The source of the component LA and its derivatives of the present invention is to extract the wild plant of Smilax china, which grows in low mountain shrubs or valley river banks, is widely distributed in northern regions, and has abundant resources. The natural product Laxogenin and its derivatives can effectively promote the growth and development of plants and the formation of morphology, enhance various metabolic capabilities, increase yield and quality, and can improve plant resistance / stress tolerance, improve plant yield and quality, enhance plant salt resistance, insect resistance and drought resistance and other physiological effects, and show the significant biological activity of brassinosteroids (BRs). Laxogenin and its derivatives are recognized as pollution-free green and environmentally friendly plant growth agents. Laxogenin and its derivatives themselves are derived from medicinal plant resources, belong to plant endogenous natural metabolites, can be absorbed and metabolized by plants themselves, and therefore will not cause adverse effects on the ecological environment, nor will they cause human health and safety issues. It is a non-toxic and pollution-free plant growth regulation technology that meets the needs of green and environmentally friendly agriculture.

[0004] The invented compound beneficial bacteria will effectively improve the living environment of crops, efficiently promote the photosynthesis of plants, accelerate the solubility of phosphorus, potassium and nitrogen, resist diseases and insect pests, and have extremely strong resistance / stress tolerance.

[0005] The DA series is hormone-free and can increase the activity of enzymes in plants, increase photosynthesis, increase the content of chlorophyll and nucleic acids, and promote plant cell division and growth. EDAT iron, zinc sulfate, lithium, etc. are one of the main elements for plant growth and development, and are also one of the necessary elements for fragrance and fruit setting. Trace lithium is beneficial to delay the aging process of cells and enhance cell vitality. At the same time, lithium can increase the activity of enzymes in plants, increase photosynthesis, increase the content of chlorophyll and nucleic acids, and promote plant cell division and growth. Summary of the invention

[0006] The composite bacterial plant growth regulator of the present invention makes up for the deficiencies of traditional crop production regulators by using Laxogenin and its derivatives, beneficial bacteria and a combination of specific stress-resistant elements, and effectively reduces nitrogen fertilizers, phosphorus fertilizers such as urea, ammonium bicarbonate, ammonium phosphate, compound fertilizers and other chemical fertilizers and pesticides. It improves the activity of the plant's own cells, improves the growth of microbial flora, effectively improves the living environment of crops, efficiently promotes photosynthesis of plants, accelerates phosphorus, potassium and nitrogen dissolution, resists diseases and insect pests, and has extremely strong resistance / stress tolerance; greatly improves the activity of enzymes in plants, enhances photosynthesis, increases the content of chlorophyll and nucleic acids, promotes the cell division ability and growth activity of plants, and thus improves the quality and yield of crop production; and achieves the purpose of increasing production and income and green ecology.

[0007] The object of the present invention is achieved by: a method for preparing a composite bacterial plant growth regulator,

[0008] (1) dispersing and dissolving Laxogenin (LA), LG-C or their derivatives in ultrapure water in advance;

[0009] The structural formula of Laxogenin (LA).

[0010] (2) Add glucose, starch, EDTA-iron, DA series, silicone, zinc sulfate, lithium hydroxide, etc. in sequence, raise the temperature to 30-85°C, and stir thoroughly to disperse and dissolve;

[0011] (3) maintaining the solution temperature of step (2) at 30-85° C., slowly adding hydrogen peroxide to carry out hydroxylation reaction of LA for 30-120 min (the other components act as auxiliary agents or catalysts for the hydroxylation reaction and will also become beneficial elements for crops);

[0012] (4) In step (3), the material is cooled to below 45° C., and then a bacterial agent is added, such as yeast, photosynthetic bacteria, Streptomyces jingyangensis (5406), Streptomyces rubrotinctum, Bacillus megaterium, Bacillus jelly, Bacillus subtilis, Bacillus licheniformis, etc. The mass fraction of the bacterial agent is 10-15%.

[0013] The raw materials are calculated by weight: 0.001-0.01 parts of Laxogenin LA, 10-15 parts of glucose, 5-10 parts of starch, 10-30 parts of EDTA iron, 0.01-0.1 parts of DA, 20-30 parts of zinc sulfate, 0.01-0.05 parts of lithium source and 0.5-1 parts of organosilicon.

[0014] The organic silicon in step (2) includes polyether silicone oil or ethoxy-modified trisiloxane.

[0015] The lithium source in step (2) includes one or more of lithium hydroxide, lithium sulfide, lithium chloride, lithium nitrate and lithium phosphate.

[0016] The DA series described in step (2) includes DA-6, DA-7, or DA-8.

[0017] The mass fraction of the hydrogen peroxide in step (3) is 40-50%; the added amount of the hydrogen peroxide with a mass fraction of 40-50% is 500-2000 times the mass of Laxogenin LA.

[0018] The hydroxylation reaction conditions in the step (3) are 45-85° C. and 30-120 min.

[0019] The composite bacterial plant growth regulator for gardening and agricultural crops obtained by the method provided by the present invention has a light yellow transparent solution in appearance and emits a light aromatic odor; the main elements thereof are detected by ICP: K≤50ppm, heavy metals such as Cu, Pb, Cd, Cr≤10ppm; Ca, Mg≤500ppm, Zn, B are respectively ≥1500ppm.

[0020] The composite bacterial plant growth regulator produced by the present invention has easy-to-control quality, low cost, simple process, safety, greenness and environmental protection, and can effectively improve the ecological environment for plant survival. The obtained product is highly concentrated, and only 100 ml per standard acre can fully achieve the purpose of improving quality and increasing yield, and has excellent cost performance and reliability.

[0021] The composite bacterial plant growth regulator of the present invention makes up for the deficiencies of traditional crop production regulators by using Laxogenin and its derivatives, beneficial bacteria and a combination of specific stress-resistant elements, effectively reduces chemical fertilizers and pesticides, increases the activity of the plant's own cells, improves the growth of microbial flora, and thus improves the quality and yield of crop production. DETAILED DESCRIPTION

[0022] The implementation process of the present invention is as follows: first, Laxogenin (LA), Laxogenin C (LG-C) and its derivatives are pre-dispersed and dissolved in ultrapure water, and after the temperature is raised to 30-85°C, glucose, starch, EDTA-iron, EDTA-zinc, DA series, organosilicon, lithium borate (or boric acid, lithium hydroxide) and the like are added in sequence, and the mixture is fully stirred, dispersed and dissolved. The temperature is maintained at 30-85°C, and 50% hydrogen peroxide is slowly and quantitatively added to carry out hydroxyl-reaction reaction. After the hydrogen peroxide is added, the reaction is continued for 30-120 minutes, the material is cooled to below 45°C, and the bacterial agent is added. After the material is fully dispersed, it is filled and stored in a light-proof and cool environment for use.

[0023] The bacterial agent of the present invention includes yeast, photosynthetic bacteria, Jingyang Streptomyces 5406, red and yellow Streptomyces, Bacillus megaterium, jelly-like Bacillus, Bacillus subtilis, and Bacillus licheniformis. Among them, yeast was purchased from Angel Yeast Co., Ltd. Photosynthetic bacteria were purchased from Qingdao Xuneng Bioengineering Co., Ltd. Jingyang Streptomyces 5406 was purchased from Shanghai Fusheng Biotechnology Development Co., Ltd. Red and yellow Streptomyces was purchased from Henan Haomeite Biotechnology Co., Ltd. Bacillus megaterium, jelly-like Bacillus, Bacillus subtilis, and Bacillus licheniformis were purchased from Jinan Jinyuyuan Biotechnology Co., Ltd.

[0024] Example 1

[0025] A method for preparing a composite bacterial plant growth regulator comprises the following steps:

[0026] 1. Weigh 0.01g LA and dissolve it in 10ml ethanol, then transfer it to a 2500ml beaker filled with 1000ml ultrapure water and stir thoroughly to decompose evenly.

[0027] 2. Heat the previous solution to 45°C and maintain the temperature, then add 5g of food-grade glucose, 5g of water-soluble starch, 10g of industrial-grade 99% food-grade EDTA iron, 10g of 98% zinc sulfate, 0.1g of DA-6 (amino fresh fat), 10ml of 10% ethoxy-modified trisiloxane silicone solvent, and 0.05g of lithium hydroxide in sequence; control the stirring speed to 45-63r / min and fully disperse and dissolve for 15 minutes.

[0028] 3. After the dissolution temperature in the previous step is raised to 65°C, slowly add 7.5g of industrial grade 50% hydrogen peroxide at a rate of 0.5-0.8g / min.

[0029] 4. After the addition of hydrogen peroxide, the temperature was maintained to continue the reaction for 45 minutes, and then the heating was stopped and the mixture was naturally cooled to below 45°C. This step completed the hydroxylation reaction to obtain a compound containing 5A-hydroxy Laxogenin. The compound was subjected to three-stage extraction and distillation to obtain a colorless refined liquid, which was further concentrated to obtain fine white particles. The sample was detected by liquid spectroscopy analysis and was determined to be 5A-hydroxy Laxogenin, and its structural formula is:

[0030]

[0031] 5. Keep stirring at 45-63r / min, add 0.2g of photosynthetic bacteria, 0.2g of Jingyang Streptomyces (5406), 0.1g of red and yellow Streptomyces, 0.1g of Bacillus megaterium, 0.05g of gelatinous Bacillus, 0.05g of Bacillus subtilis, and 0.1g of Bacillus licheniformis, and continue stirring for 30 minutes; after the synthesized solution is cooled to room temperature, it is a composite bacterial plant growth regulator, which is filled in a brown PP bottle and sealed for storage.

[0032] The dosage per standard acre varies slightly depending on the application method. The recommended dosage is 10-30ml / standard acre, and the preferred dosage is 20ml / standard acre. For foliar spraying with flying control, dilute it at 1:1, for manual foliar spraying, dilute it at 1:50, and for hydroponics, root dipping or drip irrigation, dilute it at 1:100.

[0033] The composite bacterial plant growth regulator obtained in Example 1 was used to explore the effect of promoting rice growth and increasing rice yield by dipping the roots of rice seedlings, providing a basis for large-scale promotion and application. From early June to late September, the soil is flooded paddy soil with medium fertility, sufficient sunlight and sufficient water. Demonstration variety: Hongyou No. 7.

[0034] Demonstration Materials and Methods:

[0035] The demonstration set up 4 plots, 3 treatments and 2 controls. The dosage of compound bacterial plant growth regulator in the treatment area was 10ml, 20ml and 30ml respectively, corresponding to treatment 1, treatment 2 and treatment 3. Control 1 was no application, and control 2 was LA without hydroxylation treatment, and the dosage was 20ml. Each plot area was 100m 2 Earth ridges are set up to separate the plots, and protection rows are set up outside the experimental area.

[0036] 3. Demonstration method

[0037] Before transplanting and planting (June 7), the compound bacteria plant growth regulator treatment area measured the corresponding compound bacteria plant growth regulator dosage tube and diluted it at 1:50, poured it into a plastic pot, and all the seedlings in the plot were dipped in the pot until the roots were completely dipped, and then transplanted. The first and second production surveys were conducted on June 27 and July 17: 3 bags were taken from each plot, and the average maximum value of the main stem and the number of stem tillers in each bag were measured for the aboveground part; the average maximum value of the main root was measured for the underground part, and the average root weight was taken after washing and air-drying. (See Table 1 and Table 2). The seeds were tested on September 24 and the theoretical measurements were determined (see Table 3).

[0038] As can be seen from Table 2, after the seedlings were treated with the (composite bacterial plant growth regulator) of Example 1 for 40 days, the plant height, stem tiller number, root length and root weight of rice were effectively improved. The main stem height of rice in treatment 1 was increased by 4.3% on average compared with the control area, and the stem tiller number was increased by 23.2% compared with the control area; the main root length was increased by 16.0% compared with the control, and the root weight was increased by 106.9% compared with the control.

[0039] As can be seen from Table 3, the application of the (composite bacterial plant growth regulator) in Example 1 to rice can effectively increase the yield of rice. Treatment 1 increased the yield by an average of 40.6% compared with the control.

[0040] The results of this demonstration show that the application of Chuzhiyi (a composite bacterial plant growth regulator) to rice can effectively promote rice growth (stronger in the middle stage than in the early stage), which can greatly increase rice yield and can be promoted and applied on a large scale in production.

[0041] Table 1. Survey form of rice growth in treated rice (June 27)

[0042] Rice Process 1 Process 2 Process 3 Comparison 1 Comparison 2 Main stem height 50.6 57.8 46.3 55.8 52.8 Number of tillers 54 45 55 52 49 Taproot length 20.2 19.2 19.5 22.5 21.3 Root weight 22.1 11.6 22.8 26.2 23.4

[0043] Table 2 Survey form for rice growth in treated areas (July 17)

[0044] Rice Process 1 Process 2 Process 3 Comparison 1 Comparison 2 Main stem height 72.3 72.3 71.7 69.3 70.6 Number of tillers 69 59 56 56 55 Taproot length 24.7 20.7 20.0 21.3 22.6 Root weight 48.2 37.0 33.7 23.3 31.8

[0045] Table 3 Survey of rice yield in treated areas (September 24)

[0046] Rice Total number of ears (10,000 plants / mu) Fruiting rate (%) Number of grains per ear Yield per mu (kg) Process 1 16.3 76.24 185.1 675.7 Process 2 14.2 76.17 166.3 529.4 Process 3 14.4 76.35 179.3 578.0 Comparison 1 13.3 74.35 162.4 480.4 Comparison 2 14.0 74.8 165.2 507.3

[0047] The finished product obtained by the above synthesis was applied to the roots of hybrid rice in the Jiangling experimental field of the Agricultural Technology Extension Center of Jingzhou City, Hubei Province, according to the above usage and dosage, and 25 days later, the main stem height of rice increased by 4.3% on average, the number of tillers increased by 23.2% on average, the main root length increased by 16% on average, the root weight increased by 106% on average, and the yield increased by 10.6% on average compared with the control.

[0048] The ICP test results of the composite bacterial plant growth regulator obtained in Example 1 are shown in Table 4:

[0049] Table 4

[0050]

[0051] Example 2

[0052] 1. Weigh 0.005g LA and dissolve it in 10ml ethanol, then transfer it to a 2500ml beaker filled with 1000ml ultrapure water and stir thoroughly to decompose evenly;

[0053] 2. Heat the solution in the previous step to 45°C and maintain the temperature, then add 5g of food-grade glucose, 5g of water-soluble starch, 10g of industrial-grade 99% food-grade EDTA iron, 25g of 98% zinc sulfate, 10ml of 10% organosilicon solvent, 25g of boric acid, and 0.025g of lithium hydroxide in sequence; control the stirring speed to 45-63r / min and fully disperse and dissolve for 15 minutes;

[0054] 3. After the dissolution temperature in the previous step is raised to 65°C, slowly add 5g of industrial grade 50% hydrogen peroxide at a rate of 0.5-0.8g / min;

[0055] 4. After the addition of hydrogen peroxide, the temperature was maintained for 45 minutes, and then the heating was stopped and the mixture was naturally cooled to below 45°C. This step completed the hydroxylation reaction and obtained 5A-hydroxy Laxogenin.

[0056] 5. Keep stirring at 45-63r / min, add 0.15g photosynthetic bacteria, 0.2g Jingyang Streptomyces (5406), 0.2g red and yellow Streptomyces, and 0.2g Bacillus megaterium;

[0057] 6. Continue stirring for 30 minutes;

[0058] 7. After the synthetic solution is cooled to room temperature, it is the composite bacterial plant growth regulator. Fill the solution into a brown PP bottle and seal it for later use. For foliar spraying of this product, dilute it at 1:1, 20ml / 667m 2 Usage: dilute 1:50 for artificial foliage spraying, 1:100 for hydroponics, root dipping or drip irrigation.

[0059] The finished product obtained by the above synthesis was used at the Sinochem Shandong Linyi Crop Center during the wheat germination period at the above dosage to simulate flight control for 28 days, and organic and inorganic fertilizers (15-40S) were used as controls.

[0060] The average main stem height of wheat increased by 3.7% compared with the control, the average number of tillers increased by 17.4% compared with the control, the average main root length increased by 7.8% compared with the control, the root weight increased by 85% compared with the control, and the average yield increased by 8.7% compared with the control. For specific effects, please refer to the data in Tables 5, 6, 7 and 8 below.

[0061] Table 5 Treatment table of wheat sand culture test

[0062]

[0063] Table 6 Effects of different treatments on wheat plant height

[0064] deal with 14 days 19 days 28 days T1 29.11±1.45 29.89±2.39 33.08±2.26 T2 21.69±2.01 28.32±2.69 34.17±1.52 T3 26.30±1.98 29.16±1.53 33.56±3.09 T4 26.94±2.36 27.39±2.14 31.96±3.85 T5 23.26±1.31 27.85±2.71 34.26±1.75 T6 23.49±4.28 29.17±1.84 34.00±2.42 T7 24.47±3.13 25.37±3.18 28.44±2.30 T8 24.66±2.26 28.32±1.46 32.78±2.12 T9 22.96±3.96 27.60±2.52 33.84±3.16 T10 26.71±3.38 31.48±2.59 37.09±2.24 CK 23.82±2.24 28.58±2.35 36.83±1.54

[0065] Table 7 Effects of different treatments on wheat root length

[0066] deal with 14 days 19 days 28 days T1 12.87±2.06 15.17±2.91 17.31±2.06 T2 12.48±2.92 13.41±3.88 17.67±1.85 T3 15.61±1.65: 17.84±3.62 17.77±2.6 T4 12.68±2.22 15.70±2.56 17.15±2.28 T5 11.61±3.74. 11.12±3.00 14.04±1.71 T6 14.32±2.78 17.71±2.06 18.98±3.22 T7 12.93±4.73 14.58±4.46 14.93±3.21 T8 14.60±3.01 16.69±3.97 15.18±1.53 T9 14.49±2.00 15.57±2.54 13.11±2.25 T10 15.95±2.22 16.73±1.89 20.23±3.15 CK 11.91±2.15 14.04±3.13 14.81±1.89

[0067] Table 8 Effects of different treatments on wheat biomass

[0068]

[0069] The ICP test results of the obtained composite bacterial plant growth regulator are shown in the following table:

[0070]

[0071]

[0072] The composite bacterial plant growth regulator of the present invention makes up for the deficiencies of traditional crop regulators by using Laxogenin and its derivatives, beneficial bacteria and a combination of specific stress-resistant elements, overcomes the use of various macro- / trace elements and plant hormones / growth regulators in the prior art, effectively reduces chemical fertilizers and pesticides, improves the activity of plant cells, improves the growth of microbial flora, effectively improves the living environment of crops, efficiently promotes photosynthesis of plants, accelerates phosphorus, potassium and nitrogen dissolution, resists diseases and insect pests, and has extremely strong resistance / stress tolerance; greatly improves the activity of enzymes in plants, enhances photosynthesis, increases the content of chlorophyll and nucleic acid, promotes the cell division ability and growth activity of plants, and thus improves the quality and yield of crop production; and achieves the purpose of increasing production and income and green ecology.

[0073] Under the enlightenment of the present invention, other various forms of products can be obtained. Therefore, the above specific implementation methods should not be understood as limiting the scope of protection of the present invention. The scope of protection of the present invention should be based on the definition in the claims, and the description can be used to interpret the claims. The above is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention can have various changes and variations. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection of the present invention.

Claims

1. A method for producing a composite bacterial plant growth regulator, characterized in that: The steps include: (1) Weigh 0.01 g of LA and dissolve it in 10 ml of ethanol. Transfer it to a 2500 ml beaker containing 1000 ml of ultrapure water and stir thoroughly to decompose it evenly. (2) Heat the solution in the previous step to 45°C and maintain the temperature, then add 5g of food-grade glucose, 5g of water-soluble starch, 10g of industrial-grade 99% food-grade EDTA iron, 10g of 98% zinc sulfate, 0.1g of DA-6 amine fresh fat, 10ml of 10% ethoxy-modified trisiloxane silicone solvent, and 0.05g of lithium hydroxide in sequence; control the stirring speed to 45-63r / min and fully disperse and dissolve for 15 minutes; (3) After the dissolution temperature in the previous step is raised to 65°C, 7.5 g of industrial grade 50% hydrogen peroxide is slowly added at a rate of 0.5-0.8 g / min; (4) After the addition of hydrogen peroxide, the temperature was maintained to continue the reaction for 45 minutes, and then the heating was stopped and the mixture was naturally cooled to below 45°C. This step completed the hydroxylation reaction and obtained a compound containing 5A-hydroxy Laxogenin. The compound was subjected to three-stage extraction and distillation to obtain a colorless refined liquid, which was further concentrated to obtain fine white particles. The sample was detected by liquid chromatography and was determined to be 5A-hydroxy Laxogenin, with the structural formula being: ； (5) Maintain stirring at 45-63 r / min, add 0.2 g of photosynthetic bacteria, 0.2 g of Jingyang Streptomyces 5406, 0.1 g of erythrocytes, 0.1 g of Bacillus megaterium, 0.05 g of Bacillus gelatinous, 0.05 g of Bacillus subtilis, and 0.1 g of Bacillus licheniformis, and continue stirring for 30 minutes; after the synthesized solution is cooled to room temperature, it is a composite bacterial plant growth regulator, which is filled into a brown PP bottle and sealed for storage.

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