A seed coating agent, a preparation method thereof, and application thereof

Abstract

The application discloses a seed coating agent, a preparation method and application thereof, and belongs to the field of agricultural technology.The seed coating agent comprises the following components by weight: 1-5 parts of a thickening agent, 2-10 parts of a film forming agent, 0.2-1 part of a dyeing agent, 0.01-0.05 part of a bittering agent, 5-25 parts of a preparation agent and 100-1500 parts of a nitrogen-fixing bacteria liquid.The seed coating agent is mixed with seeds and stirred uniformly, and then is dried in the shade and is ready for sowing.The components are mixed and stirred uniformly during preparation.When the seed coating agent is applied, the activity of the nitrogen-fixing bacteria contained in the seed coating agent can be preserved for more than 20 days;when the seed coating agent is applied, the seed coating agent can promote growth and yield, and the application of the seed coating agent can significantly promote the growth and yield of crops in low-fertility soil, but has no significant effect on crops in fertile soil, and is more suitable for low-cost improvement of crop yield.The seed coating agent provided by the application can appropriately prolong the activity of nitrogen-fixing bacteria, promote soil colonization, promote the growth and yield of crops and is suitable for low-cost improvement of crop yield.

Description

Technical Field

[0001] This invention belongs to the field of agricultural technology, and particularly relates to the field of microbial application technology, specifically a seed coating agent, its preparation method and its application. Background Technology

[0002] Nitrogen-fixing bacteria are active microorganisms capable of acquiring nitrogen from the air and converting it into nitrogen-containing substances that can be utilized by plants. Their nitrogen-fixing ability mainly relies on nitrogenase within their bodies. Studies have shown that the nitrogen content in the soil is negatively correlated with the activity of nitrogen-fixing bacteria in the soil, and excessive application of nitrogen fertilizer can inhibit the soil's biological nitrogen-fixing capacity. Furthermore, due to their unique nutritional pattern, the artificial fermentation enrichment rate of nitrogen-fixing bacteria is slow, and they require a live bacterial form to exercise their nitrogen-fixing ability, making it difficult to maintain their activity for long periods, resulting in low efficiency for large-scale production and utilization.

[0003] Corn is a crop that requires a large amount of nitrogen input to achieve high yields. It is highly adaptable and widely grown around the world. In areas where large-scale production inputs are difficult to obtain, its yield is limited by objective factors, and nitrogen-fixing bacteria have great practical potential in dealing with this situation. Summary of the Invention

[0004] The purpose of this invention is to address the problems existing in the prior art by providing a seed coating agent, its preparation method, and its application.

[0005] The objective of this invention is achieved through the following technical solution:

[0006] The first technical objective of this invention is to provide a seed coating agent, characterized in that the seed coating agent comprises: 1-5 parts by weight of thickener, 2-10 parts by weight of film-forming agent, 0.2-1 parts by weight of dye, 0.01-0.05 parts by weight of bittering agent, 5-25 parts by weight of formulation, and 100-1500 parts by weight of nitrogen-fixing bacteria solution.

[0007] The concentration of the nitrogen-fixing bacteria solution is 10. 6 ~10 9 The nitrogen-fixing bacteria culture was obtained by adding Ashube medium to a conventional fermenter with a concentration of CFU / ml.

[0008] The nitrogen-fixing bacteria in the nitrogen-fixing bacterial solution is *Azotobacter chrysophagus*.

[0009] One source of the nitrogen-fixing bacteria in the culture solution is the nitrogen-fixing bacteria from the China Agricultural Microbial Culture Collection Center, with the culture collection number ACCC10006.

[0010] The thickener is sodium alginate and gelatin, with a weight ratio of sodium alginate to gelatin of 2-5:1.

[0011] The film-forming agent is methylcellulose; the bittering agent is sucrose octaacetic acid ester.

[0012] The staining agents are iron oxide red and iron black, with a weight ratio of 2-5:1.

[0013] The formulation is a composition of mannitol, sucrose, ammonium nitrate, potassium dihydrogen phosphate, calcium carbonate, and magnesium sulfate, wherein the weight ratio of mannitol, sucrose, ammonium nitrate, potassium dihydrogen phosphate, calcium carbonate, and magnesium sulfate is 5-10:3-8:1-5:1-3:3-5:1-2.

[0014] The second technical objective of this invention is to provide a method for preparing a seed coating agent, characterized by the following steps: preparing 1-5 parts by weight of thickener, 2-10 parts by weight of film-forming agent, 0.2-1 parts by weight of dye, 0.01-0.05 parts by weight of bittering agent, 5-25 parts by weight of additives, and 100-1500 parts by weight of nitrogen-fixing bacteria solution in proportion; and mixing and stirring the nitrogen-fixing bacteria solution, thickener, dye, film-forming agent, bittering agent, and additives in a container in that order to obtain a seed coating agent containing nitrogen-fixing bacteria.

[0015] The third technical objective of this invention is to provide a method for using a seed coating agent, wherein the seed coating agent prepared for the second technical objective is mixed and stirred evenly with the seeds, and then air-dried for 30 to 60 minutes before sowing.

[0016] The fourth technical objective of this invention is to provide an application of a seed coating agent in prolonging the activity of nitrogen-fixing bacteria. In essence, it is a method for prolonging the survival time of nitrogen-fixing bacteria inside the seed coating agent. The seed coating agent with the above-mentioned formulation can extend the preservation time of the nitrogen-fixing bacteria contained therein to more than 20 days.

[0017] The fifth technical objective of this invention is to provide an application of a seed coating agent in promoting growth and increasing yield. The seed coating agent of the above formulation is mixed and stirred evenly with the seeds, dried in the shade, and then sown in low-fertility soil. During the verification, it was found that it could increase the biomass and thousand-grain weight of maize by 2% to 7%.

[0018] There are several indicators of low fertility. The "low fertility" referred to in this application only needs to meet one of the following indicators: soil organic matter less than 1%, soil total nitrogen less than 0.01%, and soil organic phosphorus content less than 15 mg / kg.

[0019] The present invention has the following advantages over the prior art:

[0020] The seed coating agent provided by this invention can appropriately prolong the activity of nitrogen-fixing bacteria, promote soil colonization, and promote crop growth and yield, making it suitable for increasing crop yield at low cost. Attached Figure Description

[0021] Appendix Figure 1 The effect of the seed coating agent provided by this invention on the emergence rate of maize;

[0022] Appendix Figure 2 The effect of the seed coating agent provided by this invention on the concentration of nitrogen-fixing bacteria in soil;

[0023] Appendix Figure 3 The effect of the seed coating agent provided by this invention on maize plant height at different time stages;

[0024] Appendix Figure 4 The effect of the seed coating agent provided by this invention on the stem diameter of maize at different time periods;

[0025] Appendix Figure 5 The effect of the seed coating agent provided by this invention on the root weight of maize at different time periods;

[0026] Appendix Figure 6 The effect of the seed coating agent provided by this invention on the plant weight of maize at different time periods;

[0027] Appendix Figure 7 The effect of the seed coating agent provided by this invention on the weight of maize ears;

[0028] Appendix Figure 8 The effect of the seed coating agent provided by this invention on the weight of maize ears;

[0029] Appendix Figure 9 The effect of the seed coating agent provided by this invention on the thousand-kernel weight of corn;

[0030] Appendix Figure 10 The effect of the seed coating agent provided by this invention on the weight of maize ears;

[0031] Appendix Figure 11 The effect of the seed coating agent provided by this invention on maize yield. Detailed Implementation

[0032] The technical solution of the present invention will now be clearly and completely described with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments.

[0033] This invention provides a seed coating agent comprising: 1-5 parts by weight of a thickener, 2-10 parts by weight of a film-forming agent, 0.2-1 parts by weight of a dye, 0.01-0.05 parts by weight of a bittering agent, 5-25 parts by weight of a compounding agent, and 100-1500 parts by weight of a nitrogen-fixing bacteria solution, wherein the preferred concentration of the nitrogen-fixing bacteria solution is 10. 6 ~10 9The nitrogen-fixing bacteria culture was obtained by adding Asbestos medium (Qingdao Haibo product number: HB8540) to a conventional fermenter; the nitrogen-fixing bacteria in the culture were preferably Azotobacter chrysotrichum.

[0034] In the above seed coating agent, the thickener is sodium alginate and gelatin, with a weight ratio of sodium alginate to gelatin of 2-5:1; the film-forming agent is methylcellulose, and the bittering agent is sucrose octaacetic acid ester; the dyeing agent is iron oxide red and iron black, with a weight ratio of iron oxide red and iron black of 2-5:1; the formulation is a combination of mannitol, sucrose, ammonium nitrate, potassium dihydrogen phosphate, calcium carbonate, and magnesium sulfate, with a weight ratio of mannitol, sucrose, ammonium nitrate, potassium dihydrogen phosphate, calcium carbonate, and magnesium sulfate of 5-10:3-8:1-5:1-3:3-5:1-2.

[0035] In the above seed coating agents, one of the sources of nitrogen-fixing bacteria in the nitrogen-fixing bacteria solution is the nitrogen-fixing bacteria with the strain preservation number ACCC10006 from the China Agricultural Microbial Culture Collection Center.

[0036] This invention provides a method for preparing a seed coating agent. The preparation method comprises the following steps: preparing 1-5 parts by weight of thickener, 2-10 parts by weight of film-forming agent, 0.2-1 parts by weight of dye, 0.01-0.05 parts by weight of bittering agent, 5-25 parts by weight of additives, and 100-1500 parts by weight of nitrogen-fixing bacteria solution in proportion; and mixing and stirring the nitrogen-fixing bacteria solution, thickener, dye, film-forming agent, bittering agent, and additives in a container in the order of the order of the nitrogen-fixing bacteria solution, thickener, dye, film-forming agent, bittering agent, and additives until homogeneous, thereby obtaining a seed coating agent containing nitrogen-fixing bacteria.

[0037] Mix the above-mentioned seed coating agent with the crop seeds, stir evenly, and let them air dry for 30 to 60 minutes before sowing.

[0038] This invention provides an application of a seed coating agent in prolonging the activity of nitrogen-fixing bacteria. The seed coating agent can extend the survival time of nitrogen-fixing bacteria inside the seed coating agent. Specifically, the seed coating agent of the above formulation can extend the preservation time of the nitrogen-fixing bacteria contained therein to more than 20 days.

[0039] This invention provides the application of a seed coating agent in promoting growth and increasing yield. The seed coating agent of the above formulation is mixed evenly with seeds, air-dried, and then sown in soils with low fertility (e.g., soil organic matter less than 1%; soil total nitrogen less than 0.01%; soil organic phosphorus content less than 15 mg / kg). The application of this seed coating agent can significantly promote crop growth and yield in low-fertility soils, but has no significant effect in fertile soils. It is more suitable for low-cost crop yield enhancement. During verification, it can increase maize biomass and thousand-grain weight by 2%–7%.

[0040] Example

[0041] A seed coating agent comprising: 1 part by weight of a thickener, 2 parts by weight of a film-forming agent, 0.2 parts by weight of a coloring agent, 0.01 parts by weight of a bittering agent, 5 parts by weight of a compounding agent, and 100 parts by weight of a concentration of 10. 8 The nitrogen-fixing bacteria solution contains CFU / ml of nitrogen-fixing bacteria, in which the weight ratio of sodium alginate to gelatin in the thickener is 5:1, and the weight ratio of iron oxide red to iron black in the dye is 2:1; the weight ratio of mannitol, sucrose, ammonium nitrate, potassium dihydrogen phosphate, calcium carbonate, and magnesium sulfate in the formulation is 6:4:3:1:4:1. The nitrogen-fixing bacteria in the nitrogen-fixing bacteria solution are sourced from the China Agricultural Microbial Culture Collection Center, with the culture collection number ACCC10006.

[0042] Preparation: Place the above-mentioned amounts of nitrogen-fixing bacteria solution, thickener, dye, film-forming agent, bittering agent, and other additives into a container and mix thoroughly to obtain a seed coating agent containing nitrogen-fixing bacteria.

[0043] Application: Mix the above seed coating agent with conventional corn seeds at a mass ratio of 1:30, stir evenly, and air dry for 30 minutes to obtain conventional corn seeds coated with the seed coating agent.

[0044] Experimental Example 1

[0045] Test plant: Potted maize, experimental variety: "Liangyu 99".

[0046] Experimental Methods: This experiment was conducted indoors in January 2023 using 100ml small pots. A control group (CK) and a seed coating agent treatment group (AC) were set up. The seed coating agent used in the seed coating agent treatment group was provided in the example. Four fertilizer gradients were set up: 100F (600 kg / ha), 90F (540 kg / ha), 80F (kg / ha), and 62.5F (kg / ha). Each treatment was replicated in triplicate, with 50 corn kernels per replicate. Germination rate was statistically analyzed.

[0047] Results Comparison: By Figure 1 It is known that the seed coating agent containing nitrogen-fixing bacteria provided by the present invention did not have a significant impact on the corn germination rate. Regardless of the multiple fertilizer gradients used, the germination rate was greater than 98%, and there was no significant difference between different treatments, indicating no adverse effects on corn.

[0048] Experimental Example 2

[0049] Test plant: Potted maize, experimental variety: "Liangyu 99".

[0050] Experimental Methods: Sowing was conducted on June 6th in both 2022 and 2023. A control group (CK) and a seed coating treatment group (AC) were established. The seed coating agent used in the seed coating treatment group was provided in the example. Four fertilizer gradients were set up: 100F (600 kg / ha), 90F (540 kg / ha), 80F (kg / ha), and 62.5F (kg / ha).

[0051] (1) Coated seeds (CK) without nitrogen-fixing bacteria, with other components the same as those in the above examples;

[0052] (2) The coated seeds provided in this embodiment of the invention (the number of live bacteria counted on the seed coating agent is 1.0 × 10⁻⁶) 8 (cfu / ml, and it is currently difficult to culture higher concentrations of bacterial solution);

[0053] (3) Each treatment is set up with 3 replicates, and each replicate is set to 30m. 2 The planting plots had a row spacing of 80cm and a plant spacing of 20cm. The corn emergence rate was surveyed from July 10, 2022 to July 14, 2023.

[0054] (4) A survey of corn growth was conducted on July 31, August 20, and September 10, 2022, and August 4, August 24, and September 13, 2023.

[0055] (5) Nitrogenase activity was measured in maize rhizosphere soil on September 10, 2022 and September 13, 2023.

[0056] (6) Maize yield was measured on September 30, 2022 and 2023.

[0057] Comparison results: The results are shown in Table 1.

[0058]

[0059] Table 1. Three-way ANOVA (F / P value) on the effects of fertilizer application (F, 100F represents 600 kg / ha fertilizer application), nitrogen-fixing bacteria coating (i), year (Y), and interaction on soil metabolic enzyme activity in different years.

[0060] Depend on Figure 1 It can be seen that the seed coating agent treatment for nitrogen-fixing bacteria did not have a significant effect on the emergence rate of maize.

[0061] Depend on Figure 2As shown in Table 1, changes in the amount of nitrogen-fixing bacteria seed coating agent and fertilizer application significantly affected soil nitrogenase activity, with the effect being more pronounced at lower fertilizer application rates. The nitrogen-fixing bacteria seed coating agent had a long-term interaction with maize, and the nitrogen-fixing bacteria in the seed coating agent successfully colonized in the soil. Year and various interactions did not have a significant impact. At a fertilizer application rate of 62.5F, soil nitrogenase activity increased by approximately 8%.

[0062] Depend on Figure 3 It can be seen that the seed coating agent treatment for nitrogen-fixing bacteria did not have a significant effect on the height of maize;

[0063] Depend on Figure 4 It can be seen that nitrogen-fixing bacteria seed coating agent has a significant effect on corn stalk diameter under reduced fertilizer application conditions. Under low fertilizer conditions (62.5F), nitrogen-fixing bacteria seed coating agent significantly increased corn stalk diameter (1%–15%).

[0064] Depend on Figure 5 It can be seen that nitrogen-fixing bacteria seed coating agent has a more significant effect on maize root weight under reduced fertilizer application conditions. Under low fertilizer conditions (62.5F), nitrogen-fixing bacteria seed coating agent significantly increased maize root weight (1%–56%).

[0065] Depend on Figure 6 It can be seen that nitrogen-fixing bacteria seed coating agent has a more significant effect on maize plant weight under reduced fertilizer application conditions. Under low fertilizer conditions (62.5F), nitrogen-fixing bacteria seed coating agent significantly increased maize plant weight (1%–17%).

[0066] Depend on Figure 7 It can be seen that the nitrogen-fixing bacteria seed coating agent has a significant effect on the ear weight of maize under the condition of reduced fertilizer application;

[0067] Depend on Figure 8 It can be seen that the nitrogen-fixing bacteria seed coating agent has a significant effect on the grain weight of maize ears under the condition of reduced fertilizer application;

[0068] Depend on Figure 9 It can be seen that nitrogen-fixing bacteria seed coating agents have a more significant effect on the thousand-kernel weight of maize under reduced fertilizer application conditions. Under low fertilizer conditions (62.5F), nitrogen-fixing bacteria seed coating agents significantly increased the thousand-kernel weight of maize (1%–7%). The main effect of nitrogen-fixing bacteria is to increase the thousand-kernel weight of maize, thus making each maize kernel more plump.

[0069] Depend on Figure 10 It can be seen that the nitrogen-fixing bacteria seed coating agent has a significant effect on the number of kernels per ear of maize under the condition of reduced fertilizer application;

[0070] Depend on Figure 11It can be seen that nitrogen-fixing bacteria seed coating agent has a more significant effect on maize yield under reduced fertilizer application conditions. Under low fertilizer conditions (62.5F), nitrogen-fixing bacteria seed coating agent significantly increased maize yield (1%–16%).

[0071] Experiment Example 3

[0072] Experimental method: The seed coating agent containing nitrogen-fixing bacteria and the original nitrogen-fixing bacteria solution provided in the examples were subjected to a culture medium plating and counting experiment every 10 days.

[0073] Comparison Results: As shown in Tables 2 and 3, the seed coating agent containing nitrogen-fixing bacteria significantly improved the preservation time of nitrogen-fixing bacteria activity at both 25℃ and 4℃. When the viable bacteria concentration was higher than 10... 6 Under the condition that CFU is an effective product, its shelf life can be extended by approximately 20 days. This provides favorable conditions for the preservation and transportation of seed coating agents.

[0074] viable bacteria count at 25℃ 0 days 10 days 20 days 30 days 40 days 50 days Original nitrogen-fixing bacteria culture 1.0*10^8 1.0*10^5 1.0*10^3 140 0 0 Seed coating agent 1.0*10^8 1.0*10^8 1.0*10^6 1.0*10^4 1.0*10^2 0

[0075] Table 2. Nitrogen-fixing bacteria activity of seed coating agents containing nitrogen-fixing bacteria and original nitrogen-fixing bacteria cultures at 25°C.

[0076] 4℃ / viable count 0 days 10 days 20 days 30 days 40 days 50 days Original nitrogen-fixing bacteria culture 1.0*10^8 1.0*10^8 1.0*10^6 1.0*10^6 1.0*10^5 1.0*10^4 Seed coating agent 1.0*10^8 1.0*10^8 1.0*10^8 1.0*10^7 1.0*10^7 1.0*10^6

[0077] Table 3. Nitrogen-fixing bacteria activity of seed coating agents containing nitrogen-fixing bacteria and original nitrogen-fixing bacteria cultures at 4°C.

[0078] The seed coating agent of the present invention can significantly promote crop growth and yield when applied to low-fertility soils, but has no significant effect on fertile soils. This seed coating agent is more suitable for increasing crop yield at a low cost.

[0079] The above embodiments are merely illustrative of the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the technical solutions of the embodiments of the present invention. All technologies not covered in this invention can be implemented using existing technologies.

Claims

1. Use of a seed coating agent for extending the activity of a nitrogen fixing bacteria, characterized in that: The seed coating agent can prolong the activity preservation time of the contained nitrogen-fixing bacteria to more than 20 days, and comprises 1-5 parts by weight of thickening agent, 2-10 parts by weight of film forming agent, 0.2-1 part by weight of dyeing agent, 0.01-0.05 part by weight of bittering agent, 5-25 parts by weight of agent, and 100-1500 parts by weight of nitrogen-fixing bacteria liquid; the agent is a combination of mannitol, sucrose, ammonium nitrate, potassium dihydrogen phosphate, calcium carbonate and magnesium sulfate, and the weight ratio of the mannitol, sucrose, ammonium nitrate, potassium dihydrogen phosphate, calcium carbonate and magnesium sulfate is 5-10:3-8:1-5:1-3:3-5:1-2; the nitrogen-fixing bacteria in the nitrogen-fixing bacteria liquid is round brown nitrogen-fixing bacteria; the seed coating agent is mixed with seeds and stirred uniformly, and then the seeds are sown after being dried in the shade.

2. Use of a seed coating agent according to claim 1 for extending the activity of diazotrophs, characterized in that: The concentration of the azotobacter bacterial solution is 10 6 ~ 10 9 CFU / ml, and the azotobacter bacterial solution is obtained by culturing the azotobacter in an ordinary fermentation tank with an asubel medium.

3. Use of the seed coating agent according to claim 1 for extending the activity of diazotrophs, characterized in that: The thickening agent is sodium alginate and gelatin, and the weight ratio of the sodium alginate and gelatin is 2-5:

1.

4. Use of the seed coating agent according to claim 1 for extending the activity of diazotrophs, characterized in that: The film forming agent is methyl cellulose; and the bittering agent is sucrose octaacetate.

5. Use of the seed coating agent according to claim 1 for extending the activity of nitrogen-fixing bacteria, characterized in that: The dyeing agent is iron oxide red and iron black, and the weight ratio of the iron oxide red and iron black is 2-5:

1.

6. Use of a seed coating agent according to any one of claims 1 to 5 for extending the activity of a nitrogen-fixing bacterium, characterized in that: The preparation method of the seed coating agent comprises the following steps: preparing 1-5 parts by weight of thickening agent, 2-10 parts by weight of film forming agent, 0.2-1 part by weight of dyeing agent, 0.01-0.05 part by weight of bittering agent, 5-25 parts by weight of agent, and 100-1500 parts by weight of nitrogen-fixing bacteria liquid according to the proportion; sequentially putting the nitrogen-fixing bacteria liquid, the thickening agent, the dyeing agent, the film forming agent, the bittering agent and the agent into a container and stirring uniformly to obtain the seed coating agent containing nitrogen-fixing bacteria; and mixing the seed coating agent with seeds and stirring uniformly, and then sowing the seeds after being dried in the shade.

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