Microbial complex microbial inoculant for promoting growth of poa pratensis as well as preparation method and application of microbial complex microbial inoculant
By using microbial complex agents of Bacillus subtilis and Bacillus licheniformis, the problem of slow growth rate of early-mature grassland grass is solved, significantly improving the growth rate of seedlings and plant high growth rate, simplifying the planting process and providing high-quality seedling sources.
Patent Information
- Application Number
- CN202311601996.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-11-28
- Publication Date
- 2025-05-30
AI Technical Summary
The slow growth rate of grass early-mature grass grass is caused by a lot of time during the planting process. In addition, the number of grass early-mature grass grass varieties in my country is relatively small and lacks high-quality species.
A microbial complex bacterial agent is provided, including Bacillus subtilis and Bacillus licheniformis, with a ratio of 1:1, prepared by solid fermentation medium for the promotion of seedling growth of early grass grass.
Significantly improve the growth rate of early-mature grassland in the seedling stage, improve the plant height growth rate, and enable the seedlings to meet the transplanting standards faster, simplify the planting process and reduce time costs.
Smart Images
Figure CN120059989A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of plant biotechnology, and particularly relates to a microbial complex bactericide for promoting the growth of Poa pratensis, a preparation method thereof, and an application thereof. Background Art
[0002] As a commonly used cool-season turfgrass species in northern China, Poa pratensis has the characteristics of fast turf establishment speed, soft leaves, dark green leaf color, and long sustainable green period. However, at present, the number of Poa pratensis varieties in China is small, and there are only 9 varieties registered and approved by the National Turfgrass Variety Approval Committee. Most of the varieties that can be used for turf establishment are introduced varieties. Carrying out the breeding work of Poa pratensis varieties can provide high-quality seed sources for turf establishment in northern China. However, in the breeding work of Poa pratensis varieties, it is necessary to repeatedly plant Poa pratensis. Because the growth rate of Poa pratensis is slow, a large amount of time is often required in the planting process. Therefore, it is necessary to provide a microbial complex bactericide for promoting the growth of Poa pratensis. Summary of the Invention
[0003] To solve the above technical problems, the present invention provides a microbial complex bactericide for promoting the growth of Poa pratensis to improve the growth rate of Poa pratensis in the seedling stage. When the microbial complex bactericide of the present invention is used in the maintenance of Poa pratensis seedlings, the plant height growth rate of Poa pratensis is significantly increased, and the seedlings can reach the transplanting standard faster.
[0004] A microbial complex bactericide for promoting the growth of Poa pratensis, comprising Bacillus subtilis and Bacillus licheniformis, and the ratio of Bacillus subtilis to Bacillus licheniformis is Bacillus subtilis: Bacillus licheniformis = 1:1.
[0005] The preparation method of the microbial complex bactericide is: after separately preparing Bacillus subtilis and Bacillus licheniformis, they are mixed in a ratio of 1:1.
[0006] Moreover, the preparation method of Bacillus subtilis is to inoculate a Bacillus subtilis strain into a solid fermentation medium for fermentation culture; The preparation method of Bacillus licheniformis is to inoculate a Bacillus licheniformis strain into a solid fermentation medium for fermentation culture.
[0007] Moreover, the temperature of fermentation culture is 20 - 25 °C, and the fermentation time is 30 - 35 d.
[0008] Moreover, the solid fermentation medium is composed of a substrate, corn, soybean meal, lime, gypsum, and sugar.
[0009] Moreover, in the solid fermentation medium, the mass of the substrate is 4000 - 4600 g, the mass of the corn is 450 - 550 g, the mass of the soybean meal is 45 - 55 g, the mass of the lime is 4 - 6 g, the mass of the gypsum is 4 - 6 g, and the mass of the sugar is 8 - 12 g.
[0010] Moreover, in the solid fermentation medium, the mass of the substrate is 4300 g, the mass of the corn is 500 g, the mass of the soybean meal is 50 g, the mass of the lime is 5 g, the mass of the gypsum is 5 g, and the mass of the sugar is 10 g.
[0011] Moreover, the substrate is 90 - 95% nutrient soil and 5 - 10% ordinary soil, and the water content of the substrate is 35 - 40%.
[0012] Applying the above microbial complex bactericide to the seeding and establishment of Poa pratensis, it is characterized by including the following steps: S1. Pretreatment of Poa pratensis seeds before sowing: Use a rice milling machine to remove the fluff on the surface of Poa pratensis seeds; S2. Seeding and establishment of Poa pratensis: After treatment, sow the Poa pratensis seeds in rows in plots. The plot area is 2 m × 5 m, the row spacing is 35 - 45 cm, the sowing depth is 0.5 - 1 cm, cover the soil and roll it. Apply the microbial complex bactericide 28 - 32 days after emergence, and the dosage is 450 - 550 g per plot.
[0013] Moreover, the sand roller of the rice milling machine is 100 - mesh and the long holes of the sieve plate are 0.6 mm; the application method of the microbial complex bactericide is ditch broadcasting.
[0014] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. The present invention provides a microbial complex bactericide for promoting the growth of Poa pratensis, including Bacillus subtilis and Bacillus licheniformis. The ratio of Bacillus subtilis to Bacillus licheniformis is Bacillus subtilis: Bacillus licheniformis = 1:1; this microbial complex bactericide can improve the growth rate of Poa pratensis in the seedling stage. When using the microbial complex bactericide of the present invention in the maintenance of Poa pratensis seedlings, the plant height growth rate of Poa pratensis is significantly increased, enabling the seedlings to reach the transplanting standard faster; 2. The present invention provides a preparation method of the microbial complex bactericide and provides a technical solution for the seeding and establishment of Poa pratensis, with simple operation and convenient for large - scale promotion. Description of the Drawings
[0015] Figure 1 It is the growth trend diagram of Poa pratensis in treatment 5 in Experiment 2 of the present invention; Figure 2 It is the growth trend diagram of Poa pratensis in treatment 4 in Experiment 2 of the present invention; Figure 3 It is the growth diagram of Kentucky bluegrass in the CK group in Experiment 2 of the present invention; Figure 4 It is the production process diagram of the solid fermentation medium of the present invention; Figure 5 It is the finished product diagram of the solid fermentation medium of the present invention; Figure 6 It is the growth condition of Bacillus licheniformis; Figure 7 It is the growth condition of Azotobacter chroococcum. Detailed implementation manners Example 1
[0016] A microbial complex bactericide for promoting the growth of Kentucky bluegrass, comprising Bacillus subtilis and Bacillus licheniformis, and the ratio of Bacillus subtilis to Bacillus licheniformis is Bacillus subtilis: Bacillus licheniformis = 1:1.
[0017] The preparation method of the microbial complex bactericide is: after separately preparing Bacillus subtilis and Bacillus licheniformis, mix them in a ratio of 1:1.
[0018] Further, the preparation method of Bacillus subtilis is to inoculate a Bacillus subtilis strain into a solid fermentation medium for fermentation culture.
[0019] The preparation method of Bacillus licheniformis is to inoculate a Bacillus licheniformis strain into a solid fermentation medium for fermentation culture.
[0020] Further, the temperature of fermentation culture is 22 °C, and the fermentation time is 30 d.
[0021] Further, the solid fermentation medium is composed of substrate, corn, soybean meal, lime, gypsum, and sugar.
[0022] Further, in the solid fermentation medium, the mass of the substrate is 4000 g, the mass of the corn is 450 g, the mass of the soybean meal is 45 g, the mass of the lime is 4 g, the mass of the gypsum is 4 g, and the mass of the sugar is 8 g.
[0023] Further, the substrate is 90% nutrient soil and 10% ordinary soil, and the water content of the substrate is 35%. Example 2
[0024] A microbial complex bactericide for promoting the growth of Kentucky bluegrass, comprising Bacillus subtilis and Bacillus licheniformis, and the ratio of Bacillus subtilis to Bacillus licheniformis is Bacillus subtilis: Bacillus licheniformis = 1:1.
[0025] The preparation method of the microbial complex bactericide is as follows: After separately preparing Bacillus subtilis and Bacillus licheniformis, they are mixed in a ratio of 1:1.
[0026] Further, the preparation method of the Bacillus subtilis is to inoculate the Bacillus subtilis strain into a solid fermentation medium for fermentation culture; The preparation method of the Bacillus licheniformis is to inoculate the Bacillus licheniformis strain into a solid fermentation medium for fermentation culture.
[0027] Further, the temperature of the fermentation culture is 25 °C, and the fermentation time is 35 d.
[0028] Further, the solid fermentation medium is composed of substrate, corn, soybean meal, lime, gypsum, and sugar.
[0029] Further, in the solid fermentation medium, the mass of the substrate is 4600 g, the mass of the corn is 550 g, the mass of the soybean meal is 55 g, the mass of the lime is 6 g, the mass of the gypsum is 6 g, and the mass of the sugar is 12 g.
[0030] Further, the substrate is 95% nutrient soil and 5% ordinary soil, and the water content of the substrate is 40%. Example 3
[0031] A microbial complex bactericide for promoting the growth of Poa pratensis L. includes Bacillus subtilis and Bacillus licheniformis, and the ratio of the Bacillus subtilis to the Bacillus licheniformis is Bacillus subtilis: Bacillus licheniformis = 1:1.
[0032] The preparation method of the microbial complex bactericide is as follows: After separately preparing Bacillus subtilis and Bacillus licheniformis, they are mixed in a ratio of 1:1.
[0033] Further, the preparation method of the Bacillus subtilis is to inoculate the Bacillus subtilis strain into a solid fermentation medium for fermentation culture; The preparation method of the Bacillus licheniformis is to inoculate the Bacillus licheniformis strain into a solid fermentation medium for fermentation culture.
[0034] Further, the temperature of the fermentation culture is 22 °C, and the fermentation time is 33 d.
[0035] Further, the solid fermentation medium is composed of substrate, corn, soybean meal, lime, gypsum, and sugar.
[0036] Further, in the solid fermentation medium, the mass of the substrate is 4300 g, the mass of the corn is 500 g, the mass of the soybean meal is 50 g, the mass of the lime is 5 g, the mass of the gypsum is 5 g, and the mass of the sugar is 10 g.
[0037] Furthermore, the substrate is 92% nutrient soil and 8% ordinary soil, and the water content of the substrate is 38%. Example 4
[0038] Apply the microbial complex agent for promoting the growth of Kentucky bluegrass prepared in Example 3 to the seeding and establishment of Kentucky bluegrass, including the following steps: S1. Pretreatment of Kentucky bluegrass seeds before sowing: Use a rice milling machine to remove the fluff on the surface of Kentucky bluegrass seeds; S2. Seeding and establishment of Kentucky bluegrass: After treatment, sow the Kentucky bluegrass seeds in rows in plots. The plot area is 2m×5m, the row spacing is 35cm, the sowing depth is 0.5cm, cover the soil and roll it. Apply the microbial complex agent 28 days after emergence, and the dosage is 450g per plot.
[0039] Furthermore, the sand roller of the rice milling machine is 100 mesh and the long holes of the sieve plate are 0.6mm; the application method of the microbial complex agent is to spread it by opening a trench. Example 5
[0040] Apply the microbial complex agent for promoting the growth of Kentucky bluegrass prepared in Example 3 to the seeding and establishment of Kentucky bluegrass, including the following steps: S1. Pretreatment of Kentucky bluegrass seeds before sowing: Use a rice milling machine to remove the fluff on the surface of Kentucky bluegrass seeds; S2. Seeding and establishment of Kentucky bluegrass: After treatment, sow the Kentucky bluegrass seeds in rows in plots. The plot area is 2m×5m, the row spacing is 45cm, the sowing depth is 1cm, cover the soil and roll it. Apply the microbial complex agent 32 days after emergence, and the dosage is 550g per plot.
[0041] Furthermore, the sand roller of the rice milling machine is 100 mesh and the long holes of the sieve plate are 0.6mm; the application method of the microbial complex agent is to spread it by opening a trench. Example 6
[0042] Apply the microbial complex agent for promoting the growth of Kentucky bluegrass prepared in Example 3 to the seeding and establishment of Kentucky bluegrass, including the following steps: S1. Pretreatment of Kentucky bluegrass seeds before sowing: Use a rice milling machine to remove the fluff on the surface of Kentucky bluegrass seeds; S2. Seeding and establishment of Kentucky bluegrass: After treatment, sow the Kentucky bluegrass seeds in rows in plots. The plot area is 2m×5m, the row spacing is 40cm, the sowing depth is 0.8cm, cover the soil and roll it. Apply the microbial complex agent 30 days after emergence, and the dosage is 500g per plot.
[0043] Further, the sand roller of the rice milling machine is 100 mesh, and the long holes of the sieve plate are 0.6 mm; the application method of the microbial complex bactericide is ditch spreading. Experimental part Experiment 1
[0044] Six strains of Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus pumilus, Azotobacter chroococcum and Bacillus jiaodongensis were cultivated respectively. The specific cultivation methods are as follows: The six strains of Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus pumilus, Azotobacter chroococcum and Bacillus jiaodongensis were respectively inoculated into the solid fermentation medium and fermented and cultured for 33 days at 22 °C.
[0045] Among them, the solid fermentation medium used for cultivating Azotobacter chroococcum, Bacillus licheniformis, Bacillus subtilis and Bacillus jiaodongensis is the solid fermentation medium used in Example 3, specifically: 4300 g of substrate, 500 g of corn, 50 g of soybean meal, 5 g of lime, 5 g of gypsum, and 10 g of sugar.
[0046] The solid fermentation medium used for cultivating Bacillus amyloliquefaciens is 4300 g of substrate, 500 g of corn, 50 g of soybean meal, 5 g of lime, 5 g of gypsum, 10 g of sugar, and 250 g of wheat bran.
[0047] The solid fermentation medium used for cultivating Bacillus pumilus is 4300 g of substrate, 500 g of corn, 50 g of soybean meal, 5 g of lime, 5 g of gypsum, 10 g of sugar, and 300 g of wheat bran. Experiment 2
[0048] Through the antagonism test of different strains, the strains that can coexist were selected; the results of the antagonism test are shown in Table 1 below, where * represents weak antagonism, ** represents sub-strong antagonism, and *** represents strong antagonism.
[0049] Table 1 Results of antagonism test
[0050] According to the results of the antagonism test, the combinations and proportions of different strains were set, and the CK group was also set, as shown in Table 2 specifically, where 1 part represents 250 g of the solid fermentation bactericide of a single strain, and after being fully mixed according to different proportions, it was applied to the rhizosphere of plants.
[0051] Table 2 Proportions of different microbial bactericides CK Blank control Treatment 1 Bacillus subtilis: Bacillus licheniformis: Azotobacter chroococcum = 1:1:2 Treatment 2 Bacillus subtilis: Bacillus licheniformis: Bacillus amyloliquefaciens = 1:1:2 Treatment 3 Bacillus subtilis: Bacillus licheniformis: Bacillus mucilaginosus = 1:1:2 Treatment 4 Bacillus subtilis: Bacillus licheniformis: Bacillus pumilus = 1:1:2 Treatment 5 Bacillus subtilis: Bacillus licheniformis = 1:1 After applying the microbial bactericide, the plant height was observed once every 7 days for a total of 4 times. The variance analysis of the plant height of different treatments was carried out to compare the significance of differences between different treatments, as shown in Table 3 specifically.
[0052] Table 3 Plant height changes under different treatment conditions (day / month) Treatment 22 / 7 29 / 7 5 / 8 12 / 8 Average growth rate (cm / d) CK 7.29 cm 9.26 cm 11.93 cm 12.73 cm 0.26 Treatment 1 8.85 cm 9.02 cm 10.61 cm 11.07 cm 0.11 Treatment 2 10.40 cm 12.90 cm 12.68 cm 13.62 cm 0.15 Treatment 3 11.09cm 11.99 cm 12.30 cm 13.62 cm 0.11 Treatment 4 9.88 cm 13.34 cm 13.83 cm 16.36 cm 0.31 Treatment 5 11.20 cm 13.89 cm 15.45 cm 21.27 cm 0.48 As can be seen from Table 3, on July 22, the plant heights of Treatments 1-5 were all higher than those of the CK group. The plant height of Treatment 5 was the highest, reaching 11.20 cm. On July 29, the CK group was slightly higher than Treatment 1, and Treatments 2-5 were all higher than the CK group. The plant height of Treatment 5 was the highest, reaching 13.89 cm. On August 5, the CK group was higher than Treatment 1, and Treatments 2-5 were all higher than the CK group. The plant height of Treatment 5 was the highest, reaching 15.45 cm. On August 12, the plant height of the CK was 12.73 cm, and the plant heights of Treatments 1, 2, 3, 4, and 5 were 11.07 cm, 13.62 cm, 13.62 cm, 16.36 cm, and 21.27 cm respectively. The CK group (12.73 cm) was higher than Treatment 1 (11.07 cm) and lower than Treatments 2-4. The plant height of Treatment 5 was the highest, reaching 21.27 cm, significantly higher than other treatments.
[0053] In terms of the average growth rate, the average growth rate of the CK was 0.26 cm / d, and the average growth rates of Treatments 1, 2, 3, 4, and 5 were 0.11 cm / d, 0.15 cm / d, 0.11 cm / d, 0.31 cm / d, and 0.48 cm / d respectively. The order of the average growth rate was: Treatment 5 > Treatment 4 > CK > Treatment 2 > Treatment 1 > Treatment 3.
[0054] In conclusion, Treatment 5 adopted the technical solution of the present invention, which had the most obvious growth promotion effect on Poa pratensis L. It could be proved that the technical solution of the present invention was the best technical solution.
Claims
1. A microbial complex bactericide for promoting the growth of Kentucky bluegrass, characterized in that, the microbial complex bactericide includes Bacillus subtilis and Bacillus licheniformis, and the ratio of Bacillus subtilis to Bacillus licheniformis is Bacillus subtilis: Bacillus licheniformis = 1:
1.
2. The microbial complex bactericide for promoting the growth of Kentucky bluegrass according to claim 1, characterized in that, the preparation method of the microbial complex bactericide is: after separately preparing Bacillus subtilis and Bacillus licheniformis, mix them in a ratio of 1:
1.
3. The microbial complex bactericide for promoting the growth of Kentucky bluegrass according to claim 2, characterized in that, the preparation method of the Bacillus subtilis is to inoculate the Bacillus subtilis strain into a solid fermentation medium for fermentation culture; the preparation method of the Bacillus licheniformis is to inoculate the Bacillus licheniformis strain into a solid fermentation medium for fermentation culture.
4. The microbial complex bactericide for promoting the growth of Kentucky bluegrass according to claim 3, characterized in that, the temperature of fermentation culture is 20 - 25 °C, and the fermentation time is 30 - 35 d.
5. The microbial complex bactericide for promoting the growth of Kentucky bluegrass according to claim 3, characterized in that, the solid fermentation medium is composed of substrate, corn, soybean meal, lime, gypsum, and sugar.
6. The microbial complex bactericide for promoting the growth of Kentucky bluegrass according to claim 5, characterized in that, in the solid fermentation medium, the mass of the substrate is 4000 - 4600 g, the mass of the corn is 450 - 550 g, the mass of the soybean meal is 45 - 55 g, the mass of the lime is 4 - 6 g, the mass of the gypsum is 4 - 6 g, and the mass of the sugar is 8 - 12 g.
7. The microbial complex bactericide for promoting the growth of Kentucky bluegrass according to claim 6, characterized in that, in the solid fermentation medium, the mass of the substrate is 4300 g, the mass of the corn is 500 g, the mass of the soybean meal is 50 g, the mass of the lime is 5 g, the mass of the gypsum is 5 g, and the mass of the sugar is 10 g.
8. The microbial complex bactericide for promoting the growth of Kentucky bluegrass according to claim 5, characterized in that, the substrate is 90 - 95% nutrient soil and 5 - 10% ordinary soil, and the water content of the substrate is 35 - 40%.
9. Applying the microbial complex bactericide for promoting the growth of Kentucky bluegrass according to any one of claims 1 - 8 to the sowing and establishment of Kentucky bluegrass, characterized in that, it includes the following steps: S1. Pretreatment of Kentucky bluegrass seeds before sowing: Use a rice milling machine to remove the fluff on the surface of Kentucky bluegrass seeds; S2. Sowing and establishment of Kentucky bluegrass: After treatment, sow the Kentucky bluegrass seeds in rows in plots, the plot area is 2 m × 5 m, the row spacing is 35 - 45 cm, the sowing depth is 0.5 - 1 cm, cover the soil and compact it, and apply the microbial complex bactericide 28 - 32 d after emergence, with a dosage of 450 - 550 g per plot.
10. The application of the microbial complex bactericide for promoting the growth of Kentucky bluegrass according to claim 9, characterized in that, The sand roller of the rice mill is 100 mesh, and the long holes of the sieve plate are 0.6 mm; the application method of the microbial complex bactericide is ditch spreading.