Soil microorganism for promoting growth of tea trees and improving flavor of tea leaves

Abstract

The invention belongs to the technical field of microorganisms, and discloses a soil microorganism for promoting the growth of tea trees and improving the quality of tea leaves, the strain name is Streptomyces cerevisiae SI-3B1, the preservation unit is Guangdong Microbial Culture Collection Center, the preservation date is September 13, 2024, and the preservation number is GDMCC No.65129. The invention further discloses a preparation method of the soil microorganism for promoting the growth of the tea trees and improving the quality of the tea leaves. The microorganisms effectively improve the content of theanine and water extract of fresh leaves of one bud and two leaves of large-leaf spring tea, summer tea leaves are thicker, and the biomass of the leaves is increased.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and specifically relates to soil microorganisms for promoting the growth of tea trees and improving the quality of tea leaves. Background Art

[0002] The rhizosphere microenvironment of tea trees refers to the area where tea tree soil, tea tree roots and soil microorganisms are closely integrated and interact with each other. This area is the main place for tea trees to absorb nutrients and grow and develop, and a good rhizosphere microenvironment can promote the healthy growth of tea trees and improve the quality of tea. Tea tree soil microorganisms mainly include bacteria, archaea, fungi and actinomycetes. Due to the differences in tea tree growth altitude, soil type, nutrient status, tea tree variety, tree age and growing season, the species composition of rhizosphere microorganisms is also different. Studies have shown that tea rhizosphere microorganisms play an important role in the cultivation and quality formation of tea.

[0003] At present, the types of probiotics commonly used in tea soil conditioning include Bacillus subtilis, Bacillus megaterium, Bacillus thuringiensis, Bacillus licheniformis, Bacillus laterosporus, Bacillus gelatinous, Streptomyces jingyangensis, brown nitrogen-fixing bacteria and lactic acid bacteria. These strains are selected and studied in order to improve tea soil and increase agricultural output. Although there are a large number of soil beneficial microbial preparations or fertilizers on the market, most of them are based on Bacillus. However, the original ecological environment of many strains is not from the soil. Because microorganisms have a certain selectivity for the ecological environment, there are problems such as weak adaptability and unclear effect. For specific tea varieties, different growth conditions and geographical environments, screening, functional definition and applicable environment research of native beneficial bacteria in the soil has always been the focus of agricultural production. Actinomycetes are an important indigenous microbial group in the soil.

[0004] Soil microorganisms of the Actinomycete family are known for their unique ability to synthesize secondary metabolites with significant biological activity. They have the following advantages as agricultural microorganisms: (1) They show good effects against various plant pathogens. They can produce antibiotics and other bioactive compounds that inhibit the growth of harmful fungi, bacteria and nematodes. Protect tea plants from diseases. (2) They promote plant growth by producing growth-promoting substances such as plant hormones (coenzymes, cytokinins and gibberellins) and enzymes. These compounds can stimulate root development, nutrient absorption and overall plant health. They can improve plant growth and yield. (3) They have the potential to fix atmospheric nitrogen and convert it into a form that can be assimilated by plants. Tea plants have a high demand for nitrogen, and the presence of nitrogen-fixing bacteria in the soil can promote plant nitrogen demand and increase soil fertility. (4) They play an important role in soil ecology by decomposing plant materials and recycling nutrients. They can decompose organic matter and release nutrients in a form that can be used by plants. Improve soil structure, nutrient cycling and overall soil health.

[0005] In terms of tea plantations, the application of soil-derived actinomycetes can improve nutrient absorption in the rhizosphere of tea trees, increase soil fertility, promote root development and inhibit plant diseases, which has a positive impact on the taste and quality of tea. Summary of the invention

[0006] The purpose of the present invention is to provide a soil probiotic actinomycete for promoting the growth of tea trees and improving the quality of tea leaves, the strain name: Streptomyces cerasinus SI-3B1, the deposit unit: Guangdong Microbial Culture Collection Center, the deposit date: September 13, 2024, the deposit number: GDMCC No. 65129. The strain was isolated from the native soil of Yunnan large-leaf arbor in Xishuangbanna Dai Autonomous Prefecture, Yunnan Province.

[0007] Physical and chemical properties: The strain is heterotrophic and aerobic. After 7 days of culture on glycerol asparagine agar (ISP5) medium, the colonies are white, round, convex and firm, with a dry surface and neat edges (Appendix 2). There are well-developed aerial branched hyphae without transverse septa. The spores are oval and arranged in spiral spore chains. When cultured on Streptomyces medium No. 2 (ISP2), the edges of the colonies are smooth, the surface is dry, and the aerial hyphae are yellow. When cultured on casein mineral medium (0.75NaCl, 1.75K2HPO4, 0.25MgSO4·7H2O, 0.055CaCl2, 0.010FeSO4·7H20, 0.005ZnSO4·7H2O, 10 casein, g·L-1), the edges of the colonies are smooth, the surface is dry, the aerial hyphae are grayish white, and no soluble pigment is secreted. The strain has the ability to fix nitrogen and secrete iron carriers. The results of sequencing the 16s rDNA region of the strain using primers 27F and 1492R showed that the homology with the Streptomyces cerasinus SR3-134 strain in the NCBI database reached 100%.

[0008] The present invention relates to a soil microorganism for promoting the growth of tea trees and improving the quality of tea leaves. The soil microorganism is taxonomically named Streptomyces cerasinus SI-3B1, and the deposit unit is Guangdong Microbiological Culture Collection Center, the deposit date is September 10, 2024, and the deposit number is GDMCC No.65129.

[0009] The present invention also relates to a bacterial agent comprising the soil microorganism according to claim 1.

[0010] Preferably, the bacterial agent also includes auxiliary materials.

[0011] More preferably, the auxiliary material is selected from soil or light calcium carbonate or a mixture of the two.

[0012] More preferably, the bacterial agent is prepared according to the following method:

[0013] The fermentation culture solution of the soil microorganisms according to claim 1, soil and light calcium carbonate are mixed in a certain proportion and cultured to obtain a bacterial agent.

[0014] Most preferably, the bacterial agent is prepared according to the following method:

[0015] The fermentation culture solution of the soil microorganisms according to claim 1, soil and light calcium carbonate are mixed in a ratio of 1-5:10-30:0.1-0.5 (v / w / w), and cultured for 48 hours at a temperature of 30° C. and a humidity of 80% to obtain a bacterial agent.

[0016] The present invention also relates to the application of soil microorganisms and microbial agents in promoting the growth of tea trees and improving the quality of tea leaves.

[0017] Preferably, the application comprises: applying the soil microorganisms according to claim 1 or the bacterial agents according to claims 2-6 to the soil at the root of the tea tree at a depth of 10-40 cm. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 :The colony picture of the strain after 7 days of culture on the special medium for actinomycetes is shown in Figure 1 .

[0019] The beneficial effects achieved by the present invention mainly include but are not limited to the following aspects:

[0020] According to the results of the test and determination, the strain of the present invention can increase the carbon source available in the soil by increasing the activity of amylase and cellulase in the soil, and can increase the nitrogen source available in the soil by increasing the activity of urease in the soil. These changes are conducive to promoting the growth of tea trees and increasing the accumulation of flavor substances. Applying the bacterial agent 20-25 days before the spring tea is picked can effectively increase the content of amino acids and water extracts in the fresh tea leaves with one bud and two leaves just unfolding. In addition, using the bacterial agent 45 days before picking can make the summer tea leaves thicker and increase the biomass of the leaves. It has a positive effect on improving the quality and yield of tea leaves. DETAILED DESCRIPTION

[0021] Those skilled in the art can refer to the content of this article and appropriately improve the process parameters. It is particularly important to point out that all similar replacements and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The products and methods of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the products and methods described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention. In order to further understand the present invention, the present invention is described in detail below in conjunction with embodiments.

[0022] Example 1

[0023] 1. Activation of strains: 1L of liquid culture medium (10g of malt extract, 4g of yeast extract, 4g of D-glucose, 1000mL of distilled water, adjusted to pH = 7.2, autoclaved), inoculated with 100mg of freeze-dried powder (1.0×10 6 CFU / g), at 25°C, with an aeration rate of 0.5 vvm and stirring at 200 rpm for 72 h to obtain the strain culture solution.

[0024] 2. Preparation of the microbial agent: Mix the culture solution, autoclaved tea garden soil and light calcium carbonate in a ratio of 1:20:0.2 (v / w / w, ml / g / g). Cultivate for 48 hours at a temperature of 30°C and a humidity of 80% to obtain the microbial agent.

[0025] 3. Use of microbial agent: 20 days before picking spring tea, apply 250g of microbial agent to the soil 15cm deep from the roots of large-leaf shrub tea trees for each tea tree; 30 days before picking summer tea, apply 400g of microbial agent to the soil 15cm deep from the roots of tea trees for each tea tree.

[0026] 4. Index detection and results: The tea trees in the same tea garden that were treated with sterilized soil + calcium carbonate of the same mass were used as controls. Fifty fresh leaves were randomly picked from each tree. Theanine and tea water extract in tea were determined using GB / T 23193-2017 and GB / T8305-2013 methods, respectively, and the leaf mass was determined by weighing. Soil samples were taken from 15 cm deep soil near the rhizosphere, and 3 sampling points were taken for each tea tree. 50 g of soil was collected from each sampling point and mixed. A total of 15 tea trees were randomly sampled for analysis. The activity of urease was determined using the sodium phenol-sodium hypochlorite colorimetric method, with 1 gram of soil filtrate catalyzing the generation of 1 μg NH3-N in 1 minute representing 1 U / g. The activities of amylase and cellulase were determined by 3,5-dinitrosalicylic acid colorimetry using 1% soluble starch and 1% carboxymethyl cellulose solution as substrates, respectively. 1 U / g was defined as the amount of glucose produced per minute by catalysis of 1 μg of soil filtrate.

[0027] The measurement results of relevant indicators are shown in Table 1 below.

[0028] Table 1

[0029]

[0030]

[0031] Compared with the control, the activities of urease, amylase and cellulase in the soil treated with the bacterial agent prepared by the strain of the present invention were increased by 23.8%, 21.1% and 44.0%, respectively. The theanine content of fresh leaves of spring tea of ​​tea trees increased by 6.9%, the water extract content increased by 10.0%, and the weight of fresh leaves of summer tea increased by 5.1%.

[0032] Example 2

[0033] 1. Activation of strains: 1L of liquid culture medium (10g of malt extract, 4g of yeast extract, 4g of D-glucose, 1000mL of distilled water, adjusted to pH = 7.2, autoclaved), inoculated with 120mg of freeze-dried powder (1.0×10 6CFU / g), at 25°C, with an aeration rate of 0.5 vvm and stirring at 200 rpm for 72 h to obtain the culture solution.

[0034] 2. Preparation of the microbial agent: Mix the culture solution, autoclaved tea garden soil and light calcium carbonate in a ratio of 1:25:0.4 (v / w / w). Cultivate for 48 hours at a temperature of 30°C and a humidity of 80% to obtain the microbial agent.

[0035] 3. Use of microbial agent: 20 days before picking spring tea, apply 300g of microbial agent to the soil 20cm deep from the roots of large-leaf shrub tea trees for each tea tree; 30 days before picking summer tea, apply 500g of microbial agent to the soil 20cm deep from the roots of tea trees for each tea tree.

[0036] 4. Index detection and results: The tea trees in the same tea garden that were treated with sterilized soil + calcium carbonate of the same mass were used as controls. 50 fresh leaves were randomly picked from each tree. Theanine and tea water extract in tea were determined using GB / T 23193-2017 and GB / T8305-2013 methods, respectively, and the leaf mass was determined by weighing. Soil samples were taken from 20 cm deep soil near the rhizosphere, and 3 sampling points were taken for each tea tree. 50 g of soil was collected from each sampling point and mixed. A total of 15 tea trees were randomly sampled for analysis. The activity of urease was determined using the sodium phenol-sodium hypochlorite colorimetric method, with 1 μg of NH3-N catalyzed by 1 gram of soil filtrate representing 1 U / g. The activities of amylase and cellulase were determined by 3,5-dinitrosalicylic acid colorimetry using 1% soluble starch and 1% carboxymethyl cellulose solution as substrates, respectively. 1 U / g was defined as the amount of glucose produced per minute by catalysis of 1 μg of soil filtrate.

[0037] The measurement results of relevant indicators are shown in Table 2 below.

[0038] Table 2

[0039]

[0040]

[0041] Compared with the control, the activities of urease, amylase and cellulase in the soil treated with the bacterial agent prepared by the strain of the present invention were increased by 47.6%, 24.3% and 64.3% respectively. The theanine content of fresh leaves of spring tea of ​​tea trees increased by 22.4%, and the water extract content increased by 13.7%. The weight of fresh leaves of summer tea increased by 7.2%.

[0042] Example 3

[0043] 1. Activation of strains: 1L of liquid culture medium (10g of malt extract, 4g of yeast extract, 4g of D-glucose, 1000mL of distilled water, adjusted to pH = 7.2, autoclaved), inoculated with 150mg of freeze-dried powder (1.0×10 6 CFU / g), at 25°C, with an aeration rate of 0.5 vvm and stirring at 200 rpm for 72 h to obtain the culture solution.

[0044] 2. Preparation of the microbial agent: Mix the culture solution, autoclaved tea garden soil and light calcium carbonate in a ratio of 1:30:0.3 (v / w / w), and culture for 48 hours at a temperature of 30°C and a humidity of 80% to obtain the microbial agent.

[0045] 3. Use of microbial agents: 20 days before picking spring tea, apply 200g of microbial agent to the soil 15cm deep from the roots of large-leaf shrub tea trees for each tea tree; 30 days before picking summer tea, apply 300g of activated compound microbial agent to the soil 15cm deep from the roots of tea trees for each tea tree.

[0046] 4. Index detection and results: The tea trees in the same tea garden that were treated with sterilized soil + calcium carbonate of the same mass were used as controls. Fifty fresh leaves were randomly picked from each tree. Theanine and tea water extract in tea were determined using GB / T 23193-2017 and GB / T8305-2013 methods, respectively, and the leaf mass was determined by weighing. Soil samples were taken from 15 cm deep soil near the rhizosphere, and 3 sampling points were taken for each tea tree. 50 g of soil was collected from each sampling point and mixed. A total of 15 tea trees were randomly sampled for analysis. The activity of urease was determined using the sodium phenol-sodium hypochlorite colorimetric method, with 1 μg of NH3-N catalyzed by 1 gram of soil filtrate representing 1 U / g. The activities of amylase and cellulase were determined by 3,5-dinitrosalicylic acid colorimetry using 1% soluble starch and 1% carboxymethyl cellulose solution as substrates, respectively. 1 U / g was defined as the amount of glucose produced per minute by catalysis of 1 μg of soil filtrate.

[0047] The measurement results of relevant indicators are shown in Table 3 below.

[0048] Table 3

[0049]

[0050] Compared with the control, the activities of urease, amylase and cellulase in the soil treated with the bacterial agent prepared by the strain of the present invention were increased by 19.0%, 18.4% and 50.0%, respectively. The theanine content of fresh leaves of spring tea of ​​tea trees increased by 11.2%, and the water extract content increased by 7.8%. The weight of fresh leaves of summer tea increased by 4.5%.

[0051] Example 4

[0052] 1. Activation of strains: 1L of liquid culture medium (10g of malt extract, 4g of yeast extract, 4g of D-glucose, 1000mL of distilled water, pH adjusted to 7.2, autoclaved), inoculated with 200mg of freeze-dried powder (1.0×10 6 CFU / g), at 25°C, with an aeration rate of 0.5 vvm and stirring at 200 rpm for 72 h to obtain the culture solution.

[0053] 2. Preparation of the microbial agent: Mix the culture solution, autoclaved tea garden soil and light calcium carbonate in a ratio of 1:50:0.5 (v / w / w), and culture for 48 hours at a temperature of 30°C and a humidity of 80% to obtain the microbial agent.

[0054] 3. Use of microbial agent: 20 days before picking spring tea, apply 400g of microbial agent to the soil 15-30cm deep under the roots of large-leaf shrub tea trees for each tea tree; 30 days before picking summer tea, apply 600g of microbial agent to the soil 15-30cm deep under the roots of tea trees for each tea tree.

[0055] 4. Index detection and results: The tea trees in the same tea garden that were treated with sterilized soil + calcium carbonate of the same mass were used as controls. Fifty fresh leaves were randomly picked from each tree. Theanine and tea water extract in tea were determined using GB / T 23193-2017 and GB / T8305-2013 methods, respectively, and the leaf mass was determined by weighing. Soil samples were taken from 15 cm deep soil near the rhizosphere, and 3 sampling points were taken for each tea tree. 50 g of soil was collected from each sampling point and mixed. A total of 15 tea trees were randomly sampled for analysis. The activity of urease was determined using the sodium phenol-sodium hypochlorite colorimetric method, with 1 μg of NH3-N catalyzed by 1 gram of soil filtrate representing 1 U / g. The activities of amylase and cellulase were determined by 3,5-dinitrosalicylic acid colorimetry using 1% soluble starch and 1% carboxymethyl cellulose solution as substrates, respectively. 1 U / g was defined as the amount of glucose produced per minute by catalysis of 1 μg of soil filtrate.

[0056] The measurement results of relevant indicators are shown in Table 4 below.

[0057] Table 4

[0058]

[0059] Compared with the control, the activities of urease, amylase and cellulase in the soil treated with the bacterial agent prepared by the strain of the present invention were increased by 33.3%, 26.3% and 85.7%, respectively. The theanine content of fresh leaves of spring tea of ​​tea trees increased by 12.9%, the water extract content increased by 15.1%, and the weight of fresh leaves of summer tea increased by 7.0%.

[0060] Although the above describes the specific implementation of the present invention in combination with the embodiments, it is not intended to limit the scope of protection of the present invention. Those skilled in the art should understand that it should be clear to those skilled in the art that these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection of the present invention.

Claims

1. A soil microorganism that promotes the growth of tea trees and improves the quality of tea leaves. The soil microorganism is taxonomically named Streptomyces cerasinus SI-3B1, deposited in Guangdong Province Microbiological Culture Collection Center, date of deposit: September 13, 2024, deposit number: GDMCC No.65129.

2. A bacterial agent comprising the soil microorganism according to claim 1.

3. The bacterial agent according to claim 2, characterized in that The bacterial agent also includes auxiliary materials.

4. The bacterial agent according to claim 2, characterized in that The auxiliary material is selected from soil or light calcium carbonate or a mixture of the two.

5. The bacterial agent according to any one of claims 2 to 4, characterized in that: The bacterial agent is prepared according to the following method: the fermentation culture solution of the soil microorganisms according to claim 1, soil and light calcium carbonate are mixed in a certain proportion, and the bacterial agent is cultured to obtain the bacterial agent.

6. The bacterial agent according to claim 5, characterized in that The bacterial agent is prepared according to the following method: The fermentation culture solution of the soil microorganisms according to claim 1, soil and light calcium carbonate are mixed in a ratio of 1-5:10-30:0.1-0.5 (v / w / w), and cultured for 48 hours at a temperature of 30° C. and a humidity of 80% to obtain a bacterial agent.

7. Use of the soil microorganisms according to claim 1 and the bacterial agents according to claims 2 to 6 in promoting the growth of tea trees and improving the quality of tea leaves.

8. The use according to claim 8, characterized in that: The application comprises: applying the soil microorganisms described in claim 1 or the bacterial agents described in claims 2-6 into the soil at the root of the tea tree at a depth of 10-40 cm.

9. The use according to claim 8 or 9, characterized in that: The method of promoting the growth of tea trees and improving the quality of tea leaves comprises: The content of theanine and water extract increased; Increased thickness of summer tea leaves; and, Increased leaf biomass.

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