Streptomyces F11 as well as microbial preparation and application thereof

Abstract

The invention relates to streptomyces F11 as well as a microbial preparation and application thereof, and aims to solve the technical problems that the utilization rate of nitrogen resources in current soil is low and environmental pollution is easily caused. Streptomyces sp. F11 is screened from a soil microbial community to which a silicon fertilizer is applied, and the preservation number is CGMCC No.33657. The Streptomyces sp. F11 not only has a nitrogen fixation effect, but also can improve available nitrogen in soil so as to promote the growth and development of rice; and by combining / multiplexing with silicon, the rice panicle number or / and dry weight can be increased. According to the present invention, the dependence on the traditional nitrogen fertilizer can be reduced, the nitrogen resource in the soil can be efficiently utilized at the low cost, the rice growth can be promoted, the rice yield can be increased, and the development of the green ecological agriculture can be promoted.

Description

Technical Field

[0001] The present invention relates to the technical field of bacterial fertilizers, and particularly relates to a Streptomyces F11, its microbial preparation and application. Background Art

[0002] Rice is an important food crop in China, and its growth depends on the key nutrient element of nitrogen. However, the nitrogen resources available for plants to absorb in the soil are limited and difficult to meet the needs of high-yield crops. Therefore, applying nitrogen fertilizer has become a necessary measure to supplement soil nitrogen and maintain the nitrogen balance in farmland. However, with the increasing annual use of nitrogen fertilizer in China, its impact on the environment has become increasingly prominent. In order to achieve the sustainable development of agriculture and reduce the dependence on nitrogen fertilizer, it is particularly important to rationally utilize resources.

[0003] As a new type of fertilizer, silicon fertilizer can not only enhance the resistance of rice to biotic and abiotic stresses, promote its growth and development and increase yield, but also increase the nitrogen content in the soil by affecting the soil microbial community structure, thereby promoting the growth of rice. Therefore, exploring new microorganisms that can improve the soil nitrogen use efficiency under the action of silicon fertilizer is of great significance for developing soil nitrogen resources and developing green ecological agriculture. It can not only improve the utilization efficiency of soil nitrogen, but also reduce the use of chemical fertilizers and reduce environmental pollution, which is an effective way to achieve the sustainable development of agriculture.

[0004] The information disclosed in this background art section is only for enhancing the understanding of the background art of the present disclosure, and should not be regarded as an admission or any form of implication that this information constitutes the prior art known to those skilled in the art. Summary of the Invention

[0005] While silicon fertilizer promotes the growth and development of rice and increases yield, it can also increase the nitrogen content in the soil by regulating the soil microbial community structure; based on this, the inventors of the present application screened a Streptomyces F11 that can significantly increase the ammonium nitrogen and nitrate nitrogen contents in the soil of rice roots under the action of silicon fertilizer, thereby promoting the growth of rice. While reducing the dependence on traditional nitrogen fertilizers, it can efficiently utilize the nitrogen resources in the soil at a low cost, promote the growth of rice and increase yield, thereby promoting the development of green ecological agriculture.

[0006] According to one aspect of the present disclosure, there is provided a Streptomyces ( Streptomyces sp. ) F11, which was deposited at the China General Microbiological Culture Collection Center (Address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences) on February 26, 2024, and its deposit number is CGMCC No. 33657.

[0007] According to a second aspect of the present disclosure, a microbial preparation is provided, which contains the Streptomyces F11 or / and its metabolite products.

[0008] According to a third aspect of the present disclosure, the Streptomyces F11 or the microbial preparation is applied in any one of the following (1) to (6): (1) Nitrogen fixation or preparation of nitrogen fixation products; (2) Promoting plant growth and development or preparation of products for promoting plant growth and development; (3) Increasing the content of available nitrogen in the soil or preparation of products for increasing the content of available nitrogen in the soil; (4) Increasing the plant height or preparation of products for increasing the plant height; (5) Combining / using in combination with silicon to increase the number of rice panicles and / or dry weight, or preparation of products for combining / using in combination with silicon to increase the number of rice panicles and / or dry weight; (6) Combining / using in combination with silicon to promote plant growth and development, or preparation of products for combining / using in combination with silicon to promote plant growth and development.

[0009] In some embodiments of the present disclosure, the available nitrogen includes at least one of ammonium nitrogen and nitrate nitrogen.

[0010] In some embodiments of the present disclosure, the plant includes rice.

[0011] According to a fourth aspect of the present disclosure, a product with an active ingredient being any one of the following A to C is provided: A. The Streptomyces F11; B. The metabolite of the Streptomyces F11; C. At least two of the Streptomyces F11, its metabolite products, and silicon; This product has at least one of the following functions or effects (1) to (3): (1) Nitrogen fixation; (2) Promoting plant growth and development; (3) Increasing the number of rice panicles and / or dry weight.

[0012] One or more technical solutions provided in the embodiments of the present application have at least any one of the following technical effects or advantages: A Streptomyces F11 was screened from the Streptomyces community in silicon-added soil. It has nitrogen fixation function and can significantly increase the content of available nitrogen in the soil (such as ammonium nitrogen and nitrate nitrogen), thereby promoting the growth and development of rice; and the Streptomyces F11 is combined / used in combination with silicon to increase the number of rice panicles and / or dry weight. While reducing the dependence on traditional nitrogen fertilizers, it efficiently utilizes the nitrogen resource in the soil at a low cost, promotes the growth of rice, increases the yield, and thus promotes the development of green ecological agriculture. Brief Description of the Drawings

[0013] Figure 1 This shows the changes in the number of rice panicles and dry weight during different treatments in one embodiment of this application.

[0014] Figure 2 This is the microbial network diagram for different treatments in one embodiment of this application.

[0015] Figure 3 This shows the changes in soil nitrogen during different treatments in one embodiment of this application.

[0016] Figure 4 This shows the changes in the abundance of Streptomyces during different treatments in one embodiment of this application.

[0017] Figure 5 This is the colony map of Streptomyces on a nitrogen-free medium in one embodiment of this application.

[0018] Figure 6 This shows the effects of Streptomyces F11 on soil ammonium nitrogen and nitrate nitrogen in one embodiment of this application.

[0019] Figure 7 This shows the effects of Streptomyces F11 on the plant height and chlorophyll of rice in one embodiment of this application.

[0020] In the above figures, different lowercase letters represent significant differences between the two, p < 0.05. Detailed Embodiments

[0021] To better understand the technical solution of the present invention, the above technical solution will be described in detail below in conjunction with the accompanying drawings of the specification and specific embodiments.

[0022] In the following embodiments, the instruments and equipment involved are all conventional instruments and equipment unless otherwise specified; the reagents involved are all commercially available conventional reagents unless otherwise specified; the test methods and detection methods involved are all conventional methods unless otherwise specified.

[0023] Example 1: Isolation, Identification and Preservation of Streptomyces F11 1. Isolation of Streptomyces F11 The pot experiment was carried out at the Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences from July 29 to September 26, 2023. A rice pot experiment with soil from Shaoxing City, Zhejiang Province (SX) and Gaoyou City, Jiangsu Province (GY) in China was conducted with treatments of no silicon addition (Si0) and silicon addition (Si1, adding 0.25 g of Na2SiO3 per kg of soil). The results showed that (as Figures 1 to 4As shown in the figure, silicon addition treatment can significantly increase the panicle number and dry weight of rice, change the microbial community structure, and at the same time significantly increase the ammonium nitrogen and nitrate nitrogen contents and the abundance of Streptomyces in the soil. Zeng Xian isolated a Streptomyces from the rhizosphere soil of rice pot experiments with silicon addition treatment through methods such as primary screening by gradient dilution plate method, and named it Streptomyces F11.

[0024] The specific method of the gradient dilution plate method is as follows: Grind the collected samples through a 200-mesh sieve to prepare 7 dilutions of bacterial suspensions with concentrations of 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 . Uniformly coat them on 1 / 2 R2A medium, culture them in a constant temperature incubator at 30°C for 2 - 3 days, pick different morphological single colonies and streak them on LB medium 2 - 3 times until pure culture is obtained to obtain single colonies.

[0025] 1 / 2 R2A medium: 0.125 g of yeast extract, 0.125 g of tryptone, 0.125 g of casein hydrolysate, 0.125 g of glucose, 0.125 g of soluble starch, 0.075 g of sodium pyruvate, 0.075 g of K2HPO4, 0.0125 g of MgSO4·7H2O, 15 g of agar, 1 L of deionized water, pH 7 - 7.4.

[0026] 2. Identification of Streptomyces F11 Culture the screened strains on LB solid medium (10 g of tryptone, 10 g of sodium chloride, 5 g of yeast extract, 15 g of agar, 1 L of deionized water, pH 7.0), observe their colony characteristics and physiological and biochemical characteristics such as color with reference to the Manual of Systematic Identification of Common Bacteria; use the universal bacterial primers GM3F and GM4R for PCR amplification to obtain a 1200-bp product, sequence it, and perform sequence alignment on the sequencing results through Blast.

[0027] 3. Preservation of Streptomyces F11 The screened and identified Streptomyces F11 was preserved at the General Microbiology Center of the China Committee for Culture Collection of Microorganisms on February 26, 2024. Address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The taxonomic name is Streptomyces Streptomyces sp . F11, and the preservation number is CGMCC No. 33657.

[0028] Example 2. Verification of the nitrogen fixation function of Streptomyces F11 In this example, Streptomyces ([[]] Streptomyces sp [[]].) F11 was inoculated into Ashby nitrogen-free medium and cultured at 30 °C for 7 days to observe the growth status of the strain. The results showed (as shown in [[]] Figure 5 [[]]) that Streptomyces ([[]] Streptomyces sp [[]].) F11 could grow well in nitrogen-free medium, indicating its ability of free-living nitrogen fixation.

[0029] The raw material formula of the above Ashby nitrogen-free medium (nitrogen-fixing bacteria) is as follows: KH2PO4 0.2 g, MgSO4·7H2O 0.2 g, NaCl 0.2 g, CaCO3 5.0 g / L, mannitol 10.0 g, CaSO4 0.1 g, agar 15.0 g, deionized water 1 L, pH 6.9 - 7.1.

[0030] Example 3. Effects of Streptomyces F11 on soil ammonium nitrogen and nitrate nitrogen Soil samples were collected from the soil surface layer (0 to 20 cm) in Shaoxing City (SX), Zhejiang Province, China and Gaoyou City (GY), Jiangsu Province. The soil was sieved through a 2 mm sieve to remove stones and roots, and then dried in the air and thoroughly mixed. 10 g of soil was weighed into a 50 mL serum bottle, and sterile water was added to 2 cm above the soil surface. The bottle was sealed with a sealing film and sterilized by high-pressure steam for 30 min. 5 ml of bacterial solution (F11) was inoculated into the soil of the experimental group, and 5 ml of sterile water (CK) was added to the control treatment. The samples were cultured in the dark at 25 °C, and each treatment had 4 replicates. Soil samples were destructively collected on the 16th day of culture to detect the concentrations of soil nitrate nitrogen and ammonium nitrogen. The results are as shown in [[]] Figure 6 [[]], indicating that Streptomyces significantly increased the ammonium nitrogen and nitrate nitrogen in the soil compared with the control group.

[0031] Example 4. Effects of Streptomyces F11 on rice growth Rice seeds were soaked in 8% dilute nitric acid solution for sterilization for 12 h, then thoroughly rinsed with distilled water, and then soaked in distilled water in the dark at 30 °C for 3 d for germination. After germination, the seeds were sown in a seedling tray containing 80 g of sterilized soil. Two treatments were set: CK (without adding bacterial solution) and F11 (adding Streptomyces suspension). The bacterial suspension was added immediately after sowing. The plants were placed in a growth chamber with a daytime room temperature of 32 °C, 16 h of light, a nighttime room temperature of 28 °C, and 8 h of darkness. The plant height and chlorophyll were measured after 8 d and 12 d of growth. The results are as shown in [[]] Figure 7 [[]], indicating that Streptomyces F11 significantly increased the plant height and chlorophyll of rice compared with the control.

[0032] ​​​​​​​​​​Although some preferred embodiments of the present invention have been described, additional changes and modifications can be made to these embodiments by those skilled in the art once they learn the basic creative concept. Therefore, the appended claims of this application are intended to be interpreted to include the preferred embodiments as well as all changes and modifications that fall within the scope of this application.

[0033] Obviously, those skilled in the art can make various changes and modifications to the present disclosure without departing from the spirit and scope of its inventive concept. Thus, if these modifications and variations to the present disclosure fall within the scope of the claims of this application and their equivalent technologies, this application is also intended to include these modifications and variations.

Claims

1. A Streptomyces strain ( Streptomyces sp. ) F11, with the deposit number of CGMCC No. 33657.

2. A microbial preparation containing the Streptomyces sp. F11 described in claim 1 and / or its metabolite products.

3. The application of the Streptomyces sp. F11 described in claim 1 or the microbial preparation described in claim 2 in any one of the following (1) to (6): (1) Nitrogen fixation or preparation of nitrogen fixation products; (2) Promoting plant growth and development or preparation of products for promoting plant growth and development; (3) Increasing the content of available nitrogen in the soil or preparation of products for increasing the content of available nitrogen in the soil; (4) Increasing plant height or preparation of products for increasing plant height; (5) Combining / using in combination with silicon to increase the number of rice panicles and / or dry weight, or preparation of products for combining / using in combination with silicon to increase the number of rice panicles and / or dry weight; (6) Combining / using in combination with silicon to promote plant growth and development, or preparation of products for combining / using in combination with silicon to promote plant growth and development.

4. The application according to claim 3, wherein, The available nitrogen includes at least one of ammonium nitrogen and nitrate nitrogen.

5. The application according to claim 3, characterized in that, The plants include rice.

6. A product with an active ingredient being any one of the following A to C: A. The Streptomyces sp. F11 described in claim 1; B. The metabolite of the Streptomyces sp. F11 described in claim 1; C. At least two of the Streptomyces sp. F11 described in claim 1, its metabolite products, and silicon; This product has at least one of the following functions or effects (1) to (3): (1) Nitrogen fixation; (2) Promoting plant growth and development; (3) Increasing the number of rice panicles and / or dry weight.

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