阅读说明：本技术 一种短小芽孢杆菌菌肥及其应用 (Bacillus pumilus bacterial fertilizer and application thereof ) 是由 张新慧 张晓佳 郎多勇 张文晋 张瑜 崔高畅 赵建军 赵云生 于 2019-08-01 设计创作，主要内容包括：本发明公开了一种短小芽孢杆菌菌肥及其应用。本发明提供了短小芽孢杆菌(Bacillus pumilus))G5 CGMCC No.16879在增强植物的碳氮代谢中的应用。本发明短小芽孢杆菌(Bacillus pumilus)G5对于提高农产品产量、改善农产品品质、减少化肥用量、降低成本、改良土壤、保护生态环境具有重要意义。(The invention discloses a bacillus pumilus bacterial fertilizer and application thereof. The invention provides application of Bacillus pumilus (Bacillus pumilus) G5 CGMCC No.16879 in enhancing the carbon and nitrogen metabolism of plants. The Bacillus pumilus (Bacillus pumilus) G5 has important significance for improving the yield of agricultural products, improving the quality of the agricultural products, reducing the using amount of chemical fertilizers, reducing the cost, improving the soil and protecting the ecological environment.)

1. The application of the Bacillus pumilus (Bacillus pumilus) G5 or a microbial inoculum containing the Bacillus pumilus (Bacillus pumilus) G5 in any one of the following steps:

(A1) enhancing the carbon and nitrogen metabolism of plants;

(A2) preparing a product for enhancing the carbon and nitrogen metabolism of plants;

the preservation number of the Bacillus pumilus (Bacillus pumilus) G5 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 16879.

2. The application of the Bacillus pumilus (Bacillus pumilus) G5 or a microbial inoculum containing the Bacillus pumilus (Bacillus pumilus) G5 in any one of the following steps:

(B1) improving the activity of nitrate reductase in plants, or preparing products for improving the activity of nitrate reductase in plants;

(B2) increasing the activity of the sucrose invertase in the plant, or preparing a product for increasing the activity of the sucrose invertase in the plant;

the preservation number of the Bacillus pumilus (Bacillus pumilus) G5 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 16879.

3. The application of the Bacillus pumilus (Bacillus pumilus) G5 or a microbial inoculum containing the Bacillus pumilus (Bacillus pumilus) G5 in any one of the following steps:

(C1) increasing plant yield, or making a product for increasing plant yield;

(C2) improving plant quality, or preparing a product for improving plant quality;

(C3) reducing the amount of fertilizer used during plant cultivation, or preparing a product for reducing the amount of fertilizer used during plant cultivation;

(C4) improving soil, or preparing a product for improving soil;

the preservation number of the Bacillus pumilus (Bacillus pumilus) G5 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 16879.

4. The application of the Bacillus pumilus (Bacillus pumilus) G5 or a microbial inoculum containing the Bacillus pumilus (Bacillus pumilus) G5 in any one of the following steps:

(C1) promoting germination of plant seeds, or preparing a product for promoting germination of plant seeds;

(C2) improving the antioxidant capacity of the plant, or preparing a product for improving the antioxidant capacity of the plant;

(C3) increasing the content of osmoregulation substances in plants, or preparing products for increasing the content of osmoregulation substances in plants;

the preservation number of the Bacillus pumilus (Bacillus pumilus) G5 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 16879.

5. The application of the Bacillus pumilus (Bacillus pumilus) G5 or a microbial inoculum containing the Bacillus pumilus (Bacillus pumilus) G5 in any one of the following steps:

(D1) increasing the germination rate of plant seeds, or preparing a product for increasing the germination rate of plant seeds;

(D2) increasing the germination vigor of plant seeds, or preparing products for increasing the germination vigor of plant seeds;

(D3) increasing the germination index of plant seeds, or preparing a product for increasing the germination index of plant seeds;

(D4) improving the vitality index of the plant seedling, or preparing a product for improving the vitality index of the plant seedling;

(D5) increasing glutathione content in plants, or preparing products for increasing glutathione content in plants;

(D6) increasing the ascorbic acid content in plants, or preparing products for increasing the ascorbic acid content in plants;

(D7) increasing the content of soluble sugar in the plant body, or preparing a product for increasing the content of soluble sugar in the plant body;

(D8) increasing the content of soluble protein in the plant body, or preparing a product for increasing the content of soluble protein in the plant body;

the preservation number of the Bacillus pumilus (Bacillus pumilus) G5 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 16879.

6. Use according to any one of claims 1 to 5, characterized in that: the microbial inoculum is a liquid microbial inoculum;

further, the liquid microbial agent is fermentation liquor obtained by culturing the Bacillus pumilus (Bacillus pumilus)) G5 in a fermentation medium;

further, in the liquid microbial agent, the content of the Bacillus pumilus (Bacillus pumilus) G5 is 495 hundred million cfu/mL.

7. Use according to claim 6, characterized in that: the pH of the fermentation medium is 7.0, the solvent is water, and the solutes and the concentrations are as follows: maltose 4.0%, soybean peptone 3.0%, KH₂PO₄0.01 percent; the% each means g/100 mL.

8. Use according to claim 6 or 7, characterized in that: the temperature of the culture is 34 ℃, and the time is 42 h.

9. Use according to any one of claims 1 to 8, wherein: in the application, the inoculation mode is to soak plant seeds in the liquid microbial agent disclosed in claim 6.

10. The liquid microbial inoculant described in any one of claims 6 to 8.

Technical Field

The invention relates to the field of microorganisms and agriculture, in particular to a bacillus pumilus bacterial fertilizer and application thereof.

Background

Along with the development of society, people pay more and more attention to environmental pollution, food safety and human health; and the use of a large amount of chemical fertilizers and pesticides in agricultural production not only causes environmental pollution and quality reduction of agricultural products, but also causes the overproof of harmful substances such as pesticides and heavy metals and seriously harms human health. The agricultural department in 2015 provided the goal of realizing zero increase of the use amount of chemical fertilizers and pesticides in 2020, greatly promoted the reduction and the increase of the use amount of chemical fertilizers and pesticides, and led the development mode of the planting industry to change, wherein the popularization and the application of microbial agents are important components for realizing the goal, so that on one hand, agricultural resources can be fully utilized, and on the other hand, the wide use of agricultural microorganisms can greatly reduce the use amount of chemical pesticides and chemical fertilizers, reduce environmental pollution, reduce production cost, improve the quality and the benefit of agricultural products, increase the market competitiveness of agricultural products, and provide new biological yield and resources for the adjustment and the earning of the agricultural industrial structure.

In many areas, severe drought and saline-alkali stress exists, and high yield and quality of plants are difficult to obtain under the stress, so that a method for ensuring the quality and yield of the plants and being eco-friendly is urgently sought. The microbial fertilizer is a novel fertilizer biological product which leads crops to obtain required nutrients by the life activities of active microorganisms. The microbial agent is a low-carbon, purely natural, non-toxic, harmless and pollution-free organic microbial agent, and has the functions of improving soil fertility, increasing the number and activity of beneficial microorganisms in soil, improving soil activation characters, preventing soil hardening, improving soil fertility preservation and water retention capacity and the like, promoting crop growth, improving crop yield, improving and reducing agricultural product quality and the like.

Disclosure of Invention

The invention aims to provide a bacillus pumilus bacterial fertilizer and application thereof.

In a first aspect, the present invention claims the use of Bacillus pumilus (Bacillus pumilus) G5 or a bacterial agent comprising said Bacillus pumilus (Bacillus pumilus) G5 in any of:

(A1) enhancing the carbon and nitrogen metabolism of plants;

(A2) preparing a product for enhancing the carbon and nitrogen metabolism of plants;

the preservation number of the Bacillus pumilus (Bacillus pumilus) G5 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 16879.

In a second aspect, the present invention claims the use of Bacillus pumilus (Bacillus pumilus) G5 or a bacterial agent comprising said Bacillus pumilus (Bacillus pumilus) G5 in any of:

(B1) increasing Nitrate Reductase (NR) enzyme activity in plants, or preparing products for increasing nitrate reductase enzyme activity in plants;

(B2) increasing the activity of sucrose Invertase (INV) in plants, or preparing products for increasing the activity of sucrose invertase in plants;

the preservation number of the Bacillus pumilus (Bacillus pumilus) G5 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 16879.

In a third aspect, the invention claims the application of Bacillus pumilus (Bacillus pumilus) G5 or a microbial inoculum containing the Bacillus pumilus (Bacillus pumilus) G5 in any one of the following:

(C1) increasing plant yield, or making a product for increasing plant yield;

(C2) improving plant quality, or preparing a product for improving plant quality;

(C3) reducing the amount of fertilizer used during plant cultivation, or preparing a product for reducing the amount of fertilizer used during plant cultivation;

(C4) improving soil, or preparing a product for improving soil;

the preservation number of the Bacillus pumilus (Bacillus pumilus) G5 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 16879.

In a fourth aspect, the present invention claims the use of Bacillus pumilus (Bacillus pumilus) G5 or a bacterial agent comprising said Bacillus pumilus (Bacillus pumilus) G5 in any one of the following:

(C1) promoting germination of plant seeds, or preparing a product for promoting germination of plant seeds;

(C2) improving the antioxidant capacity of the plant, or preparing a product for improving the antioxidant capacity of the plant;

(C3) increasing the content of osmoregulation substances in plants, or preparing products for increasing the content of osmoregulation substances in plants;

the preservation number of the Bacillus pumilus (Bacillus pumilus) G5 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 16879.

In a fifth aspect, the present invention claims the use of Bacillus pumilus (Bacillus pumilus) G5 or a bacterial agent comprising said Bacillus pumilus (Bacillus pumilus) G5 in any one of the following:

(D1) increasing the germination rate of plant seeds, or preparing a product for increasing the germination rate of plant seeds;

(D2) increasing the germination vigor of plant seeds, or preparing products for increasing the germination vigor of plant seeds;

(D3) increasing the germination index of plant seeds, or preparing a product for increasing the germination index of plant seeds;

(D4) improving the vitality index of the plant seedling, or preparing a product for improving the vitality index of the plant seedling;

(D5) increasing glutathione content in plants, or preparing products for increasing glutathione content in plants;

(D6) increasing the ascorbic acid content in plants, or preparing products for increasing the ascorbic acid content in plants;

(D7) increasing the content of soluble sugar in the plant body, or preparing a product for increasing the content of soluble sugar in the plant body;

(D8) increasing the content of soluble protein in the plant body, or preparing a product for increasing the content of soluble protein in the plant body;

the preservation number of the Bacillus pumilus (Bacillus pumilus) G5 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 16879.

In the aspects, the active ingredient of the microbial inoculum is the Bacillus pumilus (Bacillus pumilus) G5, and other auxiliary materials, auxiliary agents and the like can be contained.

Further, in the invention, the microbial inoculum is a liquid microbial inoculum.

Further, the liquid microbial agent is a fermentation liquid obtained by culturing the Bacillus pumilus (Bacillus pumilus)) G5 in a fermentation medium.

Specifically, in the liquid microbial agent, the content of the Bacillus pumilus (Bacillus pumilus) G5 is 495 hundred million cfu/mL.

In a specific embodiment of the present invention, the pH of the fermentation medium is 7.0, the solvent is water, and the solutes and concentrations are as follows: maltose 4.0%, soybean peptone 3.0%, KH₂PO₄0.01 percent; the% each means g/100 mL. The temperature of the culture is 34 ℃, and the time is 42 h. The rotating speed of the shaking table in the culture process is 230 r/min.

Germination rate (%) × 100% (number of germinated seeds/number of test seeds);

germination vigor (%) × (number of germinated seeds/number of test seeds in a prescribed date) × 100%;

germination index ∑ (germination number on day of the germination test/day of the germination test).

Seedling vigor index is the seedling growth (length or weight) versus germination index.

In each of the above applications, the inoculation is by immersing the plant seed in the liquid microbial inoculum as described in the second aspect above. Further, the plant seeds are soaked in 100-fold dilution of the liquid microbial agent with the content of 495 hundred million cfu/mL of the Bacillus pumilus (Bacillus pumilus) G5 for 6-8 hours.

Still further, the step of surface sterilizing the plant seed may be included prior to immersing the plant seed in the liquid microbial inoculant.

In the present invention, the plant may be a dicotyledonous plant or a monocotyledonous plant. Further, the dicotyledonous plant may be a glycyrrhiza plant. Further, the glycyrrhiza plant may be a leguminous plant. In a particular embodiment of the invention, the plant is in particular licorice, a plant of the family leguminosae.

In the use according to the fourth to sixth aspects, under normal conditions, salt stress conditions or drought stress conditions, the enhancing of carbon and nitrogen metabolism of a plant, the enhancing of Nitrate Reductase (NR) enzyme activity in a plant, the enhancing of sucrose Invertase (INV) enzyme activity in a plant, the promoting of germination of plant seeds (enhancing of seed vigor), the enhancing of oxidation resistance of a plant, the enhancing of content of osmoregulation substances in a plant, the enhancing of germination rate of plant seeds, the enhancing of germination vigor of plant seeds, the enhancing of germination index of plant seeds, the enhancing of vigor index of plant seedlings, the enhancing of Glutathione (GSH) content in a plant, the enhancing of ascorbic acid (AsA) content in a plant, the enhancing of soluble sugar content in a plant and/or the enhancing of soluble protein content in a plant.

Further, the salt stress is 150mmol/L NaCl; the drought stress was 10% PEG.

Experiments prove that the Bacillus pumilus G5 provided by the invention can enhance plant carbon and nitrogen metabolism, promote plant seed germination, improve the oxidation resistance of plants and improve the content of osmotic adjusting substances in plants, and the microbial fertilizer with plant stress resistance is developed by scientifically utilizing microbial resources and considering actual production and market demands. The invention has important significance for improving the yield of agricultural products, improving the quality of the agricultural products, reducing the using amount of chemical fertilizers, reducing the cost, improving the soil and protecting the ecological environment.

Deposit description

The strain name: bacillus pumilus

Latin name: bacillus pumilus

According to the biological materials (strains): g5

The preservation organization: china general microbiological culture Collection center

The preservation organization is abbreviated as: CGMCC (China general microbiological culture Collection center)

Address: xilu No.1 Hospital No. 3 of Beijing market facing Yang district

The preservation date is as follows: 12 month and 6 days 2018

Registration number of the preservation center: CGMCC No.16879

Drawings

FIG. 1 shows the results of the radicle length, radicle thickness, germ length, and germ thickness of the licorice seedlings among different treatment groups.

FIG. 2 shows the germination rates of seedlings of Glycyrrhiza uralensis between different treatment groups.

FIG. 3 shows the germination potential of licorice seedlings among different treatment groups.

FIG. 4 shows the germination index of licorice seedlings among different treatment groups.

FIG. 5 shows the vitality index of licorice seedlings among different treatment groups.

FIG. 6 shows the change of the antioxidant system of Glycyrrhiza uralensis between different treatment groups.

FIG. 7 shows the enzyme activities of NR and INV in the carbonitriding process of licorice among different treatment groups.

FIG. 8 shows the change of the permeation-regulating substances of licorice among different treatment groups.

In the figures, the representation differs significantly between different lower case letters.

Detailed Description

The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.

Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

The Bacillus pumilus (Bacillus pumilus) G5 is an endophyte separated from collected Glycyrrhiza uralensis, and the strain with the highest homology is CTSP17 through morphological, physiological and biochemical characteristic identification and comparison of a 16SrDNA ribosome database; EU855198, CTSP 18; EU855199, CTSP 19; EU855200, CTSP 20; EU855201 finally identifies the strain G5 as Bacillus pumilus (Bacillus pumilus). The strain is preserved in China general microbiological culture Collection center in 2018, 12 months and 06 months, and the preservation number is CGMCC No. 16879.