CN115261267B - Application of brown alginate oligosaccharides in promoting bacillus movement and plant rhizosphere colonisation - Google Patents
Application of brown alginate oligosaccharides in promoting bacillus movement and plant rhizosphere colonisation Download PDFInfo
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- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
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- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
Abstract
The invention provides an application of brown algae oligosaccharide in promoting bacillus movement and bacillus in plant rhizosphere field planting, and belongs to the technical field of microorganisms. The bacillus comprises one or more of bacillus subtilis, bacillus belicus and bacillus amyloliquefaciens, and the brown alginate oligosaccharides remarkably promote the movement capability of the bacillus. The invention also provides a bacillus microbial inoculum, which comprises bacillus and brown alginate oligosaccharides, wherein the brown alginate oligosaccharides can obviously promote the movement capability of the bacillus, so that the bacillus can be quickly and effectively planted in the rhizosphere of crops, the microbial inoculum prepared by combining the brown alginate oligosaccharides and the bacillus can obviously promote the growth of plants, and an effective way is provided for reducing the application of chemical pesticide fertilizers and promoting the sustainable development of agriculture.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to application of brown alginate oligosaccharides in promoting the movement capacity of bacillus.
Background
In agricultural production, plant rhizosphere growth promoting bacteria play an important role in crop health and high yield, and the application of the plant rhizosphere growth promoting bacteria is an effective way for realizing the reduction of application of chemical pesticide fertilizers in China and promoting sustainable development of agriculture. At present, scholars at home and abroad have separated from rhizosphere of important crops such as rice, wheat, corn, cotton and the like to obtain a large number of biocontrol strains with disease prevention and growth promotion effects, and part of strains are used for commercial production. However, after the biocontrol bacteria enter the soil, the biocontrol bacteria face competition of indigenous microorganisms and complex soil environment conditions, dominant populations are difficult to form, the effective biocontrol function is exerted, and the problems of large indoor and field control effect difference, unstable field control effect and the like often exist in practical application, so that the agricultural application of the biocontrol bacteria is hindered. Efficient rhizosphere colonization is an important precondition for microorganisms to exert various plant probiotic functions, so that the problem of difficult colonization of plant rhizosphere by microorganism strains is a key for breaking the limitation of biocontrol bacterium agricultural application, and the colonization of plant rhizosphere by the microorganism strains is obviously influenced by the movement capability of the microorganism strains on soil or plant surfaces.
The brown alginate oligosaccharides are a compound with strong solubility, easy absorption, safety and no toxicity, and the research finds that the brown alginate oligosaccharides have the functions of oxidation resistance, antibacterial activity and treatment of human diseases. At present, the brown alginate oligosaccharides have wide development prospects in the fields of medicine development and functional food development, but no report on the ability of utilizing the brown alginate oligosaccharides to promote bacterial movement exists.
Disclosure of Invention
In view of the above, the invention aims to provide the application of the brown alginate oligosaccharides in improving the movement capability of bacillus and promoting the bacillus to colonize the plant rhizosphere, and the brown alginate oligosaccharides are utilized to effectively improve the movement capability of the bacillus on the soil or the plant surface so as to promote the colonization of the strain on the plant rhizosphere, and meanwhile, the combined application of the brown alginate oligosaccharides and the bacillus can also effectively promote the growth of plants.
In order to achieve the aim of the invention, the invention provides the application of the brown alginate oligosaccharides in promoting the movement of bacillus.
Another object of the present invention is to provide the use of brown alginate oligosaccharides for promoting the colonization of plants by Bacillus.
Optionally, the bacillus includes: one or more of bacillus subtilis, bacillus belicus and bacillus amyloliquefaciens.
Optionally, the preparation method of the brown alginate oligosaccharides comprises the following steps: dissolving sodium alginate in water, adding enzyme for cleavage, centrifuging, and lyophilizing supernatant to obtain brown alginate oligosaccharide.
Preferably, in the step of preparing the brown alginate oligosaccharides, the enzyme is alginic acid lyase, the enzymolysis temperature is 25-32 ℃, and the enzymolysis time is 12-18h.
Optionally, the molecular weight of the brown alginate oligosaccharides is 1000-4000D.
The invention also aims to provide a bacillus microbial inoculum, which comprises brown alginate oligosaccharides and bacillus.
Optionally, the brown alginate oligosaccharides are used at a concentration of 1.0-7.0g/L.
Optionally, the dosage form of the microbial inoculum can be water aqua, wettable powder, water dispersible granules or water suspending agent.
The invention also aims to provide the application of the bacillus agent in promoting plant growth.
Compared with the prior art, the invention has the following beneficial effects:
the brown alginate oligosaccharide has low molecular weight, strong water solubility, high stability, safety and no toxicity. The method not only can provide a carbon source for the growth of bacillus, but also can effectively improve the movement capability of the bacillus, and in a certain amount of brown alginate oligosaccharides addition range, the promotion effect on the movement capability of the strain is positively correlated with the concentration of the brown alginate oligosaccharides; the invention adds the brown alginate oligosaccharides into bacillus bacterial agent, and provides new application to brown alginate oligosaccharides.
The bacillus microbial inoculum provided by the invention comprises the brown alginate oligosaccharides and the bacillus, wherein the brown alginate oligosaccharides in the microbial inoculum can obviously promote the movement capacity of the bacillus, and further can promote the effective colonization of bacillus strains in the rhizosphere of crops, thereby achieving the purpose of further improving the biocontrol microbial inoculum product. The bacillus added in the bacillus microbial inoculum provided by the invention is one of soil and plant microecological dominant microorganism populations, and can play a role in the modes of nutrition competition, plant growth promotion, plant resistance induction and antibacterial substance secretion, and the brown algae oligosaccharide and the bacillus are applied in a combined way, so that the movement capacity of the strain on the soil or the plant surface is promoted, the bacillus can be quickly and effectively colonized on the plant rhizosphere to form dominant strains, and further the growth of plants can be obviously promoted.
Description of biological preservation
Bacillus bailii EM-1 (Latin name Bacillus velezensis EM-1) of the invention, preservation unit: china general microbiological culture Collection center, preservation address: no. 3, no.1, no. 3, deposit number, north chen west road, korea, beijing city, korea: cgmccno.21131, date of preservation: 11/09 in 2020.
Drawings
Fig. 1: the influence of sugar from different sources on promoting the mobility of bacillus amyloliquefaciens Cas02, wherein CK is blank, COS is chitosan oligosaccharide, HSG is brown alginate oligosaccharide, HT is enteromorpha polysaccharide, KLG is carrageenan oligosaccharide, KLJ is carrageenan, SA is sodium alginate, YZ is fucoidin, and the sugar addition concentration is 5.0g/L;
fig. 2: the influence of the brown alginate oligosaccharides with different concentrations on the motility of bacillus amyloliquefaciens Cas02 is that the addition concentration is 1.0g/L,2.5g/L,5.0g/L and 10.0g/L respectively;
fig. 3: the brown algae oligosaccharide promotes effective colonization of bacillus amyloliquefaciens Cas02 in plant rhizosphere, wherein CK is applied sterile water, cas02 is applied only bacillus amyloliquefaciens Cas02 bacterial liquid, and HSG+Cas02 is applied bacillus amyloliquefaciens Cas02 bacterial liquid containing brown algae oligosaccharide;
fig. 4: agronomic traits of tobacco seedling growth are treated differently, wherein CK is irrigating sterile water, HSG is irrigating brown algae oligosaccharide solution, C is irrigating bacillus amyloliquefaciens Cas02 bacterial liquid, and HSG+C is combined application of brown algae oligosaccharide and bacillus amyloliquefaciens.
Detailed Description
The invention provides an application of brown alginate oligosaccharides in improving the movement capacity of bacillus and promoting the colonization of plant rhizosphere. The brown algae oligosaccharide provided by the invention can provide a carbon source for the growth of bacillus, and can obviously improve the movement capability of bacillus, so that the effective colonization of bacillus in plant rhizosphere is obviously promoted; the bacillus in the invention is rhizosphere growth-promoting bacteria, can play a role in nutrition competition, plant growth promotion, plant resistance induction and antibacterial substance secretion, and has biological activities of phosphate solution, potassium solution and nitrogen fixation.
The bacillus of the invention comprises one or more of bacillus subtilis, bacillus bailii and bacillus amyloliquefaciens. The bacillus is preferably bacillus amyloliquefaciens, more preferably bacillus amyloliquefaciens Cas02, which is a strain disclosed in Chinese patent CN108624543A and is preserved in China general microbiological culture Collection center (address: north Xiyun No.1, soy of Beijing, korea, china) of China general microbiological culture Collection center (China, no. 3) of China, 3 months, 26 days, and the biological preservation number is CGMCC No.15514. The bacillus subtilis is preferably bacillus subtilis Tpb55 disclosed in a document Biocontrol potential of antagonist Bacillus subtilis Tpb and 55 against tobacco black shank (Han, T.et al.BioControl,2016,61 (2), 195-205.) and is stored in the China general microbiological culture Collection center (address: north Chen West Lu 1, G.Yang area, beijing, china) at a date of 29 in 2008, and the biological preservation number is CGMCC No.2853. The bacillus belgium in the invention is preferably bacillus belgium EM-1, and the strain is preserved in China general microbiological culture Collection center (address: north Xielu No.1, 3 in the Korean region of Beijing, china) on the 11 th month 09 of 2020, and the biological preservation number is CGMCC No.21131. The bacillus added in the invention has strong stress resistance, can promote plant growth and inhibit the invasion of pathogenic bacteria to plants.
The brown alginate oligosaccharides in the invention can effectively promote the movement ability of bacillus, and the promotion effect on the movement ability of bacillus in the brown alginate oligosaccharide adding range is positively correlated with the concentration of brown alginate oligosaccharides; and the brown algae oligosaccharide can obviously promote the effective colonization of bacillus strains in the rhizosphere of crops.
As an embodiment, the method for preparing the brown alginate oligosaccharides of the present invention comprises: dissolving sodium alginate in water, adding enzyme for cleavage, centrifuging, and vacuum freeze drying supernatant to obtain brown alginate oligosaccharide.
According to the invention, water is added into sodium alginate according to the feed liquid ratio of 2-6:100g/mL, and the feed liquid ratio is preferably 4-5.5:100g/mL; the present invention is not particularly limited to the water to be added, and may be any water conventionally used in the art for testing, and distilled water may be added as an embodiment.
The enzyme added in the invention is alginic acid lyase, and the addition concentration of the alginic acid lyase is 0.1-2.5g/L, preferably 0.5-1.5g/L; the enzymolysis temperature is 25-32deg.C, preferably 28-30deg.C; the enzymolysis time is 12-18h, preferably 15-16h; centrifuging the enzymolysis liquid to remove undegraded polysaccharide, wherein the centrifugation parameters are as follows: 5000-10000rpm,3-10min, preferably 6000-8000rpm,5-8min.
The brown algae oligosaccharide solution obtained by centrifugation is dried, the drying mode of the brown algae oligosaccharide is not particularly limited, the conventional drying mode in the field is adopted, and as an implementation mode, the vacuum freeze drying mode is adopted, and the drying time is 24-72 hours, preferably 36-48 hours;
the molecular weight of the brown alginate oligosaccharides added in the invention is preferably 1000-4000D, and researches show that the brown alginate oligosaccharides have different effects on promoting plant growth when the molecular weights of the brown alginate oligosaccharides are different, and the invention researches show that the brown alginate oligosaccharides with the molecular weights of 1000-4000D can effectively improve the bioactivity of the brown alginate oligosaccharides, are beneficial to the absorption and utilization of organisms and improve the movement capability of bacillus.
The invention also provides a bacillus microbial inoculum, which comprises brown alginate oligosaccharides and bacillus.
The method for culturing bacillus of the present invention is not particularly limited, and as one embodiment, the method for culturing bacillus of the present invention includes: the strain is placed in LB liquid medium for culture, and the culture condition is 28 ℃ for aerobic culture for 12 hours.
Wherein the LB culture medium is a conventional culture medium in the field, and the components (g/L) are as follows: tryptone 10.0, yeast extract 5.0, sodium chloride 10.0, pH 7.0.+ -. 0.2; meanwhile, as an implementation way, the liquid culture medium after the strain inoculation is placed on a shaking table for shaking culture at 120-160rpm, preferably 130-150 rpm.
In the present invention, the bacterial liquid obtained by the culture was centrifuged at 600 rpm for 5min to obtain bacterial cells, and the bacterial cells were resuspended to OD with sterile water 600 From 0.1 to 1.0, preferably by adjusting the OD 600 0.2-0.5; the brown alginate oligosaccharides are added into bacillus bacterial liquid, and the concentration of brown alginate oligosaccharides in the bacterial liquid is preferably 1.0-7.0g/L, and more preferably 1.5-5.0g/L.
The preparation formulation of the bacillus bacteria agent can be water aqua, wettable powder, water dispersible granules or water suspending agent.
The bacillus bacteria agent also comprises an additive, wherein the additive comprises one or more of a dispersing agent, a wetting agent, a disintegrating agent, a binding agent, an antifoaming agent, an anti-freezing agent, a thickening agent, a filler and a solvent.
Wherein the dispersing agent is an anionic dispersing agent and/or a nonionic dispersing agent, and one or more of sodium lignin sulfonate, sodium naphthalene sulfonate formaldehyde condensate, sodium methylene dinaphthyl sulfonate, formaldehyde condensate sulfate, polycarboxylate, alkylphenol polyoxyethylene phosphate and fatty acid polyoxyethylene ester can be selected;
the wetting agent can be one or more selected from sodium dodecyl sulfate, sodium dodecyl benzene sulfonate, fructus Gleditsiae Abnormalis powder, fructus Sapindi Mukouossi powder, tea seed cake powder and radix et rhizoma Rhei powder BX;
the disintegrating agent can be one or more selected from bentonite, ammonium sulfate, aluminum chloride, urea, magnesium chloride and glucose;
the binder may be selected from one or more of starch, diatomaceous earth, cyclodextrin, rosin, carboxymethyl cellulose, carboxyethyl cellulose and carboxymethyl cellulose salt;
the defoamer can be one or more selected from C8-C20 fatty alcohol compounds, C10-C20 saturated fatty acid compounds, epoxidized soybean oil, ethanol, silicone compounds and organic silicone oil;
the antifreeze agent can be one or more selected from sorbitol, ethylene glycol, polyethylene glycol, propylene glycol, glycerol, urea and sodium chloride;
the thickener can be one or more selected from gelatin, xanthan gum, polyethylene glycol and polyvinyl alcohol;
the filler can be one or more selected from light calcium carbonate, diatomite, bentonite, attapulgite and white carbon black;
the solvent may be selected from deionized water or methyl oleate.
If not specified, the additives can be routinely selected according to the preparation formulation of the bacillus microbial inoculum to be prepared, and the bacillus microbial inoculum can be prepared into the required preparation formulation.
Unless otherwise indicated, reagents, consumables, and the like, according to the present invention are commercially available, and are generally carried out under conventional conditions, or under conditions recommended by the reagent company, unless otherwise specified.
The invention also provides application of the bacillus inoculant in promoting plant growth. The brown algae oligosaccharide and the bacillus are jointly applied in the bacillus microbial inoculum provided by the invention, the brown algae oligosaccharide can provide nutrition for the bacillus, so that the bacillus can keep bioactivity and high survival capability under the condition of full nutrition after the microbial inoculum is applied, the sports capability of the bacillus can be obviously promoted by the brown algae oligosaccharide, the sports capability of the bacillus on soil or plant surfaces is obviously improved, and the bacillus can be quickly and effectively colonized on plant rhizosphere to play a role, and the effect of obviously promoting plant growth is realized.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Fucoidin promotes the movement ability of bacillus
1. Preparation of brown alginate oligosaccharides:
(1) Taking 5g of sodium alginate, and adding 100mL of distilled water;
(2) Adding alginic acid lyase to make its concentration in solution be 0.5g/L, its enzymolysis temperature is 30 deg.C, and its enzymolysis time is 16 hr;
(3) Centrifuging the enzymolysis solution (6000 rpm,5 min) to remove undegraded polysaccharide, and obtaining supernatant as brown algae oligosaccharide solution;
(4) And (3) performing vacuum freeze drying on the brown alginate oligosaccharide solution for 48 hours, and grinding to obtain the brown alginate oligosaccharide with the molecular weight of 1000-4000D.
2. Preparing a semisolid culture medium containing different kinds of sugar, wherein the semisolid culture medium comprises the following components: 10g/L peptone, 5g/LNaCl,0.7% agar, pH 7.0.+ -. 0.2, and sugar types and concentrations are shown in Table 1, wherein the brown alginate oligosaccharides added are those in step 1. 25mL of the prepared semi-solid culture medium is poured into a culture dish, dried in an ultra-clean bench and placed at room temperature for 12 hours for standby.
TABLE 1 different sources of sugar added to semi-solid Medium and their addition concentrations
3. Culturing Bacillus amyloliquefaciens Cas02 in LB liquid medium under 28 deg.C and 150rpm shaking culture for 12 hr, regulating fresh bacterial suspension to OD with sterile water 600 =0.3, 1.0 μl of the above bacterial liquid was aspirated and inoculated into the semisolid culture medium in step 2. In a stationary state, incubation was carried out at 37℃for 12 hours, photographing was carried out during the culture and the presence or absence of swimming phenomenon of the strain was observed.
As can be seen from the results of FIG. 1, when the semisolid culture medium is not inoculated with the strain, and 5g/L of chitosan oligosaccharide, enteromorpha polysaccharide, carrageenan oligosaccharide, carrageenan, sodium alginate and fucoidin are added into the semisolid culture medium respectively, after the strain is cultured for 12 hours, no obvious strain swimming phenomenon exists on the surface of the semisolid culture medium; as is clear from the results of FIG. 2, when brown alginate oligosaccharides were added to the semi-solid medium at concentrations of 1.0g/L,2.5g/L and 5.0g/L, respectively, apparent strain swimming phenomenon occurred on the surface of the medium in which the strain was cultured. When the adding concentration of the brown alginate oligosaccharides is 10.0g/L, no strain grows after 12 hours of culture after the strain is inoculated, i.e. the high concentration of the brown alginate oligosaccharides inhibits the normal growth of the strain. It is shown that the brown algae oligosaccharide can obviously promote the movement ability of the strain within a certain addition amount range, and is positively correlated with the concentration of the brown algae oligosaccharide.
Example 2
The embodiment provides a bacillus aqua, which comprises bacillus amyloliquefaciens and brown algae oligosaccharides.
The preparation method of the bacillus aqua comprises the following steps:
1. culturing Bacillus amyloliquefaciens in LB liquid medium at 28deg.C under 150rpm shaking culture for 12 hr, centrifuging (6000 rpm,5 min) to obtain thallus, suspending the thallus in sterile water, and regulating OD 600 =0.3, ready for use;
2. and (3) adding the brown alginate oligosaccharides obtained in the example 1 into the bacterial liquid obtained in the step (1), and regulating the concentration of the brown alginate oligosaccharides in the bacterial liquid to be 5g/L, thus obtaining the bacillus aqua.
Example 3
The difference from example 2 was that the brown alginate oligosaccharides concentration in the bacterial liquid was 2.5g/L.
Example 4
The only difference from example 2 is that the bacillus is bacillus subtilis.
Example 5
The only difference from example 2 is that the bacillus is bacillus belicus.
Example 6
Brown alginate oligosaccharides promote effective colonization of bacillus in plant rhizosphere
1. Culturing Bacillus amyloliquefaciens Cas02 in LB liquid medium at 28deg.C under 150rpm shaking culture for 12 hr, centrifuging (6000 rpm,5 min) to obtain thallus, suspending thallus in sterile water, and regulating OD 600 ＝0.3Standby;
2. sterilizing tobacco seeds, placing the tobacco seeds in a sterilized nutrition matrix for seedling culture, and transferring the tobacco seedlings into a nutrient solution containing MS when the tobacco seedlings grow to 4-5 leaves;
3. the nutrient solution for transplanting the tobacco seedlings is divided into three groups:
control group CK: adding 2mL of sterile water to the nutrient solution;
experimental group Cas02: 2mL of Bacillus amyloliquefaciens (OD) was applied to the nutrient solution 600 ＝0.3)；
Experimental group hsg+cas02: 2mL of the aqueous Bacillus aqua of example 2 was applied to the nutrient solution.
4. Tobacco seedlings are cultivated in a greenhouse (illumination condition: 16h,28 ℃ C.; darkness condition: 8h,20 ℃ C.; relative humidity: 60%) and after 4 days, the tobacco seedlings are taken out, roots are rinsed with sterilized water, and roots in a 1cm maturation zone are placed in a sterilized EP tube for observing the colonization condition of the strain in the rhizosphere by laser confocal. Wherein the laser confocal microscope model Leica TCS SP8 (Mannheim, germany), excitation wavelength 488, accept wavelengths 505-550.
As shown in fig. 3, the experiment group hsg+cas02 has the greatest number of bacillus colonizing the tobacco rhizosphere, which indicates that the brown alginate oligosaccharides in the bacillus microbial inoculum provided by the invention can significantly promote the colonization of bacillus in the plant rhizosphere.
Example 7
Promotion effect of alginate oligosaccharide and bacillus amyloliquefaciens combined application on plant growth
1. Sterilizing tobacco seeds, placing the tobacco seeds in a sterilized nutrition matrix for culturing, transferring the tobacco seedlings into a flowerpot (diameter is about 10 cm) containing about 180g of natural soil when the tobacco seedlings grow to 4-5 leaves, culturing the tobacco seedlings in a greenhouse (illumination condition: 16h,28 ℃ C.; darkness condition: 8h,20 ℃ C.; relative humidity: 60%) and irrigating 60 mL/plant of sterilizing water every week;
2. when the tobacco seedlings grow to 6-7 leaves, different treatments are carried out on the potted plants for planting the tobacco seedlings, and each treatment is repeated for 5 times: 20mL of sterile water (H) was evenly poured around the plants, respectively 2 O), 20mL of brown alginate oligosaccharides (HSG), 20mL of Bacillus amyloliquefaciens bacterial liquid (C), 20mL of Bacillus subtilis of example 2Aqueous (hsg+c); wherein, brown alginate oligosaccharides are prepared in example 1, and the concentration is 5g/L; the Bacillus amyloliquefaciens bacterial liquid is fresh bacterial liquid cultured by using LB culture medium, the bacterial liquid is centrifuged (6000 rpm,5 min) to obtain bacterial cells, the bacterial cells are suspended in sterile water, and OD is regulated 600 =0.3, culture method was the same as in example 2;
irrigation once every 7 days, twice in succession; and detecting the growth promoting effect of bacillus on tobacco seedlings 7 days after the second watering is finished. The experimental results are shown in fig. 4 and table 2.
TABLE 2 treatment of agronomic traits by different experiments
Note that: the data are averages of 5 replicates, different letters representing the same trait and differing significantly at the p < 0.05 level for different treatments.
As shown in Table 2, when the brown alginate oligosaccharides and bacillus are combined in the bacillus microbial inoculum provided by the invention, compared with the case that only brown alginate oligosaccharides and only bacillus amyloliquefaciens are used, the plant height, the maximum leaf length, the maximum leaf width, the effective leaf number and the maximum leaf area of tobacco are obviously increased, and the growth of tobacco seedlings is promoted; the combined administration of brown alginate oligosaccharides with bacillus increased the tobacco plant height by 44.44%, the maximum leaf length by 66.23%, the maximum leaf width by 56.22%, the effective leaf number by 14.42% and the maximum leaf area by 47.12% as compared to the control group irrigated with sterile water. It can be seen that the co-application of brown alginate oligosaccharides and bacillus can significantly promote the growth of plants.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (3)
1. The application of the brown algae oligosaccharide in promoting the movement of bacillus is characterized in that the preparation method of the brown algae oligosaccharide comprises the following steps: dissolving sodium alginate in water, adding enzyme for cleavage, centrifuging, and lyophilizing supernatant to obtain brown alginate oligosaccharide; the enzyme is alginic acid lyase, the enzymolysis temperature is 25-32 ℃, and the enzymolysis time is 12-18h; the molecular weight of the brown alginate oligosaccharides is 1000-4000D.
2. The application of the brown alginate oligosaccharides in promoting the colonization of bacillus plant rhizosphere is characterized in that the preparation method of the brown alginate oligosaccharides comprises the following steps: dissolving sodium alginate in water, adding enzyme for cleavage, centrifuging, and lyophilizing supernatant to obtain brown alginate oligosaccharide; the enzyme is alginic acid lyase, the enzymolysis temperature is 25-32 ℃, and the enzymolysis time is 12-18h; the molecular weight of the brown alginate oligosaccharides is 1000-4000D.
3. The use according to claim 1 or 2, wherein the bacillus is one or more of bacillus subtilis, bacillus belicus and bacillus amyloliquefaciens.
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