CN114806931A - Bacillus belgii YQ-1-8 and application thereof - Google Patents
Bacillus belgii YQ-1-8 and application thereof Download PDFInfo
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Abstract
The invention relates to the technical field of microorganisms, and particularly discloses a Bacillus beiLeisi YQ-1-8 and application thereof. The strain preservation number of the Bacillus velezensis YQ-1-8 is CGMCC No.24438, and the Bacillus velezensis YQ-1-8 can be used as a plant growth promoting and saline-alkali soil improving product. The Bacillus belgii YQ-1-8 provided by the invention has obvious saline-alkali soil improvement and plant growth promotion effects, can promote the rapid growth of plants in a facility degraded saline-alkali soil environment, and has important application value in the field of facility planting.
Description
Technical Field
The invention relates to the technical field of microorganisms, and particularly relates to Bacillus beiLeisi YQ-1-8 and application thereof.
Background
Facility planting is a modern agricultural production mode for ensuring annual supply of vegetables and improving agricultural planting benefits. However, the soil inside the facility is affected by factors such as closed shed room structure, excessive fertilization and blind irrigation, so that the problems of soil hardening, secondary salinization and the like often occur, and the yield and the quality of crops are reduced. Therefore, improvement and restoration of facility degraded soil are urgently needed to effectively guarantee the sustainable development of the facility vegetable industry.
Through years of research and practical application, rescue measures such as alternate stubble rotation, salt removal with water, soil replacement and salt reduction, organic fertilizer application increase and the like are established for improving degraded soil of facility vegetables, but the measures have great production limitation in intensive planting areas of the facility vegetables, the soil is difficult to be continuously improved, and the problem that plant growth is influenced due to degradation of the soil cannot be fundamentally solved. Therefore, finding new strategies to reduce the harm of soil salinization to crop growth is of great significance.
The microorganism has important effects on improving the physical and chemical properties of soil and supplying plant nutrients. The microbial agent with certain functions is combined with soil improvement measures, so that the degraded soil can be effectively improved, and the restoration of the degraded soil of the facility vegetables is more effective. The main manifestations of facility degraded soil are that the salinization degree of soil is increased and the plant growth is limited due to unbalanced nutrient supply, however, most microorganisms are difficult to grow in the degraded soil environment with high salinization degree, the action effect is not ideal, and the condition of plant growth limitation cannot be effectively improved.
Disclosure of Invention
Aiming at the problems of the existing facility degraded soil improvement measures, such as the problems, the invention provides the Bacillus beiLeisi YQ-1-8 and the application thereof, the Bacillus beiLeisi YQ-1-8 has good salt tolerance, can be normally propagated in the facility degraded soil with higher salinization degree, realizes good plant growth promotion effect in the salinization soil, and can ensure the normal growth of plants in the facility degraded soil.
In order to achieve the purpose of the invention, the embodiment of the invention adopts the following technical scheme:
the Bacillus subtilis YQ-1-8 has the strain preservation number of CGMCC No. 24438. The Bacillus beilaisi YQ-1-8 was deposited in China general microbiological culture Collection center (CGMCC) on 28 days 2.2022, and the deposition address is No.3 Siro No.1 Bichen of the sunward region in Beijing.
Compared with the prior art, the Bacillus belgii YQ-1-8 provided by the invention has the viability in a wide salt concentration environment, and meanwhile, the Bacillus belgii YQ-1-8 can effectively degrade organic phosphorus in soil, can generate a large amount of indole-3-acetic acid and extracellular polymer EPS, realizes the growth promotion effect on plants, and ensures that the plants can grow rapidly in salinized or hardened degraded soil. Therefore, the Bacillus belgii YQ-1-8 realizes the effect of promoting plant growth in facility degraded soil and has important application value in the field of facility planting.
The invention provides application of Bacillus beilesensis YQ-1-8 in improving products of saline-alkali soil.
The invention provides application of Bacillus belgii YQ-1-8 in plant production promotion products.
The invention provides application of Bacillus belgii YQ-1-8 in degradation of organic phosphorus products.
The invention provides application of Bacillus belgii YQ-1-8 in production of indole-3-acetic acid IAA.
The invention also provides application of the Bacillus belgii YQ-1-8 in production of extracellular polymeric substance EPS.
The invention also provides a microbial agent which comprises the Bacillus belgii YQ-1-8.
Preferably, the microbial agent further comprises a microbial carrier.
Preferably, the microorganism carrier comprises at least one of diatomite, attapulgite and sodium alginate.
The invention also provides a preparation method of the microbial agent, which comprises the following steps: activating the Bacillus belgii YQ-1-8, inoculating the activated Bacillus belgii YQ-1-8 into an LB liquid culture medium, and shaking and culturing at 25-28 ℃ for 20-30h to obtain a culture solution; then inoculating the culture solution into a fermentation culture medium, and shaking and culturing at 25-28 ℃ for 70-75h to obtain a fermentation solution; mixing the fermentation liquor with a microbial carrier, and drying to obtain the viable bacteria with the viable bacteria concentration of 2 × 10 8 cfu/g-2×10 9 cfu/g of a microbial agent; the formula of each liter of the fermentation medium is as follows: peptone 7-9g, glucose 8-12g, beef extract 4-5g, yeast powder 0.8-1.2g, and waterMake up to 1000mL, adjust the pH value to 6.8-7.2.
Drawings
FIG. 1 is a diagram showing the growth of cucumber seeds treated with YQ-1-8 bacteria solution and cucumber seeds of a blank control group in example 2 of the present invention;
FIG. 2 is a colony morphology feature map of strain YQ-1-8 in example 6 of the present invention;
FIG. 3 is a phylogenetic tree construction diagram of Bacillus beijerinckii YQ-1-8 in example 6 of the present invention;
FIG. 4 is a graph showing the results of organophosphorus decomposition test conducted by strain YQ-1-8 in example 7 of the present invention;
FIG. 5 is a graph showing the results of the IAA-producing ability test of strain YQ-1-8 in example 7 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The experimental procedures used in the following examples are all conventional in the art unless otherwise specified.
The raw materials, reagents and the like used in the following examples were obtained commercially unless otherwise specified.
In the following examples:
the LB solid culture medium has the formula: 10g of tryptone, 5g of yeast powder, 10g of NaCl, 15g of agar and distilled water are added to 1L, the pH is adjusted to 7.0, and the mixture is sterilized at 115 ℃ for 30 min.
The LB liquid culture medium has the formula: 10g of tryptone, 5g of yeast powder, 10g of NaCl and distilled water are added to 1L, the pH value is adjusted to 7.0, and the mixture is sterilized for 30min at 115 ℃.
The formula of the fermentation medium is as follows: 8g of peptone, 10g of glucose, 4.5g of beef extract, 1g of yeast powder and distilled water, wherein the distilled water is added to 1L, the pH value is adjusted to 7.0, and the mixture is sterilized at 115 ℃ for 30 min.
The potassium-dissolving culture medium comprises the following components in percentage by weight: sucrose 5g, Na 2 HPO 4 2g，MgSO 4 0.5g，FeCl 3 0.005g，CaCO 3 0.1g, potassium feldspar powder (300 meshes) 1g, agar 15g, distilled water to 1L, and sterilizing at 115 deg.C for 30 min.
The formula of the inorganic phosphorus culture medium is as follows: glucose 10g, (NH) 4 ) 2 SO 4 0.5g, yeast powder 0.5g, NaCl 0.3g, MgSO 4 0.3g，KCl 0.3g，FeSO 4 0.03g，MnSO 4 0.03g，Ca 3 (PO 4 ) 2 5g, agar 15g, distilled water to 1L, and sterilizing at 115 deg.C for 30 min.
The formula of the organophosphorus culture medium is as follows: glucose 10g, (NH) 4 ) 2 SO 4 0.5g, yeast powder 0.5g, NaCl 0.3g, MgSO 4 0.3g，KCl 0.3g，FeSO 4 0.03g，MnSO 4 0.03g, lecithin 0.2g, CaCO 3 1g of agar, 15g of agar and distilled water, and sterilizing at 115 ℃ for 30 min.
The formula of the IAA culture medium is as follows: k 2 HPO 4 1.15g, peptone 20g, glycerol 15ml, L-tryptophan 0.1g, MgSO 4 1.5g, adding distilled water to 1L, and sterilizing at 115 deg.C for 30 min.
The formula of the siderophore culture medium (CAS culture medium) is as follows: 60.5mg of chrome azure, 72.9mg of hexadecyl trimethyl ammonium bromide (HDTMA), FeCl 3 2.645g, 295.25mg of sodium dihydrogen phosphate dihydrate, 1213.5mg of sodium dihydrogen phosphate dodecahydrate, NH 4 Cl 125mg，KH 2 PO 4 37.5mg, NaCl 62.5mg, agar 9g, distilled water make up to 1L, and sterilize at 116 deg.C for 30 min.
The formula of the nitrogen fixation culture medium is as follows: KH (Perkin Elmer) 2 PO 4 0.2g，MgSO 4 0.2g，NaCl 0.2g，CaCO 3 5g, mannitol 10g, CaSO 4 0.1g, agar 15g, distilled water to 1L, and sterilizing at 121 deg.C for 15 min.
The formula of the EPS production culture medium is as follows: sucrose 20g, K 2 HPO 4 0.2g，KH 2 PO 4 0.5g，NaCl 100g，MgSO 4 0.5g, yeast powder 3g, distilled water to 1L, sterilizing at 121 deg.C for 15 min.
Example 1
Screening process for Bacillus belgii YQ-1-8
Using five-point sampling32 Handan perpetual years, corridor perpetual clear and cangzhou Suanning facility soil samples of more than ten years are collected in China. Screening the salt-tolerant microorganisms by adopting an LB culture medium with the salt content of 10 wt%. 5g of soil to be tested is added into a conical flask filled with 45mL of sterile water, and the mixture is shaken at 200r/min for 30min to obtain soil mixed liquor. Diluting the soil mixed solution according to gradient, and selecting the diluted concentration gradient as 10 -4 、10 -5 、10 -6 The soil mixed solution of (2) is respectively absorbed by 100 mul of the soil mixed solution and coated in LB solid culture medium containing 10 wt% of NaCl for separating the salt-tolerant microorganisms, and each dilution degree is 3 flat plates. The plate is placed upside down and cultured for 96 hours in a constant temperature incubator at 28 ℃, strains with different colony forms and larger colony diameters are selected on the plate, the strains are subjected to streaking purification for 3 times, 40 strains are obtained through co-purification, the serial numbers are shown in table 1, and the purified strains are stored in a refrigerator at-80 ℃ by using 25% of glycerol for later use.
Example 2
Primary screen for growth promoting effect of salt-tolerant strain
The 40 salt-tolerant strains stored at-80 ℃ in example 1 were activated on an LB plate, and then the activated strains were inoculated into an LB liquid medium and cultured at 28 ℃ for 24 hours. Centrifuging the bacterial liquid at 10000rpm for 10min, removing supernatant, adding 10ml of sterile water, shaking, re-suspending the thallus, and re-suspending the thallus with the sterile water until the OD600 is 0.1 to obtain the bacterial liquid. The cucumber seeds are placed in the bacterial liquid with the OD600 of 0.1 to be soaked for 4 hours, the cucumber seeds are placed on sterilized filter paper in order, 4mL of sterile saline containing 1 wt% of NaCl is added (the sterile saline is fully soaked), the mixture is placed in an incubator at 28 ℃ to be cultured for 72 hours, the simplified activity index growth rate of the cucumber seeds is measured, and the growth promotion result of the strains on the cucumbers is counted (the cucumber seeds which are not treated by the bacterial liquid in the same batch are used as blank control).
Simplified vigor index (average root length + average stem length) x germination rate of seeds
Simplified vitality index increase rate ═ simplified vitality index of treatment group-simplified vitality index of control group)/simplified vitality index of control group × 100%
As shown in Table 1, 10 strains such as 1A-3, 2A-2 and YN4-2 showed no growth promoting effect, and 21 strains such as 2A-1, 2A-7 and YQ-3-1 showed a simplified activity index increase rate of 0 to 10%. The simplified activity index growth rate of 9 strains of 3A-3, YQ-1-3, SN-1-5 and the like is more than 10 percent, wherein the simplified activity index growth rate of the YQ-1-8 strain is the highest and reaches 15.18 percent.
TABLE 1 reduction of the Activity index growth Rate of the strains%
| Strain numbering | Rate of increase/%) | Strain numbering | Rate of increase/%) | Strain numbering | Rate of increase/%) |
| 1A-3 | - | YN-2-3 | - | SN-1-2 | 10.75 |
| 2A-1 | 9.45 | YN-2-6 | 2.79 | SN-1-5 | 10.62 |
| 2A-2 | - | YN-2-2 | 8.36 | YQ-8-3 | 2.79 |
| 2A-7 | 7.82 | YN-3-6 | 1.33 | YQ-8-4 | 7.96 |
| 3A-3 | 11.40 | YN-3-4 | 2.92 | YQ-1-1 | 4.88 |
| YQ-1-3 | 11.89 | SN-4-2 | 5.04 | YQ-1-4 | 2.66 |
| YQ-3-1 | 4.07 | YN-4-3 | - | YQ-1-5 | 13.58 |
| SN-1-6 | 2.44 | SN-4-4 | - | YQ-8-1 | 10.18 |
| SN-13-3 | 5.70 | SN-13-2 | - | YQ-1-8 | 15.18 |
| YN-4-2 | - | YN-4-5 | 3.45 | YQ3-2 | 10.19 |
| 1A-2 | - | SN-4-3 | - | SN-4-6 | 8.81 |
| YQ-3-3 | 2.78 | YQ-1-7 | - | SN-13-4 | 12.92 |
| YQ-3-4 | 6.07 | YQ-8-2 | 9.51 | ||
| YN-3-2 | 0.54 | YN-3-5 | 2.36 |
Wherein, the growth conditions of the cucumber seeds treated by the YQ-1-8 bacteria liquid and the cucumber seeds of the blank control group are shown in figure 1.
Example 3
The salt tolerance of the 9 strains with simplified activity exponential growth rate of more than 10 percent obtained by screening in example 2 is determined
Activating 9 strains with growth promoting capacity of more than 10% on an LB culture medium, then inoculating the activated strains on an LB solid culture medium with NaCl mass concentration of 5%, 7%, 10%, 15% and 20%, culturing at constant temperature of 28 ℃ for 7d, repeating each strain for 3 times, and observing the growth condition of the strains under different salt concentration gradients. Very vigorous growth is marked as "+ + + + +", vigorous growth is marked as "+ +", more vigorous growth is marked as "+", general growth is marked as "+", and inhibited growth is marked as "-". As shown in Table 2, it can be seen that the salt tolerance of YQ-1-3, YQ-8-1, YQ-3-2, SN-13-4, YQ-1-8 and 3A-3 among the 9 growth-promoting strains is strong, wherein the YQ-1-8 strain can still grow normally in LB medium with salt content of 20%.
TABLE 2 growth of growth-promoting bacteria at different salt concentrations
| Strain numbering | 5％ | 7％ | 10％ | 15％ | 20％ |
| YQ-1-3 | +++ | ++ | ++ | + | - |
| YQ-1-5 | ++ | ++ | + | - | - |
| YQ-1-8 | ++ | ++ | ++ | ++ | ++ |
| YQ-8-1 | +++ | ++ | ++ | + | - |
| YQ-3-2 | ++ | ++ | ++ | + | - |
| SN-1-2 | ++++ | ++++ | ++ | - | - |
| SN-1-5 | ++++ | ++++ | ++ | - | - |
| SN-13-4 | ++++ | ++++ | ++ | + | - |
| 3A-3 | ++ | ++ | ++ | + | - |
Example 4
Selecting strains 3A-3 and YQ-1-8 with good growth promoting effect and good salt tolerance, and determining the soil colonization ability
Activating the YQ-1-8 strain, inoculating the activated strain in an LB liquid culture medium, and shaking at 26 ℃ for 24 hours to obtain a culture solution; then inoculating the culture solution into a fermentation culture medium, and shaking and culturing for 72 hours at 26 ℃ to obtain a fermentation solution; mixing the fermentation liquid with diatomaceous earth, and spray drying to obtain viable bacteria with concentration of 1 × 10 9 cfu/g microbial agent.
A microbial agent comprising the 3A-3 strain was obtained in the same manner.
The determination of the colonization of the 3A-3 strain and the YQ-1-8 strain in the soil was determined by a potting test. Cucumber was selected as a cultivated crop and the experiment was carried out in a flowerpot with a diameter of about 7cm and a height of 7 cm. As shown in Table 3, it was found that the colonization ability of the strain YQ-1-8 was strong and the number of viable bacteria in the soil was small in the amounts of the strains 1, 3, 5, 7, 14, 28 and 60d, as shown in Table 3, when the microbial agent and the sterilized soil were mixed (the viable bacteria concentration in the soil after mixing is shown in Table 3) and then the viable bacteria counts of the strain 3A-3 and the strain YQ-1-8 in the soil were measured at 1d, 3d, 5d, 7d, 14d, 28d and 60d after inoculation, respectively. The strain YQ-1-8 has good soil colonization ability, and the number of the strain YQ-1-8 in the soil is 2.19 multiplied by 10 after inoculation for 60 days 7 cfu/g, can be used as a microbial preparation or a fertilizer for application on crops.
TABLE 3 colonization of the strains in the soil
| Inoculation time (d) | Strain YQ-1-8 (x 10) 7 cfu/g) | Strain 3A-3(× 10) 7 cfu/g) |
| 1 | 2.61 | 16.53 |
| 3 | 2.06 | 10.12 |
| 5 | 0.94 | 6.20 |
| 7 | 1.46 | 2.44 |
| 14 | 1.53 | 0.89 |
| 28 | 1.68 | 0.90 |
| 60 | 2.19 | 0.85 |
Example 5
Growth promoting effect of salt-tolerant growth promoting strain YQ-1-8 on cucumber
Streaking and activating a strain YQ-1-8 on an LB solid culture medium, culturing for 48h at 28 ℃, selecting lawn, inoculating the lawn into 100mL of sterilized LB liquid culture medium, culturing for 12h under the conditions of a shaking table at 28 ℃ and 200rpm/min, centrifuging bacterium liquid at 10000rpm for 10min, removing supernatant, adding 10mL of sterile water, shaking, re-suspending the thallus with the sterile water and adjusting OD (optical density) of the re-suspended thallus 600 The bacterial suspension was obtained at 0.1.
The preparation method of the bacterial liquid of the 3A-3 bacterial strain is the same as the above.
Performing pot culture test, adjusting the soil to 0.5% of salt content, adding the soil into a flowerpot with diameter of 7cm and height of 7cm, and testing the soil and concentration of OD 600 Mixing 0.1 bacterial liquid (10kg soil: 1L bacterial liquid), using a treatment of adding an equal amount of sterile water as a control group, planting cleaned cucumber seeds with the same size in a flowerpot, 3 seeds per pot, keeping seedlings with the same growth vigor after the cucumber sprouts, keeping 1 cucumber seedling per pot, and measuring the stem thickness, the plant height, the overground fresh quality, the dry quality and the chlorophyll content of the cucumber after culturing for 4 weeks. As shown in Table 4, YQ-1-8 can promote the growth of cucumber in saline soil more obviously than 3A-3 strain.
TABLE 4 Effect of the strains on cucumber seedling growth
Note: data are represented by mean ± Standard Deviation (SD), with different lower case letters indicating p <0.05 level difference.
Example 6
Identification of Strain YQ-1-8
Morphological identification:
the strain YQ-1-8 is taken out from a low-temperature refrigerator and inoculated on an LB solid medium, the strain is cultured for 48 hours in an incubator at the temperature of 28 ℃, and the growth condition and morphological characteristics of the colony are observed and shown in figure 2: the strain grows fast, and is oval, white, opaque, convex and wrinkled at the edge.
Molecular biological identification:
inoculating the strain numbered YQ-1-8 to LB culture medium, shaking-culturing at 28 ℃ for 24h, extracting the genomic DNA of the strain by adopting a genomic DNA extraction kit of Tiangen Biochemical technology (Beijing) Co., Ltd, and then carrying out PCR amplification by using the extracted total DNA as a template and 27f (5'-AGAGTTTGATCCTGGCTCAG-3', SEQ ID NO.1) and 1492r (5'-TACGGCTACCTTGTTACGACTT-3', SEQ ID NO.2) primer pairs. The PCR reaction system is 25 mu L, and the reaction system is as follows: genomic DNA 1. mu.L, 10 XPCR buffer 2.5. mu.L, 27f primer 0.5uL, 1492r primer 0.5 uL, d NTPs 2 uL, Taq enzyme (5U/. mu.L) 0.5 uL, ddH 2 O make up 25. mu.L. The reaction conditions are as follows: pre-denaturation at 94 deg.C for 5 min; denaturation at 94 deg.C for 1min, annealing at 50 deg.C for 1min, extension at 72 deg.C for 2min, and circulation for 35 times; extension at 72 ℃ for 10 min. The PCR product was detected by electrophoresis using 1% agarose gel, and the 16S rDNA sequence (shown in SEQ ID NO. 3) was obtained by sequencing the PCR product with a PCR band size of about 1500bp by Shanghai Biotechnology Ltd.
The obtained 16S rDNA gene sequence is subjected to homology alignment analysis by logging in an NCBI website (www.ncbi.nlm.nih.gov), sequences with similar homology are selected for phylogenetic analysis, multiple alignment of the sequences is firstly carried out, then a phylogenetic tree is constructed by using MEGA 7 software and adopting an adjacency method (neighbor-Joining), and finally the phylogenetic position of the strain is determined as shown in figure 3. Sequence analysis shows that the strain belongs to a strain of Bacillus velezensis (Bacillus velezensis). The Bacillus belgii YQ-1-8 is preserved in China general microbiological culture Collection center (CGMCC) at 28 days 2.2022, with the culture preservation number of CGMCC No.24438 and the preservation address of No.3 Hospital No.1 Samura No.3 of the morning-Yang district of Beijing.
Example 7
Analysis of growth promoting mechanism of strain YQ-1-8:
and (3) streaking the preliminarily screened strain YQ-1-8 on an LB solid culture medium, and carrying out inverted culture in an incubator at 28 ℃ for 2d for later use. And inoculating the activated strain into 50mL of LB liquid culture medium, and performing shake culture at 28 ℃ for 24h to obtain a bacterial liquid required by the test. And respectively inoculating 5 mu L of the bacterial liquid to prepared potassium dissolving culture medium, inorganic phosphorus culture medium, organic phosphorus culture medium, siderophore culture medium and nitrogen fixation culture medium plates, culturing for 7d at the temperature of 28 ℃, and observing and recording test results, wherein the test results are shown in Table 5. The strain grows on an organophosphorus culture medium and generates turbid spots around the strain, and the turbid spots have the capacity of degrading organophosphorus in soil as shown in a figure 4.
Inoculating the bacterial solution into IAA culture medium at 5%, shake culturing at 28 deg.C for 48 hr, centrifuging at 10000 Xg for 10min, collecting supernatant 1ml and Salkowski reagent 2ml (concentrated sulfuric acid 15ml, double distilled water)25ml，0.5mol/l FeCl 3 0.75ml) and then dark for 30min, and the color of the IAA medium was observed to turn pink, as shown in FIG. 5, indicating that the strain has IAA producing ability. In addition, inoculating the strain to EPS culture medium according to the inoculation amount of 5%, performing shake culture at 28 ℃ for 48h, centrifuging 10ml of culture solution at 10000r/min for 10min, removing supernatant, and drying and weighing thalli; and simultaneously, adding 30mL of 95% ethanol into the supernatant for alcohol precipitation, standing in a refrigerator at 4 ℃ overnight, centrifuging the mixed solution at 10000r/min for 10min, removing the supernatant, drying and weighing the precipitate, wherein the ratio of the EPS dry weight to the thallus dry weight is the EPS yield of the strain, and the determination result shows that the strain can produce extracellular polymers with the production quantity of 0.467 g/g.
TABLE 5 analysis of growth promoting mechanism of strain YQ-1-8
| Kind of growth promoting mechanism | Growth promoting effect |
| Decomposing potassium | - |
| Inorganic phosphorus removal | - |
| Organic phosphorus removal | + |
| Produce IAA | + |
| Iron production carrier | - |
| Fixation of nitrogen | - |
| Produce EPS | +(0.467g/g) |
Note: "+": has the function; "-": without this function.
Example 8
Effect of Bacillus beleisi YQ-1-8 microbial inoculum on improvement of salinized soil
Compounding Bacillus beiLeisi YQ-1-8 microbial inoculum with attapulgite to prepare microbial inoculum with effective bacteria number of 2 hundred million (activating Bacillus beiLeisi YQ-1-8, inoculating in LB liquid culture medium, shaking at 26 deg.C for 24 hr to obtain culture solution, inoculating the culture solution into fermentation culture medium, shaking at 26 deg.C for 72 hr to obtain fermentation broth, mixing the fermentation broth with attapulgite, and drying to obtain viable bacteria concentration of 2 × 10 8 cfu/g microbial agent) for soil improvement test. The test is carried out by adopting conventional management and applying three treatments of YQ-1-8 microbial inoculum and attapulgite (a control group), cucumber is taken as a test crop, the test is carried out in a greenhouse with the planting time of more than 10 years, the YQ-1-8 microbial inoculum is applied before land preparation, the dosage per mu is 6 kg/mu, and each treatment is repeated for 3 times and is arranged randomly. And (3) detecting soil salinity improvement indexes of the saline-alkali soil 30 days after the cucumber is transplanted, measuring the total salt content of the soil by adopting a leaching agent drying method, measuring the conductivity of the soil by adopting a conductivity meter, and measuring the yield of the cucumber in a harvest period. As shown in Table 6, the soil salinity can be reduced by 11.39%, the conductivity can be reduced by 16.38%, and the yield can be improved by 11.91% by applying the Bacillus beijerinckii YQ-1-8 microbial inoculum. The YQ-1-8 microorganism has better effects of reducing the soil salt content and improving the crop yield.
TABLE 6 influence of YQ-1-8 on soil salinity and yield
| Treatment of | Total salt content (g/kg) | Conductivity (μ S/cm) | Yield (kg/mu) |
| Routine management | 8.95 | 460.2 | 7424 |
| YQ-1-8 | 7.93 | 384.8 | 8308 |
| Attapulgite | 8.12 | 440.3 | 7942 |
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents or improvements made within the spirit and principle of the present invention should be included in the scope of the present invention.
SEQUENCE LISTING
<110> university of Hebei science and technology
<120> Bacillus belgii YQ-1-8 and application thereof
<130> 2022
<160> 3
<170> PatentIn version 3.5
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agagtttgat cctggctcag 20
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<212> DNA
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tacggctacc ttgttacgac tt 22
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<212> DNA
<213> 16S rDNA
<400> 3
gcgggcgcgt gctatacatg caagtcgagc ggacagatgg gagcttgctc cctgatgtta 60
gcggcggacg ggtgagtaac acgtgggtaa cctgcctgta agactgggat aactccggga 120
aaccggggct aataccggat ggttgtttga accgcatggt tcagacataa aaggtggctt 180
cggctaccac ttacagatgg acccgcggcg cattagctag ttggtgaggt aacggctcac 240
caaggcaacg atgcgtagcc gacctgagag ggtgatcggc cacactggga ctgagacacg 300
gcccagactc ctacgggagg cagcagtagg gaatcttccg caatggacga aagtctgacg 360
gagcaacgcc gcgtgagtga tgaaggtttt cggatcgtaa agctctgttg ttagggaaga 420
acaagtgccg ttcaaatagg gcggcacctt gacggtacct aaccagaaag ccacggctaa 480
ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg ttgtccggaa ttattgggcg 540
taaagggctc gcaggcggtt tcttaagtct gatgtgaaag cccccggctc aaccggggag 600
ggtcattgga aactggggaa cttgagtgca gaagaggaga gtggaattcc acgtgtagcg 660
gtgaaatgcg tagagatgtg gaggaacacc agtggcgaag gcgactctct ggtctgtaac 720
tgacgctgag gagcgaaagc gtggggagcg aacaggatta gataccctgg tagtccacgc 780
cgtaaacgat gagtgctaag tgttaggggg tttccgcccc ttagtgctgc agctaacgca 840
ttaagcactc cgcctgggga gtacggtcgc aagactgaaa ctcaaaggaa ttgacggggg 900
cccgcacaag cggtggagca tgtggtttaa ttcgaagcaa cgcgaagaac cttaccaggt 960
cttgacatcc tctgacaatc ctagagatag gacgtcccct tcgggggcag agtgacaggt 1020
ggtgcatggt tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg caacgagcgc 1080
aacccttgat cttagttgcc agcattcagt tgggcactct aaggtgactg ccggtgacaa 1140
accggaggaa ggtggggatg acgtcaaatc atcatgcccc ttatgacctg ggctacacac 1200
gtgctacaat ggacagaaca aagggcagcg aaaccgcgag gttaagccaa tcccacaaat 1260
ctgttctcag ttcggatcgc agtctgcaac tcgactgcgt gaagctggaa tcgctagtaa 1320
tcgcggatca gcatgccgcg gtgaatacgt tcccgggcct tgtacacacc gcccgtcaca 1380
ccacgagagt ttgtaacacc cgaagtcggt gaggtaacct tttaggagcc agccgccgaa 1440
ggtacaga 1448
Claims (10)
1. The Bacillus belgii YQ-1-8 is characterized in that: the strain preservation number of the Bacillus velezensis YQ-1-8 is CGMCC No. 24438.
2. The use of Bacillus belgii YQ-1-8 according to claim 1 as a saline-alkali soil improvement product.
3. Use of Bacillus belgii YQ-1-8 according to claim 1 as a plant growth promoting product.
4. Use of Bacillus belgii YQ-1-8 according to claim 1 as a product for degrading organophosphorus.
5. Use of Bacillus belgii YQ-1-8 according to claim 1 for the production of indole-3-acetic acid IAA.
6. Use of Bacillus belgii YQ-1-8 according to claim 1 for the production of extracellular polymer EPS.
7. A microbial inoculant characterized by: comprising Bacillus belgii YQ-1-8 as claimed in claim 1.
8. The microbial inoculant according to claim 7, wherein: also comprises a microorganism carrier.
9. The microbial inoculant according to claim 8, wherein: the microbial carrier comprises at least one of diatomite, attapulgite and sodium alginate.
10. The process for the preparation of a microbial inoculant according to any one of claims 7 to 9, wherein: the method comprises the following steps: activating the Bacillus belgii YQ-1-8, inoculating the activated Bacillus belgii YQ-1-8 into an LB liquid culture medium, and shaking and culturing at 25-28 ℃ for 20-30h to obtain a culture solution; then inoculating the culture solution into a fermentation culture medium, and shaking and culturing at 25-28 ℃ for 70-75h to obtain a fermentation solution; mixing the fermentation liquor with a microbial carrier, and drying to obtain the viable bacteria with the viable bacteria concentration of 2 × 10 8 cfu/g-2×10 9 cfu/g of a microbial agent; the formula of each liter of the fermentation medium is as follows: 7-9g of peptone, 8-12g of glucose, 4-5g of beef extract and 0.8-1.2g of yeast powder, supplementing water to 1000mL, and adjusting the pH value to 6.8-7.2.
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| CN115820461A (en) * | 2022-08-11 | 2023-03-21 | 上海市农业科学院 | High-yield indoleacetic acid bacterial strain JB0319 and application thereof |
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