CN110684696A - Bacillus megaterium QZY-3 and application thereof - Google Patents
Bacillus megaterium QZY-3 and application thereof Download PDFInfo
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- 241000194107 Bacillus megaterium Species 0.000 title claims abstract description 56
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims abstract description 31
- 229910052698 phosphorus Inorganic materials 0.000 claims abstract description 31
- 239000011574 phosphorus Substances 0.000 claims abstract description 31
- 230000012010 growth Effects 0.000 claims abstract description 24
- 238000000855 fermentation Methods 0.000 claims abstract description 22
- 230000004151 fermentation Effects 0.000 claims abstract description 22
- 230000001737 promoting effect Effects 0.000 claims abstract description 12
- 238000002360 preparation method Methods 0.000 claims abstract description 7
- 238000004321 preservation Methods 0.000 claims abstract description 7
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims abstract description 6
- 239000001506 calcium phosphate Substances 0.000 claims abstract description 6
- 239000000787 lecithin Substances 0.000 claims abstract description 6
- 235000010445 lecithin Nutrition 0.000 claims abstract description 6
- 229940067606 lecithin Drugs 0.000 claims abstract description 6
- 239000002367 phosphate rock Substances 0.000 claims abstract description 6
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 claims abstract description 6
- 229940078499 tricalcium phosphate Drugs 0.000 claims abstract description 6
- 229910000391 tricalcium phosphate Inorganic materials 0.000 claims abstract description 6
- 235000019731 tricalcium phosphate Nutrition 0.000 claims abstract description 6
- 239000003337 fertilizer Substances 0.000 claims abstract description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- 239000001963 growth medium Substances 0.000 claims description 16
- 239000011780 sodium chloride Substances 0.000 claims description 9
- 229920001817 Agar Polymers 0.000 claims description 8
- 239000001888 Peptone Substances 0.000 claims description 8
- 108010080698 Peptones Proteins 0.000 claims description 8
- 239000008272 agar Substances 0.000 claims description 8
- 235000015278 beef Nutrition 0.000 claims description 8
- 239000012153 distilled water Substances 0.000 claims description 8
- 235000019319 peptone Nutrition 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 7
- 239000002609 medium Substances 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 4
- 239000008103 glucose Substances 0.000 claims description 4
- 239000000654 additive Substances 0.000 claims description 3
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L magnesium chloride Substances [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims description 3
- 229910001629 magnesium chloride Inorganic materials 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- CSNNHWWHGAXBCP-UHFFFAOYSA-L magnesium sulphate Substances [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 3
- 239000003895 organic fertilizer Substances 0.000 claims description 3
- 230000003381 solubilizing effect Effects 0.000 claims description 3
- 238000003860 storage Methods 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 239000002689 soil Substances 0.000 abstract description 17
- 241000209140 Triticum Species 0.000 abstract description 8
- 235000021307 Triticum Nutrition 0.000 abstract description 8
- 230000000813 microbial effect Effects 0.000 abstract description 4
- 244000005700 microbiome Species 0.000 abstract description 3
- 239000002054 inoculum Substances 0.000 abstract description 2
- 239000008223 sterile water Substances 0.000 description 9
- 241000894006 Bacteria Species 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 108020004465 16S ribosomal RNA Proteins 0.000 description 6
- 240000002791 Brassica napus Species 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 235000011293 Brassica napus Nutrition 0.000 description 4
- 229910019142 PO4 Inorganic materials 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 235000021317 phosphate Nutrition 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 230000001502 supplementing effect Effects 0.000 description 4
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 3
- 239000010452 phosphate Substances 0.000 description 3
- 239000002686 phosphate fertilizer Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- 241001052560 Thallis Species 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 230000004899 motility Effects 0.000 description 2
- 230000008092 positive effect Effects 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 238000009736 wetting Methods 0.000 description 2
- 235000006008 Brassica napus var napus Nutrition 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010014251 Muramidase Proteins 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 102000015439 Phospholipases Human genes 0.000 description 1
- 108010064785 Phospholipases Proteins 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- WYWFMUBFNXLFJK-UHFFFAOYSA-N [Mo].[Sb] Chemical compound [Mo].[Sb] WYWFMUBFNXLFJK-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 230000009604 anaerobic growth Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 230000015784 hyperosmotic salinity response Effects 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 229960000274 lysozyme Drugs 0.000 description 1
- 235000010335 lysozyme Nutrition 0.000 description 1
- 239000004325 lysozyme Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- CEQFOVLGLXCDCX-WUKNDPDISA-N methyl red Chemical compound C1=CC(N(C)C)=CC=C1\N=N\C1=CC=CC=C1C(O)=O CEQFOVLGLXCDCX-WUKNDPDISA-N 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 231100000027 toxicology Toxicity 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
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Abstract
The invention belongs to the technical field of microorganisms, and particularly relates to a bacillus megaterium QZY-3 and application thereof; the bacillus megaterium QZY-3 is preserved in China center for type culture Collection with the preservation number of CCTCC NO: m2019717; the invention provides a preparation method of strain fermentation liquor; the invention also provides the application of the bacillus megaterium QZY-3 in dissolving insoluble phosphorus and promoting the growth of crops; the bacillus megaterium QZY-3 has obvious dissolving capacity on inorganic insoluble phosphorus (tricalcium phosphate and ground phosphate rock) and organic insoluble phosphorus (lecithin), and has extremely obvious growth promoting effect on wheat seedlings and rape seedlings. Has good application prospect in the aspects of researching and developing microbial growth promoting inoculants, microbial fertilizers, popularization and demonstration on soil and the like.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to bacillus megaterium QZY-3 and application thereof.
Background
Phosphorus is the most important for crop growthOne of the elements. Available phosphorus directly absorbed by crops in chemical phosphate fertilizer is easy to be mixed with Ca in soil2+、Fe3+、Al3+Plasma is combined and converted into insoluble phosphate which can not be utilized by plants, so that the utilization rate of the chemical phosphate fertilizer is low, the conversion of the insoluble phosphate in soil into effective phosphorus which can be absorbed by plants is a hotspot of current research, the phosphorus-dissolving bacteria can convert insoluble phosphate in soil into the effective phosphorus, and the phosphorus-dissolving bacteria have positive effects on the soil phosphorus validation process; through the action of phosphorus-dissolving bacteria, the phosphorus-dissolving bacteria have positive effects on the aspects of improving the utilization rate of chemical phosphate fertilizer, increasing the content of available phosphorus in soil, reducing phosphorus accumulation and the like. The application research of the phosphorus-dissolving bacteria firstly needs to have efficient and safe strain resources. The bacillus megaterium belongs to the first level in the appendix A-strain safety classification catalogue of the biological safety general technical criteria (NY/T1109-.
The yellow soil is most widely distributed in Guizhou, 25.3 percent of yellow soil in China is intensively distributed in Guizhou, the area of the yellow soil accounts for 41.9 percent and 46.4 percent of the area of soil in Guizhou, the yellow soil is the main agricultural soil type in Guizhou and plays an important role in Guizhou agricultural production. At present, the research on the regional phosphorus-dissolving bacteria is less, and the efficient and safe phosphorus-dissolving strain is lack of resources, so that the method has important significance for screening and separating the phosphorus-dissolving strain from the soil (yellow soil) of the farmland in the middle of Guizhou and providing high-quality and safe strain resources for the research on the regional phosphorus-dissolving microorganisms.
Disclosure of Invention
The invention aims to provide bacillus megaterium QZY-3 which is preserved in China center for type culture Collection with the preservation number of CCTCC NO: m2019717.
The Bacillus megaterium QZY-3(Bacillus megaterium QZY-3) is separated and screened from yellow soil farmland in Guizhou, is preserved in China Center for Type Culture Collection (CCTCC) in 2019, 9, 12 and has the following addresses: wuhan, Wuhan university; the preservation number is: CCTCC NO: m2019717. It has the following biological properties: culturing the strain on a beef extract peptone culture medium (3 g of beef extract, 10g of peptone, 5g of NaCl, 15g of agar and 1000ml of distilled water) consisting of the following components in parts by mass/volume at a constant temperature of 32 ℃ for 24 hours, wherein the pH value of the culture solution is 7.0-7.2, the strain forms a large number of spores, forms long rods (1.2-1.5 multiplied by 2.8-4.0 mu m) of the strain before the spores, and is arranged into single or chain (shown in figure 1) and is gram-positive; after 3 days of culture, the colony morphology is as follows: the bacterial colony is large and round, milky white, neat in edge, smooth and moist in surface and non-transparent (see figure 2); the puncture inoculation cultured thalli grows in a clouded and misty spread mode in a culture medium of a test tube, and the motility is shown.
The invention also provides a preparation method of the strain fermentation liquor, which comprises the following steps: taking a preservation number of CCTCC NO: inoculating the Bacillus megaterium QZY-3 of M2019717 into a sterilized culture medium for fermentation, and culturing at 32 ℃ and 160r/min for 24 hours to obtain a fermentation liquid; wherein the storage and fermentation culture medium of the bacillus megaterium QZY-3 comprises 3.0g of beef extract, 10.0g of peptone, 5.0g of NaCl and 1000ml of distilled water, 15g of agar is added into a solid culture medium, and the pH is adjusted to 7.0-7.2.
The invention also provides application of the bacillus megaterium QZY-3 in dissolving insoluble phosphorus.
Further, the application of the bacillus megaterium QZY-3 in dissolving the insoluble phosphorus is realized by using a phosphorus dissolving capacity determination culture medium comprising 10g of glucose, 5g of an insoluble phosphorus source and MgCl25g，(NH4)2SO40.1g，KCl 0.2g，MgSO40.25g, 1000ml of distilled water, 20g of agar in a solid medium, and a natural pH.
Further, the indissolvable state phosphorus source is tricalcium phosphate or ground phosphate rock or lecithin.
The invention also provides application of the bacillus megaterium QZY-3 in promoting crop growth.
The invention also provides a preparation for promoting the growth of crops, which comprises the bacillus megaterium QZY-3 or strain fermentation liquor.
The invention also provides application of the bacillus megaterium QZY-3 in preparation of organic fertilizer additives and bio-organic fertilizers.
Compared with the prior art, the invention has the following beneficial effects:
the bacillus megaterium QZY-3 does not need to be tested in the aspect of chemical toxicology, and belongs to safe strains; the strain has obvious dissolving capacity to inorganic insoluble phosphorus (tricalcium phosphate and ground phosphate rock) and organic insoluble phosphorus (lecithin), and has extremely obvious growth promoting effect on wheat seedlings and rape seedlings. Has good application prospect in the aspects of researching and developing microbial growth promoting inoculants, microbial fertilizers, popularization and demonstration on soil and the like.
Drawings
FIG. 1 is a microscopic examination picture of Bacillus megaterium QZY-3 cultured on beef extract peptone medium at a constant temperature of 32 ℃ for 24 hours.
FIG. 2 shows the colony morphology of Bacillus megaterium QZY-3 on a plate.
FIG. 3 is a phylogenetic tree of Bacillus megaterium QZY-3 of the present invention.
FIG. 4 is one of the comparative plots for growth of wheat seedlings in example 3.
FIG. 5 is a second graph comparing the growth of wheat seedlings in example 3.
FIG. 6 is one of the comparative plots for the growth of seedlings of Brassica napus in example 3.
FIG. 7 is a second graph showing the comparison between the growth of seedlings of Brassica napus in example 3.
Detailed Description
The present invention is further illustrated by the following specific examples.
Example 1
Isolation and identification of strains
The strain is obtained by separating and screening yellow soil farmland in Guizhou, and the morphological and molecular biological identification of the strain is as follows:
(1) morphological and physiological and biochemical characteristics of the strain
The strain is cultured for 24 hours at a constant temperature of 32 ℃ on a beef extract peptone culture medium (3 g of beef extract, 10g of peptone, 5g of NaCl, 15g of agar and 1000ml of distilled water, wherein the pH value of a culture solution is 7.0-7.2) consisting of the following components in parts by mass/volume, as shown in figure 1, the strain forms a large number of spores, forms long rods (1.2-1.5 multiplied by 2.8-4.0 mu m) of the pre-spore bacteria, and is arranged into single or chain; gram-positive; after 3 days of culture, as shown in FIG. 2, the colony morphology was: the bacterial colony is small and round, milky yellow, neat in edge, smooth and moist in surface and non-transparent; the puncture inoculation cultured thalli grows in a cloud-like diffusion mode in a culture medium of a test tube, and the motility is shown; the characteristics of the product are shown in table 1.
TABLE 1 physiological and biochemical characteristics of Bacillus megaterium QZY-3
| Detecting the index | Results | Detecting the index | Results |
| Nitrate reduction | + | Citrate utilization | + |
| Contact enzyme | + | Phenylalanine deaminase | + |
| V-P assay | — | Producing indole | + |
| Methyl red (M.R) | + | Lecithinase | + |
| Starch hydrolysis | + | Oxidative fermentation of glucose | + |
| Salt tolerance and salt demand | Liquefaction of gelatin | + | |
| 2％NaCl | + | Hydrolysis of tyrosine | + |
| 5％NaCl | + | Oxidase enzyme | + |
| 7％NaCl | + | Anaerobic growth | — |
| 10％NaCl | — | Growth in the presence of lysozyme | — |
Note: "+" indicates that the indicator is positive; "-" indicates that the index is negative.
(2) 16S rDNA sequence of strain and analysis
The length of the 16S rDNA gene sequence of the strain is 1485bp, and the sequence is as follows:
the 16S rDNA sequence of the Bacillus megaterium QZY-3 is input into a GeneBank database of NCBI and is compared with the 16S rDNA gene sequence of a known strain, so that the 16S rDNA sequence of the Bacillus megaterium QZY-3 has the highest homology with the Bacillus megaterium, which reaches 100 percent and is shown in Table 2.
TABLE 2 homology alignment of B.megaterium QZY-3
The phylogenetic tree of Bacillus megaterium QZY-3 of the present invention is shown in FIG. 3.
Example 2
Determination of phosphorus dissolving capacity of Bacillus megaterium QZY-3
1. A culture medium; (1) bacillus megaterium QZY-3 phosphorus solubilizing ability determination medium (NBRIP medium): 10g of glucose, 5g of insoluble phosphorus source (tricalcium phosphate or ground phosphate rock or lecithin), and MgCl25g，(NH4)2SO40.1g，KCl 0.2g，MgSO40.25g, 1000ml of distilled water, 20g of agar added into a solid culture medium, and natural pH; (2) b, B.megaterium QZY-3 preservation and fermentation medium: 3.0g of beef extract, 10.0g of peptone, 5.0g of NaCl and 1000ml of distilled water, adding 15g of agar into a solid culture medium, and adjusting the pH to 7.0-7.2.
2. The method for measuring the phosphorus dissolving capacity comprises the following steps: inoculating the screened bacillus megaterium QZY-3 into a sterilized culture solution for fermentation, and culturing for 24 hours at the temperature of 2 ℃ and the speed of 160r/min to obtain a fermentation liquid; inoculating bacillus megaterium QZY-3 fermentation liquor into a sterilized NBRIP liquid culture medium according to the inoculation amount of 1%, carrying out shake culture at 28 ℃ at 150r/min, centrifuging the culture solution for culturing the 7 th day at 4 ℃ at 6000r/min for 5-8min, taking 1ml of supernatant, measuring the effective phosphorus content by a molybdenum-antimony anti-colorimetric method, repeating the steps for three times, and simultaneously setting a blank control of non-inoculated bacteria, wherein the phosphorus dissolving capacity of the strain is the difference value of the effective phosphorus content of the inoculated culture solution and the blank control. The results of the measurement of the phosphorus-solubilizing ability are shown in Table 3.
TABLE 3 solubility of Bacillus megaterium QZY-3 in insoluble phosphates (tricalcium phosphate, ground phosphate rock, lecithin)
As can be seen from Table 3, the Bacillus megaterium QZY-3 has strong phosphorus dissolving capacity and can be used for preparing organic fertilizer additives and biological organic fertilizers.
Example 3
Growth test of Bacillus megaterium QZY-3 for promoting crop seedlings (rape and wheat)
1. Preparing fermentation liquor: the bacillus megaterium QZY-3 is inoculated into a sterilized culture solution for fermentation, the culture is carried out for 24 hours at 32 ℃ and 160r/min, fermentation liquor is obtained, the fermentation liquor and sterile water are diluted for later use according to the proportion of 1:10, meanwhile, the sterilized culture solution without inoculation is set as a contrast, and the dilution is carried out according to the proportion of 1: 10.
2. The rape growth promotion test method comprises the following steps: taking 9 culture dish plates with the diameter of 90mm, paving a thin paper towel on each plate, wetting the culture dish plates with sterile water, placing 30 rape seeds on each plate, placing the plate under the condition of 28 ℃ for culturing and uninterruptedly supplementing the sterile water until the seeds germinate, removing the non-germinated seeds, averagely dividing the 9 plates into three groups, adding 10ml of diluted bacillus megaterium QZY-3 fermentation liquor into each plate of the first group, adding 10ml of sterile water into each plate of the second group, adding 10ml of diluted sterile culture solution into each plate of the third group, continuing placing the plate under the condition of 28 ℃ for culturing and uninterruptedly supplementing the sterile water, observing the growth condition after 15 days, randomly extracting 5 seedlings from each plate and measuring the length of the seedlings, wherein the result is shown in figure 6, figure 7 and table 4.
3. The wheat growth promotion test method comprises the following steps: taking 9 culture dish plates with the diameter of 90mm, paving a thin paper towel on each plate, wetting with sterile water, placing 10 rape seeds on each plate, placing the plates at 28 ℃ for culture and continuously supplementing sterile water until germination, removing non-germinated seeds, averagely dividing the 9 plates into three groups, adding 10ml of diluted QZY-3 fermentation liquor into each plate of the first group, adding 10ml of sterile water into each plate of the second group, adding 10ml of diluted sterile culture liquor into each plate of the third group, continuously placing the plates at 28 ℃ for culture and continuously supplementing sterile water, observing the growth condition after 15 days, randomly extracting 5 seedlings from each plate and measuring the length, and obtaining the result shown in figure 4, figure 5 and table 5.
TABLE 4 growth promotion test results of Bacillus megaterium QZY-3 on rape seedlings
Note that different lower case letters in the same column in the table indicate significant differences at the 0.05 level (i.e., p <0.05), as follows
As can be seen from FIG. 6, FIG. 7 and Table 4, Bacillus megaterium QZY-3 of the present invention has a significant effect on the growth promotion of oilseed rape seedlings.
TABLE 5 growth promotion test results of Bacillus megaterium QZY-3 on wheat seedlings
As can be seen from FIG. 4, FIG. 5 and Table 5, Bacillus megaterium QZY-3 of the present invention has a significant effect on promoting the growth of wheat seedlings. In conclusion, the bacillus megaterium QZY-3 can be used for preparing preparations for promoting the growth of crops.
<110> Qiaoqinhei great
<120> bacillus megaterium QZY-3 and application thereof
<160>1
<210>1
<211>1485
<212>DNA
<213> Bacillus megaterium QZY-3(Bacillus megaterium QZY-3)
<220>
<223> 16S rDNA of Bacillus megaterium QZY-3
<400>1
GGCTCAGGAT GAACGCTGGC GGCGTGCCTA ATACATGCAA GTCGAGCGAA CTGATTAGAA 60
GCTTGCTTCT ATGACGTTAG CGGCGGACGG GTGAGTAACA CGTGGGCAAC CTGCCTGTAA 120
GACTGGGATA ACTTCGGGAA ACCGAAGCTA ATACCGGATA GGATCTTCTC CTTCATGGGA 180
GATGATTGAA AGATGGTTTC GGCTATCACT TACAGATGGG CCCGCGGTGC ATTAGCTAGT 240
TGGTGAGGTA ACGGCTCACC AAGGCAACGA TGCATAGCCG ACCTGAGAGG GTGATCGGCC 300
ACACTGGGAC TGAGACACGG CCCAGACTCC TACGGGAGGC AGCAGTAGGG AATCTTCCGC 360
AATGGACGAA AGTCTGACGG AGCAACGCCG CGTGAGTGAT GAAGGCTTTC GGGTCGTAAA 420
ACTCTGTTGT TAGGGAAGAA CAAGTACGAG AGTAACTGCT CGTACCTTGA CGGTACCTAA 480
CCAGAAAGCC ACGGCTAACT ACGTGCCAGC AGCCGCGGTA ATACGTAGGT GGCAAGCGTT 540
ATCCGGAATT ATTGGGCGTA AAGCGCGCGC AGGCGGTTTC TTAAGTCTGA TGTGAAAGCC 600
CACGGCTCAA CCGTGGAGGG TCATTGGAAA CTGGGGAACT TGAGTGCAGA AGAGAAAAGC 660
GGAATTCCAC GTGTAGCGGT GAAATGCGTA GAGATGTGGA GGAACACCAG TGGCGAAGGC 720
GGCTTTTTGG TCTGTAACTG ACGCTGAGGC GCGAAAGCGT GGGGAGCAAA CAGGATTAGA 780
TACCCTGGTA GTCCACGCCG TAAACGATGA GTGCTAAGTG TTAGAGGGTT TCCGCCCTTT 840
AGTGCTGCAG CTAACGCATT AAGCACTCCG CCTGGGGAGT ACGGTCGCAA GACTGAAACT 900
CAAAGGAATT GACGGGGGCC CGCACAAGCG GTGGAGCATG TGGTTTAATT CGAAGCAACG 960
CGAAGAACCT TACCAGGTCT TGACATCCTC TGACAACTCT AGAGATAGAG CGTTCCCCTT 1020
CGGGGGACAG AGTGACAGGT GGTGCATGGT TGTCGTCAGC TCGTGTCGTG AGATGTTGGG 1080
TTAAGTCCCG CAACGAGCGC AACCCTTGAT CTTAGTTGCC AGCATTTAGT TGGGCACTCT 1140
AAGGTGACTG CCGGTGACAA ACCGGAGGAA GGTGGGGATG ACGTCAAATC ATCATGCCCC 1200
TTATGACCTG GGCTACACAC GTGCTACAAT GGATGGTACA AAGGGCTGCA AGACCGCGAG 1260
GTCAAGCCAA TCCCATAAAA CCATTCTCAG TTCGGATTGT AGGCTGCAAC TCGCCTACAT 1320
GAAGCTGGAA TCGCTAGTAA TCGCGGATCA GCATGCCGCG GTGAATACGT TCCCGGGCCT 1380
TGTACACACC GCCCGTCACA CCACGAGAGT TTGTAACACC CGAAGTCGGT GGAGTAACCG 1440
TAAGGAGCTA GCCGCCTAAG GTGGGACAGA TGATTGGGTG AAGTC 1485
Claims (8)
1. The bacillus megaterium QZY-3 is characterized by being preserved in China center for type culture Collection with the preservation number of CCTCC NO: m2019717.
2. The preparation method of the strain fermentation liquor is characterized by comprising the following steps: taking a preservation number of CCTCC NO: inoculating the Bacillus megaterium QZY-3 of M2019717 into a sterilized culture medium for fermentation, and culturing at 32 ℃ and 160r/min for 24 hours to obtain a fermentation liquid; wherein the storage and fermentation culture medium of the bacillus megaterium QZY-3 comprises 3.0g of beef extract, 10.0g of peptone, 5.0g of NaCl and 1000ml of distilled water, 15g of agar is added into a solid culture medium, and the pH is adjusted to 7.0-7.2.
3. Use of the Bacillus megaterium QZY-3 of claim 1 for solubilizing phosphorus in a sparingly soluble state.
4. The macro of claim 3The application of the bacillus megaterium QZY-3 in dissolving insoluble phosphorus is characterized in that the used phosphorus dissolving capacity determination culture medium comprises 10g of glucose, 5g of insoluble phosphorus source and MgCl25g，(NH4)2SO40.1g， KCl 0.2g，MgSO40.25g, 1000ml of distilled water, 20g of agar in a solid medium, and a natural pH.
5. The use of Bacillus megaterium QZY-3 in solubilizing phosphorus in an insoluble form as claimed in claim 4, wherein the source of phosphorus in an insoluble form is tricalcium phosphate or ground phosphate rock or lecithin.
6. Use of bacillus megaterium QZY-3 according to claim 1 for promoting the growth of crops.
7. A crop growth promoting agent comprising Bacillus megaterium QZY-3 of claim 1 or the fermentation broth of the strain of claim 4.
8. The use of Bacillus megaterium QZY-3 of claim 1 in the preparation of organic fertilizer additives and bio-organic fertilizers.
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