CN118165892B - Pseudomonas composite microbial agent and application thereof - Google Patents
Pseudomonas composite microbial agent and application thereof Download PDFInfo
- Publication number
- CN118165892B CN118165892B CN202410570976.6A CN202410570976A CN118165892B CN 118165892 B CN118165892 B CN 118165892B CN 202410570976 A CN202410570976 A CN 202410570976A CN 118165892 B CN118165892 B CN 118165892B
- Authority
- CN
- China
- Prior art keywords
- pseudomonas
- fermentation
- culture medium
- microbial inoculum
- liquid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241000589516 Pseudomonas Species 0.000 title claims abstract description 150
- 239000002131 composite material Substances 0.000 title claims abstract description 40
- 230000000813 microbial effect Effects 0.000 title claims description 39
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 71
- 239000002689 soil Substances 0.000 claims abstract description 46
- 238000004321 preservation Methods 0.000 claims abstract description 43
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 31
- 230000012010 growth Effects 0.000 claims abstract description 18
- 230000001737 promoting effect Effects 0.000 claims abstract description 9
- 238000000855 fermentation Methods 0.000 claims description 92
- 230000004151 fermentation Effects 0.000 claims description 92
- 239000007788 liquid Substances 0.000 claims description 39
- 239000001963 growth medium Substances 0.000 claims description 38
- 239000003795 chemical substances by application Substances 0.000 claims description 37
- 238000009630 liquid culture Methods 0.000 claims description 34
- 239000006228 supernatant Substances 0.000 claims description 25
- 239000012880 LB liquid culture medium Substances 0.000 claims description 19
- 241001052560 Thallis Species 0.000 claims description 18
- 239000008223 sterile water Substances 0.000 claims description 18
- 230000001580 bacterial effect Effects 0.000 claims description 16
- 238000009629 microbiological culture Methods 0.000 claims description 16
- 239000011248 coating agent Substances 0.000 claims description 11
- 238000000576 coating method Methods 0.000 claims description 11
- 238000002156 mixing Methods 0.000 claims description 11
- 241000196324 Embryophyta Species 0.000 claims description 9
- 238000002360 preparation method Methods 0.000 claims description 8
- 235000006008 Brassica napus var napus Nutrition 0.000 claims description 5
- 241000589774 Pseudomonas sp. Species 0.000 claims description 5
- 244000005700 microbiome Species 0.000 claims description 5
- 235000014698 Brassica juncea var multisecta Nutrition 0.000 claims description 3
- 240000000385 Brassica napus var. napus Species 0.000 claims description 3
- 235000006618 Brassica rapa subsp oleifera Nutrition 0.000 claims description 3
- 235000004977 Brassica sinapistrum Nutrition 0.000 claims description 3
- 230000002378 acidificating effect Effects 0.000 claims description 2
- 239000002253 acid Substances 0.000 abstract description 28
- 238000011282 treatment Methods 0.000 abstract description 17
- 150000001875 compounds Chemical class 0.000 abstract description 8
- 239000002609 medium Substances 0.000 description 18
- 230000000694 effects Effects 0.000 description 15
- 239000004033 plastic Substances 0.000 description 14
- 238000012258 culturing Methods 0.000 description 12
- 241000894006 Bacteria Species 0.000 description 11
- 239000007787 solid Substances 0.000 description 11
- 238000012360 testing method Methods 0.000 description 11
- 239000000919 ceramic Substances 0.000 description 7
- 108020004465 16S ribosomal RNA Proteins 0.000 description 6
- 240000002791 Brassica napus Species 0.000 description 6
- 229930002875 chlorophyll Natural products 0.000 description 6
- 235000019804 chlorophyll Nutrition 0.000 description 6
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 6
- 230000006872 improvement Effects 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- 238000012216 screening Methods 0.000 description 5
- 239000000758 substrate Substances 0.000 description 5
- 229920001817 Agar Polymers 0.000 description 4
- 235000011293 Brassica napus Nutrition 0.000 description 4
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 239000008367 deionised water Substances 0.000 description 4
- 229910021641 deionized water Inorganic materials 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 238000002791 soaking Methods 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 238000011161 development Methods 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 239000012137 tryptone Substances 0.000 description 3
- PWVXXGRKLHYWKM-UHFFFAOYSA-N 5-[2-(benzenesulfonyl)ethyl]-3-[(1-methylpyrrolidin-2-yl)methyl]-1h-indole Chemical compound CN1CCCC1CC(C1=C2)=CNC1=CC=C2CCS(=O)(=O)C1=CC=CC=C1 PWVXXGRKLHYWKM-UHFFFAOYSA-N 0.000 description 2
- 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 description 2
- 235000010469 Glycine max Nutrition 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 239000006004 Quartz sand Substances 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 229910052564 epsomite Inorganic materials 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 229910052603 melanterite Inorganic materials 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000005507 spraying Methods 0.000 description 2
- 239000013589 supplement Substances 0.000 description 2
- 230000001502 supplementing effect Effects 0.000 description 2
- 229960004799 tryptophan Drugs 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 235000011331 Brassica Nutrition 0.000 description 1
- 241000219198 Brassica Species 0.000 description 1
- 241000219193 Brassicaceae Species 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 1
- 240000008790 Musa x paradisiaca Species 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 241001312486 Pseudomonas migulae Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 240000003768 Solanum lycopersicum Species 0.000 description 1
- 230000006518 acidic stress Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- DLHONNLASJQAHX-UHFFFAOYSA-N aluminum;potassium;oxygen(2-);silicon(4+) Chemical compound [O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[Al+3].[Si+4].[Si+4].[Si+4].[K+] DLHONNLASJQAHX-UHFFFAOYSA-N 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000021015 bananas Nutrition 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 239000011449 brick Substances 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000008157 edible vegetable oil Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- -1 i.e. Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 239000010815 organic waste Substances 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- IFGCUJZIWBUILZ-UHFFFAOYSA-N sodium 2-[[2-[[hydroxy-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyphosphoryl]amino]-4-methylpentanoyl]amino]-3-(1H-indol-3-yl)propanoic acid Chemical compound [Na+].C=1NC2=CC=CC=C2C=1CC(C(O)=O)NC(=O)C(CC(C)C)NP(O)(=O)OC1OC(C)C(O)C(O)C1O IFGCUJZIWBUILZ-UHFFFAOYSA-N 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a pseudomonas composite microbial inoculum, which comprises pseudomonas N134 and pseudomonas N144, wherein the preservation number of the pseudomonas N134 is CGMCC NO.29191; the preservation number of the pseudomonas N144 is CGMCC NO.29192. The invention also discloses application of the composite microbial inoculum in promoting the growth of acid soil rape. The screened pseudomonas N134 and pseudomonas N144 are prepared into the compound microbial inoculum, and compared with the treatment of clear water, each single microbial inoculum and other compound microbial inoculum, the compound microbial inoculum has a more promoting effect on the growth of rape in acid soil, and provides support for improving the productivity of the acid soil.
Description
Technical Field
The invention relates to the technical field of agricultural microorganisms, in particular to a pseudomonas composite microbial agent and application thereof.
Background
The acid soil is a soil general term with pH less than 7, and the acid soil accounts for about 40% of the cultivated land soil worldwide at present and comprises brick red soil, yellow soil, dry red soil and other soil types. In tropical and subtropical areas in south China, acid soil such as red soil, yellow soil and red yellow soil is widely distributed, the acid soil is distributed in 15 provinces (areas), and the acid soil mainly occupies 22.7% of the total area of the whole national land in Jiangxi provinces, hunan provinces (areas) and the like respectively, and the acid soil area shows an increasing trend. Under acid stress, the microbial population in the soil is obviously reduced, the growth of plants is limited, the yield is reduced, and the vegetation health in agriculture and ecosystems is affected. Therefore, the improvement and repair of the acid soil are of great significance to the protection of ecological environment and the guarantee of sustainable development of agriculture. At present, most of the acid soil improvement adopts physical improvement and chemical improvement, the prior art has little research on microbial improvement, and strain resources applicable to actual production are still relatively deficient.
The microbial agent is divided into a single microbial agent and a composite microbial agent, wherein the single microbial agent is prepared from a functional microorganism; the composite microbial agent consists of two or more beneficial microorganisms. The single microbial inoculum has single function and unstable effect, and cannot meet the ecological stability of multiple functions and complex rhizosphere environments. The compound microbial inoculum has synergistic effect, can solve the problems of single microbial inoculum action mechanism, unstable effect and the like, and the compound bacterial strain can influence the physiological metabolism activity of plants by secreting different physiological active substances to promote the growth of the plants, thereby having the advantages of more variety, complete functions and better promotion effect.
Rape is a one-year or two-year-old herb plant of cruciferae and Brassica, has strong winter, is often utilized by human beings in the form of edible oil or vegetables, has important value in economy, ecology and ornamental aspects, and is one of main economic crops planted in southern acid soil dry lands in China. Therefore, rape is used as a test crop in the invention.
Disclosure of Invention
In order to overcome the problems, the invention provides a pseudomonas composite microbial inoculum and application thereof.
The aim of the invention can be achieved by the following technical scheme:
The invention provides a Pseudomonas composite microbial agent, which comprises Pseudomonas N134 and Pseudomonas N144, wherein the classification of the Pseudomonas N134 is named as Pseudomonas N134, and the Pseudomonas N134 is preserved in China general microbiological culture Collection center (China general microbiological culture Collection center) with the preservation date of 2023 and 11 months and 30 days, and the preservation number of CGMCC NO.29191; the classification of the Pseudomonas N144 is named as Pseudomonas N144, and the Pseudomonas N144 is preserved in China general microbiological culture Collection center (CGMCC) with the preservation date of 2023, 11 and 30 and the preservation number of CGMCC NO.29192.
Further, the composite microbial inoculum is prepared by the following steps:
(1) Respectively carrying out liquid fermentation on pseudomonas N134 with the preservation number of CGMCC No.29191 and pseudomonas N144 with the preservation number of CGMCC No.29192 to obtain pseudomonas N134 fermentation liquor and pseudomonas N144 fermentation liquor;
(2) Centrifuging the pseudomonas N134 fermentation liquor and the pseudomonas N144 fermentation liquor respectively, removing supernatant, and re-suspending with sterile water to obtain a pseudomonas N134 microbial agent and a pseudomonas N144 microbial agent;
(3) And mixing the pseudomonas N134 microbial inoculum and the pseudomonas N144 microbial inoculum in equal volumes to obtain the composite microbial inoculum.
Further, the composite microbial inoculum is prepared by the following steps:
(1) Inoculating pseudomonas N134 with the preservation number of CGMCC NO.29191 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium comprises a TSB liquid culture medium, an LB liquid culture medium or an R2A liquid culture medium, and the liquid fermentation conditions are as follows: the temperature is 28-30 ℃, the rotating speed is 170-200rpm, the fermentation time is 1-2d, the pseudomonas N134 fermentation broth is obtained, the number of the coating and counting thalli is diluted, and the concentration of the thalli of the pseudomonas N134 fermentation broth is more than or equal to 1 multiplied by 10 8 cfu/mL;
Inoculating pseudomonas N144 with the preservation number of CGMCC NO.29192 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium comprises a TSB liquid culture medium, an LB liquid culture medium or an R2A liquid culture medium, and the liquid fermentation conditions are as follows: the temperature is 28-30 ℃, the rotating speed is 170-200rpm, the fermentation time is 1-2d, the pseudomonas N144 fermentation broth is obtained, the number of the coating and counting thalli is diluted, and the concentration of the thalli of the pseudomonas N144 fermentation broth is more than or equal to 1 multiplied by 10 8 cfu/mL;
(2) Centrifuging the pseudomonas N134 fermentation liquor, removing the supernatant, and re-suspending the supernatant by using sterile water with ten times the volume of the pseudomonas N134 fermentation liquor to obtain a pseudomonas N134 microbial inoculum;
centrifuging the pseudomonas N144 fermentation liquor, removing the supernatant, and re-suspending the supernatant by using sterile water with ten times the volume of the pseudomonas N144 fermentation liquor to obtain a pseudomonas N144 microbial inoculum;
(3) And mixing the pseudomonas N134 microbial inoculum and the pseudomonas N144 microbial inoculum in equal volumes to obtain the composite microbial inoculum.
The second aspect of the invention provides a preparation method of a Pseudomonas composite microbial agent, wherein the composite microbial agent comprises Pseudomonas N134 and Pseudomonas N144, the classification of the Pseudomonas N134 is named as Pseudomonas N134, and the Pseudomonas N134 is preserved in China general microbiological culture Collection center (China general microbiological culture Collection center) with the preservation date of 2023, 11 months and 30 days and the preservation number of CGMCC NO.29191; the classification of the Pseudomonas N144 is named as Pseudomonas N144, and the Pseudomonas N144 is preserved in the China general microbiological culture Collection center (China Committee) for culture Collection of microorganisms, wherein the preservation date is 2023, 11 and 30, and the preservation number is CGMCC NO.29192;
The preparation method of the composite microbial inoculum comprises the following steps:
(1) Respectively carrying out liquid fermentation on pseudomonas N134 with the preservation number of CGMCC No.29191 and pseudomonas N144 with the preservation number of CGMCC No.29192 to obtain pseudomonas N134 fermentation liquor and pseudomonas N144 fermentation liquor;
(2) Centrifuging the pseudomonas N134 fermentation liquor and the pseudomonas N144 fermentation liquor respectively, removing supernatant, and re-suspending with sterile water to obtain a pseudomonas N134 microbial agent and a pseudomonas N144 microbial agent;
(3) And mixing the pseudomonas N134 microbial inoculum and the pseudomonas N144 microbial inoculum in equal volumes to obtain the composite microbial inoculum.
Further, the composite microbial inoculum is prepared by the following steps:
(1) Inoculating pseudomonas N134 with the preservation number of CGMCC NO.29191 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium comprises a TSB liquid culture medium, an LB liquid culture medium or an R2A liquid culture medium, and the liquid fermentation conditions are as follows: the temperature is 28-30 ℃, the rotating speed is 170-200rpm, the fermentation time is 1-2d, the pseudomonas N134 fermentation broth is obtained, the number of the coating and counting thalli is diluted, and the concentration of the thalli of the pseudomonas N134 fermentation broth is more than or equal to 1 multiplied by 10 8 cfu/mL;
Inoculating pseudomonas N144 with the preservation number of CGMCC NO.29192 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium comprises a TSB liquid culture medium, an LB liquid culture medium or an R2A liquid culture medium, and the liquid fermentation conditions are as follows: the temperature is 28-30 ℃, the rotating speed is 170-200rpm, the fermentation time is 1-2d, the pseudomonas N144 fermentation broth is obtained, the number of the coating and counting thalli is diluted, and the concentration of the thalli of the pseudomonas N144 fermentation broth is more than or equal to 1 multiplied by 10 8 cfu/mL;
(2) Centrifuging the pseudomonas N134 fermentation liquor, removing the supernatant, and re-suspending the supernatant by using sterile water with ten times the volume of the pseudomonas N134 fermentation liquor to obtain a pseudomonas N134 microbial inoculum;
centrifuging the pseudomonas N144 fermentation liquor, removing the supernatant, and re-suspending the supernatant by using sterile water with ten times the volume of the pseudomonas N144 fermentation liquor to obtain a pseudomonas N144 microbial inoculum;
(3) And mixing the pseudomonas N134 microbial inoculum and the pseudomonas N144 microbial inoculum in equal volumes to obtain the composite microbial inoculum.
The third aspect of the invention provides application of the pseudomonas composite microbial inoculum in promoting the growth of acid soil rape.
Further, the pH of the acid soil is 4.5-6.5.
Further, in the application, the microbial inoculum is applied for a period of time after the rape seedlings are transplanted, and the application amount is 30-50 mL/rape seedling.
Furthermore, in the application process, the microbial inoculum is applied after the rape seedlings are transplanted for 7-10d, and the application amount is 30-50 mL/rape seedling.
The invention has the beneficial effects that:
The screened pseudomonas N134 and pseudomonas N144 are prepared into the compound microbial inoculum, and compared with the treatment of clear water, each single microbial inoculum and other compound microbial inoculum, the compound microbial inoculum has a more promoting effect on the growth of rape in acid soil, and provides support for improving the productivity of the acid soil.
Drawings
FIG. 1 is a photograph of a plate of strain N134.
FIG. 2 is a photograph of a flat panel of strain 144.
FIG. 3 is a 16S rRNA phylogenetic tree of strain N134.
FIG. 4 is a 16S rRNA phylogenetic tree of strain N144.
FIG. 5 is a graph of the phosphorus-solubilizing effect of the strain (including N134 and N111).
FIG. 6 is a graph of the phosphorus-solubilizing effect of the strain (including N144).
FIG. 7 shows the effect of treatment with different single strains on the fresh weight of greenhouse canola.
FIG. 8 shows the effect of treatments with different single strains on the dry weight of greenhouse oilseed rape.
FIG. 9 is the effect of treatment with different single strains on the chlorophyll content (SPAD) of greenhouse rape.
FIG. 10 shows the affinity (i.e., coating opposition) of the strains.
FIG. 11 shows the effect of treatment with different strains on the fresh weight of greenhouse canola.
FIG. 12 shows the effect of treatments with different strains on the dry weight of greenhouse oilseed rape.
FIG. 13 is the effect of treatment with different strains on the chlorophyll content (SPAD) of greenhouse rape.
The different letters above the histogram are significant differences between the different treatments (p < 0.05).
Biological material preservation information
N134, classified and named as Pseudomonas sp, is preserved in China general microbiological culture Collection center (China Committee for culture Collection), and has a preservation address of North Star paths 1 to the Yangzhang area of Beijing, 3 rd national academy of sciences of China, a preservation date of 2023, 11 months and 30 days, and a preservation number of CGMCC No.29191.
N144, classified and named as Pseudomonas sp, is preserved in China general microbiological culture Collection center (CGMCC) with a preservation address of North Star in the Guangdong national institute of sciences of China, no. 3, beijing city, a preservation date of 2023, 11 months and 30 days, and a preservation number of CGMCC No.29192.
Detailed Description
The following examples facilitate a better understanding of the present invention, but are not to be construed as limiting the same. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the examples described below, unless otherwise specified, were purchased from conventional biochemical reagent stores.
Unless otherwise specified, the raw materials used in the following examples are as follows:
The seedling raising substrate is a special substrate for raising seedlings of Xingxing-nong cards produced by Jiangsu Xingjing substrate technology Co., ltd, and the product number is: 161102G0097N.
TSB solid medium, i.e., tryptone soy broth agar medium (1L): 30g of tryptone soybean broth, deionized water to a volume of 1L,20g of agar powder, and sterilizing at 115 ℃ for 30min.
LB liquid Medium (1L): 10g of tryptone, 5g of yeast powder and 10g of NaCl, and sterilizing the mixture for 30min at 115 ℃ with deionized water to reach a volume of 1L.
Phosphate-dissolving solid medium (1L): glucose 10g,(NH4)2SO4 0.5g,NaCl 0.3g,KCl 0.3g,FeSO4·7H2O 0.3g,MgSO4·7H2O 0.3g,MnSO4·4H2O 0.03g, yeast extract 0.4g, ca 3(PO4)2 g, deionized water to 1L, agar powder 20g, sterilizing at 115 ℃ for 30min.
Potassium-dissolving solid culture medium (1L): glucose 10g,NH4NO3 3.0g,NaH2PO4 1g,FeSO4·7H2O 1g,MgSO4·7H2O 1g, potassium feldspar powder 1.5g, deionized water to 1L, agar powder 20g, and sterilizing at 115 ℃ for 30min.
EXAMPLE 1 selection of acid-resistant strains
The first step: shaking bacteria
202 Strains of bacteria separated from soil or rhizosphere for planting crops such as bananas, tomatoes and the like, which are saved in a key laboratory of high-technology research on recycling of solid organic wastes in Jiangsu province, are activated, and are inoculated into 3mL of LB liquid medium and cultured for 2d at 30 ℃ and 170rpm, so that fermentation liquor is obtained.
And a second step of: determination of growth Properties of Strain under acidic conditions
① Centrifuging the fermentation broth cultured in the first step at 4000rpm for 6min, removing supernatant, re-suspending with sterile water to obtain bacterial suspension, and regulating the OD600 of the bacterial suspension to 1 with sterile water.
② Preparing an LB liquid culture medium, regulating the pH value of the LB liquid culture medium to 4.0 and 5.0 respectively by using 0.04mol/L dilute hydrochloric acid, and setting the LB liquid culture medium (pH=6.86) with the pH value not regulated as a control; respectively adding the prepared LB liquid culture media with different pH values into three 96-well plates for standby according to 180 mu L of each well, sequentially inoculating bacterial suspension diluted in 20 mu L ① into the LB liquid culture media with different pH values, setting 4 times of repetition for each strain, placing the strain in a constant-temperature incubator at 30 ℃ for culturing for 1d and 2d, respectively measuring OD 600 of each strain after culturing for 1d and 2d by using an enzyme-labeling instrument, wherein higher OD value indicates stronger growth performance of the strain; the 202 strains to be tested can grow in LB liquid medium without regulating the pH value, 178 strains can grow in LB liquid medium with the pH value of 5.0, 9 strains can grow in LB liquid medium with the pH value of 4.0 (although the 9 strains can grow in LB liquid medium with the pH value of 4.0, the 9 strains are identified to have no function, and the subsequent experiment does not involve the 9 strains). The strains N134 and N144 can grow under the condition that the pH is 5.0.
Example 2 determination of Strain functionality
The acid-resistant strain of 178 strains selected in example 1, which was able to grow in LB liquid medium at pH 5.0, was further assayed for its functionality.
(1) Screening of acid-tolerant IAA-producing strains
The pH of the LB liquid medium was adjusted to 5.0 with 0.04mol/L dilute hydrochloric acid, L-tryptophan (0.1 mg/mL) was added, the prepared LB liquid medium containing L-tryptophan was packed into shaking tubes, 3mL of each tube was used, and the acid-resistant strain selected in example 1 was inoculated into the LB liquid medium containing L-tryptophan, and after shaking culture at 30℃and 170rpm for 2d, a bacterial liquid was obtained.
And (3) dripping 100 mu L of bacteria on a white ceramic plate, simultaneously adding an equal volume of colorimetric solution (1 g of Fecl 3·6H2 O is weighed and dissolved in 21.485mL of concentrated H 2SO4, the solution slowly falls into distilled water to be diluted, and then the distilled water is used for fixing the volume to 50mL to obtain the colorimetric solution), and taking a mixed solution of 100 mu L of non-inoculated LB liquid culture medium (the pH value is adjusted to 5.0, the method is the same as above) and the equal volume of colorimetric solution as a control. After the white ceramic plate is placed at room temperature and is kept away from light for 30min, the white ceramic plate is observed, the color of the white ceramic plate turns pink, the color of the white ceramic plate is deeper, the secretion strength of the white ceramic plate is higher, and the white ceramic plate cannot secrete IAA.
The strain with IAA secretion capability obtained by the primary screening is quantitatively measured, 1mL of bacterial liquid is sucked and centrifuged at 1000rpm for 10min, supernatant is taken, an equal volume of colorimetric liquid is added, the mixture is kept stand for 30min in dark place, 3 repeats are arranged for each strain, and OD 530 value is measured. And (5) calculating the IAA content in the unit volume of bacterial liquid according to a standard curve.
(2) Screening of phosphorus-dissolving strains
Preparing an LB liquid culture medium, subpackaging the LB liquid culture medium into shaking tubes, and inoculating 3mL of acid-resistant strains selected in the example 1 into the LB liquid culture medium respectively, and performing shaking culture at 30 ℃ and 170rpm for 2d to obtain bacterial liquid. The bacterial liquid is coated on a phosphorus-dissolving solid culture medium flat plate, and the flat plate is placed in a 30 ℃ incubator for 3-5d. Colonies with phosphorus-dissolving rings and different forms are selected.
(3) Screening of Potassium-decomposing Strain
Preparing an LB liquid culture medium, subpackaging the LB liquid culture medium into shaking tubes, and inoculating 3mL of acid-resistant strains selected in the example 1 into the LB liquid culture medium respectively, and performing shaking culture at 30 ℃ and 170rpm for 2d to obtain bacterial liquid. The bacterial liquid is coated on a potassium-dissolving solid culture medium plate, and the plate is placed in a 30 ℃ incubator for 3-5d. During the culture, the colony morphology is observed, and the colony is round, transparent, high in bulge, moist, sticky and elastic.
Finally, respectively obtaining 22 strains of acid-resistant IAA-producing functional bacteria, 6 strains of phosphorus-dissolving functional bacteria and 7 strains of potassium-dissolving functional bacteria, respectively screening out 3 strains with better functional effects from the strains, and respectively obtaining 9 strains which are acid-resistant IAA-producing strains B3-54, O2-35 and N96; strains N134, N144 and N111 (figures 5-6) with phosphorus dissolving function; bacterial strains C3-11, N51 and O2-8 with potassium-decomposing function.
Example 3 test of the Protoffee effect of a Single functional bacterium on acid soil rape
Preparation of a microbial inoculum: 9 strains selected in example 2 above were used as test strains. Respectively inoculating 9 strains into TSB solid culture medium, culturing at 37 ℃ for 1d for activation, standing for later use, respectively placing the selected single colonies into LB liquid culture medium at 30 ℃ and shaking on a shaking table at 170rpm for 2d to obtain fermentation liquor, diluting and coating to count the number of thalli, wherein the concentration of the thalli of the fermentation liquor is more than or equal to 1 multiplied by 10 8 cfu/mL. Centrifuging the fermentation liquor at 4000rpm for 6min, removing the supernatant, and re-suspending the fermentation liquor by ten times the volume of sterile water to prepare the microbial inoculum.
Soaking rape seeds in warm water, airing for standby, and preparing seedling raising matrix, seedling raising tray and plastic basin. The Jiangxi soil (pH is 4.5) and quartz sand are mixed according to the volume ratio of 1:1, mixing to obtain mixed soil, wherein 300g of mixed soil is filled in each plastic basin for standby.
The test of this example was carried out in a greenhouse at 25℃and a relative humidity of 60% -70%, after emergence, the plant light supplement lamps of the greenhouse were turned on, at night 22:00 to the next morning 6: and (3) automatically turning off the 00 plant light supplementing lamp.
Dibbling rape seeds into seedling trays with seedling substrates, soaking 2-3 seeds in each hole, culturing for 4-5d, sprouting, spraying appropriate amount of chlormequat chloride (preventing formation of high seedlings), continuing culturing, and regularly watering (watering every 2d, the same shall apply below). After the rape seedlings grow to 3 true leaves, selecting the rape seedlings with more consistent growth vigor, transplanting the rape seedlings into a plastic pot (one rape seedling per pot) filled with mixed soil, watering the rape seedlings into the plastic pot after the transplanting is finished, and then watering the rape seedlings regularly.
Setting ten treatments of T1 (CK, clean water (sterile water)), T2 (B3-54), T3 (O2-35), T4 (N96), T5 (N134), T6 (N144), T7 (N111), T8 (C3-11), T9 (N51) and T10 (O2-8) for the rape seedlings transplanted into the plastic basin, setting 9 repetitions for each treatment, after the transplanted rape seedlings stably grow for one week, inoculating a microbial inoculum (the microbial inoculum is poured into mixed soil of the plastic basin and is close to the root of the rape seedlings), the inoculation proportion is 10% (100 mu L/g of mixed soil is 30mL microbial inoculum per plastic basin), continuing culturing after the bacterial watering is completed, growing 6 th true leaves after 3 weeks of culture, randomly selecting 3 rape seedlings for each treatment, measuring the fresh dry weight and chlorophyll content of the rape seedlings, and comparing the growth conditions of the seedlings of each treatment.
The measurement method of each index is as follows:
fresh weight measurement: cleaning the roots of rape seedlings with water, wiping with water-absorbing paper, and weighing;
Dry weight measurement: placing the weighed rape seedlings in an envelope, deactivating enzymes for 15min at 105 ℃, drying at 70 ℃ until the weight is constant, and weighing;
Chlorophyll content (SPAD) determination: and (5) determining chlorophyll content of the leaves at the upper, middle and lower parts of the rape seedling by using a SPAD tester, and taking an average value.
As can be seen from fig. 7 to 9, after 3 weeks of culturing of the watering bacteria, the growth indexes of the rape treated with T5 (N134), T6 (N144) and T7 (N111) were significantly better than those of the rape treated with T1 (CK). The fresh weight of T5 (N134) is 9.43g, and is improved by 3.76g compared with T1 (CK); next, T6 (N144), T4 (N96) and T7 (N111) were increased by 2.90g, 2.66g and 2.42g, respectively, as compared to T1 (CK). T7 (N111) has a dry weight of up to 0.56g, followed by T5 (N134) and T6 (N144), each of 0.54g. SPAD for T5 (N134) was largest, 39.15, followed by T6 (N144) and T7 (N111), SPAD 39.11 and 38.50, respectively. In conclusion, the fresh weight, the dry weight and the leaf SPAD value of the rape seedlings are improved to different degrees by T5 (N134), T6 (N144) and T7 (N111), and the rape seedling growth promoting agent has a good acid soil rape growth promoting effect.
Example 4 Protoffee test of combination of dominant single bacteria on acid soil rape
(1) The dominant functional growth promoting strains N134, N144, N111 screened in example 3 above were used as test strains (combined morphological features and 16S rRNA phylogenetic tree, N134, N144, N111 were all classified as Pseudomonas sp.).
(2) Dominant functional growth-promoting strain plate confrontation test
N134, N144 and N111 strains were inoculated into TSB solid medium, respectively, and cultured at 37℃for 1d for activation. The activated strains were streaked on TSB solid medium plates perpendicular to each other and placed in a 30℃incubator for 1d, and the growth and mutual inhibition of bacteria were observed, as shown in FIG. 10, in which the test strains did not antagonize each other.
(3) Preparation of microbial inoculum
Activating strains N134, N144 and N111 (the same as above) respectively, placing the selected single colonies in LB liquid medium at 30deg.C and shaking on a 170rpm shaking table for 2d respectively, obtaining N134 fermentation liquor, N144 fermentation liquor and N111 fermentation liquor, diluting and coating the number of bacterial cells, and the bacterial cell concentration of N134 fermentation liquor, N144 fermentation liquor and N111 fermentation liquor is not less than 1×10 8 cfu/mL.
Single microbial inoculum preparation: and (3) respectively centrifuging the N134 fermentation liquor, the N144 fermentation liquor and the N111 fermentation liquor at 4000rpm for 6min, removing the supernatant, and re-suspending the supernatant by using ten times the volume of sterile water of the fermentation liquor to prepare single microbial agents, namely an N134 microbial agent, an N144 microbial agent and an N111 microbial agent.
Preparing a composite microbial inoculum: mixing two or three of N134 microbial inoculum, N144 microbial inoculum and N111 microbial inoculum in equal volume to prepare the composite microbial inoculum.
(4) Test of seed rape in acid soil by using composite microbial inoculant
Soaking rape seeds in warm water, airing for standby, and preparing seedling raising matrix, seedling raising tray and plastic basin. The Jiangxi soil (pH is 4.5) and quartz sand are mixed according to the volume ratio of 1:1, mixing to obtain mixed soil, wherein 300g of mixed soil is filled in each plastic basin for standby.
The test of this example was carried out in a greenhouse at 25℃and a relative humidity of 60% -70%, after emergence, the plant light supplement lamps of the greenhouse were turned on, at night 22:00 to the next morning 6: and (3) automatically turning off the 00 plant light supplementing lamp.
Dibbling rape seeds into seedling trays with seedling substrates, soaking 2-3 seeds in each hole, culturing for 4-5d, sprouting, spraying appropriate amount of chlormequat chloride (preventing formation of high seedlings), continuing culturing, and regularly watering (watering every 2d, the same shall apply below). After the rape seedlings grow to 3 true leaves, selecting the rape seedlings with more consistent growth vigor, transplanting the rape seedlings into a plastic pot (one rape seedling per pot) filled with mixed soil, watering the rape seedlings into the plastic pot after the transplanting is finished, and then watering the rape seedlings regularly.
As shown in Table 1, the seedlings transplanted into plastic pots were subjected to 8 treatments, of which 3 single microbial agents, 4 composite microbial agents and 1 clear water (sterile water) Control (CK), each treatment was subjected to 9 replicates.
After the transplanted rape seedlings stably grow for one week, inoculating a microbial inoculum (the microbial inoculum is poured into mixed soil of a plastic basin and is close to the root of the rape seedlings), wherein the inoculation proportion is 10% (100 mu L/g dry weight of the mixed soil is 30mL of microbial inoculum is poured into each plastic basin), continuously culturing after finishing the fungus pouring, after culturing for 3 weeks, randomly selecting 3 rape seedlings after the 6 th true leaves grow, measuring fresh dry weight and chlorophyll content of the rape seedlings, and comparing the growth conditions of the treated rape seedlings. Each index was measured in the same manner as in example 3.
As can be seen from FIGS. 11 to 13, after 3 weeks of culturing of the watering bacteria, the fresh weight, dry weight and SPAD of the T5 (N134-N144) -treated seedlings were all the greatest. The fresh weight and the dry weight of the rape seedlings treated by the T5 (N134-N144) are obviously superior to those of the rape seedlings treated by the T1 (CK) and other single microbial agents and composite microbial agents. The T5 (N134-N144) treated rape seedlings were not significantly different from the T2 (N134) treated seedlings, but significantly better than the other treatments. In conclusion, the T5 (N134-N144) treatment has the optimal effect of promoting the growth of the acid soil rape.
Example 5 identification of strains N134 and N144
Strains N134, N144 were biologically identified using morphology and 16s rRNA gene sequence analysis.
As shown in FIG. 1, strain N134 was cultured on TSB solid medium plates at 30℃for 1-2d to form a yellowish brown, irregularly round, opaque, moist, relatively smooth-edged colony.
As shown in FIG. 2, strain N144 was cultured on TSB solid medium plates at 30℃for 1-2d to form yellowish brown, translucent, moist, and matte-edged colonies.
By comparing the 16s rRNA gene sequence of the strain N134 with similar sequences, a development tree is constructed. The results in FIG. 3 show that strain N134 has a higher homology with Pseudomonas and Pseudomonas bijieensis strain L-9 is in the same branch.
By comparing the 16s rRNA gene sequence of the strain N144 with similar sequences, a development tree is constructed. The results in FIG. 4 show that strain N144 has a higher homology with Pseudomonas and Pseudomonas migulae strain NBRC 103157 is in the same branch.
The strain N134 is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC NO. 29191.
The strain N144 is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC NO.29192.
Claims (9)
1. The Pseudomonas composite microbial agent is characterized by comprising Pseudomonas N134 and Pseudomonas N144, wherein the Pseudomonas N134 is classified and named as Pseudomonas sp N134, and is preserved in the China general microbiological culture Collection center (China general microbiological culture Collection center) with the preservation date of 2023 and 11 months and 30 days, and the preservation number of CGMCC NO.29191; the classification of the Pseudomonas N144 is named as Pseudomonas N144, and the Pseudomonas N144 is preserved in China general microbiological culture Collection center (CGMCC) with the preservation date of 2023, 11 and 30 and the preservation number of CGMCC NO.29192.
2. The pseudomonas composite microbial agent according to claim 1, wherein the composite microbial agent is prepared by the following steps:
(1) Respectively carrying out liquid fermentation on pseudomonas N134 with the preservation number of CGMCC No.29191 and pseudomonas N144 with the preservation number of CGMCC No.29192 to obtain pseudomonas N134 fermentation liquor and pseudomonas N144 fermentation liquor;
(2) Centrifuging the pseudomonas N134 fermentation liquor and the pseudomonas N144 fermentation liquor respectively, removing supernatant, and re-suspending with sterile water to obtain a pseudomonas N134 microbial agent and a pseudomonas N144 microbial agent;
(3) And mixing the pseudomonas N134 microbial inoculum and the pseudomonas N144 microbial inoculum in equal volumes to obtain the composite microbial inoculum.
3. The pseudomonas composite microbial agent according to claim 2, wherein the composite microbial agent is prepared by the following steps:
(1) Inoculating pseudomonas N134 with the preservation number of CGMCC NO.29191 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium comprises a TSB liquid culture medium, an LB liquid culture medium or an R2A liquid culture medium, and the liquid fermentation conditions are as follows: the temperature is 28-30 ℃, the rotating speed is 170-200rpm, the fermentation time is 1-2d, the pseudomonas N134 fermentation broth is obtained, the number of the coating and counting thalli is diluted, and the concentration of the thalli of the pseudomonas N134 fermentation broth is more than or equal to 1 multiplied by 10 8 cfu/mL;
Inoculating pseudomonas N144 with the preservation number of CGMCC NO.29192 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium comprises a TSB liquid culture medium, an LB liquid culture medium or an R2A liquid culture medium, and the liquid fermentation conditions are as follows: the temperature is 28-30 ℃, the rotating speed is 170-200rpm, the fermentation time is 1-2d, the pseudomonas N144 fermentation broth is obtained, the number of the coating and counting thalli is diluted, and the concentration of the thalli of the pseudomonas N144 fermentation broth is more than or equal to 1 multiplied by 10 8 cfu/mL;
(2) Centrifuging the pseudomonas N134 fermentation liquor, removing the supernatant, and re-suspending the supernatant by using sterile water with ten times the volume of the pseudomonas N134 fermentation liquor to obtain a pseudomonas N134 microbial inoculum;
centrifuging the pseudomonas N144 fermentation liquor, removing the supernatant, and re-suspending the supernatant by using sterile water with ten times the volume of the pseudomonas N144 fermentation liquor to obtain a pseudomonas N144 microbial inoculum;
(3) And mixing the pseudomonas N134 microbial inoculum and the pseudomonas N144 microbial inoculum in equal volumes to obtain the composite microbial inoculum.
4. The preparation method of the Pseudomonas composite microbial agent is characterized in that the composite microbial agent comprises Pseudomonas N134 and Pseudomonas N144, the classification of the Pseudomonas N134 is named as Pseudomonas sp N134, and the Pseudomonas N134 is preserved in China general microbiological culture Collection center (China general microbiological culture Collection center), the preservation date is 2023, 11 months and 30 days, and the preservation number is CGMCC NO.29191; the classification of the Pseudomonas N144 is named as Pseudomonas N144, and the Pseudomonas N144 is preserved in the China general microbiological culture Collection center (China Committee) for culture Collection of microorganisms, wherein the preservation date is 2023, 11 and 30, and the preservation number is CGMCC NO.29192;
The preparation method of the composite microbial inoculum comprises the following steps:
(1) Respectively carrying out liquid fermentation on pseudomonas N134 with the preservation number of CGMCC No.29191 and pseudomonas N144 with the preservation number of CGMCC No.29192 to obtain pseudomonas N134 fermentation liquor and pseudomonas N144 fermentation liquor;
(2) Centrifuging the pseudomonas N134 fermentation liquor and the pseudomonas N144 fermentation liquor respectively, removing supernatant, and re-suspending with sterile water to obtain a pseudomonas N134 microbial agent and a pseudomonas N144 microbial agent;
(3) And mixing the pseudomonas N134 microbial inoculum and the pseudomonas N144 microbial inoculum in equal volumes to obtain the composite microbial inoculum.
5. The preparation method of the pseudomonas composite microbial agent as claimed in claim 4, wherein the composite microbial agent is prepared by the following steps:
(1) Inoculating pseudomonas N134 with the preservation number of CGMCC NO.29191 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium comprises a TSB liquid culture medium, an LB liquid culture medium or an R2A liquid culture medium, and the liquid fermentation conditions are as follows: the temperature is 28-30 ℃, the rotating speed is 170-200rpm, the fermentation time is 1-2d, the pseudomonas N134 fermentation broth is obtained, the number of the coating and counting thalli is diluted, and the concentration of the thalli of the pseudomonas N134 fermentation broth is more than or equal to 1 multiplied by 10 8 cfu/mL;
Inoculating pseudomonas N144 with the preservation number of CGMCC NO.29192 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium comprises a TSB liquid culture medium, an LB liquid culture medium or an R2A liquid culture medium, and the liquid fermentation conditions are as follows: the temperature is 28-30 ℃, the rotating speed is 170-200rpm, the fermentation time is 1-2d, the pseudomonas N144 fermentation broth is obtained, the number of the coating and counting thalli is diluted, and the concentration of the thalli of the pseudomonas N144 fermentation broth is more than or equal to 1 multiplied by 10 8 cfu/mL;
(2) Centrifuging the pseudomonas N134 fermentation liquor, removing the supernatant, and re-suspending the supernatant by using sterile water with ten times the volume of the pseudomonas N134 fermentation liquor to obtain a pseudomonas N134 microbial inoculum;
centrifuging the pseudomonas N144 fermentation liquor, removing the supernatant, and re-suspending the supernatant by using sterile water with ten times the volume of the pseudomonas N144 fermentation liquor to obtain a pseudomonas N144 microbial inoculum;
(3) And mixing the pseudomonas N134 microbial inoculum and the pseudomonas N144 microbial inoculum in equal volumes to obtain the composite microbial inoculum.
6. Use of a pseudomonas composite bacterial agent according to any one of claims 1-3 for promoting the growth of sour soil canola.
7. The use according to claim 6, wherein the acidic soil pH is between 4.5 and 6.5.
8. The use according to claim 6, wherein the microbial inoculum is applied at a time after the rape seedlings are transplanted in an amount of 30-50 mL/plant rape seedlings.
9. The use according to claim 8, wherein the microbial inoculum is applied 7-10 days after the rape seedlings are transplanted, the application amount being 30-50 mL/plant rape seedlings.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410570976.6A CN118165892B (en) | 2024-05-09 | 2024-05-09 | Pseudomonas composite microbial agent and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410570976.6A CN118165892B (en) | 2024-05-09 | 2024-05-09 | Pseudomonas composite microbial agent and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN118165892A CN118165892A (en) | 2024-06-11 |
| CN118165892B true CN118165892B (en) | 2024-07-05 |
Family
ID=91352841
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202410570976.6A Active CN118165892B (en) | 2024-05-09 | 2024-05-09 | Pseudomonas composite microbial agent and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN118165892B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1111224A (en) * | 1994-05-06 | 1995-11-08 | 中国科学院新疆生物土壤沙漠研究所 | PGI bacteria suspension for activating rhizosphere soil phosphorus and its preparation |
| CN112175869A (en) * | 2020-09-29 | 2021-01-05 | 烟台市林业科学研究所 | Pseudomonas and application thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA3113021A1 (en) * | 2018-09-21 | 2020-03-26 | Pivot Bio, Inc. | Methods and compositions for improving phosphate solubilization |
| CN113817626B (en) * | 2020-12-07 | 2023-07-25 | 内蒙古百奥科技有限公司 | Pseudomonas brassicae Y1, culture method and application thereof, and soil conditioner |
| CN117264837B (en) * | 2023-10-09 | 2024-04-02 | 东北农业大学 | Pseudomonas with growth promoting function for low-temperature stress of plants and application thereof |
-
2024
- 2024-05-09 CN CN202410570976.6A patent/CN118165892B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1111224A (en) * | 1994-05-06 | 1995-11-08 | 中国科学院新疆生物土壤沙漠研究所 | PGI bacteria suspension for activating rhizosphere soil phosphorus and its preparation |
| CN112175869A (en) * | 2020-09-29 | 2021-01-05 | 烟台市林业科学研究所 | Pseudomonas and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN118165892A (en) | 2024-06-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105385638B (en) | A kind of microorganism phosphorus decomposing preparation and its preparation method and application | |
| CN104726378B (en) | The method for improving salt stress turfgrass defence enzyme activity using Salt-tolerant microbial agent is strengthened | |
| CN104818233A (en) | Bacillus vallismortis and functional vegetable seedling raising biological matrix prepared from bacillus vallismortis | |
| CN113943660A (en) | Talaromyces fungus NJAU-L8 for preventing and controlling continuous cropping soil-borne blight and application thereof | |
| CN111690578A (en) | Salt and alkali resistant Siamese bacillus and production method and application of viable bacteria preparation thereof | |
| CN101519644B (en) | Sinorhizobium sp. and application thereof | |
| CN113322209A (en) | Plant growth-promoting composite microbial inoculum produced based on ecological relationship among strains and application thereof | |
| CN116463220B (en) | Dark-color DSE fungus for promoting blueberry growth and application thereof | |
| CN110205249B (en) | Method for promoting plant growth and alternaria alternate fungus used by same | |
| CN115725419B (en) | Phosphorus-dissolving blueberry endophytic trichoderma and application thereof | |
| CN118165892B (en) | Pseudomonas composite microbial agent and application thereof | |
| CN118147017B (en) | Composite microbial inoculum containing bacteria and fungi and application thereof | |
| CN118147016B (en) | Pseudomonas N134 and application thereof | |
| CN110241040B (en) | Korean pseudomonas and application thereof in improving organic nitrogen utilization rate of facility vegetable soil and promoting growth | |
| CN113897316A (en) | Bacillus licheniformis BLc06, functional melon and fruit seedling biological matrix prepared from same and application of functional melon and fruit seedling biological matrix | |
| CN117802012B (en) | Ammonia philic bacteria, bacterial agent and application thereof | |
| CN104798819B (en) | A kind of method for improving turfgrass anti-seismic design using low temperature resistant microbial bacterial agent is strengthened | |
| CN108531428A (en) | A kind of preparation method of fast-growing frankia bacterium powder | |
| CN116836896B (en) | Microbial agent for preventing and treating cotton wilt and preparation method thereof | |
| CN117586931B (en) | Achromobacter xylosoxidans IVF-WK240 and application thereof | |
| CN115851524B (en) | Strain for increasing crop yield and application thereof | |
| CN113215000B (en) | Chaetomium, seed soaking agent and application | |
| CN113249266B (en) | Bacillus subtilis capable of efficiently dissolving phosphorus and promoting sugarcane germination and application thereof | |
| Situmorang | ISOLATION AND NODULATION TEST OF Rhizobium sp. FROM Pueraria javanica (Benth.) Benth. AND LIABILITY TEST ON THE CARRIER MEDIUM OF PEAT AND COMPOST FROM PALM OIL PALM EMPTY FRUITS | |
| Siahbalaei et al. | New morphospecies of Phormidium and Microcoleus (Cyanophyta) in paddy-fields of Iran |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |