CN114276950B - Wide-pH-adaptability salt-tolerant phosphate solubilizing bacterium and application thereof - Google Patents
Wide-pH-adaptability salt-tolerant phosphate solubilizing bacterium and application thereof Download PDFInfo
- Publication number
- CN114276950B CN114276950B CN202111415216.0A CN202111415216A CN114276950B CN 114276950 B CN114276950 B CN 114276950B CN 202111415216 A CN202111415216 A CN 202111415216A CN 114276950 B CN114276950 B CN 114276950B
- Authority
- CN
- China
- Prior art keywords
- pseudomonas
- phosphate
- suaeda salsa
- solubilizing
- seeds
- 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
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Pretreatment Of Seeds And Plants (AREA)
Abstract
The invention discloses a wide-range pH salt-tolerant phosphate-solubilizing bacterium and application thereof, wherein the wide-range pH salt-tolerant phosphate-solubilizing bacterium is pseudomonas PSBDYJP20210001, can tolerate the environmental conditions of pH 2-10 and NaCl concentration 1-5%, and has a higher phosphate-solubilizing function, and the pseudomonas PSBDYJP20210001 is preserved in: china center for type culture Collection, the collection addresses are: wuhan, china, the preservation date is: 13/09/2021, accession number: CCTCC NO: M20211169. The seeds of the suaeda salsa are soaked in the culture solution of the pseudomonas PSBDYJP20210001, so that the germination rate of the seeds can be obviously improved; the culture solution of the pseudomonas PSBDYJP20210001 is used for irrigating the soil for planting the suaeda salsa, so that the plant height, the root length, the root diameter and the root tip number of the suaeda salsa can be obviously improved.
Description
Technical Field
The invention relates to a strain and application thereof, in particular to a salt-tolerant phosphate-solubilizing bacterium with wide pH value and application thereof, belonging to the technical field of microorganisms.
Background
Phosphorus is one of mineral elements necessary for plant growth and development, is an important material basis for plant growth, and the amount of phosphorus absorbed by plants is in positive correlation with biomass and yield thereof. Most of phosphorus in the soil exists in an inorganic phosphorus or organic phosphide state and cannot be absorbed and utilized by plants. Phosphate Solubilizing bacteria (Phosphate Solubilizing bacteria) are bacteria which can convert insoluble phosphorus into phosphorus which can be absorbed and utilized by plants so as to improve the phosphorus absorption of the plants and further promote the plant growth, and the screening of the Phosphate Solubilizing bacteria with the characteristic of high-efficiency Phosphate solubilization has become a hot research topic in the technical field of agricultural microorganisms in China.
The total area of the saline-alkali soil in China exceeds about 14.87 hundred million mu, and accounts for 4.88 percent of the total available soil area in China. Meanwhile, the extensive expansion of facility agriculture in coastal areas leads to excessive exploitation of underground water, further to the salinity exceeding of seawater, cultivated soil and irrigation water, and seriously restricts the growth of terrestrial vegetation. The phosphate solubilizing bacteria are utilized to dissolve and release phosphorus in soil, promote plant growth and have positive ecological significance for improving land utilization efficiency.
At present, the reported phosphate solubilizing bacteria, whether belonging to different genera or different strains belonging to the same genus, have different phosphate solubilizing efficiencies, different phosphate solubilizing mechanisms and different application effects in saline-alkali soil. Therefore, screening the phosphate solubilizing bacteria which can tolerate acid and alkali and high-salt environments and exploring the optimal application amount of the phosphate solubilizing bacteria have important ecological significance for promoting plant growth and improving land utilization efficiency.
Disclosure of Invention
The invention aims to provide a phosphate solubilizing bacterium with wide pH adaptation and salt tolerance, which can resist acid and alkali and high-salt environment.
The second purpose of the invention is to provide the application of the phosphate-solubilizing bacteria with wide pH adaptation in the environment with the pH of 2-10 and the mass concentration of NaCl of 1-5%.
In order to achieve the first object, the invention adopts the following technical scheme:
the wide-adaptation pH salt-tolerant phosphate-solubilizing bacterium is Pseudomonas PSBDYJP20210001 with Latin name of Pseudomonas sp.PSBDYJP20210001, can tolerate the environmental conditions of pH 2-10 and NaCl concentration 1-5%, and has a higher phosphate-solubilizing function, and the Pseudomonas PSBDYJP20210001 is preserved in: china center for type culture Collection, the collection addresses are: wuhan, china, the preservation date is: 13/09/2021, accession number: CCTCC NO: M20211169.
In order to achieve the second objective, the invention adopts the following technical scheme:
1. the application of the phosphate-tolerant phosphate-solubilizing bacterium with the broad pH value (Pseudomonas PSBDYJP 20210001) in promoting the germination of the suaeda salsa seeds is characterized in that the step of promoting the germination of the suaeda salsa seeds comprises the following steps:
(1) Culturing pseudomonas PSBDYJP20210001 to logarithmic phase by using LB enrichment medium, then inoculating the pseudomonas PSBDYJP 5363 to NBRIP liquid culture medium, and then performing shake culture at 30 ℃ and 220r/min for 24h to obtain phosphate solubilizing bacteria liquid;
(2) Sterilizing Suaeda salsa seeds;
(3) And (3) soaking the sterilized suaeda salsa seeds obtained in the step (2) by the phosphate solubilizing bacteria liquid obtained in the step (1) for 5 days.
2. The application of the phosphate-solubilizing bacteria with wide pH adaptation (Pseudomonas PSBDYJP 20210001) in promoting the growth of the Suaeda salsa plants is characterized in that the step of promoting the growth of the Suaeda salsa plants comprises the following steps:
(1) Culturing pseudomonas PSBDYJP20210001 to logarithmic phase by using LB enrichment medium, then inoculating the pseudomonas PSBDYJP 5363 to NBRIP liquid culture medium, and then performing shake culture at 30 ℃ and 220r/min for 24h to obtain phosphate solubilizing bacteria liquid;
(2) Sterilizing Suaeda salsa seeds, and then putting the Suaeda salsa seeds into an incubator at 28 ℃ for culturing until the seeds germinate;
(3) Irrigating the soil with water to enable the water content of the soil to reach 80% of the maximum water holding capacity, then irrigating the soil with the phosphate solubilizing bacterium liquid obtained in the step (1), and then planting the germinated suaeda salsa seeds obtained in the step (2) into the soil within 24 hours.
The invention has the advantages that:
(1) The pseudomonas PSBDYJP20210001 which is the salt-tolerant phosphate-solubilizing bacterium with the wide pH range and is obtained by screening can tolerate the environmental conditions of pH 2-10 and NaCl concentration 1-5% and has a higher phosphate-solubilizing function; the culture solution of the pseudomonas PSBDYJP20210001 is used for soaking the seeds of the suaeda salsa, so that the germination rate of the seeds of the suaeda salsa can be obviously improved; the culture solution of the pseudomonas PSBDYJP20210001 is used for irrigating the soil for planting the suaeda salsa, so that the plant height, the root length, the root diameter and the root tip number of the suaeda salsa can be obviously improved.
(2) The pseudomonas PSBDYJP20210001 which is the salt-tolerant phosphate-solubilizing bacteria with the wide-adaptation pH value and is obtained by screening promotes the absorption and utilization of plants by converting insoluble or insoluble phosphorus in soil into quick-acting phosphorus or soluble phosphorus to be stored in the soil, thereby promoting the growth of the plants and having positive ecological significance for improving the land utilization efficiency.
Drawings
FIG. 1 is a phosphorus-solubilizing circle diagram of Pseudomonas PSBDYJP20210001 screened by the invention;
FIG. 2 is a phylogenetic tree of Pseudomonas PSBDYJP20210001 screened by the present invention;
FIG. 3 is a graph showing the phosphate solubilizing capacity of Pseudomonas PSBDYJP20210001 screened by the present invention in the range of pH =2 to 10;
FIG. 4 is a graph showing the phosphorus-solubilizing capacity of Pseudomonas PSBDYJP20210001 screened by the present invention in the range of NaCl mass concentration of 1 to 5%.
Detailed Description
The invention is described in detail below with reference to the figures and the embodiments.
1. Culture medium
1. Meng Jinna inorganic phosphorus solid culture medium
5363 formulation of inorganic phosphorus solid culture medium Meng Jinna is as follows: glucose 10g, (NH) 4 ) 2 SO 4 0.5g，NaCl 10g，KCl 0.3g，MgSO 4 ·7H 2 O 0.3g，FeSO 4 ·7H 2 0 0.036g，MnS0 4 ·4H 2 O 0.03g，Ca 3 (P0 4 ) 2 5g, agar 18g, distilled water 1000mL, tris-HCl adjusted pH 8.
2. LB enrichment medium
The formulation of LB enriched medium is as follows: 10g of tryptone, 5g of yeast powder, 10g of NaCl and 1000mL of distilled water.
3. NBRIP liquid medium
The formula of the NBRIP liquid culture medium is as follows: glucose was 10g, ca 3 (P0 4 ) 2 5g，MgCl 2 5g，MgSO 4 0.25g，KCl 0.2g，(NH 4 ) 2 SO 4 0.1g, 1000mL of distilled water, and the pH was adjusted to 7.0.
2. Separation, purification and identification of wide-pH-range salt-tolerant phosphate-solubilizing bacteria-pseudomonas PSBDYJP20210001
1. Separation and purification of pseudomonas PSBDYJP20210001
The soil sample is taken from saline-alkali soil of east-Shandong yellow river mouth, 10g of the soil sample is placed in a 500mL conical flask filled with 90mL of distilled water, ten-fold gradient dilution is carried out on the mixed solution, proper dilution is taken and respectively coated on Meng Jinna inorganic phosphorus solid culture media (flat plates), the mixed solution is cultured for 4 days at 30 ℃ (figure 1), a bacterial colony with an obvious phosphorus dissolving ring is selected and subjected to streak purification, the bacterial colony is numbered PSBDYJP20210001, and the bacterial colony is stored for later use at 4 ℃.
The strain PSBDYJP20210001 is shake-cultured in LB enrichment medium at 30 ℃ and 220r/min, and the strain is preserved when the bacterial liquid is turbid and the bacterial liquid is hung on the wall.
2. Identification of Strain PSBDYJP20210001
16S rDNA amplification, sequencing and splicing are carried out on a strain PSBDYJP20210001 to obtain a gene sequence, BLAST analysis is carried out on the gene sequence and a microbial sequence in a Genbank database to find that the homology of the 16S rDNA sequence of the strain and pseudomonas is more than 99 percent, the sequence of the strain is uploaded to the Genbank database, and the accession number is OL305679.
Phylogenetic trees were constructed using the MEGA adjacency method, and the results are shown in FIG. 2. FIG. 2 shows that: the strain PSBDYJP20210001 is integrated with pseudomonas into one.
3. Determination of phosphate solubilizing Activity of Pseudomonas PSBDYJP20210001
Activating pseudomonas PSBDYJP20210001 by adopting LB enrichment medium, culturing to logarithmic phase at 30 ℃, then inoculating into a conical flask filled with 100mL NBRIP liquid culture medium according to the inoculum size of 10%, and finally performing shake culture for 24h at 30 ℃ by a shaking table at 220 r/min.
Values were recorded as 0mg, 0.2mg, 0.4mg, 0.6mg, 0.8mg and 1.0mg phosphorus standard series solutions simultaneously with the blank at a wavelength of 700 nm. And drawing a phosphorus standard curve by taking the measured absorbance as a vertical coordinate and the phosphorus concentration as a vertical coordinate.
In order to measure the phosphorus dissolving capacity of the pseudomonas PSBDYJP20210001, the phosphorus dissolving amount of the pseudomonas PSBDYJP20210001 is measured by adopting a molybdenum-antimony-scandium colorimetric method, calcium phosphate is the only inorganic phosphorus source, and the inorganic phosphorus dissolving capacity of the pseudomonas PSBDYJP20210001 is measured.
The phosphorus solubilizing ability of pseudomonas PSBDYJP20210001 and other phosphorus solubilizing bacteria is compared in Table 1.
TABLE 1 comparison of phosphate solubilizing Capacity
| Bacterial strains | PSBDYJP20210001 | QDZN-13 | DC-3-P |
| Phosphorus amount (mg/L) | 182±7.5 | 175.83±1.2 | 158.24 |
As can be seen from Table 1: the phosphate solubilizing performance of the pseudomonas PSBDYJP20210001 is stronger than that of common phosphate solubilizing bacteria.
4. Determination of acid and alkali resistance range of pseudomonas PSBDYJP20210001
In order to measure the acid and alkali resistance range of the pseudomonas PSBDYJP20210001, the pseudomonas PSBDYJP20210001 is cultured to the logarithmic growth phase by using an LB enrichment medium, then the pseudomonas PSBDYJP20210001 is inoculated into a conical flask filled with 100mL NBRIP liquid culture medium in a 10% inoculation amount, the pH value is adjusted by using HCl and NaOH, then the liquid culture medium is shake-cultured for 24h at 30 ℃ and 220r/min, finally the phosphorus removal amount of the pseudomonas PSBDYJP20210001 is measured by adopting a molybdenum-antimony-scandium colorimetric method, calcium phosphate is a unique phosphorus source of inorganic phosphorus, and the inorganic phosphorus removal capacity of the pseudomonas PSBDYJP20210001 is measured.
The phosphorus-solubilizing amount of pseudomonas PSBDYJP20210001 in the range of pH = 2-10 is shown in fig. 3.
The acid and alkali resistance range (pH) of the pseudomonas PSBDYJP20210001 and other phosphate solubilizing bacteria is compared in Table 2.
TABLE 2 acid and alkali resistance Range (pH) comparison
| Bacterial strains | PSBDYJP20210001 | QL1501 |
| Acid and alkali resistance range (pH) | 2～10 | 7～9 |
The phosphorus-dissolving amount of the pseudomonas PSBDYJP20210001 and other phosphorus-dissolving bacteria is compared in table 3.
TABLE 3 comparison of phosphorus solubilizing amounts
As can be seen from fig. 3 and tables 2 and 3: the acid-base resistance range and the phosphate-solubilizing quantity of the pseudomonas PSBDYJP20210001 are both stronger than those of common phosphate-solubilizing bacteria.
5. Influence of pseudomonas PSBDYJP20210001 culture solution (phosphate solubilizing bacterium solution) with different pH values on germination rate of suaeda salsa seeds
Culturing pseudomonas PSBDYJP20210001 to a logarithmic phase by using an LB enrichment medium, then adjusting the pH value to be 2, 3, 4, 5, 6, 7, 8, 9 and 10 by adding HCl and NaOH, and then performing shake culture at 30 ℃ and 220r/min for 24h to obtain pseudomonas PSBDYJP20210001 culture solutions (namely phosphate solubilizing bacteria solutions with different pH values) with different pH values for later use.
Seed disinfection: sterilizing Suaeda salsa seeds with 70% alcohol for 1min, sterilizing Suaeda salsa seeds with 1% sodium hypochlorite for 15min, and washing the Suaeda salsa seeds with sterile water for 5 times.
Taking 30 sterilized culture dishes, dividing the culture dishes into 10 groups, wherein each group comprises 3 culture dishes, and 50 sterilized suaeda salsa seeds are placed in each culture dish. Adding the obtained 9 parts of phosphate-solubilizing bacteria liquid with different pH values (the pH values are respectively 2, 3, 4, 5, 6, 7, 8, 9 and 10) into the first 9 groups of culture dishes respectively to ensure the suaeda salsa seeds without salt, adding the Meng Jinna inorganic phosphorus liquid culture medium into the 10 th group of culture dishes (used as a control) to ensure the suaeda salsa seeds without salt, placing all the culture dishes into a 28 ℃ culture box, and waiting for the suaeda salsa seeds to germinate. The germination rate of the soaked suaeda salsa seeds in each culture dish is counted by regular observation every day.
The experiment was repeated 3 times and the results averaged.
The influence of the phosphate solubilizing bacteria solutions with different pH values on the germination rate of suaeda salsa is shown in Table 4.
TABLE 4 influence of phosphate solubilizing bacteria solutions with different pH values on germination rate of Suaeda salsa seeds
Note: the lower case letters in the same column indicate significant differences (P < 0.05).
As can be seen from table 4: when the pH value of the phosphate-solubilizing bacterial liquid is 5, 6 or 7, the germination rate of the suaeda salsa seeds is obviously higher than that of a control group; when the pH value of the phosphate-solubilizing bacterial liquid is 5, the germination rate of the suaeda salsa seeds is obviously higher than that of the other phosphate-solubilizing bacterial liquids with different pH values (p is less than 0.05).
6. Influence of pseudomonas PSBDYJP20210001 culture solution (phosphate solubilizing bacterium solution) with different pH values on the character of suaeda salsa plants
Test soil: is taken from saline-alkali soil of east Shandong Ying yellow river estuary.
Phosphate solubilizing bacteria: pseudomonas PSBDYJP20210001.
Plant: suaeda salsa.
The soil to be tested is filled into plastic pots with the height of 11cm, the pot mouth diameter of 13cm and the pot bottom diameter of 9cm, the dry weight of each pot of soil is 1kg, and distilled water is used for supplementing the soil moisture, so that the soil moisture content reaches 80% of the maximum water holding capacity.
Culturing the pseudomonas PSBDYJP20210001 to a logarithmic phase by using an LB enrichment medium, then adjusting the pH value by using HCl and NaOH, wherein the pH value is respectively 2, 3, 4, 5, 6, 7, 8, 9 and 10, and then performing shake culture at 30 ℃ and 220r/min for 24h to obtain pseudomonas PSBDYJP20210001 culture solutions (i.e. phosphate solubilizing bacteria solutions with different pH values), and meanwhile, setting the LB enrichment medium without adding the pseudomonas PSBDYJP20210001 as a control group.
Seed disinfection: sterilizing Suaeda salsa seeds with 70% alcohol for 1min, sterilizing Suaeda salsa seeds with 1% sodium hypochlorite for 15min, and washing the Suaeda salsa seeds with sterile water for 5 times.
Taking the sterilized culture dish, putting the sterilized suaeda salsa seeds into the culture dish, and putting the culture dish in an incubator at 28 ℃ for culturing until the seeds germinate.
Respectively irrigating an LB enrichment medium and phosphate solubilizing bacteria liquid with different pH values into different plastic pots, wherein each pot is 100mL, planting the germinated suaeda salsa seeds into the plastic pots within 24h, sowing 50 seeds in each pot, and measuring and counting the plant height of plants after sowing for 20 days; and meanwhile, a root system scanner is adopted to analyze the root system of the plant, and the root length, the root system diameter and the root tip number are specifically related.
The experiment was repeated 3 times and the results averaged.
The effect of the phosphate solubilizing bacteria solutions with different pH values on the plant characteristics of suaeda salsa is shown in Table 5.
TABLE 5 influence of the phosphate solubilizing bacteria solutions with different pH values on overground and underground growth of Suaeda salsa
Note: the lower case letters in the same column indicate significant difference (P < 0.05).
As can be seen from table 5: when the pH of the phosphate-solubilizing bacterial liquid is 4, 5, 6 and 7, the plant height, the root length, the root diameter and the root tip number of the suaeda salsa plant are all higher than those of the control group; when the pH of the phosphate-solubilizing bacterial liquid is 2-9, the plant height and the root length of the suaeda salsa plant are both higher than those of the control group; when the pH of the phosphate-solubilizing bacterial liquid is 5, the plant height, the root length, the root diameter and the root tip number of the suaeda salsa plants are all obviously higher than those of the other phosphate-solubilizing bacterial liquids with different pH values (p is less than 0.05).
7. Determination of salinity tolerance Range of Pseudomonas PSBDYJP20210001
In order to measure the salinity tolerance range of the pseudomonas PSBDYJP20210001d, the salt concentration range is adjusted by adding NaCl, the phosphorus dissolving amount of the pseudomonas PSBDYJP20210001 is measured by adopting a molybdenum-antimony-scandium colorimetric method, calcium phosphate is the only phosphorus source of inorganic phosphorus, and the inorganic phosphorus dissolving capacity of the pseudomonas PSBDYJP20210001 is measured.
The phosphorus decomposition amount of the pseudomonas PSBDYJP20210001 under different salt concentration conditions is shown in figure 4.
The phosphorus-dissolving amount of the pseudomonas PSBDYJP20210001 and other phosphorus-dissolving bacteria under different salt concentration conditions is compared in a table 6.
TABLE 6 comparison of phosphorus solubilizing amounts
As can be seen from fig. 4 and table 6: the salinity tolerance range of the pseudomonas PSBDYJP20210001 is stronger than that of common phosphate solubilizing bacteria.
8. Influence of pseudomonas PSBDYJP20210001 culture solution (phosphate solubilizing bacterium solution) with different salt concentrations on germination rate of suaeda salsa seeds
Culturing pseudomonas PSBDYJP20210001 to a logarithmic phase by using an LB enrichment medium, then inoculating 10% of inoculum size into a triangular flask filled with 100mL of NBRIP liquid culture medium, adjusting the salt concentration range by adding NaCl, wherein the salt concentration is 1%, 2%, 3%, 4% and 5%, and then performing shake culture at 30 ℃ and 220r/min for 24h to obtain pseudomonas PSBDYJP20210001 culture solutions (namely phosphate solubilizing bacteria solutions with different salt concentrations) with different salt concentrations for later use.
Seed disinfection: sterilizing Suaeda salsa seeds with 70% alcohol for 1min, sterilizing Suaeda salsa seeds with 1% sodium hypochlorite for 15min, and washing Suaeda salsa seeds with sterile water for 5 times.
Taking 30 sterilized culture dishes, dividing the culture dishes into 6 groups, wherein each group comprises 5 culture dishes, and 50 sterilized suaeda salsa seeds are put into each culture dish. Respectively adding the phosphate-solubilizing bacteria liquid with the 5 salt concentrations (the salt concentrations are 1%, 2%, 3%, 4% and 5%) obtained in the above step into the first 5 groups of culture dishes to ensure that suaeda glauca seeds have no salt, adding Meng Jinna inorganic phosphorus liquid culture medium into the 6 th group of culture dishes (serving as a control) to ensure that the suaeda glauca seeds have no salt, placing all the culture dishes into a 28 ℃ culture box, and waiting for the seeds to germinate. The germination rate of the soaked suaeda salsa seeds in each culture dish is counted by regular observation every day.
The experiment was repeated 3 times and the results averaged.
The influence of the phosphate solubilizing bacteria solutions with different salt concentrations on the germination rate of the suaeda salsa seeds is shown in table 7.
TABLE 7 influence of phosphate solubilizing bacteria solutions with different salt concentrations on germination rate of Suaeda salsa seeds
Note: the lower case letters in the same column indicate significant difference (P < 0.05).
As can be seen from Table 7: on the 2 nd day of seed soaking, the phosphate solubilizing bacteria liquid with 1% salt concentration already shows better germination accelerating effect than the phosphate solubilizing bacteria liquid with other salt concentrations; from the 3 rd day of soaking the seeds, the effect of accelerating germination of the phosphate solubilizing bacterial liquid with the salt concentration of 2% on the suaeda salsa seeds is better than that of a control group; on the 5 th day of seed soaking, the phosphate-solubilizing bacteria liquid of the soaked seeds still keeps high germination accelerating effect (p is less than 0.05).
9. Influence of pseudomonas PSBDYJP20210001 phosphate solubilizing bacterial liquid with different salt concentrations on saline land suaeda salsa plant characters
Test soil: is prepared from saline-alkali soil of Dongying yellow river estuary.
Phosphate solubilizing bacteria: pseudomonas PSBDYJP20210001.
Plant: suaeda salsa.
The soil to be tested is filled into plastic pots with the height of 11cm, the pot mouth diameter of 13cm and the pot bottom diameter of 9cm, the dry weight of each pot of soil is 1kg, and distilled water is used for supplementing the soil moisture, so that the soil moisture content reaches 80% of the maximum water holding capacity.
Culturing the pseudomonas PSBDYJP20210001 to a logarithmic phase by using an LB enrichment culture medium, then adjusting the salt concentration range by using NaCl, wherein the salt concentration is respectively 1%, 2%, 3%, 4% and 5%, and then performing shake culture at 30 ℃ and 220r/min for 24h to obtain pseudomonas BDPSYJP 20210001 culture solutions (namely, phosphate solubilizing bacteria solutions with different salt concentrations) with different salt concentrations, and meanwhile, setting the LB enrichment culture medium without adding the pseudomonas PSBDYJP20210001 as a control group.
Seed disinfection: sterilizing Suaeda salsa seeds with 70% alcohol for 1min, sterilizing Suaeda salsa seeds with 1% sodium hypochlorite for 15min, and washing the Suaeda salsa seeds with sterile water for 5 times.
Taking the sterilized culture dish, putting the sterilized suaeda salsa seeds into the culture dish, and putting the culture dish in an incubator at 28 ℃ for culturing until the seeds germinate.
Respectively irrigating an LB enrichment culture medium and phosphate solubilizing bacteria liquid with different salt concentrations into different plastic pots, wherein each pot is 100mL, planting the germinated suaeda salsa seeds into the plastic pots within 24h, sowing 50 seeds in each pot, and measuring and counting the plant height of plants after sowing for 20 days; and meanwhile, a root system scanner is adopted to analyze the root system of the plant, and the root length, the root system diameter and the root tip number are specifically related.
The experiment was repeated 3 times and the results averaged.
The effect of the phosphate solubilizing bacteria liquid with different salt concentrations on the plant characteristics of suaeda salsa is shown in table 8.
TABLE 8 influence of phosphate solubilizing bacteria solutions of different salt concentrations on the plant characteristics of Suaeda salsa
Note: the lower case letters in the same column indicate significant differences (P < 0.05).
As can be seen from Table 8, when the salt concentration of the phosphate-solubilizing bacterial liquid is 1%, 2% or 3%, the plant height, root length, root diameter and root tip number of the Suaeda salsa plants are all higher than those of the control group; when the salt concentration of the phosphate-solubilizing bacterial liquid is 1%, the plant height, the root length, the root diameter and the root tip number of the suaeda salsa plants are all obviously higher than those of the phosphate-solubilizing bacterial liquid with the rest salt concentration (p is less than 0.05).
In conclusion, the Pseudomonas PSBDYJP20210001, which is the broad-adaptation pH salt-tolerant phosphate solubilizing bacterium screened by the invention, can tolerate the environmental conditions of pH 2-10 and NaCl concentration 1-5%, and has a higher phosphate solubilizing function; the culture solution of the pseudomonas PSBDYJP20210001 is used for soaking the seeds of the suaeda salsa, so that the germination rate of the seeds of the suaeda salsa can be obviously improved; the culture solution of the pseudomonas PSBDYJP20210001 is used for irrigating the soil for planting the suaeda salsa, so that the plant height, the root length, the root diameter and the root tip number of the suaeda salsa can be obviously improved.
The pseudomonas PSBDYJP20210001 which is the salt-tolerant phosphate-solubilizing bacteria with the wide-adaptation pH value and is obtained by screening promotes the absorption and utilization of plants by converting insoluble or insoluble phosphorus in soil into quick-acting phosphorus or soluble phosphorus to be stored in the soil, thereby promoting the growth of the plants and having positive ecological significance for improving the land utilization efficiency.
It should be noted that the above-mentioned embodiments do not limit the present invention in any way, and all technical solutions obtained by using equivalent alternatives or equivalent variations fall within the protection scope of the present invention.
Claims (4)
1. A strain of wide-adaptation pH salt-tolerant phosphate-solubilizing bacteria is characterized in that the wide-adaptation pH salt-tolerant phosphate-solubilizing bacteria is pseudomonas PSBDYJP20210001 with the Latin namePseudomonassp, PSBDYJP20210001, able to withstand environmental conditions of pH 2-10 and NaCl concentration 1-5%, while having a high phosphate solubilizing function, the pseudomonad PSBDYJP20210001 deposited: china center for type culture Collection, the collection addresses are: wuhan, china, the preservation date is: 13/09/2021, accession number: CCTCC NO: M20211169.
2. The application of the wide-pH-range salt-tolerant phosphate solubilizing bacteria as claimed in claim 1, wherein the step of promoting the germination of the suaeda salsa seeds comprises the following steps:
(1) Culturing pseudomonas PSBDYJP20210001 to logarithmic phase by using LB enrichment medium, then inoculating the pseudomonas PSBDYJP 5363 to NBRIP liquid culture medium, and then performing shake culture at 30 ℃ and 220r/min for 24h to obtain phosphate solubilizing bacteria liquid;
(2) Sterilizing Suaeda salsa seeds;
(3) And (3) soaking the disinfected suaeda salsa seeds obtained in the step (2) in the phosphate-solubilizing bacterial liquid obtained in the step (1) for 5 days.
3. The use of the salt-tolerant phosphate-solubilizing bacteria with a broad pH range as claimed in claim 1 for promoting the growth of Suaeda salsa plants, wherein the step of promoting the growth of Suaeda salsa plants comprises:
(1) Culturing pseudomonas PSBDYJP20210001 to logarithmic phase by using LB enrichment medium, then inoculating the pseudomonas PSBDYJP 5363 to NBRIP liquid culture medium, and then performing shake culture at 30 ℃ and 220r/min for 24h to obtain phosphate solubilizing bacteria liquid;
(2) Sterilizing Suaeda salsa seeds, and then putting the Suaeda salsa seeds into an incubator at 28 ℃ for culturing until the seeds germinate;
(3) Irrigating the soil with water to enable the water content of the soil to reach 80% of the maximum water holding capacity, then irrigating the soil with the phosphate solubilizing bacterium liquid obtained in the step (1), and then planting the germinated suaeda salsa seeds obtained in the step (2) into the soil within 24 hours.
4. The use of claim 3, wherein in step (3), the amount of the phosphate-solubilizing bacterial liquid poured is 100mL/kg.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111415216.0A CN114276950B (en) | 2021-11-25 | 2021-11-25 | Wide-pH-adaptability salt-tolerant phosphate solubilizing bacterium and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111415216.0A CN114276950B (en) | 2021-11-25 | 2021-11-25 | Wide-pH-adaptability salt-tolerant phosphate solubilizing bacterium and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114276950A CN114276950A (en) | 2022-04-05 |
| CN114276950B true CN114276950B (en) | 2023-01-10 |
Family
ID=80869857
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111415216.0A Active CN114276950B (en) | 2021-11-25 | 2021-11-25 | Wide-pH-adaptability salt-tolerant phosphate solubilizing bacterium and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114276950B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116333924B (en) * | 2023-02-22 | 2024-03-22 | 鲁东大学 | Biological organic conditioner special for saline-alkali soil and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108496616A (en) * | 2018-02-12 | 2018-09-07 | 暨南大学 | A method of promote agrobacterium to be colonized in plant rhizosphere |
| CN109679858A (en) * | 2018-09-07 | 2019-04-26 | 山东省科学院生态研究所 | The pseudomonas fluorescens strain and its cultural method of one plant of phosphorus decomposing salt tolerant and application |
| CN113373096A (en) * | 2021-07-21 | 2021-09-10 | 盐城师范学院 | Bacillus cereus and application thereof in relieving plant salt stress |
-
2021
- 2021-11-25 CN CN202111415216.0A patent/CN114276950B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108496616A (en) * | 2018-02-12 | 2018-09-07 | 暨南大学 | A method of promote agrobacterium to be colonized in plant rhizosphere |
| CN109679858A (en) * | 2018-09-07 | 2019-04-26 | 山东省科学院生态研究所 | The pseudomonas fluorescens strain and its cultural method of one plant of phosphorus decomposing salt tolerant and application |
| CN113373096A (en) * | 2021-07-21 | 2021-09-10 | 盐城师范学院 | Bacillus cereus and application thereof in relieving plant salt stress |
Non-Patent Citations (2)
| Title |
|---|
| Unraveling bacterial community structure and function and their links with natural salinity gradient in the Yellow River Delta;Zifang Chi;《Sci Total Environ》;20210208;第773卷;第1-9页 * |
| 具ACC脱氨酶活性的碱蓬内生细菌的分离、鉴定及其生物学特性;滕松山等;《微生物学报》;20101231;第50卷(第11期);第1503-1509页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114276950A (en) | 2022-04-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103275891B (en) | Endophyte and application thereof | |
| CN111117910B (en) | Enterobacter ludwigii PN6 and application thereof | |
| CN103146610B (en) | Plant growth-promoting rhizobacteria and application thereof | |
| CN112358974B (en) | Plant endophytic fungus epicoccum nigrum FZT214 and application thereof | |
| CN111187737B (en) | Saline-alkali tolerant growth-promoting bacterium and application thereof in saline-alkali soil improvement | |
| CN104531546A (en) | Pseudomonas putida SRPG-396 and salt-dissolving and growth-promoting application thereof | |
| CN107502572A (en) | Application of the bacillus licheniformis in straw degradative, the microbial bacterial agent comprising the bacterium and its application | |
| CN103740634B (en) | A kind of Trichoderma asperellum chlamydospore pulvis and preparation method thereof and its purposes | |
| CN104263682B (en) | Plant-growth-promoting endophytic bacterium having polycyclic aromatic hydrocarbons degrading function and application thereof | |
| CN104531576A (en) | Strain for degrading dibutyl phthalate | |
| CN104371956A (en) | Bacillus with blocking effect on cadmium and application of bacillus | |
| CN110184225A (en) | One plant of rhizosphere growth-promoting endophytic bacteria PHE-2 and its application with PAHs degradation capability | |
| CN111117909B (en) | Strain capable of resisting multiple heavy metals and promoting plant growth and application thereof | |
| CN104450550A (en) | Screening method and application of Raoultella planticola strain SRPG-4 producing ACC (1-aminocyclopropane-1-carboxylic acid) deaminase activity | |
| CN114276950B (en) | Wide-pH-adaptability salt-tolerant phosphate solubilizing bacterium and application thereof | |
| CN109576177B (en) | Chinese micro-rod strain SM8 and application thereof in salt tolerance and growth promotion | |
| CN104630092A (en) | Tobacco growth-promoting rhizobacterium YC9 and application thereof | |
| CN103981103B (en) | DSE bacterial strain J-N3 and the application on dendrobium candidum is produced thereof | |
| CN111423995B (en) | Salt-tolerant growth-promoting effect of strain glutamicibacter soli1-3-3 and application thereof | |
| CN116121147B (en) | Chenopodium ambrosioides seed endophytic Larimol agrobacterium and application thereof | |
| CN114934000B (en) | Salt-tolerant bacillus amyloliquefaciens and screening and application thereof | |
| CN105274031A (en) | Sphingomonas sp. and its application | |
| CN103820359A (en) | Acquisition of bacteria for producing ACC deaminizing enzyme and promoting growth of ginseng and application thereof | |
| CN114806931A (en) | Bacillus belgii YQ-1-8 and application thereof | |
| 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 |
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 |