CN110982742A - Salt-tolerant growth-promoting bacteria wp-3 and application thereof - Google Patents
Salt-tolerant growth-promoting bacteria wp-3 and application thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of agricultural microorganisms and discloses a salt-tolerant growth-promoting bacterium wp-3 and application thereof, wherein the salt-tolerant growth-promoting bacterium wp-3 is classified and named as staphylococcus, and the gene sequence is SEQ ID NO: 1; and storing in a China center for typical culture preservation in 2018, 5 months and 14 days, wherein the preservation number is CCTCC NO: m2018273; the morphological characteristics of the salt-tolerant growth-promoting bacteria wp-3 are as follows: the colony is small, light yellow, opaque, smooth in surface, smooth in edge and round. The salt-tolerant growth-promoting bacteria wp-3 have higher alkali tolerance, salt tolerance and better adaptability to temperature; can dissolve organic phosphorus and inorganic phosphorus, increase the content of soluble phosphorus in soil, synthesize auxin IAA, promote the growth of crops and promote the germination of crop seeds.
Description
Technical Field
The invention belongs to the technical field of agricultural microorganisms, and particularly relates to salt-tolerant growth-promoting bacteria wp-3 and application thereof.
Background
Currently, the closest prior art: china has abundant land resources and large area, but saline-alkali soil and cultivated land have heavier salinization proportion. Some of these soils are too saline and alkaline to form a special environment and thus are difficult to grow. The special environment inoculates saline-alkali microorganisms adapting to the environment, particularly, the microorganism resources covered by vegetation in the special environment are richer, and the special environment has diversified genes, structures and functions. For example, some strains have the capacities of fixing nitrogen, dissolving phosphorus (inorganic phosphorus and organic phosphorus) and secreting plant hormones besides the saline-alkali resistance. Many studies have demonstrated that these strains with multiple functions are capable of promoting the growth of crops.
Currently, the current practice is. China lacks salt-tolerant growth-promoting microorganisms, especially high-salinity-tolerant growth-promoting microorganisms. Therefore, it is a hot spot of current research to excavate high-salt-tolerance growth-promoting microorganisms from saline-alkali soil, improve severe saline-alkali soil and promote crop growth by using the excavated high-salt-tolerance growth-promoting microorganisms.
In summary, the problems of the prior art are as follows: the prior art has less research on the salt-tolerant growth-promoting microorganisms, and lacks of the salt-tolerant growth-promoting microorganisms, particularly lacks of effective high-salt-tolerant growth-promoting microorganisms. The salt-tolerant growth-promoting bacteria wp-3 can survive in high-salinity soil, have better growth-promoting characteristics and promote the growth of crops.
The difficulty of solving the technical problems is as follows: the saline-alkali soil excavation can not only resist high salt, but also promote the growth of crops.
The significance of solving the technical problems is as follows: the saline-alkali soil is excavated with high salt tolerance and growth promoting microorganisms, and the excavated high salt tolerance growth promoting microorganisms can improve the severe saline-alkali soil, promote the growth of crops and provide strain resources for promoting the growth of crops in adverse circumstances.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a salt-tolerant growth-promoting bacterium wp-3 and application thereof.
The salt-tolerant growth-promoting bacteria wp-3 are classified and named as staphylococcus (Marinococcus sp.), and the gene sequence is SEQ ID NO: 1; and storing in a China center for typical culture preservation in 2018, 5 months and 14 days, wherein the preservation number is CCTCC NO: m2018273, preservation unit Address Wuhan city Wuchang district No. 299, Hubei province.
Another object of the invention is to provide a method for separating salt-tolerant growth-promoting bacteria wp-3, which comprises the following steps:
step one, collecting soil from 146 th group saline-alkali soil of Xinjiang Uyghur, filling the soil into a clean sampling bag, marking the soil, and storing the soil in a refrigerator at 4 ℃ for later use.
Step two, weighing 10g of the collected soil sample, placing the weighed soil sample into a 250m L triangular flask containing 90m L sterile water, shaking the soil sample on a shaking table for 20min at a speed of 150r/min, sucking 1ml of the supernatant by using a 1ml pipette, adding the supernatant into a test tube containing 9ml of sterile water, and fully and uniformly mixing.
Step three, sucking 1ml of the mixture from the test tube by a pipette, adding the mixture into another test tube containing 9ml of sterile water, uniformly mixing, and the like to obtain 10-1，10-2，10-3，10-4，10-5，10-6Solutions of different dilutions.
Step four, sucking 0.1ml of each solution with different concentrations obtained in the step three, respectively and uniformly coating the solution on an LB medium plate containing 10% of sodium chloride, and inversely culturing the solution in a 30 ℃ incubator for 2-3 days.
And fifthly, selecting a single colony to be inoculated on an LB inclined plane, and storing the single colony in a refrigerator at 4 ℃ after the lawn grows out.
Further, the LB medium is composed of 5g of yeast extract, 10g of tryptone, 10g of sodium chloride, 1000mL of distilled water, and 15g of solid agar; and sterilizing at 121 deg.C for 20 min.
Another object of the invention is to provide a method for identifying salt-tolerant growth-promoting bacteria wp-3, which comprises the following steps: streaking the strain wp-3 obtained after separation and purification on an LB culture medium for colony morphology observation, wherein the morphological characteristics of the salt-tolerant growth-promoting bacteria wp-3 are as follows: the bacterial colony is small, light yellow, opaque, smooth in surface, smooth in edge and round; and (3) analyzing the sequence of 16S rRNA of the separated and purified strain wp-3, wherein the strain is Marinococcus sp.
The invention also aims to provide application of the salt-tolerant growth-promoting bacteria wp-3 in severe saline-alkali soil, wherein the salt-tolerant growth-promoting bacteria wp-3 has high alkali tolerance, salt tolerance and good adaptability to temperature.
Another purpose of the invention is to provide an application of the salt-tolerant growth-promoting bacteria wp-3 in increasing the content of soluble phosphorus in soil, wherein the salt-tolerant growth-promoting bacteria wp-3 can dissolve organic phosphorus and inorganic phosphorus.
Further, the quantitative determination method for the phosphorus dissolving capacity of the strain wp-3 comprises the following steps:
(1) 2ug m L-1The phosphorus standard solution is diluted in a gradient way until the phosphorus content is 0.00, 0.04, 0.08, 0.24, 0.40, 0.80 and 1.20ug m L-1A standard curve was prepared.
(2) Adding molybdenum antimony color-developing agent at room temperature, developing for 20min, measuring light absorption value and making standard curve.
(3) Selecting a little precipitate of the preserved phosphorus-dissolving strain, inoculating in LB liquid culture medium, and culturing at 30 deg.C for 150 r.min-1Shaking for 24h, changing the absorbance value OD600 of the bacterial liquid to 1, and sucking 1mL of bacterial liquid into a liquid PKO culture medium.
(4) Selecting a little precipitate of phosphate-solubilizing strain, inoculating in LB liquid culture medium, and culturing at 30 deg.C for 150r min-1The cells were cultured overnight with shaking, and the absorbance of the cells was 1 at OD 600.
(5) Sucking 1m L bacterial liquid into liquid Monkina culture medium, and culturing at 30 deg.C for 120r min-1The culture was performed in a shaking incubator for 48 h.
(6) Taking 5m L bacterial liquid, centrifuging, adding color-developing agent after the supernatant reaches 25m L constant volume, wherein 3 groups are parallel, and measuring the phosphorus dissolving and dissolving capacity of the strain by a molybdenum-antimony colorimetric method.
Further, the Monkina organic medium (1L) was composed of 10.0g of glucose, (NH)4)2SO40.5 g，MgSO4·7H2O 0.3g，NaCl 0.3g，KCl 0.3g，FeSO4·7H2O 0.03g，MnSO4·H20.03g of O, 0.2g of lecithin, CaCO31.0 g, 0.5g of yeast powder and 20.0g of agar; 1000ml of distilled water; the liquid medium was sterilized at 121 ℃ for 20 minutes without agar.
The PKO inorganic medium (1L) was composed of glucose 10.0g, MgSO4·7H2O 0.3g，(NH4)2SO40.5 g，NaCl 0.3g，Ca3(PO4)22.0g，KCl 0.3g，MnSO4·H2O 0.03g，FeSO4·7H20.036g of O, 20g of agar and 1000ml of distilled water; the liquid medium was sterilized at 121 ℃ for 20 minutes without agar.
The invention also aims to provide application of the halophilic growth-promoting bacteria wp-3 in synthesis of auxin IAA, and the halophilic growth-promoting bacteria wp-3 can synthesize the auxin IAA by using tryptophan.
Further, the method for measuring the capability of the strain wp-3 for secreting IAA comprises the following steps:
IAA standard curve configuration
32.8mg of IAA is accurately weighed in a 100m L volumetric flask, methanol is used for constant volume (the solution contains IAA328mg/L, methanol is used for sequentially diluting for 1 time and continuously carrying out for 4 times to prepare 5 parts of standard solutions with the concentration sequentially different by 1 time, and the standard solutions with the same amount are added with the colorimetric solution with the same amount during colorimetric process.
(II) inoculating the strain wp-3 in an LB liquid medium containing L-tryptophan (100mg/L) according to the inoculation amount of 1%, and culturing for 48h at 30 ℃ and 150r/min in a shaking way.
(III) centrifuging the bacterial suspension at 10000r/min for 10min, taking the supernatant, adding an equal volume of Salkowski colorimetric solution (50mL of 35% HClO)4+1m L 0.5mol/L FeCl3) And kept standing for 30min in the dark, and the OD530 value is measured.
(IV) calculating the IAA content in the culture solution per unit volume by contrasting with a standard curve.
The invention also aims to provide application of the salt-tolerant growth-promoting bacteria wp-3 in promoting crop growth, and the salt-tolerant growth-promoting bacteria wp-3 can promote the growth of wheat with high salinity stress.
Further, the determination method for promoting the growth of the wheat with high salinity stress by the salt-tolerant growth-promoting bacteria wp-3 comprises the following steps:
1) 2 treatments are set: t1: 150mM NaCl solution; t2: 150mM NaCl solution + bacterial suspension; each treatment is provided with 3 repetitions, and 3 wheat seeds are placed in each repeated flowerpot.
2) The wheat seeds are washed clean by tap water, sterilized by 75% alcohol for 30s, sterilized by 0.1% mercuric chloride for 7min and washed by sterile water for 5-6 times.
3) Soaking the seeds in sterile water for 2h, soaking the wheat seeds in each treatment solution for 8h, taking the plump seeds with consistent sizes, and uniformly arranging the seeds in a sterilized germination box paved with sterilized filter paper and having the diameter of 12 cm.
4) After wheat seeds are whitened, the wheat seeds are immediately sowed in nutrient soil (nutrient soil 3: vermiculite 1) in a flowerpot (the upper diameter is 15cm, the lower diameter is 10cm, the height is 13cm), the sowing depth is about 1cm, and 1-time uniform sowing is carried out on a sowing belt by using gloves; watering in time with equal amount, and measuring growth index after 50 days.
Another purpose of the invention is to provide an application of the salt-tolerant growth-promoting bacteria wp-3 in promoting the germination of crop seeds, wherein the salt-tolerant growth-promoting bacteria wp-3 can promote the germination of wheat seeds with salt stress.
Further, the determination method for promoting salt stress wheat seed germination by the salt-tolerant growth-promoting bacteria wp-3 comprises the following steps:
① sets 2 treatments (T1: 150mM NaCl solution; T2: 150mM NaCl solution + strain suspension.) 3 replicates per treatment were set, and 48 wheat seeds were placed in each replicate germination box.
② the wheat seeds are washed clean with tap water, sterilized with 75% alcohol for 30s, sterilized with 0.1% mercuric chloride for 7min, and washed with sterile water for 5-6 times.
③ soaking the seeds in sterile water for 2 hr, soaking the wheat seeds in the solutions for 8 hr, uniformly arranging the plump seeds in a sterilized germination box with sterilized filter paper and diameter of 12cm, and calculating the germination rate of wheat every 24 hr after 48 hr.
In summary, the advantages and positive effects of the invention are: the strain can effectively promote the absorption of plant nutrition, regulate the growth of plants and improve the stress resistance of plants under the stress condition.
The salt-tolerant growth-promoting bacteria wp-3 and the application thereof provided by the invention have the advantages that the salt-tolerant growth-promoting bacteria wp-3 have high alkali tolerance, salt tolerance and good adaptability to temperature; can dissolve organic phosphorus and inorganic phosphorus, increase the content of soluble phosphorus in soil, synthesize auxin IAA, promote the growth of crops and promote the germination of crop seeds.
Drawings
FIG. 1 is a flow chart of a method for isolating strain wp-3 according to the example of the present invention.
Fig. 2 is a schematic diagram of a colony morphology of a high salt tolerant growth-promoting bacterium (Marinococcus sp.) provided by an embodiment of the present invention.
FIG. 3 is a diagram of the phosphorus-tolerant ability of strain wp-3 provided by the example of the present invention.
FIG. 4 is a diagram showing a quantitative determination standard curve of strain wp-3IAA provided in the example of the present invention.
FIG. 5 is a schematic diagram of the growth promoting effect of a wheat inoculation strain wp-3 provided by the embodiment of the invention.
FIG. 6 is a schematic diagram showing the effect of a wheat inoculum strain wp-3 provided by an example of the present invention on root length, fresh weight of seedlings, fresh weight of above-ground and below-ground parts.
FIG. 7 is a schematic diagram showing the growth promoting effect of a wheat inoculation strain wp-3 under salt stress provided by the embodiment of the invention.
FIG. 8 is a graph showing the effect of salt stress wheat inoculum strain wp-3 on root length, fresh weight of seedlings, fresh weight of above-ground and below-ground parts.
FIG. 9 is a schematic diagram showing the effect of salt stress wheat seed inoculum strain wp-3 on germination percentage according to an embodiment of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The prior art has less research on the salt-tolerant growth-promoting microorganisms, and lacks of the salt-tolerant growth-promoting microorganisms, particularly lacks of effective high-salt-tolerant growth-promoting microorganisms.
Aiming at the problems in the prior art, the invention provides a salt-tolerant growth-promoting bacterium wp-3 and application thereof, and the invention is described in detail with reference to the accompanying drawings.
The salt-tolerant growth-promoting bacteria wp-3 provided by the embodiment of the invention is classified and named as staphylococcus (Marinococcus), and the gene sequence is SEQ ID NO: 1; and storing in a China center for typical culture preservation in 2018, 5 months and 14 days, wherein the preservation number is CCTCC NO: m2018273, preservation unit Address Wuhan city Wuchang district No. 299, Hubei province.
As shown in FIG. 2, the morphological characteristics of the salt tolerant growth-promoting bacteria wp-3 provided by the embodiment of the invention are as follows: the colony is small, light yellow, opaque, smooth in surface, smooth in edge and round.
The application of the salt-tolerant growth-promoting bacteria wp-3 provided by the embodiment of the invention in severe saline-alkali soil, wherein the salt-tolerant growth-promoting bacteria wp-3 have high alkali tolerance, salt tolerance and good adaptability to temperature.
The salt-tolerant growth-promoting bacteria wp-3 provided by the embodiment of the invention can dissolve organic phosphorus and inorganic phosphorus in the soil.
The salt-tolerant growth-promoting bacteria wp-3 provided by the embodiment of the invention can be used for synthesizing auxin IAA by using tryptophan.
The embodiment of the invention provides application of the salt-tolerant growth-promoting bacteria wp-3 in promoting crop growth, and the salt-tolerant growth-promoting bacteria wp-3 can promote the growth of wheat with high salinity stress.
The salt-tolerant growth-promoting bacteria wp-3 provided by the embodiment of the invention can promote germination of salt-stressed wheat seeds.
The present invention will be further described with reference to the following examples.
Example 1: separation and identification of high-salt-tolerance growth-promoting bacteria wp-3(Marinococcus sp.)
1.1 isolation of high salt tolerant growth-promoting bacteria wp-3(Marinococcus sp.) (as shown in FIG. 1)
Soil was collected from 146 th group saline and alkaline land of Xinjiang Shi, and the soil was put into a clean sampling bag, marked and stored in a refrigerator at 4 ℃ for future use. Weighing 10g of the collected soil sample, placing the soil sample into a 250m L triangular flask containing 90m L sterile water, shaking the soil sample in a shaking table for 20min at a speed of 150r/min, sucking 1ml of the soil sample from the supernatant by using a 1ml pipette, adding the soil sample into a test tube containing 9ml of sterile water, fully and uniformly mixing, sucking 1ml of the soil sample from the test tube by using a pipette, adding the soil sample into another test tube containing 9ml of sterile water, uniformly mixing, and so on to obtain 10g of the soil sample-1，10-2，10-3，10-4，10-5，10-6After the solutions with different dilutions, 0.1ml of each solution is absorbed and respectively and uniformly coated on an LB medium plate containing 10% sodium chloride, the solution is inverted and cultured in a constant temperature box at 30 ℃ for 2-3d, a single colony is selected to be inoculated on an LB inclined plane, and the solution is stored in a refrigerator at 4 ℃ after a lawn grows out.
LB culture medium: 5g of yeast extract, 10g of tryptone, 10g of sodium chloride, 1000mL of distilled water, 15g of solid agar and sterilization at 121 ℃ for 20 minutes.
1.2 identification of Strain wp-3
The isolated and purified strain wp-3 was streaked on LB medium to observe the colony morphology, as shown in FIG. 1. It features small bacterial colony, light yellow, opaque, smooth surface, smooth edge and circular shape.
And (3) analyzing the sequence of 16S rRNA of the separated and purified strain wp-3, wherein the strain is Marinococcus.
Example 2: study on growth characteristics of Strain wp-3
2.1 different pH effects on growth of Strain wp-3
The bacterial suspension of the bacterial strain wp-3 is inoculated in LB culture media with different pH values (6, 7, 8, 9, 10, 11) in an inoculation amount of 1.0 percent, the inoculated bacterial suspension is cultured for 24 hours at 30 ℃ under the shaking condition of 150r/min, the LB culture medium without inoculation is used as a reference, a light density value (D600 nm) is detected by adopting a 722 spectrophotometer, and the result is shown in Table 1, and the optimal growth pH is 9.
TABLE 1 Effect of different pH on growth of strain wp-3
2.2 different temperatures influence the growth of the strain wp-3
Inoculating 1.0% strain wp-3 suspension into LB medium with pH of 9, culturing at different temperatures (24 deg.C, 27 deg.C, 30 deg.C, 33 deg.C, 36 deg.C) under shaking at 150r/min for 24h, detecting optical density value (D600 nm) with 722 spectrophotometer with reference to the uninoculated LB medium, and finding out the result in Table 2 with optimum growth temperature of 30 deg.C.
TABLE 2 Effect of different temperatures on the growth of strain wp-3
2.3 Effect of different salinity on growth of Strain wp-3
The strain wp-3 suspension is inoculated in LB culture medium with pH value of 9 and different sodium chloride concentrations (4-17%) in an inoculation amount of 1.0%, shaking culture is carried out for 24h at 30 ℃ and 150r/min, a light density value (D600 nm) is detected by adopting a 722 spectrophotometer by taking the uninoculated LB culture medium as a reference, and the result is shown in Table 3, and the maximum salt tolerance concentration of the strain is 17%.
TABLE 3 Effect of different salinity on growth of strain wp-3
Example 3: quantitative determination of phosphorus dissolving capacity of strain wp-3
3.1 mixing 2ug m L-1The phosphorus standard solution is diluted in a gradient way until the phosphorus content is 0.00, 0.04, 0.08, 0.24, 0.40, 0.80 and 1.20ug m L-1A standard curve was prepared. Adding molybdenum antimony at room temperatureAnd (5) developing for 20min by using an anti-color-developing agent, measuring the light absorption value of the anti-color-developing agent, and making a standard curve. Selecting a little precipitate of the preserved phosphorus-dissolving strain, inoculating in LB liquid culture medium, and culturing at 30 deg.C for 150 r.min-1Performing shake culture for 24h, enabling the light absorption value OD600 of the bacterial liquid to be 1, and sucking 1mL of bacterial liquid into a liquid PKO culture medium; selecting a little precipitate of phosphate-solubilizing strain, inoculating in LB liquid culture medium, and culturing at 30 deg.C for 150r min-1Shaking for overnight culture to make the absorbance OD600 ═ 1, sucking 1m L bacterial liquid into Monkina liquid culture medium, and culturing at 30 deg.C for 120r min-1The culture was performed in a shaking incubator for 48 h. Taking 5m L bacterial liquid, centrifuging, adding color-developing agent after the supernatant reaches 25m L constant volume, wherein 3 groups are parallel, and measuring the phosphorus dissolving and dissolving capacity of the strain by a molybdenum-antimony colorimetric method.
The strain wp-3 can dissolve organic phosphorus, and the transformation amount is 0.22 mg/L2 d; inorganic phosphorus was dissolved, and the conversion amount was 1.84 mg/L2 d, as shown in FIG. 3.
Monkina organic medium (1L): glucose 10.0g, (NH)4)2SO40.5 g，MgSO4·7H2O 0.3g，NaCl0.3g，KCl 0.3g，FeSO4·7H2O 0.03g，MnSO4·H20.03g of O, 0.2g of lecithin, CaCO31.0 g, 0.5g of yeast powder and 20.0g of agar; 1000ml of distilled water; the liquid medium was sterilized at 121 ℃ for 20 minutes without agar.
PKO inorganic medium (1L): glucose 10.0g, MgSO4·7H2O 0.3g，(NH4)2SO40.5g，NaCl0.3g，Ca3(PO4)22.0g，KCl 0.3g，MnSO4·H2O 0.03g，FeSO4·7H20.036g of O, 20g of agar and 1000ml of distilled water; the liquid medium was sterilized at 121 ℃ for 20 minutes without agar.
Example 4: quantitative determination of IAA yield of strain wp-3
4.1IAA Standard Curve configuration
32.8mg of IAA is accurately weighed in a 100m L volumetric flask, methanol is used for constant volume (the solution contains IAA328mg/L, methanol is used for sequentially diluting for 1 time and continuously carrying out for 4 times to prepare 5 parts of standard solutions with the concentration sequentially different by 1 time, the standard solutions are equal in quantity and the colorimetric solution is equal in quantity in colorimetric process, and the standard curve is shown in figure 4.
4.2 measurement of IAA secretion ability of Strain wp-3
Inoculating strain wp-3 at 1% inoculum size in LB liquid culture medium containing L-tryptophan (100mg/L), shake culturing at 30 deg.C and 150r/min for 48 hr, centrifuging the bacterial suspension at 10000r/min for 10min, collecting supernatant, adding equal volume Salkowski colorimetric solution (50mL 35% HClO)4+1m L 0.5mol/L FeCl3) And kept standing for 30min in the dark, and the OD530 value is measured. The IAA content per volume of the culture broth was calculated against a standard curve. The IAA content of the strain wp-3 is determined to be 21.16 mg/L.
Example 5: growth-promoting potting experiment of strain wp-3 on salt-stress-free wheat
5.1 wheat potting experiment
2 treatments were set (T1: sterile water; T2: sterile water + strain suspension). Each treatment is provided with 3 repetitions, and 3 wheat seeds are placed in each repeated flowerpot. The wheat seeds are washed clean by tap water, sterilized by 75% alcohol for 30s, sterilized by 0.1% mercuric chloride for 7min, washed by sterile water for 5-6 times, soaked in the sterile water for 2h, taken and soaked in each treatment solution for 8h, and then uniformly arranged in a sterilized germination box (diameter of 12cm) paved with sterilized filter paper. After wheat seeds are exposed to white, immediately sowing the wheat seeds into a flowerpot (the upper diameter is 15cm, the lower diameter is 10cm and the height is 13cm) filled with sterilized nutrient soil (nutrient soil 3: vermiculite 1) with the same quantity, wherein the sowing depth is about 1cm, and 1-time uniform sowing is carried out on the sowing belt by using gloves. Watering in time with equal amount, and measuring growth index after 50 days. The growth promoting effect is shown in figure 5.
5.2 wheat growth index determination
The wheat grows for 50 days, a plant is taken out, the root is cleaned, the root length, the fresh weight of seedlings, the fresh weight of overground parts and the fresh weight of underground parts of the wheat are measured, the strain wp-3 has the promoting effect on the root length, the fresh weight of seedlings, the fresh weight of overground parts and the fresh weight of underground parts of the wheat, the root length, the fresh weight of the overground parts and the fresh weight of the underground parts are respectively increased by 25.5%, 13.79%, 184.2% and 19.0%, and the result is shown in figure 6.
Example 6: growth-promoting potting experiment of strain wp-3 on salt-stressed wheat
6.1 wheat potting experiment
2 treatments were set (T1: 150mM NaCl solution; T2: 150mM NaCl solution + strain suspension). Each treatment is provided with 3 repetitions, and 3 wheat seeds are placed in each repeated flowerpot. The wheat seeds are washed clean by tap water, sterilized by 75% alcohol for 30s, sterilized by 0.1% mercuric chloride for 7min, washed by sterile water for 5-6 times, soaked in the sterile water for 2h, soaked in each treatment solution for 8h, taken as plump seeds with consistent sizes, and then uniformly arranged in a sterilized germination box (the diameter is 12cm) paved with sterilized filter paper. After wheat seeds are exposed to white, immediately sowing the wheat seeds into a flowerpot (the upper diameter is 15cm, the lower diameter is 10cm and the height is 13cm) filled with sterilized nutrient soil (nutrient soil 3: vermiculite 1) with the same quantity, wherein the sowing depth is about 1cm, and 1-time uniform sowing is carried out on the sowing belt by using gloves. Timely watering in equal amount, and measuring growth indexes after 50 days, wherein the growth promoting effect is shown in figure 7.
6.2 measurement of physiological index of wheat growth
The wheat grows for 50 days, the plant is taken out, the root is cleaned, the root length, the fresh weight of seedlings, the fresh weight of overground parts and the fresh weight of underground parts of the wheat are measured, the strain wp-3 has the promoting effect on the root length, the fresh weight of seedlings and the fresh weight of the overground parts of the wheat, the root length, the fresh weight of the overground parts of the wheat are respectively increased by 12.8%, 25.8% and 11.9%, and the result is shown in figure 8.
Example 7: wheat inoculation strain wp-3 germination experiment under salt stress
2 treatments were set (T1: 150mM NaCl solution; T2: 150mM NaCl solution + strain suspension). Each treatment was set to 3 replicates and 48 wheat seeds were placed in each replicate germination box. The wheat seeds are washed clean by tap water, sterilized by 75% alcohol for 30s, sterilized by 0.1% mercuric chloride for 7min, washed by sterile water for 5-6 times, soaked in the sterile water for 2h, soaked in each treatment solution for 8h, taken as plump seeds with consistent sizes, and then uniformly arranged in a sterilized germination box (the diameter is 12cm) paved with sterilized filter paper. After 48h, the germination rate of wheat is calculated every 24h, and the germination rate of wheat is higher than that of the control in 48h to 168h, as shown in figure 9.
Proof part (concrete examples/experiments/simulation/pharmacological analysis/positive experimental data, evidential material, appraisal reports, business data, research and development evidence, business cooperation evidence, etc. capable of proving the inventive aspects of the present invention)
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Sequence listing
<110> river university
<120> salt-tolerant growth-promoting bacteria wp-3 and application
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<170>SIPOSequenceListing 1.0
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<211>1185
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
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aatctgtcac ttcggcggct ccaagggtta ccgacttcgg gtgttgcaaa ctctcgtggt 60
gtgacgggcg gtgtgtacaa ggcccgggaa cgtattcacc gtggcatgct gatccacgat 120
tactagcgat tccggcttca tgcaggcgag ttgcagcctg caatccgaac tgagaatggc 180
tttgcgagat tggctccacc tcgcggtttc gcatccctct gtaccatcca ttgtagcacg 240
tgtgtagccc aggtcataag gggcatgatg atttgacgtc gtccccacct tcctccggtt 300
tgtcaccggc agtcacctta gagtgcccaa ctgaatgctg gcaactaaga ttaagggttg 360
cgctcgttgc gggacttaac ccaacatctc acgacacgag ctgacgacaa ccatgcacca 420
cctgtcattt tgtcccccga aggggaacct tctgtctcca gaagtagcaa aagatgtcaa 480
gacctggtaa ggttcttcgc gttgcgtcga attaaaccac atgctccacc gcttgtgcgg 540
gcccccgtca attcttttga gtttcagcct tgcggccgta ctccccaggc ggagtgctta 600
atgcgttagc ttcggcacta agggcatcga aacccctaac acctagcact catcgtttac 660
ggcgtggact accagggtat ctaatcctgt ttgctcccca cgctttcgcg cctcagcgtc 720
aataacagac cagagagccg ccttcgccac tggtgttcct ccacatatct acgcatttca 780
ccgctacacg tggaattcca ctctcctctt ctgcattcca gttctccagt ttccaatgac 840
cctccacggt tgagccgtgg gctttcacat cagacttaaa gaaccgcctg cgcgcgcttt 900
acgcccaata attcccggac aacgcttgcc ccctacgtat taccgcggct gctggcacgt 960
agttagcccg gggctttctg gtgaggtacc gtcagggggc ggccctgtta gaaccgccct 1020
tgttcttccc tccacaacag agctttacga tccgaaaacc ttcatcacct cacgcggcgt 1080
gcaccgtcag actttcgtca ttgcgatgat cccttactgc ctggcctccc ggtagagtct 1140
ggcgtgttct cagtccagtg tggcccgatc caccccctct ctcaa 1185
Claims (10)
1. The salt-tolerant growth-promoting bacteria wp-3, which are preserved in a China typical culture preservation center in 2018, 5 months and 14 days, and the preservation number is CCTCC NO: m2018273, preservation unit Address Wuhan city Wuchang district No. 299, Hubei province.
2. The salt-tolerant growth-promoting bacteria wp-3 of claim 1, wherein said salt-tolerant growth-promoting bacteria wp-3 DNA gene sequence is SEQ ID NO: 1.
3. the method for separating the salt-tolerant growth-promoting bacteria wp-3 according to claim 1, wherein the method for separating the salt-tolerant growth-promoting bacteria wp-3 comprises the following steps:
collecting soil from 146 th group saline-alkali soil of Xinjiang Ushi, filling the soil into a clean sampling bag, marking, and storing the soil in a refrigerator at 4 ℃ for later use;
step two, weighing 10g of the collected soil sample, placing the weighed soil sample into a 250m L triangular flask containing 90m L sterile water, shaking the soil sample in a shaking table for 20min at a speed of 150r/min, sucking 1ml of the supernatant by using a 1ml pipette, adding the supernatant into a test tube containing 9ml of sterile water, and fully and uniformly mixing the supernatant and the test tube;
step three, sucking 1ml from the test tube by a pipette, adding into another test tube containing 9ml of sterile water, mixing uniformly, and so on to obtain 10-1，10-2，10-3，10-4，10-5，10-6Solutions of different dilutions;
step four, sucking 0.1ml of each solution with different concentrations obtained in the step three, respectively and uniformly coating the solution on an LB medium plate containing 10% of sodium chloride, and inversely culturing the solution in a 30 ℃ incubator for 2-3 days;
and fifthly, selecting a single colony to be inoculated on an LB inclined plane, and storing the single colony in a refrigerator at 4 ℃ after the lawn grows out.
4. The method for separating halophythmic-promoting bacteria wp-3 of claim 3, wherein said LB medium consists of yeast extract 5g, tryptone 10g, sodium chloride 10g, distilled water 1000mL, solids plus agar 15 g; and sterilizing at 121 deg.C for 20 min.
5. The method for identifying the salt-tolerant growth-promoting bacteria wp-3 according to claim 1, wherein the method for identifying the salt-tolerant growth-promoting bacteria wp-3 comprises:
streaking the separated and purified strain wp-3 on an LB culture medium for colony morphology analysis;
the isolated and purified strain wp-3 was analyzed by 16S rRNA sequencing.
6. The application of the salt-tolerant growth-promoting bacteria wp-3 as claimed in claim 1 in severe saline-alkali soil.
7. Use of the salt-tolerant growth-promoting bacteria wp-3 of claim 1 for increasing the soluble phosphorus content of soil.
8. Use of the halophythmogenic bacteria wp-3 of claim 1 in the synthesis of auxin IAA.
9. Use of the salt-tolerant growth-promoting bacteria wp-3 of claim 1 for promoting crop growth.
10. Use of the salt-tolerant growth-promoting bacteria wp-3 of claim 1 for promoting germination of crop seeds.
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101457213A (en) * | 2009-01-06 | 2009-06-17 | 东北农业大学 | Marinococcus and its application in degrading imazethapyr |
| CN101597581A (en) * | 2009-07-13 | 2009-12-09 | 华北电力大学 | A kind of Alkaliphilic bacillus and cultural method thereof and application |
| CN103097538A (en) * | 2009-12-22 | 2013-05-08 | 宝洁公司 | Branched-chain fatty acids and biological production thereof |
| CN107746822A (en) * | 2017-11-24 | 2018-03-02 | 陕西省微生物研究所 | Salt affected soil repairs microbial bacterial agent and preparation method thereof |
| US20190264232A1 (en) * | 2018-02-23 | 2019-08-29 | Pioneer Hi-Bred International, Inc. | Novel cas9 orthologs |
| CN110607263A (en) * | 2019-09-26 | 2019-12-24 | 石河子大学 | Bacillus alcalophilus strain wp-1 and application thereof |
| CN110622996A (en) * | 2019-09-26 | 2019-12-31 | 江苏国信协联能源有限公司 | Microbial agent and application thereof |
| CN111039726A (en) * | 2019-12-10 | 2020-04-21 | 石河子大学 | Microbial fertilizer for improving saline-alkali soil, preparation method and application |
-
2019
- 2019-12-12 CN CN201911270615.5A patent/CN110982742B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101457213A (en) * | 2009-01-06 | 2009-06-17 | 东北农业大学 | Marinococcus and its application in degrading imazethapyr |
| CN101597581A (en) * | 2009-07-13 | 2009-12-09 | 华北电力大学 | A kind of Alkaliphilic bacillus and cultural method thereof and application |
| CN103097538A (en) * | 2009-12-22 | 2013-05-08 | 宝洁公司 | Branched-chain fatty acids and biological production thereof |
| CN107746822A (en) * | 2017-11-24 | 2018-03-02 | 陕西省微生物研究所 | Salt affected soil repairs microbial bacterial agent and preparation method thereof |
| US20190264232A1 (en) * | 2018-02-23 | 2019-08-29 | Pioneer Hi-Bred International, Inc. | Novel cas9 orthologs |
| CN110607263A (en) * | 2019-09-26 | 2019-12-24 | 石河子大学 | Bacillus alcalophilus strain wp-1 and application thereof |
| CN110622996A (en) * | 2019-09-26 | 2019-12-31 | 江苏国信协联能源有限公司 | Microbial agent and application thereof |
| CN111039726A (en) * | 2019-12-10 | 2020-04-21 | 石河子大学 | Microbial fertilizer for improving saline-alkali soil, preparation method and application |
Non-Patent Citations (4)
| Title |
|---|
| FRANCESCA MAPELLI等: "Potential for Plant Growth Promotion of Rhizobacteria Associated with Salicornia Growing in Tunisian Hypersaline Soils", 《BIOMED RESEARCH INTERNATIONAL》 * |
| SANJAY ARORA等: "Microbial Approach for Bioremediation of Saline and Sodic Soils", 《BIOREMEDIATION OF SALT AFFECTED SOILS》 * |
| WEN-JUN LI等: "Marinococcus halotolerans sp. nov., isolated from Qinghai, north-west China", 《INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY》 * |
| 赵建朋: "新疆艾比湖中度嗜盐菌生物学特性研究和耐盐基因的克隆及分析", 《中国优秀硕士学位论文全文数据库（电子期刊）基础科学辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112680386A (en) * | 2021-02-03 | 2021-04-20 | 河南大学 | Application of corn rhizosphere growth-promoting bacteria in promoting plant growth |
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