CN113481117A - Multifunctional bacterium C1 for improving soda saline-alkali soil and application thereof - Google Patents
Multifunctional bacterium C1 for improving soda saline-alkali soil and application thereof Download PDFInfo
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Abstract
The strain C1 is preserved in the common microorganism center of China general microbiological culture Collection center, is proposed to be classified and named as Paenibacilus sp, has the preservation number of CGMCC No.20154, and has the preservation time of 28 days 6 and 28 days 2020; the bacterium C1 is salt-alkali resistant, the pH is reduced by using glucose to produce organic acid, the rice straw is degraded to produce cellulase, exopolysaccharide is secreted, the bacterium solution, the rice straw, ammonium sulfate and calcium phosphate are applied to soil to promote the degradation of the rice straw, the total amount of organic carbon and the water steady-state content in the soil are increased, the formation of macro-aggregates and water-stable aggregates is promoted, the conductivity is reduced, the growth of a plant root system is promoted, the absorption capacity of plants on phosphorus is improved, and the problems of slow speed and single effect in the process of improving soda saline-alkali soil by the rice straw are solved.
Description
Technical Field
The invention relates to the field of agricultural microorganism application, in particular to a multifunctional bacterium C1 for improving soda saline-alkali soil and application thereof.
Background
The soda saline-alkali soil in China is mainly distributed in regions such as northeast Songxing plain, and the pH value of the soil is higher due to higher salt content and alkalization degree of the soil, the bioavailability of phosphorus in the soil is lower, and the formation and holding capacity of soil organic matters are lower, so that the quantity and functions of soil microorganisms are influenced, and the natural recovery difficulty of the soda saline-alkali soil is high. The rice straw returning soil can increase the organic matter content of the soil, improve the soil structure and the water permeability and air permeability, and has good application prospect in the improvement of soda saline-alkali soil, but because the soda saline-alkali soil in China is mainly in northeast regions with longer low temperature period, the soil microorganism quantity is small, the plant nutrient structure is unreasonable, the conversion rate of the rice straw in the soil is low, the formed organic matter is easy to lose alkali solution, and the improvement effect is influenced, so that the improvement of the soda saline-alkali soil is promoted by using the saline-alkali tolerant indigenous microorganisms and the nutrient substances necessary for the growth of the indigenous microorganisms.
The microorganisms mainly comprise bacteria for producing organic acid, degrading cellulose and producing extracellular secretion at present, not only are the conditions of lacking of strain resources, but also are mainly used for single-function microorganisms in application, multifunctional microorganism resources are lacked, the research on the soda saline-alkali soil improvement method is carried out, the application of the microorganisms in the soda saline-alkali soil improvement is severely restricted, and the strain resources for improving the soda saline-alkali soil of Songnen plain and the application thereof are rare, so that the multifunctional microorganisms suitable for the Songnen plain soda saline-alkali soil are urgently needed to be obtained, and the corresponding application method and technology are assisted to realize the rapid transformation of organic materials in the soda saline-alkali soil, the rapid optimization of the soil structure and the activation and maintenance of nutrient substances, so that the sustainable utilization requirement after the soda saline-alkali soil improvement is met.
Disclosure of Invention
The invention aims to provide a saline-alkali-resistant multifunctional strain named C1, which is used in the process of returning rice straws to the field of soda saline-alkali soil, can reduce the pH value of the soil and increase the quick-acting phosphorus content in the soil by generating organic acid, degrades the rice straws to generate cellulase and secrete extracellular polysaccharide, thereby accelerating the degradation rate of the rice straws in the soil, increasing the total amount of organic carbon and the water steady-state content in the soil, increasing the proportion of macro-aggregates and water-stable aggregates in the soil, realizing the simultaneous elimination of multiple barrier factors of the soda saline-alkali soil and promoting the sustainable utilization of the soda saline-alkali soil.
The multifunctional bacterium C1 for improving soda saline-alkali soil provided by the invention is deposited in the China general microbiological culture Collection center (address No. 3 of Xilu No. 1 of Kyoto-the-south district in Beijing), and is suggested to be classified and named as Paenibacillus sp (Paenibacillus sp.), the preservation number is CGMCC No.20154, and the preservation time is 28 days 6 months 6 years 2020.
The multifunctional bacterium C1 for improving soda saline-alkali soil is obtained by the following method;
step S1, adding 10g of fresh soda saline-alkali soil into 90mL of sterile water, oscillating for 30min at 28 ℃ and 160r/min, standing for 10min, adding 1mL of soil suspension into an improved LB liquid culture medium, culturing for 24h at 28 ℃ and 160r/min, coating the culture solution on a soil broth solid culture medium by a gradient dilution method, culturing for 48h at 28 ℃, and performing streak purification on single colonies with obviously different appearances by using the soil broth solid culture medium to obtain a purified strain;
the improved LB liquid culture medium is prepared by mixing 10g of tryptone, 30g of NaCl, 5g of yeast extract powder and 1L of distilled water, and adjusting the pH value to 8 by using 1mol/L of NaOH;
the soil broth solid culture medium is prepared by mixing 5g of glucose, 5g of calcium phosphate, 0.0584 g of monopotassium phosphate, 0.1547g of dipotassium phosphate, 15-18g of agar and 1000mL of soil leaching liquor, and adjusting the pH value to 7;
step S2, respectively taking 1 ring of the purified strain obtained in the step S1, inoculating the purified strain into 5mL of LB culture medium, oscillating the culture medium for 24 hours under the conditions of 28 ℃ and 160r/min, completely inoculating the culture into 100mL of fresh LB culture medium, culturing the culture medium for 24 hours under the same conditions to obtain fermentation liquor, centrifuging the fermentation liquor for 10 minutes under the conditions of 4 ℃ and 8000r/min, collecting bacterial cells, suspending the bacterial cells in sterile water, and preparing the bacterial cells with the number of viable bacteria not less than 109Inoculating the bacterial suspension of CFU/mL into an improved NBRIP culture medium according to the proportion of 2%, carrying out shake culture for 5d at the temperature of 28 ℃ and under the condition of 160r/min, and reserving a bacterial strain with the pH of fermentation liquor being less than 5.5;
the improved NBRIP culture medium comprises 10g of glucose, 5g of calcium phosphate, 0.5g of ammonium sulfate, 0.3g of magnesium sulfate heptahydrate, 0.03g of manganese sulfate heptahydrate, 0.03g of ferric sulfate heptahydrate, 0.3g of potassium chloride, 20g of sodium chloride, 18g of agar and 1000mL of distilled water which are mixed, and the pH value is 7;
and S3, preparing bacterial suspension from the strains retained in the step S3 according to the method in the step S2, inoculating the bacterial suspension into a rice straw degradation liquid culture medium according to a proportion of 2%, carrying out shaking culture at 28 ℃ for 7d, washing the residual rice straws in the fermentation liquid with sterile water for 2-3 times, then placing the washed residual rice straws in a 50 ℃ oven for drying, weighing, calculating the straw degradation rate, and retaining the strains with the rice straw degradation rate of more than 45%.
The rice straw degradation liquid culture medium comprises the following components: 1g of rice straw, 1g of potassium nitrate, 0.5g of ammonium sulfate, 0.5g of dipotassium phosphate, 0.5g of magnesium sulfate heptahydrate, 1.5g of sodium chloride, 15-18g of agar and 1000mL of distilled water, wherein the pH value is 7;
and S4, culturing the strain reserved in the step S3 by using a rice straw degradation liquid culture medium according to the method in the step S3, standing for 10min after the culture is finished, centrifuging the upper layer fermentation liquor at 8000r/min for 15min, mixing the supernatant with absolute ethyl alcohol according to the ratio of 1:3, standing overnight at 4 ℃, pouring out the supernatant, drying the precipitate at 60 ℃, and measuring the weight.
And S5, performing 16SrDNA sequence amplification and sequencing on the strain obtained in the step S4, and performing molecular biological identification through Blast and construction of a phylogenetic tree, so as to finally screen out a strain of indigenous beneficial bacteria, wherein the phylogenetic tree is shown in figure 1 and has a closest relationship with paenibacillus.
Through the optical microscope, it was found that: the bacterial cells are straight rod-shaped, are arranged singly, are gram-positive, are uniformly colored and have spores, the bacterial strain is finally identified as the Paenibacillus, is named as C1, is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, and is suggested to be classified and named as the Paenibacillus sp, the preservation number is CGMCC No.20154, and the time is 28 days 6 months 2020.
The invention also aims to apply the multifunctional bacterium C1 to the process of modifying soda saline-alkali soil by using water channel straws, which comprises the following specific steps: the bacterium C1 is sequentially subjected to activation, enrichment and seed tank culture to obtain a bacterium solution containing active bacterium cells, and the bacterium solution is loaded on rice straws mixed with ammonium sulfate and calcium phosphate, wherein the number of the active bacterium cells is 107And (3) mixing the treated water channel straws with 10-20cm of soda saline-alkali soil on the surface layer after the CFU/g is higher than the CFU/g.
As a more excellent technical scheme of the invention, the rice straws are crushed to be less than 5 cm.
As a more preferable technical scheme of the invention, the bacterial liquid usage amount of the bacterium C1 is 100-200 kg/mu.
As a more excellent technical scheme of the invention, the usage amount of the ammonium sulfate is 4-8 kg/mu.
As a more excellent technical scheme of the invention, the usage amount of the calcium phosphate is 4.3-8.6 kg/mu.
As a more preferable technical scheme of the invention, the environment temperature of the mixture of the treated water channel straws and 10-20cm of soda saline-alkali soil on the surface layer is above 20 ℃ and the time is 40-60 days.
As a more preferable technical scheme of the invention, the activation, enrichment and seeding tank culture media are all LB culture media. The LB medium consists of 10g tryptone, 10g NaCl, 5g yeast extract powder and 1L distilled water, and the pH is adjusted to 7.
The beneficial effects are that:
the multifunctional bacterium C1 provided by the invention is an indigenous beneficial bacterium capable of improving soda saline-alkali soil, has strong tolerance to saline-alkali soil, and has the capabilities of producing acid-soluble phosphorus, cellulase and exopolysaccharide. After the microbial inoculum is applied to soda saline-alkali soil together with rice straws, ammonium sulfate and calcium phosphate, the degradation of the rice straws in the soda saline-alkali soil, the formation of soil aggregates with large particle size and water stability and the generation of organic phosphorus in the soil can be promoted, the content of quick-acting phosphorus in the soil is increased, the pH value and the conductivity of the soil are reduced, and the multi-improvement effect on the soda saline-alkali soil is obvious.
The improved soil can provide more active phosphorus for plant growth, increase the phosphorus content in the plant body, promote the growth of plant root systems, and improve the biomass of the overground part, thereby promoting the sustainable utilization of soda saline-alkali soil.
Therefore, the invention solves the problems of strain resource shortage, single improvement effect and poor durability existing in the process of improving soda saline-alkali soil by utilizing rice straws at present.
Drawings
The accompanying drawings, which form a part of the specification, illustrate embodiments of the present invention or technical solutions in the prior art more clearly, and do not limit the present invention.
FIG. 1 is a phylogenetic analysis diagram of bacterium C1 of the present invention;
FIG. 2 is a graph showing the growth of the strain C1 with different NaCl additions,
FIG. 3 is a graph showing the growth of bacterium C1 under different conditions of initial pH;
FIG. 4 is a graph comparing the change in phosphate concentration with and without addition of C1 and C1;
FIG. 5 is a comparison graph of the degradation rate of rice straw to produce cellulase and exopolysaccharide and to rice straw in two addition modes of rice straw and rice straw plus bacterium C1;
FIG. 6 is an SEM image of degraded rice straw;
FIG. 7 SEM picture of degradation of rice straw by bacterium C1.
Detailed Description
In order to make those skilled in the art better understand the technical solution of the present invention, the following examples are used to illustrate the present invention, but not to limit the scope of the present invention.
Example 1 separation and identification of saline-alkali tolerant multifunctional bacteria;
10g of fresh soda saline-alkali soil collected from Changlin county of Songyuan city, Jilin province is added into a triangular flask filled with 90mL of sterile water and glass beads, oscillating at 28 deg.C and 160r/min for 30min, standing for 10min, adding 1mL of soil suspension into 20mL of improved LB liquid culture medium, culturing for 24h, coating on soil broth solid culture medium by gradient dilution method, culturing in 28 deg.C constant temperature incubator for 2d, selecting single colony with obvious appearance difference, streaking and purifying on soil broth solid culture medium, inoculating the purified strain into fresh LB culture medium to obtain fermentation liquid, preparing into bacterial suspension, inoculating into improved NBRIP culture medium according to 2%, performing shake culture at 28 deg.C and 160r/min for 5d, measuring pH value of the fermentation broth every 24h, and collecting the strain with pH value less than 5.5; inoculating acid-producing strains into a rice straw degradation liquid culture medium, performing oscillation culture at 28 ℃ for 7d, collecting the residual straws in the fermentation liquor, drying and weighing, calculating the straw degradation rate, and reserving strains with the rice straw degradation rate of more than 45%; inoculating the strain to the rice straw degradation liquid culture medium for culture, centrifuging the upper layer culture solution at 8000rpm/min for 15min, mixing the supernatant with anhydrous ethanol at a ratio of 1:3, standing at 4 deg.C overnight, pouring out the supernatant, drying the precipitate at 60 deg.C, and measuring weight. Finally, taking the bacterial strain capable of producing acid, degrading rice straws to produce cellulase and extracellular polysaccharide as a target bacterial strain, observing the colony characteristics, gram stain and the like of the target bacterial strain, identifying and determining the strain species by using a 16S rRNA sequence, and carrying out physiological and biochemical reactions.
Through the research, 29 saline-alkali resistant bacteria are obtained from soda saline-alkali soil, wherein 12 acid-producing bacteria, 5 bacteria for degrading straws to produce cellulase and 2 bacteria for producing extracellular polysaccharide are obtained, a strain with strong acid production capacity, strong rice straw degradation capacity and strong extracellular polysaccharide production capacity is finally screened, the pH of a fermentation liquor of the strain is reduced to 4.60 after the strain is cultured in an improved NBRIP culture medium for 5 days, the phosphate radical concentration is 141.70mg/L, the degradation rate of the rice straws is 54.24% after the strain is cultured in a rice straw degradation liquid culture medium for 7 days, the cellulase activity is as high as 1085.71U/mL, and the degraded rice straws can produce 287.41mg/L of extracellular polysaccharide. After the strain C1 is cultured on a soil broth solid medium at 28 ℃ for 48 hours, a light-yellow, round, opaque, smooth-shaped, viscous and tasteless colony with a slightly convex surface and regular edges can be seen. Physiological and biochemical reactions show that the strain C1 can metabolize D-glucose, D-ribose, D-galactose and the like to produce acid and can react with beta-glucosidase, beta-xylosidase, beta-mannosidase and the like. Phylogenetic trees the phylogenetic relationship between the strains and paenibacillus was closest as shown in figure 1. The optical microscope shows that the somatic cells are in straight rod shape, are arranged singly, are gram-positive, are uniformly colored and have spores.
The result is combined, and the bacterium C1 is finally identified as Paenibacillus sp which is preserved in the China general microbiological culture Collection center
EXAMPLE 2 saline tolerance of Strain C1
Inoculating 1 loop of the strain C1 preserved on a slant culture medium in a refrigerator at 4 ℃ to 5mL of LB culture medium, oscillating at 28 ℃ and 160r/min for 24h, completely inoculating the culture to 100mL of fresh LB culture medium, culturing for 24h under the same conditions to obtain a fermentation liquid, centrifuging the fermentation liquid at 4 ℃ and 8000r/min for 10min, collecting bacterial cells, suspending the bacterial cells in sterile water, and preparing the bacterial cells with the number of viable bacteria not less than 109Inoculating the bacterial suspension of CFU/mL into LB culture medium at a ratio of 2%, changing the addition amount of NaCl to 3%, 5% and 7% by using LB culture medium with pH value of 7, performing shake culture at 28 deg.C and 160r/min for 4 days while preparing LB culture medium without NaCl addition and under normal conditions (NaCl addition amount is 1%), and measuring OD of the fermentation liquid with 722-visible spectrophotometer600(ii) a Adjusting the pH of LB medium to 8, 9, 10, and 11 respectively with 1mol/L NaOH with NaCl addition of 1%, shake-culturing at 28 deg.C and 160r/min for 5d, and measuring OD of the fermentation broth with 722-visible spectrophotometer600。
The growth conditions of the strain C1 under the conditions of different NaCl addition amounts are shown in figure 2, and it can be seen from the figure that the strain C1 has higher salt tolerance, and when the NaCl addition amount is 1%, OD is6000.891, OD when NaCl addition amount reached 7%600At 0.557, growth was still maintained. The growth of strain C1 under different conditions of initial pH is shown in FIG. 3, from which it can be seen that the OD is at 11 when the initial pH of the medium is600At 0.255, strain C1 still maintained good growth. The experiments show that the strain C1 has good saline-alkali tolerance.
Example 3 Strain C1 ability to produce acid-soluble phosphorus, degrade rice straw and produce exopolysaccharides
Inoculating 1 loop of the strain C1 preserved on a slant culture medium in a refrigerator at 4 ℃ to 5mL of LB culture medium, oscillating at 28 ℃ and 160r/min for 24h, completely inoculating the culture to 100mL of fresh LB culture medium, culturing for 24h under the same conditions to obtain a fermentation liquid, centrifuging the fermentation liquid at 4 ℃ and 8000r/min for 10min, collecting bacterial cells, suspending the bacterial cells in sterile water, and preparing the bacterial cells with the number of viable bacteria not less than 109The bacterial suspension of CFU/mL is inoculated into an improved NBRIP culture medium (strain C1) according to the proportion of 2 percent, meanwhile, the control treatment (CK) without adding bacteria is carried out, the oscillating culture is carried out for 5 days under the conditions of 28 ℃ and 160r/min, samples are taken every 24 hours, and the pH value and the phosphate radical content of the fermentation liquor of the control and the strain C1 are determined. Inoculating a bacterial suspension prepared from a fermentation liquor obtained by activating and seed culturing the strain C1 into a rice straw degradation liquid culture medium, performing control treatment without adding bacteria, culturing for 7d at 28 ℃ and 160r/min, sampling every 24h, performing autoclaving at 121 ℃ for 15min by taking 10mL of strain C1 fermentation liquor as a control (dead bacteria) for determining the activity of the CMC enzyme, cooling to room temperature, centrifuging the inactivated bacteria fermentation liquor and the inactivated bacteria fermentation liquor at 8000r/min for 10min, respectively taking 0.5mL of the inactivated bacteria fermentation liquor and placing the inactivated bacteria fermentation liquor into a 50mL colorimetric tube, and determining the activity of the cellulase by using a DNS method; centrifuging the rest fermentation liquid of control and strain C1 at 8000r/min for 15min, mixing the supernatant with anhydrous ethanol at a ratio of 1:3, standing at 4 deg.C overnight, taking out, pouring out the supernatant, repeatedly washing the precipitate with anhydrous ethanol for 2-3 times, dissolving with small amount of distilled water, collecting the filtrate1mL of the liquid is put into a 25mL colorimetric tube, and the content of the extracellular polysaccharide is measured by a phenol-sulfuric acid method. Inoculating a bacterial suspension prepared from fermentation liquor obtained by activating and seed culturing the strain C1 into a rice straw degradation liquid culture medium, performing control treatment without adding bacteria, degrading for 7d at 28 ℃ and 160r/min, pouring out liquid in the culture medium, taking out the rice straws, washing with sterile water for 2-3 times, placing in an oven, drying to constant weight, weighing, calculating the removal rate, and observing the dried raw rice straws and the rice straws degraded by the strain C1 by a Scanning Electron Microscope (SEM).
The phosphorus dissolving capacity of the strain C1 is shown in FIG. 4, and after the strain C1 is cultured in the improved NBRIP culture medium for 5d, the strain C1 can reduce the pH of the fermentation liquor from 7.0 to 4.60, and the phosphate concentration is 141.70mg/L, while the pH and the phosphate concentration of a control are unchanged. The degradation rate of the strain C1 for degrading rice straws to produce cellulase (CMC enzyme) and extracellular polysaccharide and degrading rice straws is shown in figure 5, after the strain C1 is cultured in a rice straw degradation liquid culture medium for 7 days, the activity of the CMC enzyme is as high as 1085.71U/mL, the content of the extracellular polysaccharide is 287.41mg/L, and the removal rate of the rice straws is 54.24%.
The original rice straw SEM image in the rice straw liquid culture medium is as shown in figure 4, and it can be seen that the wax layer of the surface structure of the original rice straw is smooth and regular, the straw structure is relatively complete and closely connected, and the fiber bundle is uniformly extended; the structure of the rice straw degraded by the strain C1 is changed, the flat surface is disturbed, the compact structure becomes loose, the rice straw is decomposed into a regular block structure, and the original structure of the straw is damaged.
Example 4 Strain C1 synergistic improvement of Rice straw on soda saline-alkali soil
Fresh soda saline-alkali soil is collected from an Anzhen town of Songyuan city, Jilin province, large stones, plant residues and other impurities are removed, the soil is naturally dried, and a soil culture test is carried out after the soil is sieved by a 2mm sieve. Inoculating a strain C1 preserved on a slant culture medium in a refrigerator at 4 ℃ to an LB culture medium, performing activated culture for 24h under the conditions of 28 ℃ and 160r/min, inoculating the strain C1 to a fresh LB culture medium according to the proportion of 2%, culturing for 24h, centrifuging for 10min at 4 ℃ and 8000r/min, cleaning somatic cells with sterile water for 2-3 times, then resuspending the somatic cells in the sterile water with a proper amount of sterile water to prepare a bacterial suspension, spraying 750mL of the bacterial suspension on the surface of 150g of rice straws mixed with 6g of ammonium sulfate and 6.5g of calcium phosphate, then adding the mixture to soil, uniformly mixing, simultaneously performing control treatment (CK) without adding bacteria, replacing the bacterial suspension with the sterile water, performing the same treatment process, and repeating each treatment for 3 times, wherein the specification of a plastic basin of a soil culture container is as follows: the method comprises the steps of culturing at 25 ℃, periodically adding sterile water to maintain the water content of soil to be about 40%, turning the soil 1 time every 15 days, collecting a soil sample after culturing for 50 days, naturally drying, removing undecomposed straws in the soil from 2kg of air-dried soil by using an electrostatic adsorption method, repeatedly washing the undecomposed straws by using the sterile water, drying and weighing, calculating the degradation rate of the straws, and then measuring the composition of soil aggregates, the stability of the aggregates in water, the pH value and the conductivity of the soil, the content of organic carbon, the content of available phosphorus and the content of organic phosphorus in the soil of each grain size fraction by using the air-dried soil from which the straws are removed.
The result shows that after the bacterial strain C1, the rice straws, the ammonium sulfate and the calcium phosphate are added into the soda saline-alkali soil, compared with CK, the bacterial strain C1 can better play a role in the saline-alkali soil, and the degradation rate of the rice straws in the soil reaches 51.16%. The pH value of each size fraction aggregate is reduced to 8.24 at the lowest, the conductivity is reduced to 0.225mS/cm at the lowest, the stability of the aggregate in water is further enhanced, and the weight loss rates of the size fraction aggregates larger than 0.25mm and 0.25-0.053mm are respectively reduced by 2.65 percent and 3.58 percent compared with CK, so that the obvious level is achieved; the formation of large aggregates is promoted, and the content of the aggregates in the soil with the grain size of more than 0.25mm is obviously increased by 4.13 percent; the content of the soil aggregate with the grain size of 0.25-0.053mm and less than 0.053mm is respectively and obviously reduced by 3.25 percent and 1.88 percent; meanwhile, the total organic carbon content of the aggregate with the grain size of more than 0.25mm, 0.25-0.053mm and less than 0.053mm is respectively increased by 35.69 percent, 36.78 percent and 25.44 percent; the water-soluble organic carbon content of the aggregate with the grain size of more than 0.25mm and 0.25-0.053mm is respectively and obviously increased by 6.14 percent and 3.58 percent; increasing the content of the available phosphorus and the organophosphorus in the aggregate of each grade, wherein the content of the available phosphorus is increased within the range of 29.49-38.72 percent, and the content of the organophosphorus is increased within the range of 39.26-44.97 percent. The experimental results show that the strain C1, the rice straws, the ammonium sulfate and the calcium phosphate are added into the soda saline-alkali soil together, the physicochemical property of the soda saline-alkali soil is greatly improved, the strain has an important application value in the aspect of improving the soda saline-alkali soil by using microorganisms, and theoretical basis and technical support are provided for the compound improvement of the soda saline-alkali soil by using the paenibacillus C1, the rice straws, the ammonium sulfate and the calcium phosphate.
Example 5 Effect of soda saline-alkali soil modification on growth of oilseed rape
Performing field test in 5 months in 2020 and 5m near Anzhen town of Songyuan city of Jilin province, wherein the area of a cell is 5m multiplied by 6m, crushing rice straws into small sections with the size of below 5cm, paving the small sections on the surface of saline-alkali soil according to the using amount of 150 kg/mu, respectively spreading ammonium sulfate and calcium phosphate according to the using amounts of 6 kg/mu and 6.4 kg/mu in sequence, and spraying the prepared bacterial strain C1 bacterial suspension on the surface of the rice straws to ensure that the number of viable bacteria per g of straws is 1 multiplied by 107CFU/g, then mixing with 0-20cm surface soil uniformly, performing contrast treatment without adding the strain C1, repeating each treatment for 3 times, adding water during improvement to keep the soil moist, planting rape 45d later according to a conventional method, measuring the germination rate, growing again 45d later after thinning, harvesting, selecting 20 rapes with basically consistent growth potential in each cell by using a diagonal method during harvesting, and measuring the biomass of the overground part, the maximum root length, the phosphorus content of the plants and the chlorophyll content in the leaves.
Results show that compared with control treatment, the germination rate of the rape is increased by 16.25%, the longest root length is increased by 17.64% on average, leaf chlorophyll is increased by 8.9 SPAD values, the dry weight of the overground part is increased by 26.87%, the phosphorus content of the plant is increased by 8.5%, the bacteria C1, in cooperation with the rice straw, improve the soda saline-alkali soil, obviously improve the germination rate of the rape and promote the growth of the rape, and the improvement effect is very obvious.
Claims (8)
1. A multifunctional bacterium C1 for improving soda saline-alkali soil, which is characterized in that: the strain C1 has been deposited in China general microbiological culture Collection center (CGMCC) at 28 th 6 th 2020, the preservation number is CGMCC No.20154, and the proposed classification is named as Paenibacillus sp.
2. The use of the multifunctional bacterium C1 for improving soda saline-alkali soil as claimed in claim 1 in the field returning of rice straw to field.
3. The application of claim 2, comprising the following steps:
(1) crushing rice straw, spreading on the surface of soda saline-alkali soil,
(2) uniformly spraying ammonium sulfate and calcium phosphate on the surface of the flatly laid rice straws;
(3) uniformly spraying a bacterial liquid, turning into surface soda saline-alkali soil, and uniformly mixing, wherein the bacterial liquid is obtained by activating and enriching culture of a bacterium C1;
(4) and (5) planting plants after decomposing the rice straws.
4. Use according to claim 2, characterized in that: the bacterial liquid usage amount of the bacteria C1 is 100-200 kg/mu.
5. The use of claim 2, wherein the amount of ammonium sulfate used is 4-8 kg/acre.
6. The use according to claim 2, wherein the amount of calcium phosphate used is 4.3-8.6 kg/acre.
7. The use of claim 2, wherein said rice straw is crushed to less than 5 cm.
8. The use of claim 2, wherein the rice straw decomposition temperature is above 20 ℃ and the time is 40-60 days.
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