CN118086138A - Burkholderia cepacia and application thereof in reducing cadmium and promoting growth of rice - Google Patents
Burkholderia cepacia and application thereof in reducing cadmium and promoting growth of rice Download PDFInfo
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- 229910052793 cadmium Inorganic materials 0.000 title claims abstract description 74
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 title claims abstract description 74
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Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the technical field of biology, and particularly relates to burkholderia cepacia and application thereof. The invention discloses Burkholderia cepacia (Burkholderia cepacia) WW13, which has the preservation number: CGMCC No.27717. The invention also discloses application of the Burkholderia cepacia (Burkholderia cepacia) WW13 in promoting safe production of rice under cadmium stress.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to burkholderia cepacia and application thereof.
Background
As human activities such as mining, agriculture, and sewage treatment are increased, global farmland soil is severely affected by heavy metal pollution. Cadmium is one of the heavy metals causing farmland soil pollution, and has serious biotoxicity. Cadmium easily migrates into the soil and is then absorbed by plants, which can lead to a number of physiological and biochemical problems. In addition, cadmium in crops can be transmitted to humans through the food chain and cause health problems such as chronic damage to the kidneys and bones. Thus, controlling soil cadmium pollution and reducing cadmium uptake into crop edible tissues are significant challenges facing environmental science. According to previous researches, methods for reducing cadmium in soil can be classified into physical and chemical restoration techniques and biological restoration techniques, but physical and chemical restoration methods have high cost and usually cause secondary pollution. Phytoremediation methods in bioremediation technology, such as plant stabilization, plant volatilization and plant extraction, are safe and reliable. However, this approach is not practical for social and agricultural needs because it is slow and phytoremediation methods generally require long periods of non-use of cultivated land. There is therefore a need to develop techniques that can directly reduce cadmium accumulation in the edible parts of crops without interrupting soil use.
Nitrogen is an important nutrient element for the growth and development of rice, influences the tillering and spike quality of the rice, further influences the yield of the rice, and has important significance for high and stable yield of the rice. Based on previous studies, it was found that nitrogen fertilizer can reduce cadmium accumulation in crops by accelerating crop growth. In particular, the use of ammonium fertilizers helps to inhibit cadmium uptake, xylem transport and subsequent accumulation in rice. However, in rice production in China, too much nitrogen fertilizer is put into the production to ensure the yield, so that the nitrogen fertilizer utilization rate is reduced, and eutrophication and water pollution are easily caused.
Microorganisms are increasingly valued in recent years in terms of relieving the toxicity of heavy metals to crops and improving the stress resistance of crops to heavy metal stress due to the characteristics of multiple functions, strong adaptability, no secondary pollution, symbiosis with crop roots and the like. Some rhizosphere growth promoting bacteria with excellent adaptability and good repairing effect can secrete substances such as siderophores, change the bioavailability of heavy metals, increase the absorption and removal rate of the heavy metals, and are beneficial to reducing the toxic effect of the heavy metals on plants. Azotobacter is used as a common growth-promoting bacterium in farmlands, and is important for absorbing nitrogen elements of crops in the farmlands. The microorganisms can also provide additional nitrogen nutrient elements by symbiotic microorganisms, can accelerate crop growth by reducing the available time for heavy metal accumulation, and can reduce the harm of heavy metal pollution and not easily cause secondary pollution. And the related researches on the microbial restoration of the azotobacter for promoting the growth of rice and reducing the cadmium absorption are less.
The invention of CN116836895A, a heavy metal contaminated soil remediation microbial inoculum, a preparation method and application thereof, informs: a heavy metal contaminated soil remediation microbial inoculum comprises pseudomonas putida AM21 and Burkholderia (Burkholderia) AM20, wherein the ratio of the viable count of the two is 1 (0.2-5), and the total viable count is (1-5) multiplied by 109cfu/g. The screened Burkholderia AM20 can activate heavy metals in soil and promote the plants to absorb the heavy metals in the soil; the screened pseudomonas putida AM21 has the effect of phosphate dissolution, can improve the growth quantity of plants and promote the absorption of the plants to heavy metal cadmium. The heavy metal contaminated soil remediation microbial inoculum combines the functions of activating soil heavy metal by Burkholderia AM20 and promoting heavy metal absorption by Pseudomonas putida AM21, and has outstanding remediation effect on cadmium contaminated soil without continuous input of exogenous heavy metal.
The comparative example 3 microbial inoculum only added with Burkholderia AM20 has higher enrichment coefficient for cadmium, the comparative example 4 microbial inoculum only added with Pseudomonas putida AM21 has lower enrichment coefficient for cadmium, but the growth amount of plants is more obviously increased, which indicates that the two strains have the functions of promoting the enrichment of the plants for cadmium and promoting the growth of the plants when being singly used, the Burkholderia AM20 can activate heavy metals in soil, promote the absorption and enrichment of the plants for the heavy metals, and the plant growth promoting effect of the Pseudomonas putida AM21 is more obvious, thereby relieving the cadmium stress, promoting the growth of the plants and finally improving the removal rate of the cadmium.
CN112662588a, a conifer rhizosphere biocontrol growth-promoting bacterium, two-way burkholderia strain and application thereof, informs: the conifer rhizosphere biocontrol growth-promoting bacterium, namely the bidirectional burkholderia (Burkholderiaambifaria) strain ZB-155, has IAA production and phosphate dissolving capabilities and has remarkable promotion effect on the growth of conifer such as pinus sylvestris; the strain also has the capability of secreting cellulase and protease, and the strain and a fermentation product thereof have antibacterial effects on pathogenic bacteria such as seedling damping-off, poplar rot, pine tip disease, hazelnut leaf damping-off, hazelnut brown spot and the like, and can improve plant rhizosphere soil nutrient components, so the invention provides application of the strain and a derivative product thereof in plant planting and soil improvement.
The above Burkholderia sp.AM 20 was used in a soil with a concentration of 3.32mg/kg, which was not informed of its tolerance to high concentration cadmium, and which was used for activation of cadmium in the soil. Burkholderia (Burkholderiaambifaria) strain ZB-155 is only concerned with the inhibition of pathogenic bacteria and is not known to have high levels of cadmium tolerance. Therefore, all the 2 strains are not suitable for safe production and utilization of rice under cadmium stress.
Disclosure of Invention
The invention aims to provide a strain-Burkholderia cepacia (Burkholderia cepacia) WW13 for promoting rice growth and reducing cadmium enrichment under cadmium stress and application thereof.
In order to solve the technical problems, the invention provides Burkholderia cepacia (Burkholderia cepacia) WW13 with the preservation number: CGMCC No.27717.
The 16S rDNA gene sequence of the Burkholderia cepacia (Burkholderia cepacia) WW13 is shown as SEQ ID NO. 1.
The invention also provides application of the Burkholderia cepacia (Burkholderia cepacia) WW 13: high tolerance and cadmium adsorption capacity, has the function of nitrogen fixation, can promote rice growth and nitrogen absorption, and reduces rice grain cadmium accumulation.
In summary, the invention provides an application of Burkholderia cepacia (Burkholderia cepacia) WW13 isolated from farmland soil polluted by the periphery of a waste dismantling site in promoting safe production of rice under cadmium stress.
The Burkholderia cepacia WW13 is separated from cadmium-polluted farmland soil around a Taizhou waste electric appliance dismantling site in Zhejiang province, and the preservation information is as follows:
preservation name: burkholderia cepacia Burkholderia cepacia, accession number: china general microbiological culture Collection center, preservation address: beijing, chaoyang district North Star, west road 1, 3, accession number: CGMCC No.27717, the preservation time is 2023 and 27 days of 06 months.
The colony characteristics of this strain were as follows: the bacterial strain is bacillus, has a rough surface, no flagellum and gram negative, has a thallus size of (0.9-1.6) mu m x (0.2-0.5) mu m, and has a circular colony, a convex and smooth surface, a complete edge, a yellow color and opaqueness on a solid culture medium.
The strain is obtained by screening collected soil surrounding a disassembly field of a waste electric appliance in Taizhou city of Zhejiang province, and is proved to be Burkholderia cepacia according to a 16S rDNA sequencing identification method.
The invention also relates to application of the Burkholderia cepacia (Burkholderia cepacia) WW13 in cadmium pollution. Specifically, the strain is used for adsorbing free cadmium, promoting rice growth and nitrogen accumulation, and reducing cadmium accumulation in seeds. The strain is inoculated into a culture medium containing cadmium ions, and is cultured for 3 days under the aerobic condition of 28 ℃ and 180r/min, so that the cadmium ions can be effectively removed. The rice is inoculated to the root system of the rice in the water planting system, so that the growth of the rice can be effectively promoted, the nitrogen content in plants is improved, and the cadmium accumulation in seeds is reduced.
The cadmium-containing culture medium mainly refers to an LB liquid culture medium mainly containing free cadmium ions, and the cadmium concentration is 0-1000mg/L. In the water culture system, the cadmium concentration is 0, 0.3 and 2.0mg/L respectively.
The strain can be used for adsorbing cadmium ions, and is cultured for 3 days under the conditions of aerobic conditions, pH of 7.0, temperature of 25-30 ℃, OD 600 1.0.0, cadmium concentration of 100mg/L and rotating speed of 180r/min, centrifugal filtration is carried out, the removal rate of the cadmium ions is 75%, and the concentration of the cadmium ions is reduced to 25.35mg/L. Under the water culture condition, the dry weight and the nitrogen content of the rice are obviously increased, wherein the cadmium content of the seeds in the 0.3mg/L treatment is reduced from 3.78mg/kg to 2.90mg/kg, and the cadmium content of the seeds in the 2.0mg/L treatment is reduced from 16.00mg/kg to 10.90mg/kg.
The invention designs a strain of high-tolerance cadmium-resistant and nitrogen-fixing burkholderia cepacia (Burkholderia cepacia) WW13 separated from farmland soil polluted by the periphery of a waste electric appliance dismantling field and application of the strain in promoting safe production of agricultural products by cadmium stress, which belongs to the first discovery.
In conclusion, the invention obtains the Burkholderia cepacia (Burkholderia cepacia) WW13 for rice growth under cadmium stress and reducing cadmium accumulation through screening, has the functions of high tolerance to cadmium and nitrogen fixation, can promote rice growth, promote rice nitrogen absorption, reduce the potential of cadmium accumulation in seeds, and provides a technical basis for ensuring safe production and utilization of rice in cadmium polluted farmlands and increasing yield, and has wide application prospect.
Drawings
The following describes the embodiments of the present invention in further detail with reference to the accompanying drawings.
FIG. 1 shows Burkholderia cepacia (Burkholderia cepacia) WW13 (magnification 20000 times);
FIG. 2 is a 16S rDNA-based phylogenetic tree of strains;
FIG. 3 shows growth curves for Burkholderia cepacia (Burkholderia cepacia) WW13 at various cadmium concentrations;
FIG. 4 shows the effect of Burkholderia cepacia (Burkholderia cepacia) WW13 on dry weight of rice in a hydroponic system;
FIG. 5 shows the effect of Burkholderia cepacia (Burkholderia cepacia) WW13 on nitrogen content in rice in a hydroponic system;
FIG. 6 is the effect of Burkholderia cepacia (Burkholderia cepacia) WW13 on rice grain cadmium accumulation in a hydroponic system.
Detailed Description
The invention will be further described with reference to the following specific examples, but the scope of the invention is not limited thereto:
example 1 screening and identification of tolerant strains
1. Materials and methods
1.1 Culture medium and reagent
LB medium: yeast extract 5g/L, tryptone 10g/L, naCl 10g/L, pH=7, 121 ℃, 15min sterilization.
CAS medium: each 100mL of the solution contains 1mL of 20% sucrose solution, 3mL of 10% acid hydrolyzed casein, 1mmol/L CaCl 2100μL,1mmol/L MgSO4·7H2 O2 mL, 1.8g of agar and the balance deionized water. The above% is mass%.
Phosphate buffer (pH 6.5) and CAS dye were slowly added 5mL each at about 60℃when the CAS medium had not yet completely coagulated, and after cooling to room temperature, a blue solid detection medium was obtained.
All solutions were formulated with deionized water.
CAS solution: 0.7g of CAS (chromium azure) is measured and dissolved in deionized water, the volume is fixed to 500mL, and the solution is stored in a reagent bottle for standby.
FeCl 3 solution: 0.27g FeCl 3 0.83.83 mL of concentrated HCl (12 mol/L) is taken and dissolved in a proper amount of deionized water, the volume is fixed to 1000mL, and the mixture is stored in a reagent bottle for standby.
HDTMA solution: 1.5g HDTMA g was dissolved in a suitable amount of deionized water and the volume was set to 1000mL.
CAS dye liquor: adding 100mL of FeCl 3 into 500mL of CAS solution, shaking uniformly to obtain solution A, adding 400: 400mL HDTMA solution into HDTMA solution slowly along the wall of a beaker, and mixing the two solutions uniformly by gentle shaking to obtain the CAS dye liquor.
All media in the present invention need to be routinely sterilized prior to use, namely: sterilizing at 115 deg.c for 15 min.
1.2 Selection of tolerant strains
1.2.1 Strain Source
Collecting cadmium-polluted farmland soil around waste electric appliance dismantling site in Zhejiang province Taizhou
1.2.2 Isolation, purification and screening of resistant strains
2G of each cadmium-polluted farmland soil was taken out and placed into 50mL triangular flasks of 18mL of sterile water under sterile conditions. Shaking on a 180r/min shaker at 28deg.C for 1 hr, serial dilution in sterile water, placing 100 μl volume of diluted soil suspension on blue solid detection medium, and culturing at 28deg.C in incubator for 7 days. Single bacterial colony with large orange halo is picked up, repeated plate streak separation and purification are carried out until screening is carried out to obtain single bacterial colony, the single bacterial colony is inoculated into LB liquid culture medium, shake culture is carried out in a shaking table until turbidity is achieved, 900 mu L of bacterial liquid is taken according to the ratio of 1:1 are added into a preservation tube containing 50 percent of glycerol and are put into a refrigerator with the temperature of minus 80 ℃ for preservation.
One of the selected strains was designated WW13.
1.2.3WW13 identification of Strain
The strain identification was verified using 16S rDNA sequencing identification method. The total DNA of WW13 strain is used as template, the general primer of 16SrDNA gene is used for PCR amplification, the obtained amplified fragment is recovered and sequenced to determine the size of the amplified fragment, and then the sequencing result is subjected to homology comparison with the sequence in NCBI by using MEGA software.
The strain WW13 has high homology with Burkholderia cepacia (Burkholderia cepacia), the homology is close to 99%, and the genetic distance is close (figure 2).
Morphological feature observation and physiological and biochemical characteristic measurement of 1.2.4WW13 strain
Inoculating the strain in an LB liquid culture medium, performing pretreatment for 72 hours, and observing morphological characteristics of the strain by adopting a scanning electron microscope; gram staining was performed on the purified strain during the log phase of growth.
The bacteria are bacillus, the surface is rough, no cytoblast is produced, no flagellum exists, gram negative exists, the bacterial size is about long (0.9-1.6) mu m, wide (0.2-0.5) mu m (as shown in figure 1), and the bacterial colony is round, convex and smooth on the surface, complete in edge, yellow and opaque on a solid culture medium.
1.3 Determination of the function of the WW13 Strain
Determination of cadmium tolerance and adsorption Capacity of 1.3.1WW13 Strain
Inoculating (transferring) 0.2mL of WW13 strain stored in glycerol at-80 ℃ into 20mL of sterilized LB liquid medium, pre-culturing for 48 hours by constant temperature gas bath oscillation (28 ℃,180 r/min), transferring into a sterilized centrifuge tube, centrifuging at low temperature (4 ℃,6000g/min,5 min), collecting thalli, taking LB liquid medium as a blank, diluting the bacterial liquid to OD 600 =1, taking 1mL of LB liquid medium with different Cd concentrations (the concentration is shown in figure 3), culturing, and determining the value of OD 600 in 0, 6,12, 24, 36, 48, 60 and 72 hours by experiment setting 3 times.
The cadmium tolerance concentration of the Burkholderia cepacia WW13 obtained by screening is shown in a figure 3, and the maximum cadmium tolerance concentration can reach 1000mg/L. The remaining strains obtained in the screening process of the present invention and the currently existing Burkholderia (as shown in Table 1 below) were subjected to the cadmium resistance test according to the above method after replacing WW13, and the results obtained are compared with those of WW13 as shown in Table 1 below.
TABLE 1 comparison of cadmium tolerance concentration of Burkholderia cepacia WW13 with other strains
Description: the WW14, WW12, WW3 and WW23 obtained in the screening process are Burkholderia territorii, burkholderia metallica, pseudomonas kribbensis and Bacillus aequororis respectively.
To test the cadmium adsorption capacity of the strain, 1mL of the bacterial liquid was added to 100mL of LB medium containing 100mg/L of Cd in accordance with the aforementioned activation and transfer steps. The culture condition is 28 ℃ and 180r/min; after 72 hours, samples were collected and centrifuged (12,000 g/min,5 min) to separate supernatant and particles. The supernatant was filtered through a 0.22 μm filter and the Cd concentration of the supernatant was measured by ICP-MS. The ICP-MS assay was a reference HJ 776-2015.
Culturing for 3 days under aerobic condition at pH 7.0, temperature of 28deg.C, OD 600 1.0.0, cadmium concentration of 100mg/L and LB liquid culture condition with rotation speed of 180r/min, centrifuging, filtering to remove cadmium ion at 75%, and reducing cadmium ion concentration of supernatant to 25.35mg/L. The separated and screened Burkholderia cepacia WW13 can effectively adsorb free cadmium ions, and has certain potential in the field of relieving the cadmium stress of rice.
1.3.2 Determination of Nitrogen fixation Capacity of Strain
The content of the azotase is determined by using a kit (a Kett kit). 1) Extracting: inoculating (transferring) 0.2mL of WW13 strain stored in glycerol at-80 ℃ into 20mL of sterilized LB liquid medium, pre-culturing for 48 hours by constant temperature gas bath oscillation (28 ℃ C., 180 r/min), transferring into a sterilized centrifuge tube, centrifuging at low temperature (4 ℃ C., 12000g/min,5 min) to collect thalli, diluting the thalli to OD 600 =1 by taking PBS buffer as a blank, and adding the thalli into the Abbe's liquid medium according to a proportion of 1% for culturing for 72 hours under the condition of 28 ℃ C., 180r/min. Bacterial culture in the Abbe's liquid culture medium was centrifuged at 4℃for about 20 minutes, and the supernatant was collected. 2) Sample adding: blank holes (blank control holes are not added with samples and enzyme-labeled reagents, and the rest steps are the same), standard holes and sample holes to be tested are respectively arranged. And (3) accurately adding 50 mu L of standard substances on the enzyme-labeled coated plate, adding 40 mu L of sample diluent into a sample hole to be detected, and then adding 10 mu L of sample to be detected (the final dilution of the sample is 5 times). 3) Incubation: the plates were then covered with a plate membrane and incubated at 37℃for 30 minutes. 4) Preparing liquid: the 20-fold concentrated washing solution is diluted with distilled water for later use. 5) Washing: carefully removing the sealing plate film, discarding the liquid, spin-drying, filling each hole with the washing liquid, standing for 30 seconds, discarding, repeating the process for 5 times, and beating. 6) Adding enzyme: 50. Mu.L of enzyme-labeled reagent was added to each well, except for blank wells. 7) Re-incubation, washing, procedure 3) and 5); 8) Color development: adding 50 mu L of a color developing agent A and 50 mu L of a color developing agent B into each hole, gently shaking and uniformly mixing, and developing for 10 minutes at 37 ℃ in a dark place; 9) And (3) terminating: the reaction was stopped by adding 50. Mu.L of stop solution to each well (blue turned yellow immediately). 10 Determination: the absorbance (OD value) of each well was measured sequentially at the wavelength of blank Kong Diaoling, 450 nm. The measurement should be performed within 15 minutes after the addition of the stop solution.
Drawing a standard curve by taking the concentration of the standard substance as an abscissa and the OD value as an ordinate, and finding out the corresponding concentration from the standard curve according to the OD value of the sample; multiplying by the dilution factor; or calculating a linear regression equation of the standard curve by using the concentration and the OD value of the standard substance, substituting the OD value of the sample into the equation, and calculating the concentration of the sample.
The standard curve of the activity of the azotase is as follows: c= 10828.3669 ×abs+5.2594, r 2 = 0.9996, the measured data and standard curve data are shown in tables 2 and 3, and the colony enzyme activity concentration of the strain is 324.077 (IU/L).
TABLE 2 Burkholderia cepacia WW13 Nitrogen fixation enzyme Activity
TABLE 3 Standard curve
| Standard curve | |
| Abs | Concentration IU/L |
| 0.0006 | 10 |
| 0.0012 | 20 |
| 0.0032 | 40 |
| 0.0069 | 80 |
| 0.0143 | 160 |
In conclusion, 1 strain of bacteria with high tolerance to cadmium is obtained through separation, purification and screening, and is named as WW13, the maximum tolerance concentration of the bacteria is 1000mg/L, and the bacteria are aerobically cultured for 3 days, so that the removal rate of the bacteria to 100mg/L is found to be 75%.
Comprehensive physiological and biochemical properties and the evolution analysis of the 16S rDNA system can show that the strain belongs to Burkholderia cepacia (Burkholderia cepacia).
Namely, 1 strain of high-efficiency strain (CGMCC No. 27717) which can endure and adsorb cadmium and has nitrogen fixation capability is separated from cadmium-polluted farmland soil around a Taizhou waste electric appliance dismantling site in Zhejiang province, and is identified as Burkholderia cepacia (Burkholderia cepacia) through 16S rDNA.
The information on the preservation of WW13 is as follows:
preservation name: burkholderia cepacia Burkholderia cepacia, accession number: china general microbiological culture Collection center, preservation address: beijing, chaoyang district North Star, west road 1, 3, accession number: CGMCC No.27717, the preservation time is 2023 and 27 days of 06 months.
Example 2 verification of Strain to promote Rice growth and reduce grain cadmium accumulation
1. Materials and methods
1.1 Culture medium and reagent
LB liquid medium: 5g/L yeast extract, 10g/L peptone, 10g/L NaCl, pH 7.0, the balance deionized water.
The rice water planting nutrient solution comprises: according to the formula configuration of the conventional nutrient solution of the International Rice institute (IRRI), 800 times of the mother liquor components (g/L) are as follows:
mother liquor I: NH 4NO3 91.6、CaCl2 88.6.6;
Mother liquor II: naH 2PO4·2H2O 40.3、K2SO4 71.4.4;
Mother liquor III: mgSO 4·7H2 O324;
Mother liquor Ⅳ：MnCl2·4H2O 1.5、H3BO3 0.934、CuSO4·5H2O 0.031、(NH4)6·Mo7O24·2H2O 0.074( ammonium heptamolybdate), znSO 4·7H2O 0.035、FeCl3·6H2 O7.7, citric acid monohydrate C6H8O7.H 2 O11.9 and concentrated sulfuric acid 50mL.
Every 2L of rice culture solution is respectively added with 5mL, 5mL and 4mL of mother liquor I, mother liquor II, mother liquor III, mother liquor IV and the balance deionized water, and the pH is adjusted to be 4.5-5.0.
Cadmium treatment: cadmium mother liquor with concentration of 10000mg/L is respectively added into each 2L of rice culture solution, so that the water culture conditions are respectively 0.3 and 2.0mg/L of cadmium stress treatment (namely, the cadmium mother liquor is used as nutrient solution of 0.3 and 2.0mg/L Cd (NO 3)2).
1.2 Effect of the Strain on Rice growth and grain cadmium content
The activated WW13 bacteria are picked up and inoculated into a 250mL triangular flask filled with 100mL LB liquid culture medium, and placed in a shaking table (28 ℃ C., 180 rmp/min) for shaking culture for 36h until the OD value of the bacterial liquid is between 1.0 and 1.5; centrifuging the bacterial liquid (12,000 g/min,5 min), pouring out the supernatant, and collecting the bacterial cells; the collected thalli are washed by a resuspension centrifugation method, washed 3 times by using sterile water, and then the OD value of WW13 bacterial liquid is adjusted to 1.0 by using the sterile water for later experiment.
Transferring three uniform and healthy rice seedlings into 0.3 and 2.0mg/L Cd (NO 3)2) nutrient solution, inoculating the bacterial solution into a rice root system to ensure that the concentration of the bacterial solution in a water culture basin is 10 7 CFU/mL, changing the nutrient solution every week, continuously culturing for 104 days by adding the bacterial solution according to the same steps, harvesting plant samples, deactivating enzymes at 105 ℃ for half an hour, drying at 75 ℃ to constant weight, weighing and crushing each part of the plant, measuring the nitrogen content of each part by an elemental analyzer, and measuring the cadmium content of rice grains by a microwave sterilization method.
2 Results
The experimental results are shown in fig. 4-6:
After the bacterial strain is colonized by the root system of the rice, the biomass of the rice and the nitrogen content of the overground part are increased, which indicates that the exogenously added bacterial strain promotes the growth of the rice to a certain extent in cadmium stress treatment. In the 0.3mg/L treatment group, the dry weight content of the plants increased from 41.37g to 52.64g, the total nitrogen content increased from 588.25mg to 652.25mg, and in the 2.0mg/L treatment group, the dry weight content of the plants increased from 27.43g to 34.50g, and the total nitrogen content increased from 432.47mg to 566.29mg.
After the strain is added, the cadmium content in the rice grains is obviously reduced, wherein in a 0.3mg/L treatment group, the cadmium content is reduced from 3.78mg/kg to 2.90mg/kg, the reduction rate is 23 percent, and in a 2.0mg/L treatment group, the cadmium content is reduced from 16.00mg/kg to 10.90mg/kg, and the reduction rate is 32 percent.
The invention can be seen that the discovered Burkholderia cepacia (Burkholderia cepacia) WW13 is a high-efficiency cadmium-resistant strain. The strain has certain application potential in efficiently adsorbing free cadmium ions, promoting rice growth and nitrogen absorption, reducing cadmium accumulation in seeds and ensuring safe production and utilization of agricultural products, and other similar genus has weaker capability.
It should be noted that: burkholderia ZCC was tested according to the method of the invention described above, with no significant difference in overall nitrogen content from the control in the 2.0mg/L treated group.
Finally, it should also be noted that the above list is merely a few specific embodiments of the present invention. Obviously, the invention is not limited to the above embodiments, but many variations are possible. All modifications directly derived or suggested to one skilled in the art from the present disclosure should be considered as being within the scope of the present invention.
Claims (3)
1. Burkholderia cepacia (Burkholderia cepacia) WW13 characterized by a accession number: cgmccno.27717.
2. Use of burkholderia cepacia (Burkholderia cepacia) WW13 according to claim 1 to promote safe production of rice under cadmium stress.
3. Use according to claim 2, characterized in that: high tolerance and cadmium adsorption capacity, has the function of nitrogen fixation, can promote rice growth and nitrogen absorption, and reduces rice grain cadmium accumulation.
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