CN108160687B - Method for restoring uranium-polluted soil in arid region by using growth-promoting bacterium reinforced plants - Google Patents
Method for restoring uranium-polluted soil in arid region by using growth-promoting bacterium reinforced plants Download PDFInfo
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- CN108160687B CN108160687B CN201810127112.1A CN201810127112A CN108160687B CN 108160687 B CN108160687 B CN 108160687B CN 201810127112 A CN201810127112 A CN 201810127112A CN 108160687 B CN108160687 B CN 108160687B
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
- B09C1/105—Reclamation of contaminated soil microbiologically, biologically or by using enzymes using fungi or plants
Abstract
A method for restoring uranium contaminated soil in arid regions by using growth-promoting bacterium-enhanced plants comprises the steps of putting seeds of macleaya cordata into a sodium hypochlorite solution for soaking and disinfecting, washing the seeds with sterile water, directly inoculating Pseudomonas fluorescens, spraying Pseudomonas fluorescens suspension to the seeds in the germination process, transplanting germinated seedlings into pots inoculated with the Pseudomonas fluorescens suspension, cultivating and transplanting the seedlings into the uranium contaminated drought soil, pouring Pseudomonas fluorescens suspension to the plants, harvesting the plants after 150 days of 120-year storage, and carrying out centralized treatment. The method takes the macleaya cordata and the Pseudomonas fluorescens which are uranium-enriched plants as materials, solves the problem of environmental hazard of the uranium-polluted soil in the prior arid region, has the advantages of simple and convenient operation and management, high repair efficiency, low cost, small environmental risk, capability of beautifying the environment and the like compared with the prior technical method, and is particularly suitable for repairing the uranium-polluted soil around uranium-containing solid waste stacking places such as uranium mines, hydrometallurgy plants, uranium waste ore and tailing ponds and the like in the northern arid region of China.
Description
Technical Field
The invention relates to the technical field of uranium contaminated soil phytoremediation, in particular to a method for restoring uranium contaminated soil in arid regions by using growth-promoting bacteria reinforced plants.
Background
The uranium contaminated soil is mainly generated by human activities such as uranium mining and metallurgy, nuclear weapon tests, nuclear accident leakage and the like. Uranium is a radioactive element with strong toxicity, and the toxicity is mainly expressed as chemical toxicity and radioactive toxicity, so that the uranium is extremely harmful to human bodies. Uranium in the contaminated soil may enter the human body through the food chain, increasing the risk of cancer in the human body. The remediation of uranium-contaminated soil is related to the sustainable development of the nuclear industry and the health of human beings, and becomes an environmental problem to be solved urgently at present.
In recent years, plant restoration technology attracts attention by virtue of in-situ, economical, green and the like, and becomes an effective means for treating soil polluted by heavy metals and radionuclides. The effect of phytoremediation depends on the uranium enrichment capacity of the plant and the biomass of the plant. At present, most plants with ultra-enrichment or enrichment capacity to uranium have the problems of low biomass, limited plant root expansion depth, poor adaptability of plants to the environment and the like, so that how to increase the biomass of the plants and improve the resistance of the plants to the environment becomes a key technology for restoring uranium-polluted soil by plants. The plant growth-promoting rhizobacteria and the secretion thereof can not only provide necessary nutrient substances and growth regulating factors for plants to improve the biomass of the plants, but also obviously change the activity and transferability of uranyl ions in the polluted soil, thereby achieving the purpose of improving the plant repair efficiency.
The plant growth-promoting rhizobacteria can produce substances such as indoleacetic acid, siderophores, ACC deaminase, cyanic acid and the like which can promote the growth of plants, and help some plants to survive and grow under drought conditions. When the plant growth-promoting rhizobacteria has better stress resistance, the resistance of host plants to heavy metals, drought, salinity, plant pathogens and the like can be increased, so that the plant repair is more efficient. According to the current exploitation and utilization conditions of uranium ore resources in China, sandstone-type uranium ores become the main type of uranium ore resource exploitation and metallurgy in China, are mainly distributed in the north, have less rainfall and have relatively arid soil. When plants are used for repairing uranium-contaminated soil in arid regions, the drought resistance of the plants must be improved.
Disclosure of Invention
Aiming at the problems, the invention aims to provide a method for restoring uranium-polluted soil in arid regions by utilizing growth-promoting bacterium reinforced plants, which has the advantages of convenient material acquisition, low cost, high restoring efficiency, simple and convenient processing steps, small environmental risk and the like.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a method for restoring uranium contaminated soil in arid regions by using growth-promoting bacterium enhanced plants is characterized in that macleaya cordata is inoculated with Pseudomonas fluorescens bacteria, and germinated macleaya cordata seedlings are planted in the arid uranium contaminated soil to survive and grow, so that uranium is enriched in macleaya cordata plants.
The further technical scheme of the invention is as follows: soaking the macleaya cordata seeds in a sodium hypochlorite solution for disinfection, washing the macleaya cordata seeds clean by using sterile water, directly inoculating Pseudomonas fluorescens bacteria to the macleaya cordata, spraying Pseudomonas fluorescens bacterial suspension to the seeds in the germination process, transplanting the germinated seedlings into a pot inoculated with the Pseudomonas fluorescens bacterial suspension, transplanting the seedlings into uranium-polluted drought soil after cultivation, watering the plants with the Pseudomonas fluorescens bacterial suspension, harvesting the plants after 120-day heat treatment, and performing centralized treatment.
The further technical scheme of the invention is as follows: the specific method for the disinfection treatment of the macleaya cordata seeds comprises the following steps: soaking plump Macleaya cordata seeds in sterile water for 6-8h, taking out, draining, soaking in 2% sodium hypochlorite solution for 10-12min for sterilization, rinsing the seeds with sterile water for 3-5 times, each time for 0.5-1.5min, and spreading the seeds after sterilization.
The further technical scheme of the invention is as follows: the specific method for inoculating Pseudomonas fluorescens is as follows: inoculating Pseudomonas fluorescens strain into liquid culture medium containing peptone 3-8g/L (preferably 4-6g/L, more preferably 5g/L), beef extract 1-5g/L (preferably 2-4g/L, more preferably 3g/L), and sodium chloride 3-7g/L (preferably 4-6g/L, more preferably 5g/L), culturing in biochemical incubator at 28.0-28.5 deg.C, centrifuging the cultured bacteria solution at 8000r/min for 5-6min, collecting centrifuged bacteria, and diluting OD of the bacteria suspension with sterile water600Is 0.6 to 1.5 (preferably 0.6 to 1.5, more preferably 1.0) (OD)6001.0≈108CFU/mL), add 100-120mL OD to the petri dish containing sterilized macleaya cordata seeds600The infection with the suspension of Pseudomonas fluorescens is 0.6-1.5 (preferably 0.6-1.5, more preferably 1.0) for 2-3 h.
The further technical scheme of the invention is as follows: the specific method for seed germination is as follows: draining the infected seeds in a sterile operating platform, sowing the seeds into germination boxes, sowing 10-30 seeds (preferably 15-25 seeds, more preferably 20 seeds) in each hole, placing the germination boxes in a light incubator, controlling the temperature at 20-30 ℃ (preferably 22-28 ℃, more preferably 25-26 ℃) and the relative humidity at 50-70% (preferably 55-68%, more preferably 60-65%), carrying out light treatment for 8-16h (preferably 10-14 ℃, more preferably 12h), carrying out dark treatment for 8-16h (preferably 10-14 ℃, more preferably 12h), spraying the seeds with Pseudomonas fluorescens suspension for 1-2 times every 20-30h (preferably 22-26 ℃, more preferably 24h) during germination, spraying 5-8mL (preferably 6-7mL) of each germination box, the seedling is broken, and is regarded as germination, and the germination time is 15-25d (preferably 18-22d, more preferably 20 d).
The further technical scheme of the invention is as follows: the specific method for cultivating and transplanting the seedlings comprises the following steps: crushing soil, naturally drying, sieving by a 4mm sieve, and mixing with river sand with the particle size of 0.25-0.35mm according to the weight ratio of 2-4: 1, continuously sterilizing with high pressure steam for 2-4 days, placing the soil in a greenhouse with temperature of 20-30 deg.C (preferably 22-28 deg.C, more preferably 25-26 deg.C) and humidity of 50-65% (preferably 52-62%, more preferably 55-60%), and mixing to obtain the final product6000.6-1.5 (preferably 0.6-1.5, more preferably 1.0) of Pseudomonas fluorescens suspension is poured into sterilized soil, 150mL (preferably 110-140mL, more preferably 120mL) of the suspension is poured into each kilogram of soil, the mixture is stirred and mixed uniformly by a sterilizing shovel, the flowerpot inoculated with the Pseudomonas fluorescens suspension is sealed by a preservative film, the flowerpot is placed in a greenhouse with the temperature of 20-30 ℃ (preferably 22-28 ℃, more preferably 25-26 ℃) for incubation, the seedlings with consistent growth in the germination box are transplanted into the flowerpot inoculated with the Pseudomonas fluorescens suspension, after the seedlings grow to 25-35d, the seedlings are transplanted into the soil polluted by uranium, the Pseudomonas fluorescens suspension is poured into the flowerpot once every 10-20d, 300mL (preferably 220-280 mL) of the Pseudomonas fluorescens suspension is poured into each plant once (preferably 220-150 mL, more preferably 120mL) of the flowerpot, more preferably 100- 120d) followed by harvesting of the plant.
The specific method for harvesting plants and carrying out centralized treatment comprises the following steps: the method is characterized in that plants are harvested manually or mechanically, the harvested plants are transferred to a safe place to be concentrated and dried, smashed, burnt and finally buried or leached to recover uranium, and uranium-polluted soil in arid regions is effectively restored.
The plant repairing method for uranium-polluted soil in the arid region takes uranium-enriched plants macleaya cordata and Pseudomonas fluorescens as materials, solves the problem of environmental hazard of the uranium-polluted soil in the prior arid region, and has the following technical advantages compared with the prior art;
(1) after Pseudomonas fluoscens are inoculated in the rhizosphere of the macleaya cordata, the biomass, the uranium enrichment capacity and the drought resistance capacity of the macleaya cordata are obviously improved, a good effect is achieved on the remediation of uranium-polluted soil in drought regions, and plants can be harvested manually or mechanically and transferred to a safe place for processing.
(2) The method adopts the macleaya cordata inoculated with Pseudomonas fluorescens to carry out phytoremediation on the uranium contaminated soil in the arid region, can greatly reduce the uranium from entering underground water and surrounding farmlands, can cultivate plants in the uranium contaminated soil in the arid region, can fix the uranium contaminated soil, reduce dust emission and reduce the diffusion of uranium pollutants.
(3) Macleaya cordata is not ingested by cattle and sheep, and therefore uranium is not transmitted through the food chain to increase environmental risk.
(4) The pink plumepoppy inoculated with Pseudomonas fluorescens can be directly planted in uranium contaminated soil in arid regions, and has the advantages of simple and convenient operation and management, high remediation efficiency, low cost, small environmental risk, capability of beautifying the environment and the like.
The method is suitable for repairing the uranium polluted soil around uranium-containing solid waste stacking places such as uranium mines, hydrometallurgy plants, uranium waste ore and tailing ponds and the like in northern arid regions of China.
Detailed Description
The present invention will be further described with reference to the following embodiments.
Example 1:
a method for restoring uranium contaminated soil in arid regions by using growth-promoting bacterium-enhanced plants comprises the steps of putting seeds of macleaya cordata into a sodium hypochlorite solution for soaking and disinfecting, washing the seeds with sterile water, directly inoculating Pseudomonas fluorescens, spraying Pseudomonas fluorescens suspension to the seeds in the germination process, transplanting germinated seedlings into pots inoculated with the Pseudomonas fluorescens suspension, cultivating and transplanting the seedlings into the uranium contaminated drought soil, pouring Pseudomonas fluorescens suspension to the plants, harvesting the plants after 150 days of 120-year storage, and carrying out centralized treatment.
The specific method for the disinfection treatment of the macleaya cordata seeds comprises the following steps: soaking plump Macleaya cordata seeds in sterile water for 6h, taking out and draining, soaking the seeds in 2% sodium hypochlorite solution for 10min for disinfection, rinsing the seeds with sterile water for 3 times, each time for 0.6min, and spreading the seeds after the disinfection is finished.
The specific method for inoculating Pseudomonas fluorescens is as follows: inoculating Pseudomonas fluorescens strain into liquid culture medium containing peptone 4g/L4, beef extract 2g/L and sodium chloride 4g/L, culturing in biochemical incubator at 28.0-28.5 deg.C, centrifuging the cultured bacteria liquid at 8000r/min for 5-6min to obtain centrifuged bacteria, and diluting OD of the bacterial suspension with sterile water600To 0.8, 100mL OD was added to the petri dish containing sterilized macleaya cordata seeds600The infection with the suspension of Pseudomonas fluorescens was 0.8 for 2 h.
The specific method for seed germination is as follows: draining the infected seeds in a sterile operation table, sowing the seeds into germination boxes, sowing 15 seeds in each hole, sowing 10 holes in each group, placing the germination boxes in an illumination incubator, controlling the temperature at 22 ℃, controlling the relative humidity at 50%, performing illumination treatment for 10h, performing dark treatment for 14h, spraying Pseudomonas fluorescens suspension to the seeds for 1 time every 22h in the germination process, spraying 5mL to each germination box, and regarding the seedlings as germination after breaking the soil, wherein the germination time is 18.
The specific method for cultivating and transplanting the seedlings comprises the following steps: crushing soil, naturally drying, sieving by a 4mm sieve, and mixing with river sand with the particle size of 0.25-0.35mm according to the weight ratio of 2: 1, continuously sterilizing the soil for 2 days by high-pressure steam, placing the soil in a greenhouse with the temperature of 22 ℃ and the humidity of 52 percent, and mixing the OD600Pouring 0.8 gram of Pseudomonas fluorescens bacterial suspension into sterilized soil, pouring 100-,transplanting the plants into uranium-polluted drought soil, watering the plants once every 12 days with 200-300mL (preferably 220-280mL, more preferably 240-260mL) of Pseudomonas fluorescens bacterial suspension once per plant, and harvesting the plants after 120-150d (preferably 110-140d, more preferably 100-120 d).
The specific method for harvesting plants and carrying out centralized treatment comprises the following steps: the method is characterized in that plants are harvested manually or mechanically, the harvested plants are transferred to a safe place to be concentrated and dried, smashed, burnt and finally buried or leached to recover uranium, and uranium-polluted soil in arid regions is effectively restored.
Through detection, the average content of uranium in roots of each plant is 425.58mg/kg, and the weight of root ash is 1.05 g; the content of uranium in the stems is 22.58mg/kg, and the weight of stem ash is 0.88 g; the content of uranium in the leaves is 55.25mg/kg, and the weight of the ash in the leaves is 2.15 g; on average each plant can be cleaned of 0.59mg uranium from drought soil with a relative humidity of 40%.
Directly planting macleaya cordata which is not inoculated with Pseudomonas fluorescens in drought soil with the relative humidity of 40%, harvesting the plants after 120 days, and detecting that the average content of uranium in roots of each plant is 278.55mg/kg and the weight of root ash is 0.75 g; the content of uranium in the stems is 12.64mg/kg, and the weight of stem ash is 0.45 g; the content of uranium in the leaves is 28.55mg/kg, and the weight of the ash in the leaves is 1.63 g; on average each plant can be cleaned of 0.26mg uranium from drought soil with a relative humidity of 40%.
Therefore, the repair efficiency of the macleaya cordata planted with Pseudomonas fluorescens in arid soil with the relative humidity of 40% is improved by 127% compared with the macleaya cordata planted with non-Pseudomonas fluorescens.
Example 2:
a method for restoring uranium contaminated soil in arid regions by using growth-promoting bacterium-enhanced plants comprises the steps of putting seeds of macleaya cordata into a sodium hypochlorite solution for soaking and disinfecting, washing the seeds with sterile water, directly inoculating Pseudomonas fluorescens, spraying Pseudomonas fluorescens suspension to the seeds in the germination process, transplanting germinated seedlings into pots inoculated with the Pseudomonas fluorescens suspension, cultivating and transplanting the seedlings into the uranium contaminated drought soil, pouring Pseudomonas fluorescens suspension to the plants, harvesting the plants after 150 days of 120-year storage, and carrying out centralized treatment.
The specific method for the disinfection treatment of the macleaya cordata seeds comprises the following steps: soaking plump Macleaya cordata seeds in sterile water for 8h, taking out and draining, soaking the seeds in 2% sodium hypochlorite solution for 12min for disinfection, rinsing the seeds with sterile water for 5 times, 1.2min each time, and spreading the seeds after the disinfection is finished.
The specific method for inoculating Pseudomonas fluorescens is as follows: inoculating Pseudomonas fluorescens strain into liquid culture medium containing peptone 7g/L, beef extract 4g/L and sodium chloride 6g/L, culturing in biochemical incubator at 28.0-28.5 deg.C, centrifuging the cultured bacterial liquid at 8000r/min for 5-6min, collecting centrifuged bacteria, and diluting OD of bacterial suspension with sterile water600At 1.2, the dishes containing the sterilized seeds of Macleaya cordata were infected with 120mL of Pseudomonas fluorescens suspension at OD600 of 1.2 for 3 h.
The specific method for seed germination is as follows: draining the infected seeds in a sterile operation table, sowing the seeds into germination boxes, sowing 25 seeds in each hole, sowing 10 holes in each group, placing the germination boxes in an illumination incubator, controlling the temperature at 28 ℃, controlling the relative humidity at 68%, carrying out illumination treatment for 14h, carrying out dark treatment for 10h, spraying Pseudomonas fluorescens suspension for 2 times every 28h in the germination process, spraying 7mL in each germination box, and regarding the seedling as germination after breaking the soil, wherein the germination time is 22 d.
The specific method for cultivating and transplanting the seedlings comprises the following steps: crushing soil, naturally drying, sieving by a 4mm sieve, and mixing with river sand with the particle size of 0.25-0.35mm according to the weight ratio of 4: 1, continuously sterilizing for 4 days with high pressure steam, placing the soil in a greenhouse with a temperature of 28 ℃ and a humidity of 62%, and mixing the OD600Pouring 1.2 of Pseudomonas fluorescens bacterial suspension into sterilized soil, pouring 140mL of bacterial suspension into each kilogram of soil, uniformly mixing by using a sterilizing shovel, sealing a flowerpot inoculated with the Pseudomonas fluorescens bacterial suspension by using a preservative film, placing the flowerpot in a greenhouse at 28 ℃ for incubation for three weeks, and transplanting seedlings with consistent growth vigor in a germination box to the sterilized soilInoculating the Pseudomonas fluoescens suspension into the flowerpot, transplanting the flowerpot into uranium-polluted drought soil after the seedlings grow to 32d, watering the plants once every 18d, watering 280mL of Pseudomonas fluoescens suspension once every plant, and harvesting the plants after 140 d.
The specific method for harvesting plants and carrying out centralized treatment comprises the following steps: the method is characterized in that plants are harvested manually or mechanically, the harvested plants are transferred to a safe place to be concentrated and dried, smashed, burnt and finally buried or leached to recover uranium, and uranium-polluted soil in arid regions is effectively restored.
Through detection, the average content of uranium in roots of each plant is 410.50mg/kg, and the weight of root ash is 0.98 g; the content of uranium in the stems is 23.55mg/kg, and the weight of stem ash is 0.55 g; the content of uranium in the leaves is 52.88mg/kg, and the weight of the ash in the leaves is 2.09 g; on average each plant can be cleaned of 0.52mg uranium from drought soil with a relative humidity of 30%.
Directly planting macleaya cordata which is not inoculated with Pseudomonas fluorescens in drought soil with the relative humidity of 30%, harvesting the plants after 120 days, and detecting that the average content of uranium in roots of each plant is 198.56mg/kg and the weight of root ash is 0.72 g; the content of uranium in the stems is 11.98mg/kg, and the weight of stem ash is 0.35 g; the content of uranium in the leaves is 25.62mg/kg, and the weight of the ash in the leaves is 1.42 g; on average, each plant can be cleaned of 0.18mg of uranium from drought soil with a relative humidity of 30%.
Therefore, the repair efficiency of the macleaya cordata planted with Pseudomonas fluorescens in arid soil with the relative humidity of 30% is improved by 189% compared with the macleaya cordata planted with non-Pseudomonas fluorescens.
Example 3:
a method for restoring uranium contaminated soil in arid regions by using growth-promoting bacterium-enhanced plants comprises the steps of putting seeds of macleaya cordata into a sodium hypochlorite solution for soaking and disinfecting, washing the seeds with sterile water, directly inoculating Pseudomonas fluorescens, spraying Pseudomonas fluorescens suspension to the seeds in the germination process, transplanting germinated seedlings into pots inoculated with the Pseudomonas fluorescens suspension, cultivating and transplanting the seedlings into the uranium contaminated drought soil, pouring Pseudomonas fluorescens suspension to the plants, harvesting the plants after 150 days of 120-year storage, and carrying out centralized treatment.
The further technical scheme of the invention is as follows: the specific method for the disinfection treatment of the macleaya cordata seeds comprises the following steps: soaking plump Macleaya cordata seeds in sterile water for 7h, taking out and draining, soaking the seeds in 2% sodium hypochlorite solution for 11min for disinfection, rinsing the seeds with sterile water for 4 times, 1.0min each time, and spreading the seeds after the disinfection is finished.
The specific method for inoculating Pseudomonas fluorescens is as follows: inoculating Pseudomonas fluorescens strain into liquid culture medium containing peptone 5g/L, beef extract 3g/L and sodium chloride 5g/L, culturing in biochemical incubator at 28.0-28.5 deg.C, centrifuging the cultured bacterial liquid at 8000r/min for 5-6min, collecting centrifuged bacteria, and diluting OD of the bacterial suspension with sterile water600At 1.0, the dishes containing the sterilized seeds of Macleaya cordata were infected with 110mL of Pseudomonas fluorescens suspension having an OD600 of 1.0 for 2.5 h.
The specific method for seed germination is as follows: draining the infected seeds in a sterile operation table, sowing the seeds into germination boxes, sowing 20 seeds in each hole, sowing 10 holes in each group, placing the germination boxes in an illumination incubator, controlling the temperature at 25-26 ℃, controlling the relative humidity at 60-65%, performing illumination treatment for 12h, performing dark treatment for 12h, spraying Pseudomonas fluorescens suspension to the seeds every 24h during germination, spraying 6-7mL to each germination box, and regarding the seedlings as germination after breaking the soil, wherein the germination time is 20 d.
The specific method for cultivating and transplanting the seedlings comprises the following steps: crushing soil, naturally drying, sieving by a 4mm sieve, and mixing with river sand with the particle size of 0.25-0.35mm according to the weight ratio of 3: 1, continuously sterilizing the soil for 3 days by high-pressure steam, placing the soil in a greenhouse with the temperature of 25-26 ℃ and the humidity of 55-60%, and mixing the OD600Pouring 1.0 of Pseudomonas fluorescens bacterial suspension into sterilized soil, pouring 130mL of bacterial suspension per kilogram of soil, stirring and mixing uniformly by using a sterilizing shovel, sealing a flowerpot inoculated with the Pseudomonas fluorescens bacterial suspension by using a preservative film, and placing the flowerpot in a greenhouse at 25-26 DEG CIncubating for three weeks, transplanting the seedlings with consistent growth vigor in the germination boxes into flowerpots inoculated with the Pseudomonas fluorescens bacterial suspension, transplanting the seedlings into uranium-polluted drought soil after the seedlings grow to 30d, watering the plants once every 15d with the Pseudomonas fluorescens bacterial suspension, watering 250mL of each plant once, and harvesting the plants after 140 d).
The specific method for harvesting plants and carrying out centralized treatment comprises the following steps: the method is characterized in that plants are harvested manually or mechanically, the harvested plants are transferred to a safe place to be concentrated and dried, smashed, burnt and finally buried or leached to recover uranium, and uranium-polluted soil in arid regions is effectively restored.
Through detection, the average content of uranium in roots of each plant is 388.55mg/kg, and the weight of root ash is 0.95 g; the content of uranium in the stems is 20.98mg/kg, and the weight of stem ash is 0.52 g; the content of uranium in the leaves is 51.36mg/kg, and the weight of the ash in the leaves is 1.95 g; on average, each plant was able to remove 0.48mg of uranium from arid soil with a relative humidity of 20%.
Directly planting macleaya cordata which is not inoculated with Pseudomonas fluorescens in drought soil with the relative humidity of 20%, harvesting the plants after 120 days, and detecting that the average content of uranium in roots of each plant is 158.35mg/kg and the weight of root ash is 0.59 g; the content of uranium in the stems is 10.54mg/kg, and the weight of stem ash is 0.32 g; the content of uranium in the leaves is 22.58mg/kg, and the weight of the ash in the leaves is 1.26 g; on average, each plant was able to remove 0.13mg of uranium from arid soil with a relative humidity of 20%.
Therefore, the repair efficiency of the macleaya cordata planted with Pseudomonas fluorescens in arid soil with the relative humidity of 20% is improved by 269% compared with the macleaya cordata planted with non-Pseudomonas fluorescens.
The above is only a preferred embodiment of the present invention, and various modifications and changes can be made thereto by those skilled in the art based on the above-described concept of the present invention. For example, the method is planted in drought soil with different uranium concentrations, planted in uranium polluted soil with different water contents, and adopted with different harvesting modes and periods, different seedling raising methods and the like. However, such similar changes and modifications are also within the spirit of the present invention.
Claims (5)
1. A method for restoring uranium contaminated soil in arid regions by using growth-promoting bacterium-enhanced plants is characterized in that macleaya cordata seeds are placed in a sodium hypochlorite solution for soaking and disinfection, are washed clean by sterile water, are directly inoculated with Pseudomonas fluorescens, are sprayed with Pseudomonas fluorescens suspension in the germination process, are transplanted into pots inoculated with Pseudomonas fluorescens suspension, are cultivated and transplanted into uranium contaminated drought soil, are watered with corresponding Pseudomonas fluorescens suspension, are alive and grow, enrich uranium in macleaya cordata plants, and are harvested after 150 days of 120 + and concentrated;
the specific method for sterilizing the macleaya cordata seeds comprises the following steps: soaking plump Macleaya cordata seeds in sterile water for 6-8h, taking out, draining, soaking in 2% sodium hypochlorite solution for 10-12min for sterilization, rinsing the seeds with sterile water for 3-5 times, each time for 0.5-1.5min, and spreading the seeds after sterilization;
the specific method for inoculating Pseudomonas fluorescens is as follows: inoculating Pseudomonas fluorescens strain into liquid culture medium containing peptone 3-8g/L, beef extract 1-5g/L, and sodium chloride 3-7g/L, culturing in biochemical incubator at 28.0-28.5 deg.C, centrifuging the cultured bacteria liquid at 8000r/min for 5-6min to obtain centrifuged bacteria, and diluting OD of the bacterial suspension with sterile water6000.6-1.5, adding 120mL OD of 100-600The Pseudomonas fluorescens suspension with the infection time of 0.6-1.5 is infected for 2-3 h.
2. The method for restoring uranium-polluted soil in arid regions by using growth-promoting bacteria enhanced plants according to claim 1, which is characterized in that: the specific method for the germination of the macleaya cordata seeds comprises the following steps: draining the infected seeds in a sterile operation table, sowing the seeds into a germination box, sowing 10-30 seeds in each hole, placing the germination box in an illumination incubator, controlling the temperature at 20-30 ℃, controlling the relative humidity at 50-70%, carrying out illumination treatment for 8-16h and carrying out dark treatment for 8-16h, spraying Pseudomonas fluoroscein suspension for 1-2 times every 20-30h in the germination process, spraying 5-8mL in each germination box, and regarding the seedlings as germinated when the seedlings break the soil, wherein the germination time is 15-25 d.
3. The method for restoring uranium-contaminated soil in arid regions by using growth-promoting bacteria enhanced plants according to claim 1 or 2, which is characterized in that: the specific method for culturing and transplanting the macleaya cordata seedlings comprises the following steps: crushing soil, naturally drying, sieving by a 4mm sieve, and mixing with river sand with the particle size of 0.25-0.35mm according to the weight ratio of 2-4: 1, continuously sterilizing for 2-4 days with high pressure steam, placing the soil in a greenhouse with temperature of 20-30 deg.C and humidity of 50-65%, and mixing the obtained product with OD600Pouring 0.6-1.5 of Pseudomonas fluorescens bacterial suspension into sterilized soil, pouring 100-150mL of Pseudomonas fluorescens bacterial suspension per kilogram of soil, stirring and mixing uniformly by using a sterilizing shovel, sealing a flowerpot inoculated with the Pseudomonas fluorescens bacterial suspension by using a preservative film, placing the flowerpot into a greenhouse at 20-30 ℃ for hatching, transplanting seedlings with consistent growth vigor in a germination box into the flowerpot inoculated with the Pseudomonas fluorescens bacterial suspension, transplanting the seedlings into uranium-polluted drought soil after the seedlings grow to 25-35d, pouring 200-300mL of Pseudomonas fluorescens bacterial suspension once every 10-20d, and harvesting the Plumbum zone after each Plumbum zone reaches 120-150 d.
4. The method for restoring uranium-contaminated soil in arid regions by using growth-promoting bacteria enhanced plants according to claim 1 or 2, which is characterized in that: the specific method for harvesting the macleaya cordata and carrying out centralized treatment comprises the following steps: the macleaya cordata is harvested manually or mechanically, the harvested macleaya cordata is moved to a safe place of the macleaya cordata to be concentrated and dried, smashed and burnt, and finally, the uranium is buried or leached to recover the uranium, so that the uranium-polluted soil in the arid area is effectively repaired.
5. The method for restoring uranium-polluted soil in arid regions by using growth-promoting bacteria enhanced plants according to claim 3, which is characterized in that: the specific method for harvesting the macleaya cordata and carrying out centralized treatment comprises the following steps: the macleaya cordata is harvested manually or mechanically, the harvested macleaya cordata is moved to a safe place of the macleaya cordata to be concentrated and dried, smashed and burnt, and finally, the uranium is buried or leached to recover the uranium, so that the uranium-polluted soil in the arid area is effectively repaired.
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