CN102936574A - Heavy metal resistant nodule bacterium and method of promoting tailings area plant restoration by using same - Google Patents
Heavy metal resistant nodule bacterium and method of promoting tailings area plant restoration by using same Download PDFInfo
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Abstract
The invention belongs to the technical field of agriculture and environmental pollution improvement and relates to a heavy metal resistant plant endogenous nodule bacterium and application thereof. The heavy metal resistant nodule bacterium is preserved at the China center for type culture collection (CCTCC) with the preservation date to be 2012/9/18 and culture preservation number to be CCTCC NO:M2012357. The bacterial strain liquid preparation contains more than a billion of effective living bacteria per milliliter, and the bacterial strain solid preparation contains 0.2 billion of effective living bacteria per gram. The heavy metal resistant nodule bacterium strain can secrete plant hormones indole acetic acid (IAA) and siderophores, and resists heavy metal copper and cadmium. When plants are planted in moist soil or tailings containing heavy metal and bioremediation preparation is inoculated, the heavy metal resistant nodule bacterium can promote plant growth and absorbs heavy metal copper, can improve plant restoration efficiency, can fix sandy soil and reduces water and soil loss.
Description
Technical field
The invention belongs to agricultural and environmental pollution treatment technology field, relate to a kind of root nodule bacterium of anti-heavy metal and the method for promotion mine tailing district plant-growth thereof.
Background technology
Mining, smelting process have brought that ecoscape goes to pot, Soil structure variation, nutrient and water lack, species diversity falls sharply, the waste (CHARACTERISTICS OF TAILINGS SAND, ore etc.) produced needs the large-area place of banking up, the abandoned land vegetation can not recover in time, many ecological environment problems such as serious soil erosion and heavy metal contamination, thus the havoc of mining area land ecosystem caused.It is reported, since the nineties in 20th century, China's Abandoned Land of Mine ecological recovery rate only 8.75%.Therefore, improvement and the Revegetation of reinforcement Mining Wasteland have become one of urgent task that China is current faced.
The mine tailing district, many tailings, be often uneven, irregular sand drift, and viscosity is poor, water-permeable is strong, corrosion stability is little, preserve moisture and fertility power is poor, and the essential organic matter of plant-growth and n p k nutrition component content are all relatively low, are unfavorable for the growth of plant.Plant-growth promoting rhizobacteria is the useful microbe groups that plant obtains in very long evolutionary process, plant is resisted to extraneous poor environment and have vital role.The scientific research personnel has started the revegetation for abandoned land and contaminated site by plant-growth promoting rhizobacteria both at home and abroad.After Bashan etc. find inoculation Azospirillum brasilense, be transplanted to three kinds of Root and stem of Cholla growths on urban waste ground and accelerate.Some bacterial classifications of the rhizobium (Rhizobium) of the discovery such as Wei and leguminous plants endosymbiosis can produce plant hormone and promote soybeans they grow, and have improved the absorption of soybean to heavy metal.Petrisor etc. and Wu etc. have started to explore PGPB have been applied in to mine tailing plant stability aspect.Therefore, utilize plant-growth promoting rhizobacteria to improve phytoremediation efficiency, soil erosion and heavy metal contamination that the wind erosion of control Mine Abandoned Land and water erosion cause, will be a kind of effective, eco-friendly reparation means.
Summary of the invention
The objective of the invention is the above-mentioned deficiency for prior art, a kind of root nodule bacterium of anti-heavy metal are provided.
Another object of the present invention is to provide the biological restoration preparation that contains this bacterium.
Another purpose of the present invention is to provide the application of this bacterium and biological restoration preparation.
Purpose of the present invention can be achieved through the following technical solutions:
A kind of endophytic bacterium W33, Classification And Nomenclature is Rhizobium sp.W33, is preserved in Chinese Typical Representative culture collection center, and preservation date is on September 18th, 2012, and culture presevation number is CCTCC NO:M 2012357.This endophytic bacterium W33 separates and obtains from Semen Euphorbiae (Leptochloa chinensis (Linn.) Nees) rhizosphere soil of Longshan, NanJing City, Jiangsu Province,China growth, through being accredited as Rhizobium sp..Main Biological is: G
-, during young age, thalline is shaft-like, without gemma.Well-grown on YMA medium, single bacterium colony circle, neat in edge, diameter 1～3mm, white is opaque.Glucose, sucrose, fructose, pectinose, wood sugar, N.F,USP MANNITOL, sorbyl alcohol etc. can be utilized, starch, gelatin etc. can not be utilized.
A kind of biological restoration preparation that contains the endophytic bacterium W33 that described preserving number is CCTCC NO:M 2012357.
Described biological restoration preparation is preferably liquid preparation, and the W33 bacterial strain living bacteria count that wherein to contain preserving number be CCTCC NO:M 2012357 is more than 1,000,000,000/milliliter.
Described liquid preparation is to prepare by conventional liquid submerged fermentation method fermentation W33 bacterial strain.
Described biological restoration preparation also can be preferably solid preparation, the W33 bacterial strain living bacteria count that wherein to contain preserving number be CCTCC NO:M 2012357 be 200,000,000/more than gram.
Described solid preparation is to use the W33 bacterial strain fermentation liquor that the peat composed of rotten mosses, peat or other carrier adsorption preserving numbers are CCTCC NO:M 2012357 to be mixed with granule or pulvis.
The application of the W33 bacterial strain that preserving number of the present invention is CCTCC NO:M 2012357 in mine tailing district phytoremediation.
The application of W33 bacterial strain in the phytoremediation of heavy-metal contaminated soil that preserving number of the present invention is CCTCC NO:M 2012357.
The application of biological restoration preparation in the phytoremediation of mine tailing district or heavy-metal contaminated soil of W33 bacterial strain of the present invention.
A kind of described biological restoration preparation containing the W33 bacterial strain is used for the plant restoration method of heavy metal pollution of soil: sow plant seed in moistening soil or mine tailing district containing heavy metal, inoculation W33 bacterial strain biological restoration preparation, every mu of inoculation 10
8The biological restoration preparation 4-6kg of individual bacterium/mL, minute 1-2 inoculation.
Beneficial effect
Endophytic bacterium W33 bacterial strain of the present invention can tolerate various heavy (minimum inhibition concentration of Cu, Pb, Cd be respectively 0.8,3.6,0.4mM).
The method of plant-growth promoting rhizobacteria of the present invention and the reparation of fortification of plants heavy metal pollution of soil thereof compared with prior art has the following advantages:
(1) W33 bacterial strain of the present invention can produce indolylacetic acid, and the IAA production peak can reach 16mgL
-1There is molten phosphorus characteristic, help lend some impetus to plant-growth.The high yield siderophore, contribute to improve stress resistance of plant (drought resisting, waterlogging-resistant, anti-saline and alkaline, disease and insect resistance).
(2) utilize the plant-growth promoting rhizobacteria Promoting plant growth, improve plant heavy metal Cu content, fortification of plants extracts remediation efficiency.
(3) fixing sand, reduce soil erosion.
Biological sample preservation information
Plant-growth promoting rhizobacteria W33, Classification And Nomenclature is Rhizobium sp.W33, is preserved in Chinese Typical Representative culture collection center, and the preservation address is Wuhan, China Wuhan University, and preservation date is on September 18th, 2012, and culture presevation number is CCTCC NO:M2012357.
Embodiment
Embodiment 1
The separation and purification from Semen Euphorbiae (Leptochloa chinensis (Linn.) Nees) rhizosphere soil in Longshan, Nanjing of the plant-growth promoting rhizobacteria W33 bacterial strain of the anti-heavy metal of the present invention obtains, and isolation and identification method is as follows:
Take the rhizosphere soil of close attachment at 2mm left and right, root system surface soil with aseptic nipper, 10g weighs, be placed in the triangular flask that fills the 90ml sterilized water, 30min fully vibrates, standing 10min, get the 0.1ml suspension, adopts 10 times of serial dilution methods, coat containing 0.004% Congo red YMA dull and stereotyped (sucrose 10.0g, MgSO
47H
2O 0.2g, K
2HPO
40.5g, NaCl 0.1g, CaCO
33.0g, yeast extract paste 5g, water 1000mL, agar 20g), cultivate 3d for 28 ℃, picking list bacterium colony is rule after purifying and is preserved on the YMA flat board.By inoculation in not adding CaCO
3The YMA nutrient solution in, 28 ℃ of shaking culture 18h, get 1.5mL bacterium liquid in the Eppendorf centrifuge tube, the centrifugal 5min of 5000rpm collects thalline, extract according to a conventional method bacteria total DNA, pcr amplification bacterial 16 S rDNA, amplified production through the order-checking after with GenBank in known 16S rDNA sequence compare, reach 99.5% with the 16S rDNA sequence similarity of Rhizobium lusitanum, it is accredited as to this genus.
Embodiment 2
By W33(CCTCC NO:M 2012357) streak inoculation is in the YMA medium containing different heavy metal concentrations, and can 30 ℃ of cultivation 3d, observe it and grow and growing state.The results are shown in Table 1.The W33 bacterial strain can tolerate various heavy (minimum inhibition concentration of Cu, Pb, Cd be respectively 0.8,3.6,0.4mM).
Table 1 bacterial strain is at the growing state containing on the YMA medium of different heavy metal concentrations
In upper table, "+" means growth, and "-" means not grow.
Embodiment 3
Measure siderophore according to the method for king's equality.By W33(CCTCC NO:M 2012357) be inoculated in the YMA liquid nutrient medium 30 ℃ of 150rpm shaking culture 48h.The centrifugal 5min of fermented liquid 12000rpm, get supernatant liquor and equal-volume CAS detection liquid fully mixes, and measures the light absorption value (A) at 630nm wavelength place after colour developing 1h, with deionized water, compares zeroing.Separately with the aseptic YMA liquid nutrient medium of equal-volume and CAS, detect the processing that liquid fully mixes, be measured in the same method light absorption value and be reference value (Ar).A/Ar value<1, can be considered to the high yield siderophore.Result shows W33(CCTCC NO:M 2012357) A/Ar of fermented liquid is 0.4, the high yield siderophore.
Embodiment 4
According to the measuring method of Gordon and Weber (1951), with test tube packing YMA liquid nutrient medium, every pipe 4mL, add the 2.5mg/mL tryptophane solution 1mL of filtration sterilization after 121 ℃ of high-temperature sterilizations, and the final concentration that makes tryptophane in substratum is 0.5mg/mL.By W33(CCTCC NO:M 2012357) be inoculated in above-mentioned substratum 30 ℃ of shaking table shaking culture 3d.Fermented liquid, in the centrifugal 5min of 12000r/min, is got supernatant liquor 1mL, adds the ortho-phosphoric acid 50 μ L of 10mM, then adds the 2mLSackowski's developer, fully mixes, and dark lower 25 ℃ of colour developing 30min measure light absorption value under the 530nm wavelength.Aseptic culture medium is the same to be done identical processing and returns to zero in contrast.Take concentration as 0,5,10,20,40, the IAA reference liquid of 60mg/L makes typical curve with method, calculates the concentration of IAA in fermented liquid.Result shows W33(CCTCC NO:M 2012357) can produce indolylacetic acid, the IAA production peak can reach 16mg/L.
The ability of embodiment 5 bacterial strain W33 dissolving phosphoric acid DFPs
Pick the W33(CCTCC NO:M 2012357 that cultivates the 18-24h gained in the YMA nutrient solution with transfering loop) bacterium liquid, at inorganic phosphorus substratum (glucose 10.0g, (NH
4)
2SO
40.5g, NaCl 0.3g, KCl 0.3g, MgSO
47H
2O 0.3g, FeSO
47H
2O0.03g, MnSO
4H
2O 0.03g, Ca
3(PO
4)
25.0g, H
2O 1000ml, pH 7.0-7.5,115 ℃ of sterilizing 30min) upper line, after cultivating 5d, observe periphery of bacterial colonies and have or not transparent molten phosphorus circle.Measure the ratio of molten phosphorus loop diameter and colony diameter.Result shows, bacterial strain W33(CCTCC NO:M 2012357) at inorganic phosphorus substratum well-grown, periphery of bacterial colonies produces transparent circle, can dissolve Ca in substratum
3(PO
4)
2Precipitation, the ratio of molten phosphorus loop diameter and colony diameter is 1.6 ± 0.2.
Embodiment 6 liquid preparations
W33(CCTCC NO:M 2012357 by YMA solid medium cultivation 48h gained) slant strains is inoculated in YMA medium (sucrose 10.0g, MgSO
47H
2O 0.2g, K
2HPO
40.5g, NaCl 0.1g, CaCO
33.0g, yeast extract paste 5g, water 1000mL, water 1000mL, pH7.0-7.5) in, cultivate after 24 hours and access seeding tank.Seeding tank is 0.5 ton, and the seed tank culture based component is sucrose 3.0kg, starch 0.6kg, (NH
4)
2SO
41.5kg, K
2HPO
40.3kg, MgSO
40.18kg, NaCl0.07kg, CaCO
30.4kg, yeast extract paste 0.2kg, 0.3 ton, water, amylase 5400U, soybean oil 50mL.Seeding tank must be first with steam sterilizing and be cooled to 28-30 ℃, inoculum size is that volume ratio 5%(be take culture volume as benchmark), the seed tank culture temperature is controlled at 28-32 ℃, stirring velocity is 220 rev/mins, the sterile air intake is 1: 0.8, cultivates after 20 hours seed liquor is all accessed to the production tank.Producing tank is 7 tons, and charging capacity is about 4.5 tons, and medium component is sucrose 40kg, starch 8kg, (NH
4)
2SO
415kg, KH
2PO
44.0kg, MgSO
42.4kg, NaCl 0.9kg, CaCO
34kg, yeast extract paste 2kg, 4.0 tons, water, amylase 64000U, soybean oil 400mL.Produce tank in advance with steam sterilizing and be cooled to 28-30 ℃, culture temperature is controlled at 28-32 ℃, and stirring velocity is 200 rev/mins, and the sterile air intake is 1: 0.8.More than after cultivation finishes, in nutrient solution, thalline quantity reaches 1,000,000,000/ml.It is biological preparation for repairing that nutrient solution gets final product filling.
The present embodiment is a kind of biological restoration formulation preparation method of suggestion, in fact biological restoration preparation of the present invention can prepare by conventional Rhizobium sp. microorganism belonging to genus fermentation culture method, as long as more than in nutrient solution, thalline quantity reaches 1,000,000,000/ml.
Embodiment 7 pulvis
The nutrient solution of embodiment 6 gained is adsorbed with the peat composed of rotten mosses, and nutrient solution is 1 with the ratio of the peat composed of rotten mosses: 3.5(L:Kg), be uniformly mixed, pulverize, obtain containing W33(CCTCC NO:M 2012357) pulvis.After testing, W33(CCTCC NO:M 2012357 in this solid biologic preparation for repairing) the bacterial strain living bacteria count be 200,000,000/more than gram.
Embodiment 8W33 inoculation Indian mustard extracts the copper in soil
Gather heavy metal cuprum polluted soil, air-dry grinding, the plastic tub of packing into alms bowl, every basin 2kg, adding water, to make water content be 60% of field capacity, keeps 2d, every basin is sown into 7, Indian mustard seed.Select commercially available Indian mustard Mix, Sin, tri-kinds of Pur.Inoculation embodiment 6 gained take the biological restoration microbial inoculum 10ml that W33 bacterial strain (CCTCC NO:M 2012357) be effective constituent, contrast and connect the equivalent sterilized water.During plant strain growth, add distilled water with weighting method every day, and to keep soil humidity be field capacity 60%.What growth inoculated embodiment 6 gained after 30d take biological restoration microbial inoculum 10ml that the W33 bacterial strain is effective constituent once.Sowing 60d results along native face clip plant overground part, wash out root system simultaneously, under 105 ℃, complete, 70 ℃ of oven dry, the dry weight of weighing root and overground part, plant sample grinds and with nitric acid-perchloric acid method, disappears and boil afterwards, and atomic absorption spectrophotometer is measured heavy metal Cu content in plant.The biomass that connects as shown in Table 2 the rear Indian mustard of bacterium W33 processing all improves than contrasting, wherein significantly increase by 80% and 73%, Pur kind plant shoot and the remarkable increase by 64% and 58% of root dry weight of the plant shoot of Mix kind and root dry weight.Sin breed organism amount increases minimum.As shown in Table 3, Indian mustard Mix and Pur kind Cu content are higher, absorb heavy metal energy force rate Sin strong, and after meeting bacterium W33 and processing, in Mix and Pur kind, Cu content increases lessly, and in the Sin kind, Cu content significantly improves and increases 31.6%-65.4%.As shown in Table 4, the Indian mustard of three kinds is energy enrichment Cu all, and after connecing the bacterium processing, the overground part of Indian mustard and root all increase than contrast to the enriching quantity of Cu, and wherein Sin kind overground part significantly increases by 1.03 times to the enriching quantity of Cu, is secondly the Pur kind.
The growgh promoting effects of root nodule bacterium W33 to Indian mustard in table 2 heavy metal Cu contaminated soil
| Plant variety | Project | Connect bacterium | Contrast |
| Mix | Overground part dry weight (g/ basin) | 2.26±0.02 | 1.25±0.03 |
| ? | Root heavy (g/ basin) | 0.45±0.01 | 0.26±0.03 |
| Sin | Overground part dry weight (g/ basin) | 1.90±0.32 | 1.81±0.16 |
| ? | Root heavy (g/ basin) | 0.58±0.02 | 0.42±0.04 |
| Pur | Overground part dry weight (g/ basin) | 2.66±0.33 | 1.62±0.12 |
| ? | Root heavy (g/ basin) | 0.52±0.17 | 0.33±0.02 |
The impact of root nodule bacterium W33 on Indian mustard Cu content in table 3 heavy-metal contaminated soil
| Plant variety | Project | Connect bacterium | Contrast |
| Mix | Overground part Cu content (mg/kg) | 90.8±4.2 | 97.5±18.3 |
| ? | Root Cu content (mg/kg) | 485.9±16.5 | 493.7±25.9 |
| Sin | Overground part Cu content (mg/kg) | 65.6±5.7 | 49.8±1.9 |
| ? | Root Cu content (mg/kg) | 449.9±2.4 | 272.0±1.3 |
| Pur | Overground part Cu content (mg/kg) | 89.0±2.3 | 92.4±3.9 |
| ? | Root Cu content (mg/kg) | 536.8±10.3 | 491.8±4.9 |
Table 4 heavy metallic activation bacterium strengthening Indian mustard extracts the copper in soil
| Plant variety | Project | Connect bacterium | Contrast | Increment rate % |
| Mix | Overground part Cu enriching quantity (μ g/ basin) | 221.2±11.6 | 157.8±24.1 | 0.402 |
| ? | Root Cu enriching quantity (μ g/ basin) | 201.7±23.6 | 118.1±8.3 | 0.709 |
| Sin | Overground part Cu enriching quantity (μ g/ basin) | 156.6±12.5 | 77.0±7.4 | 1.034 |
| ? | Root Cu enriching quantity (μ g/ basin) | 261.8±7.5 | 187.3±25.3 | 0.398 |
| Pur | Overground part Cu enriching quantity (μ g/ basin) | 246.3±9.4 | 153.1±6.7 | 0.609 |
| ? | Root Cu enriching quantity (μ g/ basin) | 220.5±2.6 | 162.7±10.2 | 0.355 |
In embodiment 9 heavy-metal contaminated soils, root nodule bacterium W33 promotes the effect of alfalfa growing and dross
Gather the soil of heavy metal cuprum polluted (total copper content be 850mg/kg), air-dry grinding, the plastic tub of packing into alms bowl, every basin 2kg, adding water, to make water content be 60% of field capacity, keeps 2d, every basin is sown into 20 of alfalfa seeds.Inoculation embodiment 6 gained take the biological restoration microbial inoculum 10ml that W33 bacterial strain (CCTCC NO:M 2012357) be effective constituent, contrast and connect the equivalent sterilized water.During plant strain growth, add distilled water with weighting method every day, and to keep soil humidity be field capacity 60%.What growth inoculated embodiment 6 gained after 30d take biological restoration microbial inoculum 10ml that the W33 bacterial strain is effective constituent once.Sowing 60d results along native face clip plant overground part, wash out root system simultaneously, the root nodule on the counting root system of plant.Root system is completed under 105 ℃, 70 ℃ of oven dry, the dry weight of weighing root and overground part.As shown in Table 5, bacterial strain W33 can obviously promote the growth of clover in heavy-metal contaminated soil, with connect contrasting of sterilizing microbial inoculum and compare, inoculation be take the biological restoration microbial inoculum that the W33 bacterial strain is effective constituent and is made the plant height of clover significantly increase by 1.7 times, root is heavy increases respectively 2.7 and 2.4 times with the overground part dry weight, reaches utmost point conspicuous level.And, W33 can be on clover dross, visible clover is effective symbiosis host of root nodule bacterium W33.
The growgh promoting effects of root nodule bacterium W33 to clover in table 5 heavy-metal contaminated soil
| Project | Connect bacterium | Contrast |
| Plant height (cm) | 20.5±5.2 | 10.0±1.1 |
| Root heavy (g/ basin) | 0.84±0.32 | 0.23±0.05 |
| Overground part dry weight (g/ basin) | 1.46±0.49 | 0.43±0.06 |
| The dross amount (grain /) | 4.7±2.1 | 0.7±0.6 |
The root nodule bacterium W33 Promoting plant growth effect of anti-heavy metal on embodiment 10 tungsten ore tailings abandoned lands
By take after biological restoration microbial inoculum 2kg that the W33 bacterial strain is effective constituent and 4kg tungsten ore tailings, 200g weeping love grass seed mixing of embodiment 7 gained, be seeded in tungsten ore abandoned land community, cell size is 1m * 1m, repeats 3 times.Contrast inoculation sterilizing W33 biological restoration microbial inoculum.Establish altogether 6 communities.Again inoculate the biological restoration microbial inoculum that the W33 bacterial strain is effective constituent of take of embodiment 7 gained after January wait the growth of emerging, spread fertilizer over the fields after 2kg microbial inoculum and 8kg soil are mixed thoroughly.Regularly water, weeding.Harvesting after 3 months, measure plant height, dries to weight, measures biomass.The results are shown in Table 6.Bacterial strain W33 can promote weeping love grass plant height and dry weight significantly to increase, and improves 30% and 43% than contrast respectively.
The growgh promoting effects of root nodule bacterium W33 to weeping love grass on table 6 tungsten ore abandoned land
On embodiment 11 rare earth tailings abandoned lands, root nodule bacterium W33 promotes the weeping love grass growth effect
By take after biological restoration microbial inoculum 2kg that the W33 bacterial strain is effective constituent and 4kg rare earth tailings, 200g weeping love grass seed mixing of embodiment 7 gained, be seeded in the rare earth community, cell size is 1m * 1m.Contrast inoculation sterilizing W33 biological restoration microbial inoculum is again inoculated the biological restoration microbial inoculum that the W33 bacterial strain is effective constituent of take of embodiment 7 gained after the growth January of emerging, and after 2kg microbial inoculum and 8kg soil are mixed thoroughly, spreads fertilizer over the fields.Extract plant after 3 months, measure root length and plant height.Measure now fresh weight and be designated as W1, then clean and remove grit, slightly blot rear gravimetry with thieving paper and be designated as W2.According to formula W, fix the sand=W1-W2 calculates the amount of fixing the sand.The results are shown in Table 7.Bacterial strain W33 can promote weeping love grass root length and plant height significantly to increase, and improves 19% and 46% than contrast respectively.The increase of plant dry weight is not remarkable.Bacterial strain W33(CCTCC NO:M 2012357) inoculation makes the root system of plant prosperity, and the amount of fixing the sand significantly increases by 98%, is conducive to the soil conservation of rare-earth tailing abandoned land.
The growgh promoting effects of root nodule bacterium W33 to weeping love grass on table 7 rare earth abandoned land
Claims (10)
1. a root nodule bacterium W33, Classification And Nomenclature is Rhizobium sp.W33, is preserved in Chinese Typical Representative culture collection center, and preservation date is on September 18th, 2012, and culture presevation number is CCTCC NO:M 2012357.
2. a biological restoration preparation that contains the root nodule bacterium W33 that preserving number claimed in claim 1 is CCTCC NO:M 2012357.
3. biological restoration preparation according to claim 2, is characterized in that this biological restoration preparation is liquid preparation, and the W33 bacterial strain living bacteria count that wherein to contain preserving number be CCTCC NO:M 2012357 is more than 1,000,000,000/milliliter.
4. biological restoration preparation according to claim 3, is characterized in that described liquid preparation is to obtain by the conventional liquid submerged fermentation method described root nodule bacterium W33 that ferments.
5. biological restoration preparation according to claim 2, is characterized in that this biological restoration preparation is solid preparation, the W33 bacterial strain living bacteria count that wherein to contain preserving number be CCTCC NO:M 2012357 be 200,000,000/more than gram.
6. biological restoration preparation according to claim 5, is characterized in that described solid preparation is granule or the pulvis that uses root nodule bacterium W33 fermented liquid that the peat composed of rotten mosses, vermiculite or other carrier adsorption preserving numbers are CCTCC NO:M 2012357 to be mixed with.
7. the application of root nodule bacterium W33 in mine tailing district phytoremediation that preserving number claimed in claim 1 is CCTCC NO:M 2012357.
8. the application of root nodule bacterium W33 in the phytoremediation of heavy-metal contaminated soil that preserving number claimed in claim 1 is CCTCC NO:M 2012357.
9. the application of biological restoration preparation in the phytoremediation of mine tailing district or heavy-metal contaminated soil of root nodule bacterium W33 claimed in claim 2.
10. the biological restoration preparation of the root nodule bacterium W33 of containing claimed in claim 2 is used for the plant restoration method of heavy metal pollution of soil, it is characterized in that: in the moistening soil containing heavy metal or mine tailing district, sow plant seed, inoculate the described biological restoration preparation containing root nodule bacterium W33, every mu of inoculation 10
8The biological restoration preparation 4-6kg of individual bacterium/mL, minute 1-2 inoculation.
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| CN105418204A (en) * | 2014-09-02 | 2016-03-23 | 南京农业大学 | New applications of Rhizobium W33 strain |
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