CN102921714B - A kind of exotrophic mycorrhiza planting technology for high concentration copper polluted soil - Google Patents
A kind of exotrophic mycorrhiza planting technology for high concentration copper polluted soil Download PDFInfo
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- CN102921714B CN102921714B CN201210429188.2A CN201210429188A CN102921714B CN 102921714 B CN102921714 B CN 102921714B CN 201210429188 A CN201210429188 A CN 201210429188A CN 102921714 B CN102921714 B CN 102921714B
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Abstract
The present invention relates to field of environment protection, is a kind of exotrophic mycorrhiza planting technology for high concentration copper polluted soil, is particularly useful for phytoremediation plant growth being had to the copper polluted soil of obviously poisoning.High concentration copper polluted soil has higher murder by poisoning to plant due to excess copper, and therefore, plant often poor growth maybe can not survive, and adopts the technology of extensive earthing often to need a large amount of foreign aid's mulching soils and Engineering operation cost costly.This technological invention is the method for a kind of bag balled transplanting, places a small amount of uncontamination soil, and in root bag, plants the plant of exotrophic mycorrhiza, be transplanted in contaminated soil by this root bag (containing soil and plant) in root bag.Root bag can reduce the diffusion of the Cu in ambient contamination soil to root system of plant, and simultaneously exotrophic mycorrhiza mycelia can strengthen the absorption of plant to Pi and moisture, thus significantly improves biomass and the survival ability of plant.This technological invention is particularly useful for revegetation plant being had to the high concentration copper polluted soil of obviously poisoning.
Description
Technical field
The present invention relates to field of environment protection, be a kind of heavy metal pollution carry out the novel plant implantation methods of revegetation, be particularly useful for phytoremediation plant growth being had to the high concentration copper polluted soil of obviously poisoning.
Background technology
At present, the important component part of the ecological environment that soil is depended on for existence as the mankind, is on the rise by heavy metal pollution.Hg about 1.5 ten thousand tons is discharged in the whole world every year on average, Cu340 ten thousand tons, Pb500 ten thousand tons, Mn1500 ten thousand tons, Ni100 ten thousand tons.China's heavy metal pollution of soil is also quite serious, and be discharged into toxic heavy metal in environment every year up to millions of tons, wherein Pb is 34.6 ten thousand tons, and Cd is 3.9 ten thousand tons (Nriagu and Pacyna, 1988).
Current, China's regional agriculture environment worsens with agricultural product heavy metal pollution phenomenon very serious, particularly in some developed areas.At present, the whole nation suffers the cultivated area polluted in various degree close to 2000 × 10
4hm
2, account for 1/5 of cultivated area.According to statistics, China's industrial triwaste pollution cultivated area 266.7 × 10 in 1980
4hm
2, within 1988, be increased to 666.7 × 10
4hm
2, within 1992, be increased to 1000 × 10
4hm
2(from, 2000).Ministry of Agriculture's investigation shows: China's irrigating region area about 140 × 10
4hm
2, accounted for by the land area of heavy metal pollution and pollute 64.8% of the gross area, wherein slight pollution accounts for 46.7%, and intermediate pollution accounts for 9.7%, and severe contamination area accounts for 8.4%, wherein maximum with the contaminated area of Hg and Cd.The whole nation about has 1.3 × 10 at present
4hm
2plough and be subject to the pollution of Cd, relate to 25 areas (Chen Huaiman, 1996) of 11 provinces and cities.It is reported, Cd content severe overweight in the rice that current China irrigating region has 11 places to produce.Such as, the arable land that county of Jiangxi Province reaches 44% is polluted, and forms 670hm
2" cadmium rice " district (Li Lujiu, 2004).The grain drop in production that China causes because of heavy metal pollution is more than 1000 × 10
4t/a, contaminated by heavy metals grain about 1200 × 10
4t/a, adds up to economic loss about 20,000,000,000 yuan/a (Wei Chaoyang, 2001).Therefore, the regulation in heavy metal pollution soil and improvement, become the top priority that China improves the ecological environment, ensures people's life health.
Heavy metal pollution of soil is administered and is mainly adopted the engineering measure such as physics, chemistry, but these methods engineering amounts greatly, complicated operation, somewhat expensive, some can't fundamentally solving practical problems, easily causes secondary pollution.People find in research afterwards, and phytoremediation has efficiently, it is low, easy to operate to invest, do not cause the advantages such as secondary pollution, provide an important reparation and governance way for fundamentally solving heavy metal pollution of soil.At the beginning of the eighties in last century, Bradley (1981) etc. find when investigating Britain's mining-area plant, and in the mining area that metal especially content of beary metal is very high, it is good that mycorrhizal plants grows than non-mycorrhizal plants.After article is delivered on " NATURE ", open the frontier of mycorrhizal plants preventing from heavy metal research, also pulled open the prelude utilizing mycorrhizal plants to repair with carrying out heavy metal pollution.Many scientists, especially ecology and physiologist, under heavy metal contaminated environment, the metabolism and growth reaction of mycorrhizal plants is studied, and inquires under excessive heavy metal stress environment, mycorhiza is to the possible protective effect of host plant and mechanism (Denny, 1995; Diaz, 1996; Hartley, 1997), thus provide fundamental basis for application mycorrhizal plants carries out soil remediation.
Up to the present, people apply that a lot of method fixes, enrichment, the excessive heavy metal copper of transfer, cadmium, and wherein apply the method that the fixing heavy metal of mycorrhizal plants accumulation is a practicality and great potential.Such as, Huang Yi (2001) points out that exotrophic mycorrhiza can increase the accumulation of host plant to copper, and the content of its aerial part declines, and under ground portion content increases; Cu process exotrophic mycorrhiza foot end Cu content is 2.6 times of sterilized root plant.Jentschke (1999) research finds that the Cd of the Norway pine energy enduring high-concentration of inoculation Paxillus involutus pollutes, and points out that mycorhiza has larger absorption and fixation to Cd.Therefore, mycorrhizal plants is utilized to have important using value with repairing the heavy metal pollution such as copper, cadmium.
The murder by poisoning that high concentration heavy-metal contaminated soil is higher because heavy metal on plants has, therefore, plant often poor growth maybe can not survive, and adopts the technology of extensive earthing often to need a large amount of foreign aid's mulching soils and Engineering operation cost costly.This technological invention is the method by a kind of bag balled transplanting, places a small amount of uncontamination soil, and in root bag, plants the plant of exotrophic mycorrhiza, be transplanted in contaminated soil by this root bag (containing soil and plant) in root bag.Root bag can reduce the diffusion of the Cu in ambient contamination soil to root system of plant, and simultaneously exotrophic mycorrhiza mycelia can strengthen the absorption of plant to Pi and moisture, thus improves biomass and the survival ability of plant.This technological invention is particularly useful for revegetation plant being had to the high concentration copper polluted soil of obviously poisoning.
Summary of the invention
The invention provides the implantation methods of a kind of novel exotrophic mycorrhiza plant at high concentration copper polluted soil.
Beneficial effect
The exotrophic mycorrhiza plant cover cultivation methods that the present invention adopts, in root bag, the planting and inoculating exotrophic mycorrhiza pine tree of purple wax mushroom (Laccariaamethystea (bull.Ex Gray) Murr), is embedded in root bag in the Cu contaminated soil of variable concentrations.Potted plant experiment shows, the plant of plantation in root bag, and biomass remains on a higher level and by the impact of the outer contaminated soil copper toxicity of root bag.Although the velamen of plant is limited in root bag, but, mycorhiza can pass root bag thus expand the nutrient absorption scope of plant, the Mycorrhizal seedling overground part of root bag process and the Pi total amount of Root Absorption are significantly higher than the Pi amount of control group Mycorrhizal seedling Root Absorption, and, mycorhiza through root bag can also activate and promote that Cu is to the transhipment of plant root, although the content of Cu in plant root, stem and needle all improves with the increase of extraneous Cu concentration, Cu accumulation is significantly lower than the Cu content in extraneous soil.Illustrate, root bag can reduce the diffusion of the Cu in ambient contamination soil to root system of plant, and simultaneously exotrophic mycorrhiza mycelia can strengthen the absorption of plant to Pi and moisture, thus significantly improves the survival ability of plant on high concentration copper polluted soil.
Accompanying drawing explanation
Fig. 1 experimental provision sketch
Fig. 2 phytomass
The total Cu accumulation of Fig. 3
Fig. 4 overground part phosphorus content
Fig. 5 root system phosphorus content
Detailed description of the invention
The pure culture of 1 exotrophic mycorrhiza
Purple wax mushroom (Laccaria amethystea (bull.ExGray) Murr) (LA) of ectotrophic mycorrhiza is by this Laboratories Accession.
(1) be down flat plate MMN culture medium is configured, mix, after high pressure steam sterilization, be down flat plate.MMN culture medium prescription is: maltose 3g, glucose 10g, (NH
4) HPO
40.25g, KH
2pO
40.5g, MgSO
47H
2o0.15g, CaCl
20.05g, FeCl
36H2O0.02g, NaCl0.025g, VB10.1mg, agar 13g.
(2) cutting single bacterium block, to choose the MMN being vaccinated with LA dull and stereotyped, cuts out the fritter of 5mm × 5mm at colony edge, and back-off is with new MMN flat board, and sealing, 25 DEG C of cultivations 1 month are stand-by.
The acquisition of 2 Mycorrhizal plants
(1) nursery Korean pine (Pinus koraiensis S.) seed with 30% H
2o
2sterilize clean with deionized water rinsing after 30 minutes, be seeded in afterwards in vermiculite, 25 DEG C of 8 weeks of cultivation are stand-by.
(2) Mycorrhizal is placed a filter paper and is spread the volcanic ash of 1cm thickness bottom basin alms bowl: rural area soil=1: 1 mixed-matrix.Stromal surface places the LA bacterium colony that grown about month.The pine tree seedling that grown about 8 weeks wipes out part side root, and contacted with bacterium colony by its root, surrounding fills up mixed-matrix, microscopy after Mycorrhizal three months.
The detection of 3 Mycorrhizal plants
Stereo microscope monitor station is put a clean dull and stereotyped the inside and hold 15ml deionized water, the Korean Pine Seedlings after Mycorrhizal three months is placed in flat board, the side root that statistics mycorhiza infects and total side radical order, and infection rate is side radical/total side radical that mycorhiza infects.Choose the test that Mycorrhizal seedling that infection rate is greater than 90% carries out below.
The preparation of 4Cu contaminated soil
The CuSO of artificial interpolation 0,1.25,2.5,5.010.0mg/ml
4solution, in volcanic ash, makes the Cu concentration in soil be 0,375,750,1500,3000mg/kg, mixes.Soil 121 DEG C of sterilizing 120min of 5 kinds of Cu process, are dispensed in five plastic tub alms bowls, every basin 550g after cooling.
5 bagging apparatus
The bottom of 50ml centrifuge tube amputates, the nylon wire in 2 layers of 40 μm of aperture on whorl side pad, and therefore the root of plant cannot pass nylon wire, and mycorhiza can pass.Often transplant two strain Mycorrhizal seedling (infection rate is close to 100%) in pipe centrifuge tube, surrounding fills the free of contamination andosol of 40g.3 pipe nylon wire centrifuge tubes placed respectively by 5 basins Cu contaminated soil basin alms bowl in various degree.
Identical with nylon wire centrifuge tube, other 5 pipe centrifuge tubes cut away bottom equally, and whorl side does not pad nylon wire, tightens with lid.Often pipe centrifuge tube transplants 2 strain Mycorrhizal seedling, fills the free of contamination andosol of 40g around.1 pipe control group centrifuge tube placed respectively by 5 basins Cu contaminated soil basin alms bowl in various degree.
The results of 6 plants and metal assay
The 120th day results plant after plant transplantation, plant washed with de-ionized water root, is divided into overground part and root system afterwards.80。C is dried 72 hours and is weighed, and grinding sieves makes particle be less than 1mm.
The assay method of Cu content in sample: the plant sample agate mortar of oven dry grinds mixing, take plant dry sample 0.2000g to be placed in disappear and to boil pipe, disappear with HNO3-HClO4 (V: V=87: 13) mixed liquor and boil (Zhao etc., 1994), analytikjena-nova400 flame atomic absorption spectrophotometer (AAS) measures Cu content.In analytic process, the standard specimen NIST (SRM2709) firmly believed using American National Standard and technological associations is used for disappearing as a part of QA/QC and boils.Be provided with suitable blank in experiment also and repeat with detection accuracy and accuracy.The heavy metal recovery rate of standard soil sample (NIST SRM2709) will be controlled in 90 ± 5% scopes.Analyzing agents useful for same all adopts top grade pure, and water used is deionized water.
Fig. 2 shows in Cu contaminated soil, the dry weight of plant plant above ground portion and root system under variable concentrations Cu process of Ectomycorrhizal Inoculation bacterium.As can be seen from Figure, with the plant seedling that nylon wire cuts off under the Cu process of five kinds of concentration, the upperground part biomass, all higher than control group, is respectively 1.97,1.94,1.95,2.24,2.22 times of contrast; The biomass of the root system of Mycorrhizal seedling also higher than control group, compared with the control, weight of root system respectively than contrast high by 38.4%, 17.9%, 43.4%, 47.9%, 14.3%, the growth of visible exotrophic mycorrhiza mycelia to plant has facilitation.
From Fig. 3 and table 1, the Cu total amount of the Mycorrhizal seedling Root Absorption of root bag process is significantly higher than the Cu of control group Mycorrhizal seedling Root Absorption.This is mainly because mycelia through nylon wire, can expand the absorption region of Cu, more obvious in high concentration Cu contaminated soil.
In table 1 varying level copper polluted soil, root bag is on the impact (ugpot of total Cu amount that Mycorrhizal pine tree seedling absorbs
-1)
Numerical value is three mean value ± standard deviations repeated.Numerical value letter below (a, b, c) as identical, then represents that two values do not have difference in P<0.05 level; Letter is different, then there were significant differences in P<0.05 level to represent two values.
7 connect the impact of bacterium process on plant absorption P
In plant, phosphorus content is with reference to Shi Rui and the method with Bao Shidan (1980).Get plant dry sample and be about 0.1g, add the HClO of 1ml50%
4h dense with 5mL
2sO
4, spend the night after shaking up, then boil in stove to disappear in disappearing and boil, boil liquid become colorless until disappear, be settled to 25mL by deionized water after cooling, shake up standing.Get 1mL to disappear and boil liquid, add 10mL deionized water, 2 2,4-DNP nitrite ions, use 4mo1L
-1naOH be adjusted to and just occur yellow till, then use 2molL
-1h
2sO
4be adjusted to colourless.Add the anti-reagent of 2.5mL molybdenum antimony (ammonium molybdate+potassium antimony tartrate ten ascorbic acid), be settled to 25mL, shake up by deionized water, reaction 30min, measures light absorption value, boils liquid as blank not add disappearing of sample under 700nm wavelength.To analyze pure KH2P04 as standard sample production standard curve.
In table 2 varying level copper polluted soil, root bag absorbs the impact of Pi on Mycorrhizal pine tree seedling
Numerical value letter below (a, b, c) as identical, then represents that two values do not have difference in P<0.05 level; Letter is different, then there were significant differences in P<0.05 level to represent two values.
The Pi being significantly higher than control group Mycorrhizal seedling Root Absorption by the Pi total amount of the Mycorrhizal seedling overground part in the visible root bag of Fig. 4 Fig. 5 and table 2 and Root Absorption measures.This is mainly because mycelia through nylon wire, can expand the absorption region of Pi.Ectotrophic mycorrhiza can promote that plant is to the absorption of Pi, and this may be the major reason of its Promoting plant growth.
The present embodiment shows: root bag can reduce the diffusion of the Cu in ambient contamination soil to root system of plant, exotrophic mycorrhiza mycelia can absorb Pi and the moisture of outside contamination soil through nylon wire, the growth of significant promotion plant above ground portion and root system, contributes to the field planting of the plant on heavy metal pollution ground.
Claims (3)
1. the exotrophic mycorrhiza plant cover cultivation methods for high concentration copper polluted soil, it is characterized in that adopting the plantation of root bag, described bag is that the nylon wire in 25 to 40 μm of apertures is made, described bag is built with the uncontamination soil to plant growth nonhazardous, the Mycorrhizal plant of the external bacterium inoculation of plantation in soil, the root system of described plant cannot pass described nylon wire, and described Applying Ectomycorrhizal Fungi mycelia can pass described nylon wire, wherein, described plant is can form the pine tree of symbiosis, robur or willow with Applying Ectomycorrhizal Fungi, and described external bacterium is purple wax mushroom; Described bag cave is planted and is transplanted in the soil of high concentration Cu-W ore deposit.
2. exotrophic mycorrhiza plant cover cultivation methods according to claim 1, is characterized in that, described exotrophic mycorrhiza mycelia can absorb the copper in extraneous soil through described bag, significantly improve the copper content of described plant shoot and root system.
3. exotrophic mycorrhiza plant cover cultivation methods according to claim 1, it is characterized in that, described exotrophic mycorrhiza mycelia can absorb phosphorus in extraneous soil and moisture through described bag, significantly improve the phosphorus content of described plant shoot and root system, promote the growth of plant shoot and root system, contribute to the field planting of plant on high concentration Cu-W ore deposit ground.
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| CN103861865B (en) * | 2014-03-04 | 2016-03-02 | 张菁菁 | A kind of method utilizing natural void soil conservation fixing device repairing polluted soil |
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| CN107616096A (en) * | 2017-11-08 | 2018-01-23 | 南京林业大学 | A kind of aseptic pine seedlings cultural method |
| CN108856264A (en) * | 2018-06-28 | 2018-11-23 | 上海环境工程设计研究院有限公司 | A kind of Compound Heavy Metals phytoremediation for soil method |
| CN109127686A (en) * | 2018-07-31 | 2019-01-04 | 湖南省园林建设有限公司 | Heavy-metal contaminated soil mycorrhizal plants root bag implantation methods |
| CN110313390A (en) * | 2019-08-02 | 2019-10-11 | 金埔园林股份有限公司 | Utilize the method for Applying Ectomycorrhizal Fungi auxiliary black pine improvement heavy-metal contaminated soil |
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