CN105750212A - Method for screening varieties of heavy metal hyperaccumulation plants - Google Patents
Method for screening varieties of heavy metal hyperaccumulation plants Download PDFInfo
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Abstract
The invention provides a method for screening the varieties of heavy metal hyperaccumulation plants and relates to the technical fields of agriculture and soil treatment.The method includes: setting screening standard concentration, selecting the varieties of primary-screening plants, and screening the varieties of the heavy metal hyperaccumulation plants.The method is low in operation workload, small in required area, low in cost, short in selection cycle, good in result stability, high in practicality, and suitable for being widely popularized.
Description
Technical field
The present invention relates to agricultural technology field and soil remediation technical field, in particular to the screening technique of the kind of a kind of super enrichment heavy metal plant.
Background technology
Soil is one of important resource of human society, is developing agricultural, industry and the material base carrying out urban modernization construction.But, owing to the strong of mankind's activity such as mining and smelting, industry and traffic emission, garbage stacking, pesticide and fertilizer application affect, soil pollution is day by day serious.Especially the exploitation of mineral resources and smelting, while providing grand strategy resource for country, also result in the serious consequences such as environmental pollution, geological disaster, ecological disruption.
And in numerous remediation contaminated soil methods, compared with traditional chemically and physically Treatment process, phytoremediation (phytoremediation) by its economy, environmental friendliness, the feature such as soil productivity, non-secondary pollution can be kept, receive the extensive concern of academia.Phytoremediation technology includes utilizing plant ultraproduct tired or accumulating the Phytoextraction reparation of sexual function, utilize root system of plant control contamination and recover the Phytostabilization of ecological functions, utilize the phytovolatilization reparation of Plant Transformation function.At present, generally believe, Phytoextraction reparation is applicable to the agricultural land soil that heavy metal pollution degree is not as high, through plantation several seasons or several years super enrichment heavy metal plant, by the heavy metal accumulation in soil to aboveground vegetation part, then process further after adopting conventional agriculture method results, finally make heavy metal-polluted soil concentration drop to below standard of soil environment quality, make cereal crops to keep the safety in production.Plant stability technology is then more suitable for the ecological recovery in the high heavy metal pollution place that industry and mining cause, to reduce the diffusion mobility of the heavy metal element that rainwash erosion, wind erosion etc. cause.
In prior art, screen the kind of super enrichment heavy metal plant, first having to can growing plants at highly polluted regional observation, then it is sampled collecting to the plant having enrichment potentiality, the mode that liquid culture or earth culture are supported all is adopted to cultivate, then the content of heavy metal in soil, plant shoot and root is checked, it may be judged whether for the kind of super enrichment heavy metal plant.This kind of method workload is big, and demand area is big, and cost is high, and the cycle is long, and result stability is also poor.
Summary of the invention
It is an object of the invention to provide the screening technique of the kind of a kind of super enrichment heavy metal plant, it is little that this screening technique has workload, and demand area is little, and cost is low, the advantage that selection cycle is short.
This invention address that it technical problem is that to realize by the following technical solutions:
The screening technique of the kind of a kind of super enrichment heavy metal plant, comprises the following steps:
Set screening criteria concentration: be respectively put into by many seeds of the plant through pretreatment in multiple Heavy metal solution that concentration is different, carry out germinateing and cultivate, the concentration choosing the Heavy metal solution at the seed place of the plant that seedling-erecting ratio is 50% is screening criteria concentration, seedling-erecting ratio=(B/C) 100%, B is upright Seedling number, and C is germination number；
Choose the kind of primary dcreening operation plant: select multiple kinds of plant, seed by each kind through pretreatment, put in the heavy metal screening solution with screening criteria concentration, carry out germinateing and cultivate, calculate the seedling-erecting ratio of the seed of each kind, select seedling-erecting ratio high front 10～20% the kind that kind is primary dcreening operation plant corresponding to seed；And
The kind of screening super enrichment heavy metal plant: the seed of the kind of each primary dcreening operation plant through pretreatment is seeded in the many parts of soil that heavy metal concentration is different respectively, and carry out field management, until it is grown to plant, measure the content of beary metal of the root of the content of beary metal of every part of soil and plant corresponding to every part of soil and the content of beary metal of plant shoot, when plant is survived, and the content of beary metal of its plant shoot is higher than the content of beary metal of root, and the substance withdrawl syndrome of its plant shoot is more than the substance withdrawl syndrome of root, and the content of beary metal before the soil remediation at its place more than administer after content of beary metal time, determine the kind that the kind belonging to plant is super enrichment heavy metal plant.
Relative to prior art, the present invention includes following beneficial effect: the present invention, by arranging multiple test concentrations, then carries out the selection of science, it is determined that the base concentration that plant can adapt to, obtains normal concentration according to half lethal concentration；Then, utilizing step-sizing method, the bud phase sensitive by plant heavy metal carries out Preliminary screening；Screen further again through potted plant mode, it is determined that the absorbability of plant heavy metal, determine the kind of super enrichment heavy metal plant by detecting content of beary metal.Wherein, introduce seedling-erecting ratio concept, utilize the plant heavy metal highstrung bud phase to carry out Preliminary screening, eliminate the material that heavy metal is sensitive, significantly decrease workload, improve work efficiency, by calculating seedling-erecting ratio rapidly, screen accurately.Since, when the seed of plant germinates under the coercing of heavy metal environment, in fact it could happen that: (1) seed is not sprouted；(2) after seed germination, in several days subsequently, the amount of heavy metal owing to absorbing is more, and the growth of the root of seed is suppressed, root root short, not long or root blackening, and it can not uprightly flat crouch；(3) seed can be sprouted, and root growth is normal, and stem and leaf also is able to normal growth, and this kind is just for kind that super enrichment ability is stronger.
Being additionally, since the plant germination phase is generally several days, it is possible to repeatedly screen at short notice, the biological volume of plant sprout phase is also less, and area occupied is little, and indoor just can be screened, also not by the impact of the natural causes such as rainwater in experimentation, it is simple to manual adjustment.The method is simple to operate, and less input for cost, it makes use of multiple plant variety, multiple heavy metal concentration is tested, and scope of experiment is big, and data are reliable, and result stability is better, and selection cycle is also short.
Detailed description of the invention
For making the purpose of the embodiment of the present invention, technical scheme and advantage clearly, the technical scheme in the embodiment of the present invention will be clearly and completely described below.Unreceipted actual conditions person in embodiment, conventionally the condition of condition or manufacturer's suggestion carries out.Agents useful for same or the unreceipted production firm person of instrument, be and can pass through the commercially available conventional products bought and obtain.
Below the screening technique of the kind of the super enrichment heavy metal plant of the embodiment of the present invention is specifically described.
The screening technique of the kind of super enrichment heavy metal plant, comprises the following steps:
Set screening criteria concentration: be respectively put into by many seeds of the plant through pretreatment in multiple Heavy metal solution that concentration is different, carry out germinateing and cultivate, the concentration choosing the Heavy metal solution at the seed place of the plant that seedling-erecting ratio is 50% is screening criteria concentration, seedling-erecting ratio=(B/C) 100%, B is upright Seedling number, and C is germination number；
Choose the kind of primary dcreening operation plant: select multiple kinds of plant, seed by each kind through pretreatment, put in the heavy metal screening solution with screening criteria concentration, carry out germinateing and cultivate, calculate the seedling-erecting ratio of the seed of each kind, select seedling-erecting ratio high front 10～20% the kind that kind is primary dcreening operation plant corresponding to seed；And
The kind of screening super enrichment heavy metal plant: the seed of the kind of each primary dcreening operation plant through pretreatment is seeded in the many parts of soil that heavy metal concentration is different respectively, and carry out field management, until it is grown to plant, measure the content of beary metal of the root of the content of beary metal of every part of soil and plant corresponding to every part of soil and the content of beary metal of plant shoot, when plant is survived, and the content of beary metal of its plant shoot is higher than the content of beary metal of root, and the substance withdrawl syndrome of its plant shoot is more than the substance withdrawl syndrome of root, and the content of beary metal before the soil remediation at its place more than administer after content of beary metal time, determine the kind that the kind belonging to plant is super enrichment heavy metal plant.
Wherein, upright Seedling to be cotyledon green can adhere to without yellow, stem without lodging sign, root and can absorb water, can the Seedling of normal growth.
In the present invention, heavy metal solution is at least one in Caddy (Cleary) 2/5 hydrate soln, cadmium nitrate solution.Preferred Caddy (Cleary) 2/5 hydrate soln.Certainly, this kind of setting is not intended as restriction, it practice, heavy metal solution should set according to actual heavy metal pollution, chooses experimental solutions, simulating pollution environment.Can be selected for that Herba pteridis vittatae carries out administering, lead contamination can be selected for roundleaf Thalaspi arvense L. and administers as arsenic pollutes.
If heavy metal solution selects cadmium-containing solution, then plant is preferably Brassica campestris L.Because nursery is it is experimentally confirmed that Brassica campestris L is optimal plant for the reparation of cadmium pollution, it has some advantage following:
(1) cadmium is had superpower absorbability by Brassica campestris L, is one of plant that the level of accumulation heavy metal in the world is the highest, and the accumulation capability of cadmium is reached 2000ppm by Brassica campestris L, and the accumulation capability of zinc is reached 1000ppm, and in general, the purification cycle of Brassica campestris L is 3～5 years；
(2) Oilseed rape biomass is big, be prone to harvesting, and its climatic adaptation ability is also relatively strong, and Brassica campestris L belongs to unique large oil crop in winter, soil out of question with Oryza sativa L., will not reduce the income of peasant, and south rice field major part is left unused winter, light warm water is sufficient, is suitable for rapeseed cultivation；
(3) if the Semen Allii Tuberosi planted out is without cadmium, then the oil squeezed out may be used for eating；If the Semen Allii Tuberosi planted out is containing cadmium, then the oil squeezed out may be used for power plant, it is to avoid throws aside and causes the secondary pollution of cadmium；
(4) china rape refers generally to cabbage type rape, belongs to same genus with Indian mustard, and two species genes have high homology, it is easy to filter out and select the kind of super enrichment heavy metal plant from cabbage type rape.
Certainly, choosing of Brassica campestris L is not intended as restriction, and the selection of plant should be determined according to actual heavy metal pollution.If cadmium pollution, then except Brassica campestris L, plant can also be chosen as Thlaspi caerulescens, Sedum alfredii Hance, pteridophyte Asplenium trchomanes L., wild Amaranthus mangostanus L. etc., simply Sedum alfredii Hance, pteridophyte Asplenium trchomanes L., wild Amaranthus mangostanus L. Biomass too little, plant is shorter, results are difficult to mechanization, and the absolute magnitude of cadmium is very limited.And the Biomass of Indian mustard is less than normal, more weak at the adaptive capacity of China, administer limited potential for cadmium pollution.
For the ease of describing, in embodiments of the invention, heavy metal solution is disposed as cadmium solution, and plant all selects Brassica campestris L.
When setting screening criteria concentration, Comprehensive Traits can be selected excellent or promote mainly 1～2 variety seeds of kind, then Concentraton gradient reasonable in design, as 10mg/kg, 20mg/kg, 40mg/kg, 60mg/kg, 80mg/kg, 100mg/kg, 120mg/kg seven cadmium solutions (Heavy metal solution) process and a control treatment, each process at least two repeats.It is of course also possible to be set to other Concentraton gradient, such as 30mg/kg, 50mg/kg, 70mg/kg.
When setting screening criteria concentration, plant seed is respectively put in multiple Heavy metal solution that concentration is different, seed in each culture dish carries out the process of the Heavy metal solution of a concentration, process and a control treatment if any seven cadmium solutions (Heavy metal solution), then need with eight culture dishs, the corresponding process of each culture dish；If each process two repetition, then need to add eight culture dishs again, corresponding two culture dishs of each process.Each culture dish loads a quantity of seeds, such as 50,43,66.In the pretreatment setting screening criteria concentration step, including the seed in each culture dish all being carried out sterilizing.Can utilizing the medicining liquid dipping seed 8～12min of 4～6g/kg, so, it is possible to prevent later stage seed from going mouldy, this operation simultaneously also has urges the effect planted；Then, utilization processes water and cleans seed, washes the disinfectant solution of the surface of the seed attachment off, it is to avoid affect later experiments.Wherein, disinfectant solution is at least one in sodium hypochlorite, potassium permanganate solution, mercuric chloride solution, and processing water is at least one in the deionized water after pure water, ultra-pure water, sterilizing, selects these water to process, it is possible to be prevented effectively from introducing antibacterial.
In addition, when to seed sterilizing, in order to sterilize thoroughly, it is also possible to before using disinfectant solution, first with the alcohol-pickled seed 4～6min of 700～800g/kg, then ethanol is poured out, add disinfectant solution to process, after disinfectant solution processes, pour out disinfectant solution, then clean seed 2～4 times with pure water continuously, then clean one time with ultra-pure water or sterilized water for injection.
After setting the pretreatment of screening criteria concentration step, then seed is moved on in culture dish, repetition and comparison are set simultaneously, in culture dish, then add 3～5ml cadmium solution (Heavy metal solution) prepared, carry out labelling.Shifting at seed is let slip in journey, it is preferable that place mat 2～3 metafiltration paper under seed, and so, after cadmium solution drips, seed can better also absorption cadmium solution evenly.And when preparing cadmium solution, it is preferable that utilize Caddy (Cleary) 2/5 hydrate and process water to prepare.
Then, all of culture dish is placed in temperature 22～25 DEG C, periodicity of illumination be bright phase 15～17h, dark phase 7～9h environment in carry out germinate cultivate.Until the 6th～8 day, adding up seedling-erecting ratio.Needing in seedling-erecting ratio to calculate upright Seedling number, i.e. the quantity of upright Seedling, upright Seedling refers to that cotyledon green is without yellow, and stem is without lodging sign, and root can be adsorbed on filter paper and can absorb water, can the Seedling of normal growth.Certainly, if not being lined with filter paper in culture dish, then can observe whether its root can adhere to and can absorb water.Additionally, when adding up seedling-erecting ratio, it is also possible to add up germination percentage simultaneously, so, it is possible to ensure that experimental data is more accurately, reliably.In general, the normal concentration of Semen Brassicae campestris of isozygotying is 60～80mg/kg, and the normal concentration of cross-bred rape seed is 80～120mg/kg.
In choosing the step of kind of primary dcreening operation plant, before the pretreatment (pretreatment of the sterilizing namely carried out in setting screening criteria concentration) that seed carries out above-mentioned sterilizing, pretreatment can also include: by seed dry in the sun 20～30h at 20～35 DEG C.So, it is possible to improve the germination percentage of seed.After the pretreatment carrying out sterilizing, seed is put in the culture dish being lined with 2～3 metafiltration paper, setting up repetition and comparison, the cadmium solution (heavy metal screening solution) of dropping normal concentration in culture dish (culture dish except control experiment), then carrying out germinates cultivates.Certainly, the condition that the condition that this germination is cultivated also cultivates (germination namely carried out in setting screening criteria concentration is cultivated) with above-mentioned germination is identical.Again when germination, calculate germination percentage and seedling-erecting ratio.
Germination percentage=(chitting piece number/seed sum) 100%,
Seedling-erecting ratio=(upright Seedling number/chitting piece number) 100%.
During selection, regardless of whether with reference to germination percentage, can choose seedling-erecting ratio high front 10～20% the kind that plant variety is primary dcreening operation plant corresponding to seed because seedling-erecting ratio is high, then in general, germination percentage is also high.
In the kind of screening super enrichment heavy metal plant, the heavy metal concentration of many parts of soil is the concentration processed through Concentraton gradient, and process range is.
Concentraton gradient can be established as: 10mg/kg, 20mg/kg, 40mg/kg, 60mg/kg, 80mg/kg, 100mg/kg, and for example 75mg/kg, 80mg/kg, 85mg/kg, 90mg/kg, 95mg/kg, 100mg/kg, 105mg/kg.Caddy (Cleary) 2/5 hydrate soln and/or cadmium nitrate solution is utilized to process, it is preferable that to utilize Caddy (Cleary) 2/5 hydrate soln to process.Caddy (Cleary) 2/5 hydrate can be weighed, be dissolved in above-mentioned process water, according to concentration needs, make solution, it is possible to use aerosol apparatus, by the spray solution of variable concentrations in soil, make the many parts of soil that heavy metal concentration is different.It is noted that in order to avoid the accuracy of later experiments so that each position content of beary metal of the plant grown up to is uniform, so needing uniform to heavy metal solution and mixing with soil.And in order to realize this purpose, it is possible to before soil mixes with solution, first make soil through air-dry, sort, pick, levigate, sieve dry, pulverize, sieving etc. processes, it is ensured that the breathability of soil and mellowness so that it is better mixed homogeneously with solution.
Before the planting seed of the kind of each primary dcreening operation plant, it is also possible to carry out pretreatment, carry out dry in the sun process by seed, to improve the germination percentage of seed.Then sow, then the mass ratio that every part of soil applies fertilizer, fertilizer and soil is 5:7000～8000.Fertilizer can provide the nutrient that one or more plants are required, and soil property can be improved, level of increasing soil fertility.In fertilizer, nitrogen content is 12～18%, and phosphorus content is 12～18%, and potassium content is 12～18%.Before fertilising, it is possible to be dissolved in a small amount of water by fertilizer, make fertilizer dissolve, form fertilizer solution, spray fertilizer solution and apply fertilizer, so, apply fertilizer more uniform.Then carry out normal field management, until seed growth is plant, then measure cadmium content.Measuring cadmium content can be further: respectively before plant bolting and after plant blossom sets seeds, and measures the content of beary metal of the root of the content of beary metal of every part of soil and plant corresponding to every part of soil and the content of beary metal of plant shoot.So, by detecting, it is possible to determine that the Brassica campestris L in which period is higher to the bioaccumulation efficiency of cadmium, because the essentialspecies plant of certain areas is not Brassica campestris L, if bioaccumulation efficiency is relatively higher during Brassica campestris L bolting, then according to plant growing situation, illustrate that Brassica campestris L just has been able to reach the purpose of rehabilitating soil in seedling stage, then can before Brassica campestris L bolting, just pull out Brassica campestris L, continue the crop of lower season of plantation this area, so, the normal plantation of these area crops would not be affected.It addition, after plant blossom is set seeds, it is also possible to measure the content of beary metal of the seed that every part of plant corresponding to soil is born simultaneously.By this test, it is determined that cadmium content in the Semen Allii Tuberosi born, thus determining the purposes of Semen Allii Tuberosi, if cadmium content is less than food sanitation standard (cadmium content≤0.2mg/kg) in Semen Allii Tuberosi, then the oil squeezed out may be used for eating；If cadmium content exceedes food sanitation standard in Semen Allii Tuberosi, then the oil squeezed out may be used for bio-power plant.Selection standard finally according to hyperaccumulative plant or super enrichment heavy metal plant, it is determined that the kind of super enrichment heavy metal plant.
Wherein, when measuring cadmium content, it is possible to adopt the methods such as atomic absorption spectrography (AAS) to measure.And after the kind filtering out super enrichment heavy metal plant, it is possible to the growth characteristic according to kind, carry out demonstrating than more serious area in cadmium pollution, promotion and application, verify its concentration effect to cadmium.
Above, Brassica campestris L can select the cabbage type rape of extensively plantation to test.
Below in conjunction with embodiment, inventive feature and performance are described in further detail:
Embodiment one
Embodiments of the invention one provide a kind of super enrichment Brassica campestris L screening technique, comprise the following steps,
A. select 400 seeds, utilize the sodium hypochlorite of 4g/kg to soak seed 8min, then pour out sodium hypochlorite, continuously with pure water by wash seeds 2 times；
B. utilize Caddy (Cleary) 2/5 hydrate and pure water, set up the cadmium test solution of 10mg/kg, 20mg/kg, 40mg/kg, 60mg/kg, 80mg/kg, 100mg/kg, 120mg/kg7 Concentraton gradient；
C. 400 Semen Brassicae campestris are divided into 8 parts, it is individually positioned in 8 culture dishs, number consecutively is 1～No. 8,50 Semen Brassicae campestris of each culture dish, wherein 1～No. 7 culture dish drips the cadmium test solution that 3ml concentration is 10～120mg/kg successively, retain the clear water of No. 8 culture dish dropping equivalent, as a control group, set up 2 repetitions；
D. all culture dishs of step c being placed in 22 DEG C, periodicity of illumination is bright phase 15h, in the environment of dark phase 9h, carries out germinateing and cultivates, and the 6th day statistics seedling-erecting ratio, the concentration of the cadmium test solution choosing No. 4 culture dish droppings is screening criteria concentration；
E. select blue or green oil 14, blue or green oil 331, Zhejiang oil 601, in double; two No. 4, in double; two No. 8, in double; two No. 9, in double; two No. 10, in double; two No. 11, rich oil 6, Zhejiang oil 50, Zhejiang oil 601, Zhejiang oil 50, oil grind No. 10,100 rape varieties such as middle oily 6766, holy light 402, river oil 36, river oil 37, China's oil assorted 12, assorted No. 6 of China's oil, oil 8,50 seeds of each variety adoption, all adopt the method for step a to carry out pretreatment by the Semen Brassicae campestris of each kind；Then the seed of pretreated 100 rape varieties is put in 100 culture dishs, the screening criteria concentration that each culture dish dropping 3ml step d confirms, set up comparison and 2 repetitions, again all culture dishs are placed in 22 DEG C, periodicity of illumination is bright phase 15h, in the environment of dark phase 9h, carry out germinateing and cultivate；The seedling-erecting ratio of the seed adding up each rape variety on the 6th day, select seedling-erecting ratio high front 10% rape variety corresponding to seed be primary dcreening operation rape variety, be oil grind 817, in 10 kinds such as double; two No. 11；
nullF. Caddy (Cleary) 2/5 hydrate and pure water are utilized，Preparation cadmium solution，Then 10 groups of Culture basins are taken，It is designated as the 1st group～the 10th group，Often 7 Culture basins of group，Label 1～7，Often group takes 6 parts of soil，Cadmium solution is sprayed respectively in every a soil，Mix homogeneously，Make the cadmium solution concentration respectively 10mg/kg of 6 parts of soil、20mg/kg、40mg/kg、60mg/kg、80mg/kg、100mg/kg，Then by 6 parts of Homogeneous phase mixing, the soil of cadmium solution loads in 6 Culture basins，According to 10mg/kg、20mg/kg、40mg/kg、60mg/kg、80mg/kg、The concentration order of 100mg/kg loads in 1～No. 6 Culture basin，Take 1 part of soil again，Toward wherein spraying isopyknic aqueous solution，It is then charged in No. 7 Culture basins，As a control group；
G., the seed that oil grinds 817 10 kinds such as grade is all divided into 7 parts, every part of 50 seeds, in conventional seed planting season, it is seeded in the Culture basin of the 1st group～the 10th group respectively, then field management is carried out, until seed is all grown to rapeseed plants, measure the cadmium content of soil in each Culture basin again, the cadmium content of the root of the rapeseed plants that every part of soil is corresponding and the cadmium content of plant shoot, the selection standard of super enrichment Brassica campestris L is contrasted according to the cadmium content before and after soil remediation, learn that double, two No. 11 kinds make the cadmium content in soil reduce, and meet the selection standard of super enrichment Brassica campestris L, it is defined as super enrichment rape variety.
Embodiment two
Embodiments of the invention two provide a kind of super enrichment heavy metal foliage filter screening method, comprise the following steps,
A. select 315 seeds, utilize the potassium permanganate solution of 6g/kg to soak seed 12min, then pour out potassium permanganate solution, continuously with pure water by wash seeds 4 times；
B. utilize cadmium nitrate and ultra-pure water, set up the cadmium test solution of 30mg/kg, 50mg/kg, 70mg/kg, 90mg/kg, 110mg/kg, 130mg/kg6 Concentraton gradient；
C. 315 Semen Brassicae campestris are divided into 7 parts, it is individually positioned in 7 culture dishs, number consecutively is 1～No. 7,45 Semen Brassicae campestris of each culture dish, wherein 1～No. 6 culture dish drips the cadmium test solution that 5ml concentration is 30～130mg/kg successively, retain the clear water of No. 7 culture dish dropping equivalent, as a control group, set up 3 repetitions；
D. all culture dishs of step c being placed in 25 DEG C, periodicity of illumination is bright phase 17h, in the environment of dark phase 7h, carries out germinateing and cultivates, and the 8th day statistics seedling-erecting ratio, the concentration of the cadmium test solution choosing No. 5 culture dish droppings is screening criteria concentration；
E. select wide source 58, contain Hunan oil 9, oil grind No. 10, moral oil 158, kernel oil 202, China's oil assorted No. 12, normal assorted oily No. 2, assorted No. 62 of China's oil, China's Hunan oil 10, holy light 77, Keeping high quality, assorted No. 15 of China's oil, Zhejiang oil 28, oil grind No. 817, China's Hunan oil 12, moral oil 5, spring cloud 763, Deyou 8, assorted No. 108 of rich oil, 150 rape varieties such as Feng oil 737,45 seeds of each variety adoption, all adopt the method for step a to carry out pretreatment by the Semen Brassicae campestris of each kind；Then the seed of pretreated 150 rape varieties is put in 150 culture dishs, the screening criteria concentration that each culture dish dropping 5ml step d confirms, set up comparison and 3 repetitions, again all culture dishs are placed in 25 DEG C, periodicity of illumination is bright phase 17h, in the environment of dark phase 7h, carry out germinateing and cultivate；The seedling-erecting ratio of the seed adding up each rape variety on the 8th day, select seedling-erecting ratio high front 20% rape variety corresponding to seed be primary dcreening operation rape variety, namely oil grind No. 10, Feng oil 737, assorted No. 62 of China's oil, 30 rape varieties such as moral oil 5；
nullF. cadmium nitrate and ultra-pure water are utilized，Preparation cadmium solution，Then 30 groups of Culture basins are taken，It is designated as the 1st group～the 30th group，Often 8 Culture basins of group，Label 1～8，Often group takes 7 parts of soil，Cadmium solution is sprayed respectively in every a soil，Mix homogeneously，Make the cadmium solution concentration respectively 30mg/kg of 7 parts of soil、50mg/kg、70mg/kg、90mg/kg、110mg/kg、130mg/kg、150mg/kg，Then by 7 parts of Homogeneous phase mixing, the soil of cadmium solution loads in 7 Culture basins，According to 30mg/kg、50mg/kg、70mg/kg、90mg/kg、110mg/kg、130mg/kg、The concentration order of 150mg/kg loads in 1～No. 7 Culture basin，Take 1 part of soil again，Toward wherein spraying isopyknic aqueous solution，It is then charged in No. 8 Culture basins，As a control group；
G., the seed that oil grinds 30 kinds such as No. 10 grades is all divided into 8 parts, every part of 45 seeds, in conventional seed planting season, it is seeded in the Culture basin of the 1st group～the 30th group respectively, then each Culture basin is applied fertilizer, the mass ratio of the soil in fertilizer and Culture basin is 5:8000, carry out field management again, until seed is all grown to rapeseed plants, measure the cadmium content of soil in each Culture basin again, the cadmium content of the root of the rapeseed plants that every part of soil is corresponding and the cadmium content of plant shoot, the selection standard of super enrichment Brassica campestris L is contrasted according to the cadmium content before and after soil remediation, learn Feng oil 737, the kind of moral oil 5 makes the cadmium content in soil reduce, and meet the selection standard of super enrichment Brassica campestris L, it is defined as super enrichment rape variety.
Embodiment three
Embodiments of the invention three provide a kind of super enrichment heavy metal foliage filter screening method, comprise the following steps,
A. in selecting, 400 seeds of double; two No. 11, the alcohol-pickled seed 5min utilizing mass concentration to be 75%, pour out ethanol, sodium hypochlorite followed by 5g/kg soaks seed 10min, pour out sodium hypochlorite, then with pure water by wash seeds 3 times, then clean 1 time with ultra-pure water；
B. utilize Caddy (Cleary) 2/5 hydrate and pure water, set up the cadmium test solution of 10mg/kg, 20mg/kg, 40mg/kg, 60mg/kg, 80mg/kg, 100mg/kg, 120mg/kg7 Concentraton gradient；
C. 400 Semen Brassicae campestris are divided into 8 parts, it is individually positioned in 8 culture dishs being lined with 2 metafiltration paper, number consecutively is 1～No. 8,50 Semen Brassicae campestris of each culture dish, wherein 1～No. 7 culture dish drips the cadmium test solution that 4ml concentration is 10～120mg/kg successively, retain the clear water of No. 8 culture dish dropping equivalent, as a control group, set up 2 repetitions；
D. all culture dishs of step c being placed in 23 DEG C, periodicity of illumination is bright phase 16h, in the environment of dark phase 8h, carries out germinateing and cultivates, within the 7th day, add up germination percentage and seedling-erecting ratio, and the concentration of the cadmium test solution choosing No. 5 culture dishs droppings is screening criteria concentration, i.e. 80mg/kg；
E. select assorted No. 10 of moral oil, assorted No. 1 of south oil, China's Anhui oil 2, happy No. 1 of section, Deyou 8, assorted No. 14 of No. 5 China's oil of moral oil, assorted No. 16 of China's oil, assorted No. 695 of Hunan, Hunan oil 17, in double; two No. 4, in double; two No. 8, in 200 rape varieties such as double; two No. 11, including 156 conventional varieties (kind of isozygotying) and 44 Hybrids, 50 seeds of each variety adoption；
F. all seeds of all kinds of step e are placed at 28 DEG C and shine 24h, then all seeds all adopt the alcohol-pickled method cleaned to ultra-pure water in step a carry out pretreatment；Again the seed of pretreated 200 rape varieties is put in 200 culture dishs being lined with 2 metafiltration paper, the screening criteria concentration that each culture dish dropping 4ml step d confirms, set up comparison and 2 repetitions, again all culture dishs are placed in 23 DEG C, periodicity of illumination is bright phase 16h, in the environment of dark phase 8h, carry out germinateing and cultivate；The germination percentage of the seed adding up each rape variety on the 7th day and seedling-erecting ratio, using germination percentage as select while reference seedling-erecting ratio high before 10% rape variety corresponding to seed for primary dcreening operation rape variety, be Hunan oil 15, double; two No. 3 of China, in double; two No. 11,20 kinds such as moral oil 5；
F. utilize Caddy (Cleary) 2/5 hydrate and pure water, prepare cadmium solution, then take 20 groups of Culture basins, be designated as the 1st group～the 20th group, often 7 Culture basins of group, label 1～7；
nullOften group takes 6 parts of soil，Every part of soil first passes through air-dry、Sorting、Pick、Levigate、Sieve and dry、Pulverize、Sieve process，Then in every a soil, cadmium solution is sprayed respectively，Mix homogeneously，Make the cadmium solution concentration respectively 10mg/kg of 6 parts of soil、20mg/kg、40mg/kg、60mg/kg、80mg/kg、100mg/kg，Then by 6 parts of Homogeneous phase mixing, the soil of cadmium solution loads in 6 Culture basins，According to 10mg/kg、20mg/kg、40mg/kg、60mg/kg、80mg/kg、The concentration order of 100mg/kg loads in 1～No. 6 Culture basin，Take 1 part of soil again，Toward wherein spraying isopyknic aqueous solution，It is then charged in No. 7 Culture basins，As a control group，2 repetitions are set；
G. by the seed of double; two 20 kinds such as No. 11 grades be all placed at 28 DEG C solarization 24h, then in conventional seed planting season, it is seeded in the Culture basin of the 1st group～the 20th group respectively, then each Culture basin is applied fertilizer, the mass ratio of the soil in fertilizer and Culture basin is 5:7000, nitrogenous 15% in fertilizer, phosphorous 15%, containing potassium 15%；
Carry out field management again, until seed is all grown to rapeseed plants, the cadmium content of soil in each Culture basin is measured before bolting, the cadmium content of the root of the rapeseed plants that every part of soil is corresponding and plant shoot (stem, leaf) cadmium content, the cadmium content of soil in each Culture basin is again measured after blossoming and having seeds, the cadmium content of the root of the rapeseed plants that every part of soil is corresponding and plant shoot (stem, leaf), the cadmium content of the seed that Brassica campestris L is born, the selection standard of super enrichment Brassica campestris L is contrasted according to the cadmium content before and after soil remediation, learn that moral No. 5 kinds of oil make the cadmium content in soil reduce, and meet the selection standard of super enrichment Brassica campestris L, it is defined as super enrichment rape variety.
In the present embodiment, cadmium detection accepted standard: root and straw accepted standard are GB/T13082-91AAS, and soil accepted standard is USEPAMETHOD, and seed accepted standard is GB5009.15-2014GFAAS, it is determined that Brassica campestris L super enrichment ability.
In prior art, general 200m2Booth carry out potted plant, cost is high, it is necessary to certain floor space, and management and screening cost are high, it is necessary to planting for 3 generations just can complete screening, and the cycle needs 2～3 years.And utilizing the method that the present embodiment provides to screen, the cycle, below 2 years, reduces by more than 70% in management and on cost.
The foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, for a person skilled in the art, the present invention can have various modifications and variations.All within the spirit and principles in the present invention, any amendment of making, equivalent replacement, improvement etc., should be included within protection scope of the present invention.
Claims (10)
1. the screening technique of the kind of a super enrichment heavy metal plant, it is characterised in that comprise the following steps:
Set screening criteria concentration: be respectively put into by many seeds of the plant through pretreatment in multiple Heavy metal solution that concentration is different, carry out germinateing and cultivate, the concentration choosing the Heavy metal solution at the seed place of the described plant that seedling-erecting ratio is 50% is described screening criteria concentration, described seedling-erecting ratio=(B/C) 100%, B is upright Seedling number, and C is germination number；
Choose the kind of primary dcreening operation plant: select multiple kinds of described plant, seed by each described kind through pretreatment, put in the heavy metal screening solution with described screening criteria concentration, carry out germinateing and cultivate, calculate the seedling-erecting ratio of the seed of each described kind, select seedling-erecting ratio high front 10～20% the kind that described kind is described primary dcreening operation plant corresponding to described seed；And
The kind of screening super enrichment heavy metal plant: the seed of the kind of each described primary dcreening operation plant through pretreatment is seeded in the many parts of soil that heavy metal concentration is different respectively, and carry out field management, until it is grown to plant, measure the content of beary metal of the root of the content of beary metal of every part of described soil and described plant corresponding to every part of soil and the content of beary metal of plant shoot, when described plant is survived, and the content of beary metal of its plant shoot is higher than the content of beary metal of root, and the substance withdrawl syndrome of its plant shoot is more than the substance withdrawl syndrome of root, and the content of beary metal before the soil remediation at its place more than administer after content of beary metal time, determine the kind that the kind belonging to described plant is super enrichment heavy metal plant.
2. the screening technique of the kind of super enrichment heavy metal plant according to claim 1, it is characterized in that, in the step of the described kind choosing primary dcreening operation plant, the seed of each described plant variety of pretreatment includes: by seed dry in the sun 20～30h at 20～35 DEG C；In the step of described screening enriching heavy metal plant variety, the described seed of each described primary dcreening operation plant variety of pretreatment includes: by described seed dry in the sun 20～30h at 20～35 DEG C.
3. the screening technique of the kind of super enrichment heavy metal plant according to claim 1, it is characterised in that described plant is Brassica campestris L, and described heavy metal is cadmium.
4. the screening technique of the kind of super enrichment heavy metal plant according to claim 3, it is characterised in that described heavy metal solution is at least one in Caddy (Cleary) 2/5 hydrate soln, cadmium nitrate solution, it is preferable that Caddy (Cleary) 2/5 hydrate soln.
5. the screening technique of the kind of super enrichment heavy metal plant according to claim 1, it is characterised in that in the step of the kind of described screening super enrichment heavy metal plant, the heavy metal concentration of described many parts of soil is 0.3～150mg/kg.
6. the screening technique of the kind of super enrichment heavy metal plant according to claim 1, it is characterized in that, in the step of the kind of described screening super enrichment heavy metal plant, measuring content of beary metal is further: respectively before described plant bolting and after blossoming and having seeds, and measures the content of beary metal of the root of the content of beary metal of every part of described soil and described plant corresponding to every part of described soil and the content of beary metal of plant shoot respectively.
7. the screening technique of the kind of super enrichment heavy metal plant according to claim 6, it is characterised in that after described plant is blossomed and had seeds, also needs to measure the content of beary metal of the seed that every part of described plant corresponding to described soil is born.
8. the screening technique of the kind of super enrichment heavy metal plant according to claim 1, it is characterized in that, in the step of the kind of described screening super enrichment heavy metal plant, after described sowing, before carrying out described field management, also include: the mass ratio that soil every part described applies fertilizer, described fertilizer and described soil is 5:7000～8000.
9. the screening technique of the kind of super enrichment heavy metal plant according to claim 8, it is characterised in that in described fertilizer, nitrogen content is 12～18%, and phosphorus content is 12～18%, and potassium content is 12～18%.
10. the screening technique of the kind of super enrichment heavy metal plant according to claim 1, it is characterized in that, in the step of described setting screening criteria concentration and the step of the described kind choosing primary dcreening operation plant, the described condition cultivated of germinateing is: temperature 22～25 DEG C, periodicity of illumination is bright phase 15～17h, dark phase 7～9h.
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