CN102161976A - Endophyte for promoting growth of plants and remediation of heavy metal-contaminated soil and application - Google Patents
Endophyte for promoting growth of plants and remediation of heavy metal-contaminated soil and application Download PDFInfo
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- CN102161976A CN102161976A CN 201010609930 CN201010609930A CN102161976A CN 102161976 A CN102161976 A CN 102161976A CN 201010609930 CN201010609930 CN 201010609930 CN 201010609930 A CN201010609930 A CN 201010609930A CN 102161976 A CN102161976 A CN 102161976A
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Abstract
The invention provides an endophyte for promoting the growth of plants and the remediation of heavy metal-contaminated soil and application thereof. Bacterial liquid of endophyte SLS18 is utilized to dip-dye Sorghum bicolor L. in a mode of seed bacteria dipping to colonize the endophyte SLS18 into the Sorghum bicolor L. to accompany each stage of the growth of the Sorghum bicolor L., so that the growth of the Sorghum bicolor L. on heavy metal-contaminated marginal land and the remediation of soil are promoted. The endophyte SLS18 is separated from the stem of a hyperaccumulator, namely pokeberry root, is obtained through screening, and is an endophyte which can be successfully colonized in the Sorghum bicolor L., has a remarkable effect of promoting the growth of the Sorghum bicolor L. and has the heavy metal resistance. A method is easy to operate, and is safe and economic, no new pollution is introduced into soil environment, heavy metal-contaminated marginal land resources can be effectively utilized so as to avoid the rival of energy crops and food crops against finite cultivated land, and heavy metal-contaminated land can also be effectively remediated.
Description
Technical field
The invention belongs to the heavy metal soil pollution and repair the field, be specifically related to a kind of endophyte that can promote the energy crop growth, and infect plant, and improve the application method of phytoremediation heavy metal soil pollution efficient.
Background technology
Along with the development of modernization industry, heavy metal contamination has caused serious threat to the human health and the ecosystem, becomes one of global environmental problem gradually.Wherein, manganese can accumulate and cause manganese poisoning, for example parkinsonism, reproductive system and immunity system pathology in human body.And another kind of heavy metal cadmium has carcinogenic, teratogenesis, mutagenesis, and accumulation can cause bone, digestive tube, vascular lesion in human body.Characteristics such as everyone knows, the appliable plant recovery technique is removed the heavy metal in the soil, compares with traditional physical chemistry recovery technique to have lot of advantages, and for example cost is low, safe and effective, environmentally friendly.Especially use super accumulation plant and carry out phytoremediation, this kind of plant not only can not be subjected to the murder by poisoning of heavy metal in process of growth, can also accumulate plurality of heavy metal in its body.Yet super accumulation plant because biomass bioavailability low and heavy metal in soil is low, has been limited the phytoremediation efficient of super accumulation plant when the growth of heavy metal contamination area.Therefore, many investigators are devoted to study the method that improves super accumulation phytomass.Wherein, the long endophyte of plant combined plant growth-promoting (PGPE) technology has effectively promoted the phytoremediation application and development of super accumulation plant.As everyone knows, PGPE can assist its host plant opposing heavy metal to accumulate in vivo to cause toxic effect, can promote the growth of host plant simultaneously.This is because PGPE has several plant growth-promoting lead aircraft systems: (1) promotes plant-growth by producing indolylacetic acid plant hormones such as (IAA); (2) assist plant from soil, to obtain the required iron ion of growth by producing siderophore, thereby make host plant obtain optimum growth conditions; (3), thereby reduce the restraining effect of pressure ethene to plant-growth by the active concentration that reduces pressure ethene precursor ACC of acc deaminase.
Yet from another angle, what be worth arousing attention is to utilize these promotes growth mechanism of PGPE to promote the growth of energy crop on the marginal soil of heavy metal contamination, thereby improves the biomass of energy crop and the CO that catches from air
2Raw material as bioenergy is very significant behave.Described marginal soil is meant because of arid, saline and alkaline is high, be subjected to heavy metal and Organic pollutants etc. to be not suitable for the soil of general crop growth.Therefore this behave can be avoided the competition to agricultural resources such as limited arable lands of energy crop and food crop.Along with the increase day by day that bioenergy uses, the problem that energy crop takies the food crop arable land has become one of socioeconomic problem that needs to be resolved hurrily.According to the report of up-to-date economic analysis,, be acknowledged as at present in the world one of energy crop of potentialization because sweet sorghum (Sorghum bicolor L.) has the photosynthetic efficiency height, produces and drop into low, characteristics such as biomass is high, drought-resistant, salt tolerant.In addition, sweet sorghum also can extract the heavy metal composition in the soil, has the potentiality that are used for phytoremediation.Yet as far as we know, also do not have patent and bibliographical information to cross the separating obtained promotes growth of anti-heavy metal endophyte in the super accumulation plant materials at present, be applied to promote growth and the phytoremediation of energy crop on the marginal land of heavy metal contamination.
Summary of the invention
The invention provides a kind of endophyte that can promote the energy crop growth, and infect plant, and improve the application method of phytoremediation heavy metal soil pollution efficient.The present invention is simple to operate, safety, economy, and edatope is not introduced new pollution, can effectively avoid the competition to limited arable land of energy crop and food crop, simultaneously the soil of repairing heavy metal pollution effectively.
For addressing the above problem, technical scheme provided by the invention is:
A kind of endophyte that promotes that plant-growth and heavy-metal contaminated soil are repaired, it is characterized in that, described endophyte is bacillus sp.SLS18, on December 20th, 2010, Chinese typical culture collection center in Chinese Wuhan City, Hubei Province Wuhan University, carried out preservation, deposit number is CCTCCNO:M 2010359.
Described endophyte is applied to promote plant-growth and heavy-metal contaminated soil reparation; Specifically be that described endophyte SLS18 is contaminated sweet sorghum seed, the sweet sorghum sowing is grown on heavy metal (as: Mn/Cd) contaminated land then, carries out soil remediation.The bacterial concentration of endophyte SLS18 is 10
9Cfu mL
-1Soak 4h in the bacterium liquid.
Described sweet sorghum seed carries out surface sterilization before soaking bacterium process is: sweet sorghum seed is soaked in 70% ethanolic soln 1min, be soaked in 1% chlorine bleach liquor 10min again, with after the rinsed with deionized water after the sterilization at least 5 times, dry and treat that seed soaks bacterium subsequently.
Sweet sorghum seed soaks the used endophyte SLS18 thalline of bacterium process for being in logarithmic phase, the endophyte culturing process is: with the SLS18 bacterial classification inoculation of 100 μ L preservations to the sterile LB substratum of 25mL, after nutrient solution placed 30 ℃ of shaking table concussions to cultivate 24h, preparation concentration was 10
9Cfu mL
-1Bacterium liquid.
Preparation concentration is 10
9Cfu mL
-1The preparation process of bacterium liquid be: get 5mL and cultivate endophyte bacterium liquid behind the 24h at 13000r/min, centrifugal under the condition of 10min, with centrifugal again after the washed with de-ionized water, then the thalline of collecting is suspended in 0.03M MgSO
4In the solution, and be diluted to the OD that surveys
600Be 1, promptly getting corresponding concentration is 10
9Cfu mL
-1Bacterium liquid.
The sweet sorghum seed of contaminating endophyte SLS18 is seeded in the heavy metal Mn/Cd Contaminated soil, the SLS18 endophyte is survived in the sweet sorghum body, follow each stage of sweet sorghum growth, promote the growth of sweet sorghum on the heavy metal contamination marginal land and the purpose of soil remediation thereby reach.
The anti-heavy metal bacterium that above-mentioned promotes growth endophyte SLS18 obtains for separation in the super accumulation plant pokeweed body, and can successfully contaminate in the sweet sorghum body.
The screening and separating process of bacterial strain of the present invention and method are with reference to a patent " a kind of method that obtains endophyte of plant of the separating " (application number: 200810107514.1) of this scientific research group.Its colony characteristics on the LB solid medium is: size 3 is to 6mm; Circular, neat in edge; Milk yellow; Smooth surface, protuberance, opaque.Its part Physiology and biochemistry character is accredited as: Gram-negative bacteria; Produce indolylacetic acid and siderophore albumen; Its heavy metal resistance specifically shows as with MIC (minimum inhibition concentration): Mn 2000mg/L, Cd 40mg/L, Pb 500mg/L, Cu 200mg/L, Zn 100mg/L (LB substratum).By 16S rDNA is carried out sequencing analysis, endophyte SLS18 is accredited as bacillus (bacillus sp.)
Advantage of the present invention shows as bright simple to operate safe, economical, edatope is not being introduced under the prerequisite of new pollution, can effectively promote energy crop being subjected to grow the soil of while repairing heavy metal pollution on the marginal land of heavy metal contamination.
Description of drawings
Fig. 1 is an operational flowchart of the present invention;
Fig. 2 is the influence of endophyte SLS18 bacterium to the root dry weight (g) of sweet sorghum, Phytolacca acinosa, black nightshade, and heavy metal-polluted soil concentration is respectively 0mg kg
-1, Mn 2000mg kg
-1, Cd 50mg kg
-1
Fig. 3 is the influence of endophyte SLS18 bacterium to the over-ground part dry weight (g) of sweet sorghum, Phytolacca acinosa, black nightshade, and heavy metal-polluted soil concentration is respectively 0mg kg
-1, Mn 2000mg kg
-1, Cd 50mg kg
-1
Fig. 4 is that endophyte SLS18 is to sweet sorghum, Phytolacca acinosa, (the μ g strain of black nightshade accumulation Mn ability
-1) influence, heavy metal-polluted soil concentration is respectively Mn 2000mg kg
-1
Fig. 5 is that endophyte SLS18 is to sweet sorghum, Phytolacca acinosa, (the μ g strain of black nightshade accumulation Cd ability
-1) influence, heavy metal-polluted soil concentration is respectively Cd 50mg kg
-1
Embodiment:
Be intended to further specify the present invention below in conjunction with embodiment, and unrestricted the present invention.
Endophyte SLS18 is contaminated sweet sorghum simultaneously, and behind Phytolacca acinosa (P.acinosa R.) and black nightshade (S.nigrum L.) seed, sow growth respectively with the three kind of plant seeds of not contaminating endophyte, carry out the test effect contrast, wherein Phytolacca acinosa and black nightshade are the super accumulation of newfound two heavy metal species plant; The test operation process is undertaken by Fig. 1.
3 parallel tests are carried out in all tests, and endophyte SLS18 is significantly improved to the sweet sorghum biomass, are that the statistics of reference is seen Fig. 2, Fig. 3, Fig. 4 with Phytolacca acinosa and black nightshade.
Endophyte SLS18 sees Table 1 to the promotes growth rate of sweet sorghum, Phytolacca acinosa, black nightshade, and heavy metal-polluted soil concentration is respectively 0mg kg
-1, Mn 2000mg kg
-1, Cd 50mg kg
-1
Table 1
Endophyte SLS18 sees Table 2 to the enhancing rate of individual plant sweet sorghum, Phytolacca acinosa, black nightshade over-ground part heavy metal accumulation total amount, and heavy metal-polluted soil concentration is respectively Mn 2000mg kg
-1, Cd 50mg kg
-1
Table 2
Endophyte SLS18 is at sweet sorghum, Phytolacca acinosa, intravital situation (the cfu g that grows surely of black nightshade
-1The plant fresh weight) see Table 3, heavy metal-polluted soil concentration is respectively 0mg kg
-1, Mn 2000mg kg
-1, Cd 50mg kg
-1
Table 3
Improved to significant effective of the present invention biomass and the individual plants heavy metal integral dose of sweet sorghum, Phytolacca acinosa, black nightshade, endophyte SLS18 can successfully grow in the sweet sorghum body surely, and the acting in conjunction of SLS18 associating sweet sorghum is stronger than Phytolacca acinosa and black nightshade.Therefore, the present invention can realize promoting the application of energy crop in heavy metal contamination geographic growth in limit and heavy metal reparation, thereby effectively utilize the marginal soil of heavy metal contamination, thereby avoid the competition to limited arable land of energy crop and food crop, the while is the soil of repairing heavy metal pollution effectively.
Claims (8)
1. an endophyte that promotes that plant-growth and heavy-metal contaminated soil are repaired is characterized in that described endophyte is bacillus sp.SLS18, and deposit number is CCTCC NO:2010359.
2. the described endophyte of claim 1 is applied to promote plant-growth and heavy-metal contaminated soil reparation.
3. the application method of endophyte according to claim 2 is characterized in that, described endophyte SLS18 is contaminated sweet sorghum seed, and the sweet sorghum sowing is grown on the ground at heavy metal polluted soil then, carries out soil remediation.
4. according to the application method of claim 2 or 3 described endophytes, it is characterized in that, is 10 with concentration
9Cfu mL
-1The bacterium liquid of endophyte SLS18 sweet sorghum seed carried out seed soak bacterium.
5. the application method of endophyte according to claim 4 is characterized in that, described sweet sorghum seed places bacterium liquid to soak 4h.
6. the application method of endophyte according to claim 4, it is characterized in that, described sweet sorghum seed carries out surface sterilization before soaking bacterium process is: sweet sorghum seed is soaked in 70% ethanolic soln 1min, be soaked in 1% chlorine bleach liquor 10min again, with after the rinsed with deionized water after the sterilization at least 5 times, dry and treat that seed soaks bacterium subsequently.
7. the application method of endophyte according to claim 4, it is characterized in that, sweet sorghum seed soaks the used endophyte SLS18 thalline of bacterium process for being in logarithmic phase, the endophyte culturing process is: with the SLS18 bacterial classification inoculation of 100 μ L preservations to the sterile LB substratum of 25mL, after nutrient solution placed 30 ℃ of shaking table concussions to cultivate 24h, preparation concentration was 10
9Cfu mL
-1Bacterium liquid.
8. the application method of endophyte according to claim 7 is characterized in that, preparation concentration is 109cfu mL
-1The preparation process of bacterium liquid be: get 5mL and cultivate endophyte bacterium liquid behind the 24h at 13000r/min, centrifugal under the condition of 10min, with centrifugal again after the washed with de-ionized water, then the thalline of collecting be suspended in the 0.03M MgSO4 solution, and be diluted to the OD that surveys
600Be 1, promptly getting corresponding concentration is 109cfu mL
-1Bacterium liquid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010609930 CN102161976B (en) | 2010-12-28 | 2010-12-28 | Endophyte for promoting growth of plants and remediation of heavy metal-contaminated soil and application |
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|---|---|---|---|
| CN 201010609930 CN102161976B (en) | 2010-12-28 | 2010-12-28 | Endophyte for promoting growth of plants and remediation of heavy metal-contaminated soil and application |
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| CN102161976A true CN102161976A (en) | 2011-08-24 |
| CN102161976B CN102161976B (en) | 2013-02-06 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102513340A (en) * | 2011-12-13 | 2012-06-27 | 沈阳大学 | Method for repairing soils polluted by heavy metal Cd |
| CN106434497A (en) * | 2016-12-12 | 2017-02-22 | 浙江农林大学 | Siderophore high-yielding strain preparation and application of same in remediation of heavy metal contaminated soil |
| CN108620432A (en) * | 2018-04-26 | 2018-10-09 | 河南寰宇环保科技股份有限公司 | The improvement of heavy metal pollution of soil and ecological restoring method |
| CN109554316A (en) * | 2018-12-28 | 2019-04-02 | 南京农业大学 | A kind of biological prosthetic reagent and restorative procedure for promoting plant growth and development and strengthening accumulation Heavy Metals in Soil Contaminated |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101402918A (en) * | 2008-11-20 | 2009-04-08 | 王梦怡 | Separation process for plant endophyte |
| CN101434906A (en) * | 2008-12-19 | 2009-05-20 | 湖南大学 | Separation process for plant endophyte |
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2010
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101402918A (en) * | 2008-11-20 | 2009-04-08 | 王梦怡 | Separation process for plant endophyte |
| CN101434906A (en) * | 2008-12-19 | 2009-05-20 | 湖南大学 | Separation process for plant endophyte |
Non-Patent Citations (2)
| Title |
|---|
| 《农业环境科学学报》 20071231 姜敏等 重金属抗性内生真菌与其宿主植物重金属抗性关系初探 2038-2042 1-8 第26卷, 第6期 * |
| 《微生物学通报》 20090320 曹喆等 一株龙葵内生细菌SDE06去除Cd+2的试验 328-333 1-8 第36卷, 第3期 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102513340A (en) * | 2011-12-13 | 2012-06-27 | 沈阳大学 | Method for repairing soils polluted by heavy metal Cd |
| CN102513340B (en) * | 2011-12-13 | 2014-02-19 | 沈阳大学 | Method for repairing soils polluted by heavy metal Cd |
| CN106434497A (en) * | 2016-12-12 | 2017-02-22 | 浙江农林大学 | Siderophore high-yielding strain preparation and application of same in remediation of heavy metal contaminated soil |
| CN106434497B (en) * | 2016-12-12 | 2019-06-04 | 浙江农林大学 | Siderophore high yield bacteria preparation and its application in terms of Heavy Metals in Soil Contaminated reparation |
| CN108620432A (en) * | 2018-04-26 | 2018-10-09 | 河南寰宇环保科技股份有限公司 | The improvement of heavy metal pollution of soil and ecological restoring method |
| CN109554316A (en) * | 2018-12-28 | 2019-04-02 | 南京农业大学 | A kind of biological prosthetic reagent and restorative procedure for promoting plant growth and development and strengthening accumulation Heavy Metals in Soil Contaminated |
| CN109554316B (en) * | 2018-12-28 | 2020-04-28 | 南京农业大学 | Bioremediation reagent for promoting plant growth and development and strengthening accumulation of heavy metals in polluted soil and remediation method |
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| CN102161976B (en) | 2013-02-06 |
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