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Method for repairing heavy metal and organic compound contaminated soil using transgenic plant

A technology of transgenic plants and compound pollution, which is applied in the field of heavy metal and organic compound pollution soil remediation, and can solve the problem of arduous task of remediation of compound pollution.

Inactive Publication Date: 2008-11-26
QINGDAO UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0015] Combined pollution of heavy metals and organics exists in many areas, and combined pollution has more serious impacts, so the task of remediating combined pollution is more difficult

Method used

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  • Method for repairing heavy metal and organic compound contaminated soil using transgenic plant
  • Method for repairing heavy metal and organic compound contaminated soil using transgenic plant
  • Method for repairing heavy metal and organic compound contaminated soil using transgenic plant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Construction and transformation of the plant binary expression vector of embodiment 1 Agrobacterium

[0037] The human-derived cytochrome P450 2 E1 gene and the γ-glutamylcysteine ​​synthetase (γ-ECS) gene derived from Escherichia coli were digested with restriction endonucleases BamH1 and Sac1, and connected to the plant The binary transformation vector pCAMBIA2301G, and the obtained recombinant expression vector was named pCAMBIA2301G-p450-ECS.

[0038] The recombinant expression vector pCAMBIA2301G-p450-ECS was transformed into Agrobacterium tumefaciens strain LBA4404 by electroporation.

Embodiment 2

[0039] Embodiment 2 plant transformation

[0040] The recombinant expression vector pCAMBIA2301G-p450-ECS was transformed into the leaves of alfalfa by Agrobacterium-mediated leaf disk transformation method.

[0041] Centrifuge the Agrobacterium culture medium with pCAMBIA2301G-p450-ECS plasmid cultivated overnight, suspend the bacteria in 10ml of 0.15mol / L NaCL, collect the bacteria by centrifugation, and resuspend in 1ml of 20mmol / L CaCL 2 Suspend the solution, cut alfalfa leaves into small pieces of 0.5X0.5cm, soak them in the bacterial solution containing the pCAMBIA2301G-p450-ECS plasmid for 20min, blot the excess bacterial solution with filter paper, and place it on MS medium for co-cultivation 3 days, then transfer to the screening medium (MS+0.5mg / L6-BA+0.5mg / L 2,4-D+20mg / L Kanamycin+300mg / L Cefixime) containing kanamycin for preliminary screening . When the bud grows to about 1 cm, the leaf disk of the long bud is transferred to the rooting medium (MS+70mg / LKanamyci...

Embodiment 3

[0042] Example 3 Remediation experiment of transgenic alfalfa to cadmium, lead-TCE compound polluted soil

[0043] First, the heavy metals cadmium and lead were dissolved in their inorganic salt solution (CdCl 2 5H2O, PbCl 2 , analytically pure) was uniformly applied to the soil, and two high and low treatment concentrations were set for cadmium, lead and TCE respectively (the cadmium was: 0.5, 1.5 mg·kg -1 , Lead: 100, 300mg·kg -1 , TCE is 10, 100mg·kg -1 ), using L2 3 Orthogonal design, and the following controls were set up: (1) non-pollutant soil planted with transgenic alfalfa (2) polluted soil planted with wild-type alfalfa, with 3 replicates for each treatment.

[0044] The test took the following steps to mix TCE with the test soil:

[0045] ① For each concentration treatment, first dissolve the TCE required for the test with an appropriate amount of acetone, and then add it to the soil that is 1 / 10 of the total amount of soil required for the test design, and sti...

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Abstract

The invention discloses a method for restoring soil compositely polluted by heavy metals and organics by using a transgenic plant and is technically characterized in that foreign genes which can strengthen the efficiency for restoring the organic pollution of the plant and foreign genes which can improve the tolerance and the accumulation to a plurality of metals of the plant are introduced to the plant so as to establish a transgenic plant which has a better restoring effect on the pollution by the heavy metals and the organics. The method of the invention has a better restoring effect on the polluted soil of the three types as follows: (1) the soil polluted by organic pollutants of halohydrocarbon compounds such as trichloroethylene, dibromethane, and the like; (2) the soil polluted by various heavy metals such as Cd, Cr, Cu, Mn, Zn and Pb; (3) the soil compositely polluted by the heavy metals and the organics. In addition, the method of the invention comprises the procedures of establishing the transgenic plant, the screening and cultivation of light-demanding plants and the restoring of polluted soil by the transgenic plant. The method of the invention has broad application prospect in the restoring of the soil compositely polluted by the heavy metals and the organics.

Description

1. Technical field [0001] The invention relates to the fields of biotechnology and environmental protection, in particular to a method for introducing exogenous genes into complex pollution-resistant plants and applying it to remediation of heavy metal and organic compound polluted soil. 2. Background technology [0002] With the development of urbanization, industrialization, automobile exhaust emission and the intensive development of agriculture, people pay more and more attention to environmental issues, and soil pollution caused by heavy metals and organic matter has increasingly become a hot issue for environmental and soil scientists. After the soil is polluted by heavy metals and organic matter, it not only directly affects the yield and quality of crops, but also affects human health and safety through the food chain. The strategy of using green plants to remove pollutants, that is, phytoremediation technology, has the characteristics of low cost, environmental frie...

Claims

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Application Information

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IPC IPC(8): B09C1/00B09C1/10C12N15/09C12N15/82C12N15/12C12N15/29
Inventor 张媛媛刘均洪邵宏波郑成超李凤梅李俊峰苏忠亮初立业王繁业吴汝林
Owner QINGDAO UNIV OF SCI & TECH
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