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Plant repair method of applying transgenetic plant composite system to polluted soil

A technology of transgenic plants and polluted soil, applied in the field of heavy metal remediation, to achieve the effect of improving absorption and accumulation capacity, strong adaptability and strong resistance

Active Publication Date: 2019-09-27
INST OF GEOGRAPHICAL SCI & NATURAL RESOURCE RES CAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem to be solved by the present invention is: based on the above background, we cloned and synthesized related genes, constructed Znt1 genes expressed by different promoters, transferred them into different plants and expressed them efficiently, and obtained a kind of Znt1 gene that can efficiently repair heavy metals Genetically modified plants for contaminated soils, providing a new avenue for soil remediation

Method used

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  • Plant repair method of applying transgenetic plant composite system to polluted soil
  • Plant repair method of applying transgenetic plant composite system to polluted soil
  • Plant repair method of applying transgenetic plant composite system to polluted soil

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Construction of transformation vectors and transformation of plants:

[0067] 1. Construction of transformation vector:

[0068] The Znt1 gene (derived from the Brassica cerevisiae ( Thlaspi caerulescens ) Znt1 gene, which was retrieved by the applicant on GenBank and obtained Celestia chinensis ( Thlaspi caerulescens ) of the Znt1 cds sequence, which was obtained by chemically synthesizing the Znt1 target gene fragment by Nanjing Yidao Biotechnology Co., Ltd., and then amplified by PCR), digested with restriction endonucleases SmaⅠ and XbaⅠ, and then ligated to The latter plasmid expression vector is the plant transformation vector p3301-121, and named as p3301-121-Znt1 plasmid (see its structure figure 1 ), transformed into Agrobacterium strain EHA105 by freeze-thaw method;

[0069] 2. Plant genetic transformation:

[0070] The Agrobacterium with p3301-121-Znt1 plasmid cultivated overnight was centrifuged, and the bacteria were suspended in liquid MS 1 / 10 medium...

Embodiment 2

[0072] Resistance of transgenic plants to heavy metals lead and cadmium:

[0073] Material selection and experimental procedures: select the leaves of the transgenic Solanum nigrum line with positive PCR as the material, and multiply the selected leaves by the method of plant tissue culture, and antibiotics (50mg / L Kan+250mg / L Kan+250mg / L Rif ) to screen, when the regenerated plant has 5-6 leaves, select the plant with the same size, remove the root and transfer it to the rooting medium containing different cadmium and lead ion concentrations for growth observation. CdSO 4 (10mg / ml ), PbCl 2 (10mg / ml) solution, so that the cadmium ion concentration in the medium is divided into five levels (0, 50, 100,150, 200 µmol / L), and the lead ion concentration is divided into five levels (0, 25, 50, 75, 100 μmol / L), each level was repeated three times, 28°C, 16h light culture, one month later, the stems, leaves and roots were harvested and washed with distilled water, and then the ha...

Embodiment 3

[0075] Accumulation effect of transformed Solanum nigrum on heavy metals under different substrate conditions:

[0076] Soil culture: Transplant p3301-121-Znt1 gene Solanum nigrum and wild-type Solanum nigrum into flowerpots equipped with 500g soil to cultivate and grow for about one month, during which time MS nutrient solution is used to pour once a week, and after one month, start to CdSO with different concentrations of heavy metals added to the soil 4 (10mg / ml) and PbCl 2 (10mg / ml) solution, where Cd 2+ The concentration is divided into five levels (0, 80, 160, 240, 320µmol / L), Pb 2+ Concentrations were divided into five levels (( 0, 50, 100, 150, 200µmol / L ), and each level was repeated three times. Solanum nigrum was cultivated and grown for one month, and the aboveground stems, leaves and roots were harvested separately, washed with distilled water, and then Dry it at 80°C and pulverize it for testing. The analysis of heavy metal content in Solanum nigrum tissue is ...

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Abstract

The invention discloses a plant repair method of applying a transgenetic plant composite system to polluted soil. A transgenetic plant is obtained by steps of construction of a plant transformation vector, plant genetic transformation and the like. The transgenetic plant can be used for plant repair of heavy metal polluted soil and water bodies. The transgenetic plant shows a hyperaccumulating characteristic and resistance to different heavy metals such as lead and cadmium, can be widely applied to repairing heavy metal polluted soil, and has efficient repair efficiency to the polluted soil. The plant repair method can be used to polluted soil caused by mining, soil pollution of surrounding earth piles, and polluted soil of a waste site of an electroplate factory and the periphery, for example, various composite metal polluted soil such as Cd, Cr, Cu, Pb, Z and Ni. The plant repair method has the multiple advantages of being high in adaptability, environmentally friendly and the like in repair of composite polluted soil, and has a wide application prospect.

Description

technical field [0001] The invention relates to the fields of biotechnology and soil restoration, in particular to a method for specific expression of exogenous genes in restoration plants and a heavy metal restoration technology applied to polluted soil. Background technique [0002] Soil is the natural environment on which human beings depend and an important resource for agricultural production. Various food, resource and environmental problems faced by the world are closely related to soil. Since the last century, the continuous development of mining and metallurgy, and the widespread use of various chemical products, pesticides and fertilizers have led to the emergence of heavy metal soil and groundwater pollution. Soil heavy metal pollution refers to the phenomenon that human activities bring heavy metals into the soil, resulting in changes in soil composition and physical and chemical properties, resulting in degradation of soil quality and ecological deterioration. ...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H4/00A01H5/00A01H6/82B09C1/10B09C1/00
CPCA01H4/001A01H4/008B09C1/10B09C1/105C12N15/8259
Inventor 卢宏玮卢静昭雷凯文王伟鹏康瑜
Owner INST OF GEOGRAPHICAL SCI & NATURAL RESOURCE RES CAS
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