Method for improving degradation ability of transgenic plants to polycyclic aromatic hydrocarbons

A technology of transgenic plants and polycyclic aromatic hydrocarbons, applied in the fields of botany equipment and methods, biochemical equipment and methods, plant products, etc., can solve the problems of weak hydroxylation of polycyclic aromatic hydrocarbons and low degradation ability of polycyclic aromatic hydrocarbons, and achieve Effects of less environmental impact, improved tolerance and degradability

Inactive Publication Date: 2013-12-25
上海瑞丰农业科技有限公司 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the weak hydroxylation of PAH...

Method used

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  • Method for improving degradation ability of transgenic plants to polycyclic aromatic hydrocarbons
  • Method for improving degradation ability of transgenic plants to polycyclic aromatic hydrocarbons
  • Method for improving degradation ability of transgenic plants to polycyclic aromatic hydrocarbons

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1, Effects of Transgenic Arabidopsis on Phenanthrene and Pyrene Tolerance and Degradability

[0063] 1) Preparation of Agrobacterium Competent Cells

[0064] The Agrobacterium strain is Agrobacterium tumefaciens EHA105, or LBA4404, or GV3101 strain (Biovector Co., LTD). Pick a single bacterium and culture it overnight in 25ml YEB medium (added with 50mg / l rifampicin), transfer 5ml of the bacterial liquid to 100ml YEB medium (added with 50mg / l rifampicin), and cultivate to OD600=0.7-0.8, Place the bacterial solution on ice for 10 minutes, centrifuge at 5000rpm for 10min, 4°C, collect the bacterial cells, add 100ml sterile double distilled water to wash twice. Add 4ml of 10% glycerol to suspend the bacteria and transfer to a 50ml centrifuge tube. Centrifuge at 5500rpm for 10min at 4°C. Collect the bacteria, add 500μl 10% glycerol to suspend the bacteria, and transfer to a 1.5ml centrifuge tube.

[0065] 2) The bivalent gene plant expression vector pCYPGST was ...

Embodiment 2

[0078] Embodiment 2, the impact of transgenic rice on the degradation ability of phenanthrene and pyrene

[0079] 1) Agrobacterium-mediated transformation of rice

[0080] The rice callus was transformed with the Agrobacterium strain containing the expression vector pCYPGST obtained in step 2) of Example 1. N6 medium is the basic medium, shelled rice seeds, and immature embryos 12-15 days after pollination are surface-sterilized and inoculated into N6D2 medium to induce callus (N6 medium, hydrolyzed milk protein 500mg / L, Sucrose 30g / L, 2,4-D2mg / L, Phytogel 2.5g / L, pH 5.8); after 4-7 days of culture, callus was taken for transformation. After the Agrobacterium containing the expression vector pCYPGST was cultured at OD0.8-1.0, centrifuge at 5000g for 8 minutes, wash once with reconstituted water, suspend and infect with an equal volume of MS culture medium for 8 minutes, and blot dry the culture medium placed in MSO+NAA1+BA2 in 22°C for 3 days. Then transfer to selection med...

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Abstract

The invention discloses a method for improving degradation ability of transgenic plants to polycyclic aromatic hydrocarbons. The method comprises the following specific steps of constructing a divalent gene plant expression vector by using a transformed P450 mono-oxygenase gene and a synthesized grape glutathione s-transferase (GST) gene, and then converting the divalent gene expression vector to plants through agrobacterium tumefaciens mediated transformation. The tolerance and the degradation ability of the transgenic plants obtained by using the method to the polycyclic aromatic hydrocarbons are increased; and the planting of the transgenic plants is beneficial to the remediation of soil environments polluted by the polycyclic aromatic hydrocarbons.

Description

technical field [0001] The invention belongs to the field of environmental science, and in particular relates to a method for improving the ability to degrade polycyclic aromatic hydrocarbons by utilizing transgenic plants. Background technique [0002] Polycyclic aromatic hydrocarbons (PAHs) and their derivatives are a class of organic pollutants widely present in the environment. Polycyclic aromatic hydrocarbons are hydrocarbons containing two or more benzene rings, which can be divided into aromatic fused ring and non-aromatic fused ring types. In environmental pollution research, polycyclic aromatic hydrocarbons generally refer to aromatic fused ring types, of which more than 16 Aromatic compounds have been included in the US Environmental Protection Agency's blacklist of priority pollutants. Polycyclic aromatic hydrocarbons are mostly produced by fossil fuels such as petroleum and coal, and hydrocarbon-containing substances such as wood, natural gas, organic polymer co...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H5/00
Inventor 彭日荷姚泉洪王荣谈付晓燕田永生赵伟严培兰臧晓韵王波王丽娟
Owner 上海瑞丰农业科技有限公司
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