Method for improving plant absorption and tolerance methanal, plant expression vector and application

A plant expression vector, plant absorption technology, applied in the directions of botanical equipment and methods, angiosperms/flowering plants, biochemical equipment and methods, etc., can solve problems such as improving the absorption of formaldehyde by indoor plants, and achieve the speed of absorbing formaldehyde. good growth effect

Inactive Publication Date: 2008-09-24
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] There are no plants transfected with DAS or DAK genes, and no other gene manipulations have been reported to improve the absorption of formaldehyde in indoor plants.

Method used

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  • Method for improving plant absorption and tolerance methanal, plant expression vector and application
  • Method for improving plant absorption and tolerance methanal, plant expression vector and application
  • Method for improving plant absorption and tolerance methanal, plant expression vector and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0132] Example 1, Construction of DAS Gene Recombinant Plant Expression Vector pK2-35S-PrbcS-*T-DAS

[0133] (1) Preparation and detection of Candida boidinii genomic DNA

[0134] The Candida boidinii used in the present invention was purchased from China Industrial Microorganism Culture Collection Center, and the Candida boidinii genomic DNA was prepared by the CTAB method. Take 1.5mL bacterial liquid and centrifuge at 4°C 4000rpm for 2min, discard the supernatant, and collect the bacterial cells. Add 400 μl 2×CTAB (Tris-HCl pH 7.5 100 mM, EDTA 20 mM, NaCl 1.4 M, CTAB 2%) to homogenate, incubate at 65° C. for 20 minutes, add 500 μl chloroform to mix, and centrifuge at room temperature at 13,000 rpm for 10 minutes. Transfer the supernatant, add 50 μl of 10% CTAB, add 650 μl of isopropanol and place at room temperature for 1 hour, centrifuge at 12,000 rpm at 4°C for 25 minutes, wash the precipitate once with 500 μl of 75% alcohol, dry it in vacuum, add 20 μl of TE containing R...

Embodiment 2

[0145] Example 2, Construction of DAK Gene Recombination Vector pH2-35S-PrbcS-*T-DAK

[0146] (1) Preparation and detection of Pichia pastoris genomic DNA

[0147] Pichia pastoris (Pichia pastoris) was purchased from China Industrial Microorganism Collection Center, and the genomic DNA of Pichia pastoris was prepared by the CTAB method. Take 1.5mL of the bacterial liquid and centrifuge at 4000rpm at 4°C for 2min, discard the supernatant, and collect the bacterial cells. Add 400 μl 2×CTAB (Tris-HCl pH 7.5 100 mM, EDTA 20 mM, NaCl 1.4 M, CTAB 2%) to homogenate, incubate at 65° C. for 20 minutes, add 500 μl chloroform to mix, and centrifuge at room temperature at 13,000 rpm for 10 minutes. Transfer the supernatant, add 50 μl of 10% CTAB, add 650 μl of isopropanol and place at room temperature for 1 hour, centrifuge at 12,000 rpm at 4°C for 25 min., wash the pellet once with 500 μl of 75% alcohol, dry it in vacuum, add 20 μl of RNase-containing TE to dissolve, 37 °C for one hour...

Embodiment 3

[0158] Embodiment 3, transform Agrobacterium with the plant expression vector that contains DAS and DAK gene

[0159] Competent cells of Agrobacterium were prepared, and the above constructed plant expression vectors pK2-35S-PrbcS-*T-DAS and pH2-35S-PrbcS-*T-DAK were transferred into Agrobacterium (C58Cl(pPMP90) ), transformants were selected on plates added with spectinomycin. Take a small amount of plasmid and add it to the competent cells of Agrobacterium, mix gently; add the mixture into the pre-cooled electroporation cup, tap the cup body gently to make the mixture fall to the bottom of the cup; place the electroporation cup on the electroporation In the chute of the instrument (BIO-RAD), use a 1mm electric shock cup and 200 ohm, 2.5kV / 0.2cm parameters for electric shock, immediately take out the electric conversion cup after electric shock, quickly add 0.5ml SOC medium, mix well, transfer to 1.5 ml centrifuge tube; 28°C, 200rpm shaker culture 3-5h; room temperature, 750...

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Abstract

The invention discloses a method for improving assimilation of formaldehyde and tolerance to formaldehyde of a plant, expression vectors and application of the plant, which belongs to the technical field of plant gene engineering. In the method, double genes containing DAS and DAK are fed into the chloroplasts of plant leaf cells, thus establishing a formaldehyde assimilation path, and a mechanism to assimilate formaldehyde with the photosynthesis of the plant so as to detoxicate formaldehyde in plant cells, create decorative plants with the specific function of decontaminating formaldehyde in indoor air and controlling the pollution of formaldehyde in indoor air, and maintain health of human bodies. The expression vectors of the plant are vector pK2-35S-PrbcS-*T-DAS and vector pH2-35S-PrbcS-*T-DAK. Compared with the control plant, the plant can thrust up in the environment polluted by formaldehyde, the quantity of formaldehyde assimilated is increased by 20 to 70 percent, and the speed to assimilate formaldehyde is improved by 16 to 27 percent.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and specifically relates to a method for improving plant absorption and tolerance to formaldehyde, a plant expression vector required by the method, a construction method thereof, and applications thereof. Background technique [0002] Formaldehyde (HCHO) is widely used in industrial production as a raw material for the manufacture of resins, adhesives, paints, plastics, and man-made fibers, and is the most widely used chemical raw material in the adhesive industry. With the development of the economy and the improvement of people's living standards, various building decoration materials made of raw materials containing formaldehyde have entered the family. People move into new houses to carry out interior decoration and update furniture, which is the main source of indoor air formaldehyde pollution, making formaldehyde a ubiquitous air polluting gas. Long-term exposure to low-dose forma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/31A01H1/00A01H5/00
Inventor 陈丽梅李昆志潘正波宋中邦赵艳
Owner KUNMING UNIV OF SCI & TECH
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