Cultivation method of transgenic composite type ipomoea aquatica forsk for removing water body organic pollutants

A technology of organic pollutants and cultivation method, which is applied in the field of cultivation of transgenic compound water spinach, can solve problems such as no research reports on repairing organic pollutants, and achieves the effect of improving comprehensive repairing ability and improving repairing ability.

Inactive Publication Date: 2015-01-28
HANGZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The remediation effect of water spinach mentioned above is limited to the remediation of N...

Method used

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  • Cultivation method of transgenic composite type ipomoea aquatica forsk for removing water body organic pollutants
  • Cultivation method of transgenic composite type ipomoea aquatica forsk for removing water body organic pollutants
  • Cultivation method of transgenic composite type ipomoea aquatica forsk for removing water body organic pollutants

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 water spinach tissue culture rapid propagation aseptic seedling

[0028] (1) The seeds of different genotypes such as farm water spinach, willow-leaf water spinach, broad-leaf water spinach, and Thai water spinach were selected as experimental starting materials.

[0029] (2) Select different disinfection methods to sterilize the seeds. The seed pollution situation after disinfection is shown in Table 1. 0.1% mercuric chloride refers to a mass concentration of 0.1% mercuric chloride aqueous solution, and 75% alcohol is a volume concentration of 75% ethanol aqueous solution. The % sodium hypochlorite is 20% sodium hypochlorite aqueous solution. The results show that the disinfection effect of 0.1% mercuric chloride for 7.5 minutes is the best for seeds without pollution, and the germination rate of willow-leaf water spinach is the highest.

[0030] Table 1 Seed disinfection

[0031]

[0032] (3) Select different culture media and perform tissue culture o...

Embodiment 2

[0037] The acquisition method of embodiment 2 trans cyp2e1 gene water spinach

[0038] (1) Preparation of Agrobacterium rhizogenes infection solution containing cyp2e1 gene

[0039] The cyp2e1 gene is cloned from rabbit hepatocytes by RT-PCR. The GenBank accession number of the cyp2e1 gene is M15061, and the nucleotide sequence is shown in SEQ ID NO:1.

[0040]

[0041]

[0042] The plant transgene expression vector pSLD50-6 containing cyp2e1 gene was provided by Professor Doty of Washington University.

[0043] Using the freeze-thaw method, pSLD50-6 was introduced into competent Agrobacterium rhizogenes K599 cells (purchased from the National Collection of Plant Pathogenic Bacteria in U.K.), and recombinant Agrobacterium rhizogenes K599 / pSLD50-6 was constructed.

[0044] Inoculate recombinant Agrobacterium rhizogenes K599 / pSLD50-6 in LB liquid medium containing Km (kanamycin) 50 mg / L + Str (streptomycin) 50 mg / L, and cultivate to OD at 28°C and 200 r / min 600 0.4-0.6, ...

Embodiment 3

[0058] Example 3 Water spinach containing cyp2e1 gene's resistance and absorption analysis to water organic pollutants

[0059] The water spinach plant containing the cyp2e1 gene prepared in Example 2 (the transgenic plant obtained by growing hairy roots at the position of stem height 4-5cm) and the control plant (wild type) were respectively treated with 10 μg / mL benzene 1 / 2MS culture medium or 1 / 2MS culture medium containing 15μg / mL trichlorethylene, culture temperature is 18-25℃. The results showed that after 7 days of culture, the control non-transgenic plants withered in 1 / 2MS medium containing 10 μg / mL benzene or 1 / 2MS medium containing 15 μg / mL trichlorethylene, while the water spinach plants containing cyp2e1 gene remained normal growth ( Figure 5 ). With reference to methods such as Doty (Doty SL, Shang QT, Wilson AM, Westergreen AD, Newman LA, Strand SE, Gordon MP.Enhanced metabolism of halogenated hydrocarbons in transgenic plants containing mammalian cytochrome ...

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Abstract

The invention discloses a cultivation method of transgenic ipomoea aquatica forsk for removing water body organic pollutants; firstly, a plasmid vector containing a cyp2e1 gene is transferred into agrobacterium rhizogenes, a hairy adventitious root transferred with the cyp2e1 gene is obtained in a living plant by utilizing an agrobacterium rhizogenes transgenic method, and then the composite type transgenic ipomoea aquatica forsk containing the cyp2e1 gene hairy root is obtained; and the water body organic pollutants are absorbed by using the composite type transgenic plant.

Description

(1) Technical field [0001] The invention relates to a method for removing organic pollutants in water bodies, in particular to a method for cultivating transgenic composite water spinach for removing organic pollutants in water bodies. (2) Background technology [0002] Water spinach (Ipomoea aquatica Forsk) belongs to Convolvulaceae herbaceous plants, commonly known as bamboo leaf vegetables, there are two ecological types of aquatic and xerophytic. Water spinach can be planted once and harvested multiple times, so that nutrients can be directly removed from polluted water without secondary pollution. [0003] Some scholars have studied the function of water spinach in water purification. Li Yuru and others planted water spinach on heavily polluted rivers in Suzhou to control water pollution. Not only has the static test been carried out, but also a demonstration application has been made on the Miaojia River, a heavily polluted river course. The static test results show...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H4/00A01H5/00C02F3/32C02F101/38C02F101/36
Inventor 孙扬向太和韩依萱王嫚宋亚玲吴玲
Owner HANGZHOU NORMAL UNIVERSITY
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