Composition for phytochelatin transport

A technology of phytochelatin peptide and composition, which is applied in the field of compositions for transporting phytochelatin peptides, and can solve problems to be determined and the like

Inactive Publication Date: 2013-07-31
POSTECH ACAD IND FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the transporters involved in the transport of PC-bound heavy metals to plant vacuoles and those involved in the transport of PC-bound As(III) remain to be identified

Method used

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  • Composition for phytochelatin transport
  • Composition for phytochelatin transport
  • Composition for phytochelatin transport

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1: Sensitivity test of atabcc1 and atabcc2 double gene knockout mutants to arsenic, arsenic-containing herbicides or cadmium

[0074] The Arabidopsis ABCC gene was isolated from a total of 15 Arabidopsis homozygous knockout lines, and the mutants were grown in the presence of arsenic, arsenic-containing herbicides, and cadmium. Except kindly provided by Dr. Markus Klein (Frelet-Barrand A et al. (2008), Comparative mutant analysis of Arabidopsis ABCC-type ABC transporters: AtMRP2 contributes to detoxification, vacuolar organic anion transport and chlorophyll degradation. Plant Cell Physiol 49:557-569) All abcc knockout mutants except abcc2-2, abcc11-1 and abcc12-2 were obtained from the Salk Genome Analysis Laboratory (http: / / signal.salk.edu / ). The accession and knockout numbers of all 15 abcc mutants are listed in Table 1. Among other things, a new mutant allele, Salk_017431 of ABCC1, was used in this experiment, and its nomenclature was named abcc1-3 following...

Embodiment 2

[0079] Example 2: Examination of whether AtABCC1 and AtABCC2 improve arsenic resistance and accumulation in budding yeast in the presence of PCS1 (phytochelatin synthase 1)

[0080] To determine whether AtABCC1 and AtABCC2 are resistant to arsenic (As), the PCR products of yhl035c::HIS3-MX6, yll015w::Kan-MX6 and yll048c::TRP1-MX6 were inserted by homologous recombination into the Cd-sensitive mutant, DTY167 Homologous loci (YHL035c, YLL015w and YLL048c) of yeast gDNA and preparation of budding yeast SM4 mutants lacking YCF1 and three other vacuolar ABCC-type ABC transporters, YHL035c, YLL015w and YLL048c, in the preparation of SM4 Expression of the transporters AtABCC1 and AtABCC2.

[0081] To amplify DNA fragments for homologous recombination, the following primers were used with genomic DNA obtained from a single gene null mutant: For yhl035c::HIS3-MX6, the primers were His3MX6-YHL035F (SEQ ID NO: 7) and His3MX6- YHL035R (sequence number: 8), genomic DNA is obtained from YM...

Embodiment 3

[0092] Example 3: Whether AtABCC1 and AtABCC2 can be tested as phytochelatin transporters

[0093] With empty vector (pNEV-Ura) (Sauer N&Stolz J (1994) SUC1 and SUC2: two sucrose transporters from A. thaliana; expression and characterization in baker's yeast and identification of the histidine-tagged protein. Plant J 6:67-77) , pNEV-AtABCC1 or pYES3-AtABCC2 (Lu YP et al. (1998) AtMRP2, an Arabidopsis ATP-binding cassette transporter able to transport glutathione S-conjugates and chlorophyll catabolites: functional comparisons with AtMRP1. Plant Cell10: 267-282) transformed SM7 Yeast cells were grown in SD-uracil liquid medium (Qbiogene) at 30°C for 4 hours, and then centrifuged at 2,000 g for 15 minutes to collect cell micromass. All cell lines were cultured in YPD medium (USB, Duchefa) at 30°C for 30 minutes, centrifuged at 2,000 g for 15 minutes to collect cell micelles, which were then lysed with cytolytic enzyme (1,000 Unit / g cell fresh weight, Sigma). Microsomal vesicle...

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Abstract

The present invention relates to a composition for phytochelatin transport, comprising DNA molecules encoding ABCC (MRP, multidrug resistance-associated protein)-like ABC (ATP-binding cassette) transporter proteins in plants. The composition for phytochelatin transport according to the present invention can accumulate and sequester phytochelatin alone or in combination with a toxic substance by transporting same into the vacuole of a cell, and thus reduce the content of the toxic substance which moves from the root of a plant to the shoot thereof. Therefore, the DNA molecules of the present invention can be applied to the development of crops in which the content of toxic substances existing in edible plant parts such as leaves, stems, and fruits are reduced. Furthermore, the DNA molecules of the present invention are applied to inedible plants to develop plants which are useful in phytoremediation, or phytoextraction, by increasing the accumulated amounts of arsenic, arsenic compounds, and cadmium, and the resistances thereto, thereby economically cleaning the environment in an environmentally friendly manner is possible.

Description

technical field [0001] The present invention relates to compositions for transporting phytochelatins comprising DNA molecules encoding the ABCC (Multidrug Resistance-Associated Proteins (MRP)) subfamily of ATP-binding cassette (ABC) transporters in plants. Background technique [0002] Arsenic (As) has been widely used in medical, industrial and agricultural applications. In the field of medicine, arsenic (As) has been used in the treatment of diseases such as syphilis, trypanosomiasis or amoebic dysentery. In agricultural applications, arsenic-containing herbicides such as disodium methyl arsenate (DSMA) have been used to control weeds and harmful insects. However, arsenic (As) is a highly toxic environmental pollutant known to cause a variety of health-related problems. Arsenic (As) contamination of groundwater has been reported in some countries, such as China, Thailand, and the United States, where mining operations are active, as well as Bangladesh and India. Contami...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/87A01H5/00
CPCC12N15/8271C12N15/8259C07K14/705
Inventor 李永淑恩里科·马丁诺亚朴祉映宋元镛大卫·G·门多萨科扎特朱利安·I·施罗德
Owner POSTECH ACAD IND FOUND
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