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Compositions for transporting phytochelatins

一种植物螯合肽、组合物的技术，应用在转运植物螯合肽的组合物领域，能够解决有待确定等问题

Inactive Publication Date: 2016-05-18
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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  • Compositions for transporting phytochelatins
  • Compositions for transporting phytochelatins
  • Compositions for transporting phytochelatins

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 for abcc2-2, abcc11-2 and abcc1 knockout genes kindly provided by Dr. Markus Klein (Frelet-Barrand A et al. (2008), Comparative mutant analysis of Arabidopsis ABCC-type ABC transporters: AtMRP2contributestodetoxification, vacuolarorganicaniontransportandchlorophylldegradation. PlantCellPhysiol49:557-569), Mutants were all 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 the previously ...

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] 用空载体(pNEV-Ura)(SauerN&StolzJ(1994)SUC1andSUC2:twosucrosetransportersfromA.thaliana;expressionandcharacterizationinbaker'syeastandidentificationofthehistidine-taggedprotein.PlantJ6:67-77)、pNEV-AtABCC1或pYES3-AtABCC2(LuYP等(1998)AtMRP2,anArabidopsisATP-bindingcassettetransporterabletotransportglutathioneS -conjugatesandchlorophyllcatabolites:functionalcomparisonswithAtMRP1.PlantCell10:267-282) Transformed SM7 yeast cells were grown in SD-uracil liquid medium (Qbiogene) at 30°C for 4 hours, and then centrifuged at 2,000g for 15 minutes to collect cell microparticles. group. 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 units / g cell fresh weight, Sigma). Microsomal vesicles were then isolated from the cell extracts using the method di...

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Abstract

The present invention relates to a composition for transporting phytochelatins comprising two DNA molecules encoding two members of the ABCC (multidrug resistance-associated protein (MRP)) subfamily of ATP-binding cassette (ABC) transporters in plants. By transporting phytochelatins and / or noxious substances into cell vacuoles, the composition for transporting phytochelatins can be used alone or in combination with noxious substances to accumulate and sequester phytochelates, thereby reducing toxic substances in the cytoplasm. It also reduces the content of toxic substances that migrate from the roots to the stems of plants. Thus, said DNA molecules can be used to develop crops in which the content of toxic substances present in the edible parts of the plant, such as leaves, stems, fruits, is reduced. Furthermore, the DNA molecules can be used to develop inedible plants for phytoremediation or phytoextraction, and to increase the accumulation and resistance of plants to arsenic, arsenic compounds, and cadmium, possibly in an environmentally friendly and economically clean manner Phytoremediation or Phytoextraction.

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 Patents(China)
IPC IPC(8): C12N15/29C12N15/87A01H5/00
CPCC07K14/705C12N15/8259C12N15/8271
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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