Gene for restoring cadmium contaminated soil and improving cadmium tolerance of plants and application of gene

A cadmium-contaminated soil and plant technology, applied in plant gene improvement, application, plant peptides, etc., can solve problems such as production reduction, crop pollution, and impact on sustainable development of agriculture and human society

Active Publication Date: 2017-11-07
HEFEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

All in all, cadmium easily enters the human body through food chains such as crops, which seriously threatens human health. At the same time, cadmium pollution also causes large-scale pollution of crops and reduces production, which affects the sustainable development of agriculture and human society.

Method used

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  • Gene for restoring cadmium contaminated soil and improving cadmium tolerance of plants and application of gene
  • Gene for restoring cadmium contaminated soil and improving cadmium tolerance of plants and application of gene
  • Gene for restoring cadmium contaminated soil and improving cadmium tolerance of plants and application of gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] MNB1 Acquisition of its coding gene

[0037] About 50-100 mg of wild-type Colombian ecotype Arabidopsis seedlings were used as materials, the total RNA was extracted with Trizol, and the RNA concentration was detected with an ultraviolet spectrophotometer. According to the instructions of the RevertAidTM First Strand cDNA Synthesis Kit (Fermentas Company), the first strand of cDNA was synthesized using the extracted total RNA as a template. Using the extracted first-strand cDNA as a template, carry out the following PCR reaction: 20 μl reaction system, containing 2 μl of 10×PCR buffer, 0.4 μl of dNTPs (10mM) mixture, 2 μl each of primer 1 and primer 2, and Tag enzyme (5U / μl) 0.2 μl, and add double distilled water to the rest to 20 μl. in,

[0038] Primer 1: F 5' NNNGGTACCATGTCTCGATTTGCTATCTTAGTAA3';

[0039] Primer 2: R 5' NNNGAATTCCTAATATTTGATGTAAGCAACAGAA3'.

[0040] Amplify on the Life Express gene thermal cycler: first pre-denaturation at 94°C for 5 min, then ...

Embodiment 2

[0042] Breeding cadmium-tolerant Arabidopsis

[0043] 1, MNB1 Gene Overexpression Transgenic Lines MNB1-OE1, OE2 the acquisition

[0044] To further verify the function of this gene in the regulation of heavy metal cadmium stress in plants, we constructed MNB1 Gene overexpression vector (35S: MNB1 ). Firstly, the target fragment is amplified. The wild-type Arabidopsis was normally cultured on ½MS medium for two weeks, the total RNA of the plant was extracted, reverse-transcribed to synthesize cDNA, and the synthesized cDNA was used as a template for PCR to amplify a sufficient amount of the target product, see figure 1 a. Using the PCR product as a template, the second amplification was carried out for the purpose of introducing enzyme cutting sites. The PCR product and the vector pCAMBIA1301 were digested and recovered, and then the recovered and purified target DNA fragment and the vector were ligated overnight with T4 DNA ligase. Transfer the above connection s...

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Abstract

The invention relates to a gene for improving cadmium tolerance of plants and application of an encoded protein of the gene in restoration of cadmium contaminated soil. The gene for restoring cadmium contaminated soil and improving cadmium tolerance of plants has a sequence shown as SEQ ID No:1 in a sequence table. The gene for improving cadmium tolerance of plants in the invention is transferred into the plants, so that the gene is excessively expressed in the plants, and the plant is cadmium tolerant. When the gene for improving cadmium tolerance of plants is excessively expressed, due to cadmium treatment, the gene can be combined with mannose, and increased expression of related genes such as GSH1, GSH2, PC1, PC2 and the like is promoted, so that the content of GSH and PC is increased. Outside cadmium ions are stored in vacuoles, tolerance to cadmium accumulation is further realized, and the cadmium contaminated soil can be restored to a certain degree.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and specifically relates to an application method for enhancing plant tolerance to cadmium poisoning by using genes and repairing cadmium-contaminated soil. Background technique [0002] With the development of industry and agriculture, soil cadmium pollution is becoming more and more serious, which seriously threatens human health. Heavy metal pollution has become one of the global environmental problems and one of the important problems facing the world today. Since cadmium is the most toxic heavy metal element in the environment, it has strong chemical activity, high mobility, and long-lasting toxicity in the environment. They cannot be degraded by microorganisms in water bodies, but can only be transformed and dispersed in various forms and enriched. It is absorbed and accumulated by the human body through the food chain, endangers human health, has carcinogenic, teratogenic, and mutag...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29A01H5/00
CPCC07K14/415C12N15/8271
Inventor 曹树青黄莹严星星张乘樊婷婷
Owner HEFEI UNIV OF TECH
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