Arabidopis thaliana ammonium ion transporter gene AMT1.1 and coding protein and usage thereof

A kind of Arabidopsis ammonium, gene technology, applied in the application, genetic engineering, plant genetic improvement and other directions

Inactive Publication Date: 2013-09-04
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far there is no detailed report on the regulation of the above-ground growth and development of this gene under low-nitrogen conditions

Method used

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  • Arabidopis thaliana ammonium ion transporter gene AMT1.1 and coding protein and usage thereof
  • Arabidopis thaliana ammonium ion transporter gene AMT1.1 and coding protein and usage thereof
  • Arabidopis thaliana ammonium ion transporter gene AMT1.1 and coding protein and usage thereof

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Experimental program
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Embodiment Construction

[0015] The present invention will be further described below in conjunction with experimental methods and data.

[0016] 1. Arabidopsis seedling culture:

[0017] Wild-type and mutant seeds were sterilized by soaking in 0.5% sodium hypochlorite solution for 10 min, and the sterilized seeds were placed in a 4°C refrigerator for vernalization for 2 days, and then spread on MS (Murashige and Skoog) solid medium containing 1% agar. Germinate and grow in a light incubator at 22°C.

[0018] 2. Identification of homozygous mutants:

[0019] DNA extraction: All experimental drugs used were purchased from Zhongke Ruitai Bioengineering Co., Ltd. Take about 0.5g fresh leaves of Arabidopsis thaliana, grind them into a paste, add 400μl SDS extract (0.5% SDS (W / V); 200mM Tris-Hcl, PH=8.0; 250mM NaCl; 25mM EDTA); 12,000r / min, Centrifuge for 3 minutes; take the supernatant, add an equal volume of isopropanol to shake gently, let it stand for 30 minutes; Add 30 μl of water and store at 4°C...

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Abstract

The invention belongs to the field of technology related to plant molecular biology and transgenosis. The invention concretely relates to verification of Arabidopis thaliana ammonium ion transporter gene AMT1.1 low nitrogen resisting function, and the usage of the gene in the cultivation of low nitrogen resisting and nitrogen element nutrition high efficiency plant new species. The invention employs the plant Arabidopis thaliana as material, and is used for researching function of AMT1.1 gene in the aspect of plant low nitrogen high efficiency by inserting AMT1.1 gene T-DNA into a mutant salk_091322 homozygous lines. The experiment result displays that the shoot dry weight of mutant with AMT1.1 gene knocked out is substantially less than that of the wild type in the low nitrogen culture condition; the nitrogen contents of the shoot and root are significantly or even highly significantly lower than those of the wild type; the mutant appears a low nitrogen sensitive phenotype. The invention instructs that the AMT1.1 gene plays an important effect in the signal transduction pathway of plant response low nitrogen stress, and predicts that the excessive expression of the gene can improve the low nitrogen resisting capability of plant and has a larger economic value in the plant nitrogen element nutrition high efficiency molecular breeding.

Description

Technical field: [0001] The invention belongs to the related technical field of plant molecular biology and transgene. Specifically, the response and mechanism of the key gene AMT1.1 of the Arabidopsis ammonium transporters (Ammonium transporters, AMTs) family under low nitrogen stress, as well as the application of this gene in the cultivation of new plant varieties with high nutritional efficiency. Background technique: [0002] Nitrogen is one of the essential nutrients for plants, and is a component of proteins, nucleic acids, chlorophyll, enzymes, etc. in plants. Insufficient nitrogen supply will affect the growth and development of plants and limit the yield of crops; while excessive nitrogen fertilizer application will reduce nitrogen use efficiency, which not only causes waste of resources, but also causes environmental pollution. One of the important ways to solve this problem is to tap the genetic potential of plants, clone low-nitrogen tolerance and high-efficien...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415A01H5/00
Inventor 徐江徐小栋王西瑶张少斌东方阳鲁黎明迟志海洪雪麻艳超赵明丁在松付金东
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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