Nitrate transporter gene AtNRT1.5, and coding protein and application thereof

A technology of nitrate transport and atnrt1.5, which is used in applications, genetic engineering, plant genetic improvement, etc.

Inactive Publication Date: 2013-12-11
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

But so far there is no report that this gene regulates plant growth and development under different nitrogen conditions.

Method used

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  • Nitrate transporter gene AtNRT1.5, and coding protein and application thereof
  • Nitrate transporter gene AtNRT1.5, and coding protein and application thereof
  • Nitrate transporter gene AtNRT1.5, and coding protein and application thereof

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Embodiment Construction

[0016] The present invention will be further described below in conjunction with experimental methods and data.

[0017] 1. Arabidopsis seedling culture:

[0018] 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.

[0019] 2. Identification of Homozygous Mutants

[0020] 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 plant molecular biology, and specifically relates to functional verification of arabidopsis thaliana nitrate transporter gene AtNRT1.5. A function of the gene in efficient utilization of plant nitrogen is studied by inserting T-DNA of the AtNRT1.5 gene into a homozygous line of a mutant CS870797. Experimental results show that the growth of the mutant on a high-nitrogen (MS) medium is inhibited; the root system of the mutant is obviously shorter than that of the wild type; and in a low nitrogen condition, the growths of the root systems of the mutant and the wild type have no difference, but dry weight of the mutant is lower than that of the wild type. Determination results of nitrogen content show that the nitrogen content in the root system of the mutant is higher than that of the wild type while the nitrogen content in the shoot is significantly lower than that of the wild type; and the difference is more obvious on the high nitrogen (MS) medium. The results demonstrate that after mutation of the ATNRT1.5 gene, nitrogen uptake capacity of the root system is not affected significantly while transportation capacity of the nitrogen from roots to shoots is reduced significantly. Over-expression of the gene is expected to improve the nitrogen utilization capability of plants and has relatively large economic value in plant nitrogen nutrition efficient 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 ATNRT1.5 of the Arabidopsis nitrate transporters (Nitrate transporters, NRTs) family under low nitrogen stress, as well as the application of this gene in the cultivation of new varieties of nutritionally efficient plants. 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-efficiency nitr...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415
Inventor 徐江徐小栋东方阳鲁黎明王西瑶张少斌麻艳超迟志海洪雪付金东赵明丁在松
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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