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Arabidopsis thaliana nitrate transporter gene NRT3.1, and encoding protein and application thereof

A nitrate transport, Arabidopsis technology, applied in application, genetic engineering, plant genetic improvement and other directions

Inactive Publication Date: 2014-03-26
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the regulation of aboveground biomass by this gene under low nitrogen conditions.

Method used

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  • Arabidopsis thaliana nitrate transporter gene NRT3.1, and encoding protein and application thereof
  • Arabidopsis thaliana nitrate transporter gene NRT3.1, and encoding protein and application thereof
  • Arabidopsis thaliana nitrate transporter gene NRT3.1, and encoding protein and application thereof

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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 fields of plant molecular biology and transgenic correlation techniques, and particularly relates to verification of low-nitrogen-resistance function of Arabidopsis thaliana nitrate transporter gene NRT3.1 and application of the gene in culture of new species of low-nitrogen-resistance nitrogen nutrition high-efficiency plants. By using the model plant Arabidopsis thaliana as the material, the homozygous strain system of the NRT3.1 gene T-DNA insertion mutant CS876661 is utilized to research the functions of the NRT3.1 gene in the plant low-nitrogen high-efficiency aspect. The experimental result shows that the dry weight of the above-ground part of the NRT3.1 gene knockout mutant is obviously lower than that of the wild type (P=0.002) under low-nitrogen culture conditions (0.1mM NO3<->), and the nitrogen contents of the root and above-ground part are respectively obviously lower than those of the wild type (P=0.045, P=0.01), which indicates that the NRT3.1 gene has important functions in signal transmission paths under plant response low-nitrogen stress. The overexpression of the gene is estimated to enhance the low nitrogen resistance of the plant and has great economic value in plant nitrogen nutrition high-efficiency molecule 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 NRT3.1 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 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-efficiency...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415A01H5/00
Inventor 徐江徐小栋东方阳鲁黎明王西瑶张少斌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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