Arabidopsis thaliana nitrate transporter gene NRT2.1, encoding protein and applications thereof

A technology of nitrate transport and Arabidopsis, applied in application, genetic engineering, plant genetic improvement, etc.

Inactive Publication Date: 2013-08-21
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Arabidopsis thaliana nitrate transporter gene NRT2.1, encoding protein and applications thereof
  • Arabidopsis thaliana nitrate transporter gene NRT2.1, encoding protein and applications thereof
  • Arabidopsis thaliana nitrate transporter gene NRT2.1, encoding protein and applications thereof

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Embodiment Construction

[0013] The present invention will be further described below in conjunction with experimental methods and data.

[0014] 1. Arabidopsis seedling culture:

[0015] 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, placed on Germinate and grow in a light incubator at 22°C.

[0016] 2. Identification of homozygous mutants:

[0017] 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 st...

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Abstract

The invention belongs to the plant molecule biology and transgenic related technical field, and specifically relates to verification of low nitrogen resistance of arabidopsis thaliana nitrate transporter gene NRT2.1 and applications of the arabidopsis thaliana nitrate transporter gene NRT2.1 in breeding of new low nitrogen resistance and nitrogen nutrition efficiency plant varieties. According to the present invention, a model plant arabidopsis thaliana is adopted as a material, and NRT2.1 gene T-DNA is adopted to insert into a mutant salk_008278 homozygosis line so as to research functions of the NRT2.1 gene in the field of plant low nitrogen efficiency, wherein experiment results show that lateral roots of the NRT2.1 knocked out mutant is significantly more than lateral roots of the wild-type (P=0.044) under a high sucrose/low nitrogen (7.5% sucrose, 0.1 mM NH4NO3) culture condition, such that the NRT2.1 gene provides important effects in a signal transduction pathway of the plant response to low nitrogen stress, and it can be predicted that overexpression of the gene can increase low nitrogen resistance of the plant and can provide high economic values in plant nitrogen nutrition 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 NRT2.1 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...

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Application Information

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IPC IPC(8): C12N15/29C07K14/415A01H5/00
Inventor 徐江徐小栋张少斌东方阳鲁黎明王西瑶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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