Rice high affinity nitrate transport protein gene OsNAR2.1

A nitrate and high affinity technology, applied in the field of genetic engineering, can solve the problems of low nitrogen utilization rate and nitrogen loss

Active Publication Date: 2009-04-01
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are a series of environmental problems caused by low nitrogen use efficiency and a large amount of nitrogen loss in China.

Method used

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  • Rice high affinity nitrate transport protein gene OsNAR2.1
  • Rice high affinity nitrate transport protein gene OsNAR2.1
  • Rice high affinity nitrate transport protein gene OsNAR2.1

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Experimental program
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Effect test

Embodiment Construction

[0021] specific implementation plan

[0022] 1. Cloning of rice high-affinity nitrate transport protein gene OsNAR2.1 sequence

[0023] 1) Extraction of total RNA. Rice (Nipponbare) seedlings grow to the 3-leaf stage. After 6 hours of treatment with 0.2mM nitrate nitrogen, the roots are immediately taken out and quickly stored in liquid nitrogen. Weigh about 0.1g of the roots and store them in liquid nitrogen. Grinding, grinding and fully adding to a 1.5ml centrifuge tube, quickly adding 1ml Trizol reagent (purchased from Invitrogen, USA), shaking well, and then extracting total RNA.

[0024] 2) Cloning of the full length of the OsNAR2.1 gene

[0025] The rice NAR2.1 gene series OsNAR2.1 (AP004023 .2), which is located on the second chromosome. Primers (below) were designed to fish out the full-length sequence of OsNAR2.1 from the tissue cDNA library. See SEQ ID NO.1 for the sequence.

[0026] P1:5'-CAATGGCGAGGCTAGCCGGCGTT-3'

[0027] P2:5'-CGATCTACTTGTCCTTCTTGCGCTTCT-3'...

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Abstract

The invention discloses a sequence of rice high-affinity nitrate transport protein gene OsNAR2.1 and application thereof, belonging to the gene engineering field. The invention also discloses a cDNA sequence SEQ ID NO.1 of the rice high-affinity nitrate transport protein gene OsNAR2.1 and a coded mino acid sequence SEQ ID NO.2 thereof. The gene function is firstly reported in rice and expressed in frogspawn heterogenous system to determine that the OsNAR2.1 protein can remarkably promote the transportation of nitrate by the rice; and mRNA expression analysis shows that the gene is subject to the induction of low nitrate nitrogen and the inhibition of ammonium and nitrogen metabolin. The knockout mutant of the gene causes the rice to reduce the absorption of nitrate and have obvious anazotic symptoms, the gene also influences the ripening rate of the rice through the field experiments, and therefore, over expression OsNAR2.1 can improve the absorption of nitrate nitrogen of the rice under low-nitrogen condition and the yield of the rice.

Description

technical field [0001] The invention discloses a rice high-affinity nitrate transport protein gene OsNAR2.1 sequence and its application, belonging to the field of genetic engineering. Specifically, the application of the gene significantly promotes the absorption of nitrate by rice. Background technique [0002] Nitrogen is one of the important macronutrients for crops and participates in various metabolic processes of organisms. It is a component of many living substances in plants, such as: amino acids, proteins, nucleic acids, enzymes, chlorophyll, etc. Nitrogen accounts for 1.5-2% of plant dry weight and 16% of total plant protein (Frink CR., Waggoner PE. and Ausubel JH. Nitrogen fertilizer: retrospect and prospect. Proc. Natl Acad. Sci. USA. 1999.96: 1175- 1180.). [0003] At present, China's nitrogen fertilizer use accounts for 30% of the global nitrogen fertilizer use (Peng Shaobing, Huang Jianliang, Zhong Xuhua, Yang Jianchang, Wang Guanghuo, Zou Yingbin, Zhang Fu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415A01H1/00A01H5/00
Inventor 严明范晓荣徐国华沈其荣
Owner NANJING AGRICULTURAL UNIVERSITY
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