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Rice nitrate reductase gene nr2 and its encoded protein and application

A technology of reductase and protein, applied in application, genetic engineering, plant gene improvement, etc., can solve the problems of separation that have not been reported yet, and achieve the effect of improving utilization efficiency and increasing yield

Active Publication Date: 2011-11-30
CHINA NAT RICE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition to the cloned NADH-type nitrate reductase gene NR1 in rice, the isolation of the NR2 gene encoding NAD(P)H-specific nitrate reductase has not been reported yet.

Method used

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  • Rice nitrate reductase gene nr2 and its encoded protein and application
  • Rice nitrate reductase gene nr2 and its encoded protein and application
  • Rice nitrate reductase gene nr2 and its encoded protein and application

Examples

Experimental program
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Effect test

Embodiment 1

[0038] 1. Rice material

[0039] The japonica rice variety (Oryza sativa L.japonica) "Nipponbare" and the indica rice variety (Oryza sativa L.indica) "Guangluai 4".

[0040] 2. Potassium chlorate sensitivity detection for targeted populations

[0041] Using Guanglu Ai 4 and Nippon Clear's F 2 Population (720 lines) and recombinant inbred line F 7 The population (646 strains) was used as the targeting population. After the seeds germinate in an incubator at 28°C for 5 days (change the single distilled water once a day), take 10 seeds that germinate consistently from each strain, wrap half of them with germination paper and put them into a test tube filled with single distilled water As a control, the other half was wrapped with germination paper and put into a test tube equipped with 0.1% potassium chlorate solution as a treatment, and then put into a light incubator at 28°C for 12 hours of light and 12 hours of darkness. After cultivating for 7 days, the total length from ...

Embodiment 2

[0055] Plant Transformation and Functional Analysis:

[0056] The transformation vector utilizes the 9.776kb fragment (including the complete 3 introns and 4 exons of the 3.039kb NR2 gene, the complete 3 introns and 4 exons, 5.359 kb upstream sequence and 1.378kb downstream sequence) were cloned into the binary vector pCAMBIA1300 ( image 3 ). The plasmid was transferred into Agrobacterium tumefaciens strain EHA105 by electric shock method to transform rice. We used the callus induced by Nipponbare immature embryos, and after 3 weeks of culture in the induction medium, we selected the vigorously growing callus as the recipient of transformation. Using the conventional rice transgenic method, the rice callus was infected with the EHA105 strain containing the binary plasmid vector, co-cultivated in the dark at 25°C for 3 days, and cultured on a selection medium containing 40 mg / L hygromycin. Resistant calli were cultured for 10 days on pre-differentiation medium containing 50 ...

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Abstract

The invention discloses a rice nitrate reductase gene NR2. The sequence of the gene may be: (a) a genomic nucleotide sequence represented by Seq ID No.1; (b) a cDNA nucleotide sequence represented by Seq ID No.2; or (c) nucleotide sequence of a mutant gene, allele, derivative thereof or the like, which is formed by adding, substituting or losing one or several nucleotides in the nucleotide sequence (a) or the nucleotide sequence (b) and can code a protein having nitrate reductase activity. The invention also discloses a protein (with an amino acid sequence represented by Seq ID No.3) coded bythe gene and the use of the gene in regulation of absorption and utilization of nitrogen. Through the cloning and use of the NR2 gene, the absorption and use of nitrogen by rice can be effectively regulated, the efficiency of nitrogen utilization can be improved, and finally the yield of the rice can be increased.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to rice nitrate reductase gene NR2 and its encoded protein and application. Background technique [0002] As a monocot model plant, rice is the staple food of more than half of the population in my country, and it is also one of the food crops that consume the most nitrogen fertilizer. According to statistics, my country accounts for more than 1 / 3 of the world's nitrogen fertilizer consumption for rice. Nitrogen is the primary component of plant growth and the limiting factor for yield formation among all the essential nutrients of rice. Therefore, the research and improvement of nitrogen nutrition in rice can not only provide the necessary basis for the rational use of nitrogen fertilizer in rice cultivation, but also has potential agricultural production value. [0003] Nitrogen exists in various molecular forms, such as nitrate nitrogen, ammonium nitrogen, and urea. Ri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N9/06C12N15/63C12N1/21C12N5/10A01H5/00
Inventor 钱前高振宇滕胜郭龙彪
Owner CHINA NAT RICE RES INST
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