Application of nitrogen transport gene OsNPF7.4 to rice selecting breeding

A rice and gene technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve problems affecting plant growth and development, and what type of plant growth and development is affected, so as to enhance tillering ability, increase the number of tillers, and increase plant yield. Effect

Active Publication Date: 2017-07-14
WUHAN INST OF BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although it is known that nitrogen nutrition can promote plant growth and development, there is no systematic understanding of what type of plant growth and development nitrogen nutrition affects.
In addition, the rice NPF family has more than 80 members. It is almost unknown which member of the NPF gene family responds to nitrogen nutrition, and at what site mediates nitrogen nutrition transport, thus affecting the growth and development of the plant.

Method used

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  • Application of nitrogen transport gene OsNPF7.4 to rice selecting breeding
  • Application of nitrogen transport gene OsNPF7.4 to rice selecting breeding
  • Application of nitrogen transport gene OsNPF7.4 to rice selecting breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 OsNPF7.4 Construction of Gene Interference Plants

[0022] Extract the RNA of rice Zhonghua 11 and reverse transcribe it into cDNA, using the primer pair:

[0023] F1: 5'- GGTACC ACGCCATGGAGAGAGGCCAGCA-3'( Kpn I),

[0024] R1: 5'- GGATCC AGGTGACCAGCACCATGCCCA-3'( Bam H I);

[0025] F2: 5'- ACTAGT ACGCCATGGAGAGAGGCCAGCA-3'( Speech I),

[0026] R2: 5'- GAGCTC AGGTGACCAGCACCATGCCCA-3'( Sac I);

[0027] PCR amplified OsNPF7.4 The cDNA fragment of the gene was digested by the corresponding restriction endonuclease and then connected into the pTCK303 vector to construct the OsNPF7.4 gene interference expression vector OsNPF7.4 - pTCK303. Using the genetic transformation method mediated by Agrobacterium EHA105, the interference expression vector was introduced into the normal japonica rice variety Zhonghua 11.

[0028] Transplant all the obtained transgenic seedlings into baskets with soil, water and fertilize them regularly, and plant them in...

Embodiment 2

[0035] Example 2 OsNPF7.4 Construction of Gene Overexpression Plants

[0036] Extract the RNA of rice Zhonghua 11 and reverse transcribe it into cDNA, using the primer pair:

[0037] F5: 5'- GGTACC ATGGACGCCGGCGACGCCATGGAG-3'( kpn I),

[0038] R5: 5'- TCTAGA TGACACGACCGTCTTCACCTTGTA-3'( Xba I);

[0039] Amplified by PCR OsNPF7.4 After cDNA of the gene, passed kpn I and Xba I was connected to the pCAMBIA-1306 vector (the pCAMBIA-1306 vector was purchased from Cambia Company), and the OsNPF7.4 gene overexpression vector OsNPF7.4 -p1306. Agrobacterium EHA105 Mediated genetic transformation method, the overexpression vector was introduced into the normal rice variety Zhonghua 11.

[0040] Transplant all the obtained transgenic seedlings into baskets with soil, water and fertilize them regularly, and plant them in the field when the seedlings grow about 10cm in height. After the seedlings grow up, extract genomic DNA and detect the transgenic plants by PCR ...

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Abstract

The invention discloses application of nitrogen transport gene OsNPF7.4 to rice selecting breeding and belongs to the field of plant genetic engineering. An amino acid sequence of an OsNPF7.4 gene coding protein is shown as SEQ ID NO.1, and a cDNA sequence is shown as SEQ ID NO.2. Normal rice tiller number, spike number and grouting grain number can be increased by reducing OsNPF7.4 gene expression after a rice OsNPF7.4 gene interfering plant and an OsNPF7.4 gene over-expression plant are built, therefore, the OsNPF7.4 gene can be applied to rice selecting breeding to improve rice yield. The OsNPF7.4 gene has important application value in the aspect of elaborating the influence of nitrogen transport to plant growing and developing processes.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, in particular to nitrogen transport genes OsNPF7.4 Application in rice breeding. Background technique [0002] Plants obtain nitrogen by absorbing ammonia, nitrate, amino acids, soluble peptides, etc. in the soil; nitrogen absorption and transport mainly rely on ammonium transport protein (AMT), nitrate transport protein (NRT), amino acid transport protein (AAT) , Peptide Transporter (PTR) and other transport proteins (Williams L, Miller A. Transporters responsible for the uptake and partitioning of nitrogenous solutes. Annual Review of Plant Physiology and Plant Molecular Biology, 2001, 52: 659-688.). Ammonium is absorbed by plant AMT to synthesize glutamine and glutamate by glutamine synthase (GS) and glutamate synthase (GOGAT), which in turn form other amino acids (Sonoda Y, Ikeda A, Saiki S , et al. Feedback regulation of the ammonium transporter gene family AMT1 by glutamine in ric...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/29C07K14/415A01H5/00
CPCC07K14/415C12N15/8218C12N15/8261
Inventor 方中明黄玮婷聂海鹏钱焌杰吕凯汪杰
Owner WUHAN INST OF BIOENG
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