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Application of Nitrogen Transport Gene osnpf8.1 in Improving Rice Tiller Number

A technology of tillering number and gene, applied in application, genetic engineering, plant genetic improvement and other directions, to achieve the effect of enhancing tillering ability

Active Publication Date: 2020-05-29
WUHAN INST OF BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are few reports about the research of NPF family members at present, and the present invention relates to OsNPF8.1 gene is a gene of rice NPF gene family

Method used

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  • Application of Nitrogen Transport Gene osnpf8.1 in Improving Rice Tiller Number
  • Application of Nitrogen Transport Gene osnpf8.1 in Improving Rice Tiller Number
  • Application of Nitrogen Transport Gene osnpf8.1 in Improving Rice Tiller Number

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 OsNPF8.1 Construction of Gene Overexpression Plants

[0021] Extract the RNA of rice Zhonghua 11 and reverse transcribe it into cDNA, using the primer pair:

[0022] F1: 5'- GGTACC ATGGGAGAGGTTGCAGAAGACATC-3'( kpn I),

[0023] R1: 5'- TCTAGA GTTTGCTCCGGCGTGCTCGGCCTT-3'( Xba I);

[0024] Amplified by PCR OsNPF8.1 cDNA of the gene, then passed kpn I. Xba After I digestion, it was connected into the pCAMBIA-1306 vector (the pCAMBIA-1306 vector was purchased from Cambia Company), and the OsNPF8.1 gene overexpression vector OsNPF8.1 -p1306. Agrobacterium EHA105Mediated genetic transformation method, the overexpression vector was introduced into the normal rice variety Zhonghua 11.

[0025] 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 pl...

Embodiment 2

[0032] Example 2 OsNPF8.1 Construction of Gene Interference Plants

[0033] Extract the RNA of rice Zhonghua 11 and reverse transcribe it into cDNA, using the primer pair:

[0034] F4: 5'- GGTACC GGGGTGGTGTTCTTGGCGCTGTAC-3'( Kpn I),

[0035] R4: 5'- GGATCC ACGAGCACCTGCGCGATCCT-3'( Bam H I);

[0036] F5: 5'- ACTAGT GGGGTGGTGTTCTTGGCGCTGTAC-3'( Speech I),

[0037] R5: 5'- GAGCTC ACGAGCACCTGCGCGATCCT-3'( Sac I);

[0038] PCR amplified OsNPF8.1 After the cDNA fragment of the gene was digested with the corresponding restriction endonuclease, it was connected into the pTCK303 vector to construct the OsNPF8.1 gene interference expression vector OsNPF8.1 -pTCK303. Agrobacterium EHA105 Mediated genetic transformation method, introducing the interference expression vector into the normal japonica rice variety Zhonghua 11.

[0039] Transplant all the obtained transgenic seedlings into baskets with soil, water and fertilize them regularly, and plant them in the ...

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Abstract

The invention discloses application of a nitrogen nutrition transport gene OsNPF8.1 in improvement of rice tiller number and belongs to the field of plant genetic engineering. An OsNPF8.1 gene encoded protein has an amino acid sequence shown as SEQ ID NO.1, and a cDNA sequence of the protein is shown as SEQ ID NO.2. By establishing rice OsNPF8.1 gene overexpression plants and OsNPF8.1 gene interference plants, the tiller number of normal rice and the spike number of each rice plant can be increased by improving the OsNPF8.1 gene expression. Therefore, the gene OsNPF8.1 can be used in rice breeding for improving the rice yield. The gene OsNPF8.1 has important application values in the aspect of explaining influences of nitrogen on plant growth and development and the aspect of improving the rice plant type.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, in particular to nitrogen nutrition transport genes OsNPF8.1 Application in increasing rice tiller number. Background technique [0002] China's rice planting area accounts for 20% of the world's total crop planting area, but nitrogen fertilizer application accounts for 37% of the world's total application amount; in 1995, China's nitrogen fertilizer production and use amount reached the first in the world, but nitrogen fertilizer use efficiency is low Low, the amount of nitrogen fertilizer application has increased by 20 times compared with 50 years ago. According to this trend, it is expected to triple again by 2050. Excessive application of nitrogen fertilizer will lead to ecological pollution problems such as eutrophication of water body (Xu Guohua, Fan Xiaorong. Rice nitrate transporter gene OsNRT1.1a and OsNRT1.1b functional research. Nanjing Agricultural University, 2011: 4-6...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8261
Inventor 方中明聂海鹏汪杰吕凯黄玮婷
Owner WUHAN INST OF BIOENG
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