Application of nitrate radical transporter gene OsNRT1.8 in rice breeding

A technology of osnrt1.8 and 1.osnrt1.8, applied in the application field of nitrate transport gene OsNRT1.8 in rice breeding, to achieve the effect of enhancing tillering ability and improving plant yield

Active Publication Date: 2017-01-18
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 OsNRT1.8 The gene is a nitrate transport homolog of the rice NPF gene family

Method used

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  • Application of nitrate radical transporter gene OsNRT1.8 in rice breeding
  • Application of nitrate radical transporter gene OsNRT1.8 in rice breeding
  • Application of nitrate radical transporter gene OsNRT1.8 in rice breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 OsNRT1.8 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'- AGATCT GTCATGGACGCCGGAGACGCCAT-3'( Bgl II),

[0023] R1: 5'- CTTAAG GACGTCTCACGAGAGCACGGTCT-3'( Afl II);

[0024] Amplified by PCR OsNRT1.8 After cDNA of the gene, passed Bgl II and Afl After digestion with II, it was connected into the pCAMBIA-1301 vector (the pCAMBIA-1301 vector was purchased from Cambia Company), and the OsNRT1.8 gene overexpression vector OsNRT1.8 -p1301. Agrobacterium EHA105 Mediated 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 t...

Embodiment 2

[0032] Example 2 OsNRT1.8 Construction of gene interference plants and OsNRT1.8 Acquisition of genetic mutants

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

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

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

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

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

[0038] PCR amplified OsNRT1.8 After the cDNA fragment of the gene was digested by the above-mentioned corresponding restriction endonucleases, it was connected into the pTCK303 vector to construct the OsNRT1.8 gene interference expression vector OsNRT1.8 -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 ...

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Abstract

The invention discloses application of a nitrate radical transporter gene OsNRT1.8 in rice breeding and belongs to the field of plant genetic engineering. The protein encoding amino acid sequence of the gene OsNRT1.8 is shown as SEQ ID NO.1, and a cDNA sequence is shown as SEQ ID NO.2. By establishing rice gene OsNRT1.8 overexpression plant and a gene OsNRT1.8 interference plant, it is found that normal rice tiller number and spike number of each plant can be increased by improving gene OsNRT1.8 expression. Therefore, the gene OsNRT1.8 can be used for increasing rice yield in rice breeding. The gene OsNRT1.8 has an important application value on the aspects of elaboration about nitrogen influence on plant growth and development and improvement of rice plant type.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, in particular to nitrate transport genes OsNRT1.8 Application in rice breeding. 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 [Xu Guohua, Fan Xiaorong. Rice Nitrate Transporter Gene OsNRT1.1a and OsNRT1.1b functional research. Nanjing Agricultural University, 2011: 4-6]. More nitrogen nutrients are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415A01H5/00
CPCC07K14/415C12N15/8261
Inventor 方中明黄玮婷吕凯王松太
Owner WUHAN INST OF BIOENG
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