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Application of amino acid transport gene OsAAP4 to rice breeding

An amino acid and gene technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve problems such as growth and development without any research, achieve the effect of enhancing tillering ability and improving plant yield

Active Publication Date: 2017-02-22
WUHAN INST OF BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few reports on the research on rice AAP family members, the rice amino acid transport family OsAAP4 There is no research on the effect of genes on the growth and development of rice

Method used

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  • Application of amino acid transport gene OsAAP4 to rice breeding
  • Application of amino acid transport gene OsAAP4 to rice breeding
  • Application of amino acid transport gene OsAAP4 to rice breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 OsAAP4 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 ATGGACAGGAGAGCAGTAGTGTAT-3'( Kpn I),

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

[0024] Amplified by PCR OsAAP4 After cDNA of the gene, 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 OsAAP4 gene overexpression vector OsAAP4 -p1306. Using the genetic transformation method mediated by Agrobacterium EHA105, 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 ...

Embodiment 2

[0032] Example 2 OsAAP4 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'-A GGATCC GAGACAAGGGACGGTGTGGA-3'( Bam H I),

[0035] R4: 5'-AA GGTACC ACCCACACAACATCACGTTCTTCG-3'( Kpn I);

[0036] F5: 5'-A GAGCTC GAGACAAGGGACGGTGTGGA-3'( Sac I),

[0037] R5: 5'-AA ACTAGT ACCCACACAACATCACGTTCTTCG-3'( Speech I);

[0038] PCR amplified OsAAP4 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 OsAAP4 gene interference expression vector OsAAP4 -pTCK303. Using the genetic transformation method mediated by Agrobacterium EHA105, the interference expression vector was introduced into the normal japonica rice variety Zhonghua 11.

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

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PUM

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Abstract

The invention discloses application of the amino acid transport gene OsAAP4 to rice breeding and belongs to the field of plant genetic engineering. The amino acid sequence of OsAAP4 gene encoded protein is as shown in SEQ ID NO.1. The cDAN sequence is as shown in SEQ ID NO.2. By establishing a rice OsAAP4 gene overexpression plant and an OsAAP4 gene interference plant, it is found that the normal rice tiller number and the panicle number of each plant can be increased through improvement of OsAAP4 gene expression, and accordingly the OsAAP4 gene can be used in rice breeding to increase the rice yield. The OsAAP4 gene has important application value in the aspect of stating influences of amino acid transport on plant growth and development process.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, in particular to amino acid transport genes OsAAP4 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. Annu Rev Plant Biol and Plant Mol Biol, 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 (SonodaY, Ikeda A, Saiki S, et al. Feedback regulation of the ammonium transportergene family AMT1 by glutamine in rice. Plant Cell Physiol, 200...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415A01H5/00
CPCC12N9/14C12N15/8205C12N15/8261C12Y306/03
Inventor 方中明吕凯黄玮婷吴博文
Owner WUHAN INST OF BIOENG
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