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Application of amino acid transport gene OsANT1 in paddy rice breeding

An amino acid, rice technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problem of no research on rice growth and development, and achieve the effect of increasing plant height and fresh weight.

Active Publication Date: 2017-05-31
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 AAT family members. ANT The protein expressed by the gene mainly transports neutral amino acids, and the current rice amino acid transporter family OsANT1 There is no research on the effect of gene protein on the growth and development of rice

Method used

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  • Application of amino acid transport gene OsANT1 in paddy rice breeding
  • Application of amino acid transport gene OsANT1 in paddy rice breeding
  • Application of amino acid transport gene OsANT1 in paddy rice breeding

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Example 1 OsANT1 Construction of Gene Overexpression Plants

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

[0023] F1: 5'- AGATCT ATGGAGGGGGCGGCGCCGCTTCT-3'( Bgl II),

[0024] R1: 5'- CTTAAG CTGCAGTTGAATCCTGCATTGTGG-3'( Afl II);

[0025] Amplified by PCR OsANT1 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 OsANT1 gene overexpression vector OsANT1 -p1301. Using the genetic transformation method mediated by Agrobacterium EHA105, the overexpression vector was introduced into the normal rice variety Zhonghua 11.

[0026] Transplant all the transgenic seedlings obtained into baskets with soil, water regularly, and fertilize. When the seedlings grow about 10 cm in height, soak 50 rice transgenic seedlings in 500 mL of hygromycete with a concentration of 50 mg / L p...

Embodiment 2

[0031] Example 2 OsANT1 Construction of Gene Interference Plants

[0032] Extract the RNA of rice Zhonghua 11 and reverse transcribe it into cDNA, using the primer pair:

[0033] F3: 5'- GGTACC GAGTGACTCCGGAAGCCAAGC-3'( Kpn I),

[0034] R3: 5'- GGATCC CAGTAGAGCGTGGCGCATCC-3'( Bam H I);

[0035] F4: 5'- ACTAGT GAGTGACTCCGGAAGCCAAGC-3'( Speech I),

[0036] R4: 5'- GAGCTC CAGTAGAGCGTGGCGCATCC-3'( Sac I);

[0037] PCR amplified OsANT1 The cDNA fragment of the gene was digested by the above-mentioned corresponding restriction endonucleases and then connected into the pTCK303 vector to construct the OsANT1 gene interference expression vector OsANT1 - pTCK303. Using the genetic transformation method mediated by Agrobacterium EHA105, the interference expression vector was introduced into the normal japonica rice variety Zhonghua 11.

[0038] Transplant all the transgenic seedlings obtained into baskets with soil, water regularly, and fertilize. When the seedlin...

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Abstract

The invention discloses application of amino acid transport gene OsANT1 in paddy rice breeding, and belongs to the field of plant gene engineering. The amino acid sequence of the OsANT1 gene encoded protein is shown as SEQ ID NO. 1, and the sequence of cDNA is shown as SEQ ID NO. 2. The increase of OsANT1 gene expression is discovered by constructing an overexpression plant of paddy rice OsANT1 gene, and the root length, the root quantity, the plant height and the fresh weight of normal paddy rice can be increased, so that the biomass is improved. The decrease of OsANT1 gene expression is discovered by constructing interference plants, and the root length, the root quantity and the fresh weight of normal paddy rice can be decreased. Therefore, the OsANT1 gene can be used for promoting the growth of paddy rice and increasing the biomass of paddy rice in paddy rice breeding. The OsANT1 gene has significant application values in the aspects of explaining plant growth and development process influenced by amino acid transportation.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, in particular to amino acid transport genes OsANT1 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): C12N15/82C07K14/415C12N15/29A01H5/00
CPCC07K14/415C12N15/8205C12N15/8261
Inventor 方中明吕凯魏倩吴洁刘静钱焌杰
Owner WUHAN INST OF BIOENG
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