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Application of amino acid transport gene OsAAP1 to promotion of rice growth in low nitrogen

An amino acid and gene technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problem of no research on growth and development, and achieve the effect of increasing rice biomass and plant biomass.

Active Publication Date: 2017-07-07
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 on the study of rice AAT family members, and the rice amino acid transport family OsAAP1 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 OsAAP1 to promotion of rice growth in low nitrogen
  • Application of amino acid transport gene OsAAP1 to promotion of rice growth in low nitrogen
  • Application of amino acid transport gene OsAAP1 to promotion of rice growth in low nitrogen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 OsAAP1 Construction of Gene Overexpression Plants

[0020] Extract the RNA of rice Zhonghua 11 and reverse transcribe it into cDNA, using the primer pair:

[0021] F1: 5'- AGATCT ATGGGGATGGAGAGGCCGCAAGAG-3'( Bgl II),

[0022] R1: 5'- CTTAAG TCATGAGGAGACGCTGAATGG-3'( Afl II);

[0023] Amplified by PCR OsAAP1 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 OsAAP1 gene overexpression vector OsAAP1 -p1301. Using the genetic transformation method mediated by Agrobacterium EHA105, the overexpression vector was introduced into the normal rice variety Zhonghua 11.

[0024] Transplant all the obtained transgenic seedlings into baskets with soil, water and fertilize regularly, and when the seedlings grow about 10 cm in height, soak 50 rice transgenic seedlings in 500 mL of distilled water to prepare hygromycin with a...

Embodiment 2

[0025] Example 2 OsAAP1 Construction of Gene Interference Plants

[0026] Extract the RNA of rice Zhonghua 11 and reverse transcribe it into cDNA, using the primer pair:

[0027] F2: 5'- GGTACC TGGGGATGGAGAGGCCGCAA-3'( Kpn I),

[0028] R2: 5'- GGATCC TTGCGCTTGCCATGGACGGGGT-3'( Bam H I);

[0029] F3: 5'- ACTAGT TGGGGATGGAGAGGCCGCAA-3'( Speech I),

[0030] R3: 5'- GAGCTC TTGCGCTTGCCATGGACGGGGT-3'( Sac I);

[0031] PCR amplified OsAAP1 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 OsAAP1 gene interference expression vector OsAAP1 - pTCK303. Using the genetic transformation method mediated by Agrobacterium EHA105, the interference expression vector was introduced into the normal japonica rice variety Zhonghua 11.

[0032] Transplant all the obtained transgenic seedlings into baskets with soil, water and fertilize regularly, and when the se...

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Abstract

The invention discloses application of amino acid transport gene OsAAP1to promotion of rice growth in low nitrogen, belonging to the field of plant genetic engineering. Amino acid of the OsAAP1 gene coding protein and cDNA sequence thereof is shown as following SEQID NO.1, 2. The rice root length, root number, plant height and tiller number can be increased and biomass and yield can be improved by improving the OsAAP1 gene expression in low nitrogen through establishing the rice OsAAP1 overexpression plant. The rice root length, root number, plant height and tiller number can be reduced by decreasing the OsAAP1gene expression through establishing interfering plants. Thereby, the OsAAP1gene can be applied in improving the biomass and the yield of the rice in low nitrogen. The OsAAP1 gene has important application in the respect of rice tolerance to low or poor soil or farmland, reduction of chemical fertilizer usage amount and the rice breed improvement.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, in particular to amino acid transport genes OsAAP1 Applications to promote rice growth under low nitrogen. 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 ric...

Claims

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Application Information

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IPC IPC(8): C12N15/55C12N9/14A01H5/00
CPCC12N9/14C12N15/8205C12N15/8261C12Y306/03
Inventor 吕凯方中明黄玮婷刘静
Owner WUHAN INST OF BIOENG
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