Application of amino acid transporter gene OsAAP3 in rice seed selection

An amino acid, rice technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problem of no research on the growth and development of rice, and achieve the effect of enhancing the tillering ability, increasing the yield of the plant, and increasing the number of tillers.

Active Publication Date: 2017-03-22
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 OsAAP3 The protein of the gene can transport various amino acids such as lysine and arginine (Taylor M R, ReindersA, Ward J M. Transport function of rice amino acid permeases (AAPs)[J]. Plant and Cell Physiology, 2015, 56(7) : 1355-1363.), but there is no research on the growth and development of rice

Method used

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  • Application of amino acid transporter gene OsAAP3 in rice seed selection
  • Application of amino acid transporter gene OsAAP3 in rice seed selection
  • Application of amino acid transporter gene OsAAP3 in rice seed selection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 OsAAP3 Construction of Gene Interference Plants

[0019] Extract the RNA of rice Zhonghua 11 and reverse transcribe it into cDNA, using the primer pair:

[0020] F1: 5'- GGTACC AAGGACGTGGAGATGGCG-3'( Kpn I),

[0021] R1: 5'- GGATCC ATCGCCAGTGCGGTAGCAGTC-3'( Bam H I);

[0022] F2: 5'- ACTAGT AAGGACGTGGAGATGGCG-3'( Speech I),

[0023] R2: 5'- GAGCTC ATCGCCAGTGCGGTAGCAGTC-3'( Sac I);

[0024] PCR amplified OsAAP3 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 OsAAP3 gene interference expression vector OsAAP3 -pTCK303. Using the genetic transformation method mediated by Agrobacterium EHA105, the interference expression vector was introduced into the normal japonica rice variety Zhonghua 11.

[0025] Transplant all the obtained transgenic seedlings into baskets with soil, water and fertilize them regularly, and plant them i...

Embodiment 2

[0032] Example 2 OsAAP3 Construction of Gene Overexpression Plants

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

[0034] F5: 5'- AGATCT ATGGCGAAGGACGTGGAGATGGCG-3'( Bgl II),

[0035] R5: 5'- CTTAAG TCACGACTTGGTCTTGAAGGGGACGTA-3'( Afl II);

[0036] Amplified by PCR OsAAP3 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 OsAAP3 gene overexpression vector OsAAP3 -p1301. Using the genetic transformation method mediated by Agrobacterium EHA105, the overexpression vector was introduced into the normal rice variety Zhonghua 11.

[0037] 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 dete...

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Abstract

The invention discloses application of an amino acid transporter gene OsAAP3 in rice seed selection, and belongs to the technical field of genetic engineering. An amino acid sequence of encoded protein of the gene OsAAP3 is specified in SEQ ID NO.1, and a Cdna sequence is specified in SEQ ID NO.2. According to the application of the amino acid transporter gene OsAAP3 in rice seed selection, by constructing a rice plant which is interfered by the gene OsAAP3, and constructing a rice plant which is over-expressed by the gene OsAAP3, it is discovered that reduction of genetic expression of the gene OsAAP3 can increase tiller number of the rice and spike number in each rice plant, and therefore the gene OsAAP3 can be used in rice seed selection for increasing the rice yield. The gene OsAAP3 has important application value in expounding influence of amino acid transportation on growth and development process of the plant.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, in particular to amino acid transport genes OsAAP3 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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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29A01H5/00
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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