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Application of ZmPHR1 protein in regulating and controlling phosphorus content of corn

A technology of phosphorus content and protein, applied in the biological field, can solve the problems of lack of grain, lack of row, poor pollination and so on

Active Publication Date: 2022-05-24
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Phosphorus deficiency during the pollination period will result in poor pollination and curled ears, resulting in missing rows, grains or bald tips, and quality decline

Method used

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  • Application of ZmPHR1 protein in regulating and controlling phosphorus content of corn
  • Application of ZmPHR1 protein in regulating and controlling phosphorus content of corn
  • Application of ZmPHR1 protein in regulating and controlling phosphorus content of corn

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1, the discovery of the ZmPHR1 protein and its coding gene (i.e., ZmPHR1 gene).

[0072] 1. The total RNA of maize inbred line B73 was extracted by TRizol method. The integrity of the total RNA of maize inbred line B73 was checked by formaldehyde-denaturing RNA agarose gel electrophoresis.

[0073] 2. Take the total RNA of the corn inbred line B73, according to SUPERSCRIPT II Instructions for use (Thermo Fisher Scientific products) synthetic corn inbred B73 single-stranded cDNA.

[0074] 3. Take the single-stranded cDNA of maize inbred B73, dilute it 10 times with water and use it as a template, and use Primer 1:5'-tataagcttATGAGGAACTTTAATCTGATGCAGT-3' and Primer 2:5'-gctctagaTTAACTTTTGCGCGC-3' primer pairs for PCR amplification, and recover pcR amplification products of about 1370bp.

[0075] The reaction system is 50 μL, consisting of 10 μL 5×Phusion HF Buffer, 4 μL 2.5 mM dNTP mix, 2.5 μL Primemer 1 aqueous solution (concentration 10 μM), 2.5 μL Primer 2 aqueous ...

Embodiment 2

[0080] Example 2, T 3 Acquisition and phenotypic identification of ZmPHR1 gene corn by generation homozygous

[0081] First, the construction of recombinant plasmid UBI: ZmPHR1

[0082] 1. Using the recombinant plasmid pMD18-ZmPHR1 as a template, the primer pair composed of primer 1:5'-ATGAGGAACTTTAATCTGATGCAGT-3' and primer 2:5'-TTAACTATCTTGCTTGCGC-3' was used to pcR amplify the PCR amplification product.

[0083] 2. Take the PCR amplification product obtained in step 1, use Taq enzyme to supplement A at the flat end, and obtain the PCR amplification product with A sticky end.

[0084] 3. Linearized vector pCXUN was obtained by using the restriction enzyme XcmI (NEB company) to digest the vector pCXUN (GenBank: FJ905215.1). The linearized carrier pCXUN has a T-stick end.

[0085] 4. By TA cloning method, the PCR amplification product with A sticky end and the linearized carrier pCXUN are connected to obtain the recombinant plasmid UBI:ZmPHR1.

[0086]Sequence the recombinant plas...

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Abstract

The invention discloses an application of a ZmPHR1 protein in regulating and controlling the phosphorus content of corn. The ZmPHR1 gene is introduced into a corn selfing line B73, and then T3-generation homozygous ZmPHR1 gene transgenic corn is obtained through selfing. And detecting the content of inorganic phosphorus in the crown of the T3-generation homozygous ZmPHR1 transgenic corn, and counting the hundred-grain weight. Under the condition of sufficient phosphorus or low phosphorus, compared with a corn inbred line B73, the crown inorganic phosphorus content and hundred-grain weight of T3-generation homozygous ZmPHR1 transgenic corn are remarkably increased. Therefore, the ZmPHR1 protein can regulate and control the phosphorus content and hundred-grain weight of the corn, and further regulate and control the yield. The method has an important application value.

Description

Technical field [0001] The present invention belongs to the field of biotechnology, specifically relates to the application of ZmPHR1 protein in regulating the phosphorus content of corn. Background [0002] Phosphorus is one of the necessary nutrients for plant growth and development, participates in a variety of life activities, and is an important component of nucleic acids, nucleoproteins, phospholipids and other functional substances. Phosphorus is involved in the energy metabolism of plant cells, the synthesis and catabolism of various organic matter, and also participates in life activities such as plant cell signal transduction. [0003] The phosphorus concentration in plant cells is generally maintained at the mM level, while the concentration of available phosphorus in the soil solution is extremely low, generally less than 10 μM, plants / crops often face low phosphorus stress, and soil phosphorus deficiency has become an important limiting factor for agricultural produc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84C07K14/415C12N15/29A01H5/00A01H6/46
CPCC12N15/8261C12N15/8243C07K14/415A01H6/4684A01H5/00Y02A40/146
Inventor 陈益芳武维华王海峰李小梅
Owner CHINA AGRI UNIV