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Corn C4 phosphoric acid enol pyruvic acid carboxylase gene and uses in producing wheat

A technology of pyruvate carboxylase and phosphoenol, which is applied in the fields of application, genetic engineering, and plant gene improvement, and can solve the problems of large-scale genetic transformation research difficulty and excessively large target fragments

Inactive Publication Date: 2009-07-29
HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the total length of the pepc gene used in this study is close to 9kb, and the target fragment is too large, making it difficult to conduct large-scale genetic transformation research on crops such as wheat and rice

Method used

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  • Corn C4 phosphoric acid enol pyruvic acid carboxylase gene and uses in producing wheat
  • Corn C4 phosphoric acid enol pyruvic acid carboxylase gene and uses in producing wheat
  • Corn C4 phosphoric acid enol pyruvic acid carboxylase gene and uses in producing wheat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Corn C 4 Cloning and bioinformatics analysis of phosphoenolpyruvate carboxylase (PEPC) gene

[0035] 1. Materials and methods

[0036] 1.1 Materials

[0037] 1.1.1 Plant materials and reagents

[0038] The maize inbred line material involved in the present invention is provided by the Food Crops Research Institute of Henan Academy of Agricultural Sciences.

[0039] Experimental RNA extraction kits and agarose gel DNA recovery kits were purchased from Tianwei Times Company, MLV reverse transcription kits were purchased from Promega Company, LA-Taq DNA polymerase, pMD-19T cloning vector, Escherichia coli (Escherichia coli) strain DH5α competent cell preparation kits are all products of TaKaRa Company, the plant expression vector pCOMBIA3301 is preserved in our laboratory, the restriction endonuclease is the product of MBI Company, the ampicillin is the product of Amersco Company, and the rest of the reagents are imported or domestic analysis pure.

[0040]...

Embodiment 2

[0090] Example 2 Corn C 4 Gene transfer of type phosphoenolpyruvate carboxylase into wheat and its functional analysis

[0091] 1. Materials and methods

[0092] 1.1 Materials

[0093] 1.1.1 Plant materials and reagents

[0094] Five wheat varieties (lines): Zhoumai 19, 00H97-54-2-7, Zhengmai 9023, 17-1 and Xinmai 18 were provided by the Molecular Breeding Laboratory of Wheat Research Center, Henan Academy of Agricultural Sciences. Gene gun reagents and consumables were purchased from Bio-Rad, the United States, PCR reagents were purchased from Dalian Bao Biological Company, and the rest of the reagents were imported or domestically produced.

[0095] 1.2 Method

[0096] 1.2.1 Transformation, screening and regeneration of target genes

[0097] The immature embryos of 5 wheat varieties (lines) were used as recipient materials. Take the immature seeds in the middle of the panicle and the same size 12-14 days after flowering, disinfect the surface with 70% alcohol for 1min,...

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Abstract

The invention relates to a corn C4 type phosphoenolpyruvate carboxykinase gene and an application thereof in wheat production. The gene is as indicated in SEQ ID No:1 of a sequence table, a gene clone vector is pMD19-pepc, an eucaryon expression vector is p3301-pepc, host bacteria is E.coli competent cell DH5 alpha, carboxylase is has an amino acid sequence as indicated by SEQ ID No:2. In the invention, the corn C4 type phosphoenolpyruvate carboxykinase is led into the wheat, functional analysis on transgenic plants indicates that net photosynthetic rate of the transgenic plants is greatly improved compared with the net photosynthetic rate of the plants which are not transgenic, enjoying an increase of 1.39 times. The research is so far the research in which the net photosynthetic rate of C3 crop is raised to the largest degree. The result of the functional analysis of the transgenic plants indicates that the related corn C4 type high light efficiency pepc gene can greatly improve the net photosynthetic rate of C3 crops such as wheat and the like and provides important technological and material support for high light efficiency transgenic breeding of crops such as wheat and the like.

Description

technical field [0001] The invention belongs to the field of biological genes, in particular to a C 4 Type phosphoenolpyruvate carboxylase (PEPC) gene and its application in wheat production. Background technique [0002] Wheat, rice and other crops are the main food crops in my country, and their production capacity and supply and demand have always been related to major strategic issues such as my country's national economic development and food security. From a global perspective, the state of world grain production has always been closely related to the political and economic development of each country. Once grain production and reserves decline, international grain prices will rise sharply, and panic will spread. From a domestic point of view, with the growth of my country's population, the rapid development of industrialization and urbanization, the demand for food will continue to increase. At the same time, since the acreage of grain crops in my country cannot reco...

Claims

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Application Information

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IPC IPC(8): C12N15/60C12N9/88C12N15/82
Inventor 许为钢李艳张磊胡琳
Owner HENAN ACAD OF AGRI SCI
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