Method for creating phenotypic variation transgenic plant by utilizing methyltransferase gene

A phenotypic variation and gene technology, applied in the field of using methyltransferase gene to create phenotypic variation transgenic plants, can solve problems such as lack of plant breeding germplasm resources

Inactive Publication Date: 2020-02-11
INST OF FORESTRY CHINESE ACAD OF FORESTRY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In order to solve the problem of lack of germplasm resources in plant breeding, the present invention provides a method for creating phenotype-vari

Method used

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  • Method for creating phenotypic variation transgenic plant by utilizing methyltransferase gene
  • Method for creating phenotypic variation transgenic plant by utilizing methyltransferase gene
  • Method for creating phenotypic variation transgenic plant by utilizing methyltransferase gene

Examples

Experimental program
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Embodiment 1

[0040] Example 1: Acquisition of AtMET1-transferred 84K poplar and phenotypic variation transgenic plants

[0041] 1. Construction of a chemically inducible plant expression vector for Arabidopsis methylation gene AtMET1

[0042] According to the AtMET1 sequence in GenBank, full-length primers MET1-F (5'-CCGCTCGAGATGGTGGAAAATGGGGCTA-3') and MET1-R (5'-CTAGACTAGTCTAGGGTTGGTGTTGAGGAG-3') with Xho I and Spe I restriction sites were designed to carry AtMET1 The plasmid of the gene was used as a template, and PCR amplification was performed to amplify the full-length AtMET1 gene. After detection by 1% agarose gel electrophoresis, the target fragment was recovered and purified with the Axygen company's agarose gel recovery kit, and connected to the cloning vector pMDTM19 -T Vector (Takara Company, Japan), transformed E.coli DH5α, obtained pMDTM19-T-AtMET1 recombinant plasmid, after PCR and double enzyme digestion, it was confirmed that the insert size was correct, which was 4,605bp....

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Abstract

The invention belongs to the field of plant breeding, relates to the field of plant biotechnology breeding, and particularly relates to a method for creating a phenotypic variation transgenic plant byutilizing a methyltransferase gene. According to the method, an agrobacterium-mediated method is used for introducing a chemical induction promoter and the arabidopsis DNA methyltransferase gene AtMET1 into an 84K poplar genome to obtain an AtMET1 84K poplar plant, a sterile in-vitro leaf AtMET1 of the transgenic plant is induced to express for 3 hours, 12 hours and 24 hours by using estrogen, and then the leaf is induced to regenerate to obtain the phenotypic variant plant. According to the research, the chemical induction promoter is used for artificially promoting exogenous gene expression, phenotypic variation germplasm is created, and plant breeding resources are enriched.

Description

technical field [0001] The invention belongs to the field of plant breeding, in particular to the field of plant biotechnology breeding, and specifically relates to a method for creating phenotypic variation transgenic plants by using methyltransferase genes. Background technique [0002] Poplar belongs to the Salicacae family Populus L. and is a perennial woody tree. Poplar is not only an important tree species for afforestation and industrial timber, but also a model tree species for perennial forest genetic engineering. [0003] Since the development of poplar hybrid breeding in my country in the 1940s, a number of poplar varieties with excellent growth performance have been selected and bred, and have been promoted and planted in major domestic cultivation areas, achieving good social and economic benefits. With the development of the economy, the demand for improved poplar varieties is increasing day by day. However, limited by the lack of breeding resources, the effec...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/54C12N15/66C12N1/21A01H4/00A01H5/00A01H6/00C12R1/01
CPCA01H4/00C12N9/1007C12N15/66C12N15/8261
Inventor 张冰玉苏晓华张伟溪常英英吴晓娟武舒
Owner INST OF FORESTRY CHINESE ACAD OF FORESTRY
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