Genetic transformation method of white poplars

A genetic transformation method, the technology of Populus alba, which is applied in the field of genetic transformation of Poplar alba, can solve the problems of shortening the transformation time and the inapplicability of genetic transformation of Poplar alba, and achieve the effect of shortening the transformation time and improving the transformation efficiency

Active Publication Date: 2019-12-13
CHINESE ACAD OF FORESTRY
View PDF1 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In view of this, the purpose of the present invention is to provide a kind of poplar genetic transformation method, solve the inapplicability problem of existing poplar g

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Genetic transformation method of white poplars
  • Genetic transformation method of white poplars
  • Genetic transformation method of white poplars

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0037] In the present invention, the explants are soaked in the infecting bacteria solution, and the soaking time is preferably 10-15 minutes. The preparation method of the infection bacterium liquid described in the present invention preferably comprises the following steps: A. Agrobacterium carrying the recombinant plasmid is streaked on the YEB plate medium to obtain a single colony; the YEB plate medium contains 50 mg / L of kanamycin and 50mg / L of rifampicin;

[0038]B. Inoculate the single colony according to the ratio of 1 single colony: 5mL YEB liquid medium, and culture with shaking until OD 600 0.7~0.9, to get the expanded strain; the YEB liquid culture medium contains the kanamycin of 50mg / L and the rifampicin of 50mg / L;

[0039] C. Inoculate the expanded strains in YEB anti-resistant liquid medium, and culture with shaking until OD 600 0.7 to 0.9 to obtain an activated bacterial liquid; the volume ratio of the expanded strain to the YEB-free liquid medium is 1 μL:...

Embodiment 1

[0061] Transferring the GUS gene into Populus alba:

[0062] 1. Connect the GUS gene (GenBank: MG687280.1) to the ΔpCAMBIA1302 vector. The ΔpCAMBIA1302 vector is a new binary expression vector transformed from the traditional pCAMBIA1302. It mainly uses the CaMV35S promoter (CaMV35S-P) (835bp) and multiple The cloning site (42bp) replaced the constitutive CaMV35S promoter (CaMV35S-P) (538bp) and the reporter gene GFP on the original T-DNA segment of the pCAMBIA1302 vector. Transfer the recombinant gene into DH10B Escherichia coli competent. After the sequencing was correct, the plasmid was extracted and transformed into EHA105 Agrobacterium competent by electric shock transformation.

[0063] 2. Streak the Agrobacterium (EHA105) carrying the recombinant plasmid on a YEB (50mg / L Kan+50mg / L Rif) plate, culture at 28°C, and a single colony appears after 2 days.

[0064] 3. Pick a single colony for colony PCR, and inoculate it in 1mL YEB (50mg / L Kan+50mg / LRif) liquid medium, and...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a genetic transformation method of white poplars, and relates to the technical field of genetic transformation. In the present invention, target genes are transferred into the body of the white poplars through an agrobacterium infection method to obtain positive plants in a short time. In an embodiment of the present invention, beta-glucuronidase genes (GUS) are used as reporter genes to carry out genetic transformation of the white poplars, so that a large number of transgenic white poplars can be obtained in a short time. The genetic transformation method of the whitepoplars solves the problem that the existing genetic transformation method for poplars is not applicable in genetic transformation of the white poplars. The genetic transformation method of the whitepoplars can greatly improve transformation efficiency of the white poplars and effectively shorten transformation time.

Description

technical field [0001] The invention belongs to the technical field of genetic transformation, and in particular relates to a genetic transformation method of Populus alba. Background technique [0002] Populus alba (PopμLμs abla L.) is widely distributed in the western Mediterranean basin, Central Europe, Eastern Europe and the valleys of Central Asia. In my country, it is mainly distributed in the south of Liaodong, Shaanxi, Ningxia, Gansu, Shanxi, Qinghai, Xinjiang and Tibet. It is the main afforestation tree species in the Qinghai-Tibet Plateau and the Northwest Loess Plateau. It is often used for farmland protection, road greening, barren sand and wasteland management, building materials and firewood, furniture and carving patterns, etc. In the 1980s, my country began the research on tissue culture and rapid propagation of Populus alba, and made important progress. However, the research on the transformation system of Populus alba is not very mature at present, especia...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N15/82C12N15/65A01H4/00A01H5/00A01H6/00
CPCA01H4/00C12N15/65C12N15/8205
Inventor 曾庆银刘妍婧姜鹏飞王晓霞王一鸣
Owner CHINESE ACAD OF FORESTRY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap