Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Genetic transformation method of eucalyptus with selection marker visible to naked eyes under visible light

A genetic transformation method and screening marker technology, applied in biochemical equipment and methods, horticultural methods, botanical equipment and methods, etc., can solve problems such as relying on indirect detection

Inactive Publication Date: 2021-11-05
GUANGDONG UNIV OF PETROCHEMICAL TECH
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is characterized in that it solves the problem that quantitative detection needs to rely on indirect detection, and invents a method for genetic transformation of eucalyptus with screening markers visible to the naked eye under visible light

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 eucalyptus with selection marker visible to naked eyes under visible light
  • Genetic transformation method of eucalyptus with selection marker visible to naked eyes under visible light

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0022] The present invention is further described below.

[0023] (1) Obtain the aseptic vaccine of Eucalyptus urophylla

[0024] (2) Remove the terminal buds and roots of the aseptic seedlings of Eucalyptus urophylla, cut the hypocotyls into 0.2-0.5 cm long sections without bud points, and inoculate them in callus induction medium (MS+PBU 1mg / L+IAA 0.05mg / L+Vc 100mg / L+acetosyringone 0.1mg / L+sucrose 30g / L+agar 6g / L, pH 5.8-6.0). Culture in dark for one week at 24±2°C.

[0025] (3) After the callus is cultured, put the callus in the dipping medium (MS+sucrose 30g / L), ultrasonically shake for 30 seconds, add the Agrobacterium with the red protein reporter gene RUBY, and keep it in the dark at 28°C and 200r shaking Shake in bed for 3h.

[0026] (4) Transfer the soaked callus to callus induction medium (MS+PBU 1mg / L+IAA 0.05mg / L+Vc 100mg / L+acetosyringone 0.1mg / L+sucrose 30g / L+agar 6g / L , pH 5.8-6.0) at 24±2°C for 4-5 days in the dark.

[0027] (5) Transfer the above callus to...

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 belongs to a genetic transformation method of eucalyptus, and particularly relates to a genetic transformation method of eucalyptus for screening positive transformants by naked eyes under visible light. The method comprises the following steps: by taking a stem segment of an aseptic seedling of Eucalyptus urophylla Grandis as an explant to induce a callus, transforming the callus of Eucalyptus urophylla Grandis by using engineering bacteria containing a reporter gene RUBY through an agrobacterium impregnation method, and carrying out adventitious bud induction, bud proliferation, adventitious bud elongation, regenerated seedling rooting and transplanting, so as to obtain a regenerated plant with the red protein reporter gene RUBY visible to naked eyes under natural light. The method is characterized in that the red reporter gene RUBY visible to naked eyes is used as a selection marker, other instruments and sample treatment are not needed, and the method is a non-destructive and convenient method for identifying the positive transformants. The regenerated plant of the red protein reporter gene RUBY visible to naked eyes, which is obtained by adopting the method disclosed by the invention, is stable in result and good in conversion rate, can be directly applied to naked eye observation of whether a target gene is successfully introduced or not, and also lays a foundation for eucalyptus gene breeding.

Description

technical field [0001] The invention relates to a method for genetic transformation of eucalyptus with screening markers visible to the naked eye under visible light, and belongs to the technical field of genetic engineering. Background technique [0002] Agrobacterium is a Gram-negative bacterium ubiquitous in soil, which can chemotactically infect the injured parts of most dicotyledonous plants under natural conditions and induce crown galls or hairy roots. The cells of Agrobacterium tumefaciens and Agrobacterium rhizogenes contain Ti plasmid and Ri plasmid respectively, and there is a segment of T-DNA on it. After Agrobacterium enters the cell by infecting the plant wound, the T-DNA can be inserted into the plant genome. Therefore, Agrobacterium is a natural plant genetic transformation system. People insert the target gene into the modified T-DNA region, and realize the transfer and integration of the exogenous gene to the plant cells by means of Agrobacterium infection...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00A01H6/00A01H4/00
CPCC12N15/8212A01H4/002A01H4/008
Inventor 欧阳乐军李莉梅刘亚梅王泽琛梁楚炎潘璟茵余珠苏敏
Owner GUANGDONG UNIV OF PETROCHEMICAL TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com