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A highly efficient protocol for transforming Cuscuta reflexa based on artificially induced infection sites
Lena Anna-Maria Lachner1, Levon Galstyan Galstyan1,2, Kirsten Krause1
1Department of Arctic and Marine Biology UiT The Arctic University of Norway Tromsø Norway.
Plant Direct
|August 14, 2020
Summary
A new Agrobacterium-mediated transformation protocol efficiently transforms Cuscuta reflexa cells, overcoming previous challenges. This method utilizes the plant
Area of Science:
- Plant biotechnology
- Parasitic plant research
- Molecular biology
Background:
- The parasitic plant genus *Cuscuta* presents significant challenges for genetic transformation and in vitro propagation.
- Efficient gene expression techniques are crucial for analyzing *Cuscuta*'s host interaction and infection mechanisms, vital for agroecosystem management.
- Current transformation protocols for *Cuscuta* are scarce and inefficient, hindering functional studies.
Purpose of the Study:
- To explore *Agrobacterium*-mediated transformation of *Cuscuta* plant parts as an efficient alternative to existing methods.
- To develop and validate a novel, highly efficient transformation protocol for *Cuscuta reflexa*.
- To identify specific *Cuscuta* tissues or cells most amenable to transformation.
Main Methods:
- Utilized *Agrobacterium rhizogenes* and *Agrobacterium tumefaciens* with binary transformation vectors containing fluorescent protein genes.
- Applied transformation to various *Cuscuta reflexa* plant parts, focusing on the infection organ.
- Employed fluorescent protein markers to assess transformation efficiency and stability.
Main Results:
- Developed a novel, highly efficient transformation protocol for *Cuscuta reflexa* cells.
- Observed high transformation efficiency by exploiting the infection organ's propensity to uptake and express transgenes.
- Identified the cell layer below the adhesive disk's epidermis as highly susceptible to *Agrobacterium* infection.
- Demonstrated stable transgene expression in transformed explants for several weeks in vitro.
- Showcased frequent cotransformation of susceptible cells when using mixed *Agrobacterium* strains.
Conclusions:
- The novel *Agrobacterium*-mediated transformation protocol offers an efficient alternative for transgene expression in *Cuscuta*.
- The infection organ's specific cellular layers exhibit high competence for foreign DNA uptake, facilitating transformation.
- This technique holds promise for future development of transformed *Cuscuta* cells into callus for further research and application.
