Abstract
In order to develop a high-efficiency and reproducible transformation protocol for Stylosanthes guianensis we assessed the biological and physical parameters affecting plant electroporation protoplasts. Energy input, as combinations of electric field strengths discharged by different capacitors, electroporation buffer and DNA form were evaluated. Transformation efficiency was assayed in vivo as transient reporter gene expression, using the GFP-coding gene mgfp5 driven by a CaMV 35S constitutive promoter. Energy input and electric field strength had a critical influence on transgene expression with higher transformation levels being achieved with 250 V.cm-1 discharged by 900 and 1000 muF capacitors. Linear plasmid DNA, the absence of chloride and the presence of calcium ions also increased transient gene expression, albeit not significantly.
Highlights
The genus Stylosanthes (Fabaceae) comprises approximately 50 predominantly herbaceous species and subspecies native to tropical and subtropical regions of Asia, Africa and the Americas, principally South America
We used the transient expression of a gene coding for a green fluorescent protein (GFP) (Chalfie et al, 1994) for the in vivo evaluation of transformation efficiency
Maximum transient gene expression has been reported under electric field strengths causing more than 50% reduction in protoplast viability (Fromm et al, 1985; Hauptmann et al, 1987; Oard et al, 1989)
Summary
The genus Stylosanthes (Fabaceae) comprises approximately 50 predominantly herbaceous species and subspecies native to tropical and subtropical regions of Asia, Africa and the Americas, principally South America. Protoplast electroporation allows the introduction of foreign DNA into a great variety of cells, and these cells can be regenerated into transformed plants depending on the effectiveness of the shoot regeneration protocol. Energy input, DNA form and electroporation buffer were assessed in order to optimize transformation rates.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
