Direct protein introduction into plant cells using a multi-gas plasma jet

Area of Science:

• Plant Biology
• Biotechnology
• Biophysics

Background:

• Protein delivery into plant cells is significantly more challenging than into mammalian cells.
• Previous methods often require cell-penetrating peptides for successful protein introduction in plant tissues like leaves.

Purpose of the Study:

• To investigate the efficacy of atmospheric non-thermal plasmas (CO2 or N2) for direct protein introduction into tobacco leaf cells.
• To assess if protein delivery can be achieved without the use of cell-penetrating peptides.

Main Methods:

• Generation of CO2 and N2 atmospheric non-thermal plasmas using a multi-gas plasma jet.
• Treatment of tobacco leaves with the generated plasmas.
• Introduction of superfolder green fluorescent protein (sGFP)-fused adenylate cyclase as a reporter protein.
• Confocal microscopy for sGFP signal detection within leaf cells.
• Measurement of cyclic adenosine monophosphate (cAMP) levels to confirm enzyme activity.

Main Results:

• Superfolder green fluorescent protein (sGFP) signals were successfully detected within tobacco leaf cells following CO2 or N2 plasma treatment.
• Plasma treatments did not cause substantial damage to the leaf tissues.
• A significant increase in cyclic adenosine monophosphate (cAMP) was observed, confirming the functional introduction of adenylate cyclase.

Conclusions:

• Atmospheric non-thermal CO2 or N2 plasma treatment is a viable method for introducing functional proteins into plant cells.
• This technique offers a promising, peptide-free alternative for protein delivery in plant tissues.
• Plasma-based protein introduction holds potential for advancing plant biotechnology and research applications.