Electroporation-Like DBD Jet as Rapid FC Delivery Method for Plant Transformation

Plant transformation has become a crucial step in developing both transgenic and non-transgenic crops. It is also a crucial practice wherein foreign constructs (FC) (DNA, RNA, Protein) can be introduced into viable plant cells. A non-thermal atmospheric-pressure dielectric barrier discharge (DBD) jet, driven by a sinusoidal waveform power supply and generating nanoseconds pulse plasma, has been researched in present work as a novel plant transformation delivery system to transfer FC into viable plant cells (here are mature embryo and whole seeds). This is based on a common microbiology technique called electroporation, wherein a high electric field near the cell induces pores to open in the cellular membrane. Here we tested the ability to induce pores in the significantly thicker plant cell wall as well, which enables viable plant transformation. The successful trial on this thicker plant cell is potentially attributed to the generation of a higher electric field compared with traditional electroporation methods [1]–[3] and much higher pulse duration. A serial of plasma characterization has been performed, including power measurement, short-time behavior investigation (time-resolved image), Lissajous analysis, and vibrational & rotational temperature measurements. After exposure to plasma DBD jet, microscope images were taken, indicating 50% and 20% transformation efficiency for mature embryos and whole seeds, respectively. This technique shows promise to significantly shorten the plant transformation time.<br/>Reference:<br/>[1] I. I. Ozyigit, “Gene transfer to plants by electroporation: methods and applications,” Molecular Biology Reports, vol. 47, no. 4. pp. 3195–3210, 2020.<br/>[2] T. E. V. Aune and F. L. Aachmann, “Methodoogies to increase the transformation efficiencies and the range of bacteria that ca nbe transformed.pdf,” Appl. Microbiol ogy Biotechnol., vol. 85, pp. 1301–1313, 2010.<br/>[3] N. K. Sharma, S. Misra, Varun, and U. N. Pal, “Experimental and simulation analysis of dielectric barrier discharge based pulsed cold atmospheric pressure plasma jet,” Phys. Plasmas, vol. 27, no. 11, 2020.