Integrated electrochemical Chip-on-Plant functional sensor for monitoring gene expression under stress

Abstract

The ability to interact with plants, both to sense and to actuate, would open new opportunities for precision agriculture. These interactions can be achieved by using the plant as part of the sensing system. The present work demonstrates real-time monitoring of β-glucuronidase (GUS) expression in transgenic tobacco plants using its activity as a biomarker for functional sensing. As "proof of concept", we demonstrated GUS enzyme biosensing under constitutive expression in Msk8 tomato cells and transgenic tobacco plants and in heat shock inducible BY2 tobacco cells and tobacco plants. The sensing was done using a three-electrode microchip in Msk8 or BY2 cell culture or in tobacco plant leaves. The electrode microchip was used to transduce the expression of the GUS enzyme by chronoamperometry to a measurable electrical current signal. For the constitutive expression of GUS in Msk8 cells, the system sensitivity was 0.076 mA/mM-cm² and the limit of detection was 0.1 mM. For the heat shock inducible BY2 cells the GUS enzyme activity was detected 12-26 h after the heat shock was applied (40 °C for 2 h) using two different substrates: p-nitrophenyl-β-glucuronide (with sensitivity of 0.051 mA/mM-cm²) and phenolphthalein-β-glucuronide (with sensitivity of 0.029 mA/mM-cm²).

Keywords: Electrochemical biosensing; Integrated chip; Plant cell sensor; Precision agriculture.