Digitalized pencil trace modified electrodes for real time evaluation of salicylic acid in detached Arabidopsis thaliana leaves during regeneration.

Abstract Plants have excellent abilities to regenerate from detached tissues, in which various phytohormones play critical roles. It has been reported that jasmonate and auxin appeared sequentially during de novo root regeneration (DNRR) after leave detachment. However, the role of salicylic acid (SA) is still unknown in this procedure although it is another important plant hormone. We have demonstrated the potential of electrochemical sensors for real time screening of SA but the stability still needed to be improved. Herein a digital plotter was used to modify the carbon tape modified electrodes with pencil traces in order to improve the reproducibility. The modified electrodes in paper-based analytical devices were applied to monitor SA during DNRR. The drawing routines and the distances between two close traces were optimized. Our results showed that the carbon tape modified electrodes achieved more reproducible responses of SA. Combined with in vivo sampling, the results using our approach demonstrated that amounts of SA in the wild Arabidopsis thaliana leaves during direct DNRR reached highest at around 72 hr after detachment and then decreased, implying that the wave of SA contents might follow that of auxin during direct DNRR. The application of the digital plotter offered a cost-effective and more reproducible method for preparation of disposable working electrodes, which might be extended for other biochemical assays.