ABA Transport Assay in Plant Single-Cell System.

Compare with long history of the abscisic acid (ABA) study, our knowledge of ABA transporters is quite recent. This is due to at least one reason: ABA is a weak acid, and thus, it exists in either protonated form or in anionic form depending on the surrounding pH relative to its pKa value. Because the protonated form of ABA can permeate the cell membrane, it would foreclose a specific uptake transporter of ABA. Notwithstanding this theoretical base, ABA transporters belonging to different protein families have been reported a decade ago, steadily. A critical point of the identification of novel ABA transporters is to prove their transport activity. To do this, heterologous expression system is considered first as a facility of transport activity analysis. However, it is difficult to overexpress membrane proteins in their functional state in heterologous system. They have the tendency to aggregate and produce inclusion bodies caused by mistargeting. Thus, in this chapter, I describe the method of ABA transport assay based on single-cell system originated from plant tissues.