Detached and intact leaves (first fully expanded leaf from the top) of tobacco (Nicotiana tabaccum L.) plantlets hardened in vitro with 2.0% polyethylene glycol (PEG) showed increased diffusive resistance (r) over those of nonhardened plantlets as measured by a steady state porometer. The leaves of the PEG hardened plants maintained a higher resistance throughout the one hour dessication period in approximately 30% relative humidity although both treatments showed an increase in diffusive resistance after 30 minutes. This indicates that the stomates are functioning in the in vitro tobacco plantlets. The higher (r) in the PEG treated plants may be due to more complete closure of stomates, higher cuticle wax content or a combination of both.