Water-stress-induced changes in the abscisic acid content of guard cells and other cells of Vicia faba L. leaves as determined by enzyme-amplified immunoassay

Abstract

A highly sensitive, solid-phase, enzyme-amplified immunoassay for the plant growth regulator (+)-abscisic acid (ABA) was developed. The assay sensitivity (0.2-10 fmol) was sufficient for analyzing free ABA in homogeneous tissue samples dissected from Vicia faba L. leaves. Eight hours after detached leaves had been desiccated to 10% loss of fresh weight, the bulk leaf ABA content increased from </=0.2 to 6.2 ng.(mg dry weight)(-1). Epidermal tissue, spongy parenchyma cells, and palisade parenchyma cells from this water-stressed leaf had the following ABA contents, respectively: 4.8, 9.4, and 9.0 ng.(mg dry weight)(-1). Guard cells, which respond to exogenous ABA by losing solutes and volume, were also assayed. When they were dissected from control (fully hydrated) leaves, their ABA content was approximately 0.7 fg.(cell pair)(-1) [[unk]0.2 ng.(mg dry weight)(-1)]. In contrast, the ABA content of guard cells of water-stressed leaves was approximately 17.7 fg.(cell pair)(-1). These results indicate that ABA accumulation in a highly stressed V. faba leaflet is generalized; guard cells contain only 0.15% of bulk leaf ABA. The time course for loss of ABA from guard cells of a floating epidermal peel was studied. There was little loss within 30 min, but after 4 hr, the ABA content was only 17% of the original value. These results indicate that the bulk of guard cell ABA is not readily diffusible (i.e., probably not apoplastic). The results also indicate that common laboratory procedures results in lowered guard cell ABA content.