Cucumber (Cucumis sativus L.) fruit protoplast membranes were probed with diphenylhexatriene (DPH) and pyrene, and also with two different plant growth regulators, triacontanol (TRIA) and abscisic acid (ABA). Fluorescence anisotropies of DPH and pyrene were measured after incorporating them into the membranes. The fluorescence lifetime of membrane-bound pyrene was also measured by using neodymium-doped yttrium aluminium garnet (Nd:YAG) laser of 35 ps pulses. The microviscosities of the membranes were calculated using the values of fluorescence anisotropy and lifetime. In the presence of TRIA and ABA, there was a sharp decrease in the fluorescence lifetime of pyrene. Similarly, there was also a decrease in the microviscosities of the membranes and increase in the rate of rotation of membrane-bound fluorophore, induced by the plant growth regulators. Furthermore, TRIA or TRIA + ABA could reduce the fluorescence anisotropy of both the fluorophores whereas, ABA decreased the anisotropy of only pyrene. This property of ABA may be due to its confinement to a specific spacial facet in the membrane. Fatty acid analysis indicated that membrane microviscosity fluctuations were not due to altered fatty acid composition alone as it is known that change in lipid-protein interaction would also alter the physical status of the membrane.