Gradients in stomatal conductance, xylem sap ABA and bulk leaf ABA along canes of Vitis vinifera cv. Shiraz: molecular and physiological studies investigating their source

Gradients were observed in xylem sap ABA and in stomatal conductance along canes of Vitis vinifera L. cv. Shiraz. To investigate the source of the ABA responsible for these gradients a series of girdling and decapitation experiments were carried out. Leaf stomatal conductance and bulk ABA of leaves and apices were measured in control plants and in response to apex removal or girdling. Gradients in leaf ABA were observed over the first eight expanded leaves of field-grown Shiraz, with higher concentrations of ABA observed towards the apex. Gradients in stomatal conductance that correlated negatively with the concentration of ABA in the leaf ([ABA]leaf) were also observed over the first eight leaves. No significant effect of decapitation was observed on either leaf ABA or stomatal conductance except for the leaf immediately below the apex where a transient increase in [ABA]leaf was observed after 24 h with no corresponding decrease in conductance. Girdling resulted in an increase in [ABA]leaf in leaves distal to the girdle without the corresponding effect on conductance. These effects were further studied at the level of gene activity. To facilitate this, gene sequences encoding two key enzymes involved in the biosynthetic pathway of ABA in grape, zeaxanthin epoxidase (Zep) and 9-cis-epoxycarotenoid dioxygenase (NCED), were isolated and characterised. The cDNA sequences were used as probes to measure the abundances of their respective mRNAs in the leaf and apical material. Levels of expression of one of the two genes encoding NCED, VvNCED1, reflected the gradients in [ABA]leaf in control vines, however treatment-induced changes in ABA were not always associated with corresponding changes in VvNCED1 expression. The abundances of both the VvNCED2 mRNA and Zep mRNA increased with increasing leaf age and did not appear to be associated with either the [ABA]leaf or the expression of VvNCED1. Our results indicate that observed gradients in g s are correlated with [ABA] gradients in mature leaves and xylem sap and that these [ABA] gradients are not derived directly from the apical tissues but, at least partially, from local synthesis.