Mind the bubbles: achieving stable measurements of maximum hydraulic conductivity through woody plant samples

The maximum specific hydraulic conductivity (k(max)) of a plant sample is a measure of the ability of a plants' vascular system to transport water and dissolved nutrients under optimum conditions. Precise measurements of k(max) are needed in comparative studies of hydraulic conductivity, as well as for measuring the formation and repair of xylem embolisms. Unstable measurements of k(max) are a common problem when measuring woody plant samples and it is commonly observed that k(max) declines from initially high values, especially when positive water pressure is used to flush out embolisms. This study was designed to test five hypotheses that could potentially explain declines in k(max) under positive pressure: (i) non-steady-state flow; (ii) swelling of pectin hydrogels in inter-vessel pit membranes; (iii) nucleation and coalescence of bubbles at constrictions in the xylem; (iv) physiological wounding responses; and (v) passive wounding responses, such as clogging of the xylem by debris. Prehydrated woody stems from Laurus nobilis (Lauraceae) and Encelia farinosa (Asteraceae) collected from plants grown in the Fullerton Arboretum in Southern California, were used to test these hypotheses using a xylem embolism meter (XYL'EM). Treatments included simultaneous measurements of stem inflow and outflow, enzyme inhibitors, stem-debarking, low water temperatures, different water degassing techniques, and varied concentrations of calcium, potassium, magnesium, and copper salts in aqueous measurement solutions. Stable measurements of k(max) were observed at concentrations of calcium, potassium, and magnesium salts high enough to suppress bubble coalescence, as well as with deionized water that was degassed using a membrane contactor under strong vacuum. Bubble formation and coalescence under positive pressure in the xylem therefore appear to be the main cause for declining k(max) values. Our findings suggest that degassing of water is essential for achieving stable and precise measurements of k(max) through woody plant samples. For complete rehydration of woody samples, incubation in water under vacuum for 24 h is suggested as a reliable technique that avoids bubble problems associated with flushing under high positive pressure.