Measuring tissue water potential in marine macroalgae via an updated Chardakov method

Regulation of tissue water potential is a key mechanism in macroalgal osmotic responses to changing external osmotic conditions, which are common in tidally influenced estuarine and intertidal systems. Nevertheless, significant knowledge gaps exist in our understanding of osmotic responses in macroalgae because few methods measure osmotic potential within macroalgal tissues. Leaf psychrometers have furthered understanding of osmotic potentials in terrestrial plant water relations, yet these have not been developed to measure the range of highly negative potential values found in marine macroalgae. To address these gaps, we present an effective, updated version of the Chardakov method to measure tissue water potential in macroalgae. Here, we present a case study examining macroalgal response in tissue water potential by two morphologically and evolutionarily distinct species, Ulva lactuca (Chlorophyta) and Hypnea musciformis (Rhodophyta) to four paired salinity and nutrient treatments at two temperatures. These treatments simulate a gradient from full coastal ocean conditions to brackish submarine groundwater discharge, an ecosystem type found on basaltic shorelines. Both algae demonstrated plasticity in osmotic response to submarine groundwater discharge with significant positive correlations between tissue water potential and proportion of submarine groundwater discharge in the treatment. These results are the first to describe macroalgal response in tissue water potential, a first step to understanding algal physiological ecology in such complex coastal environments. This revised Chardakov method is a valuable tool to better understand species-specific osmotic responses to ecologically relevant conditions, and can augment the study of other tidal systems and ontogenetic stages. Marine plants occurring in tidally-influenced estuarine nearshore ecosystems experience daily oscillations from fresh to marine salinities. Under the stress of changing external salinities regulation of solute concentrations within plant cells is critical to continued growth and photosynthesis. However, a lack of available methodologies has prevented significant breakthroughs in understanding of water relations in marine plants since the 1980s. This work updates the Chardakov Method, a classic method of measuring tissue water potential. This simple and cost-effective method is applied to examining the regulation of internal cellular solute concentrations for two Hawaiian marine plant species under simulated conditions of nearshore lowered salinity and higher nutrients. This work demonstrates the use of this methodology for measuring marine plant water potentials, and measures salinity-driven regulation of internal cellular solute concentrations for two species. The authors encourage the use of this method for further investigation of water relations in marine plants under oscillating salinities.