Abstract
PremisePressure–volume curves are a widely used analytical framework to derive several key physiological traits related to plant–water relations, including a species’ turgor loss point, osmotic potential at full turgor, and the elasticity of cell walls. We developed a novel protocol, including the preparation and treatment of fern gametophytes, to generate data for pressure–volume curve analyses using thermocouple psychrometry.Methods and ResultsGametophytes of the fern species Polystichum lemmonii were grown from spore, harvested, and subjected to a series of drying intervals. We constructed pressure–volume curves using thermocouple psychrometers to calculate gametophyte water potential and a balance to measure relative water loss.ConclusionsWe present the first protocol for fern gametophyte pressure–volume curves that can accurately determine key physiological traits in fern gametophytes such as the turgor loss point and osmotic potential at full turgor.
Highlights
PREMISE: Pressure–volume curves are a widely used analytical framework to derive several key physiological traits related to plant–water relations, including a species’ turgor loss point, osmotic potential at full turgor, and the elasticity of cell walls
We present the first protocol for fern gametophyte pressure–volume curves that can accurately determine key physiological traits in fern gametophytes such as the turgor loss point and osmotic potential at full turgor
One of the most useful and common approaches in plant–water relations research is the pressure–volume curve (PV curve), which describes the relationship between decreasing water potential (Ψ) and relative water content (RWC)
Summary
Spore samples are collected from three mature fern sporophytes of Polystichum lemmonii Underw. at Umatilla National Forest in Oregon, USA (44.783°N, 118.625°W), and Wenatchee National Forest in Washington, USA (47.411°N, 120.908°W). Be sure to have prepared several new Petri dishes with moist filter paper prior to the step; each gametophyte cluster is subjected to a vigorous rinse with deionized water for ca. Gametophyte clusters are transferred to a second Petri dish with moistened filter paper and repeated (we recommend at least three rinses). The gametophyte cluster (hereafter sample) is transferred to dry filter paper and excess water is blotted with Kimwipes (Kimberly-Clark, Irving, Texas, USA). We recommend performing this step under a dissecting microscope to ensure that all excess water is removed. Physiological parameters (e.g., Ψtlp, πo, ε) are derived from the relationship between water potential and relative water content data (e.g., Fig. 1) within the PV curve analytical framework following Tyree and Hammel (1972)
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