Gold perfusion experiments support the multi‐layered, mesoporous nature of intervessel pit membranes in angiosperm xylem

Summary Fluid transport across intervessel pit membranes of angiosperm xylem plays a major role in plant transpiration, with transport resistance largely depending on pore constriction sizes. Traditionally, fluid particles traversing pit membranes are assumed to cross a single instead of multiple pore constrictions. We tested a multi‐layered pit membrane model in xylem of eight angiosperm species by estimating the size frequency of pore constrictions in relation to pit membrane thickness and compared modelled data with perfusion characteristics of nanoscale gold particles based on transmission electron microscopy. The size frequency of modelled pore constrictions showed similar patterns to the measured number of perfused particle sizes inside pit membranes, although frequency values measured were 10–50 times below modelled data. Small particles enter pit membranes most easily, especially when injected in thin pit membranes. The trapping of gold particles by pore constrictions becomes more likely with increasing pore constriction number and pit membrane thickness. While quantitative differences between modelled and experimental data are due to various practical limitations, their qualitative agreement supports a multi‐layered pit membrane model with multiple pore constrictions. Pore constrictions between 5 and 50 nm are realistic, and confirm the mesoporous nature of pit membranes.