Pine tree substrates (PTSs) may provide growers with sustainable substrate component options. Improved processing of PTS components has provided new materials with little scientific evaluation or understanding of their hydrophysical behavior and properties. Moisture retention characteristics were developed for two PTSs and four traditional greenhouse components: sphagnum peat, coconut coir, perlite, pine bark, shredded-pine-wood (SPW), and pine-wood-chips (PWC). Mixtures of peat containing 10%, 20%, 30%, 40%, and 50% of perlite, SPW, or PWC were also characterized. Hydrophysical properties were measured, allowing for comparison of the PTS components to the more traditional substrate components (peat, coir, perlite, and pine bark). The SPW was constructed to retain water similarly to peat and pine bark, whereas the PWC was made to increase drainage like perlite. Shredded pine wood had higher total porosity and more easily available water than did PWC components. Total porosities of SPW and PWC were similar to pine bark and coir; air space and drainage were higher than peat and coir because of the lower percentage of fine particles in the PTS components. The two PTS components had a greater influence on water drainage and retention dynamics than did perlite when amended with peat as an aggregate. Water release patterns of SPW or PWC components at low tensions were lower than peat and greater than pine bark; drainage was similar to perlite at higher tensions. Equilibrium capacity variable models predicted similar physical properties (and trends) across multiple container sizes for peat mixes amended with perlite, SPW, or PWC. The impact of PWC on drainage and aeration was similar to perlite in all containers, but these effects were greater in smaller containers.