Teasing out elevational trends in infraspecific Prunus taxa: A vein analysis approach

Using 33 specimens collected from across their range in Turkey, we demonstrate that the subspecies of Prunus microcarpa C.A.Mey react very differently to altitude. We first outline a simplified, flexible protocol for sectioning and removing the epidermis of small, difficult-to-image leaves for leaf vein studies. We then used venation analysis software to evaluate the two subspecies of this wild cherry in relation to altitude. We also found key differences in venation features between short-shoot and long-shoot leaves for each taxon. Differences include statistically significant negative correlation between vein density, and positive correlation between areole area and altitude in long-shoot leaves of Prunus microcarpa subsp. microcarpa, while its short-shoot leaves had a positive relationship between maximum areole area, and negative relationship between vein density, numbers of veins and endpoints. Meanwhile, P. microcarpa subsp. tortuosa (Boiss. & Hausskn.) Browicz recorded trends that were largely opposite of this, but beside vein thickness and areole area, were not statistically significant. This difference may be part of each taxon's overarching syndrome of anatomical and morphological adaptations to its external environment. RESEARCH HIGHLIGHTS: Features of vein density and thickness fell, while areole area and vein length rose with altitude in P. microcarpa. P. microcarpa subsp. tortuosa showed opposite trends, but reacted less strongly to altitude. Short-shoot and long-shoot have significantly different venation parameters. Using sections proportionate to leaf size is useful to compare venation of leaves that vary due to dimorphism. We discuss protocol strategies for imaging of difficult leaves for venation analyses.