DOI: 10.5061/dryad.x95x69phv
title: Leaf Anatomical Properties

description: Prunus L. subgen. Cerasus Mill. (Rosaceae) contains numerous economically
 important species, including fruit trees and ornamental plants. This study
 compares the leaf anatomy and calcium oxalate crystal types of 98
 individuals of 14 Prunus subgen. Cerasus taxa from Turkey. We stained leaf
 sections with Safranin-Alcian Blue double stain and mapped calcium oxalate
 crystals under light microscope with polarized filter. We used one-way
 ANOVA to determine the statistical significance of quantitative data.
 Significant quantitative and qualitative data were analyzed using PCoA to
 establish relationships among taxa. We observed three different crystal
 distribution patterns and three crystal types, finding druses present in
 all taxa, while some taxa also have sand or prismatic forms. Most
 characters showed high variation. Our analysis shows that vascular bundle,
 midrib, upper epidermis and lower epidermis width/length ratios; midrib
 epidermis, upper epidermis and phloem cell lengths; upper epidermis and
 lower epidermis cell widths and lower cuticle thickness are not
 taxonomically useful, while features of the bundle sheath, epidermis cell
 surface, phloem fibers, mesophyll layers and angle of blade between the
 midrib can be used in conjunction with other characters to distinguish
 certain taxa. Overall, PCoA delineated two rough groups, separating P.
 avium, P. cerasus and P. mahaleb from the rest of the studied taxa. In
 investigating the anatomical variation between different individuals of
 these taxa, we found that some features previously used in infraspecific
 taxa, such as trichomes, varied sometimes even within an individual,
 casting doubt on the wisdom of using these for systematic work in subgenus
 Cerasus.
description: <i>Plant Material</i> Subgenus
 <i>Cerasus</i> in Turkey consists of deciduous shrubs or
 small- to medium-sized trees which grow primarily on open hillsides. We
 selected at random mature sun leaves of 14 taxa from subgenus
 <i>Cerasus</i> of the genus<i> Prunus</i>,
 including the infraspecific taxa used by Browicz (1972) (<i>P.
 prostrata</i> var. <i>glabrifolia</i>, <i>P.
 prostrata</i> var. <i>prostrata</i>, <i>P.
 microcarpa</i> subsp.<i> microcarpa</i>, <i>P.
 microcarpa</i> subsp. <i>tortuosa</i>, <i>P.
 avium</i>,<i> P. cerasus</i>, <i>P.
 mahaleb</i> var. <i>mahaleb</i>, <i>P.
 mahaleb</i> var. <i>alpina</i>, <i>P.
 brachypetala</i>,<i> P. hippophaeoides</i>, <i>P.
 angustifolia</i> var. <i>angustifolia</i>, <i>P.
 angustifolia</i> var. <i>sintenisii</i>, <i>P.
 incana</i> var.<i> incana</i>, <i>P.
 incana</i> var. <i>velutina</i>), which were collected
 throughout Turkey in summers 2017-2019. The voucher specimens are
 deposited in the Istanbul University Faculty of Science Herbarium
 (ISTF).  <i>Anatomical
 Studies</i> Mature leaves were fixed in 10%
 glycerin solution in 70% ethanol. The fixed leaves were then dehydrated
 through ascending alcohol series (70% EtOH, 80% EtOH, 96% EtOH, 100% EtOH,
 Xylol I, Xylol II) and embedded in paraffin blocks. Sections were taken
 with Leica RM2125 manual rotary microtome at 6 micrometer. The sections
 were stained with Safranin O and Alcian Blue double stain after
 deparaffinization (Tolivia, D. and Tolivia 1987). Leaf material for
 crystal examinations was prepared according to the method used by Lersten
 and Horner (2000). Briefly, the leaves were cut into small pieces and
 bleached using sodium hypochlorite (commercial bleach). Leaves were rinsed
 with distilled water, mounted with glycerin solution and examined under
 polarized filter. All slides were examined with ZEISS Axio Scope A1
 research microscope and measurements were taken with Argenit Kameram v3.1
 Software.
description: An explanation sheet is added to the excel file.