Modified Stereological Method for Analysis of Compound Leaves and an Example of Its Application

Abstract

Stereology, an interdisciplinary field that enables three-dimensional interpretation of planar sections, contains several methods for evaluation of quantitative parameters. The existing leaf sampling procedures were modified and adapted for compound, trifoliate leaves. The similar sampling method could also be applied on other types of compound leaves. Using Trifolium species as model species, we suggested the new way of application of this method in agronomy -precise determination of volume densities of indigestible lignified tissues (which determines forage digestibility), and of chlorenchyma and intercellulars (important for the photosynthesis). Comparison of the results obtained by classic anatomical analysis of leaf cross sections with the results of stereological analysis showed that both methods could be successfully applied for analysis of compact tissues (epidermis, vascular and mechanical). For the tissues that contained intercellulars (palisade and spongy), stereological method gave more precise results, as it enabled separate determination of volume density of intercellulars. A further advantage is that several leaf segments are analyzed and observed from four different angles. The obtained results opened new possibilities of application of stereological method in botanical and agricultural research, especially in prediction of digestibility and nutritional value of forages.

Cytokine associated neopterin response of peripheral blood mononuclear cells to in vitro Epstein–Barr virus transformation process of B lymphocytes

“…Mathematical operations and graphs of the functions were performed with Mathematica 6 (Wolfram 1991), while statistical analysis was performed with Statistica 10. Th ese soft ware packages have been very useful in many other agriculturerelated papers (Babić et al 2011(Babić et al , 2012Dedović et al 2011;Tomic et al 2011;Zorić et al 2011).…”

Stereology, an Unbiased Methodological Approach to Study Plant Anatomy and Cytology: Past, Present and Future

“…Using this approach, the volume density of individual tissues in a leaf was measured by Edwards et al (1999), Klich (2000), Luković et al (2001), Bray and Reid (2002), Luković (2006), Marin et al (2006), Zorić et al (2011;, Moura and Alves (2014), Bernardo et al (2017), and Bertel et al (2017). The proportion of intercellular spaces in mesophyll was also investigated by Albrechtová and Kubínová (1991), Kukkola et al (2005), Albrechtová et al (2007), Lhotáková et al (2008), Psaras (2011), andZorić et al (2011;. The proportion of intercellular spaces in the palisade parenchyma was measured by Konoplyova et al (2008) from micrographs of paradermal sections of the leaves.…”