Anatomical and Biochemical Traits Related to Blue Leaf Coloration of Selaginella uncinata

Selaginella uncinata shows particularly rare blue leaves. Previous research has shown that structural interference by the cell wall of adaxial epidermal cells imparts blue coloration in leaves of S. uncinata; the objective of this study was to see whether anthocyanins might additionally contribute to this color, as changes in pH, and conjugation with metals and other flavonoids is also known to result in blue coloration in plants. We compared anatomical and biochemical traits of shade-grown (blue) S. uncinata leaves to high light (red) leaves of the same species and also to a non-blue (green) leaves of a congeneric S. kraussiana. By examining the anatomical structure, we found that the shape of adaxial epidermis of S. uncinata leaves was convex or lens-shaped on the lateral view and irregular circles with smooth embossment on the top view. These features were different from those of the abaxial and adaxial epidermis of S. kraussiana. We suspect that these structures increase the proportion of incident light entering the cell, deepening the leaf color, and therefore may be related to blue leaf color in S. uncinata. By examining biochemical traits, we found little difference in leaf pH value among the leaf types; all leaves contained several metal ions such as Mg, Fe, Mn, and copigments such as flavones. However, because there was no anthocyanin in blue S. uncinata leaves, we concluded that blue coloration in S. uncinata leaves is not caused by the three hypotheses of blue coloration: alkalization of the vacuole pH, metal chelation, or copigmentation with anthocyanins, but it may be related to the shape of the leaf adaxial epidermis.