Development of the calcium oxalate crystal macropattern in pomegranate (Punica granatum, Punicaceae)

Oxalate crystals are very common in angiosperms, but few descriptions of their macropattern (crystal types, their tissue distribution, and development) exist. Because unusually large prismatic crystals and druses, are known from pomegranate, we traced the development of crystal macropattern in various-aged leaf samples from a living plant and from herbarium specimens using unstained whole mounts (some bleached and cleared), stained leaf samples, and leaf and stem cross sections. Preparations were viewed with bright-field light microscopy and with crossed polarizers. Prismatics appear first in the subapical mid-mesophyll layer of a leaf 650 [micro]m long. Additional prismatics form basipetally in the enlarging lamina. A preemptive wave of small prismatics appears basipetally in the midrib. Druses form secondarily acropetally in petiole and midrib, while existing lamina prismatics enlarge and new ones develop among them in mid-mesophyll. Prismatics produced early expand vertically, and many eventually extend from epidermis to epidermis. Later-formed prismatics attain intermediate sizes. No crystals form along lamina veins, but in older leaves, druses occur in spongy mesophyll, mostly near major vein junctions. In the stem, druses are restricted to phloem fibers. No phloem fibers occur in the leaf trace or midvein; therefore, petiolar and midrib druses are only in parenchyma, not in phloem.