Starch granules from higher plants contain alternating zones of semicrystalline and amorphous material known as growth rings. The regulation of growth ring formation is not understood. We provide several independent lines of evidence that growth ring formation in the starch granules of potato (Solanum tuberosum) tubers is not under diurnal control. Ring formation is not abolished by growth in constant conditions, and ring periodicity and appearance are relatively unaffected by a change from a 24-h to a 40-h photoperiod, and by alterations in substrate supply to the tuber that are known to affect the diurnal pattern of tuber starch synthesis. Some, but not all, of the features of ring formation are consistent with the involvement of a circadian rhythm. Such a rhythm might operate by changing the relative activities of starch-synthesizing enzymes: Growth ring formation is disrupted in tubers with reduced activity of a major isoform of starch synthase. We suggest that physical as well as biological mechanisms may contribute to the control of ring formation, and that a complex interplay of several factors may by involved.Starch granules from every higher plant species studied so far contain alternating regions of semicrystalline and amorphous material commonly known as growth rings. Growth rings can be observed by light microscopy, by atomic force microscopy, and by scanning and transmission electron microscopy (SEM and TEM) after treatment of granules with acid or degradative enzymes. These methods reveal that the rings represent alternating concentric layers of high/low refractive index, density, crystallinity, and resistance to chemical and enzymatic attack (Badenhuizen, 1939; Badenhuizen, 1959; Buttrose, 1960;Gallant and Guilbot, 1969;Hall and Sayre, 1973; Baker et al., 2001).The origin of growth rings remains obscure. Previous studies have suggested that one of two biological mechanisms could regulate their formation. First, their formation could be under the control of a diurnal rhythm that is dependent on day/night variations in the environment, such as a light/dark regime or alternating temperature cycles. Meyer (1895) hypothesized that growth of the granule follows a diurnal rhythm and that one growth ring is laid down per day. Support for this comes from studies of growth rings in the starch of developing cereal endosperm. Granules from barley (Hordeum vulgare) endosperm were claimed to have one growth ring for each day after their initiation (Buttrose, 1960), and growth rings were not visible in granules from the endosperm of wheat (Triticum aestivum) and barley plants grown in constant light and temperature (Van de Sande-Bakhuyzen, 1925; Buttrose, 1960 Buttrose, , 1962. Rings reappeared in the peripheral regions of granules when plants grown initially in constant conditions were transferred to a day-night regime during the course of granule development (Buttrose, 1962). These observations lead Buttrose (1962) to propose that growth ring formation is controlled by a diurnal rhythm that is dependent on ...