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?(Fig.3).3). This observation corresponded to the change in the opening index of the petals and stamens, which both began to extend gradually GDC941 at 10:00 h, reached a peak at 12:00 h and then decreased to nearly zero again at about 16:00 h (Fig. ?(Fig.3).3). The movement pattern of petals was similar on the first and second days. However, for stamens, the opening index of the second-day flower reached a lower plateau between 11:00 and 15:00 h rather than a peak as in the first-day flower (Fig. ?(Fig.33). Figure 3. Diagram of the petal opening index (A, mean �� SE) and the stamen opening index (B, mean �� SE) on the first day (open circles) and second day (solid circles) in P. hexandrum flowers. The time interval is 30 min. Stigmatic pollen load The pollen grain counts on the stigma increased significantly during two different periods of anthesis. Pollen deposition was low in flowers on the first day of opening (18.9 �� 7.5 grains), but it increased significantly after flowers had experienced one night's closure (247.1 �� 79.1, Mann�CWhitney U test, Z = ?3.16, P selleck screening library pollen deposition on diglyceride stigmas. These results indicated that self-pollination occurred during the period of flower closure. Figure 4. Stigmatic pollen load (mean �� SE) on three consecutive days. Measurement was at 17:00 h on the first day, 9:00 h on the second day, 17:00 h on the second day and 9:00 h on the third day. Significant differences between means are indicated by different ... Discussion Our pollination treatments confirmed that Himalayan mayapple is SC and capable of autogamy. Our observations of flower behaviours and manipulations indicated that autogamy in P. hexandrum was facilitated by petal closure, rather than by pistil movement (Ma et al. 1997; Xu et al. 1997). The pistil was indeed inclined at flower opening, but it did not move towards the anthers during anthesis. Furthermore, we observed that bees were potential effective pollinators for this early flowering herb in the alpine area. Wind pollination is unlikely in the species, given that the showy flowers release sticky pollen in tetrads. Self- and cross-pollination yielded similar seed production, suggesting no pre-seedling inbreeding depression. Automatic self-pollination occurred when the petals were closed, indicating that it was delayed selfing rather than prior selfing as previously thought.