Bipolar pollen germination in blue spruce (Picea pungens).

Direct growth of a pollen tube is an effective mechanism of sperm delivery characteristic for the majority of seed plants. In most cases, only one tube grows from one grain to perform the delivery function; meanwhile in Picea the appearance of two tubes from a single pollen grain is quite common during in vitro germination. Here, we describe the phenomenon of bipolar germination and test two hypotheses on its nature and possible role in gametophyte functioning. The hypothesis on "trophic" function of multiple tubes provoked by poor nutrition discussed in literature was not confirmed by in vitro growth tests; bipolar germination strongly decreased with lowering sucrose availability. The highest proportion of bipolar germination occurred in optimal conditions. We then assumed that bipolar germination occurs because turgor pressure is a non-directional force and effective systems of cell wall mechanical regulation are lacking. In hypertonic medium, bipolar germination was sufficiently lower than in isotonic medium, which was consistent with prediction of the «mechanical» hypothesis. Scanning electron microscopy and fluorescence microscopy analysis of pollen morphology and cell wall dynamics during both types of germination showed that the appearance of a single tube or bipolar germination depends on the extension of exine rupture. Cell wall softening by short-term ·OH treatment sufficiently decreased the percent of bipolar germination without affecting total germination efficiency. We concluded that mechanical properties of the cell wall and turgor pressure could shift the balance towards one of the germination patterns.