Up in smoke: I. Smoke-derived germination cues for postfire annual,Nicotiana attenuata torr. Ex. Watson

Abstract

Some postfire annuals with dormant seeds use heat or chemical cues from charred wood to synchronize their germination with the postfire environment. We report that wood smoke and polar extracts of wood smoke, but not the ash of burned wood, contain potent cue(s) that stimulate germination in the postfire annual plant,Nicotiana attenuata. We examined the responses of seeds from six populations of plants from southwest Utah to extracts of smoke and found the proportion of viable seeds that germinated in the presence of smoke cues to vary between populations but to be consistent between generations. With the most dormant genotypes, we examine three mechanisms by which smoke-derived chemical cues may stimulate germination (chemical scarification of the seed coat and nutritive- and signal-mediated stimulation of germination) and report that the response is consistent with the signal-mediated mechanism. The germination cue(s) found in smoke are produced by the burning of hay, hardwood branches, leaves, and, to a lesser degree, cellulose. Moreover, the cues are found in the common food condiment, “liquid smoke,” and we find no significant differences between brands. With a bioassay-driven fractionation of liquid smoke, we identified 71 compounds in active fractions by GC-MS and AA spectrometry. However, when these compounds were tested in pure form or in combinations that mimicked the composition of active fractions over a range of concentrations, they failed to stimulate germination to the same degree that smoke fractions did. Moreover, enzymatic oxidation of some of these compounds also failed to stimulate germination. In addition, we tested 43 additional compounds also reported from smoke, 85 compounds that were structurally similar to those reported from smoke and 34 compounds reported to influence germination in other species. Of the 233 compounds tested, 16 proved to inhibit germination at the concentrations tested, and none reproduced the activity of wood smoke. By thermally desorbing smoke produced by cellulose combustions that was trapped on Chromosorb 101, we demonstrate that the cue is desorbed between 125 and 150°C. We estimate that the germination cues are active at concentrations of less than 1 pg/seed and, due to their chromatographic behavior, infer that a number of different chemical structures are active. In separate experiments, we demonstrate that cues remain active for at least 53 days in soil under greenhouse conditions and that the application of aqucous extracts of smoke to soil containing seeds results in dramatic increases in germination of artificial seed banks. Hence, although the chemical nature of the germination cue remains elusive, the stability of the germination cues, their water-solubility, and their activity in low concentrations suggest that these cues could serve as powerful tools for the examination of dormant seed banks and the selective factors thought to be important in the evolution of postfire plant communities.