The Brazilian Cerrado, a Neotropical savanna, is a fire-prone ecosystem where the ground layer biomass consists mainly of graminoids. However, as for other savannas, the effects of fire cues (such as smoke) on Cerrado grasses do not present a clear pattern, either for germination or seedling development. Smoke can stimulate different stages of the plant life cycle, which can alter the community and invasion processes. So far, most research on the subject focuses on germination, not addressing post-germinative phases, a sensitive stage of plant development. Here, we investigated the effect of smoke on a native (Echinolaena inflexa) and an invasive (Urochloa decumbens) grass species common in the Cerrado. We analysed germinative parameters and seedling mass and length after exposing the seeds to dry smoke for 5, 10, 15 or 20 min. Seedling development was assessed by measuring shoot and root systems after cultivating germinated seeds for 3, 7 or 15 d. Smoke did not affect germination percentages. However, fumigation reduced the mean germination time of both species and the germination onset of E. inflexa. U. decumbens had higher length values in all periods of cultivation, whereas mass values only surpassed that of E. inflexa at 15 d. Smoke exposure reduced the aboveground length of 7-d seedlings of U. decumbens, and mass of 15-d plants of both species. Also, smoke enhanced the root investment of the native and invasive species in different cultivation periods. Therefore, studying post-germinative parameters on seedling development may bring further insights into the smoke effects.

Fire stimulates the germination of most seeds in fire-prone vegetation. Fruits of Leucadendron (Proteaceae) are winged achenes or nutlets that correlate with their requirements for smoke and/or heat in promoting germination. We describe five possible smoke–heat dormancy-release/germination syndromes among plants, of which Leucadendron displays three (no response, smoke only, smoke and heat). As seed-coat thickness varies with seed-storage location (plant or soil) and morphology (winged or wingless), we tested its possible role in water uptake and germination. Species with winged seeds achieved 100% germination in the absence of smoke/heat, seed coats were an order of magnitude thinner, and their permeability greatly exceeded that of nutlets. As seed-coat thickness increased (1) imbibitional water uptake declined at a decreasing rate, and (2) the response to smoke, and to a lesser extent heat, increased linearly to reach levels of germination approaching those of winged seeds. For species responsive to smoke and heat, there was no additive effect when applied together, suggesting that they may have promoted the same physiological process. We conclude that seed-coat thickness holds the key to germination requirements in this genus, independent of seed-storage location or morphology. By what mechanisms (1) the smoke response is greater the thicker the seed coat and (2) smoke chemicals might increase water permeability to explain the non-additive effect of smoke and heat, warrant further investigation.

Smoke-derived seed germination is an important trait for plants to colonize postfire habitats. The well-characterized smoke-derived chemicals of karrikins germinate seeds of species not known to occur after fires in nature. Hence, the ecologically relevant germination cues in smoke remain to be explored for native postfire plants. With the fire-chaser, Nicotiana attenuata, we revisit a bioassay-driven fractionation of liquid smoke to identify ecologically relevant germination cues. By combining bioassay-guided fractionation and comparative unbiased metabolomics, we developed a robust and efficient method to identify germination cues in smoke. Syringaldehyde (SAL) was re-identified as a germination cue in fractions of liquid smoke that promote seed germination. SAL was found to be produced during wildfires in the plant's native habitat, efficiently adsorbed to N. attenuata seeds from aqueous solutions and not readily leached from soil and accurately predicted the boundaries of natural fire events that reflect the occurrence of native postfire N. attenuata populations. We propose that SAL is an ecologically relevant germination cue in smoke for this species.

Fire and grazing are large-scale disturbances that shape the structure and function of open habitats. In temperate grasslands of southern South America, fire is used as a management tool to control tussock grasses and improve forage quality. In this study, we examined if fire and two of its components (heat and smoke) affect germination from the soil seed bank of a temperate grassland in Uruguay. Soil samples were extracted from a recently burned site and from an adjacent area that had not been burned for at least 4 years. The latter was subjected to four treatments: (1) heat shock, (2) smoke, (3) heat shock and smoke and (4) control. The samples were placed in a germination chamber and germination was recorded for 140 days. Field burn was the treatment that differed most from the control. This treatment produced a significant increase in density and richness of germinants and the germination peak preceded those of the remaining treatments. The three treatments involving fire-related cues did not affect the seedling richness and density, but the germination of some individual species was enhanced by some of them, mainly those in which the seeds were smoked. Our results show that fire and its components stimulate the germination of some species of the Río de la Plata grasslands, contrary to what had been observed previously in the region. We also suggest that, unlike Mediterranean-type systems, other fire cues, alone or in combination with heat and smoke, may promote germination after a fire event.

The flora of Mediterranean ecosystems contains families with species having fully and under-developed embryos in their seeds. After-ripening for physiological dormancy release and smoke influence germination in many species. We investigated how after-ripening and embryo growth interact with smoke to influence the temporal dynamics of seedling emergence among fire ephemerals. Seeds were placed in the field and under standardized (50% relative humidity, 30°C) laboratory conditions to test the effects of summer conditions on physiological dormancy loss. Germination was tested with water or smoke compounds (smoke water, KAR1) at a simulated autumn/winter temperature (18/7°C). The timing and amount of seedling emergence with smoke was observed for seeds exposed to near-natural conditions. During summer, physiological dormancy was broken in all species, enabling germination at autumn/winter but not summer temperatures; no embryo growth occurred in seeds with under-developed embryos. At the start of the wet season, seedling emergence from seeds with fully developed embryos occurred earlier than from seeds with under-developed embryos. In a non-consistent manner among our study species, smoke and smoke compounds influenced the rate of embryo growth and amount of germination. Effects of smoke were noticeable in terms of number of emergents in the first emergence season. Among ecologically similar species, we have shown (1) that both thermal and embryo traits exclude germination in the summer, (2) how embryo size influences the timing of seedling emergence in autumn–winter, and (3) a reduced requirement for smoke in the second emergence season after a fire with a shift to reliance on seasonal cues for emergence.

In Mediterranean fire-prone ecosystems, annual species specific to post-fire habitats should have a soil seed bank and should be able to germinate after a fire. Therefore, various fire-related cues can be expected to stimulate germination in post-fire annuals. Germination patterns of the rare annual Chaenorhinum rubrifolium (Plantaginaceae) were examined in response to mechanical scarification, heat shock, aqueous smoke, nitrogenous compounds, gibberellic acid, karrikinolide (KAR1), and mandelonitrile (a cyanohydrin analogue, MAN) under dark and photoperiod conditions in the laboratory. Combinations of these treatments were also included in the experiment. Strong physiological dormancy in the seeds of C. rubrifolium was partially broken by several fire-related germination cues, including smoke and nitrate, under light conditions. KAR1 and MAN also stimulated germination, and the highest improvement in germination was achieved in the KAR1 treatment in the presence of light. Heat shock + smoke and KAR1 + MAN combinations had positive synergetic and additive effects on germination under light conditions, respectively. The light played a crucial role in the promotion of germination. The results suggest that multiple fire-related cues operate to stimulate germination in C. rubrifolium, an annual species from the Mediterranean Basin. However, the species may have a broader germination niche than a fire-restricted one.

The role of smoke in fire-stimulated germination in the Mediterranean Basin has often been underestimated. A few records on smoke-enhanced germination are present in Mediterranean Lamiaceae species, but there is still a shortage of information to allow generalizations about this family to be made. To test the hypothesis that smoke enhances germination in Mediterranean Lamiaceae species, we performed a germination experiment, including aqueous smoke treatments in various concentrations (1:1, 1:10 and 1:100) on seven eastern Mediterranean Lamiaceae taxa. Six of the studied taxa (Lavandula stoechas, Origanum onites, Phlomis bourgaei, Stachys cretica ssp. smyrnaea, Satureja thymbra, Teucrium lamiifolium ssp. stachyophyllum) showed significant increments in germination percentage in at least one smoke treatment, as compared to the control. Moreover, L. stoechas, S. thymbra and T. lamiifolium ssp. stachyophyllum displayed faster germination in at least one smoke treatment than in the control. Of the species showing significant increments in germination percentages after aqueous smoke application, at least one single concentration of smoke solution did not stimulate germination, except in L. stoechas and S. thymbra which responded positively to all smoke treatments. Therefore, the concentration of aqueous smoke that improved germination was species-specific. Our results contribute to the current limited knowledge on smoke-enhanced germination in Mediterranean Lamiaceae, and support the idea that smoke is an important germination cue for this family.

Consider the following four quotations concerning the distinction between breaking dormancy and stimulating germination.

Aerosol smoke and smoke-water can break dormancy and promote seed germination of many plant species. In this study we investigated changes in gene expression after imbibition of light-sensitive Lactuca sativa L. cv. ‘Grand Rapids’ achenes with dilute smoke-water compared to water control samples kept in the dark or continuous light, using the fluorescent differential display technique. Although no difference was detected in the smoke-water versus water control samples germinated in light, smoke-water treatment resulted in the differential display of several expressed sequence tags (ESTs) when compared to water control samples kept in the dark. The most pronounced fragments isolated correspond to known genes related to germination, with functions in cell wall expansion, regulation of translation, the cell division cycle, carbohydrate metabolism and abscisic acid (ABA) regulation. Real-time polymerase chain reaction (PCR) validation revealed that the transcript abundance of the genes, HVA22, short-chain dehydrogenase/reductase and late embryogenesis abundant protein, are upregulated after smoke treatment when compared to control achenes kept in the light. The results indicate that smoke has a dual effect. On the one hand, the smoke can induce genes that may be linked to ABA action, whereas, on the other hand, it elicits a faster germination rate by inducing a similar pattern in gene expression as light treatment. Smoke effects could be manifested mainly through the induction of the cell division cycle, cell wall extension and storage mobilization.

The stimulatory role of 3-methyl-2H-furo[2,3-c]pyran-2-one, a smoke-derived butenolide, on germination and post-germination events is well documented. Previous studies have involved germinating seeds in the continuous presence of the compound. However, commercial growers cannot exploit the potential of the butenolide for large-scale production of crops due to limited availability and environmental constraints. The present study was undertaken to assess the potential of the butenolide as a priming agent of tomato (Solanum esculentum Mill.) seeds. Flow cytometry data revealed that butenolide-primed seeds had a higher percentage of nuclei at the 4C stage than water-primed seeds. Emergence of the radicle was much faster in the primed seeds. After 36 h of imbibition, a higher percentage of the butenolide-primed seeds (22%) exhibited radicle emergence compared to the water-primed seeds (12%). While butenolide-primed seeds initially germinated more rapidly than either water-primed or unprimed seeds in a 48-h period, water-imbibed seeds reached a similar germination level as the butenolide-primed seeds by 60 h of incubation. The butenolide-primed seeds produced significantly (P ≤ 0.05) more vigorous seedlings than water-primed seeds or seeds in the continuous presence of either water or butenolide. A gradual decrease in the seedling vigour index was recorded for both water and butenolide-primed seeds with increased seed storage at room temperature. Nevertheless, the vigour index was significantly greater in the butenolide-primed seeds than the water-primed seeds. Vigour indices were significantly (P ≤ 0.05) higher for the butenolide-primed seeds under various stress conditions (salinity, osmoticum or temperature) compared to control or water-primed seeds. Results of the present study suggest that the butenolide is a good seed-priming agent. Additionally, primed seeds retained the promotive effect for a considerable time. This was also the case for tomato seeds under various simulated field stress conditions.

Nitrates have long been known to promote seed germination in many species, and various proposals have been made regarding the role of nitrates in this process. One hypothesis suggests a possible ecological role for nitrate, providing the seeds with a report of soil nitrogen status and the proximity of already established plants. However, nitrogen oxides (NO, NO2), which are present in the soil, also may be information carriers that indicate not only soil nitrate content, but also microbial activity and, therefore, soil quality. Because of annual variation of soil trace gas fluxes, seeds could be provided with information on seasonal and climate changes in their surroundings. Thus, nitrogen oxides would be the outer information carriers providing the seeds with integral data about many important factors required for successful germination and seedling establishment.

This study investigated the effects of 3-methyl-2H-furo[2,3-c]pyran-2-one, a germination active butenolide present in plant-derived smoke, gibberellic acid and smoke water on seeds of Australian Asteraceae exposed to different light regimes. Seeds of all species required light, with maximum germination occurring under white light, or light dominated by 640 nm. Compared to untreated seeds, butenolide increased germination of Angianthus tomentosus, Gnephosis tenuissima, Myriocephalus guerinae, Podolepis canescens and Rhodanthe citrina at suboptimal light wavelengths and in the dark to a level equal to, or greater than, smoke water. Germination of Erymophyllum glossanthus and Gnephosis acicularis was not promoted by butenolide or smoke water under any light regime. The action of gibberellic acid was compared to that of butenolide for three species (Angianthus tomentosus, Myriocephalus guerinae and Podolepis canescens), and both compounds were found to stimulate germination. This study provides evidence that butenolide can act in a similar fashion as gibberellic acid in promoting seed germination of light-sensitive seeds. The ecological significance of these findings is discussed.

The native tobacco, Nicotiana attenuata, synchronizes its germination with the immediate post-fire environment with a combination of germination stimulants found in wood smoke and inhibitors from the unburned litter of the dominant vegetation. The inhibitors override the stimulants and prevent seeds from germinating maladaptively in unburned habitats adjacent to burns. To understand the physiological basis of this environmental control of germination, we tested several previously isolated signals, phytohormones and their respective biosynthesis inhibitors. The germination inhibitors methyl jasmonate (MeJA, a constituent of sagebrush litter), bornane-2,5-dione (BD, a constituent of juniper litter extract, JLE) and JLE did not alter abscisic acid (ABA) content of imbibed seeds. Treatment with the ABA biosynthesis inhibitor, fluridone, inhibited the dormancy-inducing effects of BD, JLE and MeJA, but surprisingly did not affect endogenous ABA levels in treated seeds. However, ABA leached from litter of the species, which dominate the plant community before fires, plays an important role in germination control. We conclude that N. attenuata seeds, which can lie dormant in the soil for 150 years between fires, time their germination with the post-fire environment by responding to smoke, ABA and four terpenes (BD, 1,8-cineole, β-thujaplicin and camphor) leaching from the litter of the dominant vegetation.

Exposure to smoke is required for the germination of seeds from dormant genotypes of Nicotiana attenuata, a post-fire annual of the Great Basin Desert. Germination can be elicited by GA1,3,4,7 treatments and inhibited by the GA biosynthesis inhibitor, paclobutrazol (PAC), abscisic acid (ABA) and terpenes leached from unburned litter of the plant’s natural habitat. We analysed the endogenous GA and ABA dynamics during the 22 h after imbibition, when smoke-treated dormant seeds commit to germination. Extractable GA1+3 pools decreased in all seeds, but the decrease was more dramatic within 2 h of smoke exposure, which was followed by an increase between hours 2 and 4. Extractable ABA pools increased shortly after imbibition and remained stable in control, water-treated seeds, but decreased sharply in smoke-treated seeds. PAC completely inhibited smoke-induced germination when seeds were treated for 12 h after smoke exposure, consistent with the requirement of de novo GA synthesis for germination. Smoke treatment in the dark did not result in germination, whereas GA3 treatment did, a result consistent with phytochrome-mediated GA biosynthesis. Smoke exposure dramatically increased the sensitivity of seeds to exogenous GA3 treatments in both the light and dark, and light exposure increased this sensitivity an additional tenfold. Taken together, these results suggest that while germination requires endogenous GA synthesis, the effects of smoke treatment increase GA sensitivity, which is correlated with a decrease in endogenous ABA pools.

Patterns of dormancy and smoke responsiveness in artificially buried seeds were investigated in eight native plant species from the species-rich mediterranean-type climate zone of south-western Australia. A comparison was made between germination and viability behaviour of shelf- and field-soil-stored seed, with and without smoke treatment, at least every 3 months. These comparisons corresponded with each of the four seasons. The species chosen germinated with the aid of smoke under field or glasshouse conditions (termed ‘dormant’; n = 4) or produced low and variable germination under glasshouse conditions with smoke (termed ‘deeply dormant’; n = 4). Three trends were observed in viability of soil-stored seeds over 450 d: no decline, gradual decline or late-onset decline. In addition, various germination responses to soil burial and aerosol smoke were observed. Burial was not required for optimal germination in Anigozanthos manglesii. However, for all other species tested, maximum germination was observed only following a period of burial. This was manifested in a germination response without smoke after a short period of burial (Stylidium affine and Conospermum triplinervium) or a longer period of burial (Conostylis neocymosa, Hibbertia commutata, Leucopogon conostephioides, Stirlingia latifolia and Stylidium crossocephalum). Smoke treatment led to high germination in buried seed of S. affine, S. crossocephalum and H. commutata. The patterns of germination detected in this limited number of species indicate that a variety of mechanisms may exist, both temporally and spatially, in south-western Australian species, and support further research of this type for horticulture and land restoration.