When is lethal deceptive pollination maintained? A population dynamics approach.

BACKGROUND AND AIMS: Not all plant-pollinator interactions are mutualistic, and in fact, deceptive pollination systems are widespread in nature. The genus Arisaema has a pollination system known as lethal deceptive pollination, in which plants not only attract pollinating insects without providing any rewards, but also trap them until they die. Many Arisaema species are endangered from various disturbances including reduction in forest habitat, modification of the forest understory owing to increasing deer abundance, and plant theft for horticultural cultivation. We aimed to theoretically investigate how lethal deceptive pollination can be maintained from a demographic perspective and how plant and pollinator populations respond to different types of disturbance. METHODS: We developed and analysed a mathematical model to describe the population dynamics of a deceptive plant species and its victim pollinator. Calibrating the model based on empirical data, we assessed the conditions under which plants and pollinators could coexist, while manipulating relevant key parameters. KEY RESULTS: The model exhibited qualitatively distinct behaviours depending on certain parameters. The plant becomes extinct when it has a low capability for vegetative reproduction and slow transition from male to female, and plant-insect co-extinction occurs especially when the plant is highly attractive to male insects. Increasing deer abundance has both positive and negative effects because of removal of other competitive plants and diminishing pollinators, respectively. Theft for horticultural cultivation can readily threaten plants whether male or female plants are frequently collected. The impact of forest habitat reduction may be limited compared to that of other disturbance types. CONCLUSIONS: Our results have emphasised that the demographic vulnerability of lethal deceptive pollination systems would differ qualitatively from that of general mutualistic pollination systems. It is therefore important to consider the demographics of both victim pollinators and deceptive plants to estimate how endangered Arisaema populations respond to various disturbances.

How specialized is a soil specialist? Early life history responses of a rare Eriogonum to site-level variation in volcanic soils.

PREMISE: Understanding edaphic specialization is crucial for conserving rare plants that may need relocation due to habitat loss. Focusing on Eriogonum crosbyae, a rare soil specialist in the Great Basin of the United States, we asked how site-level variation among volcanic soil outcrops affected plant growth and population distribution. METHODS: We measured emergence, survival, size, and biomass allocation of E. crosbyae seedlings planted in soils collected from 42 outcrops of actual and potential habitat. We also measured phenotypic variation in the wild, documented abiotic and biotic components of E. crosbyae habitat, re-surveyed Nevada populations, and evaluated occupancy changes over time. RESULTS: Plants responded plastically to edaphic variation, growing larger and allocating relatively more to aboveground tissues in soils with greater nutrient availability and growing smaller in soils higher in copper in the field and the greenhouse. However, the chemical and physical soil properties we measured did not predict site occupancy, nor was plant phenotype in the greenhouse different when plants were grown in soils from sites with different occupation status. We observed occupation status reversals at five locations. CONCLUSIONS: Eriogonum crosbyae performed well in soils formed on hydrothermally altered rocks that are inhospitable to many other plants. Extirpation/colonization events observed were consistent with metapopulation dynamics, which may partially explain the patchy distribution of E. crosbyae among outcrops of potential habitat. While soil properties did not predict site occupancy, early life stages showed sensitivity to soil variation, indicating that seedling dynamics may be important to consider for the conservation of this soil specialist.

Small herbaria contribute unique biogeographic records to county, locality, and temporal scales.

PREMISE: With digitization and data sharing initiatives underway over the last 15 years, an important need has been prioritizing specimens to digitize. Because duplicate specimens are shared among herbaria in exchange and gift programs, we investigated the extent to which unique biogeographic data are held in small herbaria vs. these data being redundant with those held by larger institutions. We evaluated the unique specimen contributions that small herbaria make to biogeographic understanding at county, locality, and temporal scales. METHODS: We sampled herbarium specimens of 40 plant taxa from each of eight states of the United States of America in four broad status categories: extremely rare, very rare, common native, and introduced. We gathered geographic information from specimens held by large (≥100,000 specimens) and small (<100,000 specimens) herbaria. We built generalized linear mixed models to assess which features of the collections may best predict unique contributions of herbaria and used an Akaike information criterion-based information-theoretic approach for our model selection to choose the best model for each scale. RESULTS: Small herbaria contributed unique specimens at all scales in proportion with their contribution of specimens to our data set. The best models for all scales were the full models that included the factors of species status and herbarium size when accounting for state as a random variable. CONCLUSIONS: We demonstrated that small herbaria contribute unique information for research. It is clear that unique contributions cannot be predicted based on herbarium size alone. We must prioritize digitization and data sharing from herbaria of all sizes.

Remnants of populations provide effective source material for reintroduction of an endangered Hawaiian plant, Schiedea kaalae (Caryophyllaceae).

PREMISE OF THE STUDY: Reintroductions may be essential to prevent extinction of many critically endangered species. Ideally, reintroduction efforts rely on adjacent source populations, but limited source material may necessitate crossing individuals from different and possibly distant populations. To determine the consequences of integrating multiple populations in reintroductions, we investigated levels of inbreeding depression, outbreeding depression, and heterosis for populations of Schiedea kaalae (Caryophyllaceae), an endangered species endemic to the Wai'anae and Ko'olau Mountains of O'ahu, Hawai'i. The possibility of gene flow among plants was explored through pollinator observations. METHODS: Individuals from ex situ living collections of nine populations were hand-pollinated with pollen from the same plant, plants from the same population (for three populations only), or plants from different populations. Progeny were outplanted into two common gardens, one in each mountain range on O'ahu. Cumulative fitness was estimated using several independent life history stages. KEY RESULTS: Inbreeding depression was minimal, and no outbreeding depression was detected. In contrast, strong heterosis was evident in progeny from between-population crosses, which had higher relative fitness than progeny from self-pollinations or within-population crosses. Observations of floral visitors provided the first evidence that biotic pollination may be important for this species. CONCLUSIONS: Results demonstrate the ability to conduct genetic rescue of rare species and suggest that reintroductions may be most successful using heterotic individuals from crosses between populations and at sites where pollinators are present and promote outcrossing.

Demography and threats to population growth of Cirsium pitcheri, a threatened dune plant, in Wisconsin.

Multi-year and multi-site demographic data for rare plants allow researchers to observe threats and project population growth rates and thus long-term persistence of the species, generating knowledge, which allows for effective conservation planning. Demographic studies across more than a decade are extremely rare but allow for the effects of threats to be observed and assessed within the context of interannual environmental variation. We collected demographic data on the Threatened plant Cirsium pitcheri in two sites from 2011-2022. These sites were chosen because one exhibited the presence of non-native seed predators while the other did not, and we hypothesized that we would see declines and potentially extinction of the population threatened by predation. Over the course of our study, we observed additional threats, such as human trampling and high lake levels, which led to significant erosion, sand burial, and storm damage to plants. We find high interannual variation in vital rates and population growth rates for both populations, which mask the overall effects of predation. We observed dramatic declines in plant survivorship and population growth rates in both sites in the years with high lake levels. We conclude that high lake levels, which are expected to become more frequent with climate change, pose a significant threat to all near-shore populations of C. pitcheri.

Predispersal seed predation is higher in a rare species than in its widespread sympatric congeners (Astragalus, Fabaceae).

PREMISE OF THE STUDY: Rare plants may be rare, in part, because they are more susceptible to damage by predispersal insect seed predators than widespread congeneric species; thus, seed predation may be an important determinant of plant rarity. Scant evidence exists to either support or reject this hypothesis, limiting our ability to predict herbivore effects on plant rarity. We used a comparative framework to test this hypothesis in rare-common, sympatric congeners. METHODS: We compared seed consumption by insect type and seed production among a rare Astragalus (Fabaceae) species and two sympatric, widespread congeners. We measured plant traits and tested whether traits were correlated with seed damage within and among rare-common species. KEY RESULTS: Seed predation was significantly higher in a rare species than in common congeners over 2 yr. Seed production per pod was significantly lower in the rare species. Seed weevils exclusively consumed seeds of the rare species. Higher seed predation in the rare species is related to a combination of factors: plant phenology, dispersal ability, and vigor; seed predator identity; and insect phenology. CONCLUSIONS: Our results support the hypothesis that a rare plant is more susceptible to seed predators than two common, sympatric congeners. Seed predator reduction is a promising management tool to increase seed-set, recruitment, and survival of the rare species. Further studies that compare rare-common, sympatric congeners are greatly needed to broadly evaluate the hypothesis concerning rarity and susceptibility to seed predators and to inform conservation plans for rare species.

Using photogrammetry to create virtual permanent plots in rare and threatened plant communities.

Premise: Many plant communities across the world are undergoing changes due to climate change, human disturbance, and other threats. These community-level changes are often tracked with the use of permanent vegetative plots, but this approach is not always feasible. As an alternative, we propose using photogrammetry, specifically photograph-based digital surface models (DSMs) developed using structure-from-motion, to establish virtual permanent plots in plant communities where the use of permanent structures may not be possible. Methods: In 2021 and 2022, we took iPhone photographs to record species presence in 1-m2 plots distributed across alpine communities in the northeastern United States. We then compared field estimates of percent coverage with coverage estimated using DSMs. Results: Digital surface models can provide effective, minimally invasive, and permanent records of plant species presence and percent coverage, while also allowing managers to mark survey locations virtually for long-term monitoring. We found that percent coverage estimated from DSMs did not differ from field estimates for most species and substrates. Discussion: In order to continue surveying efforts in areas where permanent structures or other surveying methods are not feasible, photogrammetry and structure-from-motion methods can provide a low-cost approach that allows agencies to accurately survey and record sensitive plant communities through time.

Long-term seed bank persistence in a stochastic desert environment.

Seed banks, the collection of viable seeds in the soil, are particularly important determinants of population survival in highly variable environments. Predictions of increased stochasticity in the amount and timing of precipitation in desert environments raise the question of how seed banks of desert species will respond to climate change, and ultimately, whether these species will persist. Here, we present data from our long-term studies of germination requirements and seed bank dynamics in a rare desert gypsophile perennial, Arctomecon californica (Las Vegas bearpoppy). Arctomecon californica is a relatively short-lived plant that recruits from seed in sequences of unusually favorable years. We used germination experiments, an in situ seed bank study, and a 15-year field seed retrieval study to examine factors affecting seed bank persistence. In the germination study, a majority of seeds remained dormant, despite a wide variety of treatments, suggesting that a large proportion of the seed dispersed each year has cue-nonresponsive dormancy. Our in situ seed bank study showed that seed density varied widely between sites, among transects, and among samples within a transect. The patchiness of seeds in the soil highlights the importance of protecting large areas where A. californica populations are known to have existed in the past. The seed retrieval study provided strong evidence that this species has a long-lived seed bank in which only a small fraction of seeds (roughly 5%) become nondormant each year, allowing seed banks of this species to last up to 20 years without a seed production event. Whether this impressive life-history strategy can maintain the species in the face of climate change depends on the future frequency of the well-timed precipitation that allows for the establishment of new cohorts of adult plants.

Population Genomics Study and Implications for the Conservation of Zabelia tyaihyonii Based on Genotyping-By-Sequencing.

Zabelia tyaihyonii (Nakai) Hisauti and H. Hara is a perennial shrub endemic to Republic of Korea that grows naturally in only a very limited region of the dolomite areas of Gangwon-do and Chungcheongbuk-do Provinces in the Republic of Korea. Given its geographical characteristics, it is more vulnerable than more widely distributed species. Despite the need for comprehensive information to support conservation, population genetic information for this species is very scarce. In this study, we analyzed the genetic diversity and population structure of 94 individuals from six populations of Z. tyaihyonii using a genotyping-by-sequencing (GBS) approach to provide important information for proper conservation and management. Our results, based on 3088 single nucleotide polymorphisms (SNPs), showed a mean expected heterozygosity (He) of 0.233, no sign of within-population inbreeding (GIS that was close to or even below zero in all populations), and a high level of genetic differentiation (FST = 0.170). Analysis of molecular variance (AMOVA) indicated that the principal molecular variance existed within populations (84.5%) rather than among populations (17.0%). We suggested that six management units were proposed for conservation considering Bayesian structure analysis and phylogenetic analysis, and given the various current situations faced by Z. tyaihyonii, it is believed that not only the in situ conservation but also the ex situ conservation should be considered.

Characterization and phylogenetic analysis of the complete chloroplast genome sequence of Pertya multiflora (Asteraceae), a rare and endangered wild plant species in China.

Pertya multiflora (Asteraceae) is a rare wild plant species narrowly distributed in Zhejiang province, China. In our present study, we assembled its complete chloroplast genome using high-throughput sequencing data. The results indicated that the whole chloroplast genome of P. multiflora was 153,396 bp in length. Its large single copy, small single copy, and inverted region sequences were 84,575 bp, 18,451 bp, and 25,185 bp. The P. multiflora chloroplast genome was composed of 134 genes, including 87 protein-coding genes, 37 tRNA genes, eight rRNA genes, and two pseudogenes. Phylogenetic analysis results showed that P. multiflora was grouped with Gerbera jamesonii, with 100% bootstrap support.

Non-Deep Simple Morphophysiological Dormancy and Germination Characteristics of Gentiana triflora var. japonica (Kusn.) H. Hara (Gentianaceae), a Rare Perennial Herb in Korea.

This study investigated the kind of seed dormancy and seed germination of Gentiana triflora var. japonica (Kusn.) H. Hara for developing a seed propagation method. The seeds were collected in October 2020 from plants at Mt. Sobaeksan, Korea. In a water imbibition experiment, seed weights increased by >101.9% of their initial masses over 12 h. Effects of incubation temperature (5, 15, 20, 25, 15/6, or 25/15 °C), cold stratification period (5 °C; 0, 4, 8, or 12 weeks), and gibberellic acid (GA3; 0, 10, 100, or 1000 mg∙L-1) and potassium nitrate treatment (KNO3; 0, 1000, 2000, or 4000 mg∙L-1) on seed germination were investigated to characterize seed dormancy. These seeds exhibited underdeveloped embryos during seed dispersal. The seeds failed to reach the final germination of 15.0% after treatment at 5, 15, 20, 25, 15/6, or 25/15 °C. After cold stratification for 8 weeks, the germination increased dramatically by >90.0% compared to that at 0 weeks. After the GA3 treatment, the germination reached >80.0% within 5 days. The final germination was 90.0% in the 100 mg∙L-1 GA3 treatment group. However, the KNO3 treatment had no effect on seed germination. Therefore, the G. triflora var. japonica seeds exhibited non-deep simple morphophysiological dormancy.

Characterization of the complete plastome sequence of Torreya grandis var. jiulongshanensis (Taxaceae), a rare and endangered plant species endemic to Zhejiang province, China.

Torreya grandis var. jiulongshanensis (Taxaceae) is an evergreen tree endemic to Zhejiang province, China, and it is listed as a rare plant species with extremely small populations. In our present study, we assembled the chloroplast (CP) genome of T. grandis var. jiulongshanensis using high-throughput sequencing data generated by an Illumina Hiseq X Ten platform. The results indicate that the complete CP genome is 136,705 bp in size with an overall guanine and cytosine content of 35.5%. The plastome has lost its quadripartite structure. In the CP genome, there are a total of 119 genes, including 82 protein-coding genes, 4 ribosomal RNA genes, and 33 transfer RNA genes. Phylogenetic analysis revealed that T. grandis var. jiulongshanensis is a sister species to T. grandis.

Characterization and phylogenetic analysis of the complete chloroplast genome sequence of Disanthus cercidifolius subsp. longipes (Hamamelidaceae), a rare and endangered wild plant species in China.

Disanthus cercidifolius subsp. longipes is a rare and endangered plant species. In our study, the complete chloroplast genome was assembled by using high-throughput DNA sequencing data. The whole CP genome is 158,076 bp in length, comprising of a large single-copy region of 87,148 bp, a small single-copy region of 18,300 bp, and two inverted repeat regions of 26,314 bp each. There are 136 genes in the genome, including 86 protein-coding genes, 40 transfer RNA genes, eight ribosomal RNA genes, and two pseudogenes (ndhK and ycf1). Phylogenetic results demonstrated that D. cercidifolius subsp. longipes grouped with other Hamamelidaceae species, with a support rate of 100%.

Habitat suitability and herbivores determine reintroduction success of an endangered legume.

Reintroductions of rare plants require detailed knowledge of habitat requirements, species interactions, and restoration techniques. Thus, incremental experimentation over many years may be required to develop adequate knowledge and techniques for successful reintroduction. To determine drivers of extinction in historical reintroductions of a federally endangered perennial (Astragalus bibullatus), we developed a reintroduction experiment to disentangle the relative importance of habitat quality, herbivores, and restoration technique on reintroduction success. In a factorial design, we manipulated access to vertebrate herbivores across different habitat types (mesic ecotone vs. xeric barren), and used founder populations comprised of more transplants and genetic sources than previous reintroduction attempts. In mesic ecotones where historical reintroductions failed, excluding herbivores, thinning woody encroachment to improve habitat quality, outplanting across a greater array of microhabitats, and increasing founder population size did not improve demographic rates over previous attempts. Compared to mesic ecotones, transplant survival rates and cumulative fruit production were more than two and ten times greater, respectively, in a xeric barren ecotone characterized by open, grassy, and dry microenvironmental conditions. Across all sites, herbivores decreased probabilities of survival and flowering of larger adult plants. Flowering rates were 80% greater inside relative to outside herbivore exclusion cages. Over a four-year period, only a single uncaged plant produced fruit. Our study demonstrates that habitat quality and vertebrate herbivory are key drivers of long-term persistence in rare plant reintroductions. Using incremental experiments that build on previous knowledge gained from long-term monitoring can improve reintroduction outcomes.

Lipophilic Metabolites and Anatomical Acclimatization of Cleome amblyocarpa in the Drought and Extra-Water Areas of the Arid Desert of UAE.

Plants adapt to different environmental conditions by developing structural and metabolic mechanisms. In this study, anatomical features and lipophilic metabolites were investigated in Cleome amblyocarpa Barr. & Murb., Cleomaceae plants growing in the arid desert of United Arab Emirates (UAE) in either low-water or extra-water areas, which were caused by the surrounding road run-off. The plant showed the presence of shaggy-like trichomes. The plant also developed special mechanisms to ensure its survival via release of lipophilic metabolites. The lipophilic metabolites, stained red with Sudan III, were apparently released by glandular trichomes and idioblasts of the shoot and roots, respectively. The identified lipophilic metabolites included those required for drought tolerance, protection against pathogens invasion, and detoxification. Plants growing in the low-water area caused an increase in the production of lipophilic metabolites-in particular, hydrocarbons and terpenoids. The lipophilic metabolites are known to provide the plant with unique waxy surfaces that reduce water loss and avoid penetration by pathogens. The release of lipid metabolites and the presence of shaggy-like trichomes represented unique features of the species that have never been reported. The provided chemical ecology information can be extended for several plant-related applications, particularly including drought tolerance.

Integrating the Rabinowitz rarity framework with a National Plant Inventory in South Korea.

Increasingly large presence-only survey datasets are becoming available for use in conservation assessments. Potentially, these records could be used to determine spatial patterns of plant species rarity and endemism. We test the integration of a large South Korean species record database with Rabinowitz rarity classes. Rabinowitz proposed seven classes of species rarity using three variables: geographic range, habitat specificity, and local population size. We estimated the range size and local abundance of 2,215 plant species from species occurrence records and habitat specificity as the number of landcover types each species' records were found in. We classified each species into a rarity class or as common, compared species composition by class to national lists, and mapped the spatial pattern of species richness for each rarity class. Species were classed to narrow or wide geographic ranges using 315 km, the average from a range size index of all species (D max), based on maximum distance between observations. There were four classes each within the narrow and wide range groups, sorted using cutoffs of local abundance and habitat specificity. Nationally listed endangered species only appeared in the narrow-range classes, while nationally listed endemic species appeared in almost all classes. Species richness in most rarity classes was high in northeastern South Korea especially for species with narrow ranges. Policy implications. Large presence-only surveys may be able to estimate some classes of rarity better than others, but modification to include estimates of local abundance and habitat types, could greatly increase their utility. Application of the Rabinowitz rarity framework to such surveys can extend their utility beyond species distribution models and can identify areas that need further surveys and for conservation priority. Future studies should be aware of the subjectivity of the rarity classification and that regional scale implementations of the framework may differ.

The complete chloroplast genome sequence of Berchemia berchemiifolia (Rhamnaceae).

The complete chloroplast genome sequence of Berchemia berchemiifolia, rare plant to Korea, was determined in this study. The total genome size was 160,410 bp in length, containing a pair of inverted repeats (IRs) of 26,514 bp, which were separated by a large single copy (LSC) and small single copy (SSC) of 88,627 bp and 18,755 bp, respectively. The overall GC contents of the chloroplast genome were 37.2%. One hundred twenty nine genes were annotated, including 84 protein coding genes, 37 tRNA genes, and 8 rRNA genes. In these genes, 18 genes contained one or two introns. The phylogenetic tree showed that Berchemia berchemiifolia was most closely related to Berchemiella wilsonii.

Effects of removing woody cover on long-term population dynamics of a rare annual plant (Agalinis auriculata): A study comparing remnant prairie and oldfield habitats.

Worldwide, grasslands are becoming shrublands/forests. In North America, eastern red cedar (Juniperus virginiana) often colonizes prairies. Habitat management can focus on woody removal, but we often lack long-term data on whether removal leads to population recovery of herbaceous plants without seeding. We undertook a long-term study (17 years) of numbers of the rare annual plant Agalinis auriculata in a gridwork of 100 m2 plots in adjacent prairie and oldfield sites in Kansas, USA. We collected data before and after removal of Juniperus virginiana at the prairie. Plant population sizes were highly variable at both sites and over time. High numbers of plants in a plot 1 year were often followed by low numbers the following year, suggesting negative density-dependence. Plant numbers were lowest with extensive woody cover and with low precipitation. After woody plant removal, A. auriculata increased dramatically in abundance and occupancy in most years; increases were also seen at the oldfield, suggesting later survey years were overall more favorable. Synthesis and applications: Removal of woody plants led to increased numbers of a rare annual prairie plant, without seeding. Multiple years of data were essential for interpretation given extreme temporal variability in numbers. The largest prairie population was 7 years following tree removal, showing that positive effects of management can last this long. This species also fared well in oldfield habitat, suggesting restoration opportunities. Given that land managers are busy, time-efficient field methods and data analysis approaches such as ours offer advantages. In addition to general linear models, we suggest Rank Occupancy-Abundance Profiles (ROAPs), a simple-to-use data visualization and analysis method. Creation of ROAPs for sites before and after habitat management helps reveal the degree to which plant populations are responding to management with changes in local density, changes in occupancy, or both.

Adaptive introductions: How multiple experiments and comparisons to wild populations provide insights into requirements for long-term introduction success of an endangered shrub.

Recovery of an imperiled plant species may require augmentation of existing populations or creation of new ones. Hundreds of such projects have been conducted over the last few decades, but there is a bias in the literature favoring successes over failures. In this paper, we evaluate a series of introductions that experimentally manipulated microhabitat and fire in an adaptive introduction framework. Between 2002 and 2012, we (and our collaborators) carried out ten introductions and augmentations of Florida ziziphus Pseudoziziphus (Condalia, Ziziphus) celata, a clonal shrub limited to very small populations and narrowly endemic to pyrogenic central Florida sandhills. Six of the introductions were designed as experiments to test hypotheses about how demographic performance was affected by microhabitat, fire, and propagule type. Introduced transplants had high survival (<90% annually), inconsistent growth, and little transition to reproduction, while introduced seeds had low germination and survival. Transplants were more efficient than seeds as translocation propagules. Shaded (vs. open) sites supported generally higher transplant and seedling survival and seed germination percentages, but growth responses varied among experiments. Supplemental irrigation increased transplant survival and seed germination, but otherwise seedling and plant survival and growth were not significantly affected. Contrary to expectations based on wild populations, introduced propagules have not been more successful in unshaded sites, suggesting that Florida ziziphus has broader microhabitat preferences than hypothesized. Compared to wild plants, introduced plants had similar survival and responses to fire, slower growth, and more delayed flowering. Introduced plants had no clonal spread. While no introduced population has demonstrated a capacity for long-term viability, one augmented population has flowered and produced viable fruits. Given that Florida ziziphus genets are long-lived, low levels of sexual reproduction may be adequate for the establishment of viable populations. Thus, after many translocations over more than a decade, it is premature to characterize any single translocation as a success or a failure, underscoring the need for a long view of translocation success.

Combined effects of heat shock, smoke and darkness on germination of Epacris stuartii Stapf., an endangered fire-prone Australian shrub.

Epacris stuartii (Epacridaceae) is an endangered heathland shrub in which seedling recruitment occurs almost exclusively after fire. Seed viability and multiple seed dormancy mechanisms were examined to explore why levels of seedling recruitment were low after some fires, despite high rates of pre-fire seed production. The individual and combined effects of heat shock, smoke derivatives and darkness on germination were tested in the laboratory using an orthogonal logit-linear model, a form of analysis ideally suited to hypotheses concerning multiple germination cues. Seed viability (56%) was found not to be limiting. After 89 days of incubation, germination was significantly enhanced by heat shock, smoke treatment and continuous darkness but there were no significant interactions. These effects were equal in magnitude and additive such that maximum germination (42% of viable seeds) was stimulated when all three treatments were combined, significantly less germination occurred in response to any two treatments combined (22-23%), any single treatment stimulated germination at levels (10-11%) significantly less than two treatments and the lowest levels of germination occurred among untreated seeds (4%). Relative to the untreated control, germination was stimulated by smoke derivatives in high concentrations but not in low concentrations. The effect of darkness diminished with time so that after 270 days of incubation darkness had no significant effect on germination, while heat and smoke still enhanced germination significantly, additively and interchangeably. More prolonged germination of seeds exposed to light on the soil surface than buried seed may spread the risk of desiccation of emerging seedlings over multiple rainfall events. The role of heat and smoke derivates suggests that fire management is a crucial tool for influencing seedling recruitment and hence the survival of E. stuartii at its only known location.