Smoke Enhanced Seed Germination for Mine Rehabilitation in the Southwest of Western Australia

Cool smoke treatments were applied to unmined Eucalyptus marginata (jarrah) forest soils, rehabilitated bauxite mine soils, and broadcast seed to determine if enhancement in germination could be effected with a view toward maximizng the establishment of species in bauxite mines in Western Australia. Forest sites showed a 48-fold increase in total germinants from the soil seed bank when treated with aerosol smoke. Newly returned bauxite mine soils showed a greater than threefold increase in total germinants after the same treatment. There were also significant increases in the number of species germinating in response to the aerosol smoke treatment in both the forest and the mined soils. Similarly, application of smoked water to the soil seed bank in previously mined sites elicited a significant positive germination response, increasing total germinants and species numbers by 56 and 33%, respectively. Treatment of mixed seed lots with aerosol smoke before broadcast resulted in highly significant improvement in germination when compared to untreated seed. Both total number of germinants, and number of species emerging from mined sites were positively influenced (85% and 34% increases, respectively). Ten target species were used to determine the relative effectiveness of different methods of smoke treatment on the germination of broadcast seed. Nine of the species involved displayed a promotive effect with at least two of the treatments. Generally, however, aerosol smoking of seed before broadcast proved to be the more effective approach. As a result of these findings, all broadcast seed for use in Alcoa's bauxite mined areas in the southwest of Western Australia is now routinely smoke treated before application.     

Seed germination traits of desert perennials

While understanding that seed germination is crucial for ecological restoration activities, the seed traits of desert perennials are understudied. We experimentally determined germination traits of 43 species from 14 families from Hummock grasslands in the Great Sandy Desert, Australia. We defined morphological and physiological seed traits of framework species required for restoration and investigated the effects of fire and temperature on seed germination. We classified dormancy and explored the effect of Karrikinolide, a fire cue derived from smoke, on germination. Seeds of 38 (88%) out of 43 species were dormant: 13 (30%) with physical and 25 (58%) with physiological dormancy. Karrikinolide promoted seed germination of 9 (21%) species across all life-forms except trees, and widened the range of germination temperatures and increased germination rate of one species. Although high germination percentages were obtained over a wide temperature range, germination rate was affected by temperature. Non-dormant seeds and seeds pre-treated to overcome physical dormancy germinated quickly, with times to 50% germination of 1–5 days. Dormancy class differed between life-forms and families. Fast germination of non-dormant seeds is a trait that allows seeds to germinate during short periods of moisture availability. An absence of under-developed embryos is consistent with the global trends for hot deserts. A response to Karrikinolide shows that seed germination is related to a fire cue. These results will inform land managers of effective seed pre-treatments prior to seed broadcasting for restoration, and information on seed germination temperatures and rates will improve the understanding of when and where seeds could germinate in restored sites.     

Restoring the Victorian Western (Basalt) Plains grassland. 1. Laboratory trials of viability and germination, and the implications for direct seeding

Summary   The Victorian Western (Basalt) Plains grassland is one of Australia's most threatened plant communities. Practitioners using seed for its restoration need to know whether seed can be sown fresh or whether it requires an after-ripening period. This study assessed the viability and germination of freshly harvested wild seed from 64 grassland species indigenous to the Basalt Plains of western Victoria. The seed was collected as part of a broader experiment that examined the potential of direct-sown complex seed mixes for the restoration of grassland communities. The germination of fresh seed at 25°C varied widely between species. Comparisons with tetrazolium viability tests for each species indicated varying levels of dormancy within the species pool. Germination separated into three broad responses at day 28. One-third of the species failed to germinate, one-third germinated at 1% to 50% and the remaining species germinated between 51% and 100%. Therefore, if the aim of a sowing was the rapid and synchronous establishment of most of the sown species, the use of fresh seed in restoration could be problematic. After 3 months of dry storage, eight species were re-tested for germination. Each of the selected species had shown high viability but low initial germination. Only two species significantly increased their total germination at 25°C. The annual species, Triptilodiscus pygmaeus , increased its total germination from 6% as fresh seed to 99% after dry storage. Testing the viability and germination capacity of freshly harvested seed from a large and diverse sample of native grassland species demonstrated that many of the species were unlikely to germinate rapidly or synchronously when sown in complex seed mixes soon after harvest. This finding has implications for the scheduling and management of restoration projects that rely on the use of such seed.     

Dormancy and germination: How does the crop seed decide?

Whether seeds germinate or maintain dormancy is decided upon through very intricate physiological processes. Correct timing of these processes is most important for the plants life cycle. If moist conditions are encountered, a low dormancy level causes pre‐harvest sprouting in various crop species, such as wheat, corn and rice, this decreases crop yield and negatively impacts downstream industrial processing. In contrast, a deep level of seed dormancy prevents normal germination even under favourable conditions, resulting in a low emergence rate during agricultural production. Therefore, an optimal seed dormancy level is valuable for modern mechanised agricultural systems. Over the past several years, numerous studies have demonstrated that diverse endogenous and environmental factors regulate the balance between dormancy and germination, such as light, temperature, water status and bacteria in soil, and phytohormones such as ABA (abscisic acid) and GA (gibberellic acid). In this updated review, we highlight recent advances regarding the molecular mechanisms underlying regulation of seed dormancy and germination processes, including the external environmental and internal hormonal cues, and primarily focusing on the staple crop species. Furthermore, future challenges and research directions for developing a full understanding of crop seed dormancy and germination are also discussed.     

Factors controlling seed germination of the Iberian critically endangered Pseudomisopates (Antirrhinaceae)

Many seeds are dormant when shed from the mother plant. This unique characteristic of plants poses challenges in conservation and many different treatments have traditionally been used to break dormancy. When germination only occurs under certain circumstances, recruitment may be insufficient and the viability of plant populations may be threatened. A marked dormancy was previously identified in the Iberian critically endangered species Pseudomisopates rivas-martinezii. The present study aimed to determine the magnitude of dormancy by estimating seed viability under different germination treatments: heat, ash, stratification, gibberellin addition, and darkness. Our results indicate that there were significant differences in viability across plant populations and treatments. Maximum seed germination was obtained under ash addition, although cold stratification alone gave a considerable enhancement. However, gibberellic acid did not improve germination and darkness had a diminishing effect. In conclusion, a cold period is required for the species to germinate, coupled with the effects of fire: (1) ash, (2) vegetation clearings, and (3) resprouting, which are major factors triggering seed germination. Although these conditions are widely found in the field, we hypothesize that a more dramatic situation may have tackled this species before the increase of human-mediated fires in the past millennia.     

光信号与激素调控种子休眠和萌发研究进展

休眠是种子植物在长期进化过程中产生的适应性性状, 通过抑制种子在不适宜的环境中萌发进而保证植物能够在逆境中生存。此外, 休眠有助于种子的长距离运输和扩散, 因此休眠对种子延续和物种保存具有重要意义。种子由休眠向萌发的发育转变不仅关系到物种的繁衍, 而且对保证农业生产中作物的产量和品质也具有重要作用。种子的休眠和萌发受到内源激素和外源光信号的共同调控。其中, 外源光信号主要通过调控内源ABA和GA的生物合成及信号转导进而调控种子休眠和萌发。该文系统综述了外源光信号和内源激素调控种子休眠和萌发的作用通路以及两类信号通路之间的交互作用, 旨在为农业生产中利用光和激素调控种子休眠与萌发提供参考。     

光信号与激素调控种子休眠和萌发研究进展

休眠是种子植物在长期进化过程中产生的适应性性状, 通过抑制种子在不适宜的环境中萌发进而保证植物能够在逆境中生存。此外, 休眠有助于种子的长距离运输和扩散, 因此休眠对种子延续和物种保存具有重要意义。种子由休眠向萌发的发育转变不仅关系到物种的繁衍, 而且对保证农业生产中作物的产量和品质也具有重要作用。种子的休眠和萌发受到内源激素和外源光信号的共同调控。其中, 外源光信号主要通过调控内源ABA和GA的生物合成及信号转导进而调控种子休眠和萌发。该文系统综述了外源光信号和内源激素调控种子休眠和萌发的作用通路以及两类信号通路之间的交互作用, 旨在为农业生产中利用光和激素调控种子休眠与萌发提供参考。     

Bauxite Mining Restoration by Alcoa World Alumina Australia in Western Australia: Social,Political, Historical,and Environmental Contexts

Alcoa World Alumina Australia mines bauxite under lease agreements with the Government of Western Australia. The leases lie in the Darling Range to the east of Perth, the capital and major population center. In addition to bauxite and other mineral ores, the Darling Range is a major potable water source and harbors a species-rich forest dominated by Jarrah (Eucalyptus marginata), a significant commercial timber. Conservation and recreation are important land uses in the region. Social and political pressures have led to stringent governmental requirements for restoration. In addition, a summer drought period, a soil deficient in most nutrients, water management challenges, an introduced disease, caused by Phytophthora cinnamomi Rands, and a post-mining ecosystem that must be conducive to the prescribed burning management of the region pose significant challenges to successful restoration. Alcoa presently mines and restores approximately 550 ha per annum. Although the “footprint” at the end of the life of the mining operations represents only about 4% of the total forest estate, Alcoa is committed to restoring the forest values of the region of all lands impacted by mining. The major objective of restoration is to enhance or maintain forest values by restoring habitat and structural characteristics of the native forest environment. Completion criteria for Alcoa’s mine restoration have been developed. The original Alcoa mine at Jarrahdale has been rehabilitated, and in 2005, a 975-ha area received a “certificate of completion” and was returned to the management control of the State of Western Australia.     

Morphological side effects of using gibberellic acid to induce germination: consequences for the study of seed dormancy

To assess the evolutionary significance of persistent seed banks, phenotypes of naturally germinating seeds must be compared with those that remain dormant under the same environmental conditions. Dormant seeds can often be induced to germinate by application of gibberellic acid (GA). However, this method is valid only if there are no phenotypic “side effects” of GA that could confound comparisons between dormant and naturally germinating seeds. We examined this assumption in Lesquerella fendleri, a short-lived perennial mustard of the desert Southwest. We exposed 3840 seeds from 16 maternal sibships to two different GA treatments (0 or 1 g/L GA) in two different germination environments (greenhouse and growth chamber), and measured germination and postgermination traits. As expected, application of GA increased germination. GA also had strong and long-lasting effects on seedling morphology. Seeds that received GA developed into seedlings that were taller, with fewer but longer leaves, than seeds that did not receive GA. Effects of GA on both dormancy and postgermination traits varied among maternal sibships. Our results indicate that for this species and this concentration of GA, morphological effects can be substantial. Further study is required to determine whether such side effects are found for lower concentrations of GA, or under conditions that encourage faster seedling growth. Nonetheless, the present results illustrate the importance of testing potential confounding effects of GA in studies of the evolution of seed dormancy and its influence on postgermination traits.     

Vegetation Succession After Bauxite Mining in Western Australia

Alcoa World Alumina Australia has been rehabilitating bauxite mines in the jarrah forest of Western Australia for more than 35 years. An experiment was established in 1988 using three different seed treatments (legume and small understorey mix, small understorey mix only, and no seed) and two fertilizer treatments (N and P, and P only). The objectives of this study were to (1) document vegetation changes in the first 14 years after bauxite mining; (2) assess whether the vegetation is becoming more similar to the unmined forest; and (3) gain a better understanding of successional processes. Seed treatments significantly affected 13 of the 14 measured vegetation characteristics. Native species richness was higher in seeded than in unseeded sites at 1, 2, and 5 years of age, whereas diversity and evenness were generally higher at all assessment ages. Exotic species density was higher in unseeded than in seeded sites from 5 years onward, whereas richness was higher from 8 years onward. Nitrogen fertilizer significantly increased exotic species richness, density, and cover. Ephemerals dominated plant density in all rehabilitation treatments over time, whereas seeder species dominated cover. In contrast, resprouting species dominated density and cover in the unmined forest. Orchids were the only species that were not present in the first year in rehabilitated sites but increased in abundance over time. Vegetation composition in rehabilitated areas did not become more similar to the unmined forest during the 14 years since seeding, instead strongly reflected the initial species mix. Rehabilitated bauxite mines appear to follow the initial floristic composition model of succession.     

羊草种子休眠机制及破除方法研究

羊草种子休眠程度深、发芽率低是限制栽培利用的重要因子.采用不同破除羊草种子休眠的方法,测定各处理对种子萌发的影响,以探索破除羊草种子休眠的有效途径.结果显示:(1)刺破种皮的裸种子较完整种子的萌发率、吸水速率、生活力分别由对照的6%、63%、0%显著增加到60%、86%、94%.(2)完整羊草种子分别用清水浸种1 d、30% NaOH浸种80 min、清水浸种1 d后用30% NaOH浸种60min其萌发率由6%分别显著提高到36%、60%、84%,而各浓度赤霉素处理完整种子其萌发率较对照均无显著变化. (3)采用清水浸种1 d后用30% NaOH处理60 min,再施加200 μg/g GA3综合处理,可使羊草完整种子的发芽率由6%提高到91%,接近其种子生活力94%.研究表明,羊草种子的稃与种皮不影响种子水分的吸收,但影响种子对GA3的吸收、不同程度地阻碍大分子物质的渗入、限制羊草种子内部萌发抑制物的渗出,从而引起种子休眠;分析认为稃和种皮以及种子内部萌发抑制物质是引起羊草种子休眠的主要原因.     

Germination behaviour of Astroloma xerophyllum (Ericaceae), a species with woody indehiscent endocarps

The dispersal unit of many Ericaceae comprises an ovoid drupe with a woody indehiscent endocarp, and diaspores of this type are notoriously difficult to germinate for most members of this widely distributed family. Within the biodiverse south‐west of Western Australia, more than 200 drupaceous species of Ericaceae have been described, more than 50 of which are considered to be rare and threatened, requiring significant conservation action in the near future. In this paper, we investigate the germination ecology of the common Australian endemic, Astroloma xerophyllum, as a proxy for closely related threatened taxa, focusing on the ex situ and in situ germination requirements of seeds and indehiscent endocarps. Each endocarp possessed up to seven locules and means of 2.0–3.4 seeds per endocarp from the two collections used in this study. Seeds were up to 2.74 mm in length and 100% viable. Embryos were linear, differentiated and approximately 1.3 mm in length. Seeds within endocarps imbibed water to 28%, whereas excised seeds became hydrated to 44%. Fifty‐five per cent of seeds extracted from endocarps germinated on water agar alone and 100% germinated when presoaked in gibberellic acid. Seeds remaining inside intact endocarps failed to germinate unless treated with a germination promoter and incubated for more than 20 weeks. Rapid germination of seeds in intact endocarps was promoted by soaking endocarps in gibberellic acid and incubating them in 100% O₂. Embryos grew substantially in length within seeds prior to germination, and thus seeds have morphophysiological dormancy. Seeds under natural conditions required several seasons to germinate to any degree. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 160 , 299–311.     

Effects of storage,mucilage presence,photoperiod, thermoperiod and salinity on germination of Farsetia aegyptia Turra (Brassicaceae) seeds: implications for restoration and seed banks in Arabian Desert

Seed viability and germination are key factors in the success of restoration efforts, especially when stored seeds are used. However, the effect of seed storage on germination of most of the native Arabian species is not well documented. We investigated the effect of storage time and role of the seed mucilage in regulating germination, dormancy, salinity tolerance and consequential survival strategy of F. aegyptia in an unpredictable arid desert setting. Effect of light and temperature during germination was studied under two photoperiods and two thermoperiods using intact and de-mucilaged seeds. Presence of mucilage and thermoperiod did not affect the germination. However, seed collection year and photoperiod had a highly significant effect on the germination. Increasing salinity levels decreased the germination of F. aegyptia but ungerminated seeds were able to germinate when salinity stress was alleviated. Seed storage at room temperature enhances the germination percentage, indicating that F. aegyptia seeds have physiological dormancy and it can be alleviated by after-ripening at dry storage. In addition, F. aegyptia seeds show ability to germinate at lower salinity concentration and remain viable even at higher saline conditions, indicating their adaptability to cope with such harsh environmental conditions.     

Dormancy-breaking and germination requirements for seeds of Symphoricarpos orbiculatus (Caprifoliaceae)

Fruits (drupes) of Symphoricarpos orbiculatus ripen in autumn and are dispersed from autumn to spring. Seeds (true seed plus fibrous endocarp) are dormant at maturity, and they have a small, linear embryo that is underdeveloped. In contrast to previous reports, the endocarp and seed coat of S. orbiculatus are permeable to water; thus, seeds do not have physical dormancy. No fresh seeds germinated during 2 wk of incubation over a 15°/6°-35°/20°C range of thermoperiods in light (14-h photoperiod); gibberellic acid and warm or cold stratification alone did not overcome dormancy. One hundred percent of the seeds incubated in a simulated summer → autumn → winter → spring sequence of temperature regimes germinated, whereas none of those subjected to a winter → spring sequence did so. That is, cold stratification is effective in breaking dormancy only after seeds first are exposed to a period of warm temperatures. Likewise, embryos grew at cold temperatures only after seeds were exposed to warm temperatures. Thus, the seeds of S. orbiculatus have nondeep complex morphophysiological dormancy. As a result of dispersal phenology and dormancy-breaking requirements, in nature most seeds that germinate do so the second spring following maturity; a low to moderate percentage of the seeds may germinate the third spring. Seeds can germinate to high percentages under Quercus leaf litter and while buried in soil; they have little or no potential to form a long-lived soil seed bank.     

Seed germination of GA-insensitive sleepy1 mutants does not require RGL2 protein disappearance in Arabidopsis

We explore the roles of gibberellin (GA) signaling genes SLEEPY1 (SLY1) and RGA-LIKE2 (RGL2) in regulation of seed germination in Arabidopsis thaliana, a plant in which the hormone GA is required for seed germination. Seed germination failure in the GA biosynthesis mutant ga1-3 is rescued by GA and by mutations in the DELLA gene RGL2, suggesting that RGL2 represses seed germination. RGL2 protein disappears before wild-type seed germination, consistent with the model that GA stimulates germination by causing the SCF(SLY1) E3 ubiquitin ligase complex to trigger ubiquitination and destruction of RGL2. Unlike ga1-3, the GA-insensitive sly1 mutants show variable seed dormancy. Seed lots with high seed dormancy after-ripened slowly, with stronger alleles requiring more time. We expected that if RGL2 negatively controls seed germination, sly1 mutant seeds that germinate well should accumulate lower RGL2 levels than those failing to germinate. Surprisingly, RGL2 accumulated at high levels even in after-ripened sly1 mutant seeds with 100% germination, suggesting that RGL2 disappearance is not a prerequisite for seed germination in the sly1 background. Without GA, several GA-induced genes show increased accumulation in sly1 seeds compared with ga1-3. It is possible that the RGL2 repressor of seed germination is inactivated by after-ripening of sly1 mutant seeds.     

Cracking the case: Seed traits and phylogeny predict time to germination in prairie restoration species

Traits are important for understanding how plant communities assemble and function, providing a common currency for studying ecological processes across species, locations, and habitat types. However, the majority of studies relating species traits to community assembly rely upon vegetative traits of mature plants. Seed traits, which are understudied relative to whole‐plant traits, are key to understanding assembly of plant communities. This is particularly true for restored communities, which are typically started de novo from seed, making seed germination a critical first step in community assembly and an early filter for plant establishment. We experimentally tested the effects of seed traits (mass, shape, and embryo to seed size ratio) and phylogeny on germination response in 32 species commonly used in prairie grassland restoration in the Midwestern USA, analyzing data using time‐to‐event (survival) analysis. As germination is also influenced by seed dormancy, and dormancy break treatments are commonly employed in restoration, we also tested the effects of two pretreatments (cold stratification and gibberellic acid application) on time to germination. Seed traits, phylogeny, and seed pretreatments all affected time to germination. Of all traits tested, variables related to seed shape (height and shape variance) best predicted germination response, with high‐variance (i.e., pointier and narrower) seeds germinating faster. Phylogenetic position (the location of species on the phylogenetic tree relative to other tested species) was also an important predictor of germination response, that is, closely related species showed similar patterns in time to germination. This was true despite the fact that all measured seed traits showed phylogenetic signal, therefore phylogeny provided residual information that was not already captured by measured seed traits. Seed traits, phylogenetic position, and germination pretreatments were important predictors of germination response for a suite of species commonly used in grassland restoration. Shape traits were especially important, while mass, often the only seed trait used in studies of community assembly, was not a strong predictor of germination timing. These findings illustrate the ecological importance of seed traits that are rarely incorporated into functional studies of plant communities. This information can also be used to advance restoration practice by guiding restoration planning and seed mix design.     

Is broad-scale smoke–water application always a useful tool for improving seedling emergence in post-mining restoration? Evidence from jarrah forest restoration in Western Australia

It has been widely advocated that smoke–water application to topsoil can substantially improve restoration success by enhancing seed germination. This is despite few studies having tested the effects of smoke–water on seedling emergence in field-scale restoration trials. Here we report the effects of applying a commercially available smoke solution (Regen 2000®), at rates between 0 and 100 mL m^− 2, on jarrah forest sites being restored after bauxite mining in the southwest of Western Australia. Smoke solutions stimulated the seed germination of a range of species in laboratory experiments. In addition, smoke–water stimulated germination of Stylidium affine seeds sown directly into the first field experiment. However, apart from the effect on sown S. affine seeds, smoke–water application had no effect on subsequent seedling numbers, species richness or the relative proportion of seedlings in different growth-form categories in either of the two field experiments. These findings suggest that smoke–water application does not always ensure enhanced restoration outcomes.     

Grazing by Kangaroos Limits the Establishment of the Grass Trees Xanthorrhoea gracilis and X. preissii in Restored Bauxite Mines in Eucalypt Forest of Southwestern Australia

This study examined factors affecting germination, survival, and growth of the grass trees Xanthorrhoea gracilis and X. preissii on newly rehabilitated bauxite mine pits in the jarrah forest of southwestern Australia. Grazing by kangaroos (Macropus fuliginosus) was the major factor in reducing survival and growth of both species during the first 2 years. Provision of artificial grazing protection increased survival and growth (plant mass) of both species by 3‐fold. Grazing by native vertebrates has not previously been identified as affecting mine restoration in Western Australia. Initial germination rates from sown seeds of X. preissii at eight replicate sites ranged from 25 to 64% with a mean of 42%. Corresponding figures for X. gracilis were 5–42% with a mean of 17%. Germination of X. gracilis was greater on heavier, moister soils, but X. preissii germinated better on sandier soils. High levels of initial germination did not ensure high survival. Plants of both species grew bigger and survived better on the lighter, sandy soils. Xanthorrhoea seedlings located in the depressions created by the ripping process grew larger than seedlings on the slopes of the riplines. The presence of plants of other species did not have a significant effect on survival. However, these plants facilitated the growth of both species when artificial grazing protection was unavailable. Plants of other species reduced the growth rates of Xanthorrhoea seedlings where artificial grazing protection was provided. Artificial shade by itself had no significant effect on growth of either species. In rehabilitated bauxite mines in the jarrah forest, the provision of grazing protection is recommended to ensure successful establishment and early survival of Xanthorrhoea spp.     

Germination response of Arabian desert species to gibberellic acid and potassium nitrate seed treatment

Abstract

Desert plant species commonly use seed dormancy to prevent germination during unfavorable environmental conditions and thus increase the probability of seedling survival. Seed dormancy presents a challenge for restoration ecology, particularly in desert species for which our knowledge of dormancy regulation is limited. In the present study the effect of gibberellic acid (GA₃) and potassium nitrate (KNO₃) on seed dormancy release was investigated on eight Arabian desert species. Both treatments significantly enhanced the germination of most species tested. GA₃ was more effective than KNO₃ in enhancing germination percentage, reducing mean germination time and synchronizing the germination in most of the studied species. Light requirement during germination was species-specific, but in general the presence of light promoted germination more effectively when combined with KNO₃ and GA₃. The wide variation in dormancy and germination requirements among the tested species is indicative of distinct germination niches, which might assist their co-existence in similar habitat/environmental conditions. Seed pre-treatments that optimize germination in this habitat must therefore be assessed for individual species to improve the outcomes of ecological restoration.     

Implications of Seedling Emergence to Site Restoration following Bauxite Mining in Western Australia

Seedling emergence of 12 selected northern jarrah (Eucalyptus marginata Donn ex Smith) forest species were investigated to assist Alcoa of Australia Ltd. in maximizing the establishment of topsoil species in rehabilitated bauxite mining sites. The species, which encompassed a range of seed weights (0.024 mg to 87 mg), plant families, seed-storage types, life forms, and germination requirements, were placed on the soil surface and at depths of 1, 2, 5, 10, and 15 cm under controlled conditions in a glasshouse. Ability to emerge from deep burial was found to depend on seed size for species that annually release their seed to the topsoil but not for species that store their seed on the plant. All selected species were capable of emerging from 2 cm depth of burial, but eight of the 12 species were either unable to emerge from 5 cm or showed a significant reduction in emergence from 5 cm depth of burial compared to optimally buried seed. This group included two small-seeded species, Stylidium calcaratum and Chamaescilla corymbosa; the major forest dominant, Eucalyptus marginata; the serotinous canopy-borne seed of Hakea amplexicaulis; and the wind-dispersed seed of Xanthorrhoea gracilis. A few seeds of the legume species Kennedia coccinea, Acacia pulchella, and Bossiaea aquifolium established seedlings from depths of 15 cm. Currently, Alcoa removes the upper 15 cm of topsoil separately from the underlying soil prior to the commencement of mining. This topsoil is respread at a similar depth following mining as part of the rehabilitation procedure. It is recommended that Alcoa continue to strip topsoil to a depth of 15 cm but investigate the option of re-spreading topsoil onto rehabilitated pits at a shallower depth to maximize establishment via the soil seed bank.