Karrikins are a novel class of naturally occurring plant growth regulators that promote seed germ... more Karrikins are a novel class of naturally occurring plant growth regulators that promote seed germination among a diverse range of species. Currently little is known about the mechanism by which these compounds overcome seed dormancy and initiate germination. The preparation of various karrikinolide derivatives provides an opportunity to investigate the karrikinolide mode of action at the cellular and molecular level. The first synthesis and biological evaluation of analogues suitable for use in metabolic labelling, affinity chromatography, photoaffinity labelling and NanoSIMS experiments are reported.
The smoke-derived compound karrikinolide (KAR(1)) shows significant potential as a trigger for th... more The smoke-derived compound karrikinolide (KAR(1)) shows significant potential as a trigger for the synchronous germination of seeds in a variety of plant-management contexts, from weed seeds in paddocks, to native seeds when restoring degraded lands. Understanding how KAR(1) interacts with seed physiology is a necessary precursor to the development of the compound as an efficient and effective management tool. This study tested the ability of KAR(1) to stimulate germination of seeds of the global agronomic weed Brassica tournefortii, at different hydration states, to gain insight into how the timing of KAR(1) applications in the field should be managed relative to rain events. Seeds of B. tournefortii were brought to five different hydration states [equilibrated at 15 % relative humidity (RH), 47 % RH, 96 % RH, fully imbibed, or re-dried to 15 % RH following maximum imbibition] then exposed to 1 nm or 1 microm KAR(1) for one of five durations (3 min, 1 h, 24 h, 14 d or no exposure)....
Cyanide is well known for its toxicity towards living organisms. Many plants use cyanide as a def... more Cyanide is well known for its toxicity towards living organisms. Many plants use cyanide as a defensive agent against herbivores, releasing it through the enzymatic hydrolysis of endogenous cyanogenic compounds. At low concentrations, cyanide has been proposed to have a regulatory role in many plant processes including stimulation of seed germination. However, no ecological role for cyanide in seed germination has been established. In the present study, we show that burning plant material produces the cyanohydrin, glyceronitrile. We also show that, in the presence of water, glyceronitrile is slowly hydrolysed to release cyanide that stimulates seed germination of a diverse range of fire-responsive species from different continents. We propose that glyceronitrile serves as an ecological store for cyanide and is an important cue for stimulating seed germination and landscape regeneration after fires.
Rock pools are small, geologically stable freshwater ecosystems that are both hydrologically and ... more Rock pools are small, geologically stable freshwater ecosystems that are both hydrologically and biologically isolated. They harbour high levels of plant endemism and experience environmental unpredictability driven by the presence of water over variable temporal scales. This study examined the hypothesis that the sediment seed bank in monsoon tropical freshwater rock pools would persist through one or more periods of desiccation, with seed dormancy regulating germination timing in response to rock pool inundation and drying events. Seeds were collected from seven dominant rock pool species, and germination biology and seed dormancy were assessed under laboratory conditions in response to light, temperature and germination stimulators (gibberellic acid, karrikinolide and ethylene). Field surveys of seedling emergence from freshwater rock pools in the Kimberley region of Western Australia were undertaken, and sediment samples were collected from 41 vegetated rock pools. Seedling emer...
Dispersal of plant propagules by ocean currents can result in long-distance dispersal and is impo... more Dispersal of plant propagules by ocean currents can result in long-distance dispersal and is important for the persistence of coastal species. However, the ability of such species to disperse via the ocean is often unknown because there is relatively little evidence that demonstrates that seeds or fruits can float and survive for extended periods in seawater. Furthermore, the seed or fruit traits, and intraspecific variation in these traits, that facilitate buoyancy remain largely unidentified. The genus Scaevola (L.) contains several widespread coastal species that may be capable of oceanic dispersal, such as S. crassifolia (Labill). We collected fruits of S. crassifolia along 700 km of a latitudinal environmental gradient. These fruits were used to determine the influence of fruit morphology and anatomy on fruit buoyancy. Morphological and anatomical variation in S. crassifolia was associated with dispersal potential. Our empirical data demonstrated that fruits with larger aeriferous mesocarp layers have greater buoyancy and, therefore, enhanced capacity for long range oceanic dispersal. Of three characters hypothesised to affect buoyancy (aeriferous mesocarp, air pockets in empty locules, and number of vascular cavities), only the properties of the mesocarp were significant. Intraspecific variation can significantly affect dispersal potential, and should not be overlooked in dispersal ecology.
Strigolactone hormones are derived from carotenoids via carlactone, and act through the α/β-hydro... more Strigolactone hormones are derived from carotenoids via carlactone, and act through the α/β-hydrolase D14 and the F-box protein D3/MAX2 to repress plant shoot branching. While MAX2 is also necessary for normal seedling development, D14 and the known strigolactone biosynthesis genes are not, raising the question of whether endogenous, canonical strigolactones derived from carlactone have a role in seedling morphogenesis. Here, we report the chemical synthesis of the strigolactone precursor carlactone, and show that it represses Arabidopsis shoot branching and influences leaf morphogenesis via a mechanism that is dependent on the cytochrome P450 MAX1. In contrast, both physiologically active Z-carlactone and the non-physiological E isomer exhibit similar weak activity in seedlings, and predominantly signal through D14 rather than its paralogue KAI2, in a MAX2-dependent but MAX1-independent manner. KAI2 is essential for seedling morphogenesis, and hence this early-stage development employs carlactone-independent morphogens for which karrikins from wildfire smoke are specific surrogates. While the commonly employed synthetic strigolactone GR24 acts non-specifically through both D14 and KAI2, carlactone is a specific effector of strigolactone signalling that acts through MAX1 and D14.
Several ecologically important plant families in Mediterranean biomes have seeds with morphophysi... more Several ecologically important plant families in Mediterranean biomes have seeds with morphophysiological dormancy (MPD) but have been poorly studied. The aim of this study was to understand the seed ecology of these species by focusing on the prominent, yet intractably dormant Australian genus Hibbertia. It was hypothesized that the slow germination in species of this genus is caused by a requirement for embryo growth inside the seed before germination, and that initiation of embryo growth is reliant upon a complex sequence of environmental cues including seasonal fluctuations in temperature and moisture, and an interplay with light and smoke. Using the results, the classification of the MPD level in species of Hibbertia is considered. Four species of Hibbertia in winter rainfall south-western Australia were selected. These species, whilst differing in geographic distributions, are variously sympatric, and all are important understorey components of plant communities. The following...
Aims Many terrestrial orchids have an obligate requirement for mycorrhizal associations to provid... more Aims Many terrestrial orchids have an obligate requirement for mycorrhizal associations to provide nutritional support from germination to establishment. This study will investigate the ability of orchid mycorrhizal fungi (OMF) to utilize a variety of nutrient sources in the nutrient-impoverished (low organic) soils of the Southwest Australian Floristic Region (SWAFR) in order to effectively compete, survive and sustain the orchid host. † Methods Mycorrhizal fungi representing key OMF genera were isolated from three common and widespread species: Pterostylis recurva, Caladenia flava and Diuris corymbosa, and one rare and restricted species: Drakaea elastica. The accessibility of specific nutrients was assessed by comparing growth including dry biomass of OMF in vitro on basal CN MMN liquid media. † Key Results Each of the OMF accessed and effectively utilized a wide variety of nutrient compounds, including carbon (C) sources, inorganic and organic nitrogen (N) and inorganic and organic phosphorus (P). The nutrient compounds utilized varied between the genera of OMF, most notably sources of N. † Conclusions These results suggest that OMF can differentiate between niches (micro-niche specialization) in a constrained, highly resource-limited environment such as the SWAFR. Phosphorus is the most limited macronutrient in SWAFR soils and the ability to access phytate by OMF indicates a characterizing functional capacity of OMF from the SWAFR. Furthermore, compared with OMF isolated from the rare D. elastica, OMF associating with the common P. recurva produced far greater biomass over a wider variety of nutritional sources. This suggests a broader tolerance for habitat variation providing more opportunities for the common orchid for recruitment and establishment at a site.
The efficacy of several sugars and polyalcohols in preculture medium was investigated using Anigo... more The efficacy of several sugars and polyalcohols in preculture medium was investigated using Anigozanthos 6iridis ssp terraspectans Hopper (Haemodoraceae), a threatened plant species endemic to the south west of Western Australia. A vitrification protocol involving preculturing of shoot apices for 3 days on different concentrations of sugars and polyalcohols, followed by incubation in plant vitrification solution 2 (PVS2) for 25 min, prior to immersion in liquid nitrogen (LN) and warming resulted in shoot tip survival ranging from 34 to 84%. High levels of survival were obtained with polyalcohols, compared to sucrose, glucose, trehalose and raffinose when used at the same molarity (0.4 M) or at the equivalent concentration of total hydroxyl (OH) groups present in molecules. In both cases glycerol proved more effective. When polyalcohols (ribitol and erythritol) with similar stereochemical arrangement of OH groups as glycerol were examined, at the same molarity (0.4 M) and with equivalent OH numbers, higher survival was achieved when the total number of OH groups present was the same as glycerol. Additionally, when the structural isomers mannitol/sorbitol and ribitol/xylitol were compared at the same molarity (0.4 M), the isomer with the higher number of OH groups along the same side resulted in significantly higher levels of post-LN survival. We propose that the mode of action of polyalcohols is based not on molarity, but on the total number of OH groups present in the medium. Furthermore, based on these results we propose that the orientation of OH groups is a determining factor in effective cryopreservation.
This study investigated the effects of 3-methyl-2Hfuro[2,3-c ]pyran-2-one, a germination active b... more This study investigated the effects of 3-methyl-2Hfuro[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.
Karrikins are a novel class of naturally occurring plant growth regulators that promote seed germ... more Karrikins are a novel class of naturally occurring plant growth regulators that promote seed germination among a diverse range of species. Currently little is known about the mechanism by which these compounds overcome seed dormancy and initiate germination. The preparation of various karrikinolide derivatives provides an opportunity to investigate the karrikinolide mode of action at the cellular and molecular level. The first synthesis and biological evaluation of analogues suitable for use in metabolic labelling, affinity chromatography, photoaffinity labelling and NanoSIMS experiments are reported.
The smoke-derived compound karrikinolide (KAR(1)) shows significant potential as a trigger for th... more The smoke-derived compound karrikinolide (KAR(1)) shows significant potential as a trigger for the synchronous germination of seeds in a variety of plant-management contexts, from weed seeds in paddocks, to native seeds when restoring degraded lands. Understanding how KAR(1) interacts with seed physiology is a necessary precursor to the development of the compound as an efficient and effective management tool. This study tested the ability of KAR(1) to stimulate germination of seeds of the global agronomic weed Brassica tournefortii, at different hydration states, to gain insight into how the timing of KAR(1) applications in the field should be managed relative to rain events. Seeds of B. tournefortii were brought to five different hydration states [equilibrated at 15 % relative humidity (RH), 47 % RH, 96 % RH, fully imbibed, or re-dried to 15 % RH following maximum imbibition] then exposed to 1 nm or 1 microm KAR(1) for one of five durations (3 min, 1 h, 24 h, 14 d or no exposure)....
Cyanide is well known for its toxicity towards living organisms. Many plants use cyanide as a def... more Cyanide is well known for its toxicity towards living organisms. Many plants use cyanide as a defensive agent against herbivores, releasing it through the enzymatic hydrolysis of endogenous cyanogenic compounds. At low concentrations, cyanide has been proposed to have a regulatory role in many plant processes including stimulation of seed germination. However, no ecological role for cyanide in seed germination has been established. In the present study, we show that burning plant material produces the cyanohydrin, glyceronitrile. We also show that, in the presence of water, glyceronitrile is slowly hydrolysed to release cyanide that stimulates seed germination of a diverse range of fire-responsive species from different continents. We propose that glyceronitrile serves as an ecological store for cyanide and is an important cue for stimulating seed germination and landscape regeneration after fires.
Rock pools are small, geologically stable freshwater ecosystems that are both hydrologically and ... more Rock pools are small, geologically stable freshwater ecosystems that are both hydrologically and biologically isolated. They harbour high levels of plant endemism and experience environmental unpredictability driven by the presence of water over variable temporal scales. This study examined the hypothesis that the sediment seed bank in monsoon tropical freshwater rock pools would persist through one or more periods of desiccation, with seed dormancy regulating germination timing in response to rock pool inundation and drying events. Seeds were collected from seven dominant rock pool species, and germination biology and seed dormancy were assessed under laboratory conditions in response to light, temperature and germination stimulators (gibberellic acid, karrikinolide and ethylene). Field surveys of seedling emergence from freshwater rock pools in the Kimberley region of Western Australia were undertaken, and sediment samples were collected from 41 vegetated rock pools. Seedling emer...
Dispersal of plant propagules by ocean currents can result in long-distance dispersal and is impo... more Dispersal of plant propagules by ocean currents can result in long-distance dispersal and is important for the persistence of coastal species. However, the ability of such species to disperse via the ocean is often unknown because there is relatively little evidence that demonstrates that seeds or fruits can float and survive for extended periods in seawater. Furthermore, the seed or fruit traits, and intraspecific variation in these traits, that facilitate buoyancy remain largely unidentified. The genus Scaevola (L.) contains several widespread coastal species that may be capable of oceanic dispersal, such as S. crassifolia (Labill). We collected fruits of S. crassifolia along 700 km of a latitudinal environmental gradient. These fruits were used to determine the influence of fruit morphology and anatomy on fruit buoyancy. Morphological and anatomical variation in S. crassifolia was associated with dispersal potential. Our empirical data demonstrated that fruits with larger aeriferous mesocarp layers have greater buoyancy and, therefore, enhanced capacity for long range oceanic dispersal. Of three characters hypothesised to affect buoyancy (aeriferous mesocarp, air pockets in empty locules, and number of vascular cavities), only the properties of the mesocarp were significant. Intraspecific variation can significantly affect dispersal potential, and should not be overlooked in dispersal ecology.
Strigolactone hormones are derived from carotenoids via carlactone, and act through the α/β-hydro... more Strigolactone hormones are derived from carotenoids via carlactone, and act through the α/β-hydrolase D14 and the F-box protein D3/MAX2 to repress plant shoot branching. While MAX2 is also necessary for normal seedling development, D14 and the known strigolactone biosynthesis genes are not, raising the question of whether endogenous, canonical strigolactones derived from carlactone have a role in seedling morphogenesis. Here, we report the chemical synthesis of the strigolactone precursor carlactone, and show that it represses Arabidopsis shoot branching and influences leaf morphogenesis via a mechanism that is dependent on the cytochrome P450 MAX1. In contrast, both physiologically active Z-carlactone and the non-physiological E isomer exhibit similar weak activity in seedlings, and predominantly signal through D14 rather than its paralogue KAI2, in a MAX2-dependent but MAX1-independent manner. KAI2 is essential for seedling morphogenesis, and hence this early-stage development employs carlactone-independent morphogens for which karrikins from wildfire smoke are specific surrogates. While the commonly employed synthetic strigolactone GR24 acts non-specifically through both D14 and KAI2, carlactone is a specific effector of strigolactone signalling that acts through MAX1 and D14.
Several ecologically important plant families in Mediterranean biomes have seeds with morphophysi... more Several ecologically important plant families in Mediterranean biomes have seeds with morphophysiological dormancy (MPD) but have been poorly studied. The aim of this study was to understand the seed ecology of these species by focusing on the prominent, yet intractably dormant Australian genus Hibbertia. It was hypothesized that the slow germination in species of this genus is caused by a requirement for embryo growth inside the seed before germination, and that initiation of embryo growth is reliant upon a complex sequence of environmental cues including seasonal fluctuations in temperature and moisture, and an interplay with light and smoke. Using the results, the classification of the MPD level in species of Hibbertia is considered. Four species of Hibbertia in winter rainfall south-western Australia were selected. These species, whilst differing in geographic distributions, are variously sympatric, and all are important understorey components of plant communities. The following...
Aims Many terrestrial orchids have an obligate requirement for mycorrhizal associations to provid... more Aims Many terrestrial orchids have an obligate requirement for mycorrhizal associations to provide nutritional support from germination to establishment. This study will investigate the ability of orchid mycorrhizal fungi (OMF) to utilize a variety of nutrient sources in the nutrient-impoverished (low organic) soils of the Southwest Australian Floristic Region (SWAFR) in order to effectively compete, survive and sustain the orchid host. † Methods Mycorrhizal fungi representing key OMF genera were isolated from three common and widespread species: Pterostylis recurva, Caladenia flava and Diuris corymbosa, and one rare and restricted species: Drakaea elastica. The accessibility of specific nutrients was assessed by comparing growth including dry biomass of OMF in vitro on basal CN MMN liquid media. † Key Results Each of the OMF accessed and effectively utilized a wide variety of nutrient compounds, including carbon (C) sources, inorganic and organic nitrogen (N) and inorganic and organic phosphorus (P). The nutrient compounds utilized varied between the genera of OMF, most notably sources of N. † Conclusions These results suggest that OMF can differentiate between niches (micro-niche specialization) in a constrained, highly resource-limited environment such as the SWAFR. Phosphorus is the most limited macronutrient in SWAFR soils and the ability to access phytate by OMF indicates a characterizing functional capacity of OMF from the SWAFR. Furthermore, compared with OMF isolated from the rare D. elastica, OMF associating with the common P. recurva produced far greater biomass over a wider variety of nutritional sources. This suggests a broader tolerance for habitat variation providing more opportunities for the common orchid for recruitment and establishment at a site.
The efficacy of several sugars and polyalcohols in preculture medium was investigated using Anigo... more The efficacy of several sugars and polyalcohols in preculture medium was investigated using Anigozanthos 6iridis ssp terraspectans Hopper (Haemodoraceae), a threatened plant species endemic to the south west of Western Australia. A vitrification protocol involving preculturing of shoot apices for 3 days on different concentrations of sugars and polyalcohols, followed by incubation in plant vitrification solution 2 (PVS2) for 25 min, prior to immersion in liquid nitrogen (LN) and warming resulted in shoot tip survival ranging from 34 to 84%. High levels of survival were obtained with polyalcohols, compared to sucrose, glucose, trehalose and raffinose when used at the same molarity (0.4 M) or at the equivalent concentration of total hydroxyl (OH) groups present in molecules. In both cases glycerol proved more effective. When polyalcohols (ribitol and erythritol) with similar stereochemical arrangement of OH groups as glycerol were examined, at the same molarity (0.4 M) and with equivalent OH numbers, higher survival was achieved when the total number of OH groups present was the same as glycerol. Additionally, when the structural isomers mannitol/sorbitol and ribitol/xylitol were compared at the same molarity (0.4 M), the isomer with the higher number of OH groups along the same side resulted in significantly higher levels of post-LN survival. We propose that the mode of action of polyalcohols is based not on molarity, but on the total number of OH groups present in the medium. Furthermore, based on these results we propose that the orientation of OH groups is a determining factor in effective cryopreservation.
This study investigated the effects of 3-methyl-2Hfuro[2,3-c ]pyran-2-one, a germination active b... more This study investigated the effects of 3-methyl-2Hfuro[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.
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