Australian Tree Research Articles

Valorisation of Three Underutilised Native Australian Plants: Phenolic and Organic Acid Profiles and In Vitro Antimicrobial Activity.

Tasmannia lanceolata, Diploglottis bracteata and Syzygium aqueum are understudied native Australian plants. This study aimed to characterise the non-anthocyanin phenolic and organic acid profiles of the aqueous extracts obtained from the leaves of T. lanceolata and fruits of D. bracteata and S. aqueum by UHPLC-Q-Orbitrap-MS/MS and UHPLC-TQ-MS/MS. A total of 39, 22, and 27 non-anthocyanin polyphenols were tentatively identified in T. lanceolata, D. bracteata, and S. aqueum extracts, respectively. Furthermore, sugars and ascorbic acid contents as well as in vitro antioxidant and antimicrobial activities of the extracts were determined. Response surface methodology was applied to achieve an extract blend with a strong inhibitory effect against Pseudomonas viridiflava, the main cause of soft rot in vegetables, Bacillus subtilis, Rhodotorula diobovata and Alternaria alternata. The identified compounds including organic acids (e.g., quinic, citric and malic acids) and polyphenols (e.g., catechin, procyanidins, and ellagitannins) might contribute to the observed antimicrobial activity. Furthermore, this study provides the most comprehensive phenolic profiles of these three underutilised native Australian plants to date.

The membrane activity of the antimicrobial peptide caerin 1.1 is pH dependent.

Histidine-rich antimicrobial peptides (AMPs) are an important class of membrane-active peptides that can provide a scaffold to induce pH-sensitive interactions with cell membranes. The Australian tree frog AMP, caerin 1.1 (Cae-1), has three histidine residues, which modify the interactions with neutral and negatively charged lipid systems. NMR and MD simulations showed that in an acidic environment in comparison to physiological conditions, Cae-1 induced greater perturbation of the lipid dynamics and water penetration into the membrane interior. Lipid packing, chemical environment, and dynamics of the phospholipid headgroup were monitored using 31P solid-state NMR. In particular, the CODEX technique revealed that Cae-1 has a large impact on the slow re-orientation of the lipids. 2H solid-state NMR showed that Cae-1 ordered the acyl chains across the hydrophobic core. Interestingly, titration of the paramagnetic Mn2+ into the lipid-AMP systems resulted in extensive quenching of the 13C solid-state NMR signals of the lipid headgroup and acyl chain carbons. These results supported the MD results that showed Cae-1 was mainly inserted within the lipid bilayer structure of both neutral and negatively charged membranes, with the charged residues dragging the water and phosphate groups inwards. This could be an early step in the mechanism of membrane disruption by histidine-rich AMPs and indicated that Cae-1 is potentially forming toroidal pores in bilayers of neutral and negatively charged lipid membranes, especially under acidic conditions.

The membrane activity of the antimicrobial peptide caerin 1.1 is pH dependent

Antimicrobial peptides are an important class of membrane-active peptides that can provide alternatives or complements to classic antibiotics. Among the many classes of AMPs, the histidine-rich family is of particular interest since they may induce pH-sensitive interactions with cell membranes. The AMP caerin 1.1 (Cae-1), from Australian tree frogs, has three histidine residues, and thus we studied the pH dependence of its interactions with model cell membranes. Using NMR spectroscopy and molecular dynamics simulations, we showed that Cae-1 induced greater perturbation of the lipid dynamics and water penetrations within the membrane interior in an acidic environment compared with physiological conditions. Using 31P solid-state NMR, the packing, chemical environment, and dynamics of the lipid headgroup were monitored. 2H solid-state NMR showed that Cae-1 ordered the acyl chains of the hydrophobic core of the bilayer. These results supported the molecular dynamics data, which showed that Cae-1 was mainly inserted within the lipid bilayer for both neutral and negatively charged membranes, with the charged residues pulling the water and phosphate groups inward. This could be an early step in the mechanism of membrane disruption by histidine-rich antimicrobial peptides and indicated that Cae-1 acts via a transmembrane mechanism in bilayers of neutral and anionic phospholipid membranes, especially in acidic conditions.

Temperature, nutrient availability, and species traits interact to shape elevation responses of Australian tropical trees

Elevation gradients provide natural laboratories for investigating tropical tree ecophysiology in the context of climate warming. Previously observed trends with increasing elevation include decreasing stem diameter growth rates (GR), increasing leaf mass per area (LMA), higher root-to-shoot ratios (R:S), increasing leaf δ13C, and decreasing leaf δ15N. These patterns could be driven by decreases in temperature, lower soil nutrient availability, changes in species composition, or a combination thereof. We investigated whether these patterns hold within the genus Flindersia (Rutaceae) along an elevation gradient (0–1,600 m) in the Australian Wet Tropics. Flindersia species are relatively abundant and are important contributors to biomass in these forests. Next, we conducted a glasshouse experiment to better understand the effects of temperature, soil nutrient availability, and species on growth, biomass allocation, and leaf isotopic composition. In the field, GR and δ15N decreased, whereas LMA and δ13C increased with elevation, consistent with observations on other continents. Soil C:N ratio also increased and soil δ15N decreased with increasing elevation, consistent with decreasing nutrient availability. In the glasshouse, relative growth rates (RGR) of the two lowland Flindersia species responded more strongly to temperature than did those of the two upland species. Interestingly, leaf δ13C displayed an opposite relationship with temperature in the glasshouse compared with that observed in the field, indicating the importance of covarying drivers in the field. Leaf δ15N increased in nutrient-rich compared to nutrient-poor soil in the glasshouse, like the trend in the field. There was a significant interaction for δ15N between temperature and species; upland species showed a steeper increase in leaf δ15N with temperature than lowland species. This could indicate more flexibility in nitrogen acquisition in lowland compared to upland species with warming. The distinguishing feature of a mountaintop restricted Flindersia species in the glasshouse was a very high R:S ratio in nutrient-poor soil at low temperatures, conditions approximating the mountaintop environment. Our results suggest that species traits interact with temperature and nutrient availability to drive observed elevation patterns. Capturing this complexity in models will be challenging but is important for making realistic predictions of tropical tree responses to global warming.

Small Leaves, Big Diversity: Citizen Science and Taxonomic Revision Triples Species Number in the Carnivorous Drosera microphylla Complex (D. Section Ergaleium, Droseraceae).

The carnivorous Drosera microphylla complex from southwest Western Australia comprises a group of rare, narrowly endemic species that are potentially threatened by habitat destruction and illegal collection, thus highlighting a need for accurate taxonomic classification to facilitate conservation efforts. Following extensive fieldwork over two decades, detailed studies of both Australian and European herbaria and consideration of both crucial contributions by citizen scientists and social media observations, nine species of the D. microphylla complex are here described and illustrated, including four new species: D. atrata, D. hortiorum, D. koikyennuruff, and D. reflexa. The identities of the previously described infraspecific taxa D. calycina var. minor and D. microphylla var. macropetala are clarified. Both are here lectotypified, reinstated, and elevated to species rank. A replacement name, D. rubricalyx, is provided for the former taxon. Key morphological characters distinguishing the species of this complex include the presence or absence of axillary leaves, lamina shape, petal colour, filament shape, and style length. A detailed identification key, comparison figures, and a distribution map are provided. Six of the nine species are recommended for inclusion on the Priority Flora List under the Conservation Codes for Western Australian Flora and Fauna.

Integrating seed microbiome knowledge into restoration and ex situ conservation of native Australian plants

Context Seeds harbour a diversity of microbes, which in some plants aid with germination and establishment. Seeds form a critical part in the lifecycle of plants and a role in many conservation and restoration activities. Aims Because this is an emerging field in seed biology, we aim to highlight the key research gaps of interest to seed on the basis of restoration and ex situ conservation. Methods We identify knowledge gaps associated with the seed endophytic microbiome of native Australian plants through undertaking a literature review. Additionally, culturing methods were used to identify the fungal seed endophytes of five native Australian species. Key results We identified a diversity of taxa within the native seed and show three taxa that are common to all study hosts. Sampling seed from additional hosts at a site and additional sites of a host species showed new fungal diversity. Our literature review showed that little information is available on native seed microbiomes and we identified four key areas where research gaps exist, linking with seed-based restoration practices. Conclusions We provide evidence that there is a complex and diverse seed microbiome within some Australian native plants and suggest ways that it could be integrated into restoration and conservation practices. Implications We propose that by taking into consideration the presence of a seed microbiome and its potential impacts on plant health, seed microbiomes could be used as one method to restore microbial diversity into an ecosystem and to contribute to the seedling microbiome and plant health at restored sites.

Genomic characterization supporting the development of new food and crop options from the Australian flora

Genome analysis is facilitating the use of crop wild relatives in the Australian flora for food crop improvement. Options for domestication of new food species from this diverse flora are also being revealed.

The Potential of Yellow Gum (Eucalyptus leucoxylon F. Muell.) as an Urban Street Tree: An Assessment of Species Performance in the City of Greater Melbourne, Australia

"Background: In contrast to trees from northern hemisphere genera, there has been little research into the selection and vegetative propagation of Australian native tree species for use as street trees. Eucalyptus leucoxylon F. Muell. is one of a few eucalypts occurring in south-eastern Australia with bright coloured fl owers and is highly regarded as an ornamental tree that fl owers readily. It is propagated from seed, but progeny typically show seedling variability and diversity. Eucalyptus leucoxylon was identifi ed as the most widely planted eucalypt in the streets of the city of greater Melbourne, Australia. Methods: This research assessed 300 E. leucoxylon street trees growing across the city of greater Melbourne for their performance against arboricultural criteria relating to canopy structure and density, straightness of the trunk, health (assessed on canopy, trunk, and branch condition, production of exudates, and presence of fungal fruiting bodies), fl ower colour, and root systems. Results: The results showed that E. leucoxylon was a suitable street tree species with most specimens showing good habit, vigour, and health. Discussion: The trees had traits such as live crown ratio, height, fl ower colour, and capacity to cope with pruning that are considered appropriate for a street tree. Their dense canopies and high live crown ratios provide shade that can reduce the urban heat island (UHI) effect. Conclusion: This suggests the species has the potential to be a successful street tree not only in Australia, but in other parts of the world where it has been grown successfully in forestry plantations.

Rodents may be important pollinators of banksias in subtropical Australia

Knowledge of the role of non-flying mammals in the pollination of the Australian flora is still incomplete. We captured non-flying mammals visiting Banksia ericifolia in north-east New South Wales and sampled pollen from their heads. The grassland melomys (Melomys burtoni) was captured >4 times more often than the house mouse (Mus musculus) or the swamp rat (Rattus lutreolus). Nocturnal pollen loads (mean 295 ± 53 grains) measured on the melomys are among the highest for an Australian non-flying mammal. Pollen removal from flowers at night was widespread. These observations suggest the grassland melomys is likely to play an important role in the pollination of subtropical B. ericifolia.

Characterizing Effects of Microbial Biostimulants and Whole-Soil Inoculums for Native Plant Revegetation

Soil microbes play important roles in plant health and ecosystem functioning, however, they can often be disturbed or depleted in degraded lands. During seed-based revegetation of such sites there is often very low germination and seedling establishment success, with recruitment of beneficial microbes to the rhizosphere one potential contributor to this problem. Here we investigated whether Australian native plant species may benefit from planting seed encapsulated within extruded seed pellets amended with one of two microbe-rich products: a commercial vermicast extract biostimulant or a whole-soil inoculum from a healthy reference site of native vegetation. Two manipulative glasshouse trials assessing the performance of two Australian native plant species (Acacia parramattensis and Indigofera australis) were carried out in both unmodified field-collected soil (trial 1) and in the same soil reduced in nutrients and microbes (trial 2). Seedling emergence and growth were compared between pelleted and bare-seeded controls and analyzed alongside soil nutrient concentrations and culturable microbial community assessments. The addition of microbial amendments maintained, but did not improve upon, high levels of emergence in both plant species relative to unamended pellets. In trial 1, mean time to emergence of Acacia parramattensis seedlings was slightly shorter in both amended pellet types relative to the standard pellets, and in trial 2, whole-soil inoculum pellets showed significantly improved growth metrics. This work shows that there is potential for microbial amendments to positively affect native plant emergence and growth, however exact effects are dependent on the type of amendment, the plant species, and the characteristics of the planting site soil.

Embedded in the Bark: Kimberley Boab Trees as Sites of Historical Archaeology

This paper discusses the Australian boab tree and its potential for research as living historical archaeology. Boab trees play an important role in the economy, culture, and cosmology of Indigenous people in northwest Australia and continue to hold a powerful presence in the Kimberley region today. Working with Nyikina and Mangala Traditional Owners we have undertaken to document examples of this iconic tree and its cultural and historical associations, particularly in the form of carvings and inscriptions embedded in the bark. Focusing on four individual trees located in the Kimberley region of northwest Australia, we propose that the modification of boab trees, as a practice undertaken by both Indigenous and non-Indigenous people, offers important insights into the everyday lives and historic events that shaped this cultural landscape.

Cold pretreatment for cryopreservation of an endangered Australian native plant species Gossia fragrantissima

Cold pretreatment for cryopreservation of an endangered Australian native plant species Gossia fragrantissima

Detection of eucalyptus oil adulteration in Australian tea tree oil using UV–Vis and fluorescence spectroscopy

Pure Australian tea tree oil is a high-value commodity; however, cheaper tea tree oils are often adulterated with other substances such as eucalyptus oil. Detection of adulteration typically requires time-consuming analysis with expensive equipment. Consequently, rapid methods of screening for adulteration would be beneficial for the industry. This study examined whether the rapid, non-destructive and low-cost methods of ultraviolet-visible and fluorescence spectroscopy could be used to detect adulteration of tea tree oil with eucalyptus oil. Ultraviolet-visible spectroscopy showed moderate accuracy for predicting adulterant concentration (root mean square error of cross-validation (RMSECV) of 6.8% v/v). Moving window analysis was used to reduce the model wavelength range to just 55 nm, without a loss in predictive accuracy. Fluorescence spectroscopy also performed well, with an RMSECV of 2.9% v/v. Consequently, these techniques may be suitable for rapidly and cheaply screening tea tree oil samples for gross adulteration.

Genetic variation in fire recovery and other fire-related traits in a global eucalypt species

To understand the potential of forests to adapt to wildfire, we studied the genetic architecture of fire-related structural, damage and recovery traits in a globally important Australian forest tree species, Eucalyptus globulus. Fourteen traits were evaluated in an outcrossed F2 population in a field trial in Tasmania, Australia, which was burnt by a wildfire 14 years after planting. The trial also included open-pollinated families of the grandparental dwarf and tall ecotypes used to produce the F2 population. We studied the phenotypic correlations within the F2 population and performed quantitative trait loci (QTL) analyses using a linkage map comprised of 472 markers. Ecotype comparisons revealed that almost all traits were under genetic control, with trees of the dwarf ecotype significantly more damaged and mainly recovering from lignotubers, whereas tall ecotype trees mainly recovered from epicormic resprouts extending for a variable height up the stem. Within the F2, tree size was negatively correlated with fire damage and positively correlated with recovery. Genetic control of fire-related traits was confirmed by the detection of 38 QTL in the F2 population. These QTL accounted for 4 to 43% of the phenotypic variation in these traits. Several QTL co-located and likely reflect pleiotropic effects. However, many independent QTL were detected, including QTL for crown consumption and trunk scorch, epicormic resprouting, resprout herbivory, and seedling establishment. The QTL detected argue that many genetically controlled mechanisms are responsible for variation in fire damage and recovery.

International Laws and Regulations on Cutting of Trees

Forests are a gift of nature. Association of trees is with human beings from time immemorial. Trees knowno boundaries. Principles to save trees are universal in nature. Laws on protection of trees evolved with theevolution of laws. It is a modern concern to save trees as the increase in a world population, the demand forexploitation of trees for timber and furniture has increased. The United Nations as world organization hastaken keen interest in the protection of trees. In the present article I have dealt with the laws,rules andregulation for protection of trees in Australia, Brazil, Canada and the United States of America.

Bioassay Guided Fractionation Protocol for Determining Novel Active Compounds in Selected Australian Flora.

A large variety of unique and distinct flora of Australia have developed exceptional survival methods and phytochemicals and hence may provide a significant avenue for new drug discovery. This study proposes a bioassay guided fractionation protocol that maybe robust and efficient in screening plants with potential bioactive properties and isolating lead novel compounds. Hence, five native Australian plants were selected for this screening process, namely Pittosporum angustifolium (Gumbi gumbi), Terminalia ferdinandiana (Kakadu plum, seeds (KPS), and flesh (KPF)), Cupaniopsis anacardioides (Tuckeroo, seeds (TKS) and flesh (TKF)), Podocarpus elatus (Illawarra plum, seeds (IPS) and flesh (IPF)) and Pleiogynium timoriense (Burdekin plum, seeds (BPS) and flesh (BPF)). The methanolic extracts of the plants samples were analysed for Total phenolic content (TPC) and antioxidant capacity measure by FRAP. The highest values were found in the KPF which were 12,442 ± 1355 mg GAE/ 100 g TPC and 16,670 ± 2275 mg TXE/100 g antioxidant capacity. Extracts of GGL was deemed to be most potent with complete cell inhibition in HeLa and HT29, and about 95% inhibition in HuH7 cells. Comparative activity was also seen for KPS extract, where more than 80% cell inhibition occurred in all tested cell lines. Dose-dependent studies showed higher SI values (0.72-1.02) in KPS extracts than GGL (0.5-0.73). Microbial assays of the crude extracts were also performed against five bacterial strains commonly associated with causing food poisoning diseases were selected (Gram positive-Staphylococcus aureus and Gram negative-Escherichia coli, Salmonella typhi and Pseudomonas aeruginosa bacteria). KPF extracts were effective in suppressing microbial growth of all tested bacterial strains except for P. aeruginosa, while TKS and TKF were only slightly effective against S. aureus. Due to the potential of the GGL crude extract to completely inhibit the cells compared to KPS, it was further fractionated and tested against the cell lines. HPLC phenolic profiling of the crude extracts were performed, and numerous peak overlaps were evident in the fruit extracts. The KPF extracts demonstrated the strongest peaks which was coherent with the fact that it had the highest TPC and antioxidant capacity values. A high occurrence of t-ferulic acid in the GGL extracts was found which may explain the cytotoxic activity of GGL extracts. Peaks in KPS and KPF extracts were tentatively identified as gallic acid, protocatechuic acid, 4-hydroxybenzoic acid and syringic acid and possibly ellagic acid. HPLC time-based fractionation of the GGL extract (F1-F5) was performed and Dose dependent cytotoxic effects were determined. It was construed that F1, having the highest SI value for HeLa, HT29 and HuH7 (1.60, 1.41 and 1.67, respectively) would be promising for further fractionation and isolation process.

LC-MS/MS Characterization of Phenolic Metabolites and Their Antioxidant Activities from Australian Native Plants.

Polyphenols are considered vital bioactive compounds beneficial for human health. The Australian flora is enriched with polyphenols which are not fully characterized yet. Thus, the main objective of this study was to identify and characterize the Australian native sandalwood nuts, wattle seeds, lemongrass, and old man saltbush for phenolic compounds and their antioxidant activities. In this study, we tentatively identified a total of 155 phenolic compounds including 25 phenolic acids, 55 flavonoids, 22 isoflavonoids, 22 tannins, 22 lignans, 33 stilbenes, 33 coumarins and derivatives, 12 tyrosols and derivatives, and 6 phenolic terpenes. The highest total phenolic content (TPC) (15.09 ± 0.88 mg GAE/g) was quantified in lemongrass, while the lowest TPC (4.17 ± 0.33 mg GAE/g) was measured in wattle seeds. The highest total flavonoid content (TFC) and total condensed tannins (TCT) were measured in lemongrass and wattle seeds, respectively. A total of 18 phenolic metabolites were quantified/semi-quantified in this experiment. Lemongrass contains a vast number of phenolic metabolites.

A genome resource for Acacia, Australia's largest plant genus.

Acacia (Leguminosae, Caesalpinioideae, mimosoid clade) is the largest and most widespread genus of plants in the Australian flora, occupying and dominating a diverse range of environments, with an equally diverse range of forms. For a genus of its size and importance, Acacia currently has surprisingly few genomic resources. Acacia pycnantha, the golden wattle, is a woody shrub or tree occurring in south-eastern Australia and is the country's floral emblem. To assemble a genome for A. pycnantha, we generated long-read sequences using Oxford Nanopore Technology, 10x Genomics Chromium linked reads, and short-read Illumina sequences, and produced an assembly spanning 814 Mb, with a scaffold N50 of 2.8 Mb, and 98.3% of complete Embryophyta BUSCOs. Genome annotation predicted 47,624 protein-coding genes, with 62.3% of the genome predicted to comprise transposable elements. Evolutionary analyses indicated a shared genome duplication event in the Caesalpinioideae, and conflict in the relationships between Cercis (subfamily Cercidoideae) and subfamilies Caesalpinioideae and Papilionoideae (pea-flowered legumes). Comparative genomics identified a suite of expanded and contracted gene families in A. pycnantha, and these were annotated with both GO terms and KEGG functional categories. One expanded gene family of particular interest is involved in flowering time and may be associated with the characteristic synchronous flowering of Acacia. This genome assembly and annotation will be a valuable resource for all studies involving Acacia, including the evolution, conservation, breeding, invasiveness, and physiology of the genus, and for comparative studies of legumes.

Post‐fire restoration of Sphagnum bogs in the Tasmanian Wilderness World Heritage Area, Australia

Sphagnum bogs in Australia are small, with a limited distribution, but of high conservation value. They are restricted to cool, wet environments that are typically fire free and are poorly adapted to recover from fire disturbance, unlike most Australian flora. Increased fire activity due to anthropogenic climate change is threatening Sphagnum bogs. This increased threat has stimulated interest in their restoration. Compared with the northern hemisphere, there have been few studies of the ecology of Sphagnum restoration in the southern hemisphere. Here, we report on a field experiment in Tasmania, in an area burned by an extensive fire in 2016. We investigated the role of shade, fertilizer and transplants, factors demonstrated to be important in the restoration of burnt bogs on the Australian mainland. Treatments commenced three to 4 years after the fire. Overall, we found that fire‐damaged Sphagnum recovers very slowly, and that there was no recovery in severely burned areas. The addition of shade increased recovery of damaged Sphagnum, but fertilizer was harmful, even to healthy Sphagnum. Transplants in fire‐killed Sphagnum grew poorly in both moderately and severely burnt Sphagnum areas. Our findings support the use of shading in post‐fire Sphagnum recovery projects, although further work is required to determine the optimal approach and duration of providing shade.

Grevillea tesselata Olde (Proteaceae: Grevilleoideae: Hakeinae), a rare new species of uncertain affinity from the Avon Wheatbelt region of south-west Western Australia.

Grevillea tesselata Olde is described here as a new species from south-west Western Australia, known only from a small population in a fragmented roadside landscape, and previously recognised under the phrase name Grevillea sp. Trayning (W. Johnston WJ 071). Following the Flora of Australia, the new species keys to the Grevillea Acacioides Group which comprises only three species, G. endlicheriana Meisn., G. acacioides C.A.Gardner ex McGill. and G. gordoniana C.A.Gardner. A binary assessment of 50 morphological characters presented here supports the view that G. acacioides and G. endlicheriana are sister species. A key to the new species is provided and its distribution updated. Grevillea tesselata has a Priority One Conservation Code according to the Western Australian Herbarium.