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Numerous studies have analysed the relationship between C4 plant cover and climate. However, few have examined how different C4 taxa vary in their response to climate, or how environmental factors alter C4:C3 abundance. Here we investigate (a) how proportional C4 plant cover and richness (relative to C3) responds to changes in climate and local environmental factors, and (b) if this response is consistent among families. Proportional cover and richness of C4 species were determined at 541 one-hectare plots across Australia for 14 families. C4 cover and richness of the most common and abundant families were regressed against climate and local parameters. C4 richness and cover in the monocot families Poaceae and Cyperaceae increased with latitude and were strongly positively correlated with January temperatures, however C4 Cyperaceae occupied a more restricted temperature range. Seasonal rainfall, soil pH, soil texture, and tree cover modified proportional C4 cover in both families. Eudicot families displayed considerable variation in C4 distribution patterns. Proportional C4 Euphorbiaceae richness and cover were negatively correlated with increased moisture availability (i.e. high rainfall and low aridity), indicating they were more common in dry environments. Proportional C4 Chenopodiaceae richness and cover were weakly correlated with climate and local environmental factors, including soil texture. However, the explanatory power of C4 Chenopodiaceae models were poor, suggesting none of the factors considered in this study strongly influenced Chenopodiaceae distribution. Proportional C4 richness and cover in Aizoaceae, Amaranthaceae, and Portulacaceae increased with latitude, suggesting C4 cover and richness in these families increased with temperature and summer rainfall, but sample size was insufficient for regression analysis. Results demonstrate the unique relationships between different C4 taxa and climate, and the significant modifying effects of environmental factors on C4 distribution. Our work also revealed C4 families will not exhibit similar responses to local perturbations or climate., Competing Interests: The authors have declared that no competing interests exist.

The photosynthetic pathway of plants is a fundamental trait that influences terrestrial environments from the local to global level. The distribution of different photosynthetic pathways in Australia is expected to undergo a substantial shift due to climate change and rising atmospheric CO 2 ; however, tracking change is hindered by a lack of data on the pathways of species, as well as their distribution and relative cover within plant communities. Here we present the photosynthetic pathways for 2428 species recorded across 541 plots surveyed by Australia's Terrestrial Ecosystem Research Network (TERN) between 2011 and 2017. This dataset was created to facilitate research exploring trends in vegetation change across Australia. Species were assigned a photosynthetic pathway using published literature and stable carbon isotope analysis of bulk tissue. The photosynthetic pathway of species can be extracted from the dataset individually, or used in conjunction with vegetation surveys to study the occurrence and abundance of pathways across the continent. This dataset will be updated as TERN's plot network expands and new information becomes available.

Leaf area (LA), mass per area (LMA), nitrogen per unit area (N area ) and the leaf-internal to ambient CO 2 ratio (χ) are fundamental traits for plant functional ecology and vegetation modelling. Here we aimed to assess how their variation, within and between species, tracks environmental gradients. Measurements were made on 705 species from 116 sites within a broad north-south transect from tropical to temperate Australia. Trait responses to environment were quantified using multiple regression; within- and between-species responses were compared using analysis of covariance and trait-gradient analysis. Leaf area, the leaf economics spectrum (indexed by LMA and N area ) and χ (from stable carbon isotope ratios) varied almost independently among species. Across sites, however, χ and LA increased with mean growing-season temperature (mGDD 0 ) and decreased with vapour pressure deficit (mVPD 0 ) and soil pH. LMA and N area showed the reverse pattern. Climate responses agreed with expectations based on optimality principles. Within-species variability contributed < 10% to geographical variation in LA but > 90% for χ, with LMA and N area intermediate. These findings support the hypothesis that acclimation within individuals, adaptation within species and selection among species combine to create predictable relationships between traits and environment. However, the contribution of acclimation/adaptation vs species selection differs among traits., (© 2020 The Authors. New Phytologist © 2020 New Phytologist Trust.)

Premise: Within closed-canopy forests, vertical gradients of light and atmospheric CO 2 drive variations in leaf carbon isotope ratios, leaf mass per area (LMA), and the micromorphology of leaf epidermal cells. Variations in traits observed in preserved or fossilized leaves could enable inferences of past forest canopy closure and leaf function and thereby habitat of individual taxa. However, as yet no calibration study has examined how isotopic, micro- and macromorphological traits, in combination, reflect position within a modern closed-canopy forest or how these could be applied to the fossil record., Methods: Leaves were sampled from throughout the vertical profile of the tropical forest canopy using the 48.5 m crane at the Daintree Rainforest Observatory, Queensland, Australia. Carbon isotope ratios, LMA, petiole metric (i.e., petiole-width 2 /leaf area, a proposed proxy for LMA that can be measured from fossil leaves), and leaf micromorphology (i.e., undulation index and cell area) were compared within species across a range of canopy positions, as quantified by leaf area index (LAI)., Results: Individually, cell area, δ 13 C, and petiole metric all correlated with both LAI and LMA, but the use of a combined model provided significantly greater predictive power., Conclusions: Using the observed relationships with leaf carbon isotope ratio and morphology to estimate the range of LAI in fossil floras can provide a measure of canopy closure in ancient forests. Similarly, estimates of LAI and LMA for individual taxa can provide comparative measures of light environment and growth strategy of fossil taxa from within a flora., (© 2020 Cheesman et al. American Journal of Botany published by Wiley Periodicals LLC on behalf of Botanical Society of America.)

Rationale: Purification of α-cellulose from plant tissues is commonly conducted to facilitate the reliable measurement of stable isotope ratios. Prior research has shown that different plant species and tissues react differently to standardised cellulose extraction techniques. Thus, no single method can be applied to all materials and careful consideration must be undertaken when selecting an extraction technique., Methods: In order to evaluate their suitability for use on Melaleuca quinquenervia leaves, a suite of eight different cellulose extraction techniques were tested. Leaves of this species are preserved in perched lakes on southeast Queensland's sand islands and are a focus of ongoing palaeoclimate research. Elemental analyser/isotope ratio mass spectrometry was used to measure stable carbon and oxygen isotopic ratios and sample composition was measured using Fourier transform infrared spectroscopy., Results: We demonstrate that the standard Brendel extraction technique, particularly with a higher reagent volume and longer boiling time, produces cellulose with the lowest spread in isotopic ratios among replicates, and with the fewest impurities detected by Fourier transform infrared spectroscopy. We also show that pre-treating the leaves to extract leaf wax lipids in order to enable paired analysis from the same sub-fossil leaves does not significantly affect the quality of the isotopic results., Conclusions: The standard Brendel method allows the most precise stable carbon and oxygen information to be retrieved from the leaves of M. quinquenervia. This unlocks the potential to study palaeoclimate proxy records from our study site and potentially throughout the natural range of the species across eastern Australia, Papua New Guinea and New Caledonia., (Copyright © 2018 John Wiley & Sons, Ltd.)

Intraspecific plant functional trait variation provides mechanistic insight into persistence and can infer population adaptive capacity. However, most studies explore intraspecific trait variation in systems where geographic and environmental distances co-vary. Such a design reduces the certainty of trait-environment associations, and it is imperative for studies that make trait-environment associations be conducted in systems where environmental distance varies independently of geographic distance. Here we explored trait variation in such a system, and aimed to: (i) quantify trait variation of parent and offspring generations, and associate this variation to parental environments; (ii) determine the traits which best explain population differences; (iii) compare parent and offspring trait-trait relationships. We characterized 15 plant functional traits in eight populations of a shrub with a maximum separation ca. 100 km. Populations differed markedly in aridity and elevation, and environmental distance varied independently of geographic distance. We measured traits in parent populations collected in the field, as well as their offspring reared in greenhouse conditions. Parent traits regularly associated with their environment. These associations were largely lost in the offspring generation, indicating considerable phenotypic plasticity. An ordination of parent traits showed clear structure with strong influence of leaf area, specific leaf area, stomatal traits, isotope δ 13 C and δ 15 N ratios, and N area , whereas the offspring ordination was less structured. Parent trait-trait correlations were in line with expectations from the leaf economic spectrum. We show considerable trait plasticity in the woody shrub over microgeographic scales (<100 km), indicating it has the adaptive potential within a generation to functionally acclimate to a range of abiotic conditions. Since our study shrub is commonly used for restoration in southern Australia and local populations do not show strong genetic differentiation in functional traits, the potential risks of transferring seed across the broad environmental conditions are not likely to be a significant issue.

Transects that traverse substantial climate gradients are important tools for climate change research and allow questions on the extent to which phenotypic variation associates with climate, the link between climate and species distributions, and variation in sensitivity to climate change among biomes to be addressed. However, the potential limitations of individual transect studies have recently been highlighted. Here, we argue that replicating and networking transects, along with the introduction of experimental treatments, addresses these concerns. Transect networks provide cost-effective and robust insights into ecological and evolutionary adaptation and improve forecasting of ecosystem change. We draw on the experience and research facilitated by the Australian Transect Network to demonstrate our case, with examples, to clarify how population- and community-level studies can be integrated with observations from multiple transects, manipulative experiments, genomics, and ecological modeling to gain novel insights into how species and systems respond to climate change. This integration can provide a spatiotemporal understanding of past and future climate-induced changes, which will inform effective management actions for promoting biodiversity resilience.

Quantitative reconstructions of terrestrial climate are highly sought after but rare, particularly in Australia. Carbon isotope discrimination in plant leaves (Δleaf ) is an established indicator of past hydroclimate because the fractionation of carbon isotopes during photosynthesis is strongly influenced by water stress. Leaves of the evergreen tree Melaleuca quinquenervia have been recovered from the sediments of some perched lakes on North Stradbroke and Fraser Islands, south-east Queensland, eastern Australia. Here, we examine the potential for using M. quinquenervia ∆leaf as a tracer of past rainfall by analysing carbon isotope ratios (δ(13) C) of modern leaves. We firstly assess Δleaf variation at the leaf and stand scale and find no systematic pattern within leaves or between leaves due to their position on the tree. We then examine the relationships between climate and Δleaf for a 11-year time series of leaves collected in a litter tray. M. quinquenervia retains its leaves for 1-4 years; thus, cumulative average climate data are used. There is a significant relationship between annual mean ∆leaf and mean annual rainfall of the hydrological year for 1-4 years (i.e. 365-1460 days) prior to leaf fall (r(2)  = 0.64, P = 0.003, n = 11). This relationship is marginally improved by accounting for the effect of pCO2 on discrimination (r(2)  = 0.67, P = 0.002, n = 11). The correlation between rainfall and Δleaf , and the natural distribution of Melaleuca quinquenervia around wetlands of eastern Australia, Papua New Guinea and New Caledonia offers significant potential to infer past rainfall on a wide range of spatial and temporal scales., (© 2016 John Wiley & Sons Ltd.)

Leaf water contains naturally occurring stable isotopes of oxygen and hydrogen in abundances that vary spatially and temporally. When sufficiently understood, these can be harnessed for a wide range of applications. Here, we review the current state of knowledge of stable isotope enrichment of leaf water, and its relevance for isotopic signals incorporated into plant organic matter and atmospheric gases. Models describing evaporative enrichment of leaf water have become increasingly complex over time, reflecting enhanced spatial and temporal resolution. We recommend that practitioners choose a model with a level of complexity suited to their application, and provide guidance. At the same time, there exists some lingering uncertainty about the biophysical processes relevant to patterns of isotopic enrichment in leaf water. An important goal for future research is to link observed variations in isotopic composition to specific anatomical and physiological features of leaves that reflect differences in hydraulic design. New measurement techniques are developing rapidly, enabling determinations of both transpired and leaf water δ(18) O and δ(2) H to be made more easily and at higher temporal resolution than previously possible. We expect these technological advances to spur new developments in our understanding of patterns of stable isotope fractionation in leaf water., (© 2016 JohnWiley & Sons Ltd.)

Premise of the Study: Leaf venation is linked to physiological performance, playing a critical role in ecosystem function. Despite the importance of leaf venation, associated bundle sheath extensions (BSEs) remain largely unstudied. Here, we quantify plasticity in the spacing of BSEs over irradiance and precipitation gradients. Because physiological function(s) of BSEs remain uncertain, we additionally explored a link between BSEs and water use efficiency (WUE)., Methods: We sampled leaves of heterobaric trees along intracrown irradiance gradients in natural environments and growth chambers and correlated BSE spacing to incident irradiance. Additionally, we sampled leaves along a precipitation gradient and correlated BSE spacing to precipitation and bulk δ(13)C, a proxy for intrinsic WUE. BSE spacing was quantified using a novel semiautomatic method on fresh leaf tissue., Key Results: With increased irradiance or decreased precipitation, Liquidambar styraciflua decreased BSE spacing, while Acer saccharum showed little variation in BSE spacing. Two additional species, Quercus robur and Platanus occidentalis, decreased BSE spacing with increased irradiance in growth chambers. BSE spacing correlated with bulk δ(13)C, a proxy for WUE in L. styraciflua, Q. robur, and P. occidentalis leaves but not in leaves of A. saccharum., Conclusions: We demonstrated that BSE spacing is plastic with respect to irradiance or precipitation and independent from veins, indicating BSE involvement in leaf adaptation to a microenvironment. Plasticity in BSE spacing was correlated with WUE only in some species, not supporting a function in water relations. We discuss a possible link between BSE plasticity and life history, particularly canopy position.

Body size plays a critical role in mammalian ecology and physiology. Previous research has shown that many mammals became smaller during the Paleocene-Eocene Thermal Maximum (PETM), but the timing and magnitude of that change relative to climate change have been unclear. A high-resolution record of continental climate and equid body size change shows a directional size decrease of ~30% over the first ~130,000 years of the PETM, followed by a ~76% increase in the recovery phase of the PETM. These size changes are negatively correlated with temperature inferred from oxygen isotopes in mammal teeth and were probably driven by shifts in temperature and possibly high atmospheric CO(2) concentrations. These findings could be important for understanding mammalian evolutionary responses to future global warming.