Developmental stability and morphological differences in Plantago major L. leaves under contrasting environmental conditions

Abstract

In the present study, an additional combination of end‐points was applied on the natural populations of the common plantain, previously estimated using morphometric assays. Here, besides measuring developmental instability (DI), by determining the level of fluctuating asymmetry (FA) and the total amount of phenotypic variability (PV), we tried to distinguish the three natural populations under contrasting environmental conditions using the morphological data. Results obtained using both FA indices were the same; higher asymmetry levels in the reference than in the polluted environments were detected for leaf width, vein distances within a leaf and lobe length. The one‐way analysis of variance results revealed that there were significant differences in PV values among populations analysed for each character. When all leaf traits were considered together, the PV median value was significantly higher in Crni Lug leaves compared with leaves from other sites. The multivariate analysis of variance results revealed the significant effect of environment on both FA₄ and PV values. The component scores of first factor (PC1) were significantly different between the Karaburma and Crni Lug populations. Besides, component scores of both PC1 and PC2 were significantly different between the Zemun and Crni Lug samples. The stepwise discriminant functional analysis results allowed us to identify a set of four variables, with a sufficient discriminating ability (75%).     

Genetic and clonal structures of the tree species Tilia cordata mill. in remnants of ancient forests in Denmark

The insect-pollinated forest tree Tilia cordata Mill. grows today in small fragmented populations in Denmark and other western European countries but was, in prehistoric times, a dominating species and is considered an indicator species for ancient forest. The species is known to propagate both sexually and vegetatively, forming clonal groups. Few studies have been made on the species' population genetics and on how clonality affects the population structure. The aim of this study is to evaluate the Danish gene pool by estimating genetic diversity and differentiation, as well as through exhaustive sampling describe clonal structures in some of the populations. Genetic analysis was carried out using nine nuclear microsatellite markers in nine populations, of which four were exhaustively or partly exhaustively sampled. The markers showed a high degree of genetic diversity but low differentiation between populations, with no geographic-related structure. Clonal structures were found in eight out of the nine populations. In the exhaustively sampled populations, recruitment strategies included both sexual and clonal reproduction with indications that clonality may be enhanced by management and other disturbances.     

Whole organ, venation and epidermal cell morphological variations are correlated in the leaves of Arabidopsis mutants

Despite the large number of genes known to affect leaf shape or size, we still have a relatively poor understanding of how leaf morphology is established. For example, little is known about how cell division and cell expansion are controlled and coordinated within a growing leaf to eventually develop into a laminar organ of a definite size. To obtain a global perspective of the cellular basis of variations in leaf morphology at the organ, tissue and cell levels, we studied a collection of 111 non-allelic mutants with abnormally shaped and/or sized leaves, which broadly represent the mutational variations in Arabidopsis thaliana leaf morphology not associated with lethality. We used image-processing techniques on these mutants to quantify morphological parameters running the gamut from the palisade mesophyll and epidermal cells to the venation, whole leaf and rosette levels. We found positive correlations between epidermal cell size and leaf area, which is consistent with long-standing Avery's hypothesis that the epidermis drives leaf growth. In addition, venation parameters were positively correlated with leaf area, suggesting that leaf growth and vein patterning share some genetic controls. Positional cloning of the genes affected by the studied mutations will eventually establish functional links between genotypes, molecular functions, cellular parameters and leaf phenotypes.     

Developmental plasticity, morphological variation and evolvability: a multilevel analysis of morphometric integration in the shape of compound leaves

The structure of compound leaves provides flexibility for morphological change by variation in the shapes, sizes and arrangement of leaflets. Here, we conduct a multilevel analysis of shape variation in compound leaves to explore the developmental plasticity and evolutionary potential that are the basis of diversification in leaf shape. We use the methods of geometric morphometrics to study the shapes of individual leaflets and whole leaves in 20 taxa of Potentilla (sensu lato). A newly developed test based on the bootstrap approach suggests that uncertainty in the molecular phylogeny precludes firm conclusions whether there is a phylogenetic signal in the data on leaf shape. For variation among taxa, variation within taxa, as well as fluctuating asymmetry, there is evidence of strong morphological integration. The patterns of variation are similar across all three levels, suggesting that integration within taxa may act as a constraint on evolutionary change.     

Functional design space of single-veined leaves: role of tissue hydraulic properties in constraining leaf size and shape

BACKGROUND AND AIMS: Morphological diversity of leaves is usually quantified with geometrical characters, while in many cases a simple set of biophysical parameters are involved in constraining size and shape. One of the main physiological functions of the leaf is transpiration and thus one can expect that leaf hydraulic parameters can be used to predict potential morphologies, although with the caveat that morphology in turn influences physiological parameters including light interception and boundary layer thickness and thereby heat transfer and net photosynthesis. METHODS: An iterative model was used to determine the relative sizes and shapes that are functionally possible for single-veined leaves as defined by their ability to supply the entire leaf lamina with sufficient water to prevent stomatal closure. The model variables include the hydraulic resistances associated with vein axial and radial transport, as well as with water movement through the mesophyll and the leaf surface. KEY RESULTS: The four parameters included in the model are sufficient to define a hydraulic functional design space that includes all single-veined leaf shapes found in nature, including scale-, awl- and needle-like morphologies. This exercise demonstrates that hydraulic parameters have dissimilar effects: surface resistance primarily affects leaf size, while radial and mesophyll resistances primarily affect leaf shape. CONCLUSIONS: These distinctions between hydraulic parameters, as well as the differential accessibility of different morphologies, might relate to the convergent evolutionary patterns seen in a variety of fossil lineages concerning overall morphology and anatomical detail that frequently have evolved in linear and simple multi-veined leaves.     

Species limits and morphometric and environmental variation within the South Andean and Patagonian Mulinum spinosum species-group (Apiaceae-Azorelloideae)

Delimiting species is an important, but frequently difficult aspect of systematics that should be addressed using data from multiple sources. Here we combine morphometric analyses and environmental characteristics to delimit species in the South Andean and Patagonian taxonomically difficult species-group composed by Mulinum spinosum, M. echinus and M. leptacanthum (Apiaceae-Azorelloideae). Molecular phylogenies have shown that these three species form part of a polytomy together with other Mulinum species, and therefore these data are not useful for their delimitation. We include measurements of 25 morphometric variables from 163 herbarium specimens and perform univariate and multivariate principal component analysis (PCA) and discriminant analysis (DA) to establish the limits amongst the three mostly sympatric, morphologically similar, and phylogenetically unresolved species. We also use 19 bioclimatic and three geographic variables from localities of the specimens to infer environmental characteristics of the taxa and test their relation with morphological variation. Morphological evidence supports the inclusion of M. echinus within the morphologically and climatically variable M. spinosum, and rejects its recognition as a distinct taxon at any rank. On the contrary, M. leptacanthum is considered a morphologically distinct species, generally restricted to high altitude areas of the southern Andes with a cooler and wetter climate. Within the widespread M. spinosum, environmental gradients of precipitation and temperature relate to morphological gradients (e.g., in leaf and inflorescence sizes, in leaf acicularity, and in fruit-wing width). These last results showed that the large morphological variation in vegetative and reproductive characteristics of this species that grows in arid and semiarid habitats are related to regional climatic gradients that have probably been important in the evolution of this species' plasticity, diversification, and differentiation.     

Genetic signature of the natural gene pool of Tilia cordata Mill. in Lithuania: Compound evolutionary and anthropogenic effects

Tilia cordata Mill. is a valuable tree species enriching the ecological values of the coniferous‐dominated boreal forests in Europe. Following the historical decline, spreading of Tilia sp. is challenged by the elevated inbreeding and habitat fragmentation. We studied the geographical distribution of genetic diversity of Tilia cordata populations in Lithuania. We used 14 genomic microsatellite markers to genotype 543 individuals from 23 wild‐growing populations. We found that Tilia cordata retained high levels of genetic diversity (population F _IS = 0–0.15, H _o = 0.53–0.69, H _e = 0.56–0.75). AMOVA, Bayesian clustering, and Monmonier''s barrier detection indicate weak but significant differentiation among the populations (F _ST = 0.037***) into geographically interpretable clusters of (a) western Lithuania with high genetic heterogeneity but low genetic diversity, bottleneck effects, (b) relatively higher genetic diversity of Tilia cordata on rich and most soils of midland lowland, and (c) the most differentiated populations on poor soils of the coolest northeastern highland possessing the highest rare allele frequency but elevated inbreeding and bottleneck effects. Weak genetic differentiation among the Tilia cordata populations in Lithuania implies common ancestry, absence of strong adaptive gradients, and effective genetic exchange possible mediated via the riparian networks. A hypothesis on riparian networks as gene flow mediators in Tilia cordata was raised based on results of this study.     

Size and shape heritability in natural populations of Drosophila mediopunctata: temporal and microgeographical variation

‘Traditional morphometrics’ allows us to decompose morphological variation into its major independent sources, identifying them usually as size and shape. To compare and investigate the properties of size and shape in natural populations of Drosophila mediopunctata, estimating their heritabilities and analysing their temporal and microgeographic changes, we carried out collections on seven occasions in Parque Nacional do Itatiaia, Brazil. In one of these collections, we took samples from five different altitudes. Measurements were taken from wild caught inseminated females and up to three of their laboratory‐reared daughters. Through a principal component analysis, three major sources of variation were identified as due to size (the first one) and shape (the remaining two). The overall amount of variation among laboratory flies was about half of that observed among wild flies and this reduction was primarily due to size. Shape variation was about the same under natural and artificial conditions. A genetic altitudinal cline was detected for size and shape, although altitude explained only a small part of their variation. Differences among collections were detected both for size and shape in wild and laboratory flies, but no simple pattern emerged. Shape variation had high heritability in nature, close to or above 40% and did not vary significantly temporally. Although on the overall size heritability (18 ± 6%)was significant its estimates were not consistent along months – they were non‐significant in all but one month, when it reached a value of 51 ± 11%. Overall, this suggests that size and shape have different genetic properties. This revised version was published online in July 2006 with corrections to the Cover Date.     

竹子箨叶形态性状的种间差异及其与竹笋食味品质指标的关系

竹笋是大众喜爱的森林蔬菜,其食味品质存在种间差异,且出土前后品质存在明显的变异.箨叶是竹笋的重要器官,居于箨鞘顶端,是竹笋出土时最先感光部位,是否与竹笋食味品质变异密切相关?目前尚不清楚.为此,以同一地点、立地条件和人工干扰措施一致的6个属22个竹种为对象,分析了不同竹种箨叶形态性状和竹笋食味品质指标,为揭示竹笋食味品...     

Accumulation of particulate matter,heavy metals,and polycyclic aromatic hydrocarbons on the leaves of Tilia cordata Mill. in five Polish cities with different levels of air pollution

Urbanized areas are struggling with the problem of air pollution and as the number of people living in cities is increasing, the situation is likely to deteriorate. One of the most harmful pollutants is particulate matter (PM). Increased levels of PM in the atmosphere are likely to have a negative impact on human health. Phytoremediation technology could be a solution. It involves plants acting as bio-filters by accumulating particles on, and in the leaves, thus removing the particles from the atmosphere. This study investigates the accumulation of PM including heavy metals (HMs) and polycyclic aromatic hydrocarbons (PAHs), on the foliage of small-leaved lime (Tilia cordata Mill.) in five Polish cities. There were significantly different PM amounts found in the trees between the cities which related to the different quantities of PM in the atmosphere at these cities. Significant differences were found between cities for the amounts of the different particulate size fractions, and the HMs and PAHs in leaves. Strong winds reduced the amount of PM on leaves, especially the smallest fractions, but no relationship with precipitation was found. The results suggest that T. cordata improves the air quality in cities and can be used as an effective bioindicator for PM air pollution.     

HPLC fingerprints combined with principal component analysis,hierarchical cluster analysis and linear discriminant analysis for the classification and differentiation of Peganum sp. indigenous to China

Introduction – Seeds of wild Peganum harmala Linn., P. multisectum (Maxim) Bobr., P. nigellastrum Bunge and a probable indeterminate species, herein referred to as P. variety, are commonly used in Chinese medicine. These seeds cannot be differentiated based on morphology. Objective – Seeds of P. harmala Linn., P. multisectum (Maxim) Bobr., P. nigellastrum Bunge and P. variety were collected in different provinces in China and their HPLC profiles were recorded for statistical analysis and pattern recognition. Methodology – HPLC chromatograms of seed extracts were recorded under the same conditions. Individual HPLC chromatograms for each species were evaluated against the mean chromatogram for the same species generated using a similarity evaluation computer program. Data from chromatographic fingerprints were also processed using principal component analysis (PCA), hierarchical cluster analysis (HCA) and linear discriminant analysis (LDA). Results – The Peganum sp. seed extracts had similar HPLC fingerprints but with some inter‐specific differences. The chromatographic fingerprints combined with PCA, HCA and LDA could distinguish the seeds of the different species of Peganum investigated. Conclusion – HPLC fingerprints can be used to authenticate and differentiate the seeds of three different species of genus Peganum indigenous to China. The results indicated that the unidentified P. variety might indeed be a new species or variety. Copyright © 2009 John Wiley & Sons, Ltd.     

Quantitative analysis of cell division in leaves: methods,developmental patterns and effects of environmental conditions

In planta quantitative studies of cell cycle are necessary for examining the role of cell division in the response of plants to environmental conditions and to analyse the behaviour of transformed plants in this context. We present and discuss non-intrusive kinematic methods which allow estimating the duration of cell cycle with a high spatial resolution in the leaf. Different methods are proposed and discussed for monocotyledons and dicotyledons, and compared with methods involving the use of chemicals. In monocotyledon leaves, cell division is restricted to a limited zone near the leaf insertion point, twice as long in the mesophyll as in the epidermis. In dicotyledons, cell division occurs in the whole leaf with a uniform and constant cell cycle duration for a determinate number of cell cycles, representing about half of leaf development. Over several experiments, this number is well conserved in a given leaf zone in the absence of stresses, but larger near the leaf base than near the leaf tip. After that, cell cycle duration increases because cells are progressively blocked in G1 while the durations of S-G2-M phases do not change with time. Leaf temperature affects neither the distribution of nuclei in each phase of the cycle nor the number of cell cycles in a leaf. Water or light deficits both cause a partial blockage of nuclei in G1 during the stress only, thereby increasing cell cycle duration and decreasing final cell number. These results suggest that a strong developmental programme drives cell division in leaves, so a simple framework allows analysis of temporal patterns, of spatial gradients and of the effect of environmental conditions.     

Allozymic and morphometric analysis of populations in the Zygaena purpuralis complex (Lepidoptera, Zygaenidae)

In Central Europe and Asia Minor Zygaena purpuralis and Z. minos are two sibling species exhibiting a considerable geographic variation in several features. In this study we examine some populations from the Central and Southern Apennines (Italy) by comparing multivariate analyses of individual allozymic multilocus profiles and of individual morphometric profiles of male genitalia in order to assess their genetic and phenetic relationships.
Results indicate that populations of Z. purpuralis in the Apennines behave in rather different ways with respect to the occurrence of gene exchange with other gene pools. No gene flow appears to occur between two geographically close southern populations; however, extensive introgression, revealable at both the allozymic and morphometric level, seems to occur between some purpuralis and minos -like sympatric populations from Abruzzi. Evolutionary relationships among these peninsular Italy populations are discussed with reference to historical factors, that is, the occurrence of multiple Pleistocene refugia, which could have determined different degrees of isolation among gene pools. This situation is rather different from that previously outlined for Central Europe and present results suggest that the Z. purpuralis complex might include more than two biological species.     

Morphological and genetic analysis of fish of a Carassius complex (Cyprinidae) in Lake Kasumigaura with reference to the taxonomic status of two all-female triploid morphs

The classification of the Carassius complex (Cyprinidae) including all-female triploids, called ginbuna in Japanese, is so confused that three sympatric morphs of crucian carp in Lake Kasumigaura are categorized into two different subspecies within a species. We examined them in order to explain the coexistence of more than one subspecies and determine the founder of the triploid lineages in the crucian carp fauna in the lake. Principal component analysis proved that the three sympatric morphs had a morphometric basis distinguishable from each other. Ploidy was determined by flow cytometry which showed triploids in two morphs and diploids in the other morph. Stepwise discriminant analysis using only meristic characteristics could separate the diploids from the triploids. Phylogenetic analysis using mitochondrial DNA inferred two lineages in which one was composed of a triploid morph and the other was a diploid–triploid mixture. Disagreement between the taxonomic status and the phylogenetic status is explicable by assuming that the triploids in the Carassius complex had independent origins leading to the different subspecies. C. auratus langsdorfii appears to show genetic complexities that traditional taxonomic classification can not unravel.  © 2003 The Linnean Society of London, Biological Journal of the Linnean Society , 2003, 79 , 351–357.     

贵州省典型植烟生态区域根际土壤微生物群落多样性

土壤微生物是土壤的重要组成部分,它对土壤肥力的形成和植物营养的转化起着积极的作用。采用16S V4区高通量测序等方法对贵州省不同生态区域典型植烟土壤根际微生物群落进行分析研究,从根际土壤微生物群落特征上诠释不同生态区域典型土壤根际微生物群落的信息差异。结果表明:贵州省不同生态区域弱酸和酸性土壤占70.06%,有机碳平均2.24%,有机碳含量总体偏高,全氮与有机碳含量具有一定相关性;根际土壤微生物的OTUs数量差异明显,呈现总体是西部高于东部,南部高于北部;微生物属水平物种丰度差异显示,土壤类型对微生物群落丰度影响显著,而在土壤功能微生物上黔北区域的变形菌门(Proteobacteria)和放线菌门(Actinobacteria)两个门的微生物相对丰度最高,而在黔中中部区域变形菌门的丰度显著高于放线菌门的丰度。黔西南和黔中中部区域土壤微生物属水平优势种群显著高于其他地区,种群平衡度不高,具有土传病害发生的潜在危险。通过贵州省不同生态区域土壤微生物根际微生物信息分析与研究,为后续土壤生物修复奠定了良好的研究基础。     

不同生境白屈菜(Chelidonium majus)生活史型特征及其与不同器官单宁、黄酮、生物碱含量的关系

运用主成分分析法,对不同生境条件下白屈菜植物生活史型相关的营养生长（vegetative growth,V）、有性生长（sexual growth,S）和克隆生长（clonal growth,C）的主成分得分及比例进行定量计算,并以此为基础研究了生活史型特征参数及次生代谢产物单宁含量、黄酮含量与生物碱含量的相关关系,目的是为植物活性成分环境定向诱导培育提供实验依据。研究结果表明：（1）对于不同光照状况的全光照（空地）、70%光照（榆树林内）和40%光照下（白扦林内）的9个样地白屈菜生活史型划分发现,全光照下白屈菜种群生活史型为V0.34S0.41C0.25,为SV生活史型,空地为DE（Disturbed but still Excellent）生境;榆树林下和白扦林下的白屈菜生活史型分别可表示为V0.28S0.38C0.34和V0.27S0.40C0.33,均为SC生活史型,榆树林下和白扦林下为DF（Disturbed and Fragile）生境。（2）白屈菜植株次生代谢产物（单宁、黄酮和生物碱）含量,空地均低于榆树林下和白扦林下生境。各样地白屈菜不同器官单宁含量：叶片（种（根（茎;黄酮含量：种（叶片（根（茎;生物碱含量：叶片（根（茎,各样地白屈菜茎和根的生物碱含量无明显差异,空地白屈菜叶片中生物碱含量低于榆树林下和白扦林下白屈菜植株59%～56.7%。（3）白屈菜生活史型与次生代谢产物（单宁、黄酮和生物碱）含量相关性分析结果中,显著的线性关系显示,白屈菜次生代谢产物（单宁、黄酮和生物碱）含量与营养生长和有性生长成负相关,与克隆生长成正相关。实验结果表明,较于空地的DE生境,林（榆树和白扦）下的DF生境条件差,使白屈菜向C型转变,同时也促进了次生代谢产物（单宁、黄酮和生物碱）的积累。结果可以为野生植物的人工定向培育中生境选择和目的活性成分定向累积提供基于形态学的评价方法和理论。     

中国亚热带不同菌根树种的根叶形态学性状特征与生长差异: 以江西新岗山为例

探究植物功能性状的种内和种间变异不仅有助于揭示植物对环境的适应, 也能够反映植物的生态策略, 但不同菌根类型树木生长过程中根叶形态学功能性状的适应策略仍有待探究。本研究依托中国亚热带森林生物多样性与生态系统功能实验研究平台(BEF-China)选取7种丛枝菌根(AM)树木和7种外生菌根(EM)树木的纯林, 测定各个树种的比叶面积、叶干物质含量、比根长、根系直径、树高生长速率、地径生长速率及细根生物量等根叶形态学功能性状和生长指标, 探讨了两种菌根类型树种间的根叶形态学特征的差异。结果表明: 与AM树种相比, EM树种具有较小的比叶面积、吸收根平均直径和生长速率, 但具有更大的叶干物质含量; 两种菌根树种之间的比根长和细根生物量无显著差异。比叶面积、叶干物质含量、树高生长速率、地径生长速率和细根生物量等功能性状及生长指标在不同菌根类型、树种及二者的交互作用中均存在显著差异; 且树种、根功能型、菌根类型及三者之间的交互作用均对根功能性状有显著影响。EM树种地上指标的种内变异均大于种间变异, 而AM树种地上指标的种内和种间变异程度类似; 但两种菌根树种细根生物量的种间变异均大于种内变异。尽管两种菌根树种地上部分生长速率较快通常表现为较低的叶干物质含量, 但AM树种通常拥有较高的吸收根比根长, 而EM树种拥有较粗的运输根平均直径。吸收根比根长越低, 两类菌根树种的细根生物量就越多。由此可见, 根叶功能性状对植物地上部分的生长具有一定的协同效应, 其中运输根主要在EM树种地上生长过程中发挥重要作用, 吸收根主要与AM树种的地上部分生长有关; 但两类菌根树种的地下细根生物量均与吸收根有关。     

Patterns of leaf asymmetry changes in Plantago major L. (ssp. major) natural populations exposed to different environmental conditions

In the present study, developmental stability of leaf traits was examined in three natural populations of Plantago major L. (ssp. major), representing two polluted environments (Karaburma and Zemun) and an unpolluted area (Crni Lug). Developmental stability was assessed as fluctuating asymmetry (FA). The magnitude of FA is believed to reflect differences in the ability of individuals to buffer their development in natural populations. We hypothesized that there are differences within characters and among characters in response to environmental conditions. Significant patterns of asymmetry correlations and asymmetry changes were detected both within characters and between characters. The manova results revealed a significant effect of individual and a significant individual × environment interaction on actual asymmetry (logL_i ? logR_i) and on the amount of asymmetry |(logL_i ? logR_i)| for leaf width and vein distances within a leaf. Over time, statistically significant and positive correlations of the FA values were detected for each trait separately per sample (population). For both leaf traits, there were differences for (logL_i ? logR_i) and |(logL_i ? logR_i)| asymmetry values among individuals within samples in response to yearly variations. Statistically significant and negative correlations for (logL_i ? logR_i) versus |(logL_i ? logR_i)| asymmetries were detected for both leaf traits. In summary, our results highlight the importance of differences in the ability of individuals to buffer their development under different environmental conditions and point to the concept that developmental stability is character specific.     

Global quantification of contrasting leaf life span strategies for deciduous and evergreen species in response to environmental conditions

Aim Species with deciduous and evergreen leaf habits typically differ in leaf life span (LLS). Yet quantification of the response of LLS, within each habit, to key environmental conditions is surprisingly lacking. The aim of this study is to quantify LLS strategies of the two leaf habits under varying temperature, moisture and nutrient conditions, using a global database. We hypothesize that deciduous LLS reflects the length of the growing season, avoiding unfavourable conditions regardless of the cause. Evergreen species adjust to unfavourable periods and amortize lower net carbon gains over several growing seasons, with increasing LLS associated with increasingly short favourable versus unfavourable season lengths. Location Global. Methods Data on LLS and environmental variables were compiled from global datasets for 189 deciduous and 506 evergreen species across 83 study locations. Individual and combined effects of measures of seasonality of temperature, water and nutrient availability on length of the growing season and on LLS were quantified using linear mixed models. The best models for predicting LLS were obtained using Akaike's information criterion (AIC) and ΔAIC. Results The LLS of deciduous and evergreen species showed opposite responses to changes in environmental conditions. Under unfavourable conditions, deciduous LLS decreases while evergreen LLS increases. A measure of temperature alone was the best predictor of the growing season. The LLS of deciduous species was independent of environmental conditions after expressing LLS in relation to the number of growing seasons. Evergreen species, on the other hand, adjusted to unfavourable conditions by increasing LLS up to four growing seasons. Contrary to expectations, variation in LLS was best explained solely by temperature, instead of by combined measures of temperature, moisture and nutrient availability. Shifts in the photosynthesis to respiration balance might provide a physiological explanation. Main conclusions Temperature, and not drought or nutrient availability, is the primary driver of contrasting responses of LLS for different leaf habit types.