FLOWERING LOCUS T-mediated thermal signalling regulates age-dependent inflorescence development in Arabidopsis thaliana.

Many plants show strong heteroblastic changes in the shape and size of organs as they transition from juvenile to reproductive age. Most attention has been focused on heteroblastic development in leaves, but we wanted to understand heteroblastic changes in reproductive organ size. We therefore studied the progression of reproductive development in the model plant Arabidopsis thaliana, and found strong reductions in the size of flowers, fruit, seed, and internodes during development. These did not arise from correlative inhibition by older fruits, or from changes in inflorescence meristem size, but seemed to stem from changes in the size of floral organ primordia themselves. We hypothesized that environmental conditions might influence this heteroblastic pattern and found that the ambient temperature during organ initiation strongly influences organ size. We show that this temperature-dependent heteroblasty is dependent on FLOWERING LOCUS T (FT)-mediated signal integration, adding to the repertoire of developmental processes regulated by this pathway. Our results demonstrate that rising global temperatures will not affect just fertility, as is widely described, but also the size and seed number of fruits produced. However, we also show that such effects are not hard-wired, and that selective breeding for FT expression during reproductive development could mitigate such effects.

Expression divergence of expansin genes drive the heteroblasty in Ceratopteris chingii.

BACKGROUND: Sterile-fertile heteroblasty is a common phenomenon observed in ferns, where the leaf shape of a fern sporophyll, responsible for sporangium production, differs from that of a regular trophophyll. However, due to the large size and complexity of most fern genomes, the molecular mechanisms that regulate the formation of these functionally different heteroblasty have remained elusive. To shed light on these mechanisms, we generated a full-length transcriptome of Ceratopteris chingii with PacBio Iso-Seq from five tissue samples. By integrating Illumina-based sequencing short reads, we identified the genes exhibiting the most significant differential expression between sporophylls and trophophylls. RESULTS: The long reads were assembled, resulting in a total of 24,024 gene models. The differential expressed genes between heteroblasty primarily involved reproduction and cell wall composition, with a particular focus on expansin genes. Reconstructing the phylogeny of expansin genes across 19 plant species, ranging from green algae to seed plants, we identified four ortholog groups for expansins. The observed high expression of expansin genes in the young sporophylls of C. chingii emphasizes their role in the development of heteroblastic leaves. Through gene coexpression analysis, we identified highly divergent expressions of expansin genes both within and between species. CONCLUSIONS: The specific regulatory interactions and accompanying expression patterns of expansin genes are associated with variations in leaf shapes between sporophylls and trophophylls.

Interspecific morphological variation in Juglandoideae resting bud organization: a winter's tale?

BACKGROUND AND AIMS: Dormant resting buds are frequently regarded as static units, with protective cataphylls on the outside and embryonic foliage leaves on the inside. How the presence of cataphylls influences the dynamic, cyclical, annually repeating sequence of leaf forms that a resting bud gives rise to has rarely been interrogated. To examine the connection between dormant structure and growing-season development, we compare the complete seasonal heteroblastic sequence of leaf forms of six species of temperate Juglandaceae with distinctly different vegetative resting bud structures. These include buds with cataphylls; buds without cataphylls; and buds with caducous cataphylls that are lost before the onset of winter. METHODS: In a common garden setting over a 7-month growing season, the dimensions of 2249 individual vegetative metamers were tracked from first exposure to abscission along the shoots of saplings and mature trees. The timing of metamer initiation within terminal buds was investigated using micro-CT scanning. Character state transitions of resting bud types were estimated using a phylogenetic tree of Juglandaceae. KEY RESULTS: The presence of cataphylls within a heteroblastic sequence is associated with a single cohort of foliage leaves that flush and abscise synchronously. This growing pattern is highly determinate, with next year's terminal-bud cataphylls already initiated before spring leaf out. In contrast, in sequences without cataphylls, shorter-lived foliage leaves appear and abscise in a staggered fashion. Despite these differences in leaf demography, all examined heteroblastic sequences produce a series of small, caducous leaf forms that precede terminal bud set. CONCLUSIONS: The ubiquity of caducous leaf forms in Juglandoideae may point to the importance of shoot tip protection far beyond the dormant season. In addition, the presence or absence of cataphylls in resting buds is indicative of distinct shoot ontogenetic patterns, and functional strategies, in summer.

Shoot dimorphism enables Sequoia sempervirens to separate requirements for foliar water uptake and photosynthesis.

PREMISE: Trees in wet forests often have features that prevent water films from covering stomata and inhibiting gas exchange, while many trees in drier environments use foliar water uptake to reduce water stress. In forests with both wet and dry seasons, evergreen trees would benefit from producing leaves capable of balancing rainy-season photosynthesis with summertime water absorption. METHODS: Using samples collected from across the vertical gradient in tall redwood (Sequoia sempervirens) crowns, we estimated tree-level foliar water uptake and employed physics-based causative modeling to identify key functional traits that determine uptake potential by setting hydraulic resistance. RESULTS: We showed that Sequoia has two functionally distinct shoot morphotypes. While most shoots specialize in photosynthesis, the axial shoot type is capable of much greater foliar water uptake, and its within-crown distribution varies with latitude. A suite of leaf surface traits cause hydraulic resistance, leading to variation in uptake capacity among samples. CONCLUSIONS: Shoot dimorphism gives tall Sequoia trees the capacity to absorb up to 48 kg H2 O h-1 during the first hour of leaf wetting, ameliorating water stress while presumably maintaining high photosynthetic capacity year round. Geographic variation in shoot dimorphism suggests that plasticity in shoot-type distribution and leaf surface traits helps Sequoia maintain a dominate presence in both wet and dry forests.

Exploring evolutionary adaptations of leaf heteroblasty in subspecific taxa of Hawaiian Chenopodium oahuense.

PREMISE: Chenopodium oahuense is a polymorphic Hawaiian endemic plant inhabiting several xeric habitats. Considerable variability in leaf morphology has made comprehensively describing its diversity difficult. Chenopodium oahuense subsp. ilioense is differentiated from C. oahuense subsp. oahuense by smaller, less-lobed, succulent leaves, smaller seeds, and prostrate to scandent habit. The lacking quantification of leaf shape, succulence, and previously unknown heteroblastic leaf transition in C. oahuense subsp. ilioense complicates the morphological boundaries separating subspecies. METHODS: This study used landmark analyses, elliptical Fourier descriptors (EFDs), and traditional shape descriptors measured from 1585 greenhouse-grown plant leaves collected over 18 weeks. Principal component analyses visualized correlations in leaf shape, and linear discriminant analyses predicted classifications, either subspecific or heteroblastic. RESULTS: Identity determination and heteroblastic change visualization were limited in landmark analyses. On the basis of EFDs and shape descriptors, C. oahuense subsp. ilioense was determined to be morphologically differentiated from C. oahuense subsp. oahuense with the Pu'u Ka Pele population as intermediate. The EFDs depicted heteroblastic change, predominantly in lobing. All analyses were restricted in correctly attributing a leaf to the week collected. Shape descriptors generally represented significant heteroblastic change over the growth period. CONCLUSIONS: These analyses support significant differentiation between the subspecies, particularly from shape descriptors. Furthermore, we quantified the morphological intermediacy of the Pu'u Ka Pele population. Results suggest this population could be the result of incomplete lineage sorting or a recent hybridization of the two subspecies. Hawaiian Chenopodium is a polymorphic lineage notable for future research in adaptive radiations, phenotypic plasticity, and heteroblasty.

CURLED LATER1 encoding the largest subunit of the Elongator complex has a unique role in leaf development and meristem function in rice.

The Elongator complex, which is conserved in eukaryotes, has multiple roles in diverse organisms. In Arabidopsis thaliana, Elongator is shown to be involved in development, hormone action and environmental responses. However, except for Arabidopsis, our knowledge of its function is poor in plants. In this study, we initially carried out a genetic analysis to characterize a rice mutant with narrow and curled leaves, termed curled later1 (cur1). The cur1 mutant displayed a heteroblastic change, whereby the mutant leaf phenotype appeared specifically at a later adult phase of vegetative development. The shoot apical meristem (SAM) was small and the leaf initiation rate was low, suggesting that the activity of the SAM seemed to be partially reduced in cur1. We then revealed that CUR1 encodes a yeast ELP1-like protein, the largest subunit of Elongator. Furthermore, disruption of OsELP3 encoding the catalytic subunit of Elongator resulted in phenotypes similar to those of cur1, including the timing of the appearance of mutant phenotypes. Thus, Elongator activity seems to be specifically required for leaf development at the late vegetative phase. Transcriptome analysis showed that genes involved in protein quality control were highly upregulated in the cur1 shoot apex at the later vegetative phase, suggesting the restoration of impaired proteins probably produced by partial defects in translational control due to the loss of function of Elongator. The differences in the mutant phenotype and gene expression profile between CUR1 and its Arabidopsis ortholog suggest that Elongator has evolved to play a unique role in rice development.

Green greenhouse: leaf enclosure for fruit development of an androdioecious vine, Schizopepon bryoniifolius.

Individual plants can produce leaves that differ substantially in size, morphology and many other traits. However, leaves that play a specific role in reproduction have rarely been reported. Here, we report leaves specialized to enclose fruit clusters and enhance seed production in an annual vine, Schizopepon bryoniifolius. Enclosure leaves were produced at the end of the growing season in late autumn. They were different in greenness and structure from other leaves. Under solar radiation, the ambient temperature inside an intact enclosure was up to 4.6°C higher than that near a fruit cluster whose enclosure leaves had been removed. We found that enclosures were thicker at colder sites. Removal of enclosing leaves negatively affected fruit survival and/or growth, but we could not identify the exact mechanism. The results suggested that enclosures allow the plant to produce seeds under the cold weather the plant encounters at the end of its life. Vegetative and reproductive traits of plants have usually been studied separately. This study indicates how they can dynamically interact, as shown by an examination of associations among leaf and reproductive trait changes according to life stages.

Phyllotaxis: is the golden angle optimal for light capture?

Phyllotactic patterns are some of the most conspicuous in nature. To create these patterns plants must control the divergence angle between the appearance of successive organs, sometimes to within a fraction of a degree. The most common angle is the Fibonacci or golden angle, and its prevalence has led to the hypothesis that it has been selected by evolution as optimal for plants with respect to some fitness benefits, such as light capture. We explore arguments for and against this idea with computer models. We have used both idealized and scanned leaves from Arabidopsis thaliana and Cardamine hirsuta to measure the overlapping leaf area of simulated plants after varying parameters such as leaf shape, incident light angles, and other leaf traits. We find that other angles generated by Fibonacci-like series found in nature are equally optimal for light capture, and therefore should be under similar evolutionary pressure. Our findings suggest that the iterative mechanism for organ positioning itself is a more likely target for evolutionary pressure, rather than a specific divergence angle, and our model demonstrates that the heteroblastic progression of leaf shape in A. thaliana can provide a potential fitness benefit via light capture.

Multiscale quantification of morphodynamics: MorphoLeaf software for 2D shape analysis.

A major challenge in morphometrics is to analyse complex biological shapes formed by structures at different scales. Leaves exemplify this challenge as they combine differences in their overall shape with smaller shape variations at their margin, leading to lobes or teeth. Current methods based on contour or on landmark analysis are successful in quantifying either overall leaf shape or leaf margin dissection, but fail in combining the two. Here, we present a comprehensive strategy and its associated freely available platform for the quantitative, multiscale analysis of the morphology of leaves with different architectures. For this, biologically relevant landmarks are automatically extracted and hierarchised, and used to guide the reconstruction of accurate average contours that properly represent both global and local features. Using this method, we establish a quantitative framework of the developmental trajectory of Arabidopsis leaves of different ranks and retrace the origin of leaf heteroblasty. When applied to different mutant forms, our method can contribute to a better understanding of gene function, as we show here for the role of CUC2 during Arabidopsis leaf serration. Finally, we illustrate the wider applicability of our tool by analysing hand morphometrics.

Leaf heteroblasty in Passiflora edulis as revealed by metabolic profiling and expression analyses of the microRNAs miR156 and miR172.

BACKGROUND AND AIMS: Juvenile-to-adult phase transition is marked by changes in leaf morphology, mostly due to the temporal development of the shoot apical meristem, a phenomenon known as heteroblasty. Sugars and microRNA-controlled modules are components of the heteroblastic process in Arabidopsis thaliana leaves. However, our understanding about their roles during phase-changing in other species, such as Passiflora edulis, remains limited. Unlike Arabidopsis, P. edulis (a semi-woody perennial climbing vine) undergoes remarkable changes in leaf morphology throughout juvenile-to-adult transition. Nonetheless, the underlying molecular mechanisms are unknown. METHODS: Here we evaluated the molecular mechanisms underlying the heteroblastic process by analysing the temporal expression of microRNAs and targets in leaves as well as the leaf metabolome during P. edulis development. KEY RESULTS: Metabolic profiling revealed a unique composition of metabolites associated with leaf heteroblasty. Increasing levels of glucose and α-trehalose were observed during juvenile-to-adult phase transition. Accumulation of microRNA156 (miR156) correlated with juvenile leaf traits, whilst miR172 transcript accumulation was associated with leaf adult traits. Importantly, glucose may mediate adult leaf characteristics during de novo shoot organogenesis by modulating miR156-targeted PeSPL9 expression levels at early stages of shoot development. CONCLUSIONS: Altogether, our results suggest that specific sugars may act as co-regulators, along with two microRNAs, leading to leaf morphological modifications throughout juvenile-to-adult phase transition in P. edulis.

Plasticity of phenotype and heteroblasty in contrasting populations of Acacia koa.

BACKGROUND AND AIMS: Heteroblastic plant species, whose morphology or growth habit changes suddenly during development, offer unique opportunities to investigate the role of selection in canalizing development or increasing the adaptive importance of plasticity. Leaf forms of the Hawaiian tree Acacia koa (koa) change morphologically and physiologically during the first year of growth, providing time to study abiotic factors influencing transition rates relative to other Acacia species. METHODS: The roles of light and water availability in triggering transition to the mature leaf form in contrasting (wet/dry) ecotypes of koa were investigated using a novel modelling technique to distinguish between chronological and ontogenetic controls in triggering transition. A light quality treatment was included to test interactions of heterophylly (the presence of multiple leaf forms) with heteroblastic processes on the resulting phenotype at transition. KEY RESULTS: Increased light intensity increased transition rates, but reduced red to far-red light (R:FR) ratios did not affect transition rates, solidifying the current paradigm of heteroblasty. However, evidence was found for earlier transition ontogenetically under water stress, which is not part of the current paradigm and could differentiate the role of heteroblasty in some Acacia species versus other heteroblastic species. Ecotypic responses also indicate that plasticity of development could vary across koa's range and the adaptive significance of heteroblasty could be marginalized or amplified dependent on the disparate selective pressures present across koa's range. CONCLUSIONS: The use of novel survival functions and a species with an elongated transition time helped to elucidate abiotic modifiers of ontogenetic trajectories. Differences in ontogenetic trajectories between contrasting ecotypes suggest that ongoing climate and land use change will have non-uniform effects on koa regeneration and establishment dynamics across its range.

Leaf Form Evolution in Viburnum Parallels Variation within Individual Plants.

Few studies have critically evaluated how morphological variation within individual organisms corresponds to variation within and among species. Subindividual variation in plants facilitates such studies because their indeterminate modular growth generates multiple serially homologous structures along growing axes. Focusing on leaf form, we evaluate how subindividual trait variation relates to leaf evolution across Viburnum, a clade of woody angiosperms. In Viburnum we infer multiple independent origins of wide/lobed leaves with toothed margins from ancestors with elliptical, smooth-margined leaves. We document leaf variation along the branches of individual plants of 28 species and among populations across the wide range of Viburnum dentatum. We conclude that when novel leaf forms evolved in Viburnum, they were intercalated at the beginning of the seasonal leaf sequence, which then generated a repeated spectrum of leaf forms along each branch (seasonal heteroblasty). We hypothesize that the existence of such a spectrum then facilitated additional evolutionary shifts, including reversions to more ancestral forms. We argue that the recurrent production of alternative phenotypes provides opportunities to canalize the production of particular forms and that this phenomenon has played an important role in generating macroscale patterns.

Heteroblasty in epiphytic bromeliads: functional implications for species in understorey and exposed growing sites.

Background and Aims: The functional relevance of heteroblasty, an abrupt morphological change in the ontogeny of a considerable number of angiosperm species, is still largely unresolved. During the ontogeny of many epiphytic Tillandsioids (Bromeliaceae), such a change occurs when small individuals transform into larger, tank-forming individuals that are capable of external water storage. Apart from its fundamental effect on plant water relations, the associated transition from narrow to broader leaves also affects plant architecture. The morphological changes and their effect on light interception may be especially relevant for heteroblastic species in the moist understorey, which are expected to be limited primarily by light. Methods: A functional structural plant model (Yplant) was used to construct digital replicas of atmospheric and tank-forming individuals of four species, two of them naturally growing in exposed conditions and two occurring in understorey sites. This allowed the determination of leaf display efficiencies as well as a systematic analysis of leaf architectural traits and their effect on light interception. Key Results: Modifying existing plant morphologies showed that broader leaves cause more self-shading within the plant. This supports the hypothesis that species from the light-limited understorey benefit from the early atmospheric life form through increased light capture. Modelling plant morphology that continuously followed the ontogenetic trajectories of the leaf architectural traits revealed that the rising total leaf number in atmospheric individuals constantly increased self-shading. Therefore, at a certain ontogenetic stage, a tipping point was reached when the tank form was even favourable in terms of light capture as it was associated with fewer leaves. Conclusions: The effects of changes in leaf morphology and leaf architecture on plant light capture may explain the common occurrence of heteroblastic species in the understorey of Neotropical forests, which does not negate a simultaneous positive effect of heteroblasty on plant water relations.

Transcriptomic analysis suggests a key role for SQUAMOSA PROMOTER BINDING PROTEIN LIKE, NAC and YUCCA genes in the heteroblastic development of the temperate rainforest tree Gevuina avellana (Proteaceae).

Heteroblasty, the temporal development of the meristem, can produce diverse leaf shapes within a plant. Gevuina avellana, a tree from the South American temperate rainforest shows strong heteroblasty affecting leaf shape, transitioning from juvenile simple leaves to highly pinnate adult leaves. Light availability within the forest canopy also modulates its leaf size and complexity. Here we studied how the interaction between the light environment and the heteroblastic progression of leaves is coordinated in this species. We used RNA-seq on the Illumina platform to compare the range of transcriptional responses in leaf primordia of G. avellana at different heteroblastic stages and growing under different light environments. We found a steady up-regulation of SQUAMOSA PROMOTER BINDING PROTEIN LIKE (SPL), NAC, YUCCA and AGAMOUS-LIKE genes associated with increases in age, leaf complexity, and light availability. In contrast, expression of TCP, TPR and KNOTTED1 homeobox genes showed a sustained down-regulation. Additionally, genes involved in auxin synthesis/transport and jasmonate activity were differentially expressed, indicating an active regulation of processes controlled by these hormones. Our large-scale transcriptional analysis of the leaf primordia of G. avellana sheds light on the integration of internal and external cues during heteroblastic development in this species.

Identifying developmental phases in the Arabidopsis thaliana rosette using integrative segmentation models.

The change in leaf size and shape during ontogeny associated with heteroblastic development is a composite trait for which extensive spatiotemporal data can be acquired using phenotyping platforms. However, only part of the information contained in such data is exploited, and developmental phases are usually defined using a selected organ trait. We here introduce new methods for identifying developmental phases in the Arabidopsis rosette using various traits and minimum a priori assumptions. A pipeline of analysis was developed combining image analysis and statistical models to integrate morphological, shape, dimensional and expansion dynamics traits for the successive leaves of the Arabidopsis rosette. Dedicated segmentation models called semi-Markov switching models were built for selected genotypes in order to identify rosette developmental phases. Four successive developmental phases referred to as seedling, juvenile, transition and adult were identified for the different genotypes. We show that the degree of covering of the leaf abaxial surface with trichomes is insufficient to define these developmental phases. Using our pipeline of analysis, we were able to identify the supplementary seedling phase and to uncover the structuring role of various leaf traits. This enabled us to compare on a more objective basis the vegetative development of Arabidopsis mutants.

Species diversity vs. morphological disparity in the light of evolutionary developmental biology.

BACKGROUND: Two indicators of a clade's success are its diversity (number of included species) and its disparity (extent of morphospace occupied by its members). Many large genera show high diversity with low disparity, while others such as Euphorbia and Drosophila are highly diverse but also exhibit high disparity. The largest genera are often characterized by key innovations that often, but not necessarily, coincide with their diagnostic apomorphies. In terms of their contribution to speciation, apomorphies are either permissive (e.g. flightlessness) or generative (e.g. nectariferous spurs). SCOPE: Except for Drosophila, virtually no genus among those with the highest diversity or disparity includes species currently studied as model species in developmental genetics or evolutionary developmental biology (evo-devo). An evo-devo approach is, however, potentially important to understand how diversity and disparity could rapidly increase in the largest genera currently accepted by taxonomists. The most promising directions for future research and a set of key questions to be addressed are presented in this review. CONCLUSIONS: From an evo-devo perspective, the evolution of clades with high diversity and/or disparity can be addressed from three main perspectives: (1) evolvability, in terms of release from previous constraints and of the presence of genetic or developmental conditions favouring multiple parallel occurrences of a given evolutionary transition and its reversal; (2) phenotypic plasticity as a facilitator of speciation; and (3) modularity, heterochrony and a coupling between the complexity of the life cycle and the evolution of diversity and disparity in a clade. This simple preliminary analysis suggests a set of topics that deserve priority for scrutiny, including the possible role of saltational evolution in the origination of high diversity and/or disparity, the predictability of morphological evolution following release from a former constraint, and the extent and the possible causes of a positive correlation between diversity and disparity and the complexity of the life cycle.

Morphological, Anatomical, and Physiological Characteristics of Heteroblastic Acacia melanoxylon Grown under Weak Light.

Acacia melanoxylon is a fast-growing macrophanerophyte with strong adaptability whose leaf enables heteromorphic development. Light is one of the essential environmental factors that induces the development of the heteroblastic leaf of A. melanoxylon, but its mechanism is unclear. In this study, the seedlings of A. melanoxylon clones were treated with weak light (shading net with 40% of regular light transmittance) and normal light (control) conditions for 90 d and a follow-up observation. The results show that the seedlings' growth and biomass accumulation were inhibited under weak light. After 60 days of treatment, phyllodes were raised under the control condition while the remaining compound was raised under weak light. The balance of root, stem, and leaf biomass changed to 15:11:74 under weak light, while it was 40:15:45 under control conditions. After comparing the anatomical structures of the compound leaves and phyllode, they were shown to have their own strategies for staying hydrated, while phyllodes were more able to control water loss and adapt to intense light. The compound leaves exhibited elevated levels of K, Cu, Ca, and Mg, increased antioxidant enzyme activity and proline content, and higher concentrations of chlorophyll a, carotenoids, ABA, CTK, and GA. However, they displayed a relatively limited photosynthetic capacity. Phyllodes exhibited higher levels of Fe, cellulose, lignin, IAA content, and high photosynthetic capacity with a higher maximum net photosynthetic rate, light compensation point, dark respiration rate, and water use efficiency. The comparative analysis of compound leaves and phyllodes provides a basis for understanding the diverse survival strategies that heteroblastic plants employ to adapt to environmental changes.

Evolutionary origins of Fibonacci phyllotaxis in land plants.

Fibonacci phyllotaxis is commonly seen in all major groups of land plants. While a precise correlation is found between the internal pattern of the primary vascular system and the external pattern of appendages on the stem surface, it remains a big question how this regularity of Fibonacci phyllotaxis came into being in the course of evolution. Here I address this problem with a model describing phylogenetic and ontogenetic changes in vascular phyllotaxis based on two hypotheses. The first is that the Fibonacci pattern of vascular connection is uniquely determined by the primary arrangement of incipient primordia, the sources of the primary signal system in vascular tissue differentiation. The second is that the surface-area-to-volume ratio of primary vascular tissues serves as a measure of fitness in evolution. The model explains the empirical rule on the manner in which vascular connection is reconfigured during ontogeny, especially during juvenile development. Fossil and phylogenetic evidence suggests that Fibonacci phyllotaxis appeared shortly after the innovation of indefinite lateral organ initiation in a regular sequence.

Age-associated growth control modifies leaf proximodistal symmetry and enabled leaf shape diversification.

Biological shape diversity is often manifested in modulation of organ symmetry and modification of the patterned elaboration of repeated shape elements.1,2,3,4,5 Whether and how these two aspects of shape determination are coordinately regulated is unclear.5,6,7 Plant leaves provide an attractive system to investigate this problem, because they often show asymmetries along the proximodistal (PD) axis of their blades, along which they can also produce repeated marginal outgrowths such as serrations or leaflets.1 One aspect of leaf shape diversity is heteroblasty, where the leaf form in a single genotype is modified with progressive plant age.8,9,10,11 In Arabidopsis thaliana, a plant with simple leaves, SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 9 (SPL9) controls heteroblasty by activating CyclinD3 expression, thereby sustaining proliferative growth and retarding differentiation in adult leaves.12,13 However, the precise significance of SPL9 action for leaf symmetry and marginal patterning is unknown. By combining genetics, quantitative shape analyses, and time-lapse imaging, we show that PD symmetry of the leaf blade in A. thaliana decreases in response to an age-dependent SPL9 expression gradient, and that SPL9 action coordinately regulates the distribution and shape of marginal serrations and overall leaf form. Using comparative analyses, we demonstrate that heteroblastic growth reprogramming in Cardamine hirsuta, a complex-leafed relative of A. thaliana, also involves prolonging the duration of cell proliferation and delaying differentiation. We further provide evidence that SPL9 enables species-specific action of homeobox genes that promote leaf complexity. In conclusion, we identified an age-dependent layer of organ PD growth regulation that modulates leaf symmetry and has enabled leaf shape diversification.