Compound-leafed Species Research Articles

Rewiring of a KNOXI regulatory network mediated by UFO underlies the compound leaf development in Medicago truncatula

Class I KNOTTED-like homeobox (KNOXI) genes are parts of the regulatory network that control the evolutionary diversification of leaf morphology. Their specific spatiotemporal expression patterns in developing leaves correlate with the degrees of leaf complexity between simple-leafed and compound-leafed species. However, KNOXI genes are not involved in compound leaf formation in several legume species. Here, we identify a pathway for dual repression of MtKNOXI function in Medicago truncatula. PINNATE-LIKE PENTAFOLIATA1 (PINNA1) represses the expression of MtKNOXI, while PINNA1 interacts with MtKNOXI and sequesters it to the cytoplasm. Further investigations reveal that UNUSUAL FLORAL ORGANS (MtUFO) is the direct target of MtKNOXI, and mediates the transition from trifoliate to pinnate-like pentafoliate leaves. These data suggest a new layer of regulation for morphological diversity in compound-leafed species, in which the conserved regulators of floral development, MtUFO, and leaf development, MtKNOXI, are involved in variation of pinnate-like compound leaves in M. truncatula.

Silencing of PhLA, a CIN-TCP gene, causes defected petal conical epidermal cell formation and results in reflexed corolla lobes in petunia

BackgroundTCP-domain proteins, plant specific transcription factors, play important roles in various developmental processes. CIN-TCPs control leaf curvature in simple leaf species while regulate leaf complexity in compound leaf species. However, the knowledge was largely based on findings in few model species. To extend our knowledge on this group of proteins in Solanaceae species, we identified a CIN-TCP gene from petunia, and studied its functions using virus-induced gene silencing (VIGS).ResultsConsistently, silencing of CIN-TCPs increases complexity of tomato leaves, and enhances leaf curvature in Nicotiana benthamiana. However, in petunia (Petunia hybrida), silencing of petunia LA, a CIN-TCP, through VIGS did not obviously affect leaf shape. The silencing, however, enhanced petal curvature. The event was associated with petal expansion at the distal portion where epidermal cell size along the midribs was also increased. The enlarged epidermal cells became flattened. Although shapes of PhLA-silenced flowers largely resemble phmyb1 mutant phenotype, PhMYB1 expression was not affected when PhLA was specifically silenced. Therefore, both PhLA and PhMYB1 are required to regulate flower morphology. In corolla, PhLA and miR319 deferentially express in different regions with strong expressions in limb and tube region respectively.ConclusionsIn conclusion, unlike LA-like genes in tomato and N. benthamiana, PhLA plays a more defined role in flower morphogenesis, including petal curvature and epidermal cell differentiation.

CLAUSA Is a MYB Transcription Factor That Promotes Leaf Differentiation by Attenuating Cytokinin Signaling.

Leaf morphogenesis and differentiation are highly flexible processes, resulting in a large diversity of leaf forms. The development of compound leaves involves an extended morphogenesis stage compared with that of simple leaves, and the tomato (Solanum lycopersicum) mutant clausa (clau) exposes a potential for extended morphogenesis in tomato leaves. Here, we report that the CLAU gene encodes a MYB transcription factor that has evolved a unique role in compound-leaf species to promote an exit from the morphogenetic phase of tomato leaf development. We show that CLAU attenuates cytokinin signaling, and that clau plants have increased cytokinin sensitivity. The results suggest that flexible leaf patterning involves a coordinated interplay between transcription factors and hormones.

Ectopic expression of the PttKN1 gene in Cardamine hirsuta mediated via the floral dip method

PttKN1 gene (Populus tremula × P. tremuloides KNOTTED1) was isolated from the vascular cambium of a hybrid aspen. Previous studies on transformed plants with the PttKN1 gene suggested that it plays roles in plant development (typically in meristem initiation), maintenance and organogenesis in simple-leaved species. To investigate the gene functions further, sequence analysis of the deduced amino acid was conducted. The results suggested that the gene belongs to the class I KNOX gene (KNOTTED1-like homeobox genes) family and might play important roles in plant development by coding a transcription factor. The gene was introduced into Cardamine hirsuta using the floral dip method mediated via Agrobacterium tumefaciens. The primary transformed plants were obtained via kanamycin selection. Compared to the wild type, the kanamycin resistant plants demonstrated several morphological alterations, such as abnormal cotyledons, abnormal shoot meristem, flattened stem, and lobed and cup-shaped leaves. RT-PCR results showed that the above five types of kanamycin resistant plants expressed the same specific PttKN1 gene band. This suggested that the morphological alterations were caused by the insertion and expression of the gene. However, these phenotypes were similar to other PttKN1 transformed plants, despite the fact that C. hirsuta is a species with compound leaves and the other species have simple leaves. Therefore, the functions of the PttKN1 gene on compound-leaf species have yet to be investigated via the comparison between related species such as Arabidopsis thaliana and C. hirsuta.

Assessing the potential for leaf-off LiDAR data to model canopy closure in temperate deciduous forests

Estimates of canopy closure have many important uses in forest management and ecological research. Field measurements, however, are typically not practical to acquire over expansive areas or for large numbers of locations. This problem has been addressed, in recent years, through the use of airborne light detection and ranging (LiDAR) technology which has proven effective in modeling canopy closure remotely. The techniques developed to use LiDAR for this purpose have been designed and evaluated for datasets acquired during leaf-on conditions. However, a large number of LiDAR datasets are acquired during leaf-off conditions since their primary purpose is to generate bare-earth Digital Elevation Models. In this paper, we develop and evaluate techniques for leveraging small-footprint leaf-off LiDAR data to model leaf-on canopy closure in temperate deciduous forests.We evaluate three techniques for modeling canopy closure: (1) the canopy-to-total-return-ratio (CTRR), (2) the canopy-to-total-pixel-ratio (CTPR), and (3) the hemispherical-viewshed (HV). The first technique has been used widely, in various forms, and has been shown to be effective with leaf-on LiDAR datasets. The CTRR technique that we tested uses the first-return LiDAR data only. The latter two techniques are new contributions that we develop and present in this paper. These techniques use Canopy Height Models (CHM) to detect significant gaps in the forest canopy which are of primary importance in estimating closure.The techniques we tested each showed good promise for predicting canopy closure using leaf-off LiDAR data with the CTPR and HV models having particularly high correlations with closure estimates from hemispherical photographs. The CTRR model had performance on par with results from previous studies that used leaf-on LiDAR, although, with leaf-off data the model tended to be negatively biased with respect to species having simple and compound leaf types and positively biased for coniferous species. The CTPR and HV models also showed some slight negative biases for compound-leaf species. The biases for the CTPR and HV models were mitigated when the CHM data were smoothed to fill in small gaps. The CHM-based models were robust to changes in the CHM model resolution which suggests that these methods may be applicable to a variety of small-footprint LiDAR datasets. In this research, the new CTPR and HV methods showed a strong ability to predict canopy closure using leaf-off data, however, future work will be needed to test the applicability of the models to variations in LiDAR datasets, forest types, and topography.

Regulation of Compound Leaf Development by PHANTASTICA in Medicago truncatula

Plant leaves, simple or compound, initiate as peg-like structures from the peripheral zone of the shoot apical meristem, which requires class I KNOTTED-LIKE HOMEOBOXI (KNOXI) transcription factors to maintain its activity. The MYB domain protein encoded by the ASYMMETRIC LEAVES1/ROUGH SHEATH2/PHANTASTICA (ARP) gene, together with other factors, excludes KNOXI gene expression from incipient leaf primordia to initiate leaves and specify leaf adaxial identity. However, the regulatory relationship between ARP and KNOXI is more complex in compound-leafed species. Here, we investigated the role of ARP and KNOXI genes in compound leaf development in Medicago truncatula. We show that the M. truncatula phantastica mutant exhibited severe compound leaf defects, including curling and deep serration of leaf margins, shortened petioles, increased rachises, petioles acquiring motor organ characteristics, and ectopic development of petiolules. On the other hand, the M. truncatula brevipedicellus mutant did not exhibit visible compound leaf defects. Our analyses show that the altered petiole development requires ectopic expression of ELONGATED PETIOLULE1, which encodes a lateral organ boundary domain protein, and that the distal margin serration requires the auxin efflux protein M. truncatula PIN-FORMED10 in the M. truncatula phantastica mutant.

Development of leaf shape in two North American native species ofAmpelopsis(Vitaceae)

The Vitaceae (grape family) is a large group of plants with a world-wide distribution. Despite the economic importance of cultivated grapes (Vitis vinifera L.), very little is known about leaf development in the Vitaceae. The genus Ampelopsis contains simple- and compound-leafed forms making it an appropriate genus in which to conduct a comparative developmental study. The Ampelopsis clade diverged early from the rest of the Vitaceae, and the simple- and compound-leaf morphologies are separated into two distinct lineages, leaving the question of how this morphological disjunct arose and what the ancestral leaf shape of this genus might have been. Thus, understanding the development of this trait will clarify our understanding of basic Vitaceae leaf characters and of leaf shape evolution. We found that although the primordia of both the simple- and compound-leafed species shared a fundamental plan that would be predicted to result in a mature complex leaf shape at the outset, their developmental trajectory diverges early in development. We also identified key morphological landmarks that suggest homology between the two species. Based on current phylogenetic analysis, we discuss how our developmental study likely points to homologies that reflect their recent common ancestry despite the morphological disjunct observed in the genus.

TALE and Shape: How to Make a Leaf Different.

The Three Amino acid Loop Extension (TALE) proteins constitute an ancestral superclass of homeodomain transcription factors conserved in animals, plants and fungi. In plants they comprise two classes, KNOTTED1-LIKE homeobox (KNOX) and BEL1-like homeobox (BLH or BELL, hereafter referred to as BLH), which are involved in shoot apical meristem (SAM) function, as well as in the determination and morphological development of leaves, stems and inflorescences. Selective protein-protein interactions between KNOXs and BLHs affect heterodimer subcellular localization and target affinity. KNOXs exert their roles by maintaining a proper balance between undifferentiated and differentiated cell state through the modulation of multiple hormonal pathways. A pivotal function of KNOX in evolutionary diversification of leaf morphology has been assessed. In the SAM of both simple- and compound-leafed seed species, downregulation of most class 1 KNOX (KNOX1) genes marks the sites of leaf primordia initiation. However, KNOX1 expression is re-established during leaf primordia development of compound-leafed species to maintain transient indeterminacy and morphogenetic activity at the leaf margins. Despite the increasing knowledge available about KNOX1 protein function in plant development, a comprehensive view on their downstream effectors remains elusive. This review highlights the role of TALE proteins in leaf initiation and morphological plasticity with a focus on recent advances in the identification of downstream target genes and pathways.

Multiple Components are Integrated to Determine Leaf Complexity in Lotus japonicus

Transcription factors and phytohormones have been reported to play crucial roles to regulate leaf complexity among plant species. Using the compound-leafed species Lotus japonicus, a model legume plant with five visible leaflets, we characterized four independent mutants with reduced leaf complexity, proliferating floral meristem (pfm), proliferating floral organ-2 (pfo-2), fused leaflets1 (ful1) and umbrella leaflets (uml), which were further identified as loss-of-function mutants of Arabidopsis orthologs LEAFY (LFY), UNUSUAL FLORAL ORGANS (UFO), CUP-SHAPED COTYLEDON 2 (CUC2) and PIN-FORMED 1 (PIN1), respectively. Comparing the leaf development of wild-type and mutants by a scanning electron microscopy approach, leaflet initiation and/or dissection were found to be affected in these mutants. Expression and phenotype analysis indicated that PFM/LjLFY and PFO/LjUFO determined the basipetal leaflet initiation manner in L. japonicus. Genetic analysis of ful1 and uml mutants and their double mutants revealed that the CUC2-like gene and auxin pathway also participated in leaflet dissection in L. japonicus, and their functions might influence cytokinin biogenesis directly or indirectly. Our results here suggest that multiple genes were interplayed and played conserved functions in controlling leaf complexity during compound leaf development in L. japonicus.

TheREDUCED LEAFLETGenes Encode Key Components of the trans-Acting Small Interfering RNA Pathway and Regulate Compound Leaf and Flower Development inLotus japonicus

The endogenous trans-acting small interfering RNA (ta-siRNA) pathway plays a conserved role in adaxial-abaxial patterning of lateral organs in simple-leafed plant species. However, its function in compound-leafed species is largely unknown. Using the compound-leafed species Lotus japonicus, we identified and characterized two independent mutants, reduced leaflet1 (rel1) and rel3, whose most conspicuous defects in compound leaves are abaxialized leaflets and reduction in leaflet number. Concurrent mutations in REL genes also compromise flower development and result in radial symmetric floral organs. Positional cloning revealed that REL1 and REL3 encode the homologs of Arabidopsis (Arabidopsis thaliana) SUPPRESSOR OF GENE SILENCING3 and ARGONAUTE7/ZIPPY, respectively, which are key components of the ta-siRNA pathway. These observations, together with the expression and functional data, demonstrated that the ta-siRNA pathway plays conserved yet distinct roles in the control of compound leaf and flower development in L. japonicus. Moreover, the phenotypic alterations of lateral organs in ta-siRNA-deficient mutants and the regulation of downstream targets by the ta-siRNA pathway in L. japonicus were similar to those in the monocots but different from Arabidopsis, indicating many parallels between L. japonicus and the monocots in the control of lateral organ development by the ta-siRNA pathway.

Coordination of leaf development via regulation of KNOX1 genes

Class I KNOTTED1-LIKE HOMEOBOX (KNOX1) genes are expressed in the shoot apical meristem (SAM) to effect its formation and maintenance. KNOX1 genes are also involved in leaf shape control throughout angiosperm evolution. Leaves can be classified as either simple or compound, and KNOX1 expression patterns in leaf primordia are highly correlated with leaf shape; in most simple-leafed species, KNOX1 genes are expressed only in the SAM but not in leaf primordia, while in compound-leafed species they are expressed both in the SAM and leaf primordia. How can KNOX1 expression be maintained to a high degree in the SAM, but simultaneously be so variable in leaves? This dichotomy suggests that the processes of leaf and SAM development have been compartmentalized during evolution. Here, we introduce our findings regarding the regulation of expression of SHOOT MERISTEMLESS, a KNOX1 gene, together with a brief review of KNOX1 genes from an evolutionary viewpoint. We also present our findings regarding another aspect of KNOX1 regulation via a protein-protein interaction network involved in the natural variation in leaf shape. Both aspects of KNOX1 regulation could be utilized for fine-tuning leaf morphology during evolution without affecting the essential function of KNOX genes in the shoot.

The effects of leaf size, leaf habit, and leaf form on leaf/stem relationships in plant twigs of temperate woody species

AbstractQuestion: Do thick‐twigged/large‐leaf species have an advantage in leaf display over their counterparts, and what are the effects of leaf habit and leaf form on the leaf‐stem relationship in plant twigs of temperature broadleaf woody species?Location: Gongga Mountain, southwest China.Methods: (1) We investigated stem cross‐sectional area and stem mass, leaf area and leaf/lamina mass of plant twigs (terminal branches of current‐year shoots) of 89 species belonging to 55 genera in 31 families. (2) Data were analyzed to determine leaf‐stem scaling relationships using both the Model type II regression method and the phylogenetically independent comparative (PIC) method.Results: (1) Significant, positive allometric relationships were found between twig cross‐sectional area and total leaf area supported by the twig, and between the cross‐sectional area and individual leaf area, suggesting that species with large leaves and thick twigs could support a disproportionately greater leaf area for a given twig cross‐sectional area. (2) However, the scaling relationships between twig stem mass and total leaf area and between stem mass and total lamina mass were approximately isometric, which indicates that the efficiency of deploying leaf area and lamina mass was independent of leaf size and twig size. The results of PIC were consistent with these correlations. (3) The evergreen species were usually smaller in total leaf area for a given twig stem investment in terms of both cross‐sectional area and stem mass, compared to deciduous species. Leaf mass per area (LMA) was negatively associated with the stem efficiency in deploying leaf area. (4) Compound leaf species could usually support a larger leaf area for a given twig stem mass and were usually larger in both leaf size and twig size than simple leaf species.Conclusions: Generally, thick‐twigged/large‐leaf species do not have an advantage over their counterparts in deploying photosynthetic compartments for a given twig stem investment. Leaf habit and leaf form types can modify leaf‐stem scaling relationships, possibly because of contrasting leaf properties. The leaf size‐twig size spectrum is related to the LMA‐leaf life span dimension of plant life history strategies.

Fisionomia e estrutura de uma floresta estacional montana do maciço da Borborema, Pernambuco - Brasil

The physiognomy and structure of the tree component of a montane forest fragment located in an area at 900 m altitude (8o11'144 -8o12'27 S and 36o23'730 -36o24'638 W), with 948 mm of annual rainfall, and deep soil were studied. All alive or dead standing plants with diameter at breast height > 5 cm, within 50 contiguous plots, 10 x 20 m each, had their trunk diameter, total plant height and canopy projection area measured. The species were classified in relation to their leaf size, deciduousness, and lamina composition. A total of 61 species were sampled plus one unidentified taxon. Plant density, basal area and average and maximum height and diameters were 1,553 plant ha-1, 39 m2 ha-1, 10.3 and 30 m, 14.2 and 105 cm, respectively. About 75% of all alive plants had between 1.4 and 3 m high. The distribution of basal and canopy areas according to height class indicated two ranges of plant concentrations: 7-13 and 19-22 m. The forest fragment represents a transition between humid and dry montane forests. Most of the species are evergreen with simple microphyllous leaves (Casearia sylvestris Sw., Guapira nitida (Schmidt) Lundell, Marlierea clausseniana (O. Berg) Kiaersk., Ocotea aff. elegans Mez, Plinia sp.), they occurred in the canopy of the forest, together with notophyllous species (Amaioua cf. guianensis Aubl. and Roupala paulensis Sleumer) and mesophyllous (Fabaceae sp. and Inga marginata Willd.). The emergent trees are mostly deciduous and compound leaf species, such as Copaifera trapezifolia Hayne and Eriotheca crenulaticalyx A. Robyns.

Regulation of SHOOT MERISTEMLESS genes via an upstream-conserved noncoding sequence coordinates leaf development

The indeterminate shoot apical meristem of plants is characterized by the expression of the Class 1 KNOTTED1-LIKE HOMEOBOX (KNOX1) genes. KNOX1 genes have been implicated in the acquisition and/or maintenance of meristematic fate. One of the earliest indicators of a switch in fate from indeterminate meristem to determinate leaf primordium is the down-regulation of KNOX1 genes orthologous to SHOOT MERISTEMLESS (STM) in Arabidopsis (hereafter called STM genes) in the initiating primordia. In simple leafed plants, this down-regulation persists during leaf formation. In compound leafed plants, however, KNOX1 gene expression is reestablished later in the developing primordia, creating an indeterminate environment for leaflet formation. Despite this knowledge, most aspects of how STM gene expression is regulated remain largely unknown. Here, we identify two evolutionarily conserved noncoding sequences within the 5' upstream region of STM genes in both simple and compound leafed species across monocots and dicots. We show that one of these elements is involved in the regulation of the persistent repression and/or the reestablishment of STM expression in the developing leaves but is not involved in the initial down-regulation in the initiating primordia. We also show evidence that this regulation is developmentally significant for leaf formation in the pathway involving ASYMMETRIC LEAVES1/2 (AS1/2) gene expression; these genes are known to function in leaf development. Together, these findings reveal a regulatory point of leaf development mediated through a conserved, noncoding sequence in STM genes.