Approaches to the identification of angiosperm leaf remains

During the past 125 years the history of early angiosperms, interpreted through the fossil leaf record has been largely an exercise in paleofloristic studies, ignoring evolution. Imprecise identifications of ancient leaves “matched” to extant genera and families have been used as the basis for reconstructions of paleocommunities and paleoclimates. However, as the result of careful morphological studies of leaf form, venation and cuticular features new insights into the evolution of angiosperms are now available. In this paper considerations are given to the usefulness and shortcomings of leaf form, venation and cuticular analysis as diagnostic tools of plant identification. Many techniques for the study of the morphology of modern and fossil leaves are included in this paper as well as tables outlining features of leaf venation and the epidermis. Careful morphological studies of leaf form (such as the venation and epidermal characters emphasized in this paper) will provide better understanding of the relationships of living angiosperms and transform the fossil leaf record into useful data that can be used to study the evolution of the angiosperms.     

LIRIODENDRON (MAGNOLIACEAE) FROM THE MIOCENE CLARKIA FLORA OF IDAHO

Miocene Liriodendron carpels, whole fruiting structures and leaves from Clarkia and Oviatt Creek sites in northern Idaho are preserved as imprints and compressed fossils in soft lacustrine clays. The isolated carpels are indistinguishable from those described as L. hesperia Berry from the Spokane Latah flora. Fruit aggregates from the type Clarkia and Oviatt Creek localities and leaves from three Clarkia sites are considered to be within the range of variation of the single species L. hesperia. Comparisons were made regarding leaf architecture, lower leaf epidermal structures, leaf flavonoid and steroid analysis, morphological features of receptacles and carpels, and the venation pattern of carpels of the fossil material to the two extant species, L. tulipifera L. (native to southeastern United States) and L. chinense Sarg. (native to southeastern Asia). Leaf architecture features analyzed by standard statistical and canonical tests and marginal venation patterns near the base of leaves suggest that L. hesperia is more similar to L. tulipifera, whereas the size dimensions of lower epidermal cells and the common presence of two sterane compounds imply that L. hesperia is more similar to L. chinense. The fossil species, however, is a distinct taxon indicated by statistical discriminant and canonical tests, leaf base shape, often smaller epidermal cell dimensions, and the shape of round receptacle carpel scars. Both the fossil and the two living Liriodendron species are associated with comparable mixed mesophytic floras.     

CLASSIFICATION OF THE ARCHITECTURE OF DICOTYLEDONOUS LEAVES

A classification of the architectural features of dicot leaves—i.e., the placement and form of those elements constituting the outward expression of leaf structure, including shape, marginal configuration, venation, and gland position—has been developed as the result of an extensive survey of both living and fossil leaves. This system partially incorporates modifications of two earlier classifications: that of Turrill for leaf shape and that of Von Ettingshausen for venation pattern. After categorization of such features as shape of the whole leaf and of the apex and base, leaves are separated into a number of classes depending on the course of their principal venation. Identification of order of venation, which is fundamental to the application of the classification, is determined by size of a vein at its point of origin and to a lesser extent by its behavior in relation to that of other orders. The classification concludes by describing features of the areoles, i.e., the smallest areas of leaf tissue surrounded by veins which form a contiguous field over most of the leaf. Because most taxa of dicots possess consistent patterns of leaf architecture, this rigorous method of describing the features of leaves is of immediate usefulness in both modern and fossil taxonomic studies. In addition, as a result of this method, it is anticipated that leaves will play an increasingly important part in phylogenetic and ecological studies.     

Scutifolium jordanicum gen. et sp. nov. (Cabombaceae), an aquatic fossil plant from the Lower Cretaceous of Jordan, and the relationships of related leaf fossils to living genera

A new species of aquatic plant, Scutifolium jordanicum gen. et sp. nov., Taylor, Brenner & Basha, is described from the Albian of Jordan. The leaves are microphyllous with a symmetrical, elliptical to suborbiculate shape, convex to rounded apex and base, and actinodromous to palinactinodromous primary venation. The peltate, centrally attached petioles are narrow, elongate, and alternately arranged on similarly sized stems. The leaves appear to be thick and have aerenchyma. Comparisons to plants with centrally peltate leaves and palmate venation and to aquatic plants with floating leaves suggest that S. jordanicum belongs to the Cabombaceae lineage within the Nymphaeales. Cladistic analysis including the fossil and living members of the Nymphaeales shows that the S. jordanicum is basal to the living members of the family and has unique characters not found in any living genera. This is the oldest evidence of the Cabombaceae from the Old World. Inclusion of two other Early Cretaceous peltate leaf fossils in the phylogenetic analysis indicates their affinities to Cabombaceae and that some of the shared derived characters for the living members are progressively acquired in the fossils. These data show the Cabombaceae were widespread in Gondwana and Laurasia by the mid-Cretaceous.     

Marsileaceaephyllum, a new genus for marsileaceous macrofossils: leaf remains from the Early Cretaceous (Albian) of southern Gondwana

Three new taxa from Albian, Early Cretaceous assemblages in Gondwana (Australia and Antarctica) and two previously described fossils from the Late Cretaceous and Eocene of North America are attributable to the heterosporous semi-aquatic fern family Marsileaceae. They are assigned to Marsileaceaephyllum, a morphotaxon erected here for sterile remains (whole plants, and isolated leaves and leaflets) of Marsileaceae. The Gondwanan taxa, Marsileaceaephyllum lobatum and Marsileaceaephyllum spp. B-C, have either a cruciform leaflet arrangement or dichotomous and anastomosing venation characteristic of modern Marsileaceae. Two previously established taxa, Marsilea johnhallii and Marsilea sp., which represent sterile Marsileaceae, are also transferred to the new genus (now Marsileaceaephyllum johnhallii and Marsileaceaephyllum sp. A, respectively). Examination of all fossil venation patterns reveals four new venation types not present in extant taxa, suggesting that most fossil Marsileaceae (leaves) are distinct from extant genera, and are likely members of extinct lineages. This is further supported by the absence of modern megaspore types in the Early Cretaceous.     

Phylogenetic relationships of Mangifera species revealed by ITS sequences of nuclear ribosomal DNA and a possibility of their hybrid origin

 The phylogenetic relationships among 14 Mangifera L. species of Thailand were analyzed by comparing sequences of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA (nrDNA). Parsimony and neighbor joining (NJ) analyses revealed that the common mango (M. indica L.) was closely related to M. laurina Bl., M. sylvatica Roxb., and M. oblongifolia Hook. f. Mangifera foetida Lour. and M. odorata Griff. were also related to M. indica in both parsimonious and NJ trees, although these two species are classified into a different subgenus (subgenus Limus) from the subgenus Mangifera to which M. indica belongs. ITS sequence analysis revealed that several species have nucleotide additivity (two different nucleotides at the same locus) at several sites in the ITS region. Also, M. indica had several polymorphisms among cultivars. This finding may suggest a possibility of hybrid origin of Mangifera species, although Mangifera species are all assumed to be diploid having chromosome number of 2n=2x=40. Received February 7, 2001 Accepted October 28, 2001     

Decline of leaf hydraulic conductance with dehydration: relationship to leaf size and venation architecture

Across plant species, leaves vary enormously in their size and their venation architecture, of which one major function is to replace water lost to transpiration. The leaf hydraulic conductance (K(leaf)) represents the capacity of the transport system to deliver water, allowing stomata to remain open for photosynthesis. Previous studies showed that K(leaf) relates to vein density (vein length per area). Additionally, venation architecture determines the sensitivity of K(leaf) to damage; severing the midrib caused K(leaf) and gas exchange to decline, with lesser impacts in leaves with higher major vein density that provided more numerous water flow pathways around the damaged vein. Because xylem embolism during dehydration also reduces K(leaf), we hypothesized that higher major vein density would also reduce hydraulic vulnerability. Smaller leaves, which generally have higher major vein density, would thus have lower hydraulic vulnerability. Tests using simulations with a spatially explicit model confirmed that smaller leaves with higher major vein density were more tolerant of major vein embolism. Additionally, for 10 species ranging strongly in drought tolerance, hydraulic vulnerability, determined as the leaf water potential at 50% and 80% loss of K(leaf), was lower with greater major vein density and smaller leaf size (|r| = 0.85-0.90; P < 0.01). These relationships were independent of other aspects of physiological and morphological drought tolerance. These findings point to a new functional role of venation architecture and small leaf size in drought tolerance, potentially contributing to well-known biogeographic trends in leaf size.     

INVESTIGATIONS OF ANGIOSPERMS FROM THE EOCENE OF NORTH AMERICA: STIPULATE LEAVES OF THE RUBIACEAE INCLUDING A PROBABLE POLYPLOID POPULATION

A comparative study was made of the gross morphology, fine venation and cuticular features of Leitneria fioridana Chapman, the single living representative of the order Leitneriales and Leitneria eocenica (Berry) Brown, presumbaly a related fossil species. In addition to the type material, newly collected fossil specimens were investigated from clay pits in the Middle Eocene, Claiborne Formation, of western Tennessee and Kentucky. Foliate stipules attached to the petioles of several specimens suggest the assignment of this fossil leaf type to the genus Leitneria is incorrect. The nature of the gross morphology, fine venation and cuticular features confirms the misidentification. Previously, various specimens of this fossil leaf type have been placed in eight species of seven genera in seven families of six angiosperm orders, none of which are correct systematically. The gross morphology, venation and cuticular characters of the fossil leaf are distributed among a few extant South American genera of arborescent Rubiaceae. The fossil is an extinct rubiaceous leaf type which cannot be placed within a single modern subfamily, tribe or genus of the family. The organ genus, Paleorubiaceophyllum is proposed for these leaves. Three varieties of a single fossil species, P. eocenicum, are recognized. One variety with epidermal cells nearly twice the size of the others may represent a polyploid population.     

Ectopic divisions in vascular and ground tissues of Arabidopsis thaliana result in distinct leaf venation defects

Leaf venation patterns vary considerably between species and between leaves within a species. A mechanism based on canalization of auxin transport has been suggested as the means by which plastic yet organized venation patterns are generated. This study assessed the plasticity of Arabidopsis thaliana leaf venation in response to ectopic ground or procambial cell divisions and auxin transport inhibition (ATI). Ectopic ground cell divisions resulted in vascular fragments between major veins, whereas ectopic procambial cell divisions resulted in additional, abnormal vessels along major veins, with more severely perturbed lines forming incomplete secondary and higher-order venation. These responses imply limited vascular plasticity in response to unscheduled cell divisions. Surprisingly, a combination of ectopic ground cell divisions and ATI resulted in massive vascular overgrowth. It is hypothesized that the vascular overproduction in auxin transport-inhibited wild-type leaves is limited by simultaneous differentiation of ground cells into mesophyll cells. Ectopic ground cell divisions may negate this effect by providing undifferentiated ground cells that respond to accumulated auxin by differentiation into vascular cells.     

Leaves of Cercocarpus mixteca n. sp. (Rosaceae) from Oligocene sediments, near Tepexi de Rodríguez, Puebla

A new species of Cercocarpus, Cercocarpus mixteca Velasco de León & Cevallos-Ferriz, is described based on leaf impressions from the Los Ahuehuetes locality, near Tepexi de Rodríguez, Puebla, Mexico. The lamina is obovate, 1.3cm in length by 0.5cm in width, has a serrate margin in its distal fourth, craspedodromous venation with a single straight mid-vein and two to four pairs of secondary ones, and areols that tend to be quadrangular in shape. A phenetic analysis of the agglomerative, non-hierarchical type, with mean linkage, is applied using 22 OTUs and 34 character states. The morphological characters observed on the leaves of the new fossil plants support the recognition of a new taxon closely related to the extant Cercocarpus paucidentatus growing naturally in northern Mexico. Its microphyll size corresponds with the temperate to xeric climate postulated for the Los Ahuehuetes locality; this further suggests that some taxa, like Cercocarpus, have a long history in low latitude North America. In this particular case, the extant Cercocarpus fothergilloides and Cercocarpus macrophyllus could, as they were able to colonise new humid and xeric areas, represent descendants of C. mixteca.     

INVESTIGATIONS OF ANGIOSPERMS FROM THE EOCENE OF SOUTHEASTERN NORTH AMERICA: PHILODENDRON LEAF REMAINS

Large leaves, new to the fossil record, from the Claiborne Formation in western Tennessee have been collected and analyzed. Careful analyses of venation and cuticular anatomy indicate that these fossil leaves contain specific characters found in modern species of the genus Philodendron subgenus Meconostigma. Features of venation were taxonomically more useful than other features. Reports of fossils of the Araceae are scarce and those few fossils assigned to the family frequently have been inadequately studied. This report establishes a reliable and carefully documented occurrence of the Araceae in the fossil record, and provides information concerning the differentiation of the family in time and the distributions of the genus Philodendron.     

Studies on the leaf anatomy of Euphorbia: II. Venation patterns*

Leaf venation patterns of 150 species of Euphorbia are presented and their value as a diagnostic feature in the genus assessed. Based on the gross venation patterns, the species have been grouped into uni-, bi-, tri-veined and special categories. The majority of the species studied belonged to the tri-veined category, in which ornamentation of the veins and the course of traces in the lamina proved useful additional characters. Features such as the size of the areoles, the number of vein endings, and their further ramifications and composition in each areole varied in the same leaf or in different leaves of a given species. Furthermore, no direct correlation could be established between the areole size and the numbers of vein endings and tips per areole. Forty species possess a parenchymatous vein sheath. Globular chloroplasts showed up conspicuously in the sheath cells of a few species. Dilated tracheidal elements, localized on the venules or at the tips of vein endings, are characteristic of the xerophytic species. In some instances, correlation was apparent between plants grouped according to their venation patterns and their habit.     

Venation networks and the origin of the leaf economics spectrum

The leaf economics spectrum describes biome-invariant scaling functions for leaf functional traits that relate to global primary productivity and nutrient cycling. Here, we develop a comprehensive framework for the origin of this leaf economics spectrum based on venation-mediated economic strategies. We define a standardized set of traits - density, distance and loopiness - that provides a common language for the study of venation. We develop a novel quantitative model that uses these venation traits to model leaf-level physiology, and show that selection to optimize the venation network predicts the mean global trait-trait scaling relationships across 2548 species. Furthermore, using empirical venation data for 25 plant species, we test our model by predicting four key leaf functional traits related to leaf economics: net carbon assimilation rate, life span, leaf mass per area ratio and nitrogen content. Together, these results indicate that selection on venation geometry is a fundamental basis for understanding the diversity of leaf form and function, and the carbon balance of leaves. The model and associated predictions have broad implications for integrating venation network geometry with pattern and process in ecophysiology, ecology and palaeobotany.     

MUSOPSIS N. GEN.: A BANANA-LIKE LEAF GENUS FROM THE EARLY TERTIARY OF EASTERN NORTH GREENLAND

A fossil flora from the Late Paleocene-Early Eocene Thyra Ø Formation of eastern North Greenland (paleolatitude 77° N) has yielded monocotyledon leaf impressions with characters seen only in the closely related modem species in the families of Heliconiaceae, Musaceae, and Strelitziaceae. The combination of large costae widths and parallel, nonanastomosing, lateral veins that depart at right angles from the costae in the fossil material are features present only in leaves of extant species from these families. Three basic venation patterns also are recognized in the modem species of these families, but except for the genera Strelitzia and Phenakospermum, none of these patterns are present exclusively in any one family. Musopsis n. gen. is created for the fossil material from Greenland, but it is considered a form genus due to the lack of gross morphological features that can be used for separating leaves of the modem genera in Heliconiaceae, Musaceae, and Strelitiziaceae. It is the first known Arctic occurrence of fossil leaf material resembling this modem group of taxa.     

Foliar architecture of vanilloid orchids: insights into the evolution of reticulate leaf venation in monocotyledons

Leaves of representative taxa within the vanilloid clade of Orchidaceae were cleared and their venation patterns studied. Within subtribe Pogoniinae, Isotria and Pogonia exhibit a prominent reticulate venation pattern, although only Pogonia ophioglossoides is characterized by free vein endings. Within subtribe Vanillinae, all species of Epistephium , as well as the New Caledonian endemics, Eriaxis rigida and Clematepistephium smilacifolium , have reticulate-veined leaves that are characterized by numerous free vein endings. Leaves of most species of Vanilla exhibit a parallel-veined pattern stereotypical of monocots, although branched secondaries with free endings were observed in V. africana. Most members of subtribe Galeolinae are 'saprophytes' with reduced bract-like leaves. Vascular bundles enter these leaf-homologues in a parallel manner, but quickly ram+ throughout the tissue in an irregular manner. Leaf venation is used to hypothesize patterns of generic relationships within the vanilloid clade. Molecular evidence for phylogenetic relationships among angiosperms indicates that reticulate leaf venation has arisen independently in several unrelated monocot families, including the vanilloid Orchidaceae, perhaps by a similar evolutionary scenario.     

Differential leaf expansion can enable hydraulic acclimation to sun and shade

Although leaf size is one of the most responsive plant traits to environmental change, the functional benefits of large versus small leaves remain unclear. We hypothesized that modification of leaf size within species resulting from differences in irradiance can allow leaves to acclimate to different photosynthetic or evaporative conditions while maintaining an efficient balance between hydraulic supply (vein density) and evaporative demand. To test this, we compared the function and anatomy of leaf hydraulic systems in the leaves of a woody angiosperm (Toona ciliata M. Roem.) grown under high and low irradiance in controlled conditions. Our results confirm that in this species, differential leaf expansion regulates the density of veins and stomata such that leaf hydraulic conductance and stomatal conductance remain proportional. A broader sample of field-grown tree species suggested that differences in leaf venation and stomatal traits induced by sun and shade were not regulated by leaf size in all cases. Our results, however, suggest that leaf size plasticity can provide an efficient way for plants to acclimate hydraulic and stomatal conductances to the contrasting evaporative conditions of sun and shade.     

A morpho-anatomical characterisation of Myrothamnus moschatus (Myrothamnaceae) under the aspect of desiccation tolerance

Morpho-anatomical traits of the rarely studied dicotyledonous desiccation-tolerant shrub Myrothamnus moschatus were examined and compared for the first time to Myrothamnus flabellifolius under the aspect of desiccation tolerance. Both species almost exclusively occur on rock outcrops and differ mainly in their geographic range and leaf morphology (fan-shaped in M. flabellifolius, lanceolate in M. moschatus) but have a very similar leaf and wood anatomy, except for the lack of hydathodes in M. moschatus. Both species adopt the parallel leaf venation of monocots, although this is more pronounced in M. moschatus. This provides a mechanical and protective advantage over the net venation pattern of most dicots and facilitates the reversible, drought-induced, accordion-like leaf contraction. The sclerenchyma, as a stabilising tissue, is mainly confined to vascular bundles in leaves of both species. Here, mechanical support seems to be less crucial for survival in long periods of drought than other morpho-anatomical traits (e.g. parallel leaf venation).     

Water stress-induced modifications of leaf hydraulic architecture in sunflower: co-ordination with gas exchange

The hydraulic architecture, water relationships, and gas exchange of leaves of sunflower plants, grown under different levels of water stress, were measured. Plants were either irrigated with tap water (controls) or with PEG600 solutions with osmotic potential of -0.4 and -0.8 MPa (PEG04 and PEG08 plants, respectively). Mature leaves were measured for hydraulic resistance (R(leaf)) before and after making several cuts across minor veins, thus getting the hydraulic resistance of the venation system (R(venation)). R(leaf) was nearly the same in controls and PEG04 plants but it was reduced by about 30% in PEG08 plants. On the contrary, R(venation) was lowest in controls and increased in PEG04 and PEG08 plants as a likely result of reduction in the diameter of the veins' conduits. As a consequence, the contribution of R(venation) to the overall R(leaf) markedly increased from controls to PEG08 plants. Leaf conductance to water vapour (g(L)) was highest in controls and significantly lower in PEG04 and PEG08 plants. Moreover, g(L) was correlated to R(venation) and to leaf water potential (psi(leaf)) with highly significant linear relationships. It is concluded that water stress has an important effect on the hydraulic construction of leaves. This, in turn, might prove to be a crucial factor in plant-water relationships and gas exchange under water stress conditions.     

Palaeogene leaf compressions of myrtaceous affinity from Pasekovo, Middle Russian Upland, southern European Russia

Leaf compressions from Pasckovo, Voronezh oblast, a Late Eocene site, constitute the first record of mummified 'paper coal' leaves of myrtaceous affinity in Russia. The fossil leaves are assigned to the organ-genus Rhodomyrtophyllum Rüffle & Jähnichen (Myrtaceac) on the basis of epidermal and venation features. The exceptionally high level of preservation is illustratcd by LM and SEM reproductions of anomo-, anomostauro- and laterocytic stomatal sets including giant stomata, double cuticular rims, trichome bases (scars) and a buttressed structure of the epidermal cell wall. Leaf structure is mesoxeromorphic. In both western and eastern Europe, species of the genus do not post-date the terminal Eocene and are the index taxa for the middle to late Eocene interval. The newly described Russian species R.pasekovicum Vikulin, sp.now is considered against the background of British, German and Czech extinct species along with comparisons with possible 'nearest living relatives' which include mesoxeromorphic Eugenia L., Syzygium Gartn.and Rhodomyrtus DC, from South-East Asia, Australia and South America. Ancient Tethys-ocean migrational channels may explain European palacofloristic relations with the Australo-Malesian area during Eocene timeS. A later Eocene age is postulated of the Pasekovo palaeoflora in contrast to the early Oligocene age suggested by other authors.     

Mitrephora sirikitiae (Annonaceae): a remarkable new species endemic to northern Thailand

A new species of Annonaceae, Mitrephora sirikitiae , is described from Mae Hong Son Province in northern Thailand. It is easily distinguished from the seven species of Mitrephora previously recorded from Thailand due to its very large, showy flowers. It is most similar to M. winitii , but differs in its larger flowers, with inner petals that become undulate at maturity. The profuse blooming of the new species and its large flowers with mild fragrance suggest that may be of significant horticultural potential.