The shoot apex of Podostemaceae: De novo structure or reduction of the conventional type?

The paper reviews and discusses various interpretations of the shoot apex of Podostemaceae with special reference to subfamily Podostemoideae. Main questions concern (1) the proposed absence of a shoot apical meristem (SAM) in apical “meristemless” shoot tips of Podostemoideae and, as the consequence, the endogenous inception of leaf-borne leaves and branches and (2) the predicted stem bifurcation below a “terminal” dithecous (double-sheathed) leaf positioned instead of a shoot apex, as it is reported for subfamily Podostemoideae. Does the “meristemless” shoot apex represent a true evolutionary novelty? Does the view of stem bifurcation represent a new ramification pattern with the consequence that the “classical root–shoot model” of angiosperms is not valid for Podostemaceae? Both interpretations do not conform to previous studies that are complemented here by new data on the SAM of Zeylanidium olivaceum and Thelethylax minutiflora (Podostemoideae). Although a SAM is difficult to observe in the vegetative shoots of many Podostemoideae, it becomes well visible when the shoot passes into the flowering stage approaching the conspicuous shoot apex of floriferous shoots. The arguments of the absence of a SAM in vegetative shoots are not convincing and the endogenous origin of “leaf-borne leaves” appears questionable. Consequently, the “meristemless” shoot apex cannot be considered as a structure having evolved de novo. In the less advanced subfamilies Tristichoideae and Weddellinoideae, the leaf primordia develop only from a few apical cells of the outer shoot layer. This allows the conclusion that the surface layer of the apex in these subfamilies corresponds to the horizontally spread single-layered apical meristem of subfamily Podostemoideae. Similarly, the view of shoot bifurcation does not conform to the diachsial–sympodial branching pattern occurring in the cymose inflorescences of many Podostemoideae. This fact contradicts the presence of a terminal leaf.     

Developmental morphology of the shoot in Weddellina squamulosa and implications for shoot evolution in the Podostemaceae

BACKGROUND AND AIMS: In angiosperms, the shoot apical meristem produces a shoot system composed of stems, leaves and axillary buds. Podostemoideae, one of three subfamilies of the river-weed family Podostemaceae, have a unique 'shoot' that lacks a shoot apical meristem and is composed only of leaves. Tristichoideae have been interpreted to have a shoot apical meristem, although its branching pattern is uncertain. The shoot developmental pattern in Weddellinoideae has not been investigated with a focus on the meristem. Weddellinoideae are in a phylogenetically key position to reveal the process of shoot evolution in Podostemaceae. METHODS: The shoot development of Weddellina squamulosa, the sole species of Weddellinoideae, was investigated using scanning electron microscopy and semi-thin serial sections. KEY RESULTS: The shoot of W. squamulosa has a tunica-corpus-organized apical meristem. It is determinate and successively initiates a new branch extra-axillarily at the base of an immediately older branch, resulting in a sympodial, approximately plane branching pattern. Large scaly leaves initiate acropetally on the flanks of the apical meristem, as is usual in angiosperms, whereas small scaly leaves scattered on the stem initiate basipetally in association with the elongation of internodes. CONCLUSIONS: Weddellinoideae, like Tristichoideae, have a shoot apical meristem, leading to the hypothesis that the meristem was lost in Podostemoideae. The patterns of leaf formation in Podostemoideae and shoot branching in Weddellinoideae are similar in that these organs arise at the bases of older organs. This similarity leads to another hypothesis that the 'branch' in Weddellinoideae (and possibly Tristichoideae) and the 'leaf' in Podostemoideae are comparable, and that the shoot apical meristem disappeared in the early evolution of Podostemaceae.     

Shoot apex and spathella: two problematical structures of Podostemaceae–Podostemoideae

The presence of a shoot apex and shoot apical meristem (SAM), said to be absent in subfamily Podostemoideae (Podostemaceae), is confirmed for Marathrum utile and M. foeniculaceum. The vegetative shoot axis is terminated by a small group of meristematic cells which are surrounded by the tissue of the adnate bases of foliage leaves. The slightly bulged tip of the shoot apex is embraced by the youngest leaf, facing the apex with its adaxial side. The study also refers to the spathella, a cup-shaped structure covering obligatorily the young flower bud in Podostemoideae. The occurrence of two separate peaks in the young spathella of M. foeniculaceum supports the view that the spathella is formed by two fused bracts (hypsophylls). The two bracts are perpendicular to the distichous foliage leaves below the spathella. The scaly leaflet on the spathella of A. latifolia apparently does not represent a rudimentary blade of the spathella, but is interpreted as a separate bract. The occasional occurrence of scales below or above the spathella points to a reduction of bracts that were originally present in greater number on the pedicels.     

Diamantina lombardii – an odd Brazilian member of the Podostemaceae

This paper complements the diagnosis of the recently described genus Diamantina and its only species Diamantina lombardii Novelo, Philbrick and Irgang from Minas Gerais (Brazil). Four new features not known from other Podostemaceae–Podostemoideae are documented by microtome sections and SEM graphs: (i) The digitate foliage leaves lack vascular tissue completely. (ii) Leafy shoots produce one or two flowers in terminal and subterminal position. The spathella subtending the subterminal flower is scale-like and positionally homologous to a digitate bract (leaf), whereas the spathella covering the terminal flower bud is tubular (as usual for Podostemoideae). (iii) The usually rudimentary androecium consists of two stamens which form a complete whorl together with three inconspicuous tepals around the gynophore. (iv) The bilocular ovary has an apical cleft. Each carpel tip (hollow inside) is topped by a prominent horn-like stigma. Additional peculiar features of D. lombardii (already mentioned in Philbrick et al., 2004. Syst. Bot. 29, 109–117) are shown: presence of a prominent gynophore (mainly known from African Podostemoideae), and digitate leaves (as found in Cladopus from Eastern Asia to NE Australia).     

Microspore development in Podostemaceae-Podostemoideae, with implications on the characterization of the subfamilies

Subfamilies Podostemoideae and Tristichoideae of the aquatic flowering plant family Podostemaceae are conventionally characterized by a different mode of microsporogenesis. Simultaneous meiotic division into the four microspores is found in Tristichoideae, successive meiotic division is said to be typical of Podostemoideae. In contrast, the results of the present study reveal that in subfamily Podostemoideae both modes of microsporogenesis occur. This is exemplified by the early pollen development of two neotropical species: Apinagia latifolia and Marathrum rubrum. Successive versus simultaneous meiotic cytokinesis are thus not differential characters of the two subfamilies. It is worthy to note that successive cytokinesis occurs in a family (Podostemaceae) of the Eudicots which are characterized by simultaneous cytokinesis. The occurrence of Ubisch bodies (orbicules) in several species of Apinagia and Marathrum parallels the echinate ornamentation of the pollen grains.     

STM/BP-Like KNOXI Is Uncoupled from ARP in the Regulation of Compound Leaf Development in Medicago truncatula

Class I KNOTTED-like homeobox (KNOXI) genes are critical for the maintenance of the shoot apical meristem. The expression domain of KNOXI is regulated by ASYMMETRIC LEAVES1/ROUGHSHEATH2/PHANTASTICA (ARP) genes, which are associated with leaf morphology. In the inverted repeat-lacking clade (IRLC) of Fabaceae, the orthologs of LEAFY (LFY) function in place of KNOXI to regulate compound leaf development. Here, we characterized loss-of-function mutants of ARP (PHAN) and SHOOTMERISTEMLESS (STM)- and BREVIPEDICELLUS (BP)-like KNOXI in the model IRLC legume species Medicago truncatula. The function of ARP genes is species specific. The repression of STM/BP-like KNOXI genes in leaves is not mediated by PHAN, and no suppression of PHAN by STM/BP-like KNOXI genes was observed either, indicating that STM/BP-like KNOXI genes are uncoupled from PHAN in M. truncatula. Furthermore, comparative analyses of phenotypic output in response to ectopic expression of KNOXI and the M. truncatula LFY ortholog, SINGLE LEAFLET1 (SGL1), reveal that KNOXI and SGL1 regulate parallel pathways in leaf development. We propose that SGL1 probably functions in a stage-specific manner in the regulation of the indeterminate state of developing leaves in M. truncatula.     

Developmental morphology of seedling and shoot and phylogenetic relationship of Diplobryum koyamae (Podostemaceae)

We demonstrated that saltational evolution was recurrent in the body plans of seedlings of the aquatic angiosperm Podostemaceae, in contrast to other angiosperms with seedlings having almost common body plans. Diplobryum koyamae, transferred to the genus Hydrodiscus described in this paper, has long-floating shoots with an anchoring disk-like base and is rootless. Such a body plan is distinct from other members of Asian Podostemoideae comprising reduced or moderate shoots borne on the root. Here, our molecular phylogenetic analysis revealed that H. koyamae is sister to a crustose-rooted group of Hanseniella, Hydrobryum and Thawatchaia within Asian Podostemoideae. The germinating embryo was devoid of plumule and radicle, and comprised a single cotyledon and a short hypocotyl, which produced an adventitious shoot endogenously. The leaves are formed in the absence of the shoot apical meristem, accompanying the separation of lightly stained cells. Comparison with other species of Asian Podostemoideae having the plumule and the adventitious root in the seedling, along with their phylogenetic relationship, suggests that saltational evolution occurred in the seedling body plan of H. koyamae leading to the extraordinary adult body plan, as in the separate clade of Dalzellia, Indodalzellia, and Indotristicha of the subfamily Tristichoideae.     

Infrafamilial Phylogeny of the Aquatic Angiosperm Podostemaceae Inferred from the Nucleotide Sequences of the matK Gene

Abstract: The infrafamilial relationships of Podostemaceae were deduced from nucleotide sequences of the chloroplast matK gene. The matK phylogenetic analyses show that Podostemaceae are composed of two major clades that correspond to the subfamily Tristichoideae sensu stricto and Weddellina and the subfamily Podostemoideae. Weddellina, which has long been recognized as a member of the Tristichoideae, is sister to the Podostemoideae, supporting the classification that recognized a third subfamily Weddellinoideae. Malaccotristicha malayana and Terniopsis sessilis form a basal clade in Tristichoideae sensu stricto. Tristichoideae show a high morphological diversity and, surprisingly, a close relationship exists between Dalzellia zeylanica and Indotristicha ramosissima, which remarkably differ in their body plans. A few genera defined by particular characters, such as Synstylis and Torrenticola, merge into clades of other larger genera. The Podostemoideae taxa studied are composed of two American clades, an Asian-Australian clade and a Madagascan clade, and may suggest that the subfamily perhaps originated in America and migrated to the Old World.     

Developmental anatomy of the reproductive shoot in <Emphasis Type="Italic">Hydrobryum japonicum</Emphasis> (Podostemaceae)

Podostemaceae are unusual aquatic angiosperms adapting to extreme habitats, i.e., rapids and waterfalls, and have unique morphologies. We investigated the developmental anatomy of reproductive shoots scattered on crustose roots of Hydrobryum japonicum by scanning electron microscopy and using semi-thin serial sections. Two developmental patterns were observed: bracts arise either continuously from an area of meristematic cells that has produced leaves, or within differentiated root ground tissue beneath, and internal to, leaf base scars after an interruption. In both patterns, the bract primordia arise endogenously at the base of youngest bracts in the absence of shoot apical meristem, involving vacuolated-cell detachment to each bract separately. The different transition patterns of reproductive shoot development may be caused by different stages of parental vegetative shoots. The floral meristem arises between the two youngest bracts, and is similarly accompanied by cell degeneration. In contrast, the floral organs, including the spathella, arise exogenously from the meristem. Bract development, like vegetative leaf development, is unique to this podostemad, while floral-organ development is conserved.     

Comparative studies in the morphology of Crenias weddelliana and Maferria indica with reference to Sphaerothylax abyssinica (Podostemaceae: Podostemoideae)

The scope of morphological plasticity of vegetative structures among Podostemoideae (Podostemaceae) is documented for Crenias weddelliana, a neotropical species, Maferria indica, a palaeotropical species, and Sphaerothylax abyssinica, from Kenya, and compared with related taxa. The study highlights intrinsic characters of the widely enigmatic plant body of many species of the subfamily Podostemoideae. These include dorsiventrality of shoots occurring irrespective of gravity, incurvate distichy and one‐sided spirodistichy correlated with shoot dorsiventrality, asymmetric leaves, and several types of positioning of the two prophylls and inflorescence structures. The homogeneity of hairs of the ‘Zeylanidium olivaceum type’ established on the subulate leaves of some Indian species is of taxonomic value. The latter also applies to the stipella (not stipule) on the asymmetric compound leaf in New World species. The morphological data represent a framework of features consistent for the subfamily. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society, 138 , 63–84.     

Neglected features of probable taxonomic value in Podostemaceae: The case of Polypleurum

The paper reports on hitherto largely neglected discriminative characters in Polypleurum stylosum, P. elongatum, P. minus, as well as the problematic P. munnarense (Podostemaceae, subfam. Podostemoideae) from India and Sri Lanka. The characters discussed include dimorphism of floriferous and vegetative shoots, leaf dimorphism, presence of peculiar floating trichomes on vegetative leaves, and presence of silica bodies in root and shoot tissue. In addition, anatomical pecularities (well-developed cuticle covering the ovary and ovules, cell inclusions in the form of droplets, and presence of idioblasts with firm inclusions of unknown compounds) and specific morphological features (presence of an apical septum in the gynoecium) are documented. All these features point to different developmental and physiological properties of the species concerned, thus signaling separate evolutionary pathways. They may help to elucidate phylogenetic relationships at various taxonomic levels and may prove useful for the delimitation of the genus.     

Arabidopsis inflorescence architecture requires the activities of KNOX-BELL homeodomain heterodimers

In flowering plants, post-embryonic development is mediated by the activity of shoot and root apical meristems. Shoot architecture results from activity of the shoot apical meristem (SAM), which initiates primordia, including leaves, internodes and axillary meristems, repetitively from its flanks. Axillary meristems can develop into secondary shoots or flowers. In Arabidopsis, two paralogous BEL1-like (BELL) homeobox genes, PENNYWISE (PNY) and POUND-FOOLISH (PNF), expressed in the SAM, encode DNA-binding proteins that are essential for specifying floral primordia and establishing early internode patterning events during inflorescence development. Biochemical studies show that PNY associates with the knotted1-like homeobox (KNOX) proteins, SHOOTMERISTEMLESS (STM) and BREVIPEDICELLUS (BP). PNY-BP heterodimers are essential for establishing early internode patterning events, while PNY-STM heterodimers are critical for SAM function. In this report, we examined the role of PNY, PNF and STM during development. First, we show that PNF interacts with STM and BP indicating that PNY and PNF are redundant functioning proteins. Inflorescence development, but not vegetative development, is sensitive to the dosage levels of PNY, PNF and STM. Characterization of stm-10, a weak allele in the Columbia ecotype, indicates that STM is also involved in floral specification and internode development. Our examination of the genetic requirements for PNY, PNF and STM demonstrates that these KNOX–BELL heterodimers control floral specification, internode patterning and the maintenance of boundaries between initiating floral primordia and the inflorescence meristem.Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .     

Leaf development in the absence of a shoot apical meristem in Zeylanidium subulatum (Podostemaceae)

* BACKGROUND AND AIMS: The Podostemaceae are a family of unusual aquatic angiosperms that live in rapids and waterfalls. To adapt to such extreme habitats, the family shows unusual morphologies. This study investigated the developmental anatomy of the shoot of Zeylanidium subulatum borne on the prostrate root attached to submerged rock surfaces. * METHODS: Shoots of Z. subulatum were observed under the microscope using resin-sections. * KEY RESULTS: The shoot has no shoot apical meristem (SAM) and, without it, forms leaves distichously dorsiventrally facing the immediately older leaf. A new leaf forms on the adaxial side of a pre-existing leaf and also on the abaxial side of a leaf on flowering shoots. In both cases, the young leaf is endogenous below the older leaf and maintains histological continuity with it. Shortly after internal initiation, the leaf primordia become separate from each other due to cleavage between adjacent leaves of opposite ranks. The cleavage is caused by intercellular separation as well as by degeneration of vacuolated cells. Loss of the SAM is probably linked with the speculated shift of the site of leaf formation to the root. * CONCLUSIONS: The 'shoot' of Z. subulatum is characterized by the absence of a SAM, endogenous leaf formation in the absence of a SAM, cleavage between leaf primordia, and adventitious leaf formations. These innovations occur in some Podostemaceae that have become increasingly adapted to extreme aquatic habitats.     

A complex case of simple leaves: indeterminate leaves co-express ARP and KNOX1 genes

The mutually exclusive relationship between ARP and KNOX1 genes in the shoot apical meristem and leaf primordia in simple leaved plants such as Arabidopsis has been well characterized. Overlapping expression domains of these genes in leaf primordia have been described for many compound leaved plants such as Solanum lycopersicum and Cardamine hirsuta and are regarded as a characteristic of compound leaved plants. Here, we present several datasets illustrating the co-expression of ARP and KNOX1 genes in the shoot apical meristem, leaf primordia, and developing leaves in plants with simple leaves and simple primordia. Streptocarpus plants produce unequal cotyledons due to the continued activity of a basal meristem and produce foliar leaves termed “phyllomorphs” from the groove meristem in the acaulescent species Streptocarpus rexii and leaves from a shoot apical meristem in the caulescent Streptocarpus glandulosissimus. We demonstrate that the simple leaves in both species possess a greatly extended basal meristematic activity that persists over most of the leaf’s growth. The area of basal meristem activity coincides with the co-expression domain of ARP and KNOX1 genes. We suggest that the co-expression of ARP and KNOX1 genes is not exclusive to compound leaved plants but is associated with foci of meristematic activity in leaves.     

The ramification of Apinagia riedelii: A key to the understanding of the plant architecture of Podostemaceae,subfamily Podostemoideae

The study of the ramification pattern of Apinagia riedelii results in a new concept of the architecture of this species, with general implications to members of subfamily Podostemoideae with dithecous leaves. The presence of a subtending leaf below the floriferous shoot proves axillary branching also for species with dithecous leaves. Previous opinions of an unusual ramification mode by subfoliar or non-axillary branching or stem bifurcation in combination with dithecous leaves hitherto pleaded for Podostemoideae is refuted. Moreover, the view of the so-called dithecous leaves with one sheath (theca) at the ventral and one at the dorsal side of the leaf, previously regarded as initially connected with branching, has to be changed. The dithecous leaf arises from the branch and not from the mother shoot axis – as previously believed – and represents the addorsed hypsophyll, i.e., the first leaf (prophyll) of the floriferous branch. This finding leads to the conclusion that the lower sheath of the dithecous leaf is the ventral (not dorsal) sheath pointing to the branch and surrounding its flower bud with a ligule or an ochrea and a hood upon the bud. In this way, the branch and its flower bud become seemingly sunk in the leaf base. At the fusion of leaf basis and shoot results this enigmatic common tissue. The wings of the dorsal (upper) sheath of the dithecous leaf point to the mother shoot axis of the branch. Successive floriferous branches along the main stem disclose the shoot axis of A. riedelii as a monopodium (not sympodium) that develops an anthocladial (foliated) inflorescence in the form of a botrys or a compound botrys, respectively. Since it is generally difficult to define cymose or racemose inflorescences if subtending leaves are absent – which occur in most other species of subfamily Podostemoideae with dithecous leaves – the nature of these inflorescences is discussed anew. The findings on A. riedelii have consequences on our comprehension of the shoot architecture of Podostemoideae.