Molecular separation of genera in Cassiinae (Leguminosae), and analysis of variation in the nodulating species of Chamaecrista

Randomly Amplified Polymorphic DNA (RAPD) methods have been adapted for use as a phenetic tool on the legume tribe Cassiinae. RAPD-generated polymorphism within local populations was lower than between populations from different geographic regions, between species and genera. Examination of three Cassia species, 12 Chamaecrista species and 13 Senna species using eight primers showed the potential for separation of the nodulated/nitrogen fixing genus Chamaecrista from the previously congeneric groups Cassia and Semis. Similarly, RAPD analysis of two groups of nine Ch. rotundifolia and nine Ch. mimosoides samples using 11 primers has given separation according to both species and to geographical location. Analysis of a small sample of five Chamaecrista species from Brazil with eight primers gave separation consistent with known variations in nodule structure.     

Floral ontogeny in legume genera Petalostylis, Labichea, and Dialium (Caesalpinioideae: Cassieae), a series in floral reduction

Floral ontogeny of taxa of two subtribes (Labicheinae, Dialiinae) of caesalpinioid tribe Cassieae, characterized by reduced number of floral organs, was compared. All three taxa studied are distichous; Petalostylis labicheoides flowers are solitary in leaf axils, Labichea lanceolata has few-flowered racemes, and Dialium guineense has numerous-flowered cymes. The first sepal primordium in each is initiated abaxially and nonmedianly. Order of organogenesis in Petalostylis is: five sepals bidirectionally, five petals and carpel simultaneously, then five stamens bidirectionally, starting abaxially. The order in Labichea is: five sepals helically (one lagging in time), five petals unidirectionally starting abaxially, the carpel and petals concurrently, then two stamens successively, starting laterally. Order in Dialium is: five sepals bidirectionally, the single petal adaxially, and lastly the carpel and two stamens concurrently. Specializations include (1) reduction of the five sepals to four by fusion in Petalostylis and Labichea; (2) reduction of petal number to one in Dialium; (3) reduction of stamen number to two in Labichea and Dialium, and reduction of functional stamens to three in Petalostylis; and (4) an elaborate, late-developing style in Petalostylis. Floral asymmetry, another specialization, characterizes Labichea, expressed by dissimilar stamens, while the other genera have zygomorphic flowers. Floral ontogenies are compared with other taxa of Cassieae.     

Comparative floral development and evolution in tribe Caesalpinieae (Leguminosae: Caesalpinioideae). Haematoxylum

Floral organogenesis and development of the tropical legume trees Haematoxylum campechianum (logwood) and H. brasiletto (brazilwood) were studied using scanning electron microscopy. The aims were to compare ontogenies, and to elucidate the relationships of Haematoxylum with other genera of Caesalpinieae, the basal tribe of Caesalpinioideae. Flowers of Haematoxylum are in racemes or fascicles, lack bracteoles, and are pentamerous, hermaphroditic, and either actinomorphic or zygomorphic. Whorls arise in acropetal order except for the carpel, which arises concurrently with the outer stamens. Sepal order is bidirectional (a rare condition) within the whorl in both. Petals and outer stamens are initiated bidirectionally in H. campechianum, and unidirectionally in H. brasiletto. Inner stamens are initiated unidirectionally in both. In H. campechianum, time of petal initiation overlaps with that of outer stamens, and initiation of the two stamen whorls overlap. In both, the gynoecium becomes stipitate, and a hypanthium forms late in development. Both show many plesiomorphic states at anthesis; H. brasiletto alone shows several specialized states (expressed late in development), including a fused, gibbous calyx cup, a zygomorphic corolla, lightly aggregated filaments held together by hairs, and fenestrations in the stamen column. Ontogenetic divergence late in ontogeny characterizes differences at anthesis between related species.     

Seed ontogeny of Chamaecrista and its systematic implications in Cassiinae (Leguminosae, Caesalpinioideae)

The genus Chamaecrista originates from the dismembering of Cassia L., which was elevated to the level of subtribe Cassiinae, giving rise to Cassia sensu stricto, Chamaecrista, and Senna. Phylogenetic analysis has suggested that Cassiinae is not monophyletic. The relationships between its genera are not clear, because they sometimes appear close to Chamaecrista and Senna and at other times close to Senna and Cassia. We describe here development of the seeds of three species of Chamaecrista and compare our data with information published in the literature for Cassiinae and non-Cassiinae legumes. The ovules appear well conserved in Cassiinae, differing only in the length of the funiculus. In terms of seed development, the shape of the endosperm haustorium is similar in Senna and Chamaecrista. Chamaecrista shares more characteristics of its mature seeds with Senna than with Cassia. Chamaecrista and Senna have variable seed shapes, the presence of pleurograms, the absence of root cap primordia, a short epicotyl, and the cotyledonary mesophyll tends to be dorsiventral. These seed characteristics place Senna close to Chamaecrista and distance it from Cassia. We also observed characteristics unique to Chamaecrista, for example the occurrence of multiple pleurograms, a partially multiplicative seed coat, and the presence of embryonic colleters.     

A phenetic study of Cassia sensu lato (Leguminosae-Caesalpinioideae: Cassieae: Cassiinae) in Thailand

Cassia L. sensu lato, a large heterogeneous genus of flowering plants, occurs naturally in the tropics around the world. Recent works based on floral morphology have supported a division of this genus into three genera, namely Cassia L. s. str., Chamaecrista Moench and Senna Mill. In order to investigate this new classification, 508 specimens of 18 taxa of the genus Cassia s.l. grown in Thailand were analyzed using cluster analysis and canonical discriminant analysis. The total 32 vegetative and reproductive morphological characters were employed in these analyses. In cluster analysis, Cassia s.l. can be separated into four groups, respectively viz. Chamaecrista, Senna alata, Senna and Cassia s /str. The four-cluster grouping is discussed. From a canonical discriminant analysis using the four-cluster grouping as a priori groups, it can be concluded that Cassia s. str., Senna, and Chamaecrista are indeed distinct taxa. The three most important characters that separate the three genera are filament length, fruit length, and ovary stalk length. These quantitative characters, together with some qualitative characters, were useful in constructing an identification key to these genera. Among the three genera, it was also found that Senna is rather a heterogeneous taxon. The difference between the studied species was discussed.     

Patterns and development of floral asymmetry in Senna (Leguminosae, Cassiinae)

The buzz-pollinated genus Senna (Leguminosae) is outstanding for including species with monosymmetric flowers and species with diverse asymmetric, enantiomorphic (enantiostylous) flowers. To recognize patterns of homology, we dissected the floral symmetry character complex and explored corolla morphology in 60 Senna species and studied floral development of four enantiomorphic species. The asymmetry morph of a flower is correlated with the direction of spiral calyx aestivation. We recognized five patterns of floral asymmetry, resulting from different combinations of six structural elements: deflection of the carpel, deflection of the median abaxial stamen, deflection or modification in size of one lateral abaxial stamen, and modification in shape and size of one or both lower petals. Prominent corolla asymmetry begins in the earl-stage bud (unequal development of lower petals). Androecium asymmetry begins either in the midstage bud (unequal development of thecae in median abaxial stamen; twisting of androecium) or at anthesis (stamen deflection). Gynoecium asymmetry begins in early bud (primordium off the median plane, ventral slit laterally oriented) or midstage to late bud (carpel deflection). In enantiostylous flowers, pronouncedly concave and robust petals of both monosymmetric and asymmetric corollas likely function to ricochet and direct pollen flow during buzz pollination. Occurrence of particular combinations of structural elements of floral symmetry in the subclades is shown.     

Floral development in Tribe Detarieae (Leguminosae: Caesalpinioideae): Amherstia, Brownea, and Tamarindus

Floral development was compared among three taxa in caesalpinioid tribe Detarieae sensu lato: Amherstia nobilis and Tamarindus indica have racemose, helically arranged inflorescences, while Brownea latifolia has cauliflorous capitate flower clusters that arise as racemes. All have acropetal flower order; initiation and development are sequential in all except Brownea, which is synchronous. All have paired persistent showy bracteoles. Floral symmetry is dorsiventral (zygomorphic) in all except Brownea, with radial symmetry at anthesis. Sepals initiate helically on a circular floral apex, starting with a median abaxial sepal, in all. Petals are initiated helically in Brownea, and unidirectionally in Amherstia and Tamarindus. Stamens are initiated unidirectionally in each stamen whorl in all except Amherstia, in which the outer whorl is bidirectional. The carpel initiates concurrently with the petals in Brownea, and with the outer stamens in the other taxa. The two upper (adaxial) sepal primordia become fused during development in all, so that the calyx appears tetramerous. Some reduced petals occur in Amherstia and Tamarindus, and some reduced stamens occur in all. All produce a hypanthium by zonal growth, and all except Tamarindus have the gynoecium attached adaxially to the hypanthial rim.     

Cytogenetics and chromosome banding patterns in Caesalpinioideae and Papilionioideae species of Pará, Amazonas, Brazil

Despite the abundance of woody legumes in Brazilian Amazonian rain forests, there are few chromosome counts on the native species of this important region. The present work presents such data for 13 species of Caesalpinioideae (the genera Bauhinia , Caesalpinia , Cassia , Chamaecrista and Senna ) and Papilionioideae ( Bowdichia , Centrosema and Dioclea ) collected from 17 natural populations. Our report represents the first chromosome counts for the genera Bowdichia and Dioclea and for four of the studied species. Observations are made on chromosome morphology, size, condensing behaviour and interphase nucleus structure. Banding with fluorochromes carried out for the first time in Caesalpinioideae revealed discrete CMA⁺/DAPI^– terminal (GC-rich) bands on 2–4 chromosome pairs of most species analysed, with a few species presenting discrete CMA^–/DAPI⁺ (AT-rich) bands. Significant differences in chromosome size, morphology and condensing behaviour were observed among members of the controversial tribe Cassieae ( Cassia , Chamaecrista and Senna ), revealing the tribe to be a heterogeneous group from the karyological point of view.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 144 , 181–191.     

Floral ontogeny in Dialiinae (Caesalpinioideae: Cassieae), a study in organ loss and instability

The Caesalpinioideae are widely variable in their floral ontogeny, and among caesalpinioids, members of the polyphyletic tribe Cassieae are particularly diverse. Within the Cassieae, the monophyletic Dialiinae clade is also marked by a high degree of organ loss, particularly in the largest genus, Dialium. The purpose of this work is to explore the ontogeny of several previously undocumented species of the diverse Dialiinae clade, with the goal of building a more complete picture of floral development and evolution in this group and especially within Dialium. We have documented the floral ontogeny of six species of the Dialiinae; four from Dialium, as well as Poeppigia procera and Mendoravia dumaziana. Mode and timing of organ initiation were mostly consistent across the Dialium species studied. With the exception of Dialium dinklagei, which undergoes helical calyx initiation, all flowers initiated sepals bidirectionally. In the instances of both gains and losses of floral organs in Dialium, one trend is apparent — an absence of abaxial organs. Gains in both sepals and stamens occur in the adaxial median position, while stamens and petals which are lost are always the ventral-most organs. Organ initiation in Poeppigia and Mendoravia is unlike that seen in Dialium. Poeppigia shows a ventral to dorsal unidirectional sepal initiation, while both Poeppigia and Mendoravia display near-synchronous initiation of the corolla and staminal whorls. The taxa examined here exemplify the apparent lack of developmental canalisation seen in caesalpinioid legumes. This ontogenetic plasticity is reflective of the morphological diversity shown by flowers across the subfamily, representing what has been described as an “experimental” phase in legume floral evolution.     

Phylogenetic relationships within Senna (Leguminosae, Cassiinae) based on three chloroplast DNA regions: patterns in the evolution of floral symmetry and extrafloral nectaries

Senna (Leguminosae) is a large, widespread genus that includes species with enantiostylous, asymmetric flowers and species with extrafloral nectaries. Clarification of phylogenetic relationships within Senna based on parsimony analyses of three chloroplast regions (rpS16, rpL16, and matK) provides new insights on the evolution of floral symmetry and extrafloral nectaries. Our results support the monophyly of only one (Psilorhegma) of the six currently recognized sections, while Chamaefistula, Peiranisia, and Senna are paraphyletic, and monotypic Astroites and Paradictyon are nested within two of the seven major clades identified by our molecular phylogeny. Two clades (I, VII) include only species with monosymmetric flowers, while the remaining clades (II-VI) contain species with asymmetric, enantiostylous flowers, in which either the gynoecium alone or, in addition, corolla and androecium variously contribute to the asymmetry. Our results further suggest that flowers were ancestrally monosymmetric with seven fertile stamens and three adaxial staminodes, switched to asymmetry later, and reverted to monosymmetry in clade VII. Fertility of all 10 stamens is a derived state, characterizing the Psilorhegma subclade. Extrafloral nectaries evolved once and constitute a synapomorphy for clades IV-VII ("EFN clade"). These nectaries may represent a key innovation in plant defense strategies that enabled Senna to undergo large-scale diversification.     

A striking new unifoliolate and polycarpellate species of Senna (Fabaceae: Caesalpinioideae) from Andhra Pradesh,India

Senna andhrica P.V. Ramana, J. Swamy and M. Ahmedullah, a new species from Andhra Pradesh, India, is described. The species is remarkable in having unifoliolate, oblong or ovate leaves, 2–3 carpellate gynoecium and reniform seeds. It is the second described species with unifoliolate leaves in the Cassiinae. In most other morphological characters, it is closely allied to Senna occidentalis (L.) Link.     

Enantiostyly in Chamaecrista ramosa (Fabaceae‐Caesalpinioideae): floral morphology,pollen transfer dynamics and breeding system

Enantiostyly is a form of reciprocal herkogamy, in which floral morphs present reciprocal differences in the position of sexual elements, and occurs in monomorphic and dimorphic forms. This polymorphism maximises cross‐pollination and reduces self‐pollination, being very common within the subtribe Cassiinae (Fabaceae). Nevertheless, few studies have investigated the functionality of enantiostyly, particularly in this plant group. The present study aimed to investigate enantiostyly and its functionality in Chamaecrista ramosa, a monomorphic enantiostylous shrub, in an area of coastal vegetation in northeast Brazil. Pollen deposition and capture on the body of floral visitors, the relationship of these data with floral biology and breeding system, and morph ratio were evaluated. Pollen deposition and capture occurred in specific sites of the floral visitor body, showing the functionality of enantiostyly. The floral architecture, associated with the floral visitor behaviour, resulted in indirect pollen deposition on the floral visitor body. This occurred through a loop made by the pollen upon the inner petal surface, similar that generally reported for other Cassiinae. Chamaecrista ramosa is self‐compatible, although no fruit set was observed through spontaneous self‐pollination. The occurrence and number of floral morphs was similar within clumps. Enantiostyly seems to be advantageous for this species, as it results in efficient pollen capture and deposition, reduces the chances of autogamy and maximises intermorph pollen flow.     

Taxonomic and functional implications of stigma morphology in species ofCassia,Chamaecrista, andSenna (Leguminosae:Caesalpinioideae)

Two stigma forms occur inChamaecrista andSenna, but only one inCassia. In the common chambered form, a stigma pore is positioned on the reflexed style tip and is the entrance to a tapering chamber. The pore rim is fringed by hairs which vary in number, size, distribution and shape. In the alternative form the stigma is situated at the apex of the curved style and is crateriform. The crater rim is fringed by hairs of variable number and shape. The stigmatic hairs are predominantly unicellular and cutinized. Stigma and hair differences aid in the taxonomy of the genera. Their functions in pollination biology are discussed.     

Development of megagametophyte, embryo, and seed in Senna corymbosa (Lam.) H.S. Irwin & Barneby (Leguminosae – Caesalpinioideae)

The main aspects of seed ontogeny in Senna corymbosa were studied by standard anatomical microtechniques for light microscope observations. The results revealed an ana-campylotropous, bitegmic, and crassinucelate mature ovule. A single archesporocyte developed by an archesporial cell enlargement from the subhypodermal multicellular archesporium. Meiosis originated linear or T-shaped megasporic tetrads. The functional megaspore was the chalazal one. Megagametophytic development conformed to the Polygonum type. Fertilization was porogamic. Endosperm development was free nuclear and conformed to a chalazal haustorium. Cellular endosperm was initiated from the micropylar end during the globular embryo stage. Embryogeny derived from a linear proembryonal tetrad. The mature embryo showed an oblique axis. The testa derived from the outer ovular integument. Nucellar and endosperm remnants, and the micropylar region of the inner ovular integument, persisted at embryo maturity. The absence of a pleurogram would be adaptative to wetland habitats. The taxonomic use of the mature embryo axis in the Cassieae and the phylogenetic employment of megasporic arrangements in Leguminosae needs some reinterpretation.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 169–179.     

Comparative floral development in the tribe Mentheae (Nepetoideae: Lamiaceae) and its bearing on the evolution of floral patterns in asterids

A comparative developmental study of flowers was carried out using epi-illumination light microscopy on four genera of Lamiaceae (Nepeta, Rosmarinus, Salvia, andZiziphora), representing all three subtribes of Mentheae. All species examined share unidirectional (adaxial to abaxial) sepal initiation, except Rosmarinus, which has the reverse unidirectional sequence, starting abaxially. Initiated but suppressed bracteoles were detected only in Rosmarinus. In Rosmarinus, Salvia, and Ziziphora, initiation of petals and stamens proceeds unidirectionally from the abaxial side. Floral initiation of Nepeta has bidirectional inception of petals and unidirectional stamen initiation from the adaxial side. Temporal overlap in organ initiation between petal and stamen whorls occurs in all taxa, though this feature is more prominent in Rosmarinus. Significant structural and developmental features that distinguish the four genera include: (1) polysymmetric calyx tube, highly tomentose corolla and deeply four-partitioned ovary in Nepeta; (2) monosymmetric two-lipped calyx and shallowly four-partitioned ovary in Ziziphora; and (3) suppression of adaxial stamens in Salvia and Rosmarinus. Adaxial stamens are absent from Rosmarinus, but reduced stamens remain as staminodia in Salvia. In a phylogenetic context, the late monosymmetry of Nepeta and very early monosymmetry of Rosmarinus could both be regarded as derived conditions compared with the early monosymmetry ofSalvia and Ziziphora.     

The questionable relationship of Montinia (Montiniaceae): evidence from a floral ontogenetic and anatomical study

The systematic position of Montiniaceae remains uncertain: a relationship with Cornales has been suggested on phytochemical and embryological evidence, while molecular data point to a relationship with Solanales. We investigated the floral development and anatomy of the South African Montinia caryophyllacea to add a new set of characters for clarifying the systematic position of the family Montiniaceae. Pistillate inflorescences show a higher degree of reduction than staminate, with flowers set terminally on short lateral branches. Flowers have an irregular initiation sequence, with frequent abortions of organs. In Montinia, petals grow rapidly, and no zonal growth takes place. The gynoecium develops as a pit surrounded by a girdle. Placentation is basically parietal and becomes axillary by the postgenital fusion of placental lobes; unitegmic ovules are arranged in two parallel rows with adjacent ovules partly overlapping each other. Unisexuality is respectively attained at the stage of anther development and carpel initiation. The floral anatomy of pistillate and staminate flowers is illustrated and discussed. Observations on Montinia are compared with data of taxa from Saxifragaceae sensu stricto, Cornales, and Solanales. The absence of sympetaly in Montinia is discussed. Morphological and anatomical evidence points to a high similarity with Escalloniaceae. Although a position in the asterids is most probable, there is little support for the relationship with Solanales indicated by molecular data.     

The floral scales in Hellmuthia (Cyperaceae, Cyperoideae) and Paramapania (Cyperaceae, Mapanioideae): an ontogenetic study

BACKGROUND AND AIMS: In 1976 the monotypic genus Hellmuthia was placed in the Hypolytreae s.l., but was subsequently ascribed to the Mapanioideae, tribe Chrysitricheae, mainly because of the presence in Hellmuthia of two lateral, mapanioid-like floral scales with ciliated keels, the anatomy of the nutlet, the embryo and the inflorescence. Recently, based on cladistic analyses and supported by pollen ontogenetic evidence, Hellmuthia was transferred to a Cyperaceae, tribe Cypereae, clade mainly consisting of Ficinia and Isolepis. In this study, the floral ontogeny in Hellmuthia was investigated and compared with the floral ontogeny in Paramapania, with special attention for the floral scales. METHODS: Freshly collected inflorescences of Hellmuthia membranacea and Paramapania parvibractea were investigated using scanning electron and light microscopy. KEY RESULTS: In the conical 'spikelet' in Hellmuthia, proximal bracts occur, each axillating an axis with empty glume-like structures, or a reduced spikelet. Hence, it is a reduced partial inflorescence. In Hellmuthia, the stamen primordia originate before the primordia of the perianth-gynoecium appear. Moreover, a third adaxially positioned 'floral scale' was observed for the first time. The position and relative time of appearance of the floral scales in Hellmuthia are typical for perianth parts in Cyperoideae. The basal position of Hellmuthia within a clade of species with usually perianthless flowers, allows the presence of rudiments of a perianth in Hellmuthia to be interpreted as a primitive state. Development of the lateral 'scales' in Paramapania follows a different pattern. Therefore, it was decided that the lateral 'scales' in Paramapania are different from the lateral perianth parts in Hellmuthia. The pollen grains in Hellmuthia are cyperoid, with one polar and five lateral apertures, of which the membrane is covered with sexinous bodies. The pollen surface is granulate and perforate with microspines. CONCLUSIONS: The floral ontogeny in Hellmuthia occurs according to the general cyperoid pattern. The lateral scales in Hellmuthia are perianth parts, and they are not homologous to the lateral 'scales' in Paramapania.     

Seed of Cercis siliquastrum L (Leguminosae,Caesalpinioideae): Reserves and their Utilization after Germination

An ultrastructural study on the reserves stored in cotyledons and endosperm and on their utilization after germination has been carried out in Cercis siliquastrum seeds. The observations have shown that all the cotyledonal reserves of proteins and lipids are used during seedling growth. However, inside the endosperm cells only the protein reserves are depleted, and large quantities of lipids remain undigested. In both cell walls and intercellular spaces of the endosperm cells huge masses of polysaccharide reserves are stored. However, only the polysaccharides inserted in the cell walls are dismantled, while those located in the intercellular spaces are not attacked by the lytic enzymes. Some hypotheses regarding the possible roles of those endospermic non-nutritional materials in the strategies of seed germination and seedling growth are suggested.