Floral development and anatomy of Salvadoraceae

Background and Aims

This study is an investigation into the floral development and anatomy of two genera of the small family Salvadoraceae, which belongs to the Brassicales in a clade with Batis and Koeberlinia. Salvadoraceae remains little known, despite its wide distribution in arid areas of the globe. Floral morphological data are scarce, and information on floral anatomy is limited to a single study, although morphological and anatomical characters are now used increasingly as a counterpart of molecular data. There remain a number of controversial morphological questions, such as the fusion of the petals, the number of carpels and the nature of the nectaries.

Methods

Floral anatomy and ontogeny were studied in two species of Salvadora and one species of Dobera. Only for S. persica could a full floral developmental sequence be done.

Key Results

The floral development demonstrates that the ovary of Salvadoraceae is basically bicarpellate and pseudomonomerous with a single locule and parietal placenta. The ovary of Dobera resembles Azima tetracantha in the presence of a false apical septum. Evidence for a staminodial nature of the nectaries is not decisive. In Salvadora petals and stamens are lifted by a short hypanthium.

Conclusions

Salvadoraceae share several morphological and developmental synapomorphies with Batis (Bataceae) and possibly Koeberlinia (Koeberliniaceae), supporting their close relationship as indicated by molecular phylogeny.Key words: Batis, Brassicales, Dobera, Emblingia, floral development, floral anatomy, Koeberlinia, phylogeny, Salvadora, Salvadoraceae, SEM     

Embryology of Koeberlinia (Koeberliniaceae): Evidence for core-Brassicalean affinities

Koeberlinia, comprising a single xerophytic species K. spinosa, had previously been placed in various families, mainly Capparaceae. Current molecular evidence now places it in its own family Koeberliniaceae, thought to be related to the Bataceae/Salvadoraceae among the 17 other families of the Brassicales. We investigated 55 embryological characters of the genus, most of which are not understood yet, and thereby assessed its systematic relationships. Koeberlinia has many embryological features in common with the Capparaceae and seven other core-Brassicalean families (i.e., Brassicaceae, Cleomaceae, Emblingiaceae, Gyrostemonaceae, Pentadiplandraceae, Resedaceae, and Tovariaceae), specifically by possessing a campylotropous ovule with a nonmultiplicative (two-cell-layered) outer integument, reniform seeds with a curved embryo, and a fibrous exotegmen in the mature seed coat. However, Koeberlinia is clearly distinguished from them by a tenuinucellate rather than crassinucellate ovule as previously reported, markedly enlarged apical nucellar epidermal cells, and an "exotestal" seed coat. Embryologically, Koeberlinia resembles neither the Bataceae nor the Salvadoraceae, although only limited embryological data are available for these two families. Embryological evidence thus favors its joining the core Brassicales, but additional molecular analyses and embryological studies on the missing data of the Bataceae and Salvadoraceae are needed for final confirmation of its phylogenetic position.     

The role of growth regulators in the differentiation of flowers and inflorescences

Growth regulators participate in the differentiation of floral parts, determining the developmental path of the respective type of inflorescence. The effect depends on the expression of the peculiarities of floral part differentiation, the recognition of the character of endogenous substances in certain stages and the choice of the suitable regulator for application. In the primitive flower ofPapaver petals and stamens are formed from the peripheral meristem with a lower content of auxins and a higher level of gibberellic substances. The pistil arises later from central tissues with a higher level of auxins and inhibitory substances. The stamens are more sensitive to the higher level of auxin substances, and by a suitable application of GA₃ and BAP they can be transformed into petals; in this way double flower forms arise. In the differentiation of floral parts ofCampanula, Rosa andMelandrium similar regularities assert themselves in time successions, but in another spatial arrangement. Sex differentiation of diclinous flowers ofMelandrium is based on differences in heterochromosomes XY and XX. The rise of the zygomorphic flower ofVeronica is accompanied by a different distribution of endogenous substances which affect the development of petals, stamens and the pistil. The differentiation of flowers in the racemose inflorescence occurs in the acropetal succession, and lateral primordia inCampanula develop into actinomorphic regular flowers, whereas inDigitalis they are zygomorphic and only the terminal flower is peloric. In the initial phases the staminate tassel and the pistillate ear in maize are identical. Earlier differentiation of the terminal pistillate tassel is connected with a higher level of gibberellins and the later development of the lateral pistillate ear is accompanied by the increase in auxin-like substances and inhibitions. Similar correlations were found in the development of staminate catkins and the differentiation of pistillate flowers in terminal buds ofJuglans regia. By the application of auxin-like substances it is possible to achieve the transformation of primordia of the staminate tassel into the pistillate ear in maize or to regulate the number of staminate catkins and pistillate flowers on twigs of the walnut tree. In the capitulum of the sunflower differences arise between peripheral pistillate ray flowers and hermaphrodite tubular ones. By applying GA₃ and BAP the number of ray flowers is increased. If the normal course of inflorescence differentiation is affected with a suitable type of regulator, a range of floral abnormalities appears which permit to assess the intervention in different developmental stages and the reaction of the primordium to the applied type of regulator. Abnormalities also suggest some phylogenetic correlations.     

Flower structure and potential bisexuality in Freycinetia reineckei (Pandanaceae), a species of the Samoa Islands

In Freycinetia reineckei the staminate flower (on the staminate spikes) comprises 3 or 4 (sometimes 2) stamens and a pistillode with 2 (sometimes 4) carpellodes, and the pistillate flower (on the pistillate spikes) is formed of a pistil with 2 (sometimes 4) carpels and of 3 or 4 (sometimes 2) staminodes. This perfect floral homology, also observed in all the other species that were studied with both pistillate and staminate material, strongly suggests that the flower of Freycinetia is basically and potentially bisexual, and may explain the occasional sexual lability and bisexuality of that flower (occurrence of both pistillate and staminate inflorescences, and/or of bisexual inflorescences with bisexual flowers and/or unisexual flowers, on the same individuals) in some species, and also the frequent occurrence of bisexual spikes in this species. These may be partitioned into pistillate, staminate, mixed and sterile zones. In the pistillate zones the flowers have the same aspect and structure as the pistillate flowers. In the staminate zones the flowers generally comprise 3 or 4 (sometimes 2) stamens and a ‘semi-pistil’ some have both stamens and staminodes. The semi-pistils are intermediate between pistils and pistillodes in length, aspect and structure, but always have placentas and ovules. In the mixed zones the flowers are generally formed of a pistil and 3 or 4 (sometimes 2) stamens, and are therefore true hermaphrodite flowers; some have both stamens and staminodes. In the sterile zones the flowers comprise a semi-pistil and 3 or 4 (sometimes 2) staminodes. The staminodes are anatomically very similar to the stamens, especially in the staminate, mixed, and sterile zones, in which they exhibit a wide range of variation in length, aspect and structure. The perfect floral homology as generic character on one hand, and the occasional bisexuality both with and without bisexual flowers and other aspects of sex expression (e.g. occurrence of both pistillate and staminate shoots on the same individuals) in some species on the other hand, seem to indicate that Freycinetia is a basically monoecious, sex changing genus.     

The expression and promoter specificity of the birch homologs for PISTILLATA/GLOBOSA and APETALA3/DEFICIENS

B-function genes determine the identity of petals and stamens in the flowers of model plants such as Arabidopsis and Antirrhinum . Here, we show that a putative B-function gene BpMADS2 , a birch homolog for PISTILLATA , is expressed in stamens and carpels of birch inflorescences. We also present a novel birch gene BpMADS8 , a homolog for APETALA3 / DEFICIENS , which is expressed in stamens. Promoter-GUS analysis revealed that BpMADS2 promoter is active in the receptacle of Arabidopsis flower buds while BpMADS8 promoter is highly specific in mature stamens. BpMADS2 promoter:: BARNASE construct prevented floral organ development in Arabidopsis and tobacco. In birch, inflorescences with degenerated stamens and carpels were obtained. BpMADS8::BARNASE resulted in degeneration of stamens in Arabidopsis and birch causing male sterility. In tobacco, only sepals were developed instead of normal flowers. The results show that the BpMADS2::BARNASE construct can be used to specifically disrupt floral organ development in phylogenetically distant plant species. The stamen-specific promoter of BpMADS8 is a promising tool for biotechnological applications in inducing male sterility or targeting gene expression in the late stamen development.     

The development of pistillate and perfect florets in Xeranthemum squarrosum (Asteraceae)

The formation of capitulum inflorescence with two different types of floret is an interesting issue in floral biology and evolution. Here we studied the inflorescence, floral ontogeny and development of the everlasting herb, Xeranthemum squarrosum, using epi‐illumination microscopy. The small vegetative apex enlarged and produced involucral bracts with helical phyllotaxy, which subtended floret primordia in the innermost whorl. Initiation of floret primordia was followed by an acropetal sequence, except for pistillate peripheral florets. The origin of receptacular bracts was unusual, as they derived from the floral primordia rather than the receptacular surface. The order of whorl initiation in both disc and pistillate flowers included corolla, androecium and finally calyx, together with the gynoecium. The inception of sepals and stamens occurred in unidirectional order starting from the abaxial side, whereas petals incepted unidirectionally from the adaxial or abaxial side. Substantial differences were observed in flower structure and the development between pistillate and perfect florets. Pistillate florets presented a zygomorphic floral primordium, tetramerous corolla and androecium and two sepal lobes. In these florets, two sepal lobes and four stamen primordia stopped growing, and the ovary developed neither an ovule nor a typical stigma. The results suggest that peripheral pistillate florets in X. squarrosum, which has a bilabiate corolla, could be considered as an intermediate state between ancestral bilabiate florets and the derived ray florets.     

A new genus of Sapindaceae from Somalia

The new genus Hirania (Sapindaceae), with the single species H. rosea from south-central Somalia, is decribed and illustrated. It is a shrub with simple, entire leaves, alternate or crowded on short-shoots, and without stipules. The inflorescences are branched with monochasially arranged, unisexual flowers. The flowers (only male ones known) are zygomorphic with five sepals with gland-tipped hairs along margins, four pink, subequal, clawed petals without appendages, and eight, glabrous stamens. The dark purple disk is narrow and tubular and consists of an upper broader segment partly enclosing a lower narrower segment. The closest relative is believed to be the Australian genus Diplopeltis.     

Studies on the Development of Flower in Gastrodia ekata Bl.

The flower of Gastrodia elata B1. grows usually solitary in the bract-axial. The parts of flower are appeared in the following order: sepals, petals, stamens and pestil. In addition, the bases of calyx and corolla unite forming the oblique “floral tube”. Inner three petals, the median lip of the petal is usually larger and different in shape. The lip is three lobes, the marginal and its bases meristem keeps its activity forming lobes marginal tassel, at the base forming one fleshy reniform callus on both sides of lip. A column is united by stamen and style. The stamen is on the top at the back of rostellum in the column. Three stamens primodia of them, only one can developed, on the lateral stamens, during the early stage of flower development, it will not development further and stays on rudimentary, it forming two horn-like protuberances of the clinandrium.     

Floral anatomy of Paepalanthoideae (Eriocaulaceae, Poales) and their Nectariferous structures

BACKGROUND AND AIMS: Eriocaulaceae (Poales) is currently divided in two subfamilies: Eriocauloideae, which comprises two genera and Paepalanthoideae, with nine genera. The floral anatomy of Actinocephalus polyanthus, Leiothrix fluitans, Paepalanthus chlorocephalus, P. flaccidus and Rondonanthus roraimae was studied here. The flowers of these species of Paepalanthoideae are unisexual, and form capitulum-type inflorescences. Staminate and pistillate flowers are randomly distributed in the capitulum and develop centripetally. This work aims to establish a floral nomenclature for the Eriocaulaceae to provide more information about the taxonomy and phylogeny of the family. METHODS: Light microscopy, scanning electron microscopy and chemical tests were used to investigate the floral structures. KEY RESULTS: Staminate and pistillate flowers are trimerous (except in P. flaccidus, which presents dimerous flowers), and the perianth of all species is differentiated into sepals and petals. Staminate flowers present an androecium with scale-like staminodes (not in R. roraimae) and fertile stamens, and nectariferous pistillodes. Pistillate flowers present scale-like staminodes (except for R. roraimae, which presents elongated and vascularized staminodes), and a gynoecium with a hollow style, ramified in stigmatic and nectariferous portions. CONCLUSIONS: The scale-like staminodes present in the species of Paepalanthoideae indicate a probable reduction of the outer whorl of stamens present in species of Eriocauloideae. Among the Paepalanthoideae genera, Rondonanthus, which is probably basal, shows vascularized staminodes in their pistillate flowers. The occurrence of nectariferous pistillodes in staminate flowers and that of nectariferous portions of the style in pistillate flowers of Paepalanthoideae are emphasized as nectariferous structures in Eriocaulaceae.     

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.     

Comparative floral structure and systematics in Chrysobalanaceae s.l. (Chrysobalanaceae, Dichapetalaceae, Euphroniaceae, Trigoniaceae; Malpighiales)

Chrysobalanaceae s.l. , one of the few suprafamilial subclades of Malpighiales that is supported by molecular phylogenetic analyses, and containing Chrysobalanaceae, Dichapetalaceae, Euphroniaceae, and Trigoniaceae, was comparatively studied with regard to floral structure. The subclade is well supported by floral structure. Potential synapomorphies for Chrysobalanaceae s.l. are the following shared features: floral cup; flowers obliquely monosymmetric; sepals congenitally united at base; sepals of unequal size (outer two shorter); fertile stamens concentrated on the anterior side of the flower and sometimes united into a strap; staminodes absent in the posteriormost antepetalous position; anthers extremely introrse, with thecae almost in one plane; endothecium continuous over the dorsal side of the connective; dorsal anther pit; gynoecium completely syncarpous up to the stigma; carpel flanks slightly bulged out transversely and thus carpels demarcated from each other by a longitudinal furrow; flowers with dense unicellular, non-lignified hairs, especially on the gynoecium; light-coloured, dense indumentum on young shoots and inflorescences. Potential synapomorphies for Chrysobalanaceae + Euphroniaceae include: spur in floral cup; clawed petals; lignified hairs on petals; nectary without lobes or scales and mostly annular. Potential synapomorphies for Dichapetalaceae + Trigoniaceae include: special mucilage cells in sepals in mesophyll (in addition to epidermis); anthers almost basifixed; gynoecium synascidiate up to lower style; nectary with lobes or scales and semi-annular.  © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society , 2008, 157 , 249–309.     

The systematic relationships of glucosinolate-producing plants and related families: a cladistic investigation based on morphological and molecular characters

A cladistic analysis is performed using 94 morphological and biochemical characters for 42 genera to compare a phylogeny based on morphological data with those obtained using different genes ( rbc L, atp B, 18S RNA, mat K) or their combination with morphological data, and to understand the floral evolution within the expanded Brassicales (Capparales) relative to Sapindales and Malvales. The tree produced with morphological data is congruent with those obtained from macromolecular studies in obtaining a well-supported glucosinolate-producing clade and an expanded Sapindales. The combined analysis of the morphological and molecular characters is generally well resolved with support for many of the relationships. The inclusion of the fossil taxon Dressiantha demonstrates the value of inserting fossil evidence in phylogenetic analyses. However, the fossil appears to be related to the Anacardiaceae and not to the Brassicales. The core Brassicales are well supported by a number of synapomorphies, although the internal position of Tovariaceae and Pentadiplandraceae is not well resolved. Emblingiaceae appears to be related to Bataceae and Salvadoraceae. Several significant morphological characters are mapped on the combined trees and their evolutionary significance is discussed. Within Brassicales and Sapindales several well supported clades can be recognized which merit ordinal or subordinal status, putting the present orders at a higher level; these include: Tropaeolales, Setchellanthales, Batidales, Brassicales (Brassiciflorae), Burserales, Sapindales and Rutales (Sapindiflorae). The present scheme of affinities within the Brassicales corresponds well with a gradual morphological evolution in the order.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 151 , 453–494.     

Unisexual flower, spikelet, and inflorescence development in monoecious/dioecious Bouteloua dimorpha (Poaceae, Chloridoideae)

Unisexual flowers have evolved repeatedly in the angiosperms. In Poaceae, multiple transitions from bisexual to unisexual flowers are hypothesized. There appear to be at least three distinct developmental mechanisms for unisexual flower formation as found in members of three subfamilies (Ehrhartoideae, Panicoideae, Pharoideae). In this study, unisexual flower development is described for the first time in subfamily Chloridoideae, as exemplified by Bouteloua dimorpha. Scanning electron microscopy (SEM) and anatomy were used to characterize the development of male (staminate) and female (pistillate) flowers, spikelets, and inflorescences. We found the developmental pathway for staminate flowers in B. dimorpha to be distinct from that described in the other three subfamilies, showing gynoecial arrest occurs at a different stage with possible loss of some cellular contents. However, pistillate flowers of B. dimorpha had some similarity to those described in other unisexual-flowered grasses, with filament and anther differentiation in abortive stamens. Comparing our findings with previous reports, unisexual flowers seem to have evolved independently in the four examined grass subfamilies. This analysis suggests the action of different genetic mechanisms, which are consistent with previous observations that floral unisexuality is a homoplasious condition in angiosperms.     

Patterns of flower morphology and structural changes during interconversion between chasmogamous and cleistogamous flowers in Viola philippica

Aims Viola philippica is a species with a typical chasmogamous-cleistogamous (CH-CL) mixed breeding system. It provides a flower model system to investigate floral organs development under different photoperiods. Morphological changes of intermediate cleistogamous (inCL) flowers have been observed, the trends in variation of changes from CH flowers to CL flowers or from CL flowers to CH flowers have been analyzed, the localized effects of poorly developed stamens and petals in CL and inCL flowers have been identified. This research provided morphology and structural changes with implication for the evolutionary significance of the dimorphic flower formation for further study in dimorphic flower development.Methods We used methods of anatomy and structural analysis to observe the morphological structures of flowers under different photoperiods.Important findings Photoperiod played an important role in the development of CH and CL flowers in V. philippica. Under short-day light and intermediate-day light, both CH and inCL flowers developed simultaneously. Most of the floral buds were CH flowers under a photoperiod of short-day light, but most of the floral buds were inCL flowers under mid-day light. Complete CL flowers formed under long-day lights. However, there were a series of transitional types in the number and morphology of stamens and petals among inCL flowers, including five stamens with three petals related to CH flowers and two stamens with one petal related to CL flowers. The former type was dominant under short-day light conditions, and the latter type was dominant under mid-day light. Further more, there were localized effects in stamen and petal development for CL and inCL flowers. The development of ventral lower petal (corresponding to the lower petal with spur of CH flower) and the adjacent two stamens in inCL flowers were best, and the back petal was similar to that of CL flowers, an organ primordium structure. The adjacent stamens with the back petals tended to be poorly developed. In extreme cases, these stamens in inCL flowers had no pollen sac, only a membranous appendage or even a primordium structure. When the plants with CL or CH flowers were placed under short-day light or long-day light, the newly induced flowers all showed a series of inCL flower types, finally the CL flowers transformed into CH flowers, and the CH flowers transformed into CL flowers. This result indicates the gradual effects of different photoperiods on dimorphic flowers development of V. philippica. A long photoperiod could inhibit the development of partial stamens and petals, and a short photoperiod could prevent the suppression of long-day light and promote the development of stamens and petals.     

Pistillate and staminate flower development in dioecious Pistacia vera (Anacardiaceae)

The development of staminate and pistillate flowers in the dioecious tree species Pistacia vera L. (Anacardiaceae) was studied by scanning electron microscopy with the objective of determining organogenetic patterns and phenology of floral differentiation. Flower primordia are initiated similarly in trees of both sexes. Stamen and carpel primordia are initiated in both male and female flowers, and the phenology of organ initiation is essentially identical for flowers of both sexes. Vestigial stamen primordia arise at the flanks of pistillate flower apices at the same time functional stamens are initiated in the staminate flowers. Similarly, a vestigial carpel is initiated in staminate flowers at the same time the primary, functional carpel is initiated in pistillate flower primordia. Differences between the two sexes become apparent early in development as, in both cases, development of organs of the opposite sex becomes arrested at the primordial stage. Male flowers produce between four and six mature functional stamens and female flowers produce a gynoecium with one functional and two sterile carpels.     

On the morphological position of Paepalanthus subgenus Psilandra (Eriocaulaceae)

Paepalanthus subgen. Psilandra comprises two species known only from the type collections. The aim of this study is to evaluate their unique floral morphology within a given phylogenetic context and its implications for taxonomy. Flowers of both species were analyzed, described, and photographed using a digital microscope. Taxonomically relevant characters were mapped on previous molecular phylogenies. The flowers of both species fit the Paepalanthoideae pattern and are quite similar to each other. The petals of the pistillate flowers, although free, show evidence of previous fusion in the middle. The free condition is interpreted to be secondary, probably because of the handling of the specimens or to the current developmental stage. Character mapping indicates that both species share floral features with Syngonanthus but not with Paepalanthus. Therefore, based on the median fusion of the pistillate flower petals, absence of floral bracts, petals of the pistillate flower being shorter than the sepals, simple stigmas in the commissural position, the pedicellate pistillate flower, and the glabrous petals and sepals, we transfer P. subgen. Psilandra to synonymy of Syngonanthus sect. Syngonanthus and combine both species in this genus.     

Reproductive structures of an extinct platanoid from the early Cretaceous (latest Albian) of eastern North America

Two new species of platanoid reproductive structure are described from the Bull Mountain locality in the Patapsco Formation (Potomac Group) of northeastern Maryland, USA. Pistillate inflorescences and infructescences (Platanocarpus elkneckensis sp. nov.) consist of flowers and fruits in sessile globose heads that are borne on an elongate axis. Individual pistillate flowers consist of five free carpels surrounded by prominent tepals. Staminate inflorescences, flowers and isolated stamens are assigned to Hamatia elkneckensis gen. et sp. nov. Staminate flowers are borne in a globose head with a small number of stamens (five?) per flower. Stamens consist of very short filaments, long anthers with strongly valvate dehiscence and an apically expanded connective. The connective expansion is frequently very well-developed, hook-like and extends down the ventral surface of the stamen. Anthers contain small, tricolporate, reticulate pollen. Association evidence, similarity of inflorescence structure and the occurrence of Hamatia-type pollen on flowers, carpels and fruits of Platanocarpus elkneckensis suggests that the staminate and pistillate material was produced by a single species of plant. The “Hamatia-plant” provides further evidence of pentamerous floral structure in mid-Cretaceous platanoids and documents the occurrence of unequivocal tricolporate pollen in the platanoid complex.     

Flower morphology and biology of Myrsine laetevirens, structural and evolutionary implications of anemophily in Myrsinaceae

This study deals with the phenology, pollination biology and floral morphology of Myrsine laetevirens , a neotropical dioecious tree. In Punta Lara (Argentina), its flowering period occurs during January-February. Both pistillate and staminate flowers are small, with a yellowish-green perianth and produce neither nectar nor odour. Staminate flowers have five stamens and a vestigial gynoecium while pistillate flowers, have non-functional anthers and a conspicuous stigma. The floral characteristics of staminate and pistillate plants are related to the syndrome of anemophily. Fruit set in inflorescences covered with mesh bags and observations prove that animals are not involved in the pollination process. A provisional cladistic analysis of Myrsinaceae shows that dioecy evolved as one of the most recent apomorphies of Myrsine and is part of the anemophilous syndrome.     

Floral development of the model legume <Emphasis Type="Italic">Medicago truncatula</Emphasis>: ontogeny studies as a tool to better characterize homeotic mutations

This work provides new evidence of the complex genetic regulation necessary to accomplish flower development in legumes. Using scanning electron microscopy (SEM) analysis, we have characterized the early developmental events of the wild type Medicago truncatula flower and selected morphological characters as markers to break it down into eight different developmental stages. The order of floral organ initiation in M. truncatula and pea (Pisum sativum L.), in contrast to Arabidopsis and Antirrhinum, is unidirectional in all whorls starting from the abaxial position of the flower with a high degree of overlap. Another main difference is the existence of four common primordia from which petals and stamens differentiate. The formation of common primordia, as opposed to discrete petal and stamen primordia, has been described in many legume and non-legume plants. The main differences between pea and M. truncatula floral ontogeny are in carpel and fruit development. We also used these morphological markers as tools to characterize early alterations in the flower development of a male-sterile M. truncatula floral homeotic mutant named mtapetala. This mutant displays a phenotype resembling those of weak class B mutants with homeotic conversions of floral organ whorls 2 and 3 into sepaloid and carpelloid structures, respectively. Ontogeny studies of the mtapetala mutant flowers showed similarities with the effects of previously described loss-of-B-function mutations. Differences between ontogeny of wild type and mtapetala flowers could not be detected during the first stages (1-5) of flower development. In late stage 5, abnormal-shaped petals with acute lobes and trichomes as well as abnormal-shaped stamens were visible in whorls 2 and 3. At stage 6, the morphology of petals began to change, developing enlarged sepaloid structures bearing trichomes and first the antesepalous stamens and then the antepetalous stamens began to differentiate carpelloid anthers from filaments. Third whorl organs presented different degrees of carpelloidy. The present study should provide tools for the characterization and comparative analyses of new Medicago floral homeotic mutants and could be useful in elucidating how floral organ identity functions work in legumes.