Male Flower Development Research Articles

Comparative transcriptomic analysis of male and female flowers of monoecious Quercus suber.

Monoecious species provide a comprehensive system to study the developmental programs underlying the establishment of female and male organs in unisexual flowers. However, molecular resources for most monoecious non-model species are limited, hampering our ability to study the molecular mechanisms involved in flower development of these species. The objective of this study was to identify differentially expressed genes during the development of male and female flowers of the monoecious species Quercus suber, an economically important Mediterranean tree. Total RNA was extracted from different developmental stages of Q. suber flowers. Non-normalized cDNA libraries of male and female flowers were generated using 454 pyrosequencing technology producing a total of 962,172 high-quality reads with an average length of 264 nucleotides. The assembly of the reads resulted in 14,488 contigs for female libraries and 10,438 contigs for male libraries. Comparative analysis of the transcriptomes revealed genes differentially expressed in early and late stages of development of female and male flowers, some of which have been shown to be involved in pollen development, in ovule formation and in flower development of other species with a monoecious, dioecious, or hermaphroditic sexual system. Moreover, we found differentially expressed genes that have not yet been characterized and others that have not been previously shown to be implicated in flower development. This transcriptomic analysis constitutes a major step toward the characterization of the molecular mechanisms involved in flower development in a monoecious tree with a potential contribution toward the knowledge of conserved developmental mechanisms in other species.

Ectopic expression of CsCTR1, a cucumber CTR-like gene, attenuates constitutive ethylene signaling in an Arabidopsis ctr1-1 mutant and expression pattern analysis of CsCTR1 in cucumber (Cucumis sativus).

The gaseous plant hormone ethylene regulates many aspects of plant growth, development and responses to the environment. Constitutive triple response 1 (CTR1) is a central regulator involved in the ethylene signal transduction pathway. To obtain a better understanding of this particular pathway in cucumber, the cDNA-encoding CTR1 (designated CsCTR1) was isolated from cucumber. A sequence alignment and phylogenetic analyses revealed that CsCTR1 has a high degree of homology with other plant CTR1 proteins. The ectopic expression of CsCTR1 in the Arabidopsis ctr1-1 mutant attenuates constitutive ethylene signaling of this mutant, suggesting that CsCTR1 indeed performs its function as negative regulator of the ethylene signaling pathway. CsCTR1 is constitutively expressed in all of the examined cucumber organs, including roots, stems, leaves, shoot apices, mature male and female flowers, as well as young fruits. CsCTR1 expression gradually declined during male flower development and increased during female flower development. Additionally, our results indicate that CsCTR1 can be induced in the roots, leaves and shoot apices by external ethylene. In conclusion, this study provides a basis for further studies on the role of CTR1 in the biological processes of cucumber and on the molecular mechanism of the cucumber ethylene signaling pathway.

Identification of floral genes for sex determination in Calamus palustris Griff. by using suppression subtractive hybridization.

Calamus palustris Griff. is an economically important dioecious rattan species in Southeast Asia. However, dioecy and onset of flowering at 3-4 years old render uncertainties in desired female:male seedling ratios to establish a productive seed orchard for this rattan species. We constructed a subtractive library for male floral tissue to understand the genetic mechanism for gender determination in C. palustris. The subtractive library produced 1536 clones with 1419 clones of high quality. Reverse Northern screening showed 313 clones with differential expression, and sequence analyses clustered them into 205 unigenes, including 32 contigs and 173 singletons. The subtractive library was further validated with reverse transcription-quantitative polymerase chain reaction analysis. Homology identification classified the unigenes into 12 putative functional proteins with 83% unigenes showing significant match to proteins in databases. Functional annotations of these unigenes revealed genes involved in male flower development, including MADS-box genes, pollen-related genes, phytohormones for flower development, and male flower organ development. Our results showed that the male floral genes may play a vital role in sex determination in C. palustris. The identified genes can be exploited to understand the molecular basis of sex determination in C. palustris.

Floral structure and organogenesis of the wax palm Ceroxylon ceriferum (Arecaceae; Ceroxyloideae)

Most palm systematists were surprised when molecular evidence pointed to a sister group relationship between the tribe Ceroxyleae and the phytelephantoid palms. The latter comprises three genera of morphological aberrant palms that have previously been considered a subfamily of their own. Here we present the results of a detailed study of the floral structure and development of the wax palm, Ceroxylon ceriferum, which aims at revealing derived traits shared by the sister tribes Ceroxyleae and Phytelepheae. A series of floral stages were sampled from Ceroxylon ceriferum growing in the central coastal range of Venezuela. The samples were prepared for scanning electronic microscopy and serial anatomical sectioning. The development of male and female flowers of Ceroxylon ceriferum was similar. The receptacle elongated early in the ontogeny. The perianth was differentiated into distinct sepals and petals and was characterized by a lack of postgenital fusion. The stamens were incepted centripetally in 2(-3) whorls. The outer whorl of three stamens was antesepalous. The inner whorl consisted of six stamens arranged in three antepetalous pairs. The flowers of Ceroxylon ceriferum share a lack of postgenital fusion in the perianth with members of the tribe Phytelepheae. The elongation of the receptacle is reminiscent of the receptacle expansion in Phytelepheae. However, the multistaminate condition in C. ceriferum is less extreme than in the Phytelepheae, and the stamen initiation is centripetal as opposed to centrifugal in the latter.

Sexual dimorphic floral development in dioecious plants revealed by transcriptome, phytohormone, and DNA methylation analysis in Populus tomentosa

Dioecious plants have evolved sex-specific floral development mechanisms. However, the precise gene expression patterns in dioecious plant flower development remain unclear. Here, we used andromonoecious poplar, an exceptional model system, to eliminate the confounding effects of genetic background of dioecious plants. Comparative transcriptome and physiological analysis allowed us to characterize sex-specific development of female and male flowers. Transcriptome analysis identified genes significantly differentially expressed between the sexes, including genes related to floral development, phytohormone synthesis and metabolism, and DNA methylation. Correlation analysis revealed a significant correlation between phytohormone signaling and gene expression, identifying specific phytohormone-responsive genes and their cis-regulatory elements. Two genes related to DNA methylation, METHYLTRANSFERASE1 (MET1) and DECREASED DNA METHYLATION 1 (DDM1), which are located in the sex determination region of Chromosome XIX, have differential expression between female and male flowers. A time-course analysis revealed that MET1 and DDM1 expression may produce different DNA methylation levels in female and male flowers. Understanding the interactions of phytohormone signaling, DNA methylation and target gene expression should lead to a better understanding of sexual differences in floral development. Thus, this study identifies a set of candidate genes for further studies of poplar sexual dimorphism and relates sex-specific floral development to physiological and epigenetic changes.

Sexual Dimorphism Floral MicroRNA Profiling and Target Gene Expression in Andromonoecious Poplar (Populus tomentosa)

Although the molecular basis of poplar sex-specific flower development remains largely unknown, increasing evidence indicates an essential role for microRNAs (miRNAs). The specific miRNA types and precise miRNA expression patterns in dioecious plant flower development remain unclear. Here, we used andromonoecious poplar, an exceptional model system, to eliminate the confounding effects of genetic background of dioecious plants. This system, combined with high-throughput sequencing and computational analysis, allowed us to characterize sex-specific miRNAomes from female and male flowers. Comparative miRNAome analysis combined with quantitative real-time PCR revealed the expression patterns of 27 miRNAs in poplar flower and showed that the targets of these miRNAs are involved in flower organogenesis, Ca2+ transport, phytohormone synthesis and metabolism, and DNA methylation. This paper describes a complex regulatory network consisting of these miRNAs expressed in sex-specific flower development in a dioecious plant. The conserved and novel miRNA locations were annotated in the Populus trichocarpa genome. Among these, miRNA Pto-F70 and 4 targets are located in the sex-determination regions of chromosome XIX. Furthermore, two novel miRNAs, Pto-F47 and Pto-F68, were shown for the first time to be regulatory factors in phytohormone interactions. To our knowledge, this report is the first systematic investigation of sex-specific flower-related miRNAs and their targets in poplar, and it deepens our understanding of the important regulatory functions of miRNAs in female and male flower development in this dioecious plant.

Effects of Low-light Stress on Male and Female Flower Development and Pollination and Fructification Ability of Different Maize ( Zea mays L.) Genotypes

A split plot experiment was conducted under field conditions to study the effects of low-light stress on the ear and tassel development,pollination and seed-setting rate using two maize(Zea mays L.)hybrids(low light sensitive hybrid Yuyu 22 and low light tolerant hybrid Zhengdan 958).Low-light stress from three days before tasselling to ten days after silking was obtained using50% transmittance shading nets.The results showed that low-light stress caused a slowdown of ear and tassel growth and development rate,delayed the tasselling date,silking date and full flowering date,prolonged the anthesis-silking interval(ASI).Daily pollen quantity was higher under low-light stress than that of the control at silking with a larger difference compared with the low light sensitive hybrid.For the low light sensitive hybrid under low light stress,reticula and gaps in the surface of pollen were increased,pollen aperture and its near part were severely deformed with significantly retraction,nutrient supply was reduced due to the reduction of starch grains in the pollen.While for the low light tolerant hybrid under low light stress,pollen surface characters were similar to that of control,with a little retraction of the pollen aperture and decrease of starch grain density in the pollen.Under low light stress,pollen activity,pollen emergence rate and pollen tube growth rate of the low light sensitive hybrid decreased while that of the low light tolerant hybrid increased.Silk elongation rate and number of silks emerged from the husks were decreased and decreased more in the ear tip than in the ear base.Ear length and kernel number per ear decreased in both hybrids and decreased more in low light sensitive hybrid.Prolonged ASI,deformed pollen,reduction of nutrient supply in pollen,decreased silk elongation rate and number of silks emerged from the husks,and the decrease of kernel IAA content and the increase of kernel ABA content were the primary causes leading to the reduction of ear kernel number.

In vitro pollen germination and pollen tube growth differences among Quercus robur L. clones in response to meteorological conditions

The impact of meteorological conditions on in vitro pollen germination and pollen tube growth during the initial phases of the development of male flowers in the Pedunculate Oak, Quercus robur, is studied. Phenological observations of male flowers and pollen sampling were performed on the field trial established with grafted Pedunculate Oak clones. During the investigation, weather conditions (absolute minimum and maximum daily air temperature, minimum absolute relative humidity of air and amount of precipitation) were recorded by an automatic meteorological station installed at the field trial. Influence of meteorological conditions on pollen germination and pollen tube growth was studied in the following stages of male flower: (I) during the last ten days of flower bud dormancy, (II) during swelling of the buds, (III) during bud burst and beginning of male catkins elongation, (IV) during the final stage of male flower catkins elongation. High temperatures and low relative air humidity during the bud burst and beginning of the male catkins elongation reduced pollen germination and pollen tube growth. Weather conditions did not significantly affect pollen germination and pollen tube growth during the swelling of flower buds, or in the final stage of male catkins elongation.

Structural Analysis of Reproductive Development in Staminate Flowers of Laurus nobilis L.

Male (staminat) flower development, being separated in 8 phases, was investigated in Laurus nobilis (Lauraceae) through the usage of histological sections and scanning electron microscopy (SEM) analysis. Flower development starts when apical meristem differentiates, followed by the conversion of this structure to floral meristem. Initial development phases comprise incidents similar to the ones of the female flower. 4 tepals and 8-10 stamens primordia develop through floral meristem in turn. In early stages of the development, sexual dimorphism occurs when the carpel primordium arrests. Filaments carry 2 nectaries in stamens which arise in 3 whorls. Anther wall consists of epidermis, endothecium, 2 or 3 middle layers and a single-layered glandular tapetum. Anthers are bisporangiate. Meiotic division is regular in pollen mother cells, and pollen grains do not contain aperture. Beside the pollen scattered individually within the pollen sacs, groups which contain some pollen tied to each other are rarely observed, as well. Pollen grains seldom germinate within microsporangium. Anthers are opened with 2 valves which widen from the base through the top. Accumulation of polysaccharides, lipids and proteins were identified by histochemical methods in stamens. These organic substances are greater within and around the vascular bundle compared to other tissues.

Stamen-derived bioactive gibberellin is essential for male flower development of Cucurbita maxima L.

Gibberellin (GA) signalling during pumpkin male flower development is highly regulated, including biosynthetic, perception, and transduction pathways. GA 20-oxidases, 3-oxidases, and 2-oxidases catalyse the final part of GA synthesis. Additionally, 7-oxidase initiates this part of the pathway in some cucurbits including Cucurbita maxima L. (pumpkin). Expression patterns for these GA-oxidase-encoding genes were examined by competitive reverse transcription-PCR (RT-PCR) and endogenous GA levels were determined during pumpkin male flower development. In young flowers, GA20ox3 transcript levels are high in stamens, followed by high levels of the GA precursor GA9. Later, just before flower opening, transcript levels for GA3ox3 and GA3ox4 increase in the hypanthium and stamens, respectively. In the stamen, following GA3ox4 expression, bioactive GA4 levels rise dramatically. Accordingly, catabolic GA2ox2 and GA2ox3 transcript levels are low in developing flowers, and increase in mature flowers. Putative GA receptor GID1b and DELLA repressor GAIPb transcript levels do not change in developing flowers, but increase sharply in mature flowers. Emasculation arrests floral development completely and leads to abscission of premature flowers. Application of GA4 (but not of its precursors GA12-aldehyde or GA9) restores normal growth of emasculated flowers. These results indicate that de novo GA4 synthesis in the stamen is under control of GA20ox3 and GA3ox4 genes just before the rapid flower growth phase. Stamen-derived bioactive GA is essential and sufficient for male flower development, including the petal and the pedicel growth.

Le déterminisme du sexe chez les cucurbitacées

Sex determination in plants leads to the development of unisexual flowers from an originally bisexual floral meristem. Cucurbits are not only species of agronomic interest but they also represent model species for the study of plant sex determination, because of their ability to harbor different sexual types. Such sexual forms are controlled by the identity of the alleles at the following loci: andromonoecious (a) and gynoecious (g) in melon, or androecious (a), Female (F), and Monoecious (M) in cucumber. We firstly showed that the andromonoecious a gene in melon encodes for an ACC synthase (CmACS7) and demonstrated that andromonoecy results from a mutation in the active site of the enzyme. Expression of the active enzyme inhibits the development of the male organs and is not required for carpel development. Because the a gene in melon and M gene in cucumber control the same sexual transition, monoecy to andromonoecy, we isolated the andromonoecy M gene in cucumber using a candidate gene approach in combination with genetic and biochemical analysis. We demonstrated the co-segregation of CsACS2, a close ortholog of CmACS7, with the M locus, and showed that the cucumber andromonoecious phenotype is also due to a loss of ACS enzymatic activity. CsACS2 is expressed specifically in carpel primordia of female flowers and should play a similar role to that of CmACS7 in melon in the inhibition of stamina development. Finally, we also showed that the transition from male to female flowers in the gynoecious lines results from epigenetic changes in the promoter of a C(2)H (2) zinc-finger transcription factor, CmWIP1. This epigenetic change is elicited by the insertion of a DNA transposon, which causes the spreading of DNA methylation to the CmWIP1 promoter. Expression of CmWIP1 leads to carpel abortion, resulting in the development of unisexual male flowers. From all these results, we built a model in which CmACS7 and CmWIP1 interact to control the development of male, female and hermaphrodite flowers in melon.

SCREENING OF FIFTEEN PUMPKIN (CUCURBITA MOSCHATA) GENOTYPES FOR YIELD AND SOME QUALITY UNDER SOHAG CONDITIONS

Eleven Egyptian landraces, two imported USA cultivars ('Etamp' and 'Butternut') and two Sudanese accessions (Khartoum and Omdurman) of pumpkin (Cucurbita moschata Duchesne) were assessed for yield and some fruit quality characteristics at the Experimental Farm, Faculty of Agriculture, Sohag University during two successive seasons, 2007 and 2008. There were significant differences among genotypes for all studied characters. 'Butternut' cultivar was the earliest in both female and male flower development but the lowest in fruit yield. Its fruits showed the least flesh thickness and the highest percent of total soluble solids (TSS). Khartoum accession was the latest to flower in both seasons. 'Etamp' cultivar exceeded all other genotypes in total fruit yield and fruit flesh thickness. Fruits of 'Bardis' Sohag accession had the lowest TSS in both seasons. The results of this study could be useful in breeding programs for improving pumpkin production in Upper Egypt.

Cloning and characterisation of two CTR1-like genes in Cucurbita pepo: regulation of their expression during male and female flower development

Ethylene is an essential regulator of flower development in Cucurbita pepo, controlling the sexual expression, and the differentiation and maturation of floral organs. To study the action mechanism of ethylene during the male and female flower development, we have identified two CTR1 homologues from C. pepo, CpCTR1 and CpCTR2, and analysed their expressions during female and male flower development and in response to external treatments with ethylene. CpCTR1 and CpCTR2 share a high homology with plant CTR1-like kinases, but differ from other related kinases such as the Arabidopsis EDR1 and the tomato LeCTR2. The C-terminal ends of both CpCTR1 and CpCTR2 have all the conserved motifs of Ser/Thr kinase domains, including the ATP-binding signature and the protein kinase active site consensus sequence, which suggests that CpCTR1 and CpCTR2 could have the same function as CTR1 in ethylene signalling. The transcripts of both genes were detected in different organs of the plant, including roots, leaves and shoots, but were mostly accumulated in mature flowers. During the development of male and female flowers, CpCTR1 and CpCTR2 expressions were concomitant with ethylene production, which indicates that both genes could be upregulated by ethylene, at least in flowers. Moreover, external treatments with ethylene, although did not alter the expression of these two genes in seedlings and leaves, were able to upregulate their expression in flowers. In the earlier stages of flower development, when ethylene production is very low, the expression of CpCTR1 and CpCTR2 is higher in male floral organs, which agrees with the role of these genes as negative regulators of ethylene signalling, and explain the lower ethylene sensitivity of male flowers in comparison with female flowers. The function of the upregulation of these two genes in later stages of female flower development, when the production of ethylene is also increased, is discussed.

A Fungal Parasite Regulates a Putative Female-Suppressor Gene Homologous to Maize Tasselseed2 and Causes Induced Hermaphroditism in Male Buffalograss

Parasitically induced hermaphroditism is a fascinating illustration of floral sex organ modification; however, knowledge of how parasites induce hermaphroditism in plants is limited. Here, we show the fungal parasite pistil smut induces development of female sex organs (pistils) in flowers of male buffalograss, potentially by downregulating a putative female-suppressor gene, BdTs2, homologous to maize Tasselseed2 (ZmTs2). Full-length BdTs2, isolated using rapid amplification of cDNA ends, exhibits 89% nucleotide sequence similarity with ZmTs2 and 85% amino acid sequence homology with ZmTs2 protein. Scanning electron micrographs demonstrate that unisexual buffalograss flowers develop through a process of selective abortion of opposite sex organs within hermaphroditic floral primordia. Quantitative real-time polymerase chain reaction showed that high expression levels of BdTs2 within male inflorescences correlate with the selective abortion of gynoecium, leading to the development of unisexual male flowers. RNA in situ hybridization confirmed the expression of BdTs2 precisely within vestigial gynoeciums of male flowers and not in other floral organs of the inflorescence. Furthermore, we show that BdTs2 expression is downregulated by pistil smut infection, which corresponds to the presence of pistils in flowers otherwise destined to become unisexual male. This study provides a potential molecular basis for pistil smut-induced hermaphroditism in male buffalograss.

A transposon-induced epigenetic change leads to sex determination in melon

Sex determination in plants leads to the development of unisexual flowers from an originally bisexual floral meristem. This mechanism results in the enhancement of outcrossing and promotes genetic variability, the consequences of which are advantageous to the evolution of a species. In melon, sexual forms are controlled by identity of the alleles at the andromonoecious (a) and gynoecious (g) loci. We previously showed that the a gene encodes an ethylene biosynthesis enzyme, CmACS-7, that represses stamen development in female flowers. Here we show that the transition from male to female flowers in gynoecious lines results from epigenetic changes in the promoter of a transcription factor, CmWIP1. This natural and heritable epigenetic change resulted from the insertion of a transposon, which is required for initiation and maintenance of the spreading of DNA methylation to the CmWIP1 promoter. Expression of CmWIP1 leads to carpel abortion, resulting in the development of unisexual male flowers. Moreover, we show that CmWIP1 indirectly represses the expression of the andromonoecious gene, CmACS-7, to allow stamen development. Together our data indicate a model in which CmACS-7 and CmWIP1 interact to control the development of male, female and hermaphrodite flowers in melon.

Floral Structure and Organization in Platanaceae

Developing and mature inflorescences and flowers of several representatives of Platanus were studied to clarify various aspects of floral structure and organization. Special attention was given to perianth differentiation. Extant Platanaceae are monoecious with unisexual flowers aggregated into compact, spherical inflorescence heads. Development of hairs in a basipetal direction subdivides the undifferentiated inflorescence surface into floral zones. Development of both male and female flowers of Platanus × hispanica begins with the initiation of a perianth whorl. Thereafter, the reproductive organs emerge on the floral apex: stamens in male flowers, staminodes and carpels in female flowers. The last organs to appear in both sexes are the small organs located between perianth and androecium. At anthesis, in both male and female flowers, organs of the first whorl are inconspicuous, scalelike, and only two to three cell layers thick. Alternating with these first thin organs is a whorl of short but fleshy organs. These second‐whorl organs are basally united with the stamens, forming a short androecial tube. They also show some structural similarities with stamens. These features support the hypothesis that the second‐whorl organs are of androecial (staminodial) origin. This hypothesis is further supported by the fossil record, where, in some taxa, second‐whorl organs are particularly similar to stamens, as well as by morphological comparisons with flowers of Proteaceae.

Tasselseed1 Is a Lipoxygenase Affecting Jasmonic Acid Signaling in Sex Determination of Maize

Sex determination in maize is controlled by a developmental cascade leading to the formation of unisexual florets derived from an initially bisexual floral meristem. Abortion of pistil primordia in staminate florets is controlled by a tasselseed-mediated cell death process. We positionally cloned and characterized the function of the sex determination gene tasselseed1 (ts1). The TS1 protein encodes a plastid-targeted lipoxygenase with predicted 13-lipoxygenase specificity, which suggests that TS1 may be involved in the biosynthesis of the plant hormone jasmonic acid. In the absence of a functional ts1 gene, lipoxygenase activity was missing and endogenous jasmonic acid concentrations were reduced in developing inflorescences. Application of jasmonic acid to developing inflorescences rescued stamen development in mutant ts1 and ts2 inflorescences, revealing a role for jasmonic acid in male flower development in maize.

ACC Synthase Genes are Polymorphic in Watermelon (Citrullus spp.) and Differentially Expressed in Flowers and in Response to Auxin and Gibberellin

The flowering pattern of watermelon species (Citrullus spp.) is either monoecious or andromonoecious. Ethylene is known to play a critical role in floral sex determination of cucurbit species. In contrast to its feminizing effect in cucumber and melon, in watermelon ethylene promotes male flower development. In cucumber, the rate-limiting enzyme of ethylene biosynthesis, 1-aminocyclopropane-1-carboxylate (ACC) synthase (ACS), regulates unisexual flower development. To investigate the role of ethylene in flower development, we isolated four genomic sequences of ACS from watermelon (CitACS1-4). Both CitACS1 and CitACS3 are expressed in floral tissue. CitACS1 is also expressed in vegetative tissue and it may be involved in cell growth processes. Expression of CitACS1 is up-regulated by exogenous treatment with auxin, gibberellin or ACC, the immediate precursor of ethylene. No discernible differential floral sex-dependent expression pattern was observed for this gene. The CitACS3 gene is expressed in open flowers and in young staminate floral buds (male or hermaphrodite), but not in female flowers. CitACS3 is also up-regulated by ACC, and is likely to be involved in ethylene-regulated anther development. The expression of CitACS2 was not detected in vegetative or reproductive organs but was up-regulated by auxin. CitACS4 transcript was not detected under our experimental conditions. Restriction fragment length polymorphism (RFLP) and sequence tagged site (STS) marker analyses of the CitACS genes showed polymorphism among and within the different Citrullus groups, including watermelon cultivars, Citrullus lanatus var. lanatus, the central subspecies Citrullus lanatus var. citroides, and the desert species Citrullus colocynthis (L).

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.

Floral structure and development in the dioecious Australian endemic Lomandra longifolia (Lomandraceae)

The micromorphology and histology of the development of male and female flowers of the dioecious Australian endemic species Lomandra longifolia Labill. was studied by means of scanning electron microscopy and light microscopy of entire and sectioned material. Although mature flowers are functionally unisexual, in the early stages of development pistillate and staminate flowers are identical and apparently bisexual. In a sequential fashion, six perianth parts are initiated within two alternating whorls, the sepals first and the petals second; six stamens are initiated in two alternating whorls of three stamens each, the first opposite the sepals and the second opposite the petals; and last, a central gynoecium is initiated. Following initiation, the two flower types diverge developmentally when the stamens become bilobed. In male flowers, cytological analysis of the slowly growing abortive pistil shows that megasporogenesis does not occur. Pistil abortion happens before meiosis whereas the stamens continue to develop until maturity and dehiscence. In female flowers, stamen arrest occurs before the onset of meiosis in microspore mother cells, after which the pistil continues its development through megasporogenesis and megagametogenesis. In all, 14 stages of floral development of both male and female flowers have been designated. Stages 1–6 of the two flower types were common to both sexes.