Floral Maturation Research Articles

Histochemical and molecular analyses reveal an insight into the scent volatiles synthesis and emission in ephemeral flowers of Murraya paniculata (L.) Jack.

Temporal histolocalization of floral volatiles in the petal epidermis of Murraya paniculata was found to be linked with the coordinated expression of candidate genes and successive accumulation of an internal pool of volatiles. Murraya paniculata (Rutaceae) is known for its highly fragrant ephemeral flowers that emit volatiles to attract nocturnal pollinators. To unfold the patterns of volatile emission in relation to floral life-span, we studied time-course accumulation and emission rate of scent volatiles at six timepoints of floral maturation, at an interval of 4h starting from the bud stage to the senescence stage on the next day. This study revealed the maximum emission rate of scent volatiles at the anthesis stage at 18:00h. This finding correlates well with the maximum accumulation of volatiles in the internal pool of the flowers at this stage. The key volatiles detected in both emitted and internal pools were benzaldehyde, benzeneacetaldehyde, linalool, caryophyllene, germacrene-D and α-farnesene. In addition, the internal pool also contained substantial amounts of indole, scopoletin, caffeine and osthole. To histochemically localize the temporal accumulation of major volatile groups in the epidermal cells, petal cross sections were stained with NaDi and ferric chloride to visualize terpenes and phenolics, respectively, under light microscope. Histolocalization studies showed a higher accumulation of terpenes at 14:00h and 18:00h, which subsequently was reduced as senescence approached. Significant phenolics in the abaxial and adaxial layers of the petal epidermis accumulated at 18:00h and at the early senescence (06:00h) stages. Furthermore, temporal localization of active shikimate dehydrogenase (SKDH) protein through in-gel activity assay demonstrated higher enzymatic activities at anthesis (18:00h) and fully bloomed (02:00h) stages, supporting the findings of higher accumulation of phenolic volatiles at 18:00h and 06:00h stages. Expression analysis of major candidate genes of floral scent volatiles pathway supported the hypothesis that the emission rate of floral fragrance reached its maximum at the anthesis (18:00h) stage. In contrast, biosynthesis of scent compounds started at the bud (14:00h) stage itself as indicated by the RT-PCR semi-quantitative estimation. As flowers of M. paniculata attract multiple pollinator species, this study could also serve as a springboard for pollination biology in Rutaceae, which includes important fruit crops.

Effect of High-Tunnel and Open-Field Production on the Yield, Cannabinoids, and Volatile Profiles in Industrial Hemp (Cannabis sativa L.) Inflorescence.

This study discovered the impact of high-tunnel (i.e., unheated greenhouse) and open-field production on two industrial hemp cultivars (SB1 and CJ2) over their yield parameters, cannabinoid development, and volatile profiles. Development of neutral cannabinoids (CBD, THC, and CBC), acidic cannabinoids (CBDA, THCA, and CBCA), and total cannabinoids during floral maturation were investigated. The volatile profiles of hemp flowers were holistically compared via HS-SPME-GC/MS. Findings indicated a high tunnel as an efficient practice for achieving greater total weight, stem number, and caliper, especially in the SB1 cultivar. Harvesting high-tunnel-grown SB1 cultivars during early flower maturation could obtain a high CBD yield while complying with THC regulations. Considering the volatile profiles, hemp flowers mainly consisted of mono- and sesquiterpenoids, as well as oxygenated mono- and sesquiterpenoids. Volatile analysis revealed the substantial impact of cultivars on the volatile profile compared to the production systems.

Influence of varieties of hemp, Cannabis sativa (Rosales: Cannabaceae), and fertilization rates on damage caused by corn earworm, Helicoverpa zea (Lepidoptera: Noctuidae).

Industrial hemp, Cannabis sativa L., production has been negatively impacted by larvae of corn earworm, Helicoverpa zea (Boddie), which feed on developing inflorescences. Adult H. zea oviposit on hemp once flowers develop, and late-instar larvae can cause serious loss to both quality and yield. A 2-year study to assess the influence of hemp variety and fertilization practices on damage caused by H. zea was conducted. Differences in damage ratings among varieties were observed in both years; however, the rate of nitrogen applied did not influence biomass yield or damage rating. These results indicate that increasing nitrogen fertility may not be an effective means of cultural control for mitigating damage from H. zea. Floral maturity was very influential on damage caused by H. zea as late-maturing varieties had much less floral injury than those which matured early in outdoor field trials. Some cannabinoids were also correlated to damage rating, but this relationship was due to late-maturing plants with immature flowers low in cannabinoid concentrations receiving less floral injury. Based on these results, the selection of high-yielding varieties that flower when ovipositional activity of H. zea is expected to decline should be the first step in an integrated pest management program for hemp production. This research expanded our knowledge of the role of fertility rate, varietal characteristics, cannabinoid profile, and floral maturity on damage caused by H. zea to hemp. Findings from this research will allow growers to make more informed agronomic decisions before planting to improve hemp production.

Hemp Morphology and Physiology Standards for Research and Industry Applications

Hemp (Cannabis sativa L.) research and commercial production has recently experienced a global revival motivated by passage of laws reversing long-standing prohibitions and by expansion in markets. Collaborative research has been initiated in response to renewed interest in hemp production, such as the American Society for Horticultural Science Hemp Research and Extension Professional Interest Group (ASHS Hemp). Collaborators new to this crop have identified a lack of standard definitions, descriptions, and procedures for cohesive study specific to hemp production. Standards are necessary for synthesis of data gathered across research and industry programs. ASHS Hemp convened a workshop of hemp researchers and industry representatives to establish consensus on a minimum set of standards for research data and industry assessments. The resulting morphology and physiology standards developed at the workshop are presented here with a focus on plant height, flowering time, and crop quality. Plant height was defined as the vertical distance between the root crown at the soil surface and the stem node (or tip) of the apical meristem of the tallest branch. Plant height was importantly distinguished from stem length and canopy height, which may differ based on pruning and management of the plant. Flowering time was defined to indicate date of initiation of inflorescence development as the earliest day terminal flowering clusters appear visually. Flowering time was distinguished from solitary flowering behavior and floral maturity. Crop quality was determined to be a feature that should be established first by industry based on market standards and then subsequently adopted by researchers targeting outcomes in specific areas. A standard moisture content for dry flower, seed, and straw must be established. A moisture content of 10% to 12% was identified as a current standard for floral yield, whereas 8% was identified as a moisture content standard for seed crops. Bast-to-hurd ratio and decortication efficiency were fiber quality metrics identified for minimum standards, and thousand seed weight, protein content, oil content, and oil composition were considered for minimum seed quality standards. The hemp research community is well positioned to standardize genomic references and establish best management practices for production targets. These efforts would be assisted by the adoption of the proposed standard definitions, descriptions, and procedures decided by consensus at the ASHS Hemp 2022 workshop.

Comparative transcriptome analysis reveals an insight into the candidate genes involved in anthocyanin and scent volatiles biosynthesis in colour changing flowers of Combretum indicum.

Combretum indicum is a widely cultivated ornamental species displaying the distinct phenomenon of floral colour change. Flowers display a gradual colour change from white to red, attributed to increased cyanidin 3-O glucoside in petal tissues. The differently coloured flowers also emanate a complex blend of VOCs with trans-linalool oxide (furanoid) as the major compound in the emission profile. To understand molecular mechanisms regulating floral colour shifts and scent biosynthesis, we performed Illumina transcriptome sequencing, including de novo assembly and functional annotation, for the two stages of floral maturation (white and red). Homology analysis with functional classification identified 84 and 42 candidate genes associated with pigment and scent biosynthesis, respectively. Genes encoding transcription factors, such as MYB, ERF, WD40, WRKY, NAC, bHLH and bZIP, that play critical roles in regulating specialized metabolism were also identified in the transcriptome data. Differences in expression of genes were consistent with accumulation patterns of anthocyanins in the two different flower colours. A clear upregulation of flavonoid biosynthesis genes in red flower tissue is associated with increased pigment content. RT-qPCR-based expression analyses gave results consistent with the RNA-Seq data, suggesting the sequencing data are consistent and reliable. This study presents the first report of genetic information for C. indicum. Gene sequences generated from RNA-Seq, along with candidate genes identified by pathway mapping and their expression profiles, provide a valuable resource for subsequent studies towards molecular understanding of specialized metabolism in C. indicum flowers.

Limited effect of environmental stress on cannabinoid profiles in high‐cannabidiol hemp (Cannabis sativa L.)

AbstractHemp (Cannabis sativa) is a burgeoning crop, but research‐based information about genetic and environmental effects of cannabinoid production is limited and will be essential for expanded cultivation. There are limited data available about the effect of environmental stressors on cannabinoid content, particularly for tetrahydrocannabinol (THC) in high‐cannabidiol (CBD) hemp. To address this, five stress treatments were applied in a replicated field trial with three high‐CBD hemp cultivars and cannabinoid content was assayed over a 3‐week time‐course spanning floral maturation. Cannabinoid production in terminal inflorescence shoot tip samples of three cultivars was measured under stress imposed by flooding, ethephon, powdery mildew, herbicide, and physical wounding in a split plot design. The treatments had limited effects on cannabinoid levels, with the exception of herbicide treatment which resulted in decreased cannabinoid content. Notably, there was no evidence that any of these stresses caused THC concentration or the ratio of THC to CBD to increase at harvest.

Season‐long characterization of high‐cannabinoid hemp (Cannabis sativa L.) reveals variation in cannabinoid accumulation, flowering time, and disease resistance

AbstractGiven the dramatic rise in high‐cannabinoid hemp (Cannabis sativa L.) production in the last decade, there is an increasingly urgent need to characterize available germplasm and develop knowledge to accelerate the breeding of uniform and stable cultivars. Despite persistent cultivation of hemp cultivars for grain and or fiber around the world, the diversity and genetic underpinning of key traits for breeding and cultivation are poorly understood. For 30 high‐cannabidiol hemp cultivars replicated on two field sites, we sought to evaluate yield, agronomic performance, and disease resistance while also conducting a detailed study of cannabinoid accumulation over the course of floral maturation. We observed significant variation in both within and among cultivars. During the growing season, the plants clustered into five groups by growth rate and varied in flowering time from photoperiod insensitive to photoperiod sensitive with very short critical photoperiods. Based on the observed ratio of total potential cannabidiol (CBD) to total potential tetrahydrocannabinol (THC), there was segregation for cannabinoid chemotype in some seeded cultivar populations. Analysis of cannabichromene (CBC) production revealed that some cultivars had a discretely lower CBD:CBC ratio than the others. There was a continuous range of powdery mildew severity by cultivar, with one that had little to no observed powdery mildew suggesting it might have genetic resistance. Biomass production at harvest was strongly influenced by location and cultivar, and there was variation by cultivar in the relative cannabinoid production in shoot tip samples compared with whole plant samples. While our results provide preliminary guidance regarding relative performance of current cultivars, our analyses indicate a need for additional hemp breeding to provide stable, uniform, and legally compliant cultivars with improved disease resistance and flowering times optimized for the latitudes of different growing locations.

Diversification of petal monoterpene profiles during floral development and senescence in wild roses: relationships among geraniol content, petal colour, and floral lifespan

Wild roses store and emit a large array of fragrant monoterpenes from their petals. Maximisation of fragrance coincides with floral maturation in many angiosperms, which enhances pollination efficiency, reduces floral predation, and improves plant fitness. We hypothesized that petal monoterpenes serve additional lifelong functions such as limiting metabolic damage from reactive oxygen species (ROS), and altering isoprenoid hormonal abundance to increase floral lifespan. Petal monoterpenes were quantified at three floral life-stages (unopened bud, open mature, and senescent) in 57 rose species and 16 subspecies originating from Asia, America, and Europe, and relationships among monoterpene richness, petal colour, ROS, hormones, and floral lifespan were analysed within a phylogenetic context. Three distinct types of petal monoterpene profiles, revealing significant developmental and functional differences, were identified: Type A, species where monoterpene abundance peaked in open mature flowers depleting thereafter; Type B, where monoterpenes peaked in senescing flowers increasing from bud stage, and a rare Type C (8 species) where monoterpenes depleted from bud stage to senescence. Cyclic monoterpenes peaked during early floral development, whereas acyclic monoterpenes (dominated by geraniol and its derivatives, often 100-fold more abundant than other monoterpenes) peaked during floral maturation in Type A and B roses. Early-diverging roses were geraniol-poor (often Type C) and white-petalled. Lifetime changes in hydrogen peroxide (H2O2) revealed a significant negative regression with the levels of petal geraniol at all floral life-stages. Geraniol-poor Type C roses also showed higher cytokinins (in buds) and abscisic acid (in mature petals), and significantly shorter floral lifespan compared with geraniol-rich Type A and B roses. We conclude that geraniol enrichment, intensification of petal colour, and lower potential for H2O2-related oxidative damage characterise and likely contribute to longer floral lifespan in monoterpene-rich wild roses.

Gene expression trajectories during male and female reproductive development in balsam poplar (Populus balsamifera L.)

Plant reproductive development from the first appearance of reproductively committed axes through to floral maturation requires massive and rapid remarshalling of gene expression. In dioecious species such as poplar this is further complicated by divergent male and female developmental programs. We used seven time points in male and female balsam poplar (Populus balsamifera L.) buds and catkins representing the full annual flowering cycle, to elucidate the effects of time and sex on gene expression during reproductive development. Time (developmental stage) is dominant in patterning gene expression with the effect of sex nested within this. Here, we find (1) evidence for five successive waves of alterations to the chromatin landscape which may be important in setting the overall reproductive trajectory, regardless of sex. (2) Each individual developmental stage is further characterized by marked sex-differential gene expression. (3) Consistent sexually differentiated gene expression regardless of developmental stage reveal candidates for high-level regulators of sex and include the female-specific poplar ARR17 homologue. There is also consistent male-biased expression of the MADS-box genes PISTILLATA and APETALA3. Our work provides insights into expression trajectories shaping reproductive development, its potential underlying mechanisms, and sex-specific translation of the genome information into reproductive structures in balsam poplar.

Carbohydrate Metabolism and Signaling in Squash Nectaries and Nectar Throughout Floral Maturation.

Floral nectar is a sugary solution produced by plants to entice pollinator visitation. A general mechanism for nectar secretion has been established from genetic studies in Arabidopsis (Arabidopsis thaliana); however, supporting metabolic and biochemical evidence for this model is scarce in other plant species. We used squash (Cucurbita pepo) to test whether the genetic model of nectar secretion in Arabidopsis is supported at the metabolic level in other species. As such, we analyzed the expression and activity of key enzymes involved in carbohydrate metabolism in squash nectaries throughout floral maturation and the associated starch and soluble sugars, as well as nectar volume and sugar under different growth conditions. Here we show that the steps that are important for nectar secretion in Arabidopsis, including nectary starch degradation, Suc synthesis, and Suc export, are supported by metabolic and biochemical data in C. pepo Additionally, our findings suggest that sugars imported from the phloem during nectar secretion, without prior storage as starch, are important for generating C. pepo nectar. Finally, we predict that trehalose and trehalose 6-P play important regulatory roles in nectary starch degradation and nectar secretion. These data improve our understanding of how nectar is produced in an agronomically relevant species with the potential for use as a model to help us gain insight into the biochemistry and metabolism of nectar secretion in flowering plants.

An integrated transcriptomics and metabolomics analysis of the Cucurbita pepo nectary implicates key modules of primary metabolism involved in nectar synthesis and secretion.

Nectar is the main reward that flowers offer to pollinators to entice repeated visitation. Cucurbita pepo (squash) is an excellent model for studying nectar biology, as it has large nectaries that produce large volumes of nectar relative to most other species. Squash is also monoecious, having both female and male flowers on the same plant, which allows comparative analyses of nectary function in one individual. Here, we report the nectary transcriptomes from both female and male nectaries at four stages of floral maturation. Analysis of these transcriptomes and subsequent confirmatory experiments revealed a metabolic progression in nectaries leading from starch synthesis to starch degradation and to sucrose biosynthesis. These results are consistent with previously published models of nectar secretion and also suggest how a sucrose‐rich nectar can be synthesized and secreted in the absence of active transport across the plasma membrane. Nontargeted metabolomic analyses of nectars also confidently identified 40 metabolites in both female and male nectars, with some displaying preferential accumulation in nectar of either male or female flowers. Cumulatively, this study identified gene targets for reverse genetics approaches to study nectary function, as well as previously unreported nectar metabolites that may function in plant‐biotic interactions.

Identification and Functional Characterization of Sugarcane Invertase Inhibitor (ShINH1): A Potential Candidate for Reducing Pre- and Post-harvest Loss of Sucrose in Sugarcane.

In sugarcane, invertase enzymes play a key role in sucrose accumulation and are also involved in futile reactions where sucrose is continuously degraded during the pre- and post-harvest period, thereby reducing sugar yield and recovery. Invertase inhibitor (INVINH) proteins play a key role in post-translation regulation of plant invertases through which sucrose hydrolysis is controlled. INVINH proteins are small (18 kDa) members of the pectin methylesterase inhibitor superfamily and they are moderately conserved across plants. In the present study, we identified two INVINH genes from sugarcane, ShINH1 and ShINH2. In silico characterization of the encoded proteins revealed 43% sequence identity at the amino acid level, confirming the non-allelic nature of the proteins. The presence of putative signal peptide and subcellular targeting sequences revealed that ShINH1 and ShINH2 likely have apoplasmic and vacuolar localization, respectively. Experimental visualization of ShINH1–GFP revealed that ShINHI is indeed exported to the apoplast. Differential tissue-specific and developmental expression of ShINH1 between leaf, stalk, flower and root suggest that it plays a role in controlling source-sink metabolic regulation during sucrose accumulation in sugarcane. ShINH1 is expressed at relatively high levels in leaves and stalk compared to flowers and roots, and expression decreases significantly toward internodal maturity during stalk development. ShINH1 is expressed at variable levels in flowers with no specific association to floral maturity. Production of recombinant ShINH1 enabled experimental validation of protein function under in vitro conditions. Recombinant ShINH1 potently inhibited acid invertase (IC50 22.5 nM), making it a candidate for controlling pre- and post-harvest deterioration of sucrose in sugarcane. Our results indicate that ShINH1 and ShINH2 are likely to play a regulatory role in sucrose accumulation and contribute to the improvement of sugar yield and recovery in sugarcane.

Phalaenopsis flowering locus VE regulates floral organ maturation.

PaFVE is low ambient temperature-inducible and acts as a systemic regulator in the early stage of floral development in Phalaenopsis. Phalaenopsis aphrodite: subsp. formosana, a native orchid species of Taiwan, is an economically important ornamental crop that requires low ambient temperature for floral transition. Currently, limited genetic information about such orchid species hampers genetic manipulation for specific or improved floral traits, and the control of flowering time independent of temperature regulation. In this study, the sequence of the full-length of Phalaenopsis flowering locus VE (PaFVE) gene was determined. Spatial and temporal expression studies showed that mRNA transcripts of PaFVE were inducible by low ambient temperature, and high levels of expression occurred after spiking initiation and remained high throughout the early stage of floral development. Further investigation revealed that floral organ development was impeded in PaFVE-silenced P. aphrodite, but flowering time and floral organogenesis were not compromised. Analysis of the downstream flowering genes suggested that the delay in floral maturation is associated with a corresponding decrease in the expression of downstream flowering genes, PaSOC1, PaSOC1L and PaAGL24. The ectopic expression of PaFVE in Arabidopsis resulted in an accelerated flowering time, accompanied by an increase in the expression of AtSOC1, thus revealing the functional role of PaFVE as a floral regulator. Overall, our results demonstrate that PaFVE has evolutionarily diverged and conserved functions, and serves as a regulator of floral organ maturation in Phalaenopsis and a regulator of flowering time in Arabidopsis.

Behind the diversity: Ontogenies of radiate, disciform, and discoid capitula of Chrysanthemum and its allies

AbstractAsteraceae are distinguishable by their head‐like inflorescence called a capitulum. There are a variety of capitulum types due to different combinations of different composing florets. Morphological evolution of capitula must involve homoplasy under selective constraints. Therefore, a comparative examination of ontogenies of divergent capitula among phylogenetically close taxa is important to investigate the genetic and molecular bases of this trait. In the subtribe Artemisiinae, tribe Anthemideae, three closely related genera have different flower heads: Chrysanthemum with the radiate, Ajania with the disciform, and Stilpnolepis with the discoid capitula. Through observations using scanning electron microscopy, we examined the morphogenetic processes of flower heads of representative species of these genera. The morphological differentiation of the discoid from the radiate/disciform capitula occurred early in the floral initiation stage. The development of the ray/marginal florets of the radiate/disciform capitula lagged behind the outermost disc florets throughout the phases of floral initiation and floral organogenesis. Primordia of their 1–2 dorsal corolla lobes and all stamens ceased to grow soon after initiation. In contrast, all florets on a discoid capitulum developed equally in rates and modes into bisexual actinomorphic flowers. Differentiation between the radiate and the disciform capitula occurred late during the floral maturation stage. Our data provided evidence to the interpretation that uniformly acropetal development only occurs in species with homogamous flower heads, and to the idea that the timing of ontogenetic divergence between taxa is positively correlated with the phylogenetic distance of the taxa.

Morphometric analysis of floral variation in thePyrola pictaspecies complex (Ericaceae): interpretation and implications for ecological and phylogenetic differentiation

The Pyrola picta species complex of western North America comprises four species (P. picta, P. dentata, P. aphylla and P. crypta) that grow sympatrically in some parts of their collective ranges, have remarkably similar flowers and share pollinators. These species do not exhibit the genetic signatures typical of random or heterospecific mating, but instead show genetic divergence patterns indicating that they maintain surprising levels of reproductive isolation. To better understand how species boundaries are maintained, the current study uses statistical ordination analyses to determine whether species isolation across shared geographical ranges might be achieved through subtle differences in floral characters among species. The possible contribution of differences in flowering phenology (e.g. temporal reproductive isolation) to reproductive isolation was also evaluated for the small subset of populations in which two or more species occur in direct sympatry. Among species in the P. picta complex, there are both phylogenetic and geographical trends in some floral characteristics, whereas other characters do not covary with either geography or species identity. In several sympatric populations, differences in flowering phenology among species suggest that timing plays a major role in non-random (i.e. mainly conspecific) mating. The conclusions of this study are that reproductive isolation in the P. picta species complex is reinforced by differences in the timing of floral maturation and the morphologies of androecium and floral display characters. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 00, 000–000.

Cryptic species in Pagamea coriacea sensu lato (Rubiaceae): evidence from morphology, ecology and reproductive behavior in a sympatric context

In this study we explore morphological and ecological variation in sympatric populations of Pagamea coriacea s.l. - a species complex from white-sand vegetation in the Amazon. A total of 147 trees were sampled and monitored at three nearby sites in Central Amazon, Brazil. Multivariate analyses of morphology indicated two distinct groups (A and B), which also differed in bark type, each containing subgroups associated with sexual dimorphism. However, a single hermaphroditic individual was observed within group B. As expected, all pistillate plants produced fruits, but 23% of the staminate plants of group B, and 5% of group A also produced fruits. This variation suggests that the sexual systems of both groups are between dioecy and gynodioecy. There was an overlap in flowering phases between the two groups, but the pattern of floral maturation differed. Ecologically, plants of group B were found in more shaded habitats and over sandstone bedrocks, while group A was prevalent in deeper sandy soils as canopy plants. The significances of morphological and environmental differences were tested by a multivariate analysis of variance, and a canonical discriminant analysis assessed the importance of variables. The coexistence in sympatry of two discrete morphological groups in the P. coriacea s.l., with different habitat preferences and reproductive behaviors, indicates they represent distinct species.

Boronia heterophylla vase life is influenced more by ethylene than by bacterial numbers or vase solution pH

Bacterial proliferation in both vase solutions and in cut flower stems has been implicated in reducing the vase life of numerous genera. Boronia heterophylla F. Muell. (Red Boronia) vase life was assessed at two stages of floral maturity for nine vase solution treatments covering a pH range of 2.5–5.7. Vase life for advanced harvest maturity stems ranged from 4.2 d in 10mM citric acid+50mgL−1 chlorine (pH 2.5) to 12.9 d after STS pulsing (pH 5.7). For normal harvest maturity stems, the corresponding range was 5.8–19.0 d, respectively. Vase solutions containing 50mgL−1 chlorine biocide resulted in decreased longevity. In contrast, pulsing with the ethylene-binding inhibitor, STS, significantly increased vase life. The number of bacteria in the vase solutions after 11 d was determined in stems of advanced maturity. The solution with the greatest number of bacteria, 4.0×1010cfumL−1, was water used after STS pulsing and in which the flowers lasted longest. Vase solution bacteria were enumerated on days 0, 3, 6, 9 and 12 of the vase period with stems of normal harvest maturity. There was no relationship between vase life and vase solution bacterial numbers (R¯2=0.000). Moreover, there was a negative relationship between numbers of bacteria in basal 0–5cm stem segments and vase life. As no correlations were evident between longevity and either the pH or vase solution bacterial numbers, B. heterophylla vase life was evidently limited principally by ethylene action.

The Anatomy of the Stigma and Style from Cyclamen persicum (Mill.) cv. “Pure White” and its Relation to Pollination Success

Old flowers of Cyclamen generate few or no seeds. To understand the pollination problems of Cyclamen we investigate the general anatomy of the stigma and the style of Cyclamen persicum by scanning electron microscopy at different stages of floral maturity. Our investigations confirm that there is a hollow style. Against data commonly found in the literature, we present evidence of pollen germination and tube growth that show the stigma is not outside the style but inside it. Furthermore the maturation process of the style during the flowering time indicates a mechanism by which the stigma becomes shut off through closure at the terminal aperture of the style. At 3 to 5 days after anthesis there was the beginning closure of the style which was nearly completed at 21 days. The substance which leads to the closure is still unknown. The closure of the hollow style is a probable cause for failure of seed set in flowers not pollinated early in anthesis.

Floral phenology and compatibility of sawgrass, Cladium jamaicense (Cyperaceae)

Sawgrass, Cladium jamaicense, is the dominant macrophyte in the Florida Everglades. We examined sawgrass flowering phenology and compatibility reactions in ex situ and in situ populations over 2 yr. Sawgrass flowers in May in southern Florida. Flower maturation was relatively synchronous within an inflorescence. Along the entire inflorescence, functionally male flowers emerged initially, followed by stigmas, then anthers of hermaphroditic flowers. Flowers of each sex expanded over 2 d with less than 1 d in between, totaling 6- 7 d for an inflorescence to complete flowering. Hand pollinations showed that sawgrass was self-compatible and not pollen-limited, because open pollinations produced fruit set similar to self- and cross-pollinations. Fruit set was low in autogamy and manipulation treatments. Manipulation treatments were used to study the effect of exposure to airborne pollen during hand pollinations. This treatment thus provides a useful technique for studies on the in situ compatibility of wind-pollinated graminoids. Sawgrass was able to self-fertilize, but the timing of flower maturation on an inflorescence promoted outcrossing. Actual outcrossing rates in sawgrass thus depend on clonal architecture and the timing of floral maturation on other inflorescences within a clone rather than on inflorescences of other genets in a population.

Glycosidically bound volatile components of Nicotiana sylvestris and N. Suaveolens flowers

Glycosidically bound volatile components were extracted from flowers of Nicotiana sylvestris and N. suaveolens and isolated using liquid column chromatography with an Amberlite XAD-2 resin. These glycosides, which were putative precursors of fragrance compounds, were hydrolysed enzymatically and a number of the volatiles released were subsequently identified by GC-MS including compounds not previously identified in the floral headspace of these species. Average yields of volatiles released from glycosides were ca 230 and 1050 μg g −1 for N. sylvestris and N. suaveolens flowers, respectively. Compounds in the glycosidically bound fraction were all phenylpropanoid-derived volatiles (e.g. benzyl alcohol, benzaldehyde, E-cinnamyl alcohol, benzyl salicylate) with the exception of the monoterpene α-terpineol. No pronounced diurnal changes in the levels of glycosidically bound volatile compounds could be noted from flowers of either species. However, pronounced differences were obtained in the concentrations of glycosidically bound volatiles at different floral maturity stages.