LEAFY, TERMINAL FLOWER1 and AGAMOUS are functionally conserved but do not regulate terminal flowering and floral determinacy in Impatiens balsamina

In Impatiens balsamina a lack of commitment of the meristem during floral development leads to the continuous requirement for a leaf-derived floral signal. In the absence of this signal the meristem reverts to leaf production. Current models for Arabidopsis state that LEAFY (LFY) is central to the integration of floral signals and regulates flowering partly via interactions with TERMINAL FLOWER1 (TFL1) and AGAMOUS (AG). Here we describe Impatiens homologues of LFY, TFL1 and AG (IbLFY, IbTFL1 and IbAG) that are highly conserved at a sequence level and demonstrate homologous functions when expressed ectopically in transgenic Arabidopsis. We relate the expression patterns of IbTFL1 and IbAG to the control of terminal flowering and floral determinacy in Impatiens. IbTFL1 is involved in controlling the phase of the axillary meristems and is expressed in axillary shoots and axillary meristems which produce inflorescences, but not in axillary flowers. It is not involved in maintaining the terminal meristem in either an inflorescence or indeterminate state. Terminal flowering in Impatiens appears therefore to be controlled by a pathway that uses a different integration system than that regulating the development of axillary flowers and branches. The pattern of ovule production in Impatiens requires the meristem to be maintained after the production of carpels. Consistent with this morphological feature IbAG appears to specify stamen and carpel identity, but is not sufficient to specify meristem determinacy in Impatiens.     

Reversion of flowering

Reversion from floral to vegetative growth is under environmental control in many plant species. However the factors regulating floral reversion, and the events at the shoot apex that take place when it occurs, have received less attention than those associated with the transition to flowering. Reversions may be categorized as flower reversion, in which the flower meristem resumes leaf production, or inflorescence reversions, in which the meristem ceases to initiate bracts with flowers in their axils and begins instead to make leaves with vegetative branches in their axils. Related to these two types of reversion, but distinct from them, are examples of partial flowering, when non-floral meristems grow out so that the plant begins to grow vegetatively again. Anomalous or proliferous flowers may form as a result of unfavourable growth conditions or viral infection, but these do not necessarily involve flower reversions.     

Non-reversion of Impatiens in the absence of meristem commitment

Purple-flowered plants of Impatiens balsamina maintained floral development on transfer from inductive short days (SD) to long days (LD), a treatment in which red-flowered plants of Impatiens are known to revert to leaf production. An investigation into the non-reverting nature of purple-flowered plants was carried out to establish whether these plants achieved meristem commitment or whether their non-reverting state was controlled by the leaves. When the leaves that had unfolded during the inductive SD treatment were removed at the time of transfer to LD, the purple-flowered plant did revert. This result suggests that, as in red-flowered Impatiens, meristem commitment is absent, but that purple-flowered plants maintain flowering in LD conditions because of a more permanent supply of signal from their leaves than occurs in red-flowered plants. A working hypothesis is proposed to explain how a signal from the leaves can retain a controlling role during flower development.Key words: Floral commitment, Impatiens, floral reversion, floricaula.      

ABERRANT PANICLE ORGANIZATION 2/RFL, the rice ortholog of Arabidopsis LEAFY, suppresses the transition from inflorescence meristem to floral meristem through interaction with APO1

The temporal and spatial control of meristem identity is a key element in plant development. To better understand the molecular mechanisms that regulate inflorescence and flower architecture, we characterized the rice aberrant panicle organization 2 (apo2) mutant which exhibits small panicles with reduced number of primary branches due to the precocious formation of spikelet meristems. The apo2 mutants also display a shortened plastochron in the vegetative phase, late flowering, aberrant floral organ identities and loss of floral meristem determinacy. Map-based cloning revealed that APO2 is identical to previously reported RFL gene, the rice ortholog of the Arabidopsis LEAFY (LFY) gene. Further analysis indicated that APO2/RFL and APO1, the rice ortholog of Arabidopsis UNUSUAL FLORAL ORGANS, act cooperatively to control inflorescence and flower development. The present study revealed functional differences between APO2/RFL and LFY. In particular, APO2/RFL and LFY act oppositely on inflorescence development. Therefore, the genetic mechanisms for controlling inflorescence architecture have evolutionarily diverged between rice (monocots) and Arabidopsis (eudicots).     

Competence to respond to floral inductive signals requires the homeobox genes PENNYWISE and POUND-FOOLISH

The transition from vegetative to reproductive development establishes new growth patterns required for flowering. This switch is controlled by environmental and/or intrinsic developmental cues that converge at the shoot apical meristem (SAM). During this developmental transition, floral inductive signals cause the vegetative meristem to undergo morphological changes that are essential for flowering. Arabidopsis plants containing null mutations in two paralogous BEL1-like (BELL) homeobox genes, PENNYWISE (PNY) and POUND-FOOLISH (PNF), disrupt the transition from vegetative to reproductive development. These double mutants are completely unable to flower even though the SAM displays morphological and molecular changes that are consistent with having received floral inductive signals. These studies establish a link between the competence to receive floral inductive signals and restructuring of the SAM during floral evocation.     

A leaf-derived signal is a quantitative determinant of floral form in Impatiens

The completion of flower development in Impatiens balsamina requires continuous inductive (short-day) conditions. We have previously shown that a leaf-derived signal has a role in floral maintenance. The research described here analyzes the role of the leaf in flower development. Leaf removal treatments, in which plants were restricted to a specified number of leaves, resulted in flowers with increased petal number, up to double that of the undefoliated control. Similar petal number increases (as well as changes in bract number or morphology) were recorded when plants began their inductive treatment at a late developmental age or when plants of a nonreverting line (capable of floral maintenance in the absence of continuous short days) were transferred from short days to long days. Our data imply that the increased petal number was neither a response to stress effects associated with leaf removal nor a result of alterations in primordium initiation rates or substitutions of petals for stamens. Rather, the petal initiation phase was prolonged when the amounts of a leaf-derived signal were limiting. We conclude that a leaf-derived signal has a continuous and quantitative role in flower development and propose a temporal model for the action of organ identity genes in Impatiens. This work adds a new dimension to the prevailing ABC model of flower development and may provide an explanation for the wide variety and instabilities of floral form seen among certain species in nature.     

Changes in Apical Growth and Phyllotaxis on Flowering and Reversion in Impatiens balsamina L.

When plants of Impatiens balsamina L were subjected to 5 shortdays and then re-placed in long days, they began to form a terminalflower and then reverted to vegetative growth at this terminalshoot apex The onset of flowering was accompanied by an increasein the rate of initiation of primordia, an increase in the growthrate of the apex, a change in primordium arrangement from spiralto whorled or pseudo-whorled, a lack of internodes, and a reductionm the size at initiation of the primordia and also of the stemfrusta which give rise to nodal and internodal tissues On reversion,parts intermediate between petals and leaves were formed, followedby leaves, although in reverted apices the size at initiationand the arrangement of primordia remained the same as in thefloweing apex The apical growth rate and the rate of primordiuminitiation were less in the reverted apices than in floral apicesbut remained higher than in the original vegetative apex Sincethe changes in apical growth which occur on the transition toflowering are not reversed on reversion, the development oforgans as leaves or petals is not directly related to the growthrate of the apex, or the arrangement, rate of initiation orsize at initiation of primordia Impatiens balsamina L, flower reversion, evocation, phyllotaxis, shoot meristem     

Expression of CENTRORADIALIS (CEN) and CEN-like genes in tobacco reveals a conserved mechanism controlling phase change in diverse species.

Plant species exhibit two primary forms of flowering architecture, namely, indeterminate and determinate. Antirrhinum is an indeterminate species in which shoots grow indefinitely and only generate flowers from their periphery. Tobacco is a determinate species in which shoot meristems terminate by converting to a flower. We show that tobacco is responsive to the CENTRORADIALIS (CEN) gene, which is required for indeterminate growth of the shoot meristem in Antirrhinum. Tobacco plants overexpressing CEN have an extended vegetative phase, delaying the switch to flowering. Therefore, CEN defines a conserved system controlling shoot meristem identity and plant architecture in diverse species. To understand the underlying basis for differences between determinate and indeterminate architectures, we isolated CEN-like genes from tobacco (CET genes). In tobacco, the CET genes most similar to CEN are not expressed in the main shoot meristem; their expression is restricted to vegetative axillary meristems. As vegetative meristems develop into flowering shoots, CET genes are downregulated as floral meristem identity genes are upregulated. Our results suggest a general model for tobacco, Antirrhinum, and Arabidopsis, whereby the complementary expression patterns of CEN-like genes and floral meristem identity genes underlie different plant architectures.     

Reproductive meristem fates in Gerbera

Flowering plants go through several phases between regular stem growth and the actual production of flower parts. The stepwise conversion of vegetative into inflorescence and floral meristems is usually unidirectional, but under certain environmental or genetic conditions, meristems can revert to an earlier developmental identity. Vegetative meristems are typically indeterminate, producing organs continuously, whereas flower meristems are determinate, shutting down their growth after reproductive organs are initiated. Inflorescence meristems can show either pattern. Flower and inflorescence development have been investigated in Gerbera hybrida, an ornamental plant in the sunflower family, Asteraceae. Unlike the common model species used to study flower development, Gerbera inflorescences bear a fixed number of flowers, and the architecture of the flowers differ in that Gerbera ovaries are inferior (borne below the perianth). This architectural difference has been exploited to show that floral meristem determinacy and identity are spatially and genetically distinct in Gerbera, and we have shown that a single SEPALLATA-like MADS domain factor controls both flower and inflorescence meristem fate in the plant. Although these phenomena have not been directly observed in Arabidopsis, the integrative role of the SEPALLATA function in reproductive meristem development may be general for all flowering plants.     

Cytokinin levels in leaves, leaf exudate and shoot apical meristem of Arabidopsis thaliana during floral transition

Understanding the complete picture of floral transition is still impaired by the fact that physiological studies mainly concern plant species whose genetics is poorly known, and vice versa. Arabidopsis thaliana has been successfully used to unravel signalling pathways by genetic and molecular approaches, but analyses are still required to determine the physiological signals involved in the control of floral transition. In this work, the putative role of cytokinins was investigated using vegetative plants of Arabidopsis (Columbia) induced to flower synchronously by a single 22 h long day. Cytokinins were analysed in leaf extracts, leaf phloem exudate and in the shoot apical meristem at different times during floral transition. It was found that, in both the leaf tissues and leaf exudate, isopentenyladenine forms of cytokinins increased from 16 h after the start of the long day. At 30 h, the shoot apical meristem of induced plants contained more isopentenyladenine and zeatin than vegetative controls. These cytokinin increases correlate well with the early events of floral transition.     

Reversion of flowering in Glycine Max (Fabaceae)

Photoperiodic changes, if occurring before a commitment to flowering is established, can alter the morphological pattern of plant development. In this study, Glycine max (L.) Merrill cv. Ransom plants were initially grown under an inductive short-day (SD) photoperiod to promote flower evocation and then transferred to a long-day (LD) photoperiod to delay flower development by reestablishing vegetative growth (SD-LD plants). Some plants were transferred back to SD after 4-LD exposures to repromote flowering (SD-LD-SD plants). Alterations in organ initiation patterns, from floral to vegetative and back to floral, are characteristic of a reversion phenomenon. Morphological features that occurred at the shoot apical meristem in SD, LD, SD-LD, and SD-LD-SD plants were observed using scanning electron microscopy (SEM). Reverted plants initiated floral bracts and resumed initiation of trifoliolate leaves in the two-fifths floral phyllotaxy prior to terminal inflorescence development. When these plants matured, leaf-bract intermediates were positioned on the main stem instead of trifoliolate leaves. Plants transferred back to a SD photoperiod flowered earlier than those left in LD conditions. Results indicated that in plants transferred between SDs and LDs, photoperiod can influence organ initiation in florally evoked, but not committed, G. max plants.     

Control of Arabidopsis flowering: the chill before the bloom

The timing of the floral transition has significant consequences for reproductive success in plants. Plants gauge both environmental and endogenous signals before switching to reproductive development. Many temperate species only flower after they have experienced a prolonged period of cold, a process known as vernalization, which aligns flowering with the favourable conditions of spring. Considerable progress has been made in understanding the molecular basis of vernalization in Arabidopsis. A central player in this process is FLC, which blocks flowering by inhibiting genes required to switch the meristem from vegetative to floral development. Recent data shows that many regulators of FLC alter chromatin structure or are involved in RNA processing.     

Characterization of tomato (Solanum lycopersicum L.) mutants affected in their flowering time and in the morphogenesis of their reproductive structure

The impact of the season on flowering time and the organization and morphogenesis of the reproductive structures are described in three tomato mutants: compound inflorescence (s), single flower truss (sft), and jointless (j), respectively, compared with their wild-type cultivars Ailsa Craig (AC), Platense (Pl), and Heinz (Hz). In all environmental conditions, the sft mutant flowered significantly later than its corresponding Pl cultivar while flowering time in j was only marginally, but consistently, delayed compared with Hz. The SFT gene and, to a lesser extent, the J gene thus appear to be constitutive flowering promoters. Flowering in s was delayed in winter but not in summer compared with the AC cultivar, suggesting the existence of an environmentally regulated pathway for the control of floral transition. The reproductive structure of tomato is a raceme-like inflorescence and genes regulating its morphogenesis may thus be divided into inflorescence and floral meristem identity genes as in Arabidopsis. The s mutant developed highly branched inflorescences bearing up to 200 flowers due to the conversion of floral meristems into inflorescence meristems. The S gene appears to be a floral meristem identity gene. Both sft and j mutants formed reproductive structures containing flowers and leaves and reverting to a vegetative sympodial growth. The SFT gene appears to regulate the identity of the inflorescence meristem of tomato and is also involved, along with the J gene, in the maintenance of this identity, preventing reversion to a vegetative identity. These results are discussed in relation to knowledge accumulated in Arabidopsis and to domestication processes.     

Apical Growth and Modification of the Development of Primordia During Re-flowering of Reverted Plants of Impatiens balsamina L

Impatiens balsamina L. was induced to flower by exposure to5 short days and then made to revert to vegetative growth byreturn to long days. After 9 long days reverted plants wereinduced to re-flower by returning them to short days. Petalinitiation began immediately and seven primordia already presentdeveloped into petals instead of into predominantly leaf-likeorgans. However, the arrangement of primordia at the shoot apex,their rate of initiation and size at initiation remained unchangedfrom the reverted apex, as did apical growth rate and the lengthof stem frusta at initiation. The more rapid flowering of thereverted plants than of plants when first induced, and the lackof change in apical growth pattern, imply that the revertedapices remain partially evoked, and that the apical growth patternand phyllotaxis typical of the flower, and already present inthe reverted plants, facilitate the transition to flower formation. Impatiens balsamina, flower reversion, partial evocation, shoot meristem, determination, leaf development     

Simulation of organ patterning on the floral meristem using a polar auxin transport model

An intriguing phenomenon in plant development is the timing and positioning of lateral organ initiation, which is a fundamental aspect of plant architecture. Although important progress has been made in elucidating the role of auxin transport in the vegetative shoot to explain the phyllotaxis of leaf formation in a spiral fashion, a model study of the role of auxin transport in whorled organ patterning in the expanding floral meristem is not available yet. We present an initial simulation approach to study the mechanisms that are expected to play an important role. Starting point is a confocal imaging study of Arabidopsis floral meristems at consecutive time points during flower development. These images reveal auxin accumulation patterns at the positions of the organs, which strongly suggests that the role of auxin in the floral meristem is similar to the role it plays in the shoot apical meristem. This is the basis for a simulation study of auxin transport through a growing floral meristem, which may answer the question whether auxin transport can in itself be responsible for the typical whorled floral pattern. We combined a cellular growth model for the meristem with a polar auxin transport model. The model predicts that sepals are initiated by auxin maxima arising early during meristem outgrowth. These form a pre-pattern relative to which a series of smaller auxin maxima are positioned, which partially overlap with the anlagen of petals, stamens, and carpels. We adjusted the model parameters corresponding to properties of floral mutants and found that the model predictions agree with the observed mutant patterns. The predicted timing of the primordia outgrowth and the timing and positioning of the sepal primordia show remarkable similarities with a developing flower in nature.     

Effect of decapitation, phosfon D and cycocel on the flowering of Impatiens balsamina exposed to varying numbers of short days

Impatiens balsamina L., a qualitative short day plant, requiresmore short days for the development of floral buds into flowersthan for their initiation. Phosfon D and cycocel reduce thenumber of short days required for flowering, increase the numberof floral buds and flowers and delay their reversion to vegetativegrowth when transferred to noninductive conditions. The effectof decapitation of the main shoot subsequent to the emergenceof floral buds resembles that of retardants indicating thatthe effect of the latter in flower promotion in this plant maybe by virtue of their effect on cessation of apical dominanceas a consequence of which reserve food materials may be channeledto axillary floral buds enabling them to develop into flowers. (Received January 9, 1969; )     

Two lily SEPALLATA-like genes cause different effects on floral formation and floral transition in Arabidopsis

Two AGL2-like MADS-box genes, Lily MADS Box Gene (LMADS) 3 and LMADS4, with extensive homology of LMADS3 to the Arabidopsis SEPALLATA3 were characterized from the lily (Lilium longiflorum). Both LMADS3 and LMADS4 mRNA were detected in the inflorescence meristem, in floral buds of different developmental stages, and in all four whorls of the flower organ. LMADS4 mRNA is also expressed in vegetative leaf and in the inflorescence stem where LMADS3 expression is absent. Transgenic Arabidopsis, which ectopically expresses LMADS3, showed novel phenotypes by significantly reducing plant size, flowering extremely early, and loss of floral determinacy. By contrast, 35S::LMADS4 transgenic plants were morphologically indistinguishable from wild-type plants. The early-flowering phenotype in 35S::LMADS3 transgenic Arabidopsis plants was correlated with the up-regulation of flowering time genes FT, SUPPRESSOR OF OVEREXPRESSION OF CO 1, LUMINIDEPENDENS, and flower meristem identity genes LEAFY and APETALA1. This result was further supported by the ability of 35S::LMADS3 to rescue the late-flowering phenotype in gigantea-1 (gi-1), constans-3 (co-3), and luminidependens-1 but not for ft-1 or fwa-1 mutants. The activation of these flowering time genes is, however, indirect because their expression was unaffected in plants transformed with LMADS3 fused with rat glucocorticoid receptor in the presence of both dexamethasone and cycloheximide.     

In Vitro Flowering Induction in Date Palm (Phoenix dactylifera L.)

This article reports on a histological and morphological study on the induction of in vitro flowering in vegetatively propagated plantlets from different date palm cultivars. The study aimed to further explore the control of in vitro flower induction in relation to the photoperiodic requirements in date palm and to come up with a novel system that may allow for early sex determination through plant cycle reduction. In fact, the in vitro reversion of a shoot meristem from a vegetative to a reproductive state was achieved within 1–5 months depending on the variety considered. This reversion was accompanied by several morphological transformations that affected the apical part of the leafy bud corresponding mainly to a size increase of the prefloral meristem zone followed by the appearance of an inflorescence. The flowers that were produced in vitro were histologically and morphologically similar to those formed in vivo. The histological examination of the in vitro flowering induction process showed that the conversion into inflorescences involved the entire apical vegetative meristem of the plantlet used as a starting material and brought about a change in its anatomical structure without affecting its phyllotaxis and the leaf shape. Through alternating between hormone-free and hormone-containing media under different light/dark conditions, the highest flower induction rates were obtained with a basal Murashige and Skoog medium. A change in the architectural model of date palm was induced because unlike the natural lateral flowering, in vitro flowering was terminal. Such in vitro flower induction allowed a significant reduction in plant cycle and can, therefore, be considered a promising candidate to save time for future improvement and selection programs in date palm.     

Auxin Depletion from the Leaf Axil Conditions Competence for Axillary Meristem Formation in Arabidopsis and Tomato

The enormous variation in architecture of flowering plants is based to a large extent on their ability to form new axes of growth throughout their life span. Secondary growth is initiated from groups of pluripotent cells, called meristems, which are established in the axils of leaves. Such meristems form lateral organs and develop into a side shoot or a flower, depending on the developmental status of the plant and environmental conditions. The phytohormone auxin is well known to play an important role in inhibiting the outgrowth of axillary buds, a phenomenon known as apical dominance. However, the role of auxin in the process of axillary meristem formation is largely unknown. In this study, we show in the model species Arabidopsis thaliana and tomato (Solanum lycopersicum) that auxin is depleted from leaf axils during vegetative development. Disruption of polar auxin transport compromises auxin depletion from the leaf axil and axillary meristem initiation. Ectopic auxin biosynthesis in leaf axils interferes with axillary meristem formation, whereas repression of auxin signaling in polar auxin transport mutants can largely rescue their branching defects. These results strongly suggest that depletion of auxin from leaf axils is a prerequisite for axillary meristem formation during vegetative development.     

Genetics of flower initiation and development in annual and perennial plants

Flowering is an integral developmental process in angiosperms, crucial to reproductive success and continuity of the species through time. Some angiosperms complete their life cycle within a year (annual plants), and others have a longer reproductive life, which is characterized by the generation of new flowering and vegetative shoots every year (perennial plants). Despite the differences in their lifespan, the underlying genetics of flower induction and floral organ formation appears to be similar among these plants. Hence, the knowledge gained from the study of flowering mechanism in Arabidopsis thaliana can be used to better understand similar processes in other plant species, especially the perennials, which usually have a long generation time and are not amenable to genetic analysis. Using Arabidopsis as a model, we briefly discuss the current understanding of the transition from vegetative to reproductive growth and the subsequent formation of individual floral organs, and how this knowledge has been successfully applied to the identification of homologous genes from perennial crops. Although annuals appear to share many similarities with perennials in terms of gene function, they differ in their commitment to flowering. Once an annual reaches the reproductive phase, all meristems are typically converted into either floral or inflorescence meristems. In contrast, each year, each meristem of a mature perennial has the choice to produce either a vegetative or a reproductive shoot. The physiology and genetics of flowering in Citrus are used to highlight the complexity of reproductive development in perennials, and to discus possible future research directions.