Floral Stimulus Research Articles

Regulation of tissue repair in plants

A recent report in PNAS by Asahina et al. (1) addresses the fascinating question of tissue repair in plants. According to recent suggestions, plants and animals might share cellular mechanisms that allow regeneration of tissues after damage (2). However, plants and animals differ greatly in their mode of development and their ability to respond to damage-inducing environmental factors (3). Terrestrial plants cannot move their whole body in response to environmental cues, and, because of their cell walls, they also lack cellular mobility within the plant. This means that plants must regenerate damaged tissue through cellular regeneration at the point of damage. Traditionally, this regeneration was considered to occur by dedifferentiation of existing mature cells followed by cell division to form callus and differentiation to form the cellular constituents of the new tissue, although details of this process have been questioned recently (4). Plants experience many types of tissue damage, including that caused by herbivory and other forms of physical wounding (e.g., breakage because of wind or ice or trampling by animals). They have developed elaborate responses to this damage. For example, herbivory results in a suite of responses; some are fast-acting and local, whereas others may be quite long-lived and systemic in nature, allowing the plant to develop a response at the whole-plant level to attack by particular animal species (5). One of the simplest forms of damage to plants is the splitting or laceration of tissue. This type of wounding is frequent under both natural and agronomic conditions. It is also common with some of our well-established horticultural and research techniques (e.g., grafting). Indeed, grafting and the subsequent tissue repair have been vital for the identification of two new plant hormones over the last 5 years: the strigolactones for branching (6) and the floral stimulus or florigen … [↵][1]1To whom correspondence should be addressed. E-mail: Jim.Reid{at}utas.edu.au. [1]: #xref-corresp-1-1

Juvenility and flowering of Brunonia australis (Goodeniaceae) and Calandrinia sp. (Portulacaceae) in relation to vernalization and daylength

The time at which plants are transferred to floral inductive conditions affects the onset of flowering and plant morphology, due to juvenility. Plants of Brunonia australis and Calandrinia sp. were used to investigate whether Australian native ephemeral species show a distinct juvenile phase that can be extended to increase vegetative growth and flowering. The juvenile phase was quantified by transferring seedlings from less inductive (short day and 30/20°C) to inductive (vernalization or long day) conditions at six different plant ages ranging from 4 to 35 d after seed germination. An increase in days to first visible floral bud and leaf number were used to signify the end of juvenility. Brunonia australis was receptive to floral inductive long day conditions about 18-22 d after seed germination, whereas plants aged 4-35 d appeared vernalization sensitive. Overall, transferring plants of B. australis from short to long day conditions reduced the time to anthesis compared with vernalization or constant short day conditions. Calandrinia sp. showed a facultative requirement for vernalization and an insensitive phase was not detected. Floral bud and branch production increased favourably as plant age at time of transfer to inductive conditions increased. Younger plants showed the shortest crop production time. Both species can perceive the vernalization floral stimulus from a very young age, whereas the photoperiodic stimulus is perceived by B. australis after a period of vegetative growth. However, extending the juvenile phase can promote foliage development and enhance flower production of both species.

Effect of Different Leaf Removal Treatments on Flowering in Certain Early Flowering Sugarcane Varieties

Field experiments were conducted to determine the effects on flowering responses of the removal of selected leaves in sugarcane varieties during the induction period. It was found that the spindle leaf had a predominant role in the production of floral stimulus. The removal of leaf spindle once during the induction period, i.e. first week of August resulted in reduced flowering intensity and delayed flowering. The removal of all the leaves except the spindle leaf resulted in earlier flowering as compared to the undefoliated control. The removal of last exposed dewlap leaf (LED) and LED + 1, however, did not influence or affect flowering.

A cis Element within Flowering Locus T mRNA Determines Its Mobility and Facilitates Trafficking of Heterologous Viral RNA

The Arabidopsis flowering locus T (FT) gene encodes the mobile florigen essential for floral induction. While movement of the FT protein has been shown to occur within plants, systemic spread of FT mRNA remains to be unequivocally demonstrated. Utilizing novel RNA mobility assay vectors based on two distinct movement-defective viruses, Potato virus X and Turnip crinkle virus, and an agroinfiltration assay, we demonstrate that nontranslatable FT mRNA, independent of the FT protein, moves throughout Nicotiana benthamiana and mutant Arabidopsis plants and promotes systemic trafficking of viral and green fluorescence protein RNAs. Viral ectopic expression of FT induced flowering in the short-day N. tabacum Maryland Mammoth tobacco under long-day conditions. Recombinant Potato virus X bearing FT RNA spread and established systemic infection more quickly than the parental virus. The cis-acting element essential for RNA movement was mapped to the nucleotides 1 to 102 of the FT mRNA coding sequence. These data demonstrate that a plant self-mobile RNA molecule can mediate long-distance trafficking of heterologous RNAs and raise the possibility that FT RNA, along with the FT protein, may be involved in the spread of the floral stimulus throughout the plant.

Immediate induction of APETALA1-like gene expression following a single short-day in Pharbitis nil

Pharbitis [Ipomoea] nil cv. Violet is an excellent model plant for the study of photoperiodic induction of flowering because it can be induced flowering by a single short-day. However there are few molecular-level studies of the induction of flowering in Pharbitis. To gain insight into the photoperiodic induction of flowering, we isolated an APETALA1-like gene (PnAP1), which showed high similarity to SQUAMOSA (SQUA) and AP1. PnAP1 expression at the shoot apex was induced only under flowering-inductive conditions, and PnAP1 expression was induced from 24 h after the start of the inductive dark period. Both night-break and low ambient temperature during the dark period effectively repressed PnAP1 expression and flowering. Timing of PnAP1 expression assessed through induced cotyledon removal indicated that the floral stimulus began to move from cotyledons 14 h after the start of the inductive dark period. The results indicate that floral transitions begin immediately after the inductive dark period and that PnAP1 is a good molecular marker of floral transition.

FT Protein Acts as a Long-Range Signal in Arabidopsis

Plants are sessile organisms and must respond to changes in environmental conditions. Flowering time is a key developmental switch that is affected by both day length and temperature. Environmental cues are sensed by the leaves while the responses occur at the apex, requiring long-range communication within the plant. For many years it has been known that leaves exposed to light can trigger the floral transition of a darkened shoot, and grafting experiments demonstrated that the floral stimulus travels long distances. This mobile signal was later termed "florigen," but its nature has been unclear. The gene FLOWERING LOCUS T (FT) is a major output of both the photoperiod and the vernalization pathways controlling the floral transition. FT protein acts at the shoot apex of the plant in concert with a transcription factor, FLOWERING LOCUS D (FD). A fundamental question is how FT transcription in the leaves leads to active FT protein at the apex. We have uncoupled FT protein movement from its biological function to show that FT protein is the mobile signal that travels from the leaves to the apex. To our knowledge, FT is the only known protein that serves as a long-range developmental signal in plants.

The control of flowering in time and space

The transition to flowering is one of the most important developmental decisions made by plants. Classical studies have highlighted the importance of photoperiod in controlling flowering time. More recently, the identification of mutants specifically affected in the photoperiod pathway in the model system Arabidopsis thaliana has enabled the flowering time pathways to be placed in a molecular context. This review highlights recent advances in understanding how photoperiod signals (perceived in the leaves) act at the apex of the plant where the floral stimulus is perceived. The photoperiod pathway acts predominantly through the gene CONSTANS to activate the small signalling molecule FT. While FT transcription is induced in the leaves, it is essential that FT protein is present at the apex of the plant. FT at the apex interacts with the transcription factor FD to induce flowering.

The quest for florigen: a review of recent progress

The photoperiodic induction of flowering is a systemic process requiring translocation of a floral stimulus from the leaves to the shoot apical meristem. In response to this stimulus, the apical meristem stops producing leaves to initiate floral development; this switch in morphogenesis involves a change in the identity of the primordia initiated and in phyllotaxis. The physiological study of the floral transition has led to the identification of several putative floral signals such as sucrose, cytokinins, gibberellins, and reduced N-compounds that are translocated in the phloem sap from leaves to the shoot apical meristem. On the other hand, the genetic approach developed more recently in Arabidopsis thaliana allowed the discovery of many genes that control flowering time. These genes function in 'cascades' within four promotive pathways, the 'photoperiodic', 'autonomous', 'vernalization', and 'gibberellin' pathways, which all converge on the 'integrator' genes SUPPRESSOR OF OVEREXPRESSION OF CO 1 (SOC1) and FLOWERING LOCUS T (FT). Recently, several studies have highlighted a role for a product of FT as a component of the floral stimulus or 'florigen'. These recent advances and the proposed mode of action of FT are discussed here.

Florigen goes molecular: Seventy years of the hormonal theory of flowering regulation

As early as in 1936, the comprehensive studies of flowering led M.Kh. Chailakhyan to the concept of florigen, a hormonal floral stimulus, and let him establish several characteristics of this stimulus. These studies set up for many years the main avenues for research into the processes that control plant flowering, and the notion of florigen became universally accepted by scientists worldwide. The present-day evidence of genetic control of plant flowering supports the idea that florigen participates in floral signal transduction. The recent study of arabidopsis plants led the authors to conclusion that the immediate products of the gene FLOWERING LOCUS I, its mRNA and/or protein, move from an induced leaf into the shoot apex and evoke flowering therein.

The Message Is the Messenger

As the days lengthen in spring and summer, plants sense the hours of daylight in their leaves and respond by initiating flowering at the top of the plant, in the shoot apex. The identity of the signal that is transported from the leaf to the apex has been unclear (see Wigge et al. , Abe et al. , and the Perspective by Blázquez in the 12 August issue). Huang et al. now show that local expression of the gene FT ( FLOWERING LOCUS T ) in a single leaf is sufficient to cause flowering by transfecting a heat-inducible form of FT into Arabidopsis plants. Within 6 hours of stimulating a single leaf, FT messenger RNA (mRNA) appears in the shoot apex, where it stimulates transcription of genes involved in flowering and of FT itself. Although other elements may be involved, FT mRNA is an important component of the floral stimulus that moves from leaf to shoot in response to increases in day length. P. A. Wigge, M. C. Kim, K. E. Jaeger, W. Busch, M. Schmid, J. U. Lohmann, D. Weigel, Integration of spatial and temporal information during floral induction in Arabidopsis . Science 309 , 1056-1059 (2005). [Abstract] [Full Text] M. Abe, Y. Kobayashi, S. Yamamoto, Y. Daimon, A. Yamaguchi, Y. Ikeda, H. Ichinoki, M. Notaguchi, K. Goto, T. Araki, FD, a bZIP protein mediating signals from the floral pathway integrator FT at the shoot apex. Science 309 , 1052-1056 (2005). [Abstract] [Full Text] M. A. Blázquez, The right time and place for making flowers. Science 309 , 1024-1025 (2005). [Summary] [Full Text] T. Huang, H. Böhlenius, S. Eriksson, F. Parcy, O. Nilsson, The mRNA of the Arabidopsis gene FT moves from leaf to shoot apex and induces flowering. Science 309 , 1694-1696 (2005). [Abstract] [Full Text]

Photoperiodic flowering of Arabidopsis: integrating genetic and physiological approaches to characterization of the floral stimulus

ABSTRACTIn many plants the transition from vegetative growth to flowering is controlled by environmental cues. One of these cues is day length or photoperiod, which synchronizes flowering of many species with the changing seasons. Recently, advances have been made in understanding the molecular mechanisms that confer photoperiodic control of flowering and, in particular, how inductive events occurring in the leaf, where photoperiod is perceived, are linked to floral evocation that takes place at the shoot apical meristem. We discuss recent data obtained using molecular genetic approaches on the function of regulatory proteins that control flowering time in Arabidopsis thaliana. These data are compared with the results of physiological analyses of the floral transition, which were performed in a range of species and directed towards identification of the transmitted floral singals.

Isolation and Characterization of LEAFY and APETALA1 Homologues from Citrus sinensis L. Osbeck `Washington'

Homologues of the floral meristem identity genes LEAFY (LFY) and APETALA1 (AP1) were isolated from the hybrid perennial tree crop `Washington' navel orange (Citrus sinensis) and designated CsLFY and CsAP1, respectively. Citrus has an extended juvenile period unlike herbaceous plants and responds to different floral stimuli than herbaceous plants or deciduous tree species. Despite these differences, the deduced amino acid sequences of CsLFY and CsAP1 genes are at least 65% identical with their Arabidopsis thaliana L. Heynh counterparts and share even greater sequence similarity to LFY and AP1 from the deciduous woody perennials, Populus balsamifera Bradshaw and Populus tremuloides Michaux, respectively. Like A. thaliana LFY (AtLFY) and AP1 (AtAP1), CsLFY and CsAP1 expression was restricted almost exclusively to reproductive tissues, but observed expression of CsAP1 in the fourth whorl carpel tissue of mature flowers was distinct from other plant AP1 genes. Transgenic A. thaliana plants ectopically expressing CsLFY or CsAP1 showed early-flowering phenotypes similar to those described for overexpression of AtLFY and AtAP1. In addition, the 35S:CsLFY and 35S:CsAP1 transgenes partially complemented the lfy-10 and ap1-3 mutants, respectively. The severity of the overexpression phenotypes correlated with the accumulation of CsLFY or CsAP1 transcripts. LFY is a single-copy gene in flowering plants but consistent with its hybrid origin, the genome of C. sinensis `Washington' has two easily distinguishable CsLFY and CsAP1 alleles derived from it's parental genotypes, C. maxima L. Osbeck (pummelo) and C. reticulata Blanco (mandarin). Allelic polymorphism at both the CsLFY and CsAP1 loci was restricted to the 5′- and 3′-flanking regions.

CONSTANS acts in the phloem to regulate a systemic signal that induces photoperiodic flowering of Arabidopsis.

Flower development at the shoot apex is initiated in response to environmental cues. Day length is one of the most important of these and is perceived in the leaves. A systemic signal, called the floral stimulus or florigen, is then transmitted from the leaves through the phloem and induces floral development at the shoot apex. Genetic analysis in Arabidopsis identified a pathway of genes required for the initiation of flowering in response to day length. The nuclear zinc-finger protein CONSTANS (CO) plays a central role in this pathway, and in response to long days activates the transcription of FT, which encodes a RAF-kinase-inhibitor-like protein. We show using grafting approaches that CO acts non-cell autonomously to trigger flowering. Although CO is expressed widely, its misexpression from phloem-specific promoters, but not from meristem-specific promoters, is sufficient to induce early flowering and complement the co mutation. The mechanism by which CO triggers flowering from the phloem involves the cell-autonomous activation of FT expression. Genetic approaches indicate that CO activates flowering through both FT-dependent and FT-independent processes, whereas FT acts both in the phloem and the meristem to trigger flowering. We propose that, partly through the activation of FT, CO regulates the synthesis or transport of a systemic flowering signal, thereby positioning this signal within the established hierarchy of regulatory proteins that controls flowering.

Cell division and morphological changes in the shoot apex of Arabidopsis thaliana during floral transition.

Eight-week-old vegetative plants of Arabidopsis thaliana, ecotype Columbia, were induced to flower by a single long day (LD). In this experimental system, it is known that the last component of the floral stimulus moves from the leaves to the apex 24-36 h after the start of the LD, and the first floral meristem is initiated by the shoot apical meristem (SAM) at 44-56 h (Corbesier et al., 1996, The Plant Journal 9: 947-952). Here we show that the rate of cell division is increased at floral transition in all SAM parts but not in the sub-apical pith cells. Mitotic activity starts to increase 24 h after the start of the LD and is two- to three-fold higher at peak times than that in non-induced plants. This activation is followed by the start of SAM enlargement at 44 h, SAM doming at 48 h, and the elongation of apical internodes (bolting) at 52 h.

Synthesis of gibberellin GA 6 and its role in flowering of Lolium temulentum

The induction of flowering by one long day (LD) in the grass Lolium temulentum is most closely mimicked by application of the gibberellins (GAs) GA 5 or GA 6, both of which occur naturally. These gibberellins promote floral development but have little effect on stem elongation. Endogenous GA 5 and GA 6 contents in the shoot apex double on the day after the LD and, for GA 5 (and we presume for GA 6 as well) reach a concentration known to be inductive for the excised shoot apex in vitro. They are, therefore, strong candidates as LD floral stimuli in this grass. The synthesis of GA 6 and an examination of its florigenic properties in L. temulentum are described.

Long-Day Induction of Flowering in Lolium temulentumInvolves Sequential Increases in Specific Gibberellins at the Shoot Apex

One challenge for plant biology has been to identify floral stimuli at the shoot apex. Using sensitive and specific gas chromatography-mass spectrometry techniques, we have followed changes in gibberellins (GAs) at the shoot apex during long day (LD)-regulated induction of flowering in the grass Lolium temulentum. Two separate roles of GAs in flowering are indicated. First, within 8 h of an inductive LD, i.e. at the time of floral evocation, the GA(5) content of the shoot apex doubled to about 120 ng g(-1) dry weight. The concentration of applied GA(5) required for floral induction of excised apices (R.W. King, C. Blundell, L.T. Evans [1993] Aust J Plant Physiol 20: 337-348) was similar to that in the shoot apex. Leaf-applied [(2)H(4)] GA(5) was transported intact from the leaf to the shoot apex, flowering being proportional to the amount of GA(5) imported. Thus, GA(5) could be part of the LD stimulus for floral evocation of L. temulentum or, alternatively, its increase at the shoot apex could follow import of a primary floral stimulus. Later, during inflorescence differentiation and especially after exposure to additional LD, a second GA action was apparent. The content of GA(1) and GA(4) in the apex increased greatly, whereas GA(5) decreased by up to 75%. GA(4) applied during inflorescence differentiation strongly promoted flowering and stem elongation, whereas it was ineffective for earlier floral evocation although it caused stem growth at all times of application. Thus, we conclude that GA(1) and GA(4) are secondary, late-acting LD stimuli for inflorescence differentiation in L. temulentum.

Hormone physiology of pea mutants prevented from flowering by mutations gi or veg1.

The veg1 (vegetative) mutant in pea (Pisum sativum L.) does not flower under any circumstances and gi (gigas) mutants remain vegetative under certain conditions. gi plants are deficient in production of floral stimulus, whereas veg1 plants lack a response to floral stimulus. During long days in particular, these non-flowering mutant plants eventually enter a stable compact phase characterised by a large reduction in internode length, small leaves and growth of lateral shoots from the upper-stem (aerial) nodes. The first-order laterals in turn produce second-order laterals and so on in a reiterative pattern. The apical bud is reduced in size but continues active growth. Endogenous hormone measurements and gibberellin application studies with gi-1, gi-2 and veg1 plants indicate that a reduction in gibberellin and perhaps indole-3-acetic acid level may account, at least partially, for the compact aerial shoot phenotype. In the gi-1 mutant, the compact phenotype is rescued by transfer from a 24- to an 8-h photoperiod. We propose that in plants where flowering is prevented by a lack of floral stimulus or an inability to respond, the large reduction in photoperiod gene activity during long days may lead to a reduction in apical sink strength that is manifest in an altered hormone profile and weak apical dominance.

Long-Day Induction of Flowering in Lolium temulentum Involves Sequential Increases in Specific Gibberellins at the Shoot Apex

One challenge for plant biology has been to identify floral stimuli at the shoot apex. Using sensitive and specific gas chromatography-mass spectrometry techniques, we have followed changes in gibberellins (GAs) at the shoot apex during long day (LD)-regulated induction of flowering in the grass Lolium temulentum. Two separate roles of GAs in flowering are indicated. First, within 8 h of an inductive LD, i.e. at the time of floral evocation, the GA5 content of the shoot apex doubled to about 120 ng g−1 dry weight. The concentration of applied GA5 required for floral induction of excised apices (R.W. King, C. Blundell, L.T. Evans [1993] Aust J Plant Physiol 20: 337–348) was similar to that in the shoot apex. Leaf-applied [2H4] GA5 was transported intact from the leaf to the shoot apex, flowering being proportional to the amount of GA5 imported. Thus, GA5 could be part of the LD stimulus for floral evocation of L. temulentum or, alternatively, its increase at the shoot apex could follow import of a primary floral stimulus. Later, during inflorescence differentiation and especially after exposure to additional LD, a second GA action was apparent. The content of GA1 and GA4 in the apex increased greatly, whereas GA5 decreased by up to 75%. GA4 applied during inflorescence differentiation strongly promoted flowering and stem elongation, whereas it was ineffective for earlier floral evocation although it caused stem growth at all times of application. Thus, we conclude that GA1 and GA4are secondary, late-acting LD stimuli for inflorescence differentiation in L. temulentum.

N content of phloem and xylem exudates during the transition to flowering in Sinapis alba and Arabidopsis thaliana

ABSTRACTThe involvement of nitrogenous substances in the transition to flowering was investigated in Sinapis alba and Arabidopsis thaliana (Columbia). Both species grown in short days (SD) are induced to flower by one long day (LD). In S. alba, the phloem sap (leaf and apical exudates) and the xylem sap (root exudate) were analysed in LD versus SD. In A. thaliana, only the leaf exudate could be analysed but an alternative system for inducing flowering without day‐length extension was used: the displaced SD (DSD). Significant results are: (i) in both species, the leaf exudate was enriched in Gln during the inductive LD, at a time compatible with export of the floral stimulus; (ii) in S. alba, the root export of amino acids decreased in LD, whereas the nitrate remained unchanged – thus the extra‐Gln found in the leaf exudate should originate from the leaves; (iii) extra‐Gln was also found very early in the apical exudate of S. alba in LD, together with more Glu; (iv) in A. thaliana induced by one DSD, the leaf export of Asn increased sharply, instead of Gln in LD. This agrees with Asn prevalence in C‐limited plants. The putative role of amino acids in the transition to flowering is discussed.

The frequency of plasmodesmata increases early in the whole shoot apical meristem of Sinapis alba L. during floral transition.

The frequency of plasmodesmata increases in the shoot apical meristem of plants of Sinapis alba L. induced to flower by exposure to a single long day. This increase is observed within all cell layers (L1, L2, L3) as well as at the interfaces between these layers, and it occurs in both the central and peripheral zones of the shoot apical meristem. The extra plasmodesmata are formed only transiently, from 28 to 48 h after the start of the long day, and acropetally since they are detectable in L3 4 h before they are seen in L1 and L2. These observations indicate that (i) in the Sinapis shoot apical meristem at floral transition, there is an unfolding of a single field with increased plasmodesmatal connectivity, and (ii) this event is an early effect of the arrival at this meristem of the floral stimulus of leaf origin. Since (i) the wave of increased frequency of plasmodesmata is 12 h later than the wave of increased mitotic frequency (A. Jacqmard et al. 1998, Plant cell proliferation and its regulation in growth and development, pp. 67 78; Wiley), and (ii) the increase in frequency of plasmodesmata is observed in all cell walls, including in walls not deriving from recent divisions (periclinal walls separating the cell layers), it is concluded that the extra plasmodesmata seen at floral transition do not arise in the forming cell plate during mitosis and are thus of secondary origin.