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.     

Strigolactones promote nodulation in pea

Strigolactones are recently defined plant hormones with roles in mycorrhizal symbiosis and shoot and root architecture. Their potential role in controlling nodulation, the related symbiosis between legumes and Rhizobium bacteria, was explored using the strigolactone-deficient rms1 mutant in pea (Pisum sativum L.). This work indicates that endogenous strigolactones are positive regulators of nodulation in pea, required for optimal nodule number but not for nodule formation per se. rms1 mutant root exudates and root tissue are almost completely deficient in strigolactones, and rms1 mutant plants have approximately 40% fewer nodules than wild-type plants. Treatment with the synthetic strigolactone GR24 elevated nodule number in wild-type pea plants and also elevated nodule number in rms1 mutant plants to a level similar to that seen in untreated wild-type plants. Grafting studies revealed that nodule number and strigolactone levels in root tissue of rms1 roots were unaffected by grafting to wild-type scions indicating that strigolactones in the root, but not shoot-derived factors, regulate nodule number and provide the first direct evidence that the shoot does not make a major contribution to root strigolactone levels.     

Flowering pathway is regulated by bulb size in Lilium longiflorum (Easter lily)

Lilium longiflorum (Easter lily) vegetative propagation occurs through production of underground bulbs containing apical and axillary meristems. In addition, sexual reproduction is achieved by flowering of elongated shoots above the bulb. It is generally accepted that L. longiflorum has an obligatory requirement for vernalisation and that long day (LD) regime hastens flowering. However, the effect of bulb size and origin, with respect to axillary or apical meristems on flowering, as well as the interactions between these meristems are largely unknown. The aim of this study was to explore the effect of bulb size, vernalisation and photoperiod on L. longiflorum flowering. To this end, we applied vernalisation and photoperiod treatments to the different bulb sizes and used a system of constant ambient temperature of 25 °C, above vernalisation spectrum, to avoid cold‐dependent floral induction during plant growth. Vernalisation and LD hasten flowering in all bulbs. Large, non‐vernalised bulbs invariably remained at a vegetative stage. However, small non‐vernalised bulbs flowered under LD conditions. These results demonstrate for the first time that cold exposure is not an obligatory prerequisite for L. longiflorum flowering, and that an alternative flowering pathway can bypass vernalisation in small bulbs. We suggest that apical dominance interactions determine the distinct flowering pathways of the apical and axillary meristems. Similar floral induction is achieved in propagated bulblets from scaling. These innovative findings in the field of geophyte floral induction represent valuable applicative knowledge for lily production.     

Effects of some growth regulators on in vitro flowering of Streptocarpus nobilis

Leaf discs from vegetative Streptocarpus nobilis plants were cultured in vitro in media with cytokinin (BAP or K at 0.35 mg.1^?1) and auxin (IAA, NAA or 2,4-D at 0.1 mg.1^?1). Under short days (8-h photoperiod) in medium with IAA and BAP, floral buds developed in 100% of the cultures; under long days (16-h photoperiod) only shoots were formed. In medium with IAA and K, flowering was reduced. Flowers rarely formed in medium containing NAA and K, but roots developed profusely. NAA + BAP promoted leafy shoots which rarely flowered later. The effect of 2,4-D was to inhibit flowering completely and to induce callusing and formation of teratomous structures.     

Auslösung der Blütenbildung bei der Langtagpflanze Hyoscyamus niger im Kurztag durch 2-Thiouracil

Summary Aqueous solutions of 2-thiouracil (TU) were applied selectively either to the growing point or to the leaves of the long-day plant Hyoscyamus niger in order to determine whether this antimetabolite has an effect on the synthesis of the floral stimulus in the leaves. Applications to the growing point were made by means of a small glass tube covering the shoot apex; application to the leaves was performed by vacuum infiltration. In all experiments all leaves except the three youngest fully expanded leaves and the 8–10 youngest primordia were removed before application. Plants were recorded as having initiated flowers when flower primordia were visible under a dissection microscope 5 weeks after the experiment.TU was inhibitory to photoperiodic induction by long-days of 16 hours when applied to the growing point during the second 8 hours of the daily photoperiod. A concentration of 5·10^-3 M of TU fully suppressed flowering without significant inhibition of leaf primordia increment; however, leaves developing from treated primordia had reduced leaf blades. These results are in agreement with findings already published by other investigators.However, when the leaves were infiltrated by TU, the antimetabolite did not inhibit photoperiodic induction but on the contrary initiated flowering even under short-day conditions. This effect was investigated in more detail by repeated daily infiltrations of TU-solutions in concentrations of 10^-5–10^-2 M during the second part of an 8 hour photoperiod up to 5 following days. Even after one single infiltration of a 10^-4 M solution 18% of the treated plants were flowering; the percentage of flowering plants increased with increasing concentrations of TU and number of days of application up to approximately 80%. In no case was a flower initiation of 100% obtained. Leaves developing from primordia after infiltration of the leaves with TU have reduced and deformed leaf blades, indicating that TU is transported to the shoot apex to some extent.Some possible explanations of this inductive effect of TU were tested experimentally. Oxygen uptake of the leaves was not decreased and the respiratory quotient was not affected by TU. Photoperiodic induction is not stimulated by low concentrations of TU when applied to the growing point. Infiltration of the leaves by solutions of 2,4-dinitrophenol (10^-4 M) and sodium azide (10^-3 M) had no inductive effect under short-day conditions; a single complete defoliation (except for the 8–10 youngest primordia) is also not inductive. Under short-day conditions additional leaves remaining on the plant that were not infiltrated by TU decreased the percentage of flowering plants but did not fully suppress flower initiation.From these results it is concluded that TU does not act by inhibition of particular metabolic processes concerned in flower initiation or by inhibition of the synthesis of an inhibitor. We suggest that application of TU may lead to synthesis of a floral stimulus in the leaves under short-day conditions also.     

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.     

Role of VIN3-LIKE 2 in facultative photoperiodic flowering response in Arabidopsis

In Arabidopsis, expression of FLC and FLC-related genes (collectively called FLC clade) contributes to flowering time in response to environmental changes, such as day length and temperature, by acting as floral repressors. VIN3 is required for vernalization-mediated FLC repression and a VIN3 related protein, VIN3-LIKE 1/VERNALIZATION 5 (VIL1/VRN5), acts to regulate FLC and FLM in response to vernalization.^1–3 VIN3 also exists as a small family of PHD finger proteins in Arabidopsis, including VIL1/VRN5, VIL2/VEL1, VIL3/VEL2 and VIL4/VEL3. We showed that the PHD finger protein, VIL2, is required for proper repression of MAF5, an FLC clade member, to accelerate flowering under non-inductive photoperiods. VIL2 acts together with POLYCOMB REPRESSIVE COMPLEX 2 (PRC2) to repress MAF5 in a photoperiod dependent manner.Key words: photoperiod, chromatin, floweringThe decision to flower is critical to the survival of flowering plants. Thus, plants sense environmental cues to initiate floral transition at a time that both ensures and optimizes their own reproductive fitness. Using a model plant, Arabidopsis thaliana, genetic studies have shown that the regulation of floral transition mainly consists of four genetic pathways: the inductive photoperiod pathway, the autonomous pathway, the vernalization pathway and the gibberellin pathway.⁴ In Arabidopsis, these four flowering pathways eventually merge into a group of genes called floral integrators, including FLOWERING LOCUS T (FT), SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) and LEAFY (LFY). Based on the response to specific photoperiod conditions, the flowering behaviors of plants can be classified into three groups: long day (LD), short day (SD) and day neutral response.^5,6 Depending on the requirement of day length, plants show either obligate or facultative responses. For example, henbane, carnation and ryegrass are obligate long day (LD) flowering plants which flower under increasing inductive photoperiod but do not flower at all under non-inductive photoperiod.⁵ On the other hand, plants including Arabidopsis, wheat, lettuce and barley, are considered to be facultative flowering plants. Thus, these plants exhibit early flowering under LD and late-flowering under non-inductive short days (SD). Studies on photoperiodic flowering time mainly focus on the inductive LD-photoperiod pathway in Arabidopsis.     

Effect of Gibberellic Acid and Phosfon on the Critical Dark Period Reqnirement for Flowering of Impatiens balsamina cv. Rose

The critical dark period requirement for flowering of Impatiens balsamina L. cv. Rose, an obligate short day plant, is about 8.5 hours. While GA₃ completely substituted for the dark period requirement, Phosfon prolonged it to 9.5 hours. GA₃ hastened and Phosfon delayed the initiation of floral buds under all photoperiods. Floral buds opened into flowers only during 8 and 14 hour photoperiods in control and Phosfon-treated plants but during all photoperiods in GA₃-treated ones. The delay in floral bud initiation and flowering was correlated with shifting up of the node bearing the first floral bud and flower respectively. While GA₃ increased the numher of floral buds and flowers in all photoperiods except 8-hour, Phosfon increased their number in the 14-hour photoperiod only. The number of flowering plants decreased with increasing photoperiod regardless of GA₃ and Phosfon application. The effect of Phosfon was completely or partially overcome, depending upon the photoperiod, by simultaneous application of GA₃.     

Role of phytochrome B in organ formation processes in Cucumis sativus L.

The role of phytochrome B in the organogenesis process in the apical and axillary shoot meristems during early ontogenesis stages in cucumber Cucumis sativus L. at photoperiods (day/night) 10/14, 16/8 h, and continuous light in comparison with wild type plants and phytochrome B-deficient mutant (lh-mutant) was investigated. In mutant phytochrome B, deficiency caused inhibition of initiation of leaves both in the main shoot and lateral shoots and increased the number of flower buds (IV stage of organogenesis). With continuous light, the number of lateral shoots and flowers during stage IV of organogenesis in wild-type plants increased twofold in comparison with the mutant. Short-term temperature drops did not induce floral ontogenesis in mutants but increased the number of off-shoots in both experimental variants during a long photoperiod and continuous lighting. We propose that phytochrome B, by increasing the compactness of chromatin, may facilitate coordination of ontogenesis processes with changing environmental conditions.     

Bi-directional inflorescence development inArabidopsis thaliana: Acropetal initiation of flowers and basipetal initiation of paraclades

In this study we investigated Arabidopsis thaliana (L.) Heynh. inflorescence development by characterizing morphological changes at the shoot apex during the transition to flowering. Sixteen-hour photoperiods were used to synchronously induce flowering in vegetative plants grown for 30 d in non-inductive 8-h photoperiods. During the first inductive cycle, the shoot apical meristem ceased producing leaf primordia and began to produce flower primordia. The differentiation of paraclades (axillary flowering shoots), however, did not occur until after the initiation of multiple flower primordia from the shoot apical meristem. Paraclades were produced by the basipetal activation of buds from the axils of leaf primordia which had been initiated prior to photoperiodic induction. Concurrent with the activation of paraclades was the partial suppression of paraclade-associated leaf primordia, which became bract leaves. The suppression of bract-leaf primordia and the abrupt initiation of flower primordia during the first inductive photoperiod is indicative of a single phase change during the transition to flowering in photoperiodically induced Arabidopsis. Morphogenetic changes characteristic of the transition to flowering in plants grown continuously in 16-h photoperiods were qualitatively equivalent to the changes observed in plants which were photoperiodically induced after 30 d. These results suggest that Arabidopsis has only two phases of development, a vegetative phase and a reproductive phase; and that the production of flower primordia, the differentiation of paraclades from the axils of pre-existing leaf primordia and the elongation of internodes all occur during the reproductive phase.     

Preliminary characterization of floral response of <Emphasis Type="Italic">Xerophyta humilis</Emphasis> to desiccation,vernalisation, photoperiod and light intensity

Xerophyta humilis is a monocotyledonous resurrection plant found in arid and semi-arid summer rainfall areas of Southern Africa, which undergoes desiccation to survive periods of extreme drought. In order for X. humilis to thrive in their natural habitat, correct timing of the floral transition, coincident with wet periods of sufficient duration, is essential. In this study, the environmental cues involved in the regulation of the floral transition in X. humilis were analysed. No single parameter tested was sufficient to induce flowering, but it was found that flowering was promoted by a combination of a cool period experienced while plants were hydrated, followed by transfer to long-day photoperiods of relatively high light intensity. Plants retained competence to flower if desiccated during exposure to cold, but no flowering occurred if dried prior to this exposure. These data suggest that exposure to cold temperature facilitates vernalisation and subsequent exposure to high light and long days are inductive for floral initiation in X. humilis.     

Auslösung der Blütenbildung durch Cycloheximid im Kurztag bei der Langtagpflanze Hyoscyamus niger

Summary Cycloheximide (CH) was applied selectively either to the shoot apex or by infiltration to the leaves of the long-day plant Hyoscyamus niger in order to investigate whether this inhibitor has an effect on the synthesis of a floral stimulus in the leaves. Treatment of the shoot apex with CH caused inhibition of the photoperiodic induction. In contrast, when CH was applied to leaves, initiation of flowering was observed under short-day conditions. The drug yielded optimum initiating effects at concentrations of 10^-5-3·10^-5 M, inducing flowering of almost 60% of the plants. Daily infiltration over a period of up to 4 days decreased the rate of flower initiation. The effect of CH was shown to be additive to a photoperiodic induction, even to a sub-threshold induction, but not to 2-thiouracil mediated induction. In no case did the presence of additional untreated leaves on the plants suppress CH-mediated flower induction. Treatment of the leaves with chloramphenicol (10^-6-2^-10^-4 M) or puromycin (5·10^-6-2·10^-4 M) caused no initiating response. The results are interpreted to mean that the presence of CH in the leaves may lead to the synthesis of a floral stimulus also under short-day conditions. This finding is similar to that reported previously in the case of the inductive effect of 2-thiouracil.

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Folgende Abkürzungen wurden verwendet 2-TU 2-Thiouracil - CH Cycloheximid - LT Langtag - DL Dauerlicht Herrn Prof. Dr. L. Brauner in Verehrung und Dankbarkeit zum 75. Geburtstag gewidmet.     

The pea GIGAS gene is a FLOWERING LOCUS T homolog necessary for graft-transmissible specification of flowering but not for responsiveness to photoperiod

Garden pea (Pisum sativum) was prominent in early studies investigating the genetic control of flowering and the role of mobile flowering signals. In view of recent evidence that genes in the FLOWERING LOCUS T (FT) family play an important role in generating mobile flowering signals, we isolated the FT gene family in pea and examined the regulation and function of its members. Comparison with Medicago truncatula and soybean (Glycine max) provides evidence of three ancient subclades (FTa, FTb, and FTc) likely to be common to most crop and model legumes. Pea FT genes show distinctly different expression patterns with respect to developmental timing, tissue specificity, and response to photoperiod and differ in their activity in transgenic Arabidopsis thaliana, suggesting they may have different functions. We show that the pea FTa1 gene corresponds to the GIGAS locus, which is essential for flowering under long-day conditions and promotes flowering under short-day conditions but is not required for photoperiod responsiveness. Grafting, expression, and double mutant analyses show that GIGAS/FTa1 regulates a mobile flowering stimulus but also provide clear evidence for a second mobile flowering stimulus that is correlated with expression of FTb2 in leaf tissue. These results suggest that induction of flowering by photoperiod in pea results from interactions among several members of a diversified FT family.     

Circadian rhythms and the induction of flowering in the long-day grass Lolium temulentum L.

Plants of Lolium temulentum L. strain Ceres were grown in 8-h short day (SD) for 45 d before being exposed either to a single long day (LD) or to a single 8-h SD given during an extended dark period. For LD induction, the critical photoperiod was between 12 and 14 h, and more than 16 h were needed for a maximal flowering response. During exposure to a single 24-h LD, the translocation of the floral stimulus began between the fourteenth and the sixteenth hours after the start of the light period, and was completed by the twenty-fourth hour. Full flowering was also induced by one 8-h SD beginning 4 or 28 h after the start of a 40-h dark period, i.e. by shifting 12 h forward or beyond the usual SD. The effectiveness of a so-called ‘displaced short day’ (DSD) was not affected by light quality and light intensity. With a mixture of incandescent and fluorescent lights at a total photosynthetic photon flux density of 400 μmol m⁻² s⁻¹, a 4-h light exposure beginning 4 h after the start of a 40-h dark period was sufficient to induce 100% flowering. The flower-inducing effect of a single 8-h DSD was also assessed during a 64-h dark period. Results revealed two maxima at a 20-h interval. This fluctuation in light sensitivity suggests that a circadian rhythm is involved in the control of flowering of L. temulentum.     

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

In 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.     

The role of EMF1 in regulating the vegetative and reproductive transition in Arabidopsis thaliana (Brassicaceae)

To understand the genetic regulation of vegetative to reproductive transition in higher plants, further characterization of the Arabidopsis mutant embryonic flower1, emf1, was conducted. Using three flowering symptoms, we showed that emf1 mutants could only grow reproductive and not rosette shoots under five different growth conditions. The mutant embryos did not produce the typical tunica–corpus shoot apical structures at the heart-, torpedo-, and mature stages. The divergent shoot apical development during mutant and wild-type embryogenesis indicated that the wild-type EMF1 gene was expressed in early embryogenesis. Mutations in the EMF1 gene affected the embryonic shoot apical development and caused the germinating embryo and regenerating callus to grow inflorescence, instead of rosette, shoots. Our results support the hypothesis that the EMF1 gene regulates the switch between vegetative and reproductive growth in Arabidopsis.     

Gene <Emphasis Type="Italic">TAENIATA</Emphasis> Is a Novel Negative Regulator of the <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> Homeobox Genes <Emphasis Type="Italic">KNAT1, KNAT2, KNAT6</Emphasis>, and <Emphasis Type="Italic">STM</Emphasis>

Mutant Arabidopsis thaliana taeniata (tae) plants are characterized by an altered morphology of leaves and the inflorescence. At the beginning of flowering, the inflorescence produces fertile flowers morphologically intermediate between a shoot and a flower. The recessive mutation tae also causes the formation of ectopic meristems and shoot rosettes on leaves. The expressivity of the mutant characters depend on the temperature and photoperiod. Analysis of the activity of KNOX class I genes in the leaves of the tae mutant has demonstrated the expression of genes KNAT2 and STM and an increase in the expression of genes KNAT1 and KNAT6 compared to wild-type leaves. These data indicate that the TAE gene negatively regulates the KNAT1, KNAT2, KNAT6, and STM genes.__________Translated from Genetika, Vol. 41, No. 8, 2005, pp. 1068–1074.Original Russian Text Copyright © 2005 by Lebedeva, Ezhova, Melzer.