Repression of floral meristem fate is crucial in shaping tomato inflorescence.

Tomato is an important crop and hence there is a great interest in understanding the genetic basis of its flowering. Several genes have been identified by mutations and we constructed a set of novel double mutants to understand how these genes interact to shape the inflorescence. It was previously suggested that the branching of the tomato inflorescence depends on the gradual transition from inflorescence meristem (IM) to flower meristem (FM): the extension of this time window allows IM to branch, as seen in the compound inflorescence (s) and falsiflora (fa) mutants that are impaired in FM maturation. We report here that Jointless (J), which encodes a MADS-box protein of the same clade than Short Vegetative Phase (SVP) and Agamous Like 24 (AGL24) in Arabidopsis, interferes with this timing and delays FM maturation, therefore promoting IM fate. This was inferred from the fact that j mutation suppresses the high branching inflorescence phenotype of s and fa mutants and was further supported by the expression pattern of J, which is expressed more strongly in IM than in FM. Most interestingly, FA--the orthologue of the Arabidopsis LEAFY (LFY) gene--shows the complementary expression pattern and is more active in FM than in IM. Loss of J function causes premature termination of flower formation in the inflorescence and its reversion to a vegetative program. This phenotype is enhanced in the absence of systemic florigenic protein, encoded by the Single Flower Truss (SFT) gene, the tomato orthologue of Flowering Locus T (FT). These results suggest that the formation of an inflorescence in tomato requires the interaction of J and a target of SFT in the meristem, for repressing FA activity and FM fate in the IM.

Flowering response of the uniflora:blind:self-pruning and jointless:uniflora:self-pruning tomato (Solanum lycopersicum) triple mutants.

Tomato (Solanum lycopersicum) is a day-neutral plant with a sympodial growth habit. Triple mutants were produced in order to investigate the genetic interactions in the flowering regulation of this species in the initial and sympodial segments. The jointless:uniflora:self-pruning and uniflora:blind:self-pruning triple mutants were produced by crossing the jointless:uniflora and uniflora:blind double mutants with the uniflora:self-pruning double mutant. The phenotype of the triple mutants was characterized and the expression of the affected genes was studied in the uniflora (uf) mutant through semi-quantitative reverse transcriptase polymerase chain reaction (PCR). The triple mutants produced solitary flowers, as their uf parent, instead of inflorescences. They were both late flowering in the initial segment with a flowering time intermediate between their parental double mutants. The flowering time of the sympodial segments was delayed in the jointless:uniflora:self-pruning triple mutant while the uniflora:blind:self-pruning triple mutant did not initiate sympodial segments. The expression of the studied genes was not markedly affected by the uf mutation. These results suggest that floral transition of the primary shoot and of sympodial segments is regulated differently in tomato. The UNIFLORA (UF) gene acts upstream of the other investigated genes in controlling reproductive structure and flowering time of the initial segment although their expression does not seem to be affected by the uf mutation. In the sympodial segments, the self-pruning determinate phenotype is strengthened by the blind mutation and suppressed by the jointless mutation.

Impact of salinity on early reproductive physiology of tomato (Solanum lycopersicum) in relation to a heterogeneous distribution of toxic ions in flower organs.

The effect of short-term treatments (10 days) by a high salt level (150 mm NaCl) on vegetative and reproductive development was investigated in tomato plants (Solanum lycopersicum L. cv. Ailsa Craig) at two developmental stages. Salinity applied during flowering transition reduced shoot biomass and delayed the appearance of the first inflorescence. Both shoot and root biomasses were reduced when salt was applied during the development of the first inflorescence. At both stages, areas of young leaves decreased and time to first anthesis increased, while total number of flowers in the first inflorescence was not affected. Flower abortion, reduction of pollen number and viability were higher when salinity was applied during inflorescence development. Na+ accumulated in all organs while K+ decreased. Laser ablation inductively coupled plasma mass spectrometry microanalysis revealed that Na+ accumulated in style, ovaries and anther intermediate layers but not in the tapetum nor in the pollen grains when salinity was applied during inflorescence development. K+ was not significantly affected in these structures. Soluble carbohydrates dramatically increased in leaves and decreased in the inflorescence under salt stress conditions. The failure of inflorescence to develop normally under salt stress can be better explained in terms of altered source-sink relationships rather than accumulation of toxic ions.

Morphology of nectaries and biology of nectar production in the distylous species Fagopyrum esculentum.

BACKGROUND AND AIMS: The mechanisms of floral nectar production in buckwheat (Fagopyrum esculentum, Polygonaceae), a distylous pseudo-cereal, have received relatively little attention, prompting an investigation of the factors that regulate this process. The aim was to perform a refined study of the structures that secrete nectar and of the internal and external parameters influencing nectar volumes and sugar concentrations. METHODS: In order to control environmental parameters, plants were cultivated in growth rooms under controlled conditions. The structure of nectaries was studied based on histological sections from flowers and flower buds. Nectar was extracted using glass micropipettes and the sugar concentration was measured with a hand refractometer. Sugar concentration in the phloem sap was measured using the anthrone method. To test the influence of photosynthesis on nectar production, different light and defoliation treatments were applied. KEY RESULTS: Unicellular trichomes were located in the epidermis at the ventral part of eight nectary glands situated on the flower receptacle alternately with stamens. Vascular bundles consisting of both phloem and xylem were identified at the boundary between a multilayered nectary parenchyma and a sub-nectary parenchyma with chloroplasts. A higher volume of nectar in thrum morphs was observed. No other difference was found in morphology or in sugar supply to inflorescences between morphs. Nectar secretion was strongly influenced by plant age and inflorescence position. Nectar volumes were higher in the upper inflorescences and during the flowering peak. Light had a dual role, (1) acting directly on reproductive structures to trigger flower opening, which conditions nectar secretion, and (2) stimulating photosynthetic activity, which regulates nectar accumulation in open flowers. CONCLUSIONS: In buckwheat, nectar is secreted by trichomes and probably proceeds, at least in part, from phloem sap. Nectar secretion is strongly influenced by floral morph type, plant age, inflorescence position and light.

Resource availability regulates reproductive meristem activity, development of reproductive structures and seed set in buckwheat (Fagopyrum esculentum).

Grain yields in buckwheat (Fagopyrum esculentum) are consistently low despite a profuse and long-lasting flowering. The mechanisms underlying this reproductive failure are still largely unknown. In this study, performed in strictly controlled conditions, the possible implication of assimilate availability in the regulation of the reproductive development was investigated in the 'La Harpe' cultivar, by manipulating sink/source relationships through excisions of selected leaves and/or selected inflorescences and hand pollinations of selected and precisely identified flowers. Increasing or decreasing source/sink organ ratio in buckwheat, respectively, enhanced or reduced the production of racemes by the shoot apical meristem and of cymes by the racemes, ultimately modulating the number of flowers per plant. The effect on seed production was essentially indirect and related to flower production. Seeds developing on a plant also affected reproductive morphogenesis, limiting flower production. Three stages at which flower development to mature seed may fail have been identified. First, an abnormal morphogenesis resulted in flowers with an undersized and sterile gynoecium. Secondly, after anthesis, numerous flowers with a normal sized gynoecium, were unable to develop a seed after hand pollination. Finally, a small proportion of flowers exhibited a later abortion of the seed. Flower fate is dependent on position in the raceme and time of opening: chance to develop normally and produce a ripe seed is higher for flowers at the base than at the top of the inflorescence. A mechanism, internal to the raceme, regulates flower fate, independently of assimilate availability. This mechanism may be disturbed by drastic reductions in nutrient supply.

Genetic interactions in the control of flowering time and reproductive structure development in tomato (Solanum lycopersicum).

Different tomato (Solanum lycopersicum) mutants, affected in flowering time, reproductive structure or plant architecture, were crossed to produce double mutants in order to investigate gene interactions in flowering regulation in this autonomous species with a sympodial growth habit. The compound inflorescence: uniflora, uniflora: self pruning, uniflora: blind, and jointless: uniflora double mutants all produced solitary flowers like their uniflora parent, instead of inflorescences. All double mutants were late flowering. uniflora: blind and uniflora: self pruning had flowering times intermediate between those of their two parents. jointless: uniflora and compound inflorescence: uniflora flowered later than uniflora, the mutant with the most delayed flowering. All double mutants developed strong lateral shoots at node levels approximately corresponding to the level at which their parent cultivars initiated their first reproductive structure, which is a typical trait of uniflora. These results suggest that the UNIFLORA gene acts upstream of the other investigated genes in controlling flowering in tomato, and that floral transition of the primary shoot and floral transition of sympodial segments are regulated differently.

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.

NaCl alleviates polyethylene glycol-induced water stress in the halophyte species Atriplex halimus L.

Atriplex halimus L. is a C4 xero-halophyte species well adapted to salt and drought conditions. To collect information on the physiological impact of low salt levels on their water-stress resistance, seedlings were exposed for 6 d to nutrient solution containing either 0% or 15% polyethylene glycol 10,000 (PEG), in the presence or in the absence of 50 mM NaCl. Similar experiments were performed with one PEG-resistant and one PEG-sensitive selected cell line exposed for 50 d to 0% or 15% PEG on standard Linsmaier and Skoog (LS) medium, on LS medium supplemented with 50 mM NaCl, or on Na+-free medium. NaCl mitigated the deleterious impact of PEG on growth of both whole plants and PEG-sensitive cell lines and improved the ability of stressed tissues to perform osmotic adjustment (OA). Water stress reduced CO2 net assimilation rates quantified in the presence of high CO2 and low O2 levels (A), stomatal conductance and transpiration, but NaCl improved water use efficiency of PEG-treated plants through its positive effect on A values, especially in young leaves. PEG increased the internal Na+ concentration. The resistant cell line accumulated higher concentration of Na+ than the PEG-sensitive one. The complete absence of Na+ in the medium endangered the survival of both cell lines exposed to PEG. Although Na+ by itself contributed only for a small part to OA, NaCl induced an increase in proline concentration and stimulated the synthesis of glycinebetaine in response to PEG in photosynthetic tissues. Soluble sugars were the main contributors to OA and increased when tissues were simultaneously exposed to PEG and NaCl compared with PEG alone, suggesting that Na+ may influence sugar synthesis and/or translocation.

Is osmotic adjustment required for water stress resistance in the Mediterranean shrub Atriplex halimus L?

The effect of water stress was investigated in plants from two populations of Atriplex halimus L: Tensift issued from a salt-affected coastal area and Kairouan, originating from an inland dried site. Water deficit was applied by withholding water for 22 days. Shoot dry weight (shoot DW), leaf relative water content (RWC), turgid weight to dry weight ratio (TW/DW), osmotic potential (psis), osmotic adjustment (OA), proline, glycinebetaine, and sugar content were determined 1, 8, 15 and 22 days after withholding watering. Water stress induced a decrease in shoot DW, RWC, psis, and TW/DW, but an increase in glycinebetaine and sugar leaf contents. The decrease of psis and TW/DW was more marked in Kairouan than in Tensift. At the end of the stress period, Kairouan showed a greater OA compared with Tensift. However, the contribution of net solute accumulation (OAacc) was similar in both populations in response to stress. Water stress resistance could thus not be associated with higher OA, although the ability of plants to regulate these metabolic and physiological functions could play an important role under harmful conditions. The possible roles of osmolyte accumulations are discussed in relation to the specific physiological strategy of water-stress-resistance in this species.

Inflorescence structure and control of flowering time and duration by light in buckwheat (Fagopyrum esculentum Moench).

Morphogenesis of the reproductive structure of buckwheat and the impact of light conditions on flowering time and duration have been investigated using the variety 'La Harpe'. Inflorescences were initiated acropetally, in leaf axils, by the shoot apical meristem until its arrest of functioning which was accompanied by the abortion of the last inflorescence produced. The buckwheat inflorescence is a compound raceme that produces laterally flowered cymose clusters, the number of which was affected by the position of the inflorescence along the main stem. Similarly, the number of flowers in a lateral cluster was dependent on the inflorescence's position on the stem. The development of each inflorescence was stopped as its meristem stopped functioning and, in a situation reminiscent of the shoot apical meristem, the latest initiated cyme aborted. The development of each cyme was also terminated with the abortion of a few young flowers. The variety 'La Harpe' is a facultative short-day plant: the number of nodes generated before the initiation of the first inflorescence and the number of days from sowing to macroscopic appearance of this inflorescence were reduced in 8 h days as compared with 16 h days. The number of inflorescences, and thus flowering duration, was also strongly reduced by short days. It was unaffected by light irradiance in 8 h days while, in 16 h days, it was prolonged when light intensity was increased, suggesting the interaction of two different mechanisms for its regulation. Buckwheat is a distylous species, but inflorescence structure and flowering behaviour were not affected by floral morph.

UNIFLORA, a pivotal gene that regulates floral transition and meristem identity in tomato (Lycopersicon esculentum).

• Flowering of uniflora (uf), a tomato (Lycopersicon esculentum) mutant which consistently produces solitary flowers instead of inflorescences, is late and highly asynchronous in winter. This puzzling behaviour prompted us to further investigate flowering regulation in this mutant to improve our understanding of UNIFLORA gene function. • Growing plants under different daylengths and light intensities revealed that flowering time in uf is dependent on daily light energy integral. Transferring plants from low to high light energy integrals at different times after sowing showed that the light-conditions effect was stage dependent, suggesting that interactions between light energy integrals and endogenous regulatory pathways affect meristem sensitivity to flowering signals. • Carbohydrate analyses suggested that one of these signals could be sucrose, but other interacting factors are probably generated by the root system, as indicated by grafting experiments. • The UNIFLORA gene thus appears to have a dual role in tomato: floral transition regulation and the maintenance of inflorescence meristem identity.

Flower architecture and sex determination: how does Atriplex halimus play with floral morphogenesis and sex genes?

• Atriplex halimus , a monoecious Chenopodiaceae , produces flowers displaying two basic architectures. • The first architectural pattern is made of staminate pentamerous flowers with an external whorl of yellowish tepals and an internal whorl of stamens. The second architectural pattern consists of female flowers with a single carpel enclosed within two opposite bracts. In both architectures, bisexual flowers and flowers of the un-expected sex were detected leading to the occurrence of up to six floral phenotypes on the same individual. • Daylength and light intensity affected sex ratio and flower distribution between both architectural patterns. Short days and low light irradiance promoted femaleness and bracteate floral architecture. Flower position on a reproductive axis and geographical origin of the plant (genotype) also affected sex and architecture ratios. • Thus, all the genetic information required for the production of both floral architecture and sexual organ types is present in each A. halimus plant but endogenous and environmental cues determine the fate of the floral meristems. These results are discussed in relation to classical models of genetic control of floral morphogenesis.