Transformation of chicory and expression of the bacterial uidA and nptll genes in the transgenic regenerants

Transgenic chicory plants were obtained from different explantsco-cultured with Agrobacterium tumefaciens. Among tap-root,leaf and cotyledonary tissues, etiolated cotyledons showed thegreatest competence for transformation. The Agrobacterium strainsused contained either pGSGLUC1 or pTDE4 as a vector which carryboth the neomycin phosphotransferase II gene (nptll) for kanamycinresistance and ß-glucuronidase gene (uidA) under thecontrol of different promoters. Transformation was confirmedby NPTII enzymatic assay, histochemical analysis of GUS activityand DNA hybridization. Transgenic plants expressed both markergenes in root and shoot tissues. In leaves, GUS activity wasexpressed in all tissue types, whatever the nature of the promoter.Nevertheless, variable heterogeneous patterns of expressionwere observed in the different root tissues. Differential expression of the GUS fusions controlled by thedual TR or the CaMV 35S promoters are discussed. Key words: Chicory, genetic transformation, GUS activity, kanamycin resistance     

Clonal Growth Strategies Along Flooding and Grazing Gradients in Atlantic Coastal Meadows

Specific composition and species clonal traits were characterized along combined flooding and grazing gradients to answer two questions. i) To what extent does the interaction of flooding and grazing influence the clonal characteristics of the vegetation? ii) Are the effects of both environmental factors independent or interactive? This study was carried out in a wet meadow along the Atlantic coast (France). Three plant communities (hygrophilous, meso-hygrophilous and mesophilous) were distinguished along a flooding gradient and five levels of grazing pressure were controlled through an experimental design (from no grazing to heavy grazing). We monitored species composition and retrieved, for each species, the type of clonal growth organs (CGOs) and clonal traits from the CLO-PLA3 database. We identified two syndromes of clonal traits: ??above-ground splitters?? and ??below-ground integrators??. Clonal traits played a key role in plant assembly in the studied meadows. The interaction of both environmental factors selected for particular syndromes of clonal traits; however, flooding had a stronger filtering effect than grazing. The hygrophilous community was dominated by above-ground splitters, whereas the meso-hygrophilous vegetation was dominated by below-ground integrators. In the mesophilous community, clonal composition was the most diverse and shared clonal traits with the vegetation of both the hygrophilous and meso-hygrophilous communities. Grazing impact on CGOs and clonal traits differed between plant communities, i.e., the effect of grazing was modulated by the flooding regime. This study confirmed that vegetation responses to grazing might depend on the pool of traits, primarily filtered by environmental factors such as flooding.     

Stronger Short-Term Effects of Mowing Than Extreme Summer Weather on a Subalpine Grassland

Mowing is known to favor plant diversity and influence ecosystem functioning in semi-natural grasslands. This effect could be influenced by climate variability, especially in regions with harsh climate, such as subalpine zones. In particular, short-term extreme weather fluctuations may induce rapid plant responses, affecting in turn the response to mowing. We tested the effects of concomitant summer weather manipulation and mowing on a subalpine grassland in the Central French Alps for two consecutive years. We addressed two questions: (1) How is a subalpine grassland affected by extreme summer weather? (2) Does extreme summer weather alter mowing effects on the grassland plant diversity and functioning? We used a multi-level, integrative approach assessing the responses of six abundant plant species, as well as effects on plant community structure, biomass production, and litter decomposition rates. Extreme summer weather was simulated by increasing summer temperature by 1.1°C, and decreasing summer rainfall by 80%—resulting in a 30% decrease in total annual precipitation. In addition, a heat-wave event was simulated during the first year of the experiment. This weather manipulation was combined with a late-summer mowing treatment (mown vs. unmown). Extreme summer weather mainly increased leaf senescence and decreased plant vegetative growth. Leaf litter decomposition was slowed, but only for species characterized by the fastest rates of litter decomposition. Mowing increased plant diversity by restricting the dominant grass species, thereby favoring subordinates. In the short term, this subalpine grassland was rather resistant to extreme summer weather, whereas mowing cessation remained the main factor affecting its biodiversity.     

Responses of clonal architecture to experimental defoliation: a comparative study between ten grassland species

Clonal architecture may enable plants to effectively respond to environmental constraints but its role in plant tolerance to defoliation remains poorly documented. In several non-clonal species, modifications of plant architecture have been reported as a mechanism of plant tolerance to defoliation, yet this has been little studied in clonal plants. In a glasshouse experiment, five rhizomatous and five stoloniferous species of grazed pastures were subjected to three frequencies of defoliation in order to test two hypotheses. (1) We expected plant clonal response to defoliation to be either a more compact architecture (low clonal propagation, but high branching), or a more dispersed one (long-distance propagation and low branching). Such plastic adjustments of clonal architecture were assumed to be involved in tolerance to defoliation i.e. to promote genet performance in terms of biomass and number of ramets. (2) The response of clonal architecture to defoliation was expected to be dependent on the species and to be more plastic in stoloniferous than in rhizomatous species. Most genets of each species were tolerant to defoliation as they survived and developed in every treatment. Architectural modifications in response to defoliation did not match our predictions. Clonal growth was either maintained or reduced under defoliation. Relative growth rate (RGR) decreased in eight species, whereas defoliated genets of seven species produced as many ramets as control genets. Biomass allocation to ramet shoots remained stable for all but one species. In defoliated genets, the number and mean length of connections, and mean inter-ramet distance were equal to or lower than those in control genets. Four groups of species were distinguished according to their architectural response to defoliation and did not depend on the type of connections. We hypothesised that dense clonal architectures with low plasticity may be the most advantageous response in defoliated conditions such as in grazed pastures. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Influence of abscisic acid on nitrogen partitioning, sucrose metabolism and nitrate reductase activity of chicory suspension cells

Batch suspension cultures of chicory cells (Cichorium intybusL. var. Witloof) possess a NADH-specific nitrate reductase activitythat peaks on day 3 of a 10 d growth cycle. When both nitrateand ammonium are used as nitrogen sources, chicory cells absorbnitrate irst. Ammonium uptake becomes predominant at day 3,even though NO₃^– was still present in the medium. Althoughabscisic acid impairs growth as well as ¹⁵NO₃^– uptakeand reduction, it promotes nitrate reductase activity as measuredboth in vivo and in vitro. Specific activity is 50% higher inABA-treated cells than in controls. These conflicting data maybe explained either in erms of nitrate reductase levels or bythe availability of reducing power and energy. Since NRA isgenerally controlled by the availability of the reducing power,the energy status of the cell, the adenylate nucleotide pools,were measured simultaneously with the carbohydrate levels withinthe cell and the growth medium. The energy charge was not modifiedduring the growth cycle, regardless of the rowth conditions.Yet ABA modified the intracellular carbohydrate metabolism andinhibited the acidic invertase, the sucrose synthase and thesucrose phosphate synthase activities. Modified assimilationrates of nitrate in chicory cells grown in the presence of ABA,were probably correlated to modified carbohydrate metabolismpathways leading to increased availability of reducing power,energy and C-skeletons. Key words: Abscisic acid, Cichorium intybus L, nitrate reductase, reductase, invertase, sucrose synthase, sucrose phosphate synthase     

Consequence of ramet defoliation on plant clonal propagation and biomass allocation: Example of five rhizomatous grassland species

We investigated the effects of ramet defoliation frequency on clonal propagation and the patterns of biomass production and allocation on five rhizomatous species (Carex divisa Hude., Eleocharis palustris L., Juncus articulatus L., Juncus gerardii Lois. and Elytrigia repens L.). Plants were grown during an 18-week experiment in greenhouse conditions. The above ground parts of ramets were clipped following three treatments: frequent (every 2 weeks), moderate (every 4 weeks) and unclipped (control). The growth of C. divisa, J. articulatus and E. repens was strongly affected by defoliation whereas E. palustris and J. gerardii maintained a similar performance when defoliated. The latter were able to compensate for the biomass loss even after six consecutive clippings. Defoliation frequency had a significant effect on total biomass production for C. divisa, J. articulatus and E. repens while J. gerardii and E. palustris maintained total biomass production. Most of the studied species showed a decrease in clonal traits when defoliated. Clipped plants displayed fewer and shorter rhizomes. Defoliation had a strong influence in biomass production with a decrease in rhizome mass in all clipped species. A greater allocation to aerial parts and a lower to rhizomes were also detected. Moderate defoliation entailed intermediate response in 1/3 of detected significant effects of defoliation on plant traits. Finally, in the experimental conditions, E. palustris and J. gerardii were the most tolerant species to defoliation, while J. articulatus was intermediate and C. divisa and E. repens had the lowest tolerance.     

The effects of management on vegetation trajectories during the early‐stage restoration of previously arable land after hay transfer

The restoration of floodplain grasslands has benefited from many studies of the underlying mechanisms. Among the operational tools that resulted, hay transfer is now used increasingly to alleviate the effects of limited seed dispersal and recruitment. To improve this method, we still need to understand how it can affect restoration trajectories, and particularly their direction and magnitude during the early stages of restoration. Based on concepts from the field of community ecology theory, we investigated the effects of early‐stage management through grazing or mowing on restoration trajectories after soil harrowing and hay transfer. We established a randomized block design experiment and quantified several community‐related metrics to formalize restoration trajectories for 3 years after hay transfer on a previously arable alluvial island in southwestern France. Whatever the management treatment, the species richness and evenness were significantly higher in hay‐inoculated than in control plots. This effect was linked to the recruitment of species originating not only from the reference grassland through hay transfer, but also from the seed bank, a well‐known effect of soil harrowing. Although generally oriented toward the reference grassland, the origin, direction, and magnitude of the trajectory of hay‐inoculated plots all depended on the management applied. Sheep grazing applied at the same time as hay transfer enhanced the recruitment of reference species as from the first experimental year, because it controlled aboveground competition and maintained the window of opportunity open for a sufficiently longer period of time. Our findings show that the type of management applied simultaneously to hay transfer influences the origin of a grassland trajectory, while its direction and magnitude are dependent on the management applied in subsequent years. Grazing immediately after hay transfer may be appropriate to accelerate the recruitment of species from the reference grassland.     

Do spatial patterns of clonal fragments and architectural responses to defoliation depend on the structural blue-print? An experimental test with two rhizomatous Cyperaceae

Clonal architecture is involved in performance of clonal fragments, as it determines spatial distribution of ramets. It is expected to rely on the species-specific expression of several architectural traits (structural blue-print). However, in contrasting environments, realized clonal architectures may differ, due to phenotypic plasticity. In this paper, we compared clonal architectures between two rhizomatous ecologically close Cyperaceae (Carex divisa and Eleocharis palustris) in non-defoliated and defoliated conditions. Two questions were addressed. (1) How much do the structural blue-print and resulting colonization and occupation of space differ between both species? (2) Does the structural blue-print constrain plastic responses of clonal architecture to defoliation? Traits related to performance, spatial pattern, architecture and biomass allocation of clonal fragments were monitored through an original non-destructive mapping method. In non-defoliated conditions, both species showed similar biomass but contrasting architectures and patterns of biomass allocation to rhizomes that resulted in different spatial patterns. The rhizome network of C. divisa, which consisted in only two primary rhizomes but several branches, was involved in resource storage rather than in spatial colonization. Conversely, E. palustris produced on average six primary rhizomes that grew in the whole horizontal plane, maximizing both occupation and colonization of space. These differences in structural constraints coupled with allometric relationships, resulted in differential responses to defoliation. In C. divisa, the costs associated to defoliation caused a decrease in branching, limiting the area occupied and number of ramets produced by clonal fragments, but increasing ramet density. Conversely, the weakly branched rhizome network of E. palustris was not affected by defoliation. Both spatial strategies (consolidation vs. colonization) are likely to provide ecological advantages allowing their coexistence in grazed meadows.