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Polyploidy is an important evolutionary phenomenon but the mechanisms by which polyploidy arises still remain underexplored. There may be an environmental component to polyploidization. This study aimed to clarify how temperature may promote diploid gamete formation considered an essential element for sexual polyploidization. First of all, a detailed cytological analysis of microsporogenesis and microgametogenesis was performed to target precisely the key developmental stages which are the most sensitive to temperature. Then, heat-induced modifications in sporad and pollen characteristics were analysed through an exposition of high temperature gradient. Rosa plants are sensitive to high temperatures with a developmental sensitivity window limited to meiosis. Moreover, the range of efficient temperatures is actually narrow. 36 °C at early meiosis led to a decrease in pollen viability, pollen ectexine defects but especially the appearance of numerous diploid pollen grains. They resulted from dyads or triads mainly formed following heat-induced spindle misorientations in telophase II. A high temperature environment has the potential to increase gamete ploidy level. The high frequencies of diplogametes obtained at some extreme temperatures support the hypothesis that polyploidization events could have occurred in adverse conditions and suggest polyploidization facilitating in a global change context.

This study presents the isolation and characterization of a novel nonsymbiotic Hb gene from sessile oak (Quercus petraea) seedlings, herein designated QpHb1. The cellular and tissue expression of QpHb1 was analysed by Northern blotting and in situ hybridization. The encoded protein was predicted to consist of 161 amino acid residues, and shares 71 and 51% amino acid sequence identity with the Arabidopsis class 1 and 2 nonsymbiotic Hb, respectively. Northern blot analysis revealed that QpHb1 was strongly expressed in roots. Spatial expression analysis of QpHb1 in the root apical region of sessile oak by in situ hybridization indicated that transcripts were mostly abundant in protoxylem cell initials, some cortical cells and the protoderm. In addition, when comparing the expression profile of QpHb1 in sessile and pedunculate oak (Quercus robur), two species with contrasted hypoxia tolerance, the transcript level of QpHb1 rose early in the most flood-tolerant species, pedunculate oak, during root submergence. The spatial-temporal expression of QpHb1 suggests that this gene could participate in perception and signalling during hypoxia.

Sessile oak is a species of great ecological and silvicultural importance in Europe; however, increased frequency and severity of flooding of forested areas pose a threat to its regeneration. We monitored water relations, root anatomical changes and the expression of two calmodulin genes (QpCaM) in sessile oak seedlings during a 14-day flooding treatment. The response followed two characteristic sequences. The first phase, in response to between 1 h and 3 days of flooding, was characterized by a dramatic but transient decline in water relations parameters followed by a recovery towards control values with no noticeable change in root cell morphology. During the second phase, in response to 3 to 14 days of flooding, water relations parameters gradually and continuously declined and hypertrophied lenticels developed at the base of the shoot. Concurrently, root cortical cells became larger and less spherical and the root cortex more porous. These cellular changes were accompanied by a transient rise in root transcript levels of QpCaM-2. We conclude that sessile oak seedlings are capable of withstanding a 3-day period of flooding without significant morphological alterations. In contrast, exposure to flooding for more than 3 days resulted in anatomical and morphological changes in the root system. These changes are, however, insufficient to provide sessile oak with long-term tolerance to flooding.

Astragalus tragacantha is a protected plant species in France that grows even in the trace metal and metalloid (TMM) polluted soils of the Calanques National Park (PNCal). Soils are mainly contaminated by lead, copper, zinc and arsenic. An ex situ experiment was conducted, firstly to determine the molecular responses and root traits involved in the TMM tolerance of this plant species by growing individuals in a soil from the surroundings of one of the brownfields of the PNCal, known as l'Escalette, where this plant species grows spontaneously. Secondly, in order to determine the plasticity of these responses, seeds were collected from three different populations, one at l'Escalette (polluted site), one from the Frioul archipelago (non-polluted, insular site) and one from La Seyne (non-polluted, littoral site). The results of this study confirmed the capacity of A. tragacantha to germinate and grow in TMM contaminated soils. Only moderate significant variations in chlorophyll and flavonol indices, proline content and antioxidant activities were detected between polluted and control soil conditions for all populations. The main driver for A. tragacantha TMM tolerance seemed to be its ability to be associated with root symbionts i.e. arbuscular mycorrhizal fungi and dark septate endophytes, corresponding to a nutrient-uptake strategy trait. This work provides support for the challenge of A. tragacantha conservation along the littoral of the PNCal, because increasing the number of A. tragacantha individuals would both increase vegetation cover of the polluted soils to reduce the pollution transfer and reinforce the populations of this species. Highlights A rare plant species able to grow in metal and metalloid contaminated soils Germination and growth not affected by Pb and As soil pollution High plasticity of seedlings of this rare protected plant Enhanced Pb and As tolerance due to nutrient-uptake strategy Preserving this rare species may improve phytostabilization of soil pollutants

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Recherche, for its assistance and logistical support in carrying out this 23 study. The authors would also like to thank Michael Paul for revising the English of this text 24 and are grateful to the two anonymous reviewers for their suggestions that helped to improve 25 the quality of the manuscript.

Recent studies consider the native flora as a potential source of plant candidates for phytostabilization of metal and metalloid (MM) contaminated soils, but ecological restoration is not the main objective of these researches. However, in contaminated areas, phytostabilization should be considered as a useful tool for ecological restoration. The present study takes stock of 3 years of a Mediterranean pilot site implementation using native plant species to recover plant and microbial communities (diversity and functions) together with soil remediation in the Calanques National Park. To determine the success of this operation, three in situ treatments were compared: ecological restoration plots characterized by the handling of physical environment (creation of cultivation terraces) and plants, negative control plots without vegetation but with the same physical environment handling as ecological restoration plots, and positive control plots with natural vegetation and no *Revised Manuscript Click here to view linked References 2 handling. The results suggest that an ecological restoration trajectory is initiated in the ecological restoration plots, characterized by a partial permanent plant cover. However, there is no evidence of a significant improvement of soil quality (evaluated by soil texture, pH, nutrients and organic carbon contents, cation exchange capacity, microbial biomass and activities) and phytostabilization efficiency after 3 years. Native plant communities and their associated microorganisms may need more time before improving soil quality and MM stabilization under the drastic Mediterranean conditions. Any amendment addition to accelerate restoration and MM immobilization was forbidden in this protected area. Under such conditions, an active restoration need to be carried out in this contaminated area even if resilient dynamics of the native plant communities may sporadically occur over a long period of time.