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The capacity of marine sponges to cope with environmental changes is often attributed to the specific composition of their bacterial communities. In this study, we assessed the bacterial microbiome of two dominant sponges, Rhabdastrella globostellata (Rg) and Hyrtios erectus (He), living in the Bouraké lagoon (New Caledonia), where abiotic conditions daily fluctuate according to the tide. Sponge specimens, sediment and seawater samples were collected at 2–3 m depth. The bacterial communities were assessed using 16S rRNA metabarcoding, and variations between the two sponges were compared using PCA biplots. Chloroflexi was the dominant phyla in both He and Rg with an average relative abundance of 41.2% and 53.2%, respectively, while it was absent in sediment and seawater. Among the phylum Chloroflexi, SAR202 clade was dominant in both sponges, reaching an average relative abundance of 53.2% (He) and 78.7% (Rg). Principal component analysis (PCA) was used to identify the main variables driving the bacterial community structure in both sponges. The results indicated that the bacterial community structure in both sponges was strongly associated with Chloroflexi (70.9% of the phyla variance) and SAR202 clade (86.6% of the clade variance). The high relative abundance of the phylum Chloroflexi and the SAR202 clade observed in this study is the highest reported so far in the literature in marine sponges. Such a high relative abundance of these bacteria could suggest their involvement in the well-being of sponges in the extreme environmental conditions of Bouraké.

The ecological restoration of nickel mining-degraded areas in New Caledonia is strongly limited by low availability of soil mineral nutrients, metal toxicity, and slow growth rates of native plant species. In order to improve plant growth for restoration programs, special attention was paid to interactions between plant and soil microorganisms. In this study, we evaluated the influence of inoculation with Curtobacterium citreum BE isolated from a New Caledonian ultramafic soil on arbuscular mycorrhizal symbiosis and growth of Tetraria comosa, an endemic sedge used in restoration programs. A greenhouse experiment on ultramafic substrate was conducted with an inoculum comprising two arbuscular mycorrhizal fungi (AMF) species isolated from New Caledonian ultramafic soils: Rhizophagus neocaledonicus and Claroideoglomus etunicatum. The effects on plant growth of the AMF and C. citreum BE inoculated separately were not significant, but their co-inoculation significantly enhanced the dry weight of T. comosa compared with the non-inoculated control. These differences were positively correlated with mycorrhizal colonization which was improved by C. citreum BE. Compared with the control, co-inoculated plants were characterized by better mineral nutrition, a higher Ca/Mg ratio, and lower metal translocation. However, for Ca/Mg ratio and metal translocation, there were no significant differences between the effects of AMF inoculation and co-inoculation.

In New Caledonia, one third of the main island is covered by ultramafic soils characterized by high levels of metals and low levels of plant nutrients. In these soils, metal tolerant bacteria may play a role in plant adaptation to the edaphic constraints. Recently, two new bacteria species belonging to the genus Burkholderia sensu lato were isolated from rhizospheric roots of pioneer plant growing in New Caledonian ultramafic soils and were able to tolerate high metal concentrations. The purpose of this study was to determine the major mechanisms by which these two bacteria were able to tolerate metals and if they were able to reduce metal absorption by plants from which they were isolated. Bioaccumulation, adsorption on cell surface, ability to form biofilm and production of exopolysaccharides were assessed. A focus was carried out on Ni, generally the most toxic metal in New Caledonian ultramafic soils. All experiments were performed in comparison with a reference strain isolated from a non-ultramafic habitat, Paraburkholderia caribensis MWAP64T. The two ultramafic bacterial species showed a high tolerance to Ni with a Ni–IC50 of 5 mM for Caballeronia novacaledonica STM10272T and 30 mM for Paraburkholderia ultramafica STM10279T, values significantly higher than those of P. caribensis MWAP64T (2 mM). The major mechanism involved in the bacterial Ni tolerance was related to biofilm formation for C. novacaledonica STM10272T and to acidic exopolysaccharide production for P. ultramafica STM10279T. Both species alleviated metal contents in roots of Tetraria comosa, an endemic pioneer herbaceous species growing on New Caledonian ultramafic soils.

The present study focused on the characterization of 10

Acacia spirorbis subsp. spirorbis Labill. is a widespread tree legume endemic to New Caledonia that grows in ultramafic (UF) and volcano-sedimentary (VS) soils. The aim of this study was to assess the symbiotic promiscuity of A. spirorbis with nodulating and nitrogen-fixing rhizobia in harsh edaphic conditions. Forty bacterial strains were isolated from root nodules and characterized through (i) multilocus sequence analyses, (ii) symbiotic efficiency and (iii) tolerance to metals. Notably, 32.5% of the rhizobia belonged to the Paraburkholderia genus and were only found in UF soils. The remaining 67.5%, isolated from both UF and VS soils, belonged to the Bradyrhizobium genus. Strains of the Paraburkholderia genus showed significantly higher nitrogen-fixing capacities than those of Bradyrhizobium genus. Strains of the two genera isolated from UF soils showed high metal tolerance and the respective genes occurred in 50% of strains. This is the first report of both alpha- and beta-rhizobia strains associated to an Acacia species adapted to UF and VS soils. Our findings suggest that A. spirorbis is an adaptive plant that establishes symbioses with whatever rhizobia is present in the soil, thus enabling the colonization of contrasted ecosystems.

This study aims to characterize the ectomycorrhizal (ECM) communities associated with Acacia spirorbis, a legume tree widely spread in New Caledonia that spontaneously grows on contrasted edaphic constraints, i.e. calcareous, ferralitic and volcano-sedimentary soils. Soil geochemical parameters and diversity of ECM communities were assessed in 12 sites representative of the three mains categories of soils. The ectomycorrhizal status of Acacia spirorbis was confirmed in all studied soils, with a fungal community dominated at 92% by Basidiomycota, mostly represented by/tomentella-thelephora (27.6%), /boletus (15.8%), /sebacina (10.5%), /russula-lactarius (10.5%) and /pisolithus-scleroderma (7.9%) lineages. The diversity and the proportion of the ECM lineages were similar for the ferralitic and volcano-sedimentary soils but significantly different for the calcareous soils. These differences in the distribution of the ECM communities were statistically correlated with pH, Ca, P and Al in the calcareous soils and with Co in the ferralitic soils. Altogether, these data suggest a high capacity of A. spirorbis to form ECM symbioses with a large spectrum of fungi regardless the soil categories with contrasted edaphic parameters.

New Caledonian serpentine (ultramafic) soils contain high levels of toxic heavy metals, in particular nickel, (up to 20 g kg−1) and are deficient in essential elements like carbon, nitrogen and phosphorus while having a high magnesium/calcium ratio. Although previous studies showed that ectomycorrhizal symbioses could play an important role in the adaptation of the endemic plants to ultramafic soils (FEMS Microbiol Ecol 72:238–49, 2010), none of them have compared the diversity of microbial communities from ultramafic vs non-ultramafic soils in New Caledonia. We explored the impact of edaphic characteristics on the diversity of ectomycorrhizal (ECM) fungi associated with different endemic species of Tristaniopsis (Myrtaceae) growing under contrasting soil conditions in the natural ecosystems of New Caledonia. ECM root tips were thus sampled from two different ultramafic sites (Koniambo massif and Desmazures forest) vs two volcano-sedimentary ones (Arama and Mont Ninndo). The molecular characterization of the ECM fungi through partial sequencing of the ITS rRNA gene revealed the presence of different dominant fungal genera including, both soil types combined, Cortinarius (36.1%), Pisolithus (18.5%), Russula (13.4%), Heliotales (8.2%) and Boletellus (7.2%). A high diversity of ECM taxa associated with Tristaniopsis species was found in both ultramafic and volcano-sedimentary soils but no significant differences in ECM genera distribution were observed between both soil types. No link could be established between the phylogenetic clustering of ECM taxa and their soil type origin, thus suggesting a possible functional—rather than taxonomical—adaptation of ECM fungal communities to ultramafic soils.

The taxonomic status of eleven rhizospheric bacterial strains belonging to the genus Burkholderia and isolated from roots of Costularia (Cyperaceae), tropical herbaceous pioneer plants growing on ultramafic soils in New Caledonia, was investigated using a polyphasic taxonomic approach. The genetic analyses (16S rRNA genes, gyrB, recA, nreB and cnr) confirmed that all strains are Burkholderia and cluster into two separated groups. The DNA hybridization results showed low relatedness values to the closest relatives Burkholderia species. The phenotypic analyses confirmed that the two groups of strains could be differentiated from each other and from other known Burkholderia species. This polyphasic study revealed that these two groups of strains represent each a novel species of Burkholderia, for which the names Burkholderia novacaledonica sp. nov. (type strain STM10272(T)=LMG28615(T)=CIP110887(T)) and B. ultramafica sp. nov. (type strain STM10279(T)=LMG28614(T)=CIP110886(T)) are proposed, respectively. These strains of Burkholderia presented specific ecological traits such as the tolerance to the extreme edaphic constraints of ultramafic soils: they grew at pH between 4 and 8 and tolerate the strong unbalanced Ca/Mg ratio (1/19) and the high concentrations of heavy metals i.e. Co, Cr, Mn and Ni. Noteworthy B. ultramafica tolerated nickel until 10mM and B. novacaledonica up to 5mM. The presence of the nickel (nreB) and cobalt/nickel (cnr) resistance determinants encoding for protein involved in metal tolerance was found in all strains of both groups. Moreover, most of the strains were able to produce plant growth promoting molecules (ACC, IAA, NH3 and siderophores). Such ecological traits suggest that these new species of Burkholderia might be environmentally adaptable plant-associated bacteria and beneficial to plants.

Microbial species richness and assemblages across ultramafic ecosystems were investigated to assess the relationship between their distributional patterns and environmental traits. The structure of microorganism communities in the Koniambo massif, New Caledonia, was investigated using a metagenetic approach correlated with edaphic and floristic factors. Vegetation cover and soil properties significantly shaped the large phylogenetic distribution of operational taxonomic unit within microbial populations, with a mean per habitat of 3,477 (± 317) for bacteria and 712 (± 43) for fungi. Using variance partitioning, we showed that the effect of aboveground vegetation was the most significant descriptor for both bacterial and fungal communities. The floristic significant predictors explained 43% of the variation both for the bacterial and fungal community structures, while the edaphic significant predictors explained only 32% and 31% of these variations, respectively. These results confirm the previous hypothesis, that the distribution of microorganisms was more structured by the vegetation cover rather than the edaphic characteristics and that microbial diversity is not limited in ultramafic ecosystems.

Rhizosphere bacteria were isolated from Costularia spp., pioneer sedges from ultramafic soils in New Caledonia, which is a hotspot of biodiversity in the South Pacific. Genus identification, ability to tolerate edaphic constraints, and plant-growth-promoting (PGP) properties were analysed. We found that 105colony-forming units per gram of root were dominated by Proteobacteria (69%) and comprised 21 genera, including Burkholderia (28%), Curtobacterium (15%), Bradyrhizobium (9%), Sphingomonas (8%), Rhizobium (7%), and Bacillus (5%). High proportions of bacteria tolerated many elements of the extreme edaphic conditions: 82% tolerated 100 μmol·L–1chromium, 70% 1 mmol·L–1nickel, 63% 10 mmol·L–1manganese, 24% 1 mmol·L–1cobalt, and 42% an unbalanced calcium/magnesium ratio (1/16). These strains also exhibited multiple PGP properties, including the ability to produce ammonia (65%), indole-3-acetic acid (60%), siderophores (52%), and 1-aminocyclopropane-1-carboxylate (ACC) deaminase (39%); as well as the capacity to solubilize phosphates (19%). The best-performing strains were inoculated with Sorghum sp. grown on ultramafic substrate. Three strains significantly enhanced the shoot biomass by up to 33%. The most successful strains influenced plant nutrition through the mobilization of metals in roots and a reduction of metal transfer to shoots. These results suggest a key role of these bacteria in plant growth, nutrition, and adaptation to the ultramafic constraints.

The fungus Pisolithus albus forms ectomycorrhizal (ECM) associations with plants growing on extreme ultramafic soils, which are naturally rich in heavy metals such as nickel. Both nickel-tolerant and nickel-sensitive isolates of P. albus are found in ultramafic soils in New Caledonia, a biodiversity hotspot in the Southwest Pacific. The aim of this work was to monitor the expression of genes involved in the specific molecular response to nickel in a nickel-tolerant P. albus isolate. We used pyrosequencing and quantitative polymerase chain reaction (qPCR) approaches to investigate and compare the transcriptomes of the nickel-tolerant isolate MD06-337 in the presence and absence of nickel. A total of 1,071,375 sequencing reads were assembled to infer expression patterns of 19,518 putative genes. Comparison of expression levels revealed that 30% of the identified genes were modulated by nickel treatment. The genes, for which expression was induced most markedly by nickel, encoded products that were putatively involved in a variety of biological functions, such as the modification of cellular components (53%), regulation of biological processes (27%) and molecular functions (20%). The 10 genes that pyrosequencing analysis indicated were induced the most by nickel were characterized further by qPCR analysis of both nickel-tolerant and nickel-sensitive P. albus isolates. Five of these genes were expressed exclusively in nickel-tolerant isolates as well as in ECM samples in situ, which identified them as potential biomarkers for nickel tolerance in this species. These results clearly suggest a positive transcriptomic response of the fungus to nickel-rich environments. The presence of both nickel-tolerant and nickel-sensitive fungal phenotypes in ultramafic soils might reflect environment-dependent phenotypic responses to variations in the effective concentrations of nickel in heterogeneous ultramafic habitats.

International audience; In New Caledonia, a hot spot of biodiversity, plants from the Cyperaceae family are mostly endemic and considered pioneers of the nickel-rich natural serpentine ecosystem. The aim of the study was to highlight the mycorrhizal status of these Cyperaceae and to bring new insights into the role of this symbiosis in plant tolerance to ultramafic soils. Nine Cyperaceae species were studied and presented evidence of root colonization by arbuscular mycorrhizas (AMs), with frequencies ranging from 8% to 57%. The highest level of AM colonization was observed in plants from the endemic dominant genus Costularia. Molecular evidence demonstrated the presence of Glomus species inside the roots. In a controlled greenhouse assay, AM inoculation of Costularia comosa grown under ultramafic conditions significantly enhanced plant growth, with an increase in biomass by up to 2.4-fold for shoots and 1.2-fold for roots, and also reduced nickel content in roots by 2.5-fold, as compared with the controls. All these data support our hypotheses (i) that a relationship exists between the mycorrhizal status of Cyperaceae and their habitat, and (ii) that AM have a positive role in plant tolerance to ultramafic soils (mineral nutrition and metal tolerance), suggesting the use of these pioneer plants with AM management as potential tools for nickel mine site rehabilitation in New Caledonia.; En Nouvelle-Calédonie, un point chaud de la biodiversité, les plantes appartenant à la famille des Cypéracées sont principalement endémiques et sont considérées comme pionnières de l’écosystème naturel de la serpentine riche en nickel. Cette étude visait à éclaircir le statut mycorhizien de ces Cypéracées et d’apporter un nouvel aperçu de rôle de cette symbiose dans la tolérance des végétaux aux sols ultrabasiques. Neuf espèces de Cypéracées ont été étudiées et la colonisation des racines par des mycorhizes arbusculaires (MA) a été mise en évidence avec des fréquences variant de 8 % à 57 %. Le niveau de MA le plus élevé a été noté chez les plantes du genre endémique dominant Costularia. Les données moléculaires ont démontré la présence de Glomus sp. à l’intérieur des racines. Lors d’un essai en serre contrôlée, l’inoculation de MA chez Costularia comosa cultivée en conditions ultrabasiques a significativement accru la croissance des plants, avec une augmentation de la biomasse des pousses de 2,4 fois et des racines de 1,2 fois, réduisant aussi le contenu en nickel des racines de 2,5 fois, tout cela par rapport au contrôle. Toutes ces données plaident en faveur de l’hypothèse (i) d’une relation entre le statut en mycorhize des Cypéracées et leur habitat, et (ii) du rôle positif des MA dans la tolérance aux sols ultrabasiques (nutrition minérale et tolérance aux métaux), suggérant que ces plantes pionnières parallèlement à la gestion des MA puissent constituer des outils potentiels de réhabilitation des sites miniers de nickel en Nouvelle-Calédonie.

International audience; Ectomycorrhizal (ECM) Pisolithus albus (Cooke & Massee), belonging to the ultramafic ecotype isolated in nickel-rich serpentine soils from New Caledonia (a tropical hotspot of biodiversity) and showing in vitro adaptive nickel tolerance, were inoculated to Eucalyptus globulus Labill used as a Myrtaceae plant-host model to study ectomycorrhizal symbiosis. Plants were then exposed to a nickel (Ni) dose-response experiment with increased Ni treatments up to 60 mg kg (- 1) soil as extractable Ni content in serpentine soils. Results showed that plants inoculated with ultramafic ECM P. albus were able to tolerate high and toxic concentrations of Ni (up to 60 mu g g (- 1)) while uninoculated controls were not. At the highest Ni concentration tested, root growth was more than 20-fold higher and shoot growth more than 30-fold higher in ECM plants compared with control plants. The improved growth in ECM plants was associated with a 2.4-fold reduction in root Ni concentration but a massive 60-fold reduction in transfer of Ni from root to shoots. In vitro, P. albus strains could withstand high Ni concentrations but accumulated very little Ni in its tissue. The lower Ni uptake by mycorrhizal plants could not be explained by increased release of metal-complexing chelates since these were 5- to 12-fold lower in mycorrhizal plants at high Ni concentrations. It is proposed that the fungal sheath covering the plant roots acts as an effective barrier to limit transfer of Ni from soil into the root tissue. The degree of tolerance conferred by the ultramafic P. albus isolates to growth of the host tree species is considerably greater than previously reported for other ECM. The primary mechanisms underlying this improved growth were identified as reduced Ni uptake into the roots and markedly reduced transfer from root to shoot in mycorrhizal plants. The fact that these positive responses were observed at Ni concentrations commonly observed in serpentinic soils suggests that ultramafic ecotypes of P. albus could play an important role in the adaptation of tree species to soils containing high concentrations of heavy metals and aid in strategies for ecological restoration.

Contact: philippe.jourand@ird.fr; International audience; Isolates of ectomycorrhizal Pisolithus albus were sampled from both ultramafic and volcano-sedimentary soils in New Caledonia, a tropical hotspot of biodiversity, to investigate the relationships between genetic diversity and edaphic constraint through tolerance to nickel (Ni). Carpophore description, spore morphology and phylogenetic analysis based on internal transcribed spacer (ITS) rDNA sequences confirmed that all isolates belong to P. albus and are closely related to other Australasian specimens. Using molecular tools, ITS-restriction fragment length polymorphism and amplified fragment length polymorphism markers, we showed the existence of two distinct genetic clusters within P. albus: ultramafic and volcano-sedimentary. Mycelia response to Ni toxicity supports such a population structure. Pisolithus albus from ultramafic soils included isolates with a high diversity of in vitro Ni tolerance, with both Ni-tolerant isolates (average Ni EC(50) at 575 mu M) and Ni-sensitive isolates (average Ni EC(50) at 37 mu M). In contrast, all isolates from volcano-sedimentary soils were found to be Ni sensitive (average Ni EC(50) at 32 mu M). We highlight that (1) P. albus population from ultramafic soils of New Caledonia are genetically structured in ecotype, and that (2) Ni tolerance among ultramafic isolates suggests an adaptive physiological response to Ni toxicity.

Acacia spirorbis is a leguminous tree from New Caledonia naturally found on a wide range of soils (calcareous, ultramafic and volcano-sedimentary) and able to establish symbioses with soil microorganisms, including nitrogen-fixing bacteria. The contribution of this symbiosis in the plant adaptation to contrasted edaphic environments, sometimes extremes, has been investigated according to i) the nitrogen-fixing potential of A. spirorbis in its natural ecosystems, ii) the characterization of its rhizobia and iii) the plant adaptive response to heavy metals inside its roots tissues, especially in nodules.Therefore, we revealed that the nitrogen-fixing symbiosis provided more than 80% of the plant total nitrogen in natural population naturally occurring on calcareous, ultramafic and volcano-sedimentary soils. This value is top notch among other Acacia species, where A. mangium, A. melanoxylon and A. mucronata presented mean values of 50%, 43% and 58%, respectively. Acacia spirorbis established nitrogen-fixing symbioses with alpha- and bêta-proteobacteria, genus Bradyrhizobium and Paraburkholderia, respectively, thus revealing a wide range of symbiotic partner and a low selectivity. Noteworthy, the taxonomy and phenotypes of these symbionts are structured and adapted to edaphic parameters. Finally, chemical signatures of internal nodules tissues presented similarities with soils chemical properties, thus indicating a potential management of heavy metals inside these tissues.Altogether, these data suggest that the nitrogen-fixing symbiosis might significantly contributes to Acacia spirorbis adaptation towards contrasted soils with strong edaphic conditions as an extreme polymetallic toxicity.; Acacia spirorbis est une légumineuse endémique de Nouvelle-Calédonie se développant sur des sols calcaires, métallifères et volcano-sédimentaires, établissant des symbioses avec des bactéries fixatrices d’azote. Pour comprendre la contribution de la symbiose dans l’adaptation de la plante à des milieux contrastés et parfois extrêmes, nous avons évalué la fixation d’azote en conditions naturelles, caractérisé les rhizobia associés à cette plante et analysé la réponse adaptative de la plante aux éléments traces métalliques dans ses tissus racinaires, notamment au niveau des nodules.Nous avons mis en évidence que la symbiose rhizobienne fournissait plus de 80% de l’azote total chez des populations naturelles d’A. spirorbis se développant sur des sites d’études présentant des sols calcaires, métallifères et volcano-sédimentaires. Cette valeur est remarquable puisque chez A. mangium, A. melanoxylon et A. mucronata, les valeurs moyennes sont respectivement de 50%, 43% et 58%. Les rhizobia symbiotiques associés à A. spirorbis appartiennent aux alpha- et bêta-protéobactéries, genres Bradyrhizobium et Paraburkholderia, révélant ainsi une très large gamme de symbiontes et une faible sélectivité de partenaire. De manière remarquable, la taxonomie et la phénotypie de ces souches sont structurées et adaptées aux conditions édaphiques. Enfin, les signatures chimiques des tissus internes des nodules reflètent les propriétés chimiques des sols dans lesquels ils se sont développés, indiquant une potentielle gestion des éléments traces métalliques dans ces tissus.Tous ces éléments suggèrent que la symbiose fixatrice d’azote contribue de manière significative dans l’adaptation d’Acacia spirorbis à des sols contrastés et pouvant présenter une toxicité polymétallique extrême.