Co-inoculation with a bacterium and arbuscular mycorrhizal fungi improves root colonization, plant mineral nutrition, and plant growth of a Cyperaceae plant in an ultramafic soil.

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.

New Caledonian ultramafic conditions structure the features of Curtobacterium citreum strains that play a role in plant adaptation.

The present study focused on the characterization of 10 Curtobacterium citreum strains isolated from the rhizosphere of pioneer plants growing on ultramafic soils from New Caledonia. Taxonomic status was investigated using a polyphasic approach. Three strains (BE, BB, and AM) were selected in terms of multiple-metal resistance and plant-growth-promoting traits. They were tested on sorghum growing on ultramafic soil and compared with the reference strain C. citreum DSM20528T. To better understand the bacterial mechanisms involved, biosorption, bioaccumulation, and biofilm formation were investigated for the representative strain of the ultramafic cluster (strain BE) versus C. citreum DSM20528T. The polyphasic approach confirmed that all native isolates belong to the same cluster and are C. citreum. The inoculation of sorghum with strains BE and BB significantly reduced Ni content in shoots compared with inoculation with C. citreum DSM20528T and control values. This result was related to the higher Ni tolerance of the ultramafic strains compared with C. citreum DSM20528T. Ni biosorption and bioaccumulation showed that BE exhibited a lower Ni content, which is explained by the ability of this strain to produce exopolysaccharides involved in Ni chelation. We suggested that ultramafic C. citreum strains are more adapted to this substrate than is C. citreum DSM20528T, and their features allow them to enhance plant metal tolerance.

Is a mixture of arbuscular mycorrhizal fungi better for plant growth than single-species inoculants?

Inoculation of arbuscular mycorrhizal fungi (AMF) as plant growth promoters has mostly been conducted using single-species inoculum. In this study, we investigated whether co-inoculation of different native AMF species induced an improvement of plant growth in an ultramafic soil. We analyzed the effects of six species of AMF from a New Caledonian ultramafic soil on plant growth and nutrition, using mono-inoculations and mixtures comprising different numbers of AMF species, in a greenhouse experiment. The endemic Metrosideros laurifolia was used as a host plant. Our results suggest that, when the plant faced multiple abiotic stress factors (nutrient deficiencies and high concentrations of different heavy metals), co-inoculation of AMF belonging to different families was more efficient than mono-inoculation in improving biomass, mineral nutrition, Ca/Mg ratio, and tolerance to heavy metals of plants in ultramafic soil. This performance suggested functional complementarity between distantly related AMF. Our findings will have important implications for restoration ecology and mycorrhizal biotechnology applied to ultramafic soils.

Arbuscular mycorrhizal fungi and sewage sludge enhance growth and adaptation of Metrosideros laurifolia on ultramafic soil in New Caledonia: A field experiment.

The ecological restauration of nickel mining-degraded areas in New Caledonia is strongly limited by soil mineral nutrient deficiencies, heavy metal toxicity and slow growth rate of the native plant species. The improvement of revegetation technics needs multidisciplinary knowledge. A field experiment with relocated topsoil was assessed to test plant inoculation with a mix of three selected arbuscular mycorrhizal fungi (AMF) combined with sewage sludge amendment of the soil. Metrosideros laurifolia seedlings, an endemic Myrtaceae, were inoculated with the mixed AMF isolates and grown in a nursery for 18 months before being planted. 528 days after plantation, the dry weight of inoculated plants was 4 times higher than non-inoculated ones. AMF inoculated plants growing in sewage sludge amended soil showed a dry weight more than seven times higher than control plants. These differences were positively correlated with mycorrhizal colonization. However at this stage, AMF inoculated isolates were reduced in roots of M. laurifolia and replaced by several AMF indigenous species. This AMF diversity was higher in inoculated plants and in non-amended plots. Inoculated plants were characterized by a better mineral nutrition, a higher Ca/Mg ratio and a lower heavy metal translocation. In conclusion, this study showed that AMF inoculation combined with sewage sludge amendment of soil can improve ecological restoration of ultramafic mine-degraded areas.

New method for the identification of arbuscular mycorrhizal fungi by proteomic-based biotyping of spores using MALDI-TOF-MS.

Arbuscular mycorrhizal fungi (AMF, Glomeromycota) are mutualistic symbionts associated with majority of land plants. These fungi play an important role in plant growth, but their taxonomic identification remains a challenge for academic research, culture collections and inoculum producers who need to certify their products. Identification of these fungi was traditionally performed based on their spore morphology. DNA sequence data have successfully been used to study the evolutionary relationships of AMF, develop molecular identification tools and assess their diversity in the environment. However, these methods require considerable expertise and are not well-adapted for "routine" quality control of culture collections and inoculum production. Here, we show that Matrix-Assisted Laser Desorption Ionisation Time of Flight Mass Spectrometry proteomic-based biotyping is a highly efficient approach for AMF identification. Nineteen isolates belonging to fourteen species, seven genera and five families were clearly differentiated by MALDI biotyping at the species level, and intraspecific differentiation was achieved for the majority. AMF identification by MALDI biotyping could be highly useful, not only for research but also in agricultural and environmental applications. Fast, accurate and inexpensive molecular mass determination and the possibility of automation make MALDI-TOF-MS a real alternative to conventional morphological and molecular methods for AMF identification.

Burkholderia novacaledonica sp. nov. and B. ultramafica sp. nov. isolated from roots of Costularia spp. pioneer plants of ultramafic soils in New Caledonia.

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.

Arbuscular mycorrhizal fungi from New Caledonian ultramafic soils improve tolerance to nickel of endemic plant species.

In order to improve knowledge about the role of arbuscular mycorrhizal fungi (AMF) in the tolerance to heavy metals in ultramafic soils, the present study investigated the influence of two Glomus etunicatum isolates from New Caledonian ultramafic maquis (shrubland), on nickel tolerance of a model plant species Sorghum vulgare, and of two ultramafic endemic plant species, Alphitonia neocaledonica and Cloezia artensis. In a first step, plants were grown in a greenhouse, on sand with defined concentrations of Ni, to appreciate the effects of the two isolates on the alleviation of Ni toxicity in controlled conditions. In a second step, the influence of the AMF on A. neocaledonica and C. artensis plants grown in a New Caledonian ultramafic soil rich in extractable nickel was investigated. Ni reduced mycorrhizal colonization and sporulation of the fungal isolates, but the symbionts increased plant growth and adaptation of endemic plant species to ultramafic conditions. One of the two G. etunicatum isolates showed a stronger positive effect on plant biomass and phosphorus uptake, and a greater reduction in toxicity symptoms and Ni concentration in roots and shoots. The symbionts seemed to act as a barrier to the absorption of Ni by the plant and reduced root-to-shoot Ni translocation. Results indicate the potential of selected native AMF isolates from ultramafic areas for ecological restoration of such degraded ecosystems.

Synteny comparison between apes and human using fine-mapping of the genome.

Comparing the genomes of the great apes and human should provide novel information concerning the origins of humankind. Relative to the great apes, the human karyotype has one fewer chromosome pair, as human chromosome 2 derived from the telomeric fusion of two ancestral primate chromosomes. To identify the genomic rearrangements that accompanied human speciation, we initiated a comparative study between human, chimpanzee, and gorilla. Using the HAPPY mapping method, an acellular adaptation of the radiation hybrid method, we mapped a few hundred markers on the human, chimpanzee, and gorilla genomes. This allowed us to identify several chromosome rearrangements, in particular a pericentric inversion and a translocation. We precisely localized the synteny breakpoint that led to the formation of human chromosome 2. This breakpoint was confirmed by FISH mapping.