Seed endophytes and rhizosphere microbiome of Imperata cylindrica, a pioneer plant of abandoned mine lands.

Some plant-associated microorganisms could improve host plants biotic and abiotic stress tolerance. Imperata cylindrica is a dominant pioneer plant in some abandoned mine lands with higher concentrations of heavy metal (HM). To discover the specific microbiome of I. cylindrica in this extreme environment and evaluate its role, the microbiome of I. cylindrica's seeds and rhizosphere soils from HM heavily contaminated (H) and lightly contaminated (L) sites were studied. It was found that HM-contamination significantly reduced the richness of endophytic bacteria in seeds, but increased the abundance of resistant species, such as Massilia sp. and Duganella sp. Spearman's rank correlation coefficient analysis showed that both Massilia sp. and Duganella sp. showed a significant positive correlation with Zn concentration, indicating that it may have a strong tolerance to Zn. A comparison of the microbiome of rhizosphere soils (RS) and adjacent bare soils (BS) of site H showed that I. cylindrica colonization significantly increased the diversity of fungi in rhizosphere soil and the abundance of Ascomycota associated with soil nutrient cycling. Spearman's rank correlation coefficient analysis showed that Ascomycota was positively correlated with the total nitrogen. Combined with the fact that the total nitrogen content of RS was significantly higher than that of BS, we suppose that Ascomycota may enhance the nitrogen fixation of I. cylindrica, thereby promoting its growth in such an extreme environment. In conclusion, the concentration of HM and nutrient contents in the soil significantly affected the microbial community of rhizosphere soils and seeds of I. cylindrica, in turn, the different microbiomes further affected soil HM concentration and nutrient contents. The survival of I. cylindrica in HM severely contaminated environment may mainly be through recruiting more microorganisms that can enhance its nutrition supply.

Pioneer plants enhance soil multifunctionality by reshaping underground multitrophic community during natural succession of an abandoned rare earth mine tailing.

Spontaneous natural succession in metal mine tailings is fundamental to the rehabilitation of bare tailing. Here, an abandoned rare earth element (REE) mine tailing with spontaneous colonisation by pioneer plants with different functional traits was selected. Soil nutrient cycling, fertility, organic matter decomposition as well as underground organismal communities and their multitrophic networks were investigated. Compared with the bare tailing, the colonisation with Lycopodium japonicum, Miscanthus sinensis, and Dicranopteris dichotoma increased soil multifunction by 222%, 293%, and 525%, respectively. This was accompanied by significant changes in soil bacterial and protistan community composition and increased soil multitrophic network complexity. Rhizospheres of different plant species showed distinct microbial community composition compared to that of bare tailing. Some WPS-2, Chloroflexi, and Chlorophyta were mainly present in the bare tailing, while some Proteobacteria and Cercozoa were predominantly seen in the rhizosphere. Pearson correlation and Random Forest revealed the biotic factors driving soil multifunction. Structural equation modelling further revealed that pioneer plants improved soil multifunction primarily by decreasing the microbial biodiversity and increasing the multitrophic network complexity. Overall, this highlights the importance of subterrestrial organisms in accelerating soil rehabilitation during natural succession and provides options for the ecological restoration of degraded REE mining areas.

A Nitrate-Transforming Bacterial Community Dominates in the Miscanthus Rhizosphere on Nitrogen-Deficient Volcanic Deposits of Miyake-jima.

The perennial gramineous grass Miscanthus condensatus functions as a major pioneer plant in colonizing acidic volcanic deposits on Miyake-jima, Japan, despite a lack of nitrogen nutrients. The nitrogen cycle in the rhizosphere is important for the vigorous growth of M. condensatus in this unfavorable environment. In the present study, we identified the nitrogen-cycling bacterial community in the M. condensatus rhizosphere on these volcanic deposits using a combination of metagenomics and culture-based analyses. Our results showed a large number of functional genes related to denitrification and dissimilatory nitrate reduction to ammonium (DNRA) in the rhizosphere, indicating that nitrate-transforming bacteria dominated the rhizosphere biome. Furthermore, nitrite reductase genes (i.e., nirK and nirS) related to the denitrification and those genes related to DNRA (i.e., nirB and nrfA) were mainly annotated to the classes Alpha-proteobacteria, Beta-proteobacteria, and Gamma-proteobacteria. A total of 304 nitrate-succinate-stimulated isolates were obtained from the M. condensatus rhizosphere and were classified into 34 operational taxonomic units according to amplified 16S rRNA gene restriction fragment pattern analysis. Additionally, two strains belonging to the genus Cupriavidus in the class Beta-proteobacteria showed a high in vitro denitrifying activity; however, metagenomic results indicated that the DNRA-related rhizobacteria appeared to take a major role in the nitrogen cycle of the M. condensatus rhizosphere in recent Miyake-jima volcanic deposits. This study elucidates the association between the Miscanthus rhizosphere and the nitrate-reducing bacterial community on newly placed volcanic deposits, which furthers our understanding of the transformation of nitrogen nutrition involved in the early development of vegetation.

Vanadium accumulation mode of Heteropogon contortus and its driving factors in Majiatian tailing reservoir in Panzhihua, Southwestern China.

Native plants in metal pollution sites have great potentials for mine rehabilitation. In the presented work, we investigated Vanadium (V) concentrations of soils and plants (Heteropogon contortus) in Majiatian V-Ti magnetite tailing reservoir in Panzhihua, Southwestern China. The objectives were to explore the V accumulation mode of H. contortus and its driving factors, as well as the phytoremediation potential of H. contortus. As the results, H. contortus accumulated 37.53 mg/kg and 8.69 mg/kg of V in root and aerial part, respectively. With the increase of rehabilitation age, root V concentrations decreased, while aerial part V concentrations remained constant. The significant negative correlations between root V and soil V, acid-soluble V (VHAc) (P < 0.05) indicated that increasing soil V and VHAc concentrations drove the V accumulation mode of H. contortus. Soil properties had a little influence on the V accumulation mode of H. contortus. Therefore, H. contortus might be not the suitable plant extractant to remove V from mine tailing for its lower V accumulation capacity. On the other hand, it can tolerate high V stress through elimination and detoxification/isolation V. Furthermore, the settlement of H. contortus increased the content of soil organic matter and might thus improve the soil quality. The cover of H. contortus is also beneficial to reduce the dispersion of the tailings and prevent contaminating surrounding soil. Therefor it showed a great potential to serve as a pioneer plant in the remediation of V-rich tailing reservoirs and other V-contaminated sites with similar poor soil condition.

[Acid Mine Wasteland Reclamation by Juncus ochraceus Buchen as a Potential Pioneer Plant].

Ecological reclamation is the major method for the revegetation of acid mine wasteland worldwide. In this study the pH, fertility characteristics, and heavy metal content of soils from Laili Mountain mine wasteland were analyzed. The research also studied the morphological characteristics and the heavy metal in Juncus ochraceus Buchen as well as its resistance to acid, adaptability to soil fertility in abandoned land, and tolerance to heavy metal pollution (e.g., Zn and Cu) to determine its remediation potential as a pioneer plant for acid mine wasteland. Results showed that the pH of soils in the study area were acidic, ranging from 3.46 to 4.01.The contents of organic matter, total potassium, total phosphorus, and available phosphorus was poor, being 10.28-25.75 g·kg-1, 8.84-9.32 g·kg-1, 0.56-0.63 g·kg-1, and 1.82-5.72 mg·kg-1 respectively.The contents of Zn, Cu, and Fe in the soil ranged between 54.93 and 114.49 mg·kg-1, 92.53 and 127.59 mg·kg-1, and 47133.60 and 112259.63 mg·kg-1, whereby the Cu content was 1.85-2.55 times higher than the risk screening value. The height of Juncus ochraceus Buchen in the study area ranged from 43.77 cm to 55.42 cm, which was shorter than the average plant height of the control group (51.38-57.66 cm); however, this was not a significant difference, thus indicating that this plant was resistant to acidic soil and heavy metal pollution.Further analysis showed that Juncus ochraceus Buchen had accumulating capacity for both Cu and Zn, and transferring capacity for Zn as well. Thus, it had potential in heavy metal accumulation and absorption.Plant height was significantly related to the available phosphorus content in the rhizosphere soil. In the future, soil fertility could be improved by supplementing fertilizers containing available phosphorus when Juncus ochraceus Buchen is planted on the land of abandoned mines as a pioneer plant. Comprehensive analysis revealed that Juncus ochraceus Buchen had great potential as a pioneer plant to remediate acid mine wasteland.

Tissue-Specific Dynamics in the Endophytic Bacterial Communities in Arctic Pioneer Plant Oxyria digyna.

The rapid developments in the next-generation sequencing methods in the recent years have provided a wealth of information on the community structures and functions of endophytic bacteria. However, the assembly processes of these communities in different plant tissues are still currently poorly understood, especially in wild plants in natural settings. The aim of this study was to compare the composition of endophytic bacterial communities in leaves and roots of arcto-alpine pioneer plant Oxyria digyna, and investigate, how plant tissue (leaf or root) or plant origin affect the community assembly. To address this, we planted micropropagated O. digyna plants with low bacterial load (bait plants) in experimental site with native O. digyna population, in the Low Arctic. The endophytic bacterial community structures in the leaves and roots of the bait plants were analyzed after one growing season and one year in the field, and compared to those of the wild plants growing at the same site. 16S rRNA gene targeted sequencing revealed that endophytic communities in the roots were more diverse than in the leaves, and the diversity in the bait plants increased in the field, and was highest in the wild plants. Both tissue type and plant group had strong impact on the endophytic bacterial community structures. Firmicutes were highly abundant in the leaf communities of both plant types. Proteobacteria and Bacteroidetes were more abundant in the roots, albeit with different relative abundances in different plant groups. The community structures in the bait plants changed in the field over time, and increasingly resembled the wild plant endophytic communities. This was due to the changes in the relative abundances of several bacterial taxa, as well as species acquisition in the field, but with no species turnover. Several OTUs that were acquired by the bait plants in the field and represent phosphate solubilizing and diazotrophic bacterial taxa, suggesting major role in nutrient acquisition of these bacteria for this nonmycorrhizal plant, thriving in the nutrient poor arctic soils.

Topsoil structure stability in a restored floodplain: Impacts of fluctuating water levels, soil parameters and ecosystem engineers.

Ecosystem services provided by floodplains are strongly controlled by the structural stability of soils. The development of a stable structure in floodplain soils is affected by a complex and poorly understood interplay of hydrological, physico-chemical and biological processes. This paper aims at analysing relations between fluctuating groundwater levels, soil physico-chemical and biological parameters on soil structure stability in a restored floodplain. Water level fluctuations in the soil are modelled using a numerical surface-water-groundwater flow model and correlated to soil physico-chemical parameters and abundances of plants and earthworms. Causal relations and multiple interactions between the investigated parameters are tested through structural equation modelling (SEM). Fluctuating water levels in the soil did not directly affect the topsoil structure stability, but indirectly through affecting plant roots and soil parameters that in turn determine topsoil structure stability. These relations remain significant for mean annual days of complete and partial (>25%) water saturation. Ecosystem functioning of a restored floodplain might already be affected by the fluctuation of groundwater levels alone, and not only through complete flooding by surface water during a flood period. Surprisingly, abundances of earthworms did not show any relation to other variables in the SEM. These findings emphasise that earthworms have efficiently adapted to periodic stress and harsh environmental conditions. Variability of the topsoil structure stability is thus stronger driven by the influence of fluctuating water levels on plants than by the abundance of earthworms. This knowledge about the functional network of soil engineering organisms, soil parameters and fluctuating water levels and how they affect soil structural stability is of fundamental importance to define management strategies of near-natural or restored floodplains in the future.

Restoration with pioneer plants changes soil properties and remodels the diversity and structure of bacterial communities in rhizosphere and bulk soil of copper mine tailings in Jiangxi Province, China.

To unravel the ecological function played by pioneer plants in the practical restoration of mine tailings, it is vital to explore changes of soil characteristics and microbial communities in rhizosphere and bulk soil following the adaptation and survival of plants. In the present study, the diversity and structure of rhizospheric bacterial communities of three pioneer plants in copper mine tailings were investigated by Illumina MiSeq sequencing, and the effects of pioneer plants on soil properties were also evaluated. Significant soil improvement was detected in rhizospheric samples, and Alnus cremastogyne showed higher total organic matter, total nitrogen, and available phosphorus than two other herbaceous plants. Microbial diversity indices in rhizosphere and bulk soil of reclaimed tailings were significantly higher than bare tailings, even the soil properties of bulk soil in reclaimed tailings were not significantly different from those of bare tailings. A detailed taxonomic composition analysis demonstrated that Alphaproteobacteria and Deltaproteobacteria, Chloroflexi, Acidobacteria, and Gemmatimonadetes showed significantly higher relative abundance in rhizosphere and bulk soil. In contrast, Gammaproteobacteria and Firmicutes were abundant in bare tailings, in which Bacillus, Pseudomonas, and Lactococcus made up the majority of the bacterial community (63.04%). Many species within known heavy metal resistance and nutrient regulatory microorganism were identified in reclaimed tailings, and were more abundant among rhizospheric microbes. Hierarchical clustering and principal coordinate analysis (PCoA) analysis demonstrated that the bacterial profiles in the rhizosphere clustered strictly together according to plant types, and were distinguishable from bulk soil. However, we also identified a large shared OTUs that occurred repeatedly and was unaffected by highly diverse soil properties in rhizosphere and bulk samples. Redundancy analysis indicated that water content and Cu and As concentrations were the main environmental regulators of microbial composition. These results suggest that the interactive effect of pioneer plants and harsh soil environmental conditions remodel the specific bacterial communities in rhizosphere and bulk soil in mine tailings. And A. cremastogyne might be approximate candidate for phytoremediation of mine tailings for better soil amelioration effect and relative higher diversity of bacterial community in rhizosphere.

Landscape metrics as functional traits in plants: perspectives from a glacier foreland.

Spatial patterns of vegetation arise from an interplay of functional traits, environmental characteristics and chance. The retreat of glaciers offers exposed substrates which are colonised by plants forming distinct patchy patterns. The aim of this study was to unravel whether patch-level landscape metrics of plants can be treated as functional traits. We sampled 46 plots, each 1 m × 1 m, distributed along a restricted range of terrain age and topsoil texture on the foreland of the Nardis glacier, located in the South-Eastern Alps, Italy. Nine quantitative functional traits were selected for 16 of the plant species present, and seven landscape metrics were measured to describe the spatial arrangement of the plant species' patches on the study plots, at a resolution of 1 cm × 1 cm. We studied the relationships among plant communities, landscape metrics, terrain age and topsoil texture. RLQ-analysis was used to examine trait-spatial configuration relationships. To assess the effect of terrain age and topsoil texture variation on trait performance, we applied a partial-RLQ analysis approach. Finally, we used the fourth-corner statistic to quantify and test relationships between traits, landscape metrics and RLQ axes. Floristically-defined relevé clusters differed significantly with regard to several landscape metrics. Diversity in patch types and size increased and patch size decreased with increasing canopy height, leaf size and weight. Moreover, more compact patch shapes were correlated with an increased capacity for the conservation of nutrients in leaves. Neither plant species composition nor any of the landscape metrics were found to differ amongst the three classes of terrain age or topsoil texture. We conclude that patch-level landscape metrics of plants can be treated as species-specific functional traits. We recommend that existing databases of functional traits should incorporate these type of data.

Frugivory and seed dispersal of Solanum granuloso-leprosum Dunal (Solanaceae) by birds in deciduous seasonal forest / Frugivoria e dispersão de sementes em Solanum granuloso-leprosum Dunal (Solanaceae) por aves em Floresta Estacional Decidual

Abstract The goal of this study was to identify which bird species consume Solanum granuloso-leprosum fruits and disperse its seeds. 60 hours of focal observations were carried out between April and May 2006 on the edge of a deciduous forest fragment in the Uruguay River region, Rio Grande do Sul state, Brazil. Ten species were observed in total removing 443 fruits. Saltator similis removed 61.8% of the fruits, followed by Tangara sayaca (17.1%), Pipraeidea bonariensis (11.7%), and T. preciosa (6.8%), while the remaining six species accounted for only 2.5% of the fruits removed. Most fruit removal occurred early in the day or mid-afternoon. The most common feeding behaviors were picking (60.7%), followed by stalling (23%) and hovering (16%). Birds flew more than 10 m from the fruit plant in 62% of the removal events. All bird species observed here may be considered potential dispersers of S. granuloso-leprosum, as they moved the seeds away from the mother plant where strong competition and predation are likely to occur. Results also suggest that S. granuloso-leprosum may be useful in ecological restoration programs.

Resumo O objetivo deste estudo foi identificar quais as espécies de aves consomem frutos de Solanum granuloso-leposum e dispersam suas sementes. Para tanto, 60 horas de observações focais foram realizadas entre abril e maio de 2006 na borda de um fragmento de floresta decídua na região do rio Uruguai, Rio Grande do Sul, Brasil. Dez espécies foram observadas removendo 443 frutos. Saltator similis removeu 61,8% dos frutos, seguido de Tangara sayaca (17,1%), Pipraeidea bonariensis (11,7%), e T. preciosa (6,8%), enquanto as restantes seis espécies representaram apenas 2,5% dos frutos removidos. A maioria das remoções dos frutos ocorreram no início do dia ou meio da tarde. Os comportamentos alimentares mais comuns foram picking (60,7%), seguido por stalling (23%) e hovering (16%). As aves voaram mais de 10 m da planta de onde removeram frutos em 62% dos eventos de remoção. Todas as espécies de aves observadas aqui podem ser consideradas potenciais dispersores de S. granuloso-leprosum, moveram sementes para longe da planta-mãe, onde a concorrência e a predação são mais susceptíveis de ocorrer. Os resultados também sugerem que S. granuloso-leprosum pode ser útil em programas de restauração ecológica.

Anatomy, histochemistry and micromorphology of leaves of Solanum granuloso-leprosum dunal / Anatomia, histoquímica e micromorfologia de folhas de Solanum granuloso-leprosum dunal

In the present work the anatomical, histochemical and micromorphological features of S. granuloso-leprosum leaves were approached in order to evaluate its characteristics associated with its pioneer role. Glandular and non-glandular trichomes were observed on both epidermal surfaces, although in greater number on the ab axial surface. Stellate trichomes presented a thick lignified cell wall. Leaves were amphiestomatic with a single palisade layer and a slightly smaller spongy parenchyma. The epidermal cells of the abaxial surface were shorter than the adaxial ones, both with stomata paracytic. Vascular bundles were bicolateral and idioblasts with conspicuous crystalliferous inclusions were observed in the mesophyll. Lipid drops were evidenced in the spongy parenchyma by Sudan III, Nile Blue, Nadi reagent and Sudan Black histochemical tests. Negative results for alkaloids and phenol compounds were observed. The evaluated anatomical and hystochemical data highlights mesophytic characteristics in accordance with S. granuloso-leprosum pioneer plant role.

No presente trabalho foi abordada a anatomia, histoquímica e micromorfologia de folhas de S. granuloso-leprosum no intuito de se levantar características associadas a sua função de espécie pioneira. Tricomas glandulares e não glandulares foram observados em ambas as faces da epiderme, apesar de maior número na superfície abaxial. Tricomas estrelados apresentaram uma parede espessa e lignificada. As folhas são anfiestomáticas com uma única camada de paliçada e um parênquima lacunoso com células menores que as do paliçádico. As células da face abaxial da epiderme são menores, e ambas as faces apresentam estômatos paracíticos. Os feixes vasculares são bicolaterais e idioblastos com inclusões cristalinas conspícuas. Gotas de lipídeo foram evidenciadas no parênquima lacunoso com os testes histoquímicos de Sudan III, azul do Nilo, reagente de Nadi e Sudan Black. Foram observados resultados negativos para alcaloides e fenóis. Os dados anatômicos e histoquímicos avaliados permitiram verificar que S. granuloso-leprosum apresentam características mesófilas, os quais estão em conformidade com o seu papel fitossociológicos como pioneira.