Kevin K. Newsham

A meta‐analysis of plant responses to dark septate root endophytes

• Dark septate endophytes (DSE) frequently colonize roots in the natural environment, but the effects of these fungi on plants are obscure, with previous studies indicating negative, neutral or positive effects on plant performance. • In order to reach a consensus for how DSE influence plant performance, meta-analyses were performed on data from 18 research articles, in which plants had been inoculated with DSE in sterile substrates. • Negative effects of DSE on plant performance were not recorded. Positive effects were identified on total, shoot and root biomass, and on shoot nitrogen (N) and phosphorus contents, with increases of 26-103% in these parameters for plants inoculated with DSE, relative to uninoculated controls. Inoculation increased total, shoot and root biomass by 52-138% when plants had not been supplied with additional inorganic N, or when all, or the majority, of N was supplied in organic form. Inoculation with the DSE Phialocephala fortinii was found to increase shoot and root biomass, shoot P concentration and shoot N content by 44-116%, relative to uninoculated controls. • The analyses here suggest that DSE enhance plant performance under controlled conditions, particularly when all, or the majority, of N is available in organic form.

Soil organic nitrogen mineralization across a global latitudinal gradient

[1] Understanding and accurately predicting the fate of carbon and nitrogen in the terrestrial biosphere remains a central goal in ecosystem science. Amino acids represent a key pool of C and N in soil, and their availability to plants and microorganisms has been implicated as a major driver in regulating ecosystem functioning. Because of potential differences in biological diversity and litter quality, it has been thought that soils from different latitudes and plant communities may possess intrinsically different capacities to perform key functions such as the turnover of amino acids. In this study we measured the soil solution concentration and microbial mineralization of amino acids in soils collected from 40 latitudinal points from the Arctic through to Antarctica. Our results showed that soil solution amino acid concentrations were relatively similar between sites and not strongly related to latitude. In addition, when constraints of temperature and moisture were removed, we demonstrate that soils worldwide possess a similar innate capacity to rapidly mineralize amino acids. Similarly, we show that the internal partitioning of amino acid-C into catabolic and anabolic processes is conservative in microbial communities and independent of global position. This supports the view that the conversion of high molecular weight (MW) organic matter to low MW compounds is the rate limiting step in organic matter breakdown in most ecosystems.

Acquisition and assimilation of nitrogen as peptide-bound and D-enantiomers of amino acids by wheat.

Nitrogen is a key regulator of primary productivity in many terrestrial ecosystems. Historically, only inorganic N (NH4 + and NO3 -) and L-amino acids have been considered to be important to the N nutrition of terrestrial plants. However, amino acids are also present in soil as small peptides and in D-enantiomeric form. We compared the uptake and assimilation of N as free amino acid and short homopeptide in both L- and D-enantiomeric forms. Sterile roots of wheat (Triticum aestivum L.) plants were exposed to solutions containing either 14C-labelled L-alanine, D-alanine, L-trialanine or D-trialanine at a concentration likely to be found in soil solution (10 µM). Over 5 h, plants took up L-alanine, D-alanine and L-trialanine at rates of 0.9±0.3, 0.3±0.06 and 0.3±0.04 µmol g−1 root DW h−1, respectively. The rate of N uptake as L-trialanine was the same as that as L-alanine. Plants lost ca.60% of amino acid C taken up in respiration, regardless of the enantiomeric form, but more (ca.80%) of the L-trialanine C than amino acid C was respired. When supplied in solutions of mixed N form, N uptake as D-alanine was ca.5-fold faster than as NO3 -, but slower than as L-alanine, L-trialanine and NH4 +. Plants showed a limited capacity to take up D-trialanine (0.04±0.03 µmol g−1 root DW h−1), but did not appear to be able to metabolise it. We conclude that wheat is able to utilise L-peptide and D-amino acid N at rates comparable to those of N forms of acknowledged importance, namely L-amino acids and inorganic N. This is true even when solutes are supplied at realistic soil concentrations and when other forms of N are available. We suggest that it may be necessary to reconsider which forms of soil N are important in the terrestrial N cycle.

Metagenomic analysis of a southern maritime Antarctic soil

Our current understanding of Antarctic soils is derived from direct culture on selective media, biodiversity studies based on clone library construction and analysis, quantitative PCR amplification of specific gene sequences and the application of generic microarrays for microbial community analysis. Here, we investigated the biodiversity and functional potential of a soil community at Mars Oasis on Alexander Island in the southern Maritime Antarctic, by applying 454 pyrosequencing technology to a metagenomic library constructed from soil genomic DNA. The results suggest that the commonly cited range of phylotypes used in clone library construction and analysis of 78–730 OTUs (de-replicated to 30–140) provides low coverage of the major groups present (∼5%). The vast majority of functional genes (>77%) were for structure, carbohydrate metabolism, and DNA/RNA processing and modification. This study suggests that prokaryotic diversity in Antarctic terrestrial environments appears to be limited at the generic level, with Proteobacteria, Actinobacteria being common. Cyanobacteria were surprisingly under-represented at 3.4% of sequences, although ∼1% of the genes identified were involved in CO2 fixation. At the sequence level there appeared to be much greater heterogeneity, and this might be due to high divergence within the relatively restricted lineages which have successfully colonized Antarctic terrestrial environments.

Root-Fungal Associations of Colobanthus quitensis and Deschampsia antarctica in the Maritime and Subantarctic

ABSTRACT The two native Antarctic vascular plant species, Colobanthus quitensis and Deschampsia antarctica, were sampled from 15 points along a 1480 km latitudinal transect from South Georgia (54°S, 36°W) through to the Léonie Islands on the western Antarctic Peninsula (67°S, 68°W). Roots of plants were cleared and stained and fungal structures recorded. The commonest type of fungal association was that formed by dark septate endophytes (DSE): 32% and 27% of the root lengths of C. quitensis and D. antarctica were colonized by hyphae of these fungi, respectively. Hyaline and stained septate hyphae were also common in roots. Coarse and fine arbuscular mycorrhiza (AM) occurred in the roots of both plant species from South Georgia, and fine AM colonization with occasional arbuscules was also sporadically recorded in roots from the South Shetland Islands, suggesting functional associations between higher plants and AM symbionts. Fungal abundances were not associated with soil chemistry, but AM abundance was associated with seasonal surface air temperature, with lower colonization in more southerly, colder habitats. We conclude that DSE are widespread, and that AM fungi are sparse but present and decline in abundance at higher latitudes, in the roots of C. quitensis and D. antarctica.

Minimal influence of water and nutrient content on the bacterial community composition of a maritime antarctic soil.

Bacterial community composition was determined by culture-independent PCR-based methods in two soils differing markedly in their water, C, N and P contents sampled from Mars Oasis on Alexander Island, western Antarctic Peninsula. 16S rRNA sequences of the phyla Actinobacteria, Cyanobacteria, α-Proteobacteria and Acidobacteria were commonly (> 8% frequency) obtained from soil. Those of β-, γ- and δ-Proteobacteria, Chloroflexi, Bacteroidetes, Verrucomicrobia, Planctomycetes, Gemmatimonadetes and Firmicutes were less frequent. Comparisons of slopes of collectors curves and the Shannon-Weiner diversity index indicated no difference in overall bacterial diversity between the two soils, although sequences of δ-Proteobacteria and the cyanobacterial genus Leptolyngbya were more commonly derived from the soil with the higher water and nutrient content. The data suggest that different levels of soil water, C, N and P have only a minor effect on the bacterial community composition of maritime Antarctic soils.

The biology and ecology of the liverwort Cephaloziella varians in Antarctica

Abstract The biology and ecology of Cephaloziella varians, the most widespread and abundant liverwort in Antarctica, are reviewed. A description of the species is given, together with information on its geographical distribution, reproduction, habitats, associated organisms and responses to environmental stresses. Characteristics of its photosynthetic physiology are also presented, including data on oxygen evolution rates and chlorophyll a fluorescence parameters. Substratum and tissue chemistry, water relations and pigments are discussed, along with recent data demonstrating that the dark pigment in the apical leaves of C. varians is the anthocyanidin riccionidin A. Recent studies showing that the ericoid mycorrhizal symbiont Rhizoscyphus ericae is present in the tissues of the plant at a wide range of locations in the maritime and sub-Antarctic are also described. It is evident, from the literature reviewed, that C. varians has several adaptations that enable it to survive in the Antarctic biome, explaining its survival at higher latitudes than any other hepatic. The species’ major adaptations include the synthesis of riccionidin A in apical leaves, enabling efficient heat absorption and protection from photoinhibition, and the presence in stems and rhizoids of fungal hyphae, which are potentially beneficial to the hepatic’s nutrition and possibly also synthesize cryoprotectants.

Vegetation cover regulates the quantity, quality and temporal dynamics of dissolved organic carbon and nitrogen in Antarctic soils.

Populations of the two native Antarctic vascular plant species (Deschampsia antarctica and Colobanthus quitensis) have expanded rapidly in recent decades, yet little is known about the effects of these expansions on soil nutrient cycling. We measured the concentrations of dissolved organic carbon (DOC) and nitrogen (DON), amino acids and inorganic N in soils under these two vascular plant species, and under mosses and lichens, over a growing season at Signy Island in the maritime Antarctic. We recorded higher concentrations of nitrate, total dissolved nitrogen, DOC, DON and free amino acids in soil under D. antarctica and C. quitensis than in lichen or moss dominated soils. Each vegetation cover gave a unique profile of individual free amino acids in soil solution. Significant interactions between soil type and time were found for free amino acid concentrations and C/N ratios, indicating that vascular plants significantly change the temporal dynamics of N mineralization and immobilization. We conclude that D. antarctica and C. quitensis exert a significant influence over C and N cycling in the maritime Antarctic, and that their recent population expansion will have led to significant changes in the amount, type and rate of organic C and N cycling in soil.

Not poles apart: Antarctic soil fungal communities show similarities to those of the distant Arctic

Antarcticas extreme environment and geographical isolation offers a useful platform for testing the relative roles of environmental selection and dispersal barriers influencing fungal communities. The former process should lead to convergence in community composition with other cold environments, such as those in the Arctic. Alternatively, dispersal limitations should minimise similarity between Antarctica and distant northern landmasses. Using high-throughput sequencing, we show that Antarctica shares significantly more fungi with the Arctic, and more fungi display a bipolar distribution, than would be expected in the absence of environmental filtering. In contrast to temperate and tropical regions, there is relatively little endemism, and a strongly bimodal distribution of range sizes. Increasing southerly latitude is associated with lower endemism and communities increasingly dominated by fungi with widespread ranges. These results suggest that micro-organisms with well-developed dispersal capabilities can inhabit opposite poles of the Earth, and dominate extreme environments over specialised local species.

Sebacinales are associates of the leafy liverwort Lophozia excisa in the southern maritime Antarctic.

The leafy liverwort Lophozia excisa, which is colonised by basidiomycete fungi in other biomes and which evidence suggests may be colonised by mycorrhizal fungi in Antarctica, was sampled from Léonie Island in the southern maritime Antarctic (67°36′ S, 68°21′ W). Microscopic examination of plants indicated that fungal hyphae colonised 78% of the rhizoids of the liverwort, apparently by entering the tips of rhizoids prior to growing into their bases, where they formed hyphal coils. Extensive colonisation of stem medullary cells by hyphae was also observed. DNA was extracted from surface-sterilised liverwort tissues and sequenced following nested PCR, using the primer set ITS1F/TW14, followed by a second round of amplification using the ITSSeb3/TW13 primer set. Neighbour-joining analyses showed that the sequences obtained nested in Sebacinales clade B as a 100% supported sister group to Sebacinales sequences from the leafy liverworts Lophozia sudetica, L. incisa and Calypogeia muelleriana sampled from Europe. Direct PCR using the fungal specific primer set ITS1F/ITS4 similarly identified fungi belonging to Sebacinales clade B as the principal colonists of L. excisa tissues. These observations indicate the presence of a second mycothallus in Antarctica and support the previous suggestion that the Sebacinales has a wide geographical distribution.