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.

Two new 12-membered macrolides from the endophytic fungal strain Cladosprium colocasiae A801 of Callistemon viminalis.

Two new polyketide metabolites, the 12-membered macrolides 4-hydroxy-12-methyloxacyclododecane-2,5,6-trione (1) and 12-methyloxacyclododecane-2,5,6-trione (2), were isolated from the endophytic fungal strain Cladosprium colocasiae A801 of the plant Callistemon viminalis, together with five known derivatives. Their structures were fully characterized by means of detailed spectroscopic analysis for new structures, and in comparison with published data for known compounds. The antibacterial, cytotoxic, and α-glucosidase inhibitory activities of the new compounds 1 and 2 were evaluated.

Real-Time PCR Assays to Detect and Distinguish the Rapid 'Ohi'a Death Pathogens Ceratocystis lukuohia and C. huliohia.

Ceratocystis lukuohia and C. huliohia are recently described fungal species that cause rapid 'ohi'a death (ROD) of Metrosideros polymorpha, Hawaii's most abundant and ecologically important native species. Although the pathogens are now widespread on Hawai'i Island, a major effort is underway to study and manage affected forests, and particularly to prevent the disease from spreading to other islands in the State or throughout the Pacific. Rapid and accurate detection is critical. Molecular diagnostic real-time PCR protocols were developed to detect and distinguish the two pathogens, suitable for detection of fungal DNA from extracts of wood, soil, and insect frass. The assays detect as few as 2 to 4 or 16 spores of C. huliohia or C. lukuohia, respectively. These assays are valuable tools for monitoring disease spread and offer a significant advantage over culture-based methods for diagnostics, requiring <1 day to arrive at definitive results.

High hydrostatic pressure processing affects the phenolic profile, preserves sensory attributes and ensures microbial quality of jabuticaba (Myrciaria jaboticaba) juice.

BACKGROUND: Jabuticaba (Myrciaria jaboticaba) is a Brazilian fruit rich in phenolic compounds and much appreciated for its sweet and slightly tangy taste. However, the high perishability of this fruit impairs its economic exploitation, creating an opportunity for the development of innovative products, such as high hydrostatic pressure (HHP) processed juices. We investigated the effect of HHP (200, 350 and 500 MPa for 5, 7.5 and 10 min) on phenolic compounds, antioxidant activity and microbiological quality of jabuticaba juice and the effect of the most effective HHP condition on its sensory acceptance. RESULTS: Pressurization increased total phenolic compound content (up to 38%) and antioxidant activity by FRAP assay (up to 46%), probably by increasing phenolic compound extractability due to tissue damage. Pressurization progressively decreased microbial counts, and colony growth was undetectable at pressures of 350 MPa or 500 MPa. With the exception of aroma, which was 10% lower in pressurized juice at 350 MPa for 7.5 min in relation to unprocessed juice, HHP did not affect sensory acceptance scores. CONCLUSION: Our results show that HHP was effective in ensuring microbiological quality, increasing bioactive potential and maintaining overall acceptance of jabuticaba juice, reinforcing the potential application of this processing technology in bioactive-rich foods. © 2017 Society of Chemical Industry.

Ectomycorrhizal fungal diversity associated with endemic Tristaniopsis spp. (Myrtaceae) in ultramafic and volcano-sedimentary soils in New Caledonia.

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.

ITS phylogeny and taxonomy of Phyllachora species on native Myrtaceae from the Brazilian Cerrado.

Nine Phyllachora species found on hosts belonging to the family Myrtaceae native to the Brazilian Cerrado were described and illustrated. We sequenced nuc rDNA internal transcribed spacer barcode regions for representatives of seven species and conducted phylogenetic analyses, which provided strong support for four new species that we describe as Phyllachora cerradensis, P. ermidensis, P. furnasensis, and P. myrciariae. Catacauma nigerrimum was recombined into P. nigerrima, and a key to the common Phyllachora species on myrtaceous hosts from the Brazilian Cerrado was also included.

Fungal endophyte communities reflect environmental structuring across a Hawaiian landscape.

We surveyed endophytic fungal communities in leaves of a single tree species (Metrosideros polymorpha) across wide environmental gradients (500-5,500 mm of rain/y; 10-22 °C mean annual temperature) spanning short geographic distances on Mauna Loa Volcano, Hawai'i. Using barcoded amplicon pyrosequencing at 13 sites (10 trees/site; 10 leaves/tree), we found very high levels of diversity within sites (a mean of 551 ± 134 taxonomic units per site). However, among-site diversity contributed even more than did within-site diversity to the overall richness of more than 4,200 taxonomic units observed in M. polymorpha, and this among-site variation in endophyte community composition correlated strongly with temperature and rainfall. These results are consistent with suggestions that foliar endophytic fungi are hyperdiverse. They further suggest that microbial diversity may be even greater than has been assumed and that broad-scale environmental controls such as temperature and rainfall can structure eukaryotic microbial diversity. Appropriately constrained study systems across strong environmental gradients present a useful means to understand the environmental factors that structure the diversity of microbial communities.

Saccharopolyspora indica sp. nov., an actinomycete isolated from the rhizosphere of Callistemon citrinus (Curtis).

A novel actinomycete strain, designated VRC122T, was isolated from a Callistemon citrinus rhizosphere sample collected from New Delhi, India, and its taxonomic status was determined by using a polyphasic approach. Strain VRC122T was a Gram-stain-positive, aerobic, non-motile, non-acid-alcohol-fast strain. Phylogenetic analysis based on 16S rRNA gene sequences showed the strain was placed in a well-separated sub-branch within the genus Saccharopolyspora. The highest levels of 16S rRNA gene sequence similarity were found with Saccharopolyspora hirsuta subsp. kobensis JCM 9109T (98.71%), Saccharopolyspora antimicrobica I05-00074T (98.69%) and Saccharopolyspora jiangxiensis W12T (98.66%); 16S rRNA gene sequence similarities with type strains of all other species of the genus Saccharopolyspora were below 98%. Chemosystematic studies revealed that it contained meso-diaminopimelic acid. Arabinose and galactose were the predominant whole-cell sugars. Diagnostic polar lipids were diphosphatidylglycerol, phosphatidylinositol and phosphatidylcholine. MK-9(H6) was the predominant menaquinone. C14:0, C16:0, iso-C15:0, iso-C16:0, iso-C17:0, anteiso-C15:0, anteiso-C17:0, C17:0 cyclo and summed feature 3 (C16:1ω7c and/or C16:1ω6c) were the major cellular fatty acids. The G+C content of the genomic DNA was 69.5 mol%. The results of DNA-DNA hybridization (30%, 22% and 25%, respectively) with type strains of the above-mentioned species, in combination with differences in physiological and biochemical data supported that strain VRC122T represents a novel species of the genus Saccharopolyspora, for which the name Saccharopolyspora indica sp. nov., is proposed. The type strain is VRC122T (=KCTC 29208T=MTCC 11564T=MCC 2206T=ATCC BAA-2551T).

Hannaella pagnoccae sp. nov., a tremellaceous yeast species isolated from plants and soil.

Several independent surveys of yeasts associated with different plant materials and soil led to the proposal of a novel yeast species belonging to the Tremellales clade (Agaricomycotina, Basidiomycota). Analysis of the sequences of the D1/D2 domains and internal transcribed spacer region of the large subunit of the rRNA gene suggested affinity to a phylogenetic lineage that includes Hannaella coprosmaensis, Hannaella oryzae and Hannaella sinensis. Thirty-two isolates were obtained from different sources, including bromeliads, nectar of Heliconia psittacorum (Heliconiaceae), flowers of Pimenta dioica (Myrtaceae), roots and leaves of sugar cane (Saccharum spp.) in Brazil, leaves of Cratoxylum maingayi, Arundinaria pusilla and Vitis vinifera in Thailand, soil samples in Taiwan, and prairie soil in the USA. Sequence analysis of the D1/D2 domains of the large subunit of the rRNA gene showed that the novel species differs from Hannaella coprosmaensis and Hannaella oryzae by 36 and 46 nt substitutions, respectively. A novel species is suggested to accommodate these isolates, for which the name Hannaella pagnoccae sp. nov. is proposed. The type strain is BI118(T) ( = CBS 11142(T) = ATCC MYA-4530(T)).

Australasian sequestrate fungi 18: Solioccasus polychromus gen. & sp. nov., a richly colored, tropical to subtropical, hypogeous fungus.

Solioccasus polychromus gen. & sp. nov., the most brightly colored hypogeous fungus known, is described from Papua New Guinea and tropical northern Australia south into subtropical forests along the Queensland coast and coastal mountains to near Brisbane. Phylogenetic analysis of molecular data places it as a sister genus to Bothia in the Boletineae, a clade of predominantly ectomycorrhizal boletes. Ectomycorrhizal trees, such as members of the Myrtaceae (Eucalyptus, Corymbia, Lophostemon, Melaleuca spp.) and Allocasuarina littoralis, were present usually in mixture or in some cases dominant, so we infer some or all of them to be among the ectomycorrhizal hosts of S. polychromus.

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.

Evaluation of potential responses to invasive non-native species with structured decision making.

In managing invasions and colonizations of non-native species, eradication or control efforts must proceed quickly. There are 2 challenges in taking such quick action. First, managers frequently have to choose among complex and often competing environmental, social, and economic objectives. Second, the effects are highly uncertain. We applied participatory structured decision making (SDM) to develop a response plan for the recent invasion of non-native myrtle rust (Uredo rangelii) in Australia. Structured decision making breaks a complex decision process into 5 steps: identify problems (i.e., decisions to be made), formulate objectives, develop management alternatives, estimate consequences of implementing those alternatives, and select preferred alternatives by evaluating trade-offs among alternatives. To determine the preferred mid- to long-term alternatives to managing the rust, we conducted 2 participatory workshops and 18 interviews with individuals to elicit stakeholders' key concerns and convert them into 5 objectives (minimize management cost, minimize economic cost to industry, minimize effects on natural ecosystems and landscape amenities, and minimize environmental effects associated with use of fungicide) and to identify the 5 management alternatives (full eradication, partial eradication, slow spread, live with it [i.e., major effort invested in mitigation of effects], and do nothing). We also developed decision trees to graphically represent the essence of the decision by displaying the relations between uncertainties and decision points. In the short term or before local expansion of myrtle rust, the do-nothing alternative was not preferred, but an eradication alternative was only recommended if the probability of eradication exceeded about 40%. After the expansion of myrtle rust, the slow-the-spread alternative was preferred regardless of which of the short-term management alternatives was selected at an earlier stage. The participatory SDM approach effectively resulted in informed and transparent response plans that incorporated multiple objectives in decision-making processes under high uncertainty.

Degradation of Staphylococcus aureus Biofilm Using Hydrolytic Enzymes Produced by Amazonian Endophytic Fungi.

Microbial biofilms can cause serious health problems, since, due to their persistent character, they often function as spreaders of contaminants. Hydrolytic enzymes have a number of industrial applications and have been indicated as an alternative to the traditional chemical methods that are used to eradicate microbial biofilms. In this study, we evaluated the ability of enzymatic extracts produced by endophytic fungi isolated from the Amazonian species Myrcia guianensis to remove Staphylococcus aureus biofilms. After culture in liquid medium, the fungal hydrolytic extracts showed amylase (3.77 U/mL), lipase (3.84 U/mL), protease (3.63 U/mL), and xylanase (2.91 U/mL) activity. A 24 h mature S. aureus ATCC6538 biofilm was exposed to each enzyme extract with standardized enzyme activities for 10, 30, and 60 min. The optical density at 630 nm was used to calculate the growth rate (GR%) and the residual biofilm rate (RBR%). The most promising solutions were used in combination, based on a 24 factorial design for 0, 10, 20, and 30 min of exposure. Lipase and protease solutions, when applied separately, were the most effective, and promoted the complete removal of S. aureus biofilms in t10 (lipase) and t30 and t60 (lipase and protease). Of the combined treatments using 1.0 U/mL protease and 0.4 U/mL lipase, total biofilm degradation was observed for all exposure times. Thus, the hydrolases produced by the Amazonian endophytic fungi evaluated here are highlighted as an interesting tool in the fight against microbial biofilms.

New genus of trichomatous coelomycete on Myrcia fenzliana from the Brazilian Cerrado.

Megacoelomyces (type species: Megacoelomyces sanchezii), an ascomycete asexual morph infecting Myrcia fenzliana (Myrtaceae) from the Brazilian Cerrado, is described as a new genus in the Phaeosphaeriaceae (Pleosporales, Dothideomycetes, Ascomycota), based on multilocus phylogeny (three nuclear ribosomal DNA and two protein-coding genes) in addition to morphological (light and scanning electron microscopy) and ecological data.

Specificity between Neotropical tree seedlings and their fungal mutualists leads to plant-soil feedback.

A growing body of evidence obtained largely from temperate grassland studies suggests that feedbacks occurring between plants and their associated soil biota are important to plant community assemblage. However, few studies have examined the importance of soil organisms in driving plant-soil feedbacks in forested systems. In a tropical forest in central Panama, we examined whether interactions between tree seedlings and their associated arbuscular mycorrhizal fungi (AMF) lead to plant-soil feedback. Specifically, do tropical seedlings modify their own AMF communities in a manner that either favors or inhibits the next cohort of conspecific seedlings (i.e., positive or negative feedback, respectively)? Seedlings of two shade-tolerant tree species (Eugenia nesiotica, Virola surinamensis) and two pioneer tree species (Luehea seemannii, Apeiba aspera) were grown in pots containing identical AMF communities composed of equal amounts of inoculum of six co-occurring AMF species. The different AMF-host combinations were all exposed to two light levels. Under low light (2% PAR), only two of the six AMF species sporulated, and we found that host identity did not influence composition of AMF spore communities. However, relative abundances of three of the four AMF species that produced spores were influenced by host identity when grown under high light (20% PAR). Furthermore, spores of one of the AMF species, Glomus geosporum, were common in soils of Luehea and Eugenia but absent in soils of Apeiba and Virola. We then conducted a reciprocal experiment to test whether AMF communities previously modified by Luehea and Apeiba differentially affected the growth of conspecific and heterospecific seedlings. Luehea seedling growth did not differ between soils containing AMF communities modified by Luehea and Apeiba. However, Apeiba seedlings were significantly larger when grown with Apeiba-modified AMF communities, as compared to Apeiba seedlings grown with Luehea-modifed AMF communities. Our experiments suggest that interactions between tropical trees and their associated AMF are species-specific and that these interactions may shape both tree and AMF communities through plant-soil feedback.

Isolation of Cryptococcus gattii molecular type VGIII, from Corymbia ficifolia detritus in Colombia.

An environmental sampling survey was carried out in different areas of Bogotá, Colombia, to obtain isolates of members of the Cryptococcus neoformans/C. gattii species complex from Corymbia ficifolia trees. During a 6-month period in 2007, 128 samples consisting of bark, soil around trunk bases, detritus, seeds and flowers were collected from 91 trees and processed according to standard procedures. The molecular type was determined using URA5 restriction fragment length polymorphism (RFLP) analysis and the mating type was established by PCR using specific primers for Mfalpha and Mfa C. gattii was isolated from 15 of the 128 (11.7%) samples, of which three (20%) were recovered from the red flower extract and the remaining 12 from C. ficifolia detritus. URA5 RFLP analysis revealed that all 15 isolates belonged to the molecular type VGIII and mating type specific PCR revealed that all were mating type a. The isolation of C. gattii from C. ficifolia represents an important finding since this is the first report revealing C. ficifolia as a habitat for C. gattii and adds additional information to the ever growing spectrum of tree species from which C. gattii can be recovered.

Jabuticaba peel extract obtained by microwave hydrodiffusion and gravity extraction: A green strategy to improve the oxidative and sensory stability of beef burgers produced with healthier oils.

Hydrogelled emulsions (HE) from chia and linseed oils (1:1) were made with different concentrations (0, 6, 8, and 10%) of jabuticaba peel extract (JPE) obtained by microwave hydrodiffusion and gravity (MHG) extraction. Burgers (20% fat) were produced with the replacement of 60% of fat by HEs. The oxidative profile and the sensory quality of raw and cooked burgers were evaluated for 120 days (-18 °C). The JPE exhibited 1.72 mg/mL of phenolic compounds and 57,741.67 µmol TE/mL of antioxidant capacity. In addition, the MHG extraction eliminated the mesophilic bacteria from the jabuticaba peel. The burgers made with HE and without the addition of JPE showed a 5-fold increase in TBARS values when compared to the control. On the other hand, the addition of 10% JPE to HE was effective to maintain the lipid oxidation similar to the control until the 60th day of storage. Besides, the incorporation of JPE into HE reduced the sensory defects caused by the lipid reformulation.

Candida metrosideri pro tempore sp. nov. and Candida ohialehuae pro tempore sp. nov., two antifungal-resistant yeasts associated with Metrosideros polymorpha flowers in Hawaii.

Flowers produce an array of nutrient-rich exudates in which microbes can thrive, making them hotspots for microbial abundance and diversity. During a diversity study of yeasts inhabiting the flowers of Metrosideros polymorpha (Myrtaceae) in the Hawai'i Volcanoes National Park (HI, USA), five isolates were found to represent two novel species. Morphological and physiological characterization, and sequence analysis of the small subunit ribosomal RNA (rRNA) genes, the D1/D2 domains of the large subunit rRNA genes, the internal transcribed spacer (ITS) regions, and the genes encoding the largest and second largest subunits of the RNA polymerase II (RPB1 and RPB2, respectively), classified both species in the family Metschnikowiaceae, and we propose the names Candida metrosideri pro tempore sp. nov. (JK22T = CBS 16091 = MUCL 57821) and Candida ohialehuae pro tempore sp. nov. (JK58.2T = CBS 16092 = MUCL 57822) for such new taxa. Both novel Candida species form a well-supported subclade in the Metschnikowiaceae containing species associated with insects, flowers, and a few species of clinical importance. The ascosporic state of the novel species was not observed. The two novel yeast species showed elevated minimum inhibitory concentrations to the antifungal drug amphotericin B (>4 µg/mL). The ecology and phylogenetic relationships of C. metrosideri and C. ohialehuae are also discussed.

Xanalteric acids I and II and related phenolic compounds from an endophytic Alternaria sp. isolated from the mangrove plant Sonneratia alba.

Two new 10-oxo-10H-phenaleno[1,2,3-de]chromene-2-carboxylic acids, xanalteric acids I (1) and II (2), and 11 known secondary metabolites were obtained from extracts of the endophytic fungus Alternaria sp., isolated from the mangrove plant Sonneratia alba collected in China. The metabolites were confirmed to be of fungal origin, and the structures of the new natural products were unambiguously elucidated on the basis of extensive one- and two-dimensional NMR spectroscopic studies and mass spectrometric analysis. The two new compounds 1 and 2 exhibited weak antibiotic activity against multidrug-resistant Staphylococcus aureus. Altenusin (3) displayed broad antimicrobial activity against several additional multidrug-resistant bacterial and fungal strains.

Molecular and phenotypic characterization of three phylogenetic species discovered within the Neofisicoccum parvum/N. ribis complex.

Neofusicoccum parvum and N. ribis are closely related species whose identities often have been confused. These fungal plant pathogens were identified recently as the most abundant species of Botryosphaeriaceae (Ascomycetes) isolated from native Syzygium cordatum trees in South Africa. In another study using multiple gene genealogies from five nuclear loci three undescribed cryptic phylogenetic species as well as N. parvum were identified among 30 of these isolates. The aim of this study was to clarify the identity of the remaining isolates in the N. parvum/N. ribis complex from S. cordatum in South Africa, to describe newly identified cryptic species and to test their pathogenicity. Based on the RNA polymerase II subunit (RPB2) sequence comparisons, the isolates were identified as N. parvum or one of three previously recognized phylogenetic species that are described here as N. cordaticola, N. kwambonambiense and N. umdonicola. These species cannot be separated a priori based on morphological characteristics, although a posteriori analysis of variance showed that the differences in conidial length and width between the species were statistically significant. The isolates of the newly described species as well as N. parvum and N. ribis were tested for pathogenicity on S. cordatum under greenhouse conditions. Isolates representing the three new species were significantly more aggressive than N. parvum and N. ribis with N. kwambonambiense being the most aggressive. This study resolved long-standing questions of identity of species within N. parvum/N. ribis complex and lays a foundation for further studies on this group of pathogens.