Microbial consortium inoculant increases pasture grasses yield in low-phosphorus soil by influencing root morphology, rhizosphere carboxylate exudation and mycorrhizal colonisation.

BACKGROUND: Pasture farming in south-western Australia is challenged by nutrient-poor soils. We assessed the impact of microbial consortium inoculant (MI) and rock mineral fertiliser (MF) on growth, nutrient uptake, root morphology, rhizosphere carboxylate exudation and mycorrhizal colonisation in three pasture grasses - tall fescue (Festuca arundinacea L.), veldt grass (Ehrharta calycina Sm.) and tall wheatgrass (Thinopyrum ponticum L.) grown in low-phosphorus (P) sandy soil in a glasshouse for 30 and 60 days after sowing (DAS). RESULTS: Veldt grass produced the highest specific root length and smallest average root diameter in both growth periods, and had similar shoot weight, root surface area and fine root length (except at 30 DAS) to tall fescue. Compared with the control, MI alone or combined with MF significantly increased shoot and root biomass (except root biomass at 30 DAS), likely due to the significant increases in root surface area and fine root length. Plants supplied with MI + MF had higher shoot N and P contents than those in the MI and the control treatments at 60 DAS. Malate, citrate and trans-aconitate were the major rhizosphere carboxylates exuded at both 30 and 60 DAS. Malate exudation varied among species and treatments in both growth periods, but citrate exudation was consistently higher in the low-P treatments (control and MI) than the MF and MI + MF treatments. CONCLUSION: Microbial consortium inoculant can positively influence pasture production in low-P soil by increasing root surface area and fine root length, whereas exudation of nutrient-mobilising carboxylates (citrate) is dependent more on soil P supply than microbial consortium inoculant. © 2021 Society of Chemical Industry.

Mycotoxins, Phytoestrogens and Other Secondary Metabolites in Austrian Pastures: Occurrences, Contamination Levels and Implications of Geo-Climatic Factors.

Pastures are key feed sources for dairy production and can be contaminated with several secondary metabolites from fungi and plants with toxic or endocrine-disrupting activities, which possess a risk for the health, reproduction and performance of cattle. This exploratory study aimed to determine the co-occurrences and concentrations of a wide range of mycotoxins, phytoestrogens and other secondary metabolites in grazing pastures. Representative samples of pastures were collected from 18 Austrian dairy farms (one sample per farm) between April to October 2019. After sample preparation (drying and milling) the pastures were subjected to multi-metabolite analysis using LC-MS/MS. In total, 68 metabolites were detected, including regulated zearalenone and deoxynivalenol (range: 2.16-138 and 107-505 µg/kg on a dry matter (DM) basis, respectively), modified (3-deoxynivalenol-glucoside, HT-2-glucoside) and emerging Fusarium mycotoxins (e.g., enniatins), ergot alkaloids and Alternaria metabolites along with phytoestrogens and other metabolites. Aflatoxins, fumonisins, T-2 toxin, HT-2 toxin and ochratoxins were not detected. Of the geo-climatic factors and botanical diversity investigated, the environment temperature (average of 2 pre-sampling months and the sampling month) was the most influential factor. The number of fungal metabolites linearly increased with increasing temperatures and temperatures exceeding 15 °C triggered an exponential increment in the concentrations of Fusarium and Alternaria metabolites and ergot alkaloids. In conclusion, even though the levels of regulated mycotoxins detected were below the EU guidance levels, the long-term exposure along with co-occurrence with modified and emerging mycotoxins might be an underestimated risk for grazing and forage-fed livestock. The one-year preliminary data points out a dominant effect of environmental temperature in the diversity and contamination level of fungal metabolites in pastures.

Isolation and Characterization of Cold-Adapted PGPB and Their Effect on Plant Growth Promotion.

Cold-adapted plant growth-promoting bacteria (PGPB) with multiple functions are an important resource for microbial fertilizers with low-temperature application. In this study, culturable cold-adapted PGPB strains with nitrogen fixation and phosphorus solubilization abilities were isolated. They were screened from root and rhizosphere of four dominant grass species in nondegraded alpine grasslands of the Qilian Mountains, China. Their other growth-promoting characteristics, including secretion of indole-3-acetic acid (IAA), production of siderophores and ACC deaminase, and antifungal activity, were further studied by qualitative and quantitative methods. In addition, whether the PGPB strains could still exert plant growth-promoting activity at 4°C was verified. The results showed that 67 isolates could maintain one or more growth-promoting traits at 4°C, and these isolates were defined as cold-adapted PGPB. They were divided into 8 genera by 16S rRNA gene sequencing and phylogenetic analysis, of which Pseudomonas (64.2%) and Serratia (13.4%) were the common dominant genera, and a few specific genera varied among the plant species. A test-tube culture showed that inoculation of Elymus nutans seedlings with cold-adapted PGPB possessing different functional characteristics had a significant growth-promoting effect under controlled low-temperature conditions, including the development of the roots and aboveground parts. Pearson correlation analysis revealed that different growth-promoting characteristics made different contributions to the development of the roots and aboveground parts. These cold-adapted PGPB can be used as excellent strain resources suitable for the near-natural restoration of degraded alpine grasslands or agriculture stock production in cold areas.

Mechanistic understanding of interspecific interaction between a C4 grass and a C3 legume via arbuscular mycorrhizal fungi, as influenced by soil phosphorus availability using a 13 C and 15 N dual-labelled organic patch.

Arbuscular mycorrhizal fungi (AMF) can improve plant nutrient acquisition, either by directly supplying nutrients to plants or by promoting soil organic matter mineralization, thereby affecting interspecific plant relationships in natural communities. We examined the mechanism by which the addition of P affects interspecific interactions between a C4 grass (Bothriochloa ischaemum, a dominant species in natural grasslands) and a C3 legume (Lespedeza davurica, a subordinate species in natural grasslands) via AMF and plant growth, by continuous 13 C and 15 N labelling, combined with soil enzyme analyses. The results of 15 N labelling revealed that P addition affected the shoot uptake of N via AMF by B. ischaemum and L. davurica differently. Specifically, the addition of P significantly increased the shoot uptake of N via AMF by B. ischaemum but significantly decreased that by L. davurica. Interspecific plant interactions via AMF significantly facilitated the plant N uptake via AMF by B. ischaemum but significantly inhibited that by L. davurica under P-limited soil conditions, whereas the opposite effect was observed in the case of excess P. This was consistent with the impact of interspecific plant interaction via AMF on arbuscular mycorrhizal (AM) benefit for plant growth. Our data indicate that the capability of plant N uptake via AMF is an important mechanism that influences interspecific relationships between C4 grasses and C3 legumes. Moreover, the effect of AMF on the activities of the soil enzymes responsible for N and P mineralization substantially contributed to the consequence of interspecific plant interaction via AMF for plant growth.

Antibacterial activity of bacteria isolated from Phragmites australis against multidrug-resistant human pathogens.

Background: Rising number of multidrug-resistant human pathogens demands novel antibiotics: to this aim, unexplored natural sources are investigated to find new compounds. In this context, bacteria associated to medicinal plants, including Phragmites australis, might represent an important source of antimicrobial compounds. Materials & methods: In the present work, 21 bacterial endophytes isolated from P. australis roots were tested, by cross-streaking, for their inhibitory activity against 36 multidrug-resistant pathogens isolated from food, clinical patients and hospitals. Results & conclusion: Seven endophytes, belonging to Pseudomonas and Stenotrophomonas, were able to inhibit the growth of most of the target strains. In conclusion, this preliminary work could pave the way for the discovery of new antibiotics against superbugs.

Lay abstract In the present work, 21 bacteria associated to Phragmites australis roots were tested for their inhibitory activity against 36 human pathogens isolated from food, clinical patients and/or hospitals, which have the ability to escape several commonly used antibiotics. Seven out of 21 bacteria associated to P. australis were able to inhibit the growth of most of target pathogens. This preliminary work could pave the way to the discovery of new antimicrobial compounds active against bacterial pathogens that cannot be killed using several antibiotics.

Microbial community dynamics in phyto-thermotherapy baths viewed through next generation sequencing and metabolomics approach.

Phyto-thermotherapy is a treatment consisting in immersing oneself in baths of self-heating alpine grass, to benefit of the heat and rich aromatic components released by the process. The aim of this study was to characterize the bacterial and fungal diversity of three phyto-thermal baths (PTB) performed in three different months, and to compare the data with the profile of the volatile organic compounds (VOCs) of the process. All the data collected showed that PTBs were structured in two stages: the first three days were characterised by an exponential rise of the temperature, a fast bacterial development, higher microbial diversity and higher concentrations of plant aliphatic hydrocarbons. The second stage was characterised by a stable high temperature, shrinkage of the microbial diversity with a predominance of few bacterial and fungi species and higher concentrations of volatiles of microbial origin. Erwinia was the dominant microbial species during the first stage and probably responsible of the self-heating process. In conclusion, PTBs has shown both similarities with common self-heating processes and important peculiarities such as the absence of pathogenic bacteria and the dominance of plant terpenoids with health characteristics among the VOCs confirming the evidence of beneficial effects in particular in the first three days.

Influence of dragon bamboo with different planting patterns on microbial community and physicochemical property of soil on sunny and shady slopes.

Dragon bamboo (Dendrocalamus giganteus) is a giant sympodial bamboo species widely distributed in Asia. However, it remains unclear how dragon bamboo and soil microbes interact to affect soil properties. In this study, we investigated the planting patterns (semi-natural and artificial) on different slopes (sunny and shady) to determine the effects on soil properties and microbial community. The results showed that the soil in which dragon bamboo was grown was acidic, with a pH value of ∼5. Also, the soil organic matter content, nitrogen hydrolysate concentration, total nitrogen, available potassium, and total potassium of the dragon bamboo semi-natural forest significantly improved, especially on the sunny slope. In contrast, the available phosphorus level was higher in the artificial bamboo forest, probably owing to the phosphate fertilizer application. The bacterial and fungal diversity and the bacterial abundance were all higher on the sunny slope of the semi-natural forest than those in the other samples. The microbial operational taxonomic units (OTUs) shared between the shady and sunny slopes accounted for 47.8-62.2%, but the core OTUs of all samples were only 24.4-30.4% of each sample, suggesting that the slope type had a significant effect on the microbial community. Some acidophilic microbes, such as Acidobacteria groups, Streptomyces and Mortierella, became dominant in dragon bamboo forest soil. A PICRUSt analysis of the bacterial functional groups revealed that post-translational modification, cell division, and coenzyme transport and metabolism were abundant in the semi-natural forest. However, some microorganisms with strong stress resistance might be activated in the artificial forest. Taken together, these results illustrated the influence of dragon bamboo growth on soil physicochemical property and microbial community, which might help understand the growth status of dragon bamboo under different planting patterns.

The effect of an additive containing three Lactobacillus species on the fermentation pattern and microbiological status of silage.

BACKGROUND: Appropriate combinations of lactic acid bacteria (LAB) strains should be selected to optimize the ensiling process, and the additives should be adjusted to the ensiled forage crops. The aim of this study was to determine the effect of inoculation with three Lactobacillus species on the chemical parameters and microbiological quality (beneficial and harmful microbiota) of grass silage. RESULTS: Three species: L. paracasei (LPa), L. brevis (LB) and L. plantarum (LPl), isolated from sugar beet silage and characterized based on 16S rDNA sequences and biochemical parameters, were analyzed in the study. Single strains and their combinations were used as silage inoculants. The basic chemical and microbiological (qPCR) parameters of silages were determined. Based on the results of agglomerative hierarchical clustering (AHC) and principal component analysis (PCA) it was determined that silages inoculated with single LAB strains and LPa + LB and commercial additive (0+) were the best quality ones, particularly with regard to microbiological parameters and they effectively lowered the pH value. A consortium of three Lactobacillus species had no influence on silage quality, whereas LPa + LB and LPl + LB combinations as well as a commercial additive exerted positive effects. Inoculation inhibited the growth of toxin-producing fungi. CONCLUSION: Only the appropriate LAB composition can improve the quality of the ensiled material (antagonistic relationship). Only the LPa + LB combination was able to improve the value of low dry silage; nevertheless, almost all combinations were able to reduced concentrations of toxin-producing fungi. © 2019 Society of Chemical Industry.

Arbuscular mycorrhizal fungi alleviate phosphorus limitation by reducing plant N:P ratios under warming and nitrogen addition in a temperate meadow ecosystem.

Global change apart from ecosystem processes also influences the community structure of key organisms, such as arbuscular mycorrhizal fungi (AMF). We conducted a 3-year experiment where we suppressed with benomyl mycorrhiza to understand how AMF alter the plant community structure under warming and nitrogen (N) addition. The elemental content and foliar tissue stoichiometry of the dominant species Leymus chinensis and the subordinate species Puccinellia tenuiflora were studied along with soil nutrient stoichiometries. Overall, N addition enhanced plant N: phosphorus (P) ratios at a greater level than experimental warming did. Under global change conditions, AMF symbionts significantly increased soil available P concentrations, promoted plant P absorption and decreased the plant N:P ratios. AMF alleviate P limitation by reducing plant N:P ratios. Our results highlight that the negative influence of global change on plant productivity might cancel each other out through the additive effects of AMF and that global change will increase the dependency of plants on their mycorrhizal symbionts.

Arbuscular Mycorrhizal Fungi Alter Plant and Soil C:N:P Stoichiometries Under Warming and Nitrogen Input in a Semiarid Meadow of China.

Ecological stoichiometry has been widely used to determine how plant-soil systems respond to global change and to reveal which factors limit plant growth. Arbuscular mycorrhizal fungi (AMF) can increase plants' uptake of nutrients such as nitrogen (N) and phosphorus (P), thereby altering plant and soil stoichiometries. To understand the regulatory effect of AMF feedback on plants and soil stoichiometry under global change, a microcosm experiment was conducted with warming and N input. The C4 grass Setaria viridis, C3 grass Leymus chinensis, and Chenopodiaceae species Suaeda corniculata were studied. The results showed that the mycorrhizal benefits for the C4 grass S. viridis were greater than those for the C3 grass L. chinensis, whereas for the Chenopodiaceae species S. corniculata, AMF symbiosis was antagonistic. Under N input and a combination of warming and N input, AMF significantly decreased the N:P ratios of all three species. Under N input, the soil N content and the N:P ratio were decreased significantly in the presence of AMF, whereas the soil C:N ratio was increased. These results showed that AMF can reduce the P limitation caused by N input and improve the efficiency of nutrient utilization, slow the negative influence of global change on plant growth, and promote grassland sustainability.

Arbuscular mycorrhizal fungi alleviate boron toxicity in Puccinellia tenuiflora under the combined stresses of salt and drought.

To investigate the effect of arbuscular mycorrhizal fungi (AMF) on boron (B) toxicity in plants under the combined stresses of salt and drought, Puccinellia tenuiflora was grown in the soil with the inoculation of Funneliformis mosseae and Claroideoglomus etunicatum. After three weeks of treatment, the plants were harvested to determine mycorrhizal colonization rates, plant biomass, as well as tissue B, phosphorus, sodium, and potassium concentrations. The results show that the combined stresses reduced mycorrhizal colonization. Mycorrhizal inoculation significantly increased plant biomass while reduced shoot B concentrations. Mycorrhizal inoculation also slightly increased shoot phosphorus and potassium concentrations, and reduced shoot sodium concentrations. F. mosseae and C. etunicatum were able to alleviate the combined stresses of B, salt, and drought. The two fungal species and their combination showed no significant difference in the alleviation of B toxicity. It is inferred that AMF is able to alleviate B toxicity in P. tenuiflora by increasing biomass and reducing tissue B concentrations. The increase in plant phosphorus and potassium, as well as the decrease in sodium accumulation that induced by AMF, can help plant tolerate the combined stresses of salt and drought. Our findings suggest that F. mosseae and C. etunicatum are potential candidates for facilitating the phytoremediation of B-contaminated soils with salt and drought stress.

Decay of the water reed Phragmites communis caused by the white-rot fungus Phlebia tremellosa and the influence of some environmental factors.

The strain Phlebia tremellosa SBUG 1630 isolated from a thatched roof in Northern Germany is capable of colonizing and degrading effectively the water reed Phragmites communis. Within 96 h after inoculation, mycelia covered both the outer and the inner surface of reed shoot fragments as observed by scanning electron microscopy. Interestingly, top culm sections and culm edges were particularly susceptible towards fungal degradation. The weight loss of culms reached 20-73% depending on the environmental conditions applied during the incubation of 70 days. Reed degradation was stable at pH 4 to pH 8 and optimal between 25 and 30 °C. Short-term incubation at elevated temperatures (37 to 55 °C) affected the fungal reed degradation to only a minor extent, whereas > 18 h at 55 °C completely inhibited fungal growth and reed degradation. Supplementation with 43 mM NH4Cl enhanced the reed degradation up to 9%. In contrast, the addition of diammonium tartrate increased the weight loss of the samples considerably up to 16% at 344 mM. Furthermore, reed degradation by P. tremellosa was increased by supplementing the test medium with Mn (99 to 1584 µM), Cu (150 to 300 µM), and less significantly phosphate (4 mM), Zn (37 to 74 µM), and Ag (76 µM) after 70 days. In addition, activities of the ligninolytic enzymes laccase (max. 27.4 nmol ml-1 min-1) and lignin peroxidase (max. 22.8 nmol ml-1 min-1) were rather low in nitrogen-limited medium, whereas considerably higher levels of manganese peroxidase (max. 635.9 nmol ml-1 min-1) were observed.

Isolation and molecular identification of lactic acid bacteria from King grass and their application to improve the fermentation quality of sweet Sorghum.

The aim of the present study was isolation and molecular identification of lactic acid bacteria from King grass and their application to improve the fermentation quality of sweet Sorghum. Seventy-six strains of LAB were isolated; five strains were selected for Physiological and morphological tests and 16S rRNA sequencing. All five strains grew at different pH 3.5-8.0, different temperature 35, 40, 45, 50 °C and different NaCl concentrations 3, 6.5, 9.5%. Strains HDASK were identified Lactobacillus plantarum and SK3907, SK2A32, SK3A42 and ASKDD Pediococcus acidilactici. Three isolated strains and one commercial strain were added to sweet sorghum. Silage was prepared of four treatments and one control with three replicates as control (SKC, adding 2 ml/kg sterilizing water), L. plantarum commercial bacteria (SKP), L. plantarum (HDASK) isolated from King grass (SKA), P. acidilactici (SK3907) isolated from King grass (SKB) and P. acidilactici (ASKDD) isolated from King grass (SKD). All silage were prepared using polyethylene terephthalate bottles, and incubated at room temperature for different ensiling days. The level of pH, acetic acid, NH3-N, water soluble carbohydrate and butyric acid was significantly (P < 0.05) decreased. Lactic acid, ethanol and propionic acid (PA) was significantly (P < 0.05) increased in treatments compared to control. The dry matter, propionic acid neutral detergent fiber, acid detergent fiber did not significantly (P < 0.05) differ among the treatments but the values were increased and decreased. The number of yeast, mold and LAB were significantly (P < 0.05). It is suggested that the supplementation of LAB could enhanced the fermentation quality of sweet Sorghum silage.

Cytogenetic analysis and mapping of leaf rust resistance in Aegilops speltoides Tausch derived bread wheat line Selection2427 carrying putative gametocidal gene(s).

Leaf rust (Puccinia triticina) is a major biotic stress affecting wheat yields worldwide. Host-plant resistance is the best method for controlling leaf rust. Aegilops speltoides is a good source of resistance against wheat rusts. To date, five Lr genes, Lr28, Lr35, Lr36, Lr47, and Lr51, have been transferred from Ae. speltoides to bread wheat. In Selection2427, a bread wheat introgresed line with Ae. speltoides as the donor parent, a dominant gene for leaf rust resistance was mapped to the long arm of chromosome 3B (LrS2427). None of the Lr genes introgressed from Ae. speltoides have been mapped to chromosome 3B. Since none of the designated seedling leaf rust resistance genes have been located on chromosome 3B, LrS2427 seems to be a novel gene. Selection2427 showed a unique property typical of gametocidal genes, that when crossed to other bread wheat cultivars, the F1 showed partial pollen sterility and poor seed setting, whilst Selection2427 showed reasonable male and female fertility. Accidental co-transfer of gametocidal genes with LrS2427 may have occurred in Selection2427. Though LrS2427 did not show any segregation distortion and assorted independently of putative gametocidal gene(s), its utilization will be difficult due to the selfish behavior of gametocidal genes.

Microbes participated in macrophyte leaf litters decomposition in freshwater habitat.

Knowledge of aquatic microbes involved in macrophyte leaf litter decomposition is still scarce in freshwater lakes. In situ experiments (150 days) were conducted to study the decomposition processes of macrophyte leaf litters: Zizania latifolia (Zl), Hydrilla verticillata (Hv) and Nymphoides peltata (Np). The decomposition of Np leaf litter was fastest, whereas Zl was slowest. The alpha diversity of both bacterial and fungal communities significantly increased, and their community structures showed significant variations over time. For bacteria, the relative abundance of Gammaproteobacteria decreased, whereas that of Firmicutes, Betaproteobacteria, Deltaproteobacteria and Alphaproteobacteria increased. The dominant fungal phylum Cryptomycota increased significantly in all of the three macrophytes. Both bacteria and fungi were significantly correlated with the dynamics of total phosphorous in the water and the carbon content of the leaf litters. The dynamics of nitrogen content, phosphorous content and N/P ratio of the leaf litters have more influences on fungal communities than on bacteria. In addition, cellulase and xylanase activities were significantly correlated with bacterial and fungal communities, respectively, thereby reflecting the niches differentiation and cooperation between bacteria and fungi on litter decomposition. This work contributes to the understanding of microbially involved carbon and nutrient cycling in macrophyte-dominated freshwater ecosystems.

Influence of microbial community diversity and function on pollutant removal in ecological wastewater treatment.

Traditional wastewater treatments based on activated sludge often encounter the problems of bulking and foaming, as well as malodor. To solve these problems, new treatment technologies have emerged in recent decades, including the ecological wastewater treatment process, which introduces selected local plants into the treatment system. With a focus on the underlying mechanisms of the ecological treatment process, we explored the microbial community biomass, composition, and function in the treatment system to understand the microbial growth in this system and its role in pollutant removal. Flow cytometry analysis revealed that ecological treatment significantly decreased influent bacterial quantity, with around 80% removal. 16S rRNA gene sequencing showed that the ecological treatment also altered the bacterial community structure of the wastewater, leading to a significant change in Comamonadaceae in the effluent. In the internal ecological system, because most of microbes aggregate in the plant rhizosphere and the sludge under plant roots, we selected two plant species (Nerium oleander and Arundo donax) to study the characteristics of rhizosphere and sludge microbes. Metagenomic results showed that the microbial community composition and function differed between the two species, and the microbial communities of A. donax were more sensitive to seasonal effects. Combined with their greater biomass and abundance of metabolic genes, microbes associated with N. oleander showed a greater contribution to pollutant removal. Further, the biodegradation pathways of some micropollutants, e.g., atrazine, were estimated.

Elevated CO2 and nitrogen addition have minimal influence on the rhizospheric effects of Bothriochloa ischaemum.

The influence of elevated CO2 and nitrogen (N) addition on soil microbial communities and the rhizospheric effects of Bothriochloa ischaemum were investigated. A pot-cultivation experiment was conducted in climate-controlled chambers under two levels of CO2 (400 and 800 µmol mol-1) and three levels of N addition (0, 2.5, and 5 g N m-2 y-1). Soil samples (rhizospheric and bulk soil) were collected for the assessment of soil organic carbon (SOC), total N (TN), total phosphorus (TP), basal respiration (BR), and phospholipid fatty acids (PLFAs) 106 days after treatments were conducted. Elevated CO2 significantly increased total and fungal PLFAs in the rhizosphere when combined with N addition, and N addition significantly increased BR in the rhizosphere and total, bacterial, fungal, Gram-positive (G+), and Gram-negative (G-) PLFAs in both rhizospheric and bulk soil. BR and total, bacterial, G+, and G+/G- PLFAs were significantly higher in rhizospheric than bulk soil, but neither elevated CO2 nor N addition affected the positive rhizospheric effects on bacterial, G+, or G+/G- PLFAs. N addition had a greater effect on soil microbial communities than elevated CO2, and elevated CO2 and N addition had minor contributions to the changes in the magnitude of the rhizospheric effects in B. ischaemum.

Contrasting bacterial communities in two indigenous Chionochloa (Poaceae) grassland soils in New Zealand.

The cultivation of grasslands can modify both bacterial community structure and impact on nutrient cycling as well as the productivity and diversity of plant communities. In this study, two pristine New Zealand grassland sites dominated by indigenous tall tussocks (Chionochloa pallens or C. teretifolia) were examined to investigate the extent and predictability of variation of the bacterial community. The contribution of free-living bacteria to biological nitrogen fixation is predicted to be ecologically significant in these soils; therefore, the diazotrophic community was also examined. The C. teretifolia site had N-poor and poorly-drained peaty soils, and the C. pallens had N-rich and well-drained fertile soils. These soils also differ in the proportion of organic carbon (C), Olsen phosphorus (P) and soil pH. The nutrient-rich soils showed increased relative abundances of some copiotrophic bacterial taxa (including members of the Proteobacteria, Bacteroidetes and Firmicutes phyla). Other copiotrophs, Actinobacteria and the oliogotrophic Acidobacteria showed increased relative abundance in nutrient-poor soils. Greater diversity based on 16S rRNA gene sequences and the Tax4Fun prediction of enhanced spore formation associated with nutrient-rich soils could indicate increased resilience of the bacterial community. The two sites had distinct diazotrophic communities with higher diversity in C. teretifolia soils that had less available nitrate and ammonium, potentially indicating increased resilience of the diazotroph community at this site. The C. teretifolia soils had more 16S rRNA gene and nifH copies per g soil than the nutrient rich site. However, the proportion of the bacterial community that was diazotrophic was similar in the two soils. We suggest that edaphic and vegetation factors are contributing to major differences in the composition and diversity of total bacterial and diazotrophic communities at these sites. We predict the differences in the communities at the two sites will result in different responses to environmental change.

An empirical investigation of the possibility of adaptability of arbuscular mycorrhizal fungi to new hosts.

Little is known about the adaptive capacity of arbuscular mycorrhizal (AM) fungi to novel hosts. Here we assessed the possibility of two heterospecific AM fungal isolates to adaptively change, in terms of host biomass response, as a function of host plant identity, over the course of a growing season. First, we produced pure inocula of Rhizophagus clarus and Rhizophagus intraradices, each starting from a single spore. Second, we "trained" each isolate individually in a community with two plants, sudangrass (Sorgum bicolour subsp. drummondii) and leek (Aliium ampeloprasum var. porrum), using a dual-compartment system to allow the establishment of a common mycorrhizal network between the two hosts. Third, we conducted a greenhouse experiment to reciprocally test each "trained" clone, obtained from each compartment, either with the same (home), or the other host (away) under two contrasting phosphorus levels. Overall, results did not support adaptive responses of the AM fungi to their hosts (i.e., greater host biomass under "home" relative to "away" conditions), but the opposite (i.e., greater host biomass under "away" relative to "home" conditions) was more frequently observed. These changes in AM fungal symbiotic functioning open the possibility for relatively rapid genetic change of arbuscular mycorrhizal fungi in response to new hosts, which represents one step forward from in vitro experiments.

Uso de Leersia hexandra (Poaceae) en la fitorremediación de suelos contaminados con petróleo fresco e intemperizado / Use of Leersia hexandra (Poaceae) for soil phytoremediation in soils contaminated with fresh and weathered oil

ResumenLa industria petrolera ha generado derrames crónicos de petróleo y su acumulación en Gleysoles en zonas anegadas en el estado de Tabasco, en el sureste de México. El anegamiento es un factor que limita el uso de tecnologías de remediación por el alto costo y los bajos niveles de degradación del petróleo, sin embargo, Leersia hexandra Sw. es un pasto que crece en estas zonas contaminadas con petróleo intemperizado. El objetivo del estudio fue evaluar la densidad de bacterias, producción de biomasa vegetal y fitorremediación de L.hexandra en suelo contaminado con petróleos fresco e intemperizado, bajo condiciones experimentales de anegamiento. Se realizaron dos experimentos (E1 y E2) en un túnel de plástico. El E1 se basó en ocho dosis: 6 000, 10 000, 30 000, 60 000, 90 000, 120 000, 150 000 y 180 000 mg kg-1 base seca (b.s.) de hidrocarburos totales de petróleo fresco (HTPF), y en el E2 se evaluaron cinco dosis: 14 173, 28 400, 50 598, 75 492 y 112 142 mg kg-1 b. s. de hidrocarburos totales de petróleo intemperizado (HTPI), con ocho repeticiones en cada experimento, además se utilizó un testigo con 2 607 mg kg-1 b. s. de HTP de origen biogénico. Las variables evaluadas a los tres y seis meses fueron 1) densidad microbiana de las bacterias fijadoras de nitrógeno de vida libre totales (BFN), del grupo Azospirillum (AZP) y Azotobacter (AZT), por cuenta viable en placa seriada; 2) producción de materia seca total (MS), se cuantificó por el peso seco por gravimetría, y 3) el porcentaje de descontaminación de los hidrocarburos (DSC) por extracción en equipo soxhlet. En suelos con HTPF, la población de BFN, AZP y AZT se estimuló hasta cinco veces más que el tratamiento testigo a los tres y seis meses; sin embargo, concentraciones de 150 000 y 180 000 mg kg-1 b. s. inhibieron entre un 70 y 89 % la densidad bacteriana. A su vez, en suelos con PI, la inhibición se registró hasta en un 90 %, a excepción del tratamiento con 14 173 mg kg-1 b. s., el cual estimuló las BNF y AZT en 2 y 0.10 veces más que testigo, respectivamente. La producción de MS fue continua en los experimentos hasta los seis meses, con valores de 63 y 89 g en PF y PI, respectivamente; sin diferencias significativas con el testigo (p ≤ 0.05). El DSC alcanzó valores del 66 % al 87 % en HTPF como HTPI a los seis meses, respectivamente. Estos resultados demuestran la habilidad del L. hexandra para desarrollar una rizósfera con alta densidad de BFN, producir biomasa vegetal y fitorremediar Gleysoles con petróleo fresco e intemperizado en ambientes tropicales inundados.

Abstract:The oil industry has generated chronic oil spills and their accumulation in wetlands of the state of Tabasco, in Southeastern Mexico. Waterlogging is a factor that limits the use of remediation technologies because of its high cost and low levels of oil degradation. However, Leersia hexandra is a grass that grows in these contaminated areas with weathered oil. The aim of the study was to evaluate the bacteria density, plant biomass production and phytoremediation of L. hexandra in contaminated soil. For this, two experiments in plastic tunnel were performed with fresh (E1) and weathered petroleum (E2) under waterlogging experimental conditions. The E1 was based on eight doses: 6 000, 10 000, 30 000, 60 000, 90 000, 120 000, 150 000 and 180 000 mg.kg-1 dry basis (d. b.) of total petroleum hydrocarbons fresh (TPH-F), and the E2, that evaluated five doses: 14 173, 28 400, 50 598, 75 492 and 112 142 mg. kg-1 d. b. of total petroleum hydrocarbons weathered (TPH-W); a control treatment with 2 607 mg.kg-1 d. b. was used. Each experiment, with eight replicates per treatment, evaluated after three and six months: a) microbial density of total free-living nitrogen-fixing bacteria (NFB) of Azospirillum (AZP) and Azotobacter group (AZT), for viable count in serial plate; b) dry matter production (DMP), quantified gravimetrically as dry weight of L. hexandra; and c) the decontamination percentage of hydrocarbons (PDH) by Soxhlet extraction. In soil with TPH-F, the NFB, AZP y AZT populations were stimulated five times more than the control both at the three and six months; however, concentrations of 150 000 and 180 000 mg.kg-1 d. b. inhibited the bacterial density between 70 and 89 %. Likewise, in soil with TPH-W, the FNB, AZP and AZT inhibitions were 90 %, with the exception of the 14 173 mg.kg-1 d. b. treatment, which stimulated the NFB and AZT in 2 and 0.10 times more than the control, respectively. The DMP was continued at the six months in the experiments, with values of 63 and 89 g in fresh and weathered petroleum, respectively; had no significant differences with the control (p≤0.05). The PDH reached values of 66 to 87 % both TPH-F and TPH-W at six months, respectively. These results demonstrated the ability the L. hexandra rhizosphere to stimulate the high NFB density, vegetal biomass production and phytoremediation of contaminated soils (with fresh and weathered petroleum), in a tropical waterlogging environment. Rev. Biol. Trop. 65 (1): 21-30. Epub 2017 March 01.