Compost application boosts soil restoration in highly disturbed hillslope vineyard.

A field trial was carried out to investigate the effects of compost application on a young Cabernet sauvignon vineyard located in a hilly area in the North-East of Italy and subjected to land terracing before plantation. The use of a compost based on manure, pruning residues and pomace at a rate of 65 t ha-1 was compared to the mineral fertilization regime recommended for the vineyards in the area (NPK: 80, 50, 200 kg ha-1). A multi-factorial approach that considered soil chemical properties, microbial community structure and function, vine nutritional and vegetative indexes, yield and quality parameters was applied in the attempt of depict interrelated effects of compost on all these factors. Results of this study show that the application of compost for three consequent years greatly increased soil organic matter content and improved the mineral nutrient availability in the soil. Soil biological fertility showed a slow but significant response to compost addition as from the second year of treatment microbial growth and enzyme activity were increased compared to those of the inorganic fertilization, with special regard to enzymes involved in P cycle. A shift in the soil microbial community structure was also observed in compost-treated soil, with higher presence of copiotrophic bacteria, indicators of soil quality, and phosphorus solubilizing bacteria. A decrease of pathogenic fungal strains was also observed. Organic fertilization increased plant nutrient uptake and vegetative growth compared to those observed in chemically fertilized vines. A trend toward increased yield and improvements for some grape quality parameters such as acidity and pH were observed in the first year of production. These results provide evidence that compost can boost soil fertility restoration in vineyard disturbed by land terracing, allowing for agronomic performances comparable or even improved than those of chemically fertilized vines.

Perenniality, more than genotypes, shapes biological and chemical rhizosphere composition of perennial wheat lines.

Perennial grains provide various ecosystem services compared to the annual counterparts thanks to their extensive root system and permanent soil cover. However, little is known about the evolution and diversification of perennial grains rhizosphere and its ecological functions over time. In this study, a suite of -OMICSs - metagenomics, enzymomics, metabolomics and lipidomics - was used to compare the rhizosphere environment of four perennial wheat lines at the first and fourth year of growth in comparison with an annual durum wheat cultivar and the parental species Thinopyrum intermedium. We hypothesized that wheat perenniality has a greater role in shaping the rhizobiome composition, biomass, diversity, and activity than plant genotypes because perenniality affects the quality and quantity of C input - mainly root exudates - hence modulating the plant-microbes crosstalk. In support of this hypothesis, the continuous supply of sugars in the rhizosphere along the years created a favorable environment for microbial growth which is reflected in a higher microbial biomass and enzymatic activity. Moreover, modification in the rhizosphere metabolome and lipidome over the years led to changes in the microbial community composition favoring the coexistence of more diverse microbial taxa, increasing plant tolerance to biotic and abiotic stresses. Despite the dominance of the perenniality effect, our data underlined that the OK72 line rhizobiome distinguished from the others by the increase in abundance of Pseudomonas spp., most of which are known as potential beneficial microorganisms, identifying this line as a suitable candidate for the study and selection of new perennial wheat lines.

Taxonomic and functional responses of soil and root bacterial communities associated with poplar exposed to a contamination gradient of phenanthrene.

Polycyclic aromatic hydrocarbon (PAH) contamination of industrial wasteland soils affects microbial diversity, but little is known about the dose-response effects of such contaminants on taxonomic and functional diversities of rhizospheric and plant endophytic bacteria. This study focused on the response of soil and root bacterial communities associated to poplar grown in a contamination gradient of phenanthrene (PHE). It was hypothesized that the increase in contamination would modify gradually the bacterial diversity and functions. The effects of the PHE contamination were limited to soil communities and did not affect the poplar root endophytome where Streptomyces and Cutibacterium were the most abundant genera. Along the PHE gradient, alpha-diversity indices decreased and the community structure of soil bacteria at the taxonomic level shifted. The abundance of genes involved in PAH-degradation pathways and the relative proportion of certain microbial taxa such as Polaromonas, Sphingopyxis, Peredibacter, Phenylobacterium, Ramlibacter, Sphingomonas, and Pseudomonas, often described as potential PAH biodegraders, increased with the PHE concentration in the soil community. Conversely, the contamination negatively impacted other taxa like Nocardioides, Streptomyces, Gaiella, Solirubrobacter, Bradyrhizobium, and Nitrospira. Functional inference and enzymatic activity measurements revealed that some bacterial functions related to carbon, nitrogen and phosphorus cycles were modified in soil throughout the PHE gradient. This study allowed a deeper understanding of the complex plant-bacteria interactions in the case of soil PAH contamination and the potential impact on soil functioning.

Influence of earthworms on the microbial properties and extracellular enzyme activities during vermicomposting of raw and distilled grape marc.

The treatment of winery wastes by using appropriate management technologies is of utmost need in order to reduce to a minimum their disposal and avoid negative environmental impacts. This is of particular interest for grape marc, the main solid by-product of the winery industry. However, comparative studies on a pilot-scale dealing with the impact of earthworms on marc derived from both red and white grape varieties during vermicomposting are still scarce. The present study sought to evaluate the changes in the biochemical and microbiological properties of red and white raw marc in the presence and the absence of the earthworm species Eisenia andrei. The distilled marc obtained through distillation of the red grape marc was also considered under this scenario. Samples were taken after 14, 28, 42, and 63 days of vermicomposting. On day 14 earthworms led to a pronounced increase in most of the enzymatic activities, but only in those vermireactors fed with raw marc from the red grape variety. Alfa- and beta-glucosidase as well as chitinase and leucine-aminopeptidase activities were between 3 to 5-times higher relative to the control, while alkaline phosphomonoesterase was even up to 14-fold higher with earthworm presence. From day 28 onwards the magnitude of earthworms' effect on the studied enzymes was also dependent on the type of grape marc. Reduced values of basal respiration, ranging between 200 and 350 mg CO2 kg OM h-1 and indicative of stabilized materials were found in the resulting vermicomposts. Moreover, the content of macro- and micronutrients in the end products matched with those considered to have the quality criteria of a good vermicompost. Altogether, these findings reinforce the effectiveness of vermicomposting for the biological stabilization of grape marc with the dual purpose of fertilizer production and environmental protection.

Spent coffee grounds by-products and their influence on soil C-N dynamics.

SCG are a bio-waste generated in great amount worldwide which are attractive as soil amendment for their high content of organic matter and nutritive elements. Nevertheless, several studies have shown that soil application of untreated SCG has detrimental agronomic and environmental effects due to their high degradability and content of noxious compounds (phenols, caffeine, and tannins). However, SCG can be valorised, in the frame of circular economy, by extraction of energy and valuable products (carbohydrates, proteins, bio-oil, bio-diesel) and generation of solid by products (biochar, hydrochar, compost) that can be utilized as soil fertilizers and amendments. Therefore, the aim of this work was the characterization of different solid SCG by-products (as second-generation products) and their assessment as effective organic amendments. The novelty of this study is that for the first time 8 different by-products derived from the same SCG were characterized and comparatively evaluated for their impact on the C and N cycles of soil. SCG was collected and treated to generate 8 different SCG by-products (biochars produced at 270 and 400 °C, hydrochars produced at 160 and 200 °C, vermicompost, defatted SCG and biochars produced from defatted SCG at 270 and 400 °C). SCG and derived by-products were characterized for SEM micromorphology, pH and EC values, and C, N, H, O, volatile matter, fixed C, LOI, carbonates, water soluble C and N, NO3- and NH4+ content. SCG and SCG by-products assessment as organic amendments was performed with an incubation experiment. The residues were added (2.5%) to a moist Mediterranean agricultural soil and the amended soil samples were placed in mesocosms and incubated at 20 °C for 30 days. During incubation, CO2 and N2O emissions were measured every 6 h by means of a gas chromatography automated system for GHG sampling and measurement. The percentage of added C remaining (CR) in the soil was calculated by fitting the cumulative respiration of amended soil to a two-pool model. After 2, 7 and 30 days of incubation, the control and amended soils were sampled and analyzed for their content of extractable organic C, N, NO3- and NH4+ and microbial biomass C and N. Results showed that SCG by-products presented a great variability in their properties. SCG and hydrochars presented higher contents in volatile matter and water soluble C and N, and low content of fixed C, while biochars showed an opposite behaviour. SEM images confirmed the different characteristics of the SCG by-products: the biochar presented a porous structure, honeycomb-like form, due to the loss of the more soluble compounds, while the SCG and hydrochars' pores were filled with amorphous carbonaceous materials. Consequently, soil addition of SCG by-products showed a distinct impact on C and N cycle and microbial biomass content. Addition of SCG and hydrochars generated the highest cumulative CO2-C emissions (2103-2300 µg g-1), the lower amount of CR (86.8-88.6%), increased the soil extractable organic C and microbial biomass C and N and caused N immobilization. On the other hand, the addition of biochars generated lower CO2-C emissions (542-1060 µg g-1), higher amounts of CR (96. 3-99.9%) and lower amounts of extractable compounds and microbial biomass C and N, generating also N immobilization, but to a lesser extent. The addition of vermicompost generated 723 µg g-1 of CO2-C and 98% of CR remaining. However, this by-product did not generate N immobilization being able to act as N fertilizer. None of the residues generated N2O emissions. The different properties of the SCG by-products and their impact on C and N cycle indicated that they can be effectively applied to soil to exert different agronomical and environmental functions.

Post-digestate composting benefits and the role of enzyme activity to predict trace element immobilization and compost maturity.

The current study evaluated the quality of agricultural waste digestate by composting or co-composting with biogas feedstock (maize silage, food processing waste, or poultry litter). Temperature, phytotoxicity, C/N ratio, water extractable trace elements, and 14 enzyme activities were monitored. Temperature dropped earlier in digestate and maize silage co-composting pile, reducing time to maturity by 20 days. Composting and co-composting reduced phytotoxicity and C/N ratio, but increased immobilization of Al, Ba, Fe, Zn, and Mn at least by 40% in all piles. All the enzyme activities, except arylsulfatase and α-glucosidase, increased at the maturity phase and negatively correlated with organic matter content and most of trace elements. Post-digestate composting or co-composting with biogas feedstock is a promising strategy to improve digestate quality for fertilizer use, and selected enzyme activities can be indicators of compost maturity and immobilization of trace elements.

Bioaugmented Phytoremediation of Metal-Contaminated Soils and Sediments by Hemp and Giant Reed.

We assessed the effects of EDTA and selected plant growth-promoting rhizobacteria (PGPR) on the phytoremediation of soils and sediments historically contaminated by Cr, Ni, and Cu. A total of 42 bacterial strains resistant to these heavy metals (HMs) were isolated and screened for PGP traits and metal bioaccumulation, and two Enterobacter spp. strains were finally selected. Phytoremediation pot experiments of 2 months duration were carried out with hemp (Cannabis sativa L.) and giant reed (Arundo donax L.) grown on soils and sediments respectively, comparing in both cases the effects of bioaugmentation with a single PGPR and EDTA addition on plant and root growth, plant HM uptake, HM leaching, as well as the changes that occurred in soil microbial communities (structure, biomass, and activity). Good removal percentages on a dry mass basis of Cr (0.4%), Ni (0.6%), and Cu (0.9%) were observed in giant reed while negligible values (<100‰) in hemp. In giant reed, HMs accumulated differentially in plant (rhizomes > > roots > leaves > stems) with largest quantities in rhizomes (Cr 0.6, Ni 3.7, and Cu 2.2 g plant-1). EDTA increased Ni and Cu translocation to aerial parts in both crops, despite that in sediments high HM concentrations in leachates were measured. PGPR did not impact fine root diameter distribution of both crops compared with control while EDTA negatively affected root diameter class length (DCL) distribution. Under HM contamination, giant reed roots become shorter (from 5.2 to 2.3 mm cm-3) while hemp roots become shorter and thickened from 0.13 to 0.26 mm. A consistent indirect effect of HM levels on the soil microbiome (diversity and activity) mediated by plant response (root DCL distribution) was observed. Multivariate analysis of bacterial diversity and activity revealed not only significant effects of plant and soil type (rhizosphere vs. bulk) but also a clear and similar differentiation of communities between control, EDTA, and PGPR treatments. We propose root DCL distribution as a key plant trait to understand detrimental effect of HMs on microbial communities. Positive evidence of the soil-microbe-plant interactions occurring when bioaugmentation with PGPR is associated with deep-rooting perennial crops makes this combination preferable over the one with chelating agents. Such knowledge might help to yield better bioaugmented bioremediation results in contaminated sites.

With or Without You: Altered Plant Response to Boron-Deficiency in Hydroponically Grown Grapevines Infected by Grapevine Pinot Gris Virus Suggests a Relation Between Grapevine Leaf Mottling and Deformation Symptom Occurrence and Boron Plant Availability.

Despite the increasing spread of Grapevine Leaf Mottling and Deformation (GLMD) worldwide, little is known about its etiology. After identification of grapevine Pinot gris virus (GPGV) as the presumptive causal agent of the disease in 2015, various publications have evaluated GPGV involvement in GLMD. Nevertheless, there are only partial clues to explain the presence of GPGV in both symptomatic and asymptomatic grapevines and the mechanisms that trigger symptom development, and so a consideration of new factors is required. Given the similarities between GLMD and boron (B)-deficiency symptoms in grapevine plants, we posited that GPGV interferes in B homeostasis. By using a hydroponic system to control B availability, we investigated the effects of different B supplies on grapevine phenotype and those of GPGV infection on B acquisition and translocation machinery, by means of microscopy, ionomic and gene expression analyses in both roots and leaves. The transcription of the genes regulating B homeostasis was unaffected by the presence of GPGV alone, but was severely altered in plants exposed to both GPGV infection and B-deficiency, allowing us to speculate that the capricious and patchy occurrence of GLMD symptoms in the field may not be related solely to GPGV, but to GPGV interference in plant responses to different B availabilities. This hypothesis found preliminary positive confirmations in analyses on field-grown plants.

Effects of Seed-Applied Biofertilizers on Rhizosphere Biodiversity and Growth of Common Wheat (Triticum aestivum L.) in the Field.

In order to reduce chemical fertilization and improve the sustainability of common wheat (Triticum aestivum L.) cultivation, maintaining at the same time high production and quality standards, this study investigated the effects of three commercial biofertilizers on rhizosphere bacterial biomass, biodiversity and enzymatic activity, and on plant growth and grain yield in a field trial. The wheat seeds were inoculated with the following aiding microrganisms: (i) a bacterial consortium (Azospirillum spp. + Azoarcus spp. + Azorhizobium spp.); and two mycorrhizal fungal-bacterial consortia, viz. (ii) Rhizophagus irregularis + Azotobacter vinelandii, and (iii) R. irregularis + Bacillus megaterium + Frateuria aurantia, and comparisons were made with noninoculated controls. We demonstrate that all the biofertilizers significantly enhanced plant growth and nitrogen accumulation during stem elongation and heading, but this was translated into only small grain yield gains (+1%-4% vs controls). The total gluten content of the flour was not affected, but in general biofertilization significantly upregulated two high-quality protein subunits, i.e., the 81 kDa high-molecular-weight glutenin subunit and the 43.6 kDa low-molecular-weight glutenin subunit. These effects were associated with increases in the rhizosphere microbial biomass and the activity of enzymes such as ß-glucosidase, α-mannosidase, ß-mannosidase, and xylosidase, which are involved in organic matter decomposition, particularly when Rhizophagus irregularis was included as inoculant. No changes in microbial biodiversity were observed. Our results suggest that seed-applied biofertilizers may be effectively exploited in sustainable wheat cultivation without altering the biodiversity of the resident microbiome, but attention should be paid to the composition of the microbial consortia in order to maximize their benefits in crop cultivation.

Spectrophotometric methods for the measurement of soil pH: A reappraisal.

The pH of aqueous soil extracts is generally measured potentiometrically by glass electrode (GE). Spectrophotometric methods have also been used till around the '80s, and then they have apparently been abandoned. However, the use of microplates and spectrophotometers able to read absorbance values in them can significantly increase the speed of the data collection (saving analysis time), thus justifying their reappraisal. Three spectrophotometric methods are proposed in this work: a one-indicator (OISM), a separated three-indicator (STISM) and a mixed three-indicator (MTISM) spectrophotometric method. They are based on the addition of one or three colorimetric indicators (methyl red, bromocresol violet, and bromothymol blue) to the aqueous extract. The pH is measured through its absorption properties in the visible region. The analysis of 60 soil samples showed that STISM and MTISM results correlate well with the ones obtained by GE. The STISM method, being more simple and general than the other two methods, is proposed for quick routine analyses. The repeatability, reproducibility and accuracy of STISM (and of GE, for comparison) were evaluated by measuring several times the pH of a series of soil samples and of a certified reference soil. The standard deviations of STISM results were slightly worse than those of GE, whereas the accuracy was slightly better, indicating that STISM and GE have overall similar performances. STISM method is much faster than GE one: the analysis time saving is around 2 min per sample, and it becomes very high when hundreds of samples have to be analysed. It follows that STISM can represent an advantageous alternative to GE for rapid and accurate soil pH measurements.

Microbial Decomposer Dynamics: Diversity and Functionality Investigated through a Transplantation Experiment in Boreal Forests.

Litter decomposition is the main source of mineral nitrogen (N) in terrestrial ecosystem and a key step in carbon (C) cycle. Microbial community is the main decomposer, and its specialization on specific litter is considered at the basis of higher decomposition rate in its natural environment than in other forests. However, there are contrasting evidences on how the microbial community responds to a new litter input and if the mass loss is higher in natural environment. We selected leaf litter from three different plant species across three sites of different altitudinal ranges: oak (Quercus petraea (Matt.) Liebl., 530 m a.s.l), beech (Fagus sylvatica L., 1000 m a.s.l.), rhododendron (Rhododendron ferrugineum L., 1530 m a.s.l.). A complete transplantation experiment was set up within the native site and the other two altitudinal sites. Microbial community structure was analyzed via amplified ribosomal intergenic spacer analysis (ARISA) fingerprinting. Functionality was investigated by potential enzyme activities. Chemical composition of litter was recorded. Mass loss showed no faster decomposition rate on native site. Similarly, no influence of site was found on microbial structure, while there was a strong temporal variation. Potential enzymatic activities were not affected by the same temporal pattern with a general increase of activities during autumn. Our results suggested that no specialization in microbial community is present due to the lack of influence of the site in structure and in the mass loss dynamics. Finally, different temporal patterns in microbial community and potential enzymatic activities suggest the presence of functional redundancy within decomposers.

Biochar lowers ammonia emission and improves nitrogen retention in poultry litter composting.

The poultry industry produces abundant quantities of nutrient-rich litter, much of which is composted before use as a soil amendment. However, a large proportion of nitrogen (N) in poultry litter is lost via volatilisation during composting, with negative environmental and economic consequences. This study examined the effect of incorporating biochar during composting of poultry litter on ammonia (NH3) volatilisation and N retention. Biochars produced at 550°C from greenwaste (GWB) and poultry litter (PLB) feedstocks were co-composted with a mixture of raw poultry litter and sugarcane straw [carbon (C):N ratio 10:1] in compost bins. Ammonia emissions accounted for 17% of the total N (TN) lost from the control and 12-14% from the biochar-amended compost. The TN emitted as NH3, as a percentage of initial TN, was significantly lower (P<0.05) i.e. by 60% and 55% in the compost amended with GWB and PLB, respectively, relative to the control. The proportion of N retained in the finished compost, as a percentage of initial TN, was 84%, 78% and 67% for the GWB, PLB and nil biochar control, respectively. Lower concentration of dissolved organic C (DOC) together with higher activity of beta-glucosidase and leucine-aminopeptidase were found in the GWB-amended compost (cf. control). It is hypothesized that lower NH3 emission in the GWB-amended compost was caused not just by the higher surface area of this biochar but could also be related to greater incorporation of ammonium (NH4+) in organic compounds during microbial utilisation of DOC. Furthermore, the GWB-amended compost retained more NH4+ at the end of composting than the PLB-amended compost. Results showed that addition of biochar, especially GWB, generated multiple benefits in composting of poultry litter: decrease of NH3 volatilisation, decrease in NH3 toxicity towards microorganisms, and improved N retention, thus enhancing the fertiliser value of the composted litter. It is suggested that the latter benefit is linked to a beneficial modification of the microbial environment.

Effects of slope exposure on soil physico-chemical and microbiological properties along an altitudinal climosequence in the Italian Alps.

Due to their sensitivity to changing environmental conditions sub- and alpine soils are often monitored in the context of climate change, usually, however, neglecting slope exposure. Therefore, we set up a climosequence-approach to study the effect of exposure and, in general, climate, on the microbial biomass and microbial diversity and activity, comprising five pairs of north (N)- and south (S)-facing sites along an altitudinal gradient ranging from 1200 to 2400m a.s.l. in the Italian Alps (Trentino Alto Adige, Italy). Soil physico-chemical properties were related to microbiological properties (microbial biomass: double strand DNA yield vs. substrate-induced respiration; diversity of bacterial, fungal and archaeal communities: genetic fingerprinting DGGE vs. real-time PCR; microbial activity: basal respiration vs. multiple hydrolytic enzyme assays) to monitor shifts in the diversity and activity of microbial communities as a function of slope exposure and to evaluate the most determinant chemical parameters shaping the soil microbiota. The exposure-effect on several hydrolytic key-enzymes was enzyme-specific: e.g. acid phosphomonoesterase potential activity was more pronounced at the N-facing slope while the activities of alkaline phosphomonoesterase, pyrophosphate-phosphodiesterase and arylsulfatase were higher at the S-facing slope. Furthermore, this exposure-effect was domain-specific: bacteria (S>N, altitude-independent); fungi (N~S); and archaea (N>S; altitude-dependent). Additionally, the abiotic parameters shaping the community composition were in general depending on soil depth. Our multidisciplinary approach allowed us to survey the exposure and altitudinal effects on soil physico-chemical and microbiological properties and thus unravel the complex multiple edaphic factor-effects on soil microbiota in mountain ecosystems.

End-product inhibition and acidification limit biowaste fermentation efficiency.

Converting waste to resource may mitigate environmental pollution and global resource limitation. The platform chemical lactic acid can be produced from biowaste and its liquid fraction after solid-liquid separation. A fermentation step for lactic acid production prior to the conversion of biowaste to methane and organic fertilizer would increase the biowaste's value. Despite the huge potential and promising results of the treatment procedure, the reasons for efficiency loss observed previously need to be addressed in order to pave the way for an up-scaling of the fermentation process. Therefore, biowaste was fermented applying pH control, acid extraction and glucose addition in order to counteract reasons such as acidification, end-product inhibition and carbon limitation, respectively. The fermentation was competitive compared to other renewable lactic acid production substrates and reached a maximum productivity of >5 g Clactic acidg(-1)Ch(-1) and a concentration exceeding 30 g L(-1). A combination of acidification and end-product inhibition was identified as major obstacle. Lactobacillus crispatus and its closest relatives were identified as key lactic acid producers within the process using Miseq Illumina sequencing.

Microbiological features and bioactivity of a fermented manure product (preparation 500) used in biodynamic agriculture.

The fermented manure derivative known as Preparation 500 is traditionally used as a field spray in biodynamic agriculture for maintaining and increasing soil fertility. This work aimed at characterizing the product from a microbiological standpoint and at assaying its bioactive properties. The approach involved molecular taxonomical characterization of the culturable microbial community; ARISA fingerprints of the total bacteria and fungal communities; chemical elemental macronutrient analysis via a combustion analyzer; activity assays for six key enzymes; bioassays for bacterial quorum sensing and chitolipooligosaccharide production; and plant hormonelike activity. The material was found to harbor a bacterial community of 2.38 × 10(8) CFU/g dw dominated by Grampositives with minor instances of Actinobacteria and Gammaproteobacteria. ARISA showed a coherence of bacterial assemblages in different preparation lots of the same year in spite of geographic origin. Enzymatic activities showed elevated values of beta-glucosidase, alkaline phosphatase, chitinase, and esterase. The preparation had no quorum sensing-detectable signal, and no rhizobial nod gene-inducing properties, but displayed a strong auxin-like effect on plants. Enzymatic analyses indicated a bioactive potential in the fertility and nutrient cycling contexts. The IAA activity and microbial degradation products qualify for a possible activity as soil biostimulants. Quantitative details and possible modes of action are discussed.