The community of root fungi is associated with the growth rate of Norway spruce (Picea abies).

Our study delved into the relationship between root-associated fungi, gene expression and plant morphology in Norway spruce cuttings derived from both slow-and fast-growing trees. We found no clear link between the gene expression patterns of adventitious roots and the growth phenotype, suggesting no fundamental differences in the receptiveness to fungal symbionts between the phenotypes. Interestingly, saplings from slow-growing parental trees exhibited a higher richness of ectomycorrhizal species and larger roots. Some ectomycorrhizal species, typically found on mature spruces, were more prevalent on saplings from slow-growing spruces. The ericoid mycorrhizal fungus, Hyaloscypha hepaticola, showed a stronger association with saplings from fast-growing spruces. Moreover, saplings from slow-growing spruces had a greater number of Ascomycete taxa and free-living saprotrophic fungi. Aboveground sapling stems displayed some phenotypic variation; saplings from fast-growing phenotypes had longer branches but fewer whorls in their stems compared to those from the slow-growing group. In conclusion, the observed root-associated fungi and phenotypic characteristics in young Norway spruces may play a role in their long-term growth rate. This suggests that the early interactions between spruces and fungi could potentially influence their growth trajectory.

Impact of Heterobasidion root-rot on fine root morphology and associated fungi in Picea abies stands on peat soils.

We examined differences in fine root morphology, mycorrhizal colonisation and root-inhabiting fungal communities between Picea abies individuals infected by Heterobasidion root-rot compared with healthy individuals in four stands on peat soils in Latvia. We hypothesised that decreased tree vitality and alteration in supply of photosynthates belowground due to root-rot infection might lead to changes in fungal communities of tree roots. Plots were established in places where trees were infected and in places where they were healthy. Within each stand, five replicate soil cores with roots were taken to 20 cm depth in each root-rot infected and uninfected plot. Root morphological parameters, mycorrhizal colonisation and associated fungal communities, and soil chemical properties were analysed. In three stands root morphological parameters and in all stands root mycorrhizal colonisation were similar between root-rot infected and uninfected plots. In one stand, there were significant differences in root morphological parameters between root-rot infected versus uninfected plots, but these were likely due to significant differences in soil chemical properties between the plots. Sequencing of the internal transcribed spacer of fungal nuclear rDNA from ectomycorrhizal (ECM) root morphotypes of P. abies revealed the presence of 42 fungal species, among which ECM basidiomycetes Tylospora asterophora (24.6 % of fine roots examined), Amphinema byssoides (14.5 %) and Russula sapinea (9.7 %) were most common. Within each stand, the richness of fungal species and the composition of fungal communities in root-rot infected versus uninfected plots were similar. In conclusion, Heterobasidion root-rot had little or no effect on fine root morphology, mycorrhizal colonisation and composition of fungal communities in fine roots of P. abies growing on peat soils.

The ectomycorrhizal community of conifer stands on peat soils 12 years after fertilization with wood ash.

We studied long-term effects of fertilization with wood ash on biomass, vitality and mycorrhizal colonization of fine roots in three conifer forest stands growing in Vacciniosa turf. mel. (V), Myrtillosa turf. mel. (M) and Myrtillosa turf. mel./Caricoso-phragmitosa (MC) forest types on peat soils. Fertilization trials amounting 5 kg/m(2) of wood ash were established 12 years prior to this study. A total of 63 soil samples with roots were collected and analysed. Ectomycorrhizal (ECM) fungi in roots were identified by morphotyping and sequencing of the fungal internal transcribed spacer (ITS) region. In all forest types, fine root biomass was higher in fertilized plots than in control plots. In M forest type, proportion of living fine roots was greater in fertilized plots than in control plots, while in V and MC, the result was opposite. Fifty ECM species were identified, of which eight were common to both fertilized and control plots. Species richness and Shannon diversity index were generally higher in fertilized plots than in control plots. The most common species in fertilized plots were Amphinema byssoides (17.8%) and Tuber cf. anniae (12.2%), while in control plots, it was Tylospora asterophora (18.5%) and Lactarius tabidus (20.3%). Our results showed that forest fertilization with wood ash has long-lasting effect on diversity and composition of ECM fungal communities.

Profiling functions of ectomycorrhizal diversity and root structuring in seedlings of Norway spruce (Picea abies) with fast- and slow-growing phenotypes.

We studied the role of taxonomical and functional ectomycorrhizal (ECM) fungal diversity in root formation and nutrient uptake by Norway spruce (Picea abies) seedlings with fast- and slow-growing phenotypes. Seedlings were grown with an increasing ECM fungal diversity gradient from one to four species and sampled before aboveground growth differences between the two phenotypes were apparent. ECM fungal colonization patterns were determined and functional diversity was assayed via measurements of potential enzyme activities of eight exoenzymes probably involved in nutrient mobilization. Phenotypes did not vary in their receptiveness to different ECM fungal species. However, seedlings of slow-growing phenotypes had higher fine-root density and thus more condensed root systems than fast-growing seedlings, but the potential enzyme activities of ectomycorrhizas did not differ qualitatively or quantitatively. ECM species richness increased host nutrient acquisition potential by diversifying the exoenzyme palette. Needle nitrogen content correlated positively with high chitinase activity of ectomycorrhizas. Rather than fast- and slow-growing phenotypes exhibiting differing receptiveness to ECM fungi, our results suggest that distinctions in fine-root structuring and in the belowground growth strategy already apparent at early stages of seedling development may explain later growth differences between fast- and slow-growing families.

Comments to "Degli-Innocenti, F. The pathology of hype, hyperbole and publication bias is creating an unwarranted concern towards biodegradable mulch films" [J. Hazard. Mater. 463 (2024) 132923].

Some narratives present biodegradable plastic use for soil mulching practices in agriculture as "environmentally friendly" and "sustainable" alternatives to conventional plastics. To verify these narratives, environmental research recently started focusing on their potential impact on soil health, highlighting some concerns. The paper by Degli-Innocenti criticizes this unfolding knowledge arguing that it is affected by communication hypes, alarmistic writing and a focus on exposure scenarios purposedly crafted to yield negative effects. The quest of scientists for increased impact - the paper concludes - is the driver of such behavior. As scholars devoted to the safeguarding of scientific integrity, we set to verify whether this serious claim is grounded in evidence. Through a bibliometric analysis (using number of paper reads, citations and mentions on social media to measure the impact of publications) we found that: i) the papers pointed out by Degli-Innocenti as examples of biased works do not score higher than the median of similar publications; ii) the methodology used to support the conclusion is non-scientific; and iii) the paper does not fulfil the requirements concerning disclosure of conflicts of interests. We conclude that this paper represents a non-scientific opinion, potentially biased by a conflict of interest. We ask the paper to be clearly tagged as such, after the necessary corrections on the ethic section have been made. That being said, the paper does offer some useful insights for the definition of exposure scenarios in risk assessment. We comment and elaborate on these proposed models, hoping that this can help to advance the field.

Comparing the impact of microplastics derived from a biodegradable and a conventional plastic mulch on plant performance.

Agricultural lands have been identified as plastic sinks. One source is plastic mulches, which are a source of micro- and nano-sized plastics in agricultural soils. Because of their persistence, there is now a push towards developing biodegradable plastics, which are designed to undergo (partial) breakdown after entering the environment. Yet, limited research has investigated the impacts of both conventional and biodegradable plastics on distinct plants. Moreover, comparisons among studies are difficult due to differences in experimental design. This study directly compares the effects of artificially weathered conventional polyethylene (PE) and starch-based biodegradable polybutylene adipate terephthalate (PBAT) on four food crops, including two monocots (barley, Hordeum vulgare, and wheat, Triticum aestivum L.) and two dicots (carrot, Daucus carota, and lettuce, Lactuca sativa L.). We investigated the effects of environmentally relevant low, medium, and high (0.01 %, 0.1 %, 1 % w/w) concentrations of PE and starch-PBAT blend on seed germination (acute toxicity), and subsequently on plant growth and chlorophyll through a pot-plant experiment (chronic toxicity). Germination of all species was not affected by both plastics. However, root length was reduced for lettuce and wheat seedlings. No other effects were recorded on monocots. We observed a reduction in shoot length and bud wet weight of carrot seedlings for the highest concentration of PE and starch-PBAT blend. Chronic exposure resulted in a significant decrease in shoot biomass of barley and lettuce. Additionally, a positive increase in the number of leaves of lettuce was observed for both plastics. Chlorophyll content was increased in lettuce when exposed to PE and starch-PBAT blend. Overall, adverse effects in dicots were more abundant than in monocots. Importantly, we found that the biodegradable plastic caused more commonly adverse effects on plants compared to conventional plastic, which was confirmed by a mini-review of studies directly comparing the impact of conventional and biodegradable microplastics.

Impacts of coniferous bark-derived organic soil amendments on microbial communities in arable soil - a microcosm study.

A decline in the carbon content of agricultural soils has been reported globally. Amendments of forest industry side-streams might counteract this. We tested the effects of industrial conifer bark and its cascade process materials on the soil microbiome under barley (Hordeum vulgare L.) in clay and silt soil microcosms for 10 months, simulating the seasonal temperature changes of the boreal region. Microbial gene copy numbers were higher in clay soils than in silt. All amendments except unextracted bark increased bacterial gene copies in both soils. In turn, all other amendments, but not unextracted bark from an anaerobic digestion process, increased fungal gene copy numbers in silt soil. In clay soil, fungal increase occurred only with unextracted bark and hot water extracted bark. Soil, amendment type and simulated season affected both the bacterial and fungal community composition. Amendments increased bacteria originating from the anaerobic digestion process, as well as dinitrogen fixers and decomposers of plant cells. In turn, unextracted and hot water extracted bark determined the fungal community composition in silt. As fungal abundance increase and community diversification are related to soil carbon acquisition, bark-based amendments to soils can thus contribute to sustainable agriculture.

Fast and reliable method to estimate global DNA methylation in plants and fungi with high-pressure liquid chromatography (HPLC)-ultraviolet detection and even more sensitive one with HPLC-mass spectrometry.

DNA (Deoxyribonucleic acid) methylation is one of the epigenetic modifications of DNA, acting as a bridge between genotype and phenotype. Thus, disruption of DNA methylation pattern has tremendous consequences for organism development. Current methods to determine DNA methylation suffer from methodological drawbacks like high requirement of DNA and poor reproducibility of chromatograms. Here we provide a fast and reliable method using high-pressure liquid chromatography (HPLC)-ultraviolet (UV) detector and even more sensitive one with HPLC- mass spectrometry (MS) and we test this method with various plant and fungal DNA isolates. We optimized the preparation of the DNA degradation step to decrease background noise, we improved separation conditions to provide reliable and reproducible chromatograms and conditions to measure nucleotides in HPLC-MS. We showed that global DNA methylation level can be accurately and reproducibly measured with as little as 0.2 µM for HPLC-UV and 0.02 µM for HPLC-MS of methylated cytosine.

Species-dependent responses of crop plants to polystyrene microplastics.

Only recently there has been a strong focus on the impacts of microplastics on terrestrial crop plants. This study aims to examine and compare the effects of microplastics on two monocotyledonous (barley, Hordeum vulgare and wheat, Triticum aestivum), and two dicotyledonous (carrot, Daucus carota and lettuce, Lactuca sativa) plant species through two complimentary experiments. First, we investigated the effects of low, medium, and high (103, 105, 107 particles per mL) concentrations of 500 nm polystyrene microplastics (PS-MPs) on seed germination and early development. We found species-dependent effects on the early development, with microplastics only significantly affecting lettuce and carrot. When acutely exposed during germination, PS-MPs significantly delayed the germination of lettuce by 24%, as well as promoted the shoot growth of carrot by 71% and decreased its biomass by 26%. No effect was recorded on monocot species. Secondly, we performed a chronic (21 d) hydroponic experiment on lettuce and wheat. We observed that PS-MPs significantly reduced the shoot growth of lettuce by up to 35% and increased its biomass by up to 64%, while no record was reported on wheat. In addition, stress level indicators and defence mechanisms were significantly up-regulated in both lettuce and wheat seedlings. Overall, this study shows that PS-MPs affect plant development: impacts were recorded on both germination and growth for dicots, and responses identified by biochemical markers of stress were increased in both lettuce and wheat. This highlights species-dependent effects as the four crops were grown under identical conditions to allow direct comparison. For future research, our study emphasizes the need to focus on crop specific effects, while also working towards knowledge of plastic-induced impacts at environmentally relevant conditions.

Polystyrene nanoparticles induce concerted response of plant defense mechanisms in plant cells.

Recent advances in knowledge suggest that micro- and nanoplastics pose a threat to plant health, however, the responses of plants to this stressor are not well-known. Here we examined the response of plant cell defence mechanisms to nanoparticles of commonly used plastic, polystyrene. We used plant cell cultures of widely cultivated plants, the monocots wheat and barley (Triticum aestivum L., Hordeum vulgare L.) and the dicots carrot and tomato (Daucus carota L., Solanum lycopersicum L.). We measured the activities of enzymes involved in the scavenging of reactive oxygen species and nonenzymatic antioxidants and we estimated potential damages in plant cell structures and functioning via lipid peroxidation and DNA methylation levels. Our results demonstrate that the mode of action of polystyrene nanoparticles on plant cells involves oxidative stress. However, the changes in plant defence mechanisms are dependent on plant species, exposure time and nanoplastic concentrations. In general, both monocots showed similar responses to nanoplastics, but the carrot followed more the response of monocots than a second dicot, a tomato. Higher H2O2, lipid peroxidation and lower enzyme activities scavenging H2O2 suggest that tomato cells may be more susceptible to polystyrene-induced stress. In conclusion, polystyrene nanoplastics induce oxidative stress and the response of the plant defense mechanisms involving several chain reactions leading to oxidoreductive homeostasis.

Pulp and paper mill sludges decrease soil erodibility.

Declining carbon (C) content in agricultural soils threatens soil fertility and makes soil prone to erosion, which could be rectified with organic soil amendments. In a 4-yr field trial, we made a single application of three different organic sludges from the pulp and paper industry and studied their effects on cereal yield, soil C content, and fungal and bacterial composition. In laboratory rainfall simulations, we also studied the effects of the soil amendments on susceptibility to erosion and nutrient mobilization of a clay-textured soil by measuring the quality of percolation water passing through 40-cm intact soil monoliths during 2-d rainfall simulations over four consecutive years after application. A nutrient-poor fiber sludge reduced wheat yield in the first growing season, but there were no other significant effects on cereal yield or grain quality. An input of ∼8 Mg ha-1 C with the soil amendments had only minor effects on soil C content after 4 yr, likely because of fast microbe-mediated turnover. The amendments clearly changed the fungal and bacterial community composition. All amendments significantly reduced suspended solids (SS) and total phosphorus (TP) concentrations in percolation water. The effect declined with time, but the reduction in SS and TP was still >25% 4 yr after application. We attributed the lower tendency for particle detachment in rain simulations to direct interactions of soil minerals with the added particulate organic matter and microbe-derived compounds that stabilize soil aggregates. In soils with low organic matter content, pulp and paper industry by-products can be a viable measure for erosion mitigation.

Are there environmental or agricultural benefits in using forest residue biochar in boreal agricultural clay soil?

Short-term agronomic and environmental benefits are fundamental factors in encouraging farmers to use biochar on a broad scale. The short-term impacts of forest residue biochar (BC) on the productivity and carbon (C) storage of arable boreal clay soil were studied in a field experiment. In addition, rain simulations and aggregate stability tests were carried out to investigate the potential of BC to reduce nutrient export to surface waters. A BC addition of 30 t ha-1 increased soil test phosphorus and decreased bulk density in the surface soil but did not significantly change pH or water retention properties, and most importantly, did not increase the yield. There were no changes in the bacterial or fungal communities, or biomasses. Soil basal respiration was higher in BC-amended plots in the spring, but no differences in respiration rates were detected in the fall two years after the application. Rain simulation experiments did not support the use of BC in reducing erosion or the export of nutrients from the field. Of the C added, on average 80% was discovered in the 0-45 cm soil layer one year after the application. Amendment of boreal clay soil with a high rate of BC characterized by a moderately alkaline pH, low surface functionalities, and a recalcitrant nature, did not induce such positive impacts that would unambiguously motivate farmers to invest in BC. BC use seems unviable from the farmer's perspective but could play a role in climate change mitigation, as it will likely serve as long-term C storage.

Early root growth and architecture of fast- and slow-growing Norway spruce (Picea abies) families differ-potential for functional adaptation.

The relationship between the growth rate of aboveground parts of trees and fine root development is largely unknown. We investigated the early root development of fast- and slow-growing Norway spruce (Picea abies (L.) H. Karst.) families at a developmental stage when the difference in size is not yet observed. Seedling root architecture data, describing root branching, were collected with the WinRHIZO™ image analysis system, and mixed models were used to determine possible differences between the two growth phenotypes. A new approach was used to investigate the spatial extent of root properties along the whole sample root from the base of 1-year-old seedlings to the most distal part of a root. The root architecture of seedlings representing fast-growing phenotypes showed ~30% higher numbers of root branches and tips, which resulted in larger root extensions and potentially a better ability to acquire nutrients. Seedlings of fast-growing phenotypes oriented and allocated root tips and biomass further away from the base of the seedling than those growing slowly, a possible advantage in nutrient-limited and heterogeneous boreal forest soils. We conclude that a higher long-term growth rate of the aboveground parts in Norway spruce may relate to greater allocation of resources to explorative roots that confers a competitive edge during early growth phases in forest ecosystems.

The community of needle endophytes reflects the current physiological state of Norway spruce.

This study investigated fungal endophytes in the needles of Norway spruce (Picea abies) cuttings in relation to host tree growth. We also determined the prevalence of endophytes in needles incubated for six months. The cuttings originated from clonal origins showing slow- and fast-growth in long-term field trials but the heritable differences in growth rate were not yet detected among the studied cutting. Endophytes were isolated from surface-sterilized needles with culture-free DNA techniques. No significant differences were observed between endophyte communities of slow- and fast-growing clonal origins. However, the endophyte community correlated with the current growth rate of cuttings suggesting that endophytes reflect short- rather than long-term performance of a host. The concentration of condensed tannins was similar in slow- and fast-growing clonal origins but it showed a negative relationship with endophyte species richness, implying that these secondary compounds may play an important role in spruce tolerance against fungal infections. More than a third of endophyte species were detected in both fresh and decomposing needles, indicating that many needle endophytes are facultative saprotrophs. Several potentially pathogenic fungal species were also found within the community of saprotrophic endophytes.

Endophyte communities vary in the needles of Norway spruce clones.

Endophytic fungi show no symptoms of their presence but can influence the performance and vitality of host trees. The potential use of endophytes to indicate vitality has been previously realized, but a standard protocol has yet to be developed due to an incomplete understanding of the factors that regulate endophyte communities. Using a culture-free molecular approach, we examined the extent to which host genotype influences the abundance, species richness, and community composition of endophytic fungi in Norway spruce needles. Briefly, total DNA was extracted from the surface-sterilized needles of 30 clones grown in a nursery field and the copy number of the fungal internal transcribed spacer (ITS) region of ribosomal DNA was estimated by quantitative PCR. Fungal species richness and community composition were determined by denaturing gradient gel electrophoresis and DNA sequencing. We found that community structure and ITS copy number varied among spruce clones, whereas species richness did not. Host traits interacting with endophyte communities included needle surface area and the location of cuttings in the experimental area. Although Lophodermium piceae is considered the dominant needle endophyte of Norway spruce, we detected this species in only 33% of samples. The most frequently observed fungus (66%) was the potentially pathogenic Phoma herbarum. Interestingly, ITS copy number of endophytic fungi correlated negatively with the richness of ectomycorrhizal fungi and thus potential interactions between fungal communities and their influence on the host tree are discussed. Our results suggest that in addition to environmental factors, endophyte communities of spruce needles are determined by host tree identity and needle surface area.