Root uptake of cereal benzoxazinoids grants resistance to root-knot nematode invasion in white clover.

Plants synthesize a plethora of chemical defence compounds, which vary between evolutionary lineages. We hypothesize that plants evolved the ability to utilize defence compounds synthesized and released by neighbouring heterospecific plants. In two experiments, we incubated clover (Trifolium repens L.) seedlings with individual benzoxazinoid (BX) compounds (2,4-dihydroxy-1,4-benzoxazin-3-one, 2-hydroxy-1,4-benzoxazin-3-one, benzoxazolinone, and 6-methoxy- benzoxazolin-2-one), a group of bioactive compounds produced by cereals, to allow clover BX uptake. Subsequently, we transplanted the seedlings into soil and quantified BX root and shoot content and invasion of root-knot nematodes in clover roots up to 8 weeks after transplantation. We show that clover root uptake of BXs substantially enhanced clover's resistance against the root-knot nematode Meloidogyne incognita. This effect lasted up to 6 weeks after the clover roots were exposed to the BXs. BXs were absorbed by clover roots, and then translocated to the shoots. As a result of clover metabolization, we detected the parent BXs and a range of their transformation products in the roots and shoots. Based on these novel findings, we envisage that co-cultivation of crop species with complementary and transferable chemical defence systems can add to plant protection.

Early assessment of fungal and oomycete pathogens in greenhouse irrigation water using Oxford nanopore amplicon sequencing.

Water-borne plant pathogenic fungi and oomycetes are a major threat in greenhouse production systems. Early detection and quantification of these pathogens would enable us to ascertain both economic and biological thresholds required for a timely treatment, thus improving effective disease management. Here, we used Oxford nanopore MinION amplicon sequencing to analyze microbial communities in irrigation water collected from greenhouses used for growing tomato, cucumber and Aeschynanthus sp. Fungal and oomycete communities were characterized using primers that amplify the full internal transcribed spacer (ITS) region. To assess the sensitivity of the MinION sequencing, we spiked serially diluted mock DNA into the DNA isolated from greenhouse water samples prior to library preparation. Relative abundances of fungal and oomycete reads were distinct in the greenhouse irrigation water samples and in water samples from setups with tomato that was inoculated with Fusarium oxysporum. Sequence reads derived from fungal and oomycete mock communities were proportionate in the respective serial dilution samples, thus confirming the suitability of MinION amplicon sequencing for environmental monitoring. By using spike-ins as standards to test the reliability of quantification using the MinION, we found that the detection of spike-ins was highly affected by the background quantities of fungal or oomycete DNA in the sample. We observed that spike-ins having shorter length (538bp) produced reads across most of our dilutions compared to the longer spikes (>790bp). Moreover, the sequence reads were uneven with respect to dilution series and were least retrievable in the background samples having the highest DNA concentration, suggesting a narrow dynamic range of performance. We suggest continuous benchmarking of the MinION sequencing to improve quantitative metabarcoding efforts for rapid plant disease diagnostic and monitoring in the future.

Harnessing root-soil-microbiota interactions for drought-resilient cereals.

Cereal plants form complex networks with their associated microbiome in the soil environment. A complex system including variations of numerous parameters of soil properties and host traits shapes the dynamics of cereal microbiota under drought. These multifaceted interactions can greatly affect carbon and nutrient cycling in soil and offer the potential to increase plant growth and fitness under drought conditions. Despite growing recognition of the importance of plant microbiota to agroecosystem functioning, harnessing the cereal root microbiota remains a significant challenge due to interacting and synergistic effects between root traits, soil properties, agricultural practices, and drought-related features. A better mechanistic understanding of root-soil-microbiota associations could lead to the development of novel strategies to improve cereal production under drought. In this review, we discuss the root-soil-microbiota interactions for improving the soil environment and host fitness under drought and suggest a roadmap for harnessing the benefits of these interactions for drought-resilient cereals. These methods include conservative trait-based approaches for the selection and breeding of plant genetic resources and manipulation of the soil environments.

Activity of root-knot nematodes associated with composition of a nematode-attached microbiome and the surrounding soil microbiota.

We investigated if activity of the pre-infective juveniles (J2s) of root-knot nematodes is linked to the recruitment of a specific microbiome on the nematode surface and/or to the composition of the surrounding microbiota. For this, we determined the J2 activity (active vs. non-motile, which referred to dead and immobile J2s) upon a 3-day incubation in soil suspensions and studied the composition of bacteria, protists, and fungi present on the nematode surface and in the suspensions using amplicon sequencing of the 16S/18S rRNA genes, and ITS region. We also amended suspensions with Pseudomonas protegens strain CHA0 to study its effects on J2 activity and microbial composition. The J2 activity was suppressed in soil suspensions, but increased when suspensions were amended with P. protegens CHA0. The active and non-motile J2s differed in the composition of surface-attached bacteria, which was altered by the presence of P. protegens CHA0 in the soil suspensions. The bacterial genera Algoriphagus, Pedobacter, and Bdellovibrio were enriched on active J2s and may have protected the J2s against antagonists. The incubation time appeared short for attachment of fungi and protists. Altogether, our study is a step forward in disentangling the complex nematode-microbe interactions in soil for more successful nematode control.

Effects of long-term fertilization with contemporary Danish human urine, composted household waste and sewage sludge on soil nematode abundance and community structure.

It is desirable to recycle the urban waste products human urine, composted household waste and sewage sludge as fertilizers to agricultural fields. This could minimize the use of NPK fertilizer, improve soil structure and store carbon. However, waste products may contain heavy metals, persistent organic pollutants (POP) and plastics, and there are concerns that long-term build-up of these substances will cause unwanted effects on soil health. Nematodes are ubiquitous and numerous in soil ecosystems. Abundance and community structure of soil nematodes can be used as indicators of soil health, as some species are vulnerable to pollution. There are well-developed methods for detecting environmental changes based on nematode community structure. At the long-term CRUCIAL field experiment, where alternative fertilizer products have been applied since 2003, we measured effects of long-term fertilization with human urine, composted household waste and sewage sludge on soil properties (pH, soil organic matter and nitrogen availability), abundance of soil microorganisms (bacteria, fungi, small protozoa and ciliates) and nematode trophic groups compared to plots with unfertilized, NPK and cattle manure treatment. Sampling and assessments were done three times during a growth season. Further, we assessed the composition of nematode communities using metabarcoding. Treatments with a high input of organic matter (cattle manure, composted household waste and sewage sludge) had high abundances of bacteria and thus bacterial grazers (small protozoa, ciliates, and bacterial feeding nematodes). We found a significant correlation between nematode community structure and pH and organic matter. We calculated the nematode Maturity Index 2-5 (pollution indicator) based on metabarcoding data, which did not differ significantly between the treatments. We conclude that long-term fertilization with different types of contemporary Danish urban waste products affects both soil properties and abundance of soil organisms, the latter largely reflecting the organic matter input of the fertilizer treatments. We found no adverse effect on nematode communities that could indicate pollution-induced stress on nematofauna or decreased soil fertility.

Clover Root Uptake of Cereal Benzoxazinoids (BXs) Caused Accumulation of BXs and BX Transformation Products Concurrently with Substantial Increments in Clover Flavonoids and Abscisic Acid.

Metabolomic studies on root uptake and transformation of bioactive compounds, like cereal benzoxazinoids (BXs) in non-BX producing plants, are very limited. Therefore, a targeted mass-spectrometry-based metabolomics study was performed to elucidate the root uptake of BXs in white clover (Trifolium repens L.) and the impact of absorbed BXs on intrinsic clover secondary metabolites. Clover plants grew in a medium containing 100 µM of individual BXs (five aglycone and one glycoside BXs) for 3 weeks. Subsequently, plant tissues were analyzed by liquid chromatography-tandem mass spectrometry to quantify the BXs and clover secondary metabolite concentrations. All BXs were taken up by clover roots and translocated to the shoots. Upon uptake of 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA), 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), 2-hydroxy-1,4-benzoxazin-3-one (HBOA), and 2-ß-d-glucopyranosyloxy-1,4-benzoxazin-3-one (HBOA-glc), the parent compounds and a range of transformation products were seen in the roots and shoots. The individual BX concentrations ranged from not detected (nd) to 469 µg/g of dry weight (dw) and from nd to 170 µg/g of dw in the roots and shoots, respectively. The root uptake of BXs altered the composition of intrinsic clover secondary metabolites. In particular, the concentration of flavonoids and the hormone abscisic acid increased substantially in comparison to control plants.

Distinct chemical resistance-inducing stimuli result in common transcriptional, metabolic, and nematode community signatures in rice root and rhizosphere.

Induced resistance (IR), a phenotypic state induced by an exogenous stimulus and characterized by enhanced resistance to future (a)biotic challenge, is an important component of plant immunity. Numerous IR-inducing stimuli have been described in various plant species, but relatively little is known about 'core' systemic responses shared by these distinct IR stimuli and the effects of IR on plant-associated microbiota. In this study, rice (Oryza sativa) leaves were treated with four distinct IR stimuli (ß-aminobutyric acid, acibenzolar-S-methyl, dehydroascorbic acid, and piperonylic acid) capable of inducing systemic IR against the root-knot nematode Meloidogyne graminicola and evaluated their effect on the root transcriptome and exudome, and root-associated nematode communities. Our results reveal shared transcriptional responses-notably induction of jasmonic acid and phenylpropanoid metabolism-and shared alterations to the exudome that include increased amino acid, benzoate, and fatty acid exudation. In rice plants grown in soil from a rice field, IR stimuli significantly affected the composition of rhizosphere nematode communities 3 d after treatment, but by 14 d after treatment these changes had largely reverted. Notably, IR stimuli did not reduce nematode diversity, which suggests that IR might offer a sustainable option for managing plant-parasitic nematodes.

Genetic disruption of Arabidopsis secondary metabolite synthesis leads to microbiome-mediated modulation of nematode invasion.

In-depth understanding of metabolite-mediated plant-nematode interactions can guide us towards novel nematode management strategies. To improve our understanding of the effects of secondary metabolites on soil nematode communities, we grew Arabidopsis thaliana genetically altered in glucosinolate, camalexin, or flavonoid synthesis pathways, and analyzed their root-associated nematode communities using metabarcoding. To test for any modulating effects of the associated microbiota on the nematode responses, we characterized the bacterial and fungal communities. Finally, as a proxy of microbiome-modulating effects on nematode invasion, we isolated the root-associated microbiomes from the mutants and tested their effect on the ability of the plant parasitic nematode Meloidogyne incognita to penetrate tomato roots. Most mutants had altered relative abundances of several nematode taxa with stronger effects on the plant parasitic Meloidogyne hapla than on other root feeding taxa. This probably reflects that M. hapla invades and remains embedded within root tissues and is thus intimately associated with the host. When transferred to tomato, microbiomes from the flavonoid over-producing pap1-D enhanced M. incognita root-invasion, whereas microbiomes from flavonoid-deficient mutants reduced invasion. This suggests microbiome-mediated effect of flavonoids on Meloidogyne infectivity plausibly mediated by the alteration of the abundances of specific microbial taxa in the transferred microbiomes, although we could not conclusively pinpoint such causative microbial taxa.

Rice diterpenoid phytoalexins are involved in defence against parasitic nematodes and shape rhizosphere nematode communities.

Rice diterpenoid phytoalexins (DPs) are secondary metabolites with a well known role in resistance to foliar pathogens. As DPs are also known to be produced and exuded by rice roots, we hypothesised that they might play an important role in plant-nematode interactions, and particularly in defence against phytoparasitic nematodes. We used transcriptome analysis on rice roots to analyse the effect of infection by the root-knot nematode Meloidogyne graminicola or treatment with resistance-inducing chemical stimuli on DP biosynthesis genes, and assessed the susceptibility of mutant rice lines impaired in DP biosynthesis to M. graminicola. Moreover, we grew these mutants and their wild-type in field soil and used metabarcoding to assess the effect of impairment in DP biosynthesis on rhizosphere and root nematode communities. We show that M. graminicola suppresses DP biosynthesis genes early in its invasion process and, conversely, that resistance-inducing stimuli transiently induce the biosynthesis of DPs. Moreover, we show that loss of DPs increases susceptibility to M. graminicola. Metabarcoding on wild-type and DP-deficient plants grown in field soil reveals that DPs significantly alter the composition of rhizosphere and root nematode communities. Diterpenoid phytoalexins are important players in basal and inducible defence against nematode pathogens of rice and help shape rice-associated nematode communities.

A field study reveals links between hyperaccumulating Sedum plants-associated bacterial communities and Cd/Zn uptake and translocation.

Hyperaccumulating ecotypes of Sedum plants are promising Cd/Zn phytoextractors, with potential for leveraging its rhizospheric or endophytic microbiomes to improve phytoremediation efficiency. However, research of bacteria associated with Sedum at field scale is still lacking. Here, we presented a detailed investigation of the bacterial microbiome of hyperaccumulating Sedum ecotypes (S. alfredii and S. plumbizincicola) and a non-hyperaccumulating S. alfredii ecotype, which grow at different soil environments. Moreover, we evaluated the heavy metal uptake and translocation potential of Sedum plants at different locations. The results showed that both HE ecotypes, contrary to the NHE, were efficient for phytoremediation in mine areas and farmlands. For NHE plants, rhizosphere co-occurrence networks were more complex than the networks of other compartments, while networks of HE plants were more complex in bulk soil and roots. The proportion of positive correlations within co-occurrence networks was higher for the HE plants, suggesting a greater potential for mutualistic interactions. Plant compartment and location predominantly shaped the microbiome assembly, and Proteobacteria, Actinobacteria and Acidobacteria dominated the bacterial communities of Sedum plants. Keystone taxa related to Zn hyperaccumulation are similar to those related to Cd hyperaccumulation, and nine bacterial genera had significantly positive correlation with Cd/Zn hyperaccumulation. Taxa, linked to phytoremediation in both mine and farmland (i.e. Actinospica and Streptomyces from Actinobacteria), could be targets for further investigation of their ability to promote metal phytoremediation of Sedum species.

Can microorganisms assist the survival and parasitism of plant-parasitic nematodes?

Plant-parasitic nematodes (PPNs) remain a hardly treatable problem in many crops worldwide. Low efficacy of many biocontrol agents may be due to negligence of the native microbiota that is naturally associated with nematodes in soil, and which may protect nematodes against microbial antagonists. This phenomenon is more extensively studied for other nematode parasites, so we compiled these studies and drew parallels to the existing knowledge on PPN. We describe how microbial-mediated modulation of host immune responses facilitate nematode parasitism and discuss the role of Caenorhabditis elegans-protective microbiota to get an insight into the microbial protection of PPNs in soil. Molecular mechanisms of PPN-microbial interactions are also discussed. An understanding of microbial-aided PPN performance is thus pivotal for efficient management of PPNs.

Phytohormones selectively affect plant parasitic nematodes associated with Arabidopsis roots.

Phytohormones may affect plant-nematode interactions directly as chemo-attractants or -repellents, or indirectly through the root-associated microbiome or through host defense mechanisms. However, the exact roles of phytohormones in these complex plant-soil-nematode interactions are not well understood. We used Arabidopsis thaliana mutants impaired in phytohormone synthesis or sensitivity to elucidate their role in root-nematode interactions. As root-associated microorganisms may modulate these interactions, we explored correlations between the relative abundances of root-associated nematodes, and bacteria and fungi using amplicon sequencing. We found distinct shifts in relative abundances of a range of nematode taxa in the A. thaliana phytohormone mutants. The root knot nematode Meloidogyne hapla, a sedentary endoparasitic species that is in intimate contact with the host, was highly enriched in JA-, SA- and SL-impaired lines, and in an ET-insensitive line. Positive or negative correlations between specific microbial and nematode taxa were observed, but, as the inference of causal relationships between microbiome responses and effects on nematode communities is premature, this should be studied in detail in future studies. In conclusion, genetic derailment of hormonal balances generally rendered plants vulnerable to endoparasitic nematode attack. Furthermore, preliminary data suggest that this effect may be partially modulated by the associated microbiome.

Benzoxazinoids selectively affect maize root-associated nematode taxa.

Although the effects of plant secondary metabolites on plant defence have been studied for decades, the exact roles of secondary metabolites in shaping plant-associated microbial and nematode communities remain elusive. We evaluated the effects of benzoxazinoids, a group of secondary metabolites present in several cereals, on root-associated nematodes. We employed 18S rRNA metabarcoding to compare maize root-associated nematode communities in a bx1 knockout maize line impaired in benzoxazinoid synthesis and in its parental wild type. Both genotype and plant age affected the composition of the nematode community in the roots, and the effects of benzoxazinoids on nematode communities were stronger in the roots than in the rhizosphere. Differential abundance analysis and quantitative PCR showed that the root lesion nematode Pratylenchus neglectus was enriched in the bx1 mutant line, while another root lesion nematode, Pratylenchus crenatus, was reduced. Correlation analysis showed that benzoxazinoid concentrations in maize roots mostly correlated negatively with the relative abundance of nematode sequence reads. However, positive correlations between benzoxazinoids and nematode taxa, including several plant-parasitic nematodes, were also identified. Our detailed nematode community analysis suggests differential and selective effects of benzoxazinoids on soil nematodes depending on both the nematode species and the benzoxazinoid compound.

The plant-growth promoting bacteria promote cadmium uptake by inducing a hormonal crosstalk and lateral root formation in a hyperaccumulator plant Sedum alfredii.

Plant growth-promoting bacteria (PGPB) that inhabit hyperaccumulating plants assist cadmium (Cd) absorption, but the underlying mechanism has not been comprehensively studied. For this reason, we combined the fluorescence imaging, and transcriptomic and metabolomic methods in a Cd hyperaccumulator, Sedum alfredii, inoculated or not with PGPB Pseudomonas fluorescens. The results showed that the newly emerged lateral roots, that were heavily colonized by P. fluorescens, are the main entry for Cd influx in S. alfredii. Inoculation with P. fluorescens promoted a lateral root formation of its host plant, leading to a higher Cd phytoremediation efficiency. Furthermore, the plant transcriptome revealed that 146 plant hormone related genes were significantly up-regulated by the bacterial inoculation, with 119 of them showing a complex interaction, which suggests that a hormonal crosstalk participated root development. The targeted metabolomics analysis showed that P. fluorescens inoculation significantly increased indole acetic acid concentration and significantly decreased concentrations of abscisic acid, brassinolide, trans-zeatin, ethylene and jasmonic acid in S. alfredii roots, thereby inducing lateral root emergence. Altogether, our results highlight the importance of PGPB-induced lateral root formation for the increased Cd uptake in a hyperaccumulating plant.

Pseudomonas fluorescens promote photosynthesis, carbon fixation and cadmium phytoremediation of hyperaccumulator Sedum alfredii.

Plant growth-promoting bacteria (PGPB) can promote photosynthesis and biomass production of hyperaccumulators, achieving enhanced phytoremediation efficiency of cadmium (Cd). A better understanding of the mechanisms controlling photosynthesis of hyperaccumulating plants by PGPB is necessary for developing strategies that promote the practical phytoextraction of Cd-polluted soils. In this study, chlorophyll fluorescence, gas exchange, and transcriptome sequencing were conducted to evaluate the physiological and transcriptional changes on photosynthesis and carbon fixation in hyperaccumulator Sedum alfredii after inoculation with PGPB Pseudomonas fluorescens. The results showed that bacterial inoculation significantly enhanced maximum quantum yield of PS II (Fv/Fm), effective quantum yield of PS II (ΦPSII), photochemical quenching (qP) and chlorophyll concentration, while reduced non-photochemical quenching (NPQ) of S. alfredii. Further, inoculation resulted in an increased net photosynthetic rates (Pn), intercellular CO2 concentration (Ci), transpiration rate (Tr) and stomatal conductance (Gs) of the studied plant. At the transcriptional level, 70 photosynthetic genes and 42 C4-pathway carbon fixation related genes were significantly up-regulated in response to inoculation, which could be the reason for enhanced photosynthesis and dry biomass. To sum up, this P. fluorescens strain can simultaneously promote growth and Cd uptake of S. alfredii, which can be a promising bacterial agent applied to Cd phytoremediation practices.

AgNO3 Sterilizes Grains of Barley (Hordeum vulgare) without Inhibiting Germination-A Necessary Tool for Plant-Microbiome Research.

To understand and manipulate the interactions between plants and microorganisms, sterile seeds are a necessity. The seed microbiome (inside and surface microorganisms) is unknown for most plant species and seed-borne microorganisms can persist and transfer to the seedling and rhizosphere, thereby obscuring the effects that purposely introduced microorganisms have on plants. This necessitates that these unidentified, seed-borne microorganisms are removed before seeds are used for studies on plant-microbiome interactions. Unfortunately, there is no single, standardized protocol for seed sterilization, hampering progress in experimental plant growth promotion and our study shows that commonly applied sterilization protocols for barley grains using H2O2, NaClO, and AgNO3 yielded insufficient sterilization. We therefore developed a sterilization protocol with AgNO3 by testing several concentrations of AgNO3 and added two additional steps: Soaking the grains in water before the sterilization and rinsing with salt water (1% (w/w) NaCl) after the sterilization. The most efficient sterilization protocol was to soak the grains, sterilize with 10% (w/w) AgNO3, and to rinse with salt water. By following those three steps, 97% of the grains had no culturable, viable microorganism after 21 days based on microscopic inspection. The protocol left small quantities of AgNO3 residue on the grain, maintained germination percentage similar to unsterilized grains, and plant biomass was unaltered. Hence, our protocol using AgNO3 can be used successfully for experiments on plant-microbiome interactions.

Effect of ash application on the decomposer food web and N mineralization in a Norway spruce plantation.

In the face of global climate change there is an increasing demand for biofuel, which exerts pressure on production and thus management of biofuel plantations. The intensification of whole-tree harvest from biofuel plantations increases export of nutrients. Returning ash from biofuel combustion to the forest plantations can amend the soil nutrient status and thus facilitate sustainable forest management. However, ash affects the forest floor decomposer food web, potentially changing organic matter turnover, carbon sequestration and nitrogen availability. Our aim was to examine the response of decomposer organisms, food web structure and nitrogen mineralization function after ash application. In a coniferous forest plantation amended with 0, 3, 4.5 or 6 t ash ha-1, we sampled in several depths of the forest floor for key organisms of the decomposer food web (fungal biomass, 0-12 cm; bacteria, protozoa, nematodes and enchytraeids, 0-3 cm and 3-6 cm; microarthropods and earthworms, 0-5 cm), 2, 14 and 26 months after ash application. We used structural equation modelling (SEM) to detangle the direct and indirect effects of ash application on organisms in the decomposer food web and on nitrogen availability. We found that ash increased the abundance of bacteria and protozoa, as well as the inorganic nitrogen pool at 0-3 cm depth, whereas the effect of ash was negligible at 3-6 cm depth. Earthworm abundance increased, whereas enchytraeid abundance decreased 2 years after ash application. The structural equation modelling showed that ash application stimulated the bacterial feeding pathway and increased nitrogen mineralization. Contrary, ash had a negative effect on fungal biomass at the first sampling, however, this effect subdued over time. Our results suggest that as the soil decomposer food web is resilient to ash application, this is a viable option for sustainable management of biofuel plantations.

Wood ash effects on growth and cadmium uptake in Deschampsia flexuosa (Wavy hair-grass).

Wood ash recycling to forests is beneficial because it regains nutrients and prevents acidification, but wood ash application is restricted due to its cadmium (Cd) content. We question if Cd in wood ash represents a problem, since decreases in Cd bioavailability due to ash-induced pH changes may counteract increased total Cd concentration. We studied effects of wood ash (0, 3, 9 and 30 t ha-1) and lime (pH increase equivalent to the wood ash treatments) on growth and Cd uptake in Deschampsia flexuosa. After four months, we measured plant biomass and Cd accumulation, and extracted Cd from the soil using three different methods; HNO3 (total), EDTA (chelator-based) and NH4NO3 (salt-based). Wood ash and lime strongly stimulated plant growth. Cd concentration in the plant tissue decreased with wood ash and lime addition, and correlated positively with the NH4NO3 extractable fraction of Cd in the soil. In contrast, HNO3 and EDTA extracted more Cd with increased wood ash application. We conclude that wood ash amendment increases soil pH, total Cd concentration, nutrient levels and stimulates plant growth. However, it does not increase Cd accumulation in D. flexuosa, as pH-driven decreases in Cd bioavailability leads to reduced plant Cd uptake. Finally, soil bioavailable Cd is best determined using NH4NO3-extraction.

Wood ash decreases cadmium toxicity to the soil nematode Caenorhabditis elegans.

Wood ash is a beneficial fertilizer and liming agent in nutrient depleted soils, but it also contains considerable amounts of cadmium (Cd), which can be toxic to organisms in the environment. Therefore, risk assessments regarding utilization of wood ash is required. Here, we studied how wood ash (applied in doses equivalent to 0, 3 and 6 t ha-1) and Cd (applied in doses of 0, 10, 150, 300, 600, 1200 and 2000 mg kg-1) affected growth of the soil nematode Caenorhabditis elegans. The treatments were combined in a full factorial design. Wood ash alone greatly stimulated both soil respiration and growth of C. elegans, whereas Cd alone had a toxic effect. However, unrealistically high Cd levels were needed to severely affect growth of C. elegans and soil respiration, especially soil respiration was very resilient to Cd amendment. Ash addition decreased Cd toxicity to C. elegans, with an EC50 value of 390 mg Cd kg-1 in the 3 t ash ha-1 treatment, and an increase of EC50 to 1894 mg Cd kg-1 in the 6 t ash ha-1 treatment. This is probably because ash increases the Cd sorption capacity of the soil, and thereby decreases the bio-availability of Cd. The results suggest that there is no acute toxic effect of Cd to nematodes associated with wood ash recycling; in fact, our results suggest that ash actually decrease Cd toxicity.

Impacts of Root Metabolites on Soil Nematodes.

Plant parasitic nematodes cause significant crop damage globally. Currently, many nematicides have been banned or are being phased out in Europe and other parts of the world because of environmental and human health concerns. Therefore, we need to focus on sustainable and alternative methods of nematode control to protect crops. Plant roots contain and release a wide range of bioactive secondary metabolites, many of which are known defense compounds. Hence, profound understanding of the root mediated interactions between plants and plant parasitic nematodes may contribute to efficient control and management of pest nematodes. In this review, we have compiled literature that documents effects of root metabolites on plant parasitic nematodes. These chemical compounds act as either nematode attractants, repellents, hatching stimulants or inhibitors. We have summarized the few studies that describe how root metabolites regulate the expression of nematode genes. As non-herbivorous nematodes contribute to decomposition, nutrient mineralization, microbial community structuring and control of herbivorous insect larvae, we also review the impact of plant metabolites on these non-target organisms.