Tomás Cavaco

The management of plant-parasitic nematodes (PPNs) still relies on traditional nematicides that threaten the environment and human health. Novel solutions are urgently needed for PPN pest management that are effective while safeguarding non-target organisms. Volatile phytochemicals belong to a structurally diverse group of bioactive metabolites that are believed to hold safer environmental characteristics than synthetic pesticides. Nonetheless, not many studies have analysed the potential environmental benefits of shifting to these novel bionematicides. In the present study, 20 phytochemical volatiles with reported nematicidal activity were compared to traditional pesticides using specific parameters of environmental and human health safety available on applied online databases and predicted in silico through specialised software. Overall, the reviewed nematicidal phytochemicals were reportedly less toxic than synthetic nematicides. They were predicted to disperse to the air and soil environmental compartments and were reported to have a lower toxicity on aquatic organisms. On the contrary, the synthetic nematicides were reportedly toxic to aquatic organisms while showing a predicted high affinity to the water environmental compartment. As alternatives, β-keto or fatty acid derivatives, e.g., aliphatic alcohols or ketones, showed more adequate properties. This study highlights the importance of complementing studies on nematicidal activity with a risk assessment-based analysis to allow for a faster selection of nematicidal phytochemical volatiles and to leverage the development and implementation of bionematicides.

Modern farming is heavily dependent on the frequent application of pesticides that are known to disturb plant, animal, and microbial biodiversity (as well as accumulate in the environment and influence human health). Plant parasitic nematodes are some of the most damaging pathogens to food crops, impacting crop yield twice as much as phytophagous insects. To combat these pests, large amounts of pesticides are added to agricultural soils and eventually leach out to adjacent environments, accumulate in food and feed, and become very toxic. Volatiles produced by plants and microbes have been increasingly analysed for their anti-nematodal activities since they can present very active chemical structures suitable for developing novel biopesticides. The present study reviews the benefits of some of the most active compounds found in the literature in comparison to the most widely used commercial nematicides. By resorting to data retrieved from freely available online databases on their experimental properties, and given the use of certified software for the prediction of their environmental, toxicological, and ecotoxicological impacts, namely the US EPA Estimation Program Interface (EPI) and the Toxicity Estimation Software Tool (TEST), the advantages and disadvantages of using volatiles in the development of bionematicides are discussed. Ultimately, shifting to a more sustainable usage of pesticides and to farming practices that favor biodiversity can be beneficial to crop production.

Plant-parasitic nematodes (PPNs) are highly damaging pests responsible for heavy losses in worldwide productivity in a significant number of important plant crops. Common pest management strategies rely on the use of synthetic chemical nematicides, which have led to serious concerns regarding their impacts on human health and the environment. Plant natural products, or phytochemicals, can provide a good source of agents for sustainable control of PPNs, due to their intrinsic characteristics such as higher biodegradability, generally low toxicity for mammals, and lower bioaccumulation in the environment. In this work, the nematicidal activity of 39 phytochemicals was determined against the root-lesion nematode (RLN) Pratylenchus penetrans using standard direct and indirect contact methodologies. Overall, the RLN was tolerant to the tested phytochemicals at the highest concentration, 2 mg/mL, seldom reaching full mortality. However, high activities were obtained for benzaldehyde, carvacrol, 3-octanol, and thymol, in comparison to other phytochemicals or the synthetic nematicide oxamyl. These phytochemicals were seen to damage nematode internal tissues but not its cuticle shape. Also, the environmental and (eco)toxicological parameters reported for these compounds suggest lower toxicity and higher safety of use than oxamyl. These compounds appear to be good candidates for the development of biopesticides for a more sustainable pest management strategy.

This study presents the first report of a Fusarium wilt in Cymbopogon citratus in Portugal. Fusarium is one of the most prominent genera of plant pathogens due to its wide range of hosts and mycotoxin production. The Fusarium fujikuroi species complex (FFSC) encompasses several threatening known plant pathogens. Cymbopogon citratus is a broadly distributed aromatic and medicinal plant rich in bioactive volatiles, which are relevant to several industries. The primary goal of this study was to identify and characterize the Fusarium fujikuroi species putatively responsible for the observed wilt. This report displays symptomatologic, cultural, morphologic, genetic, and cytogenomic characteristics associated with this fungus and disease. The cultural features included flat, white-colored colonies with filiform margins. Additionally, these colonies displayed abundant cottony aerial mycelia at the upper surface and orange-violet color at the lower surface. On Carnation Leaf-Piece Agar, septate fusoid macroconidia were present, displaying a flattened tapering toward the basal part and a number of septa ranging from one to four. The comparison between the amplified and aligned ITS sequences revealed 100% similarity between the isolate and the FFSC. Finally, a flow cytometry assay revealed an estimated genome size of 29.9 Mbp. This finding contrasts with other known pathogens from the Fusarium fujikuroi species complex. Ultimately, novel pathogens might be uncovered by exploring the mycobiome of diseased Cymbopogon citratus plants.

Managing soil fertility is vital for agriculture. However, modern farming excessively relies on mineral fertilizers, which lessens profit and endangers ecosystem health. Grasslands made up of Poaceae and Fabaceae, including woody species, offer feed for livestock, lowers farmers’ economic risks, and conserve resources. Grassland crops can enhance soil fertility in a more sustainable way than mineral fertilization. To counter fertilizer-driven soil decline, permanent grasslands or crop rotations are effective. Also, grassland soils generally contain more nitrogen, potassium and organic matter and less phosphorus than cropland soils. They additionally enhance soil’s physical and biological parameters, limiting erosion while elevating biodiversity. This work focuses on the benefits of grasslands towards crop production, reviewing their influence on soil fertility parameters that boost soil health.

Climate change is prompting a shift of tropical pests to locations with a previously lower probability of invasion. This is the case for root-knot nematodes, Meloidogyne sp., particularly of the tropical group. Among them, M. ethiopica is now considered a threat to European food security. The development of novel sustainable nematicides can be based on in vitro bioassays of highly active phytochemicals, e.g., volatiles from essential oils. However, a steady supply of nematodes is often very difficult and dependent on environmental conditions. In the present study, an in vitro co-culture system of M. ethiopica parasitizing hairy roots of Solanum lycopersicum was established, for the first time, to easily obtain populations of second-stage juveniles (J2). These were then used to screen the nematicidal activity of 10 volatile compounds characteristic of essential oils. Finally, information on the most successful compounds was reviewed to predict their environmental dispersion and ecotoxicological hazards. The M. ethiopica population obtained from the co-culture was morphologically similar to reported populations in natural conditions and could be accurately used in direct-contact bioassays. The aldehydes citral and citronellal induced complete mortality of the tested J2, at 1 mg/mL, while compounds from other chemical groups were not as successful. In comparison to commonly used commercial nematicides, citral and citronellal were less likely to accumulate in the water environmental compartment and have lower reported toxicities compared to aquatic organisms and to mammals. Overall, in vitro co-cultures showed the potential to expedite the screening and discovery of bioactive compounds as a contribution to the development of sustainable biopesticides, as well as to lower the impacts of modern farming on agroecosystems.

Control of the pinewood nematode (PWN), the causal agent of pine wilt disease, can be achieved through the trunk injection of nematicides; however, many pesticides have been linked to environmental and human health concerns. Essential oils (EOs) are suitable alternatives due to their biodegradability and low toxicity to mammals. These complex mixtures of plant volatiles often display multiple biological activities and synergistic interactions between their compounds. The present work profiled the toxicity of eight EOs against the PWN in comparison to their 1:1 mixtures, to screen for successful synergistic interactions. Additionally, the main compounds of the most synergistic mixtures were characterized for their predicted environmental fate and toxicity to mammals in comparison to emamectin benzoate, a commercial nematicide used against PWN. The mixtures of Cymbopogon citratus with Mentha piperita and of Foeniculum vulgare with Satureja montana EOs showed the highest activities, with half-maximal effective concentrations (EC₅₀) of 0.09 and 0.05 µL/mL, respectively. For these, complete PWN mortality was reached after only ca. 15 min or 2 h of direct contact, respectively. Their major compounds had a higher predicted affinity to air and water environmental compartments and are reported to have very low toxicity to mammals, with low acute oral and dermal toxicities. In comparison, emamectin benzoate showed lower nematicidal activity, a higher affinity to the soil and sediments environmental compartments and higher reported oral and dermal toxicity to mammals. Overall, uncovering synergistic activities in combinations of EOs from plants of different families may prove to be a source of biopesticides with optimized toxicity against PWNs.

The pinewood nematode (PWN), Bursaphelenchus xylophilus, the causal agent of pine-wilt disease (PWD), is a threat to Pinus forests in Asia and Europe. Bursaphelenchus mucronatus and B. fraudulentus are closely related non-pathogenic pine-wood nematodes. In the present work, four medium-chain aliphatic alcohols (C10 to C13) were evaluated in direct-contact bioassays against B. xylophilus, B. mucronatus and B. fraudulentus. The compounds showed high nematicidal activity against the species tested. The lowest values for half-maximal effective concentrations (EC₅₀) were determined in B. xylophilus and B. fraudulentus, suggesting a higher sensibility to these compounds. Further bioassays will include compounds with different chain lengths and functional groups to explore the diversity in the activity of oxygen-containing aliphatic compounds for a more targeted sustainable-control strategy for the PWN.

The pinewood nematode (PWN) Bursaphelenchus xylophilus is a major threat to pine forests. Research on sustainable pest management strategies is now a priority. Essential oils (EOs) are biodegradable, complex mixtures of volatiles that can show low toxicity to mammals and powerful nematicidal activities depending on their chemical composition. In the present work, the EOs of four plants were screened against the PWN, solely and in combination to identify possible synergistic interactions. The combination of Cymbopogon citratus and Mentha piperita EOs (1:1) resulted in higher activities than those of each tested solely, suggesting the occurrence of synergistic interactions between the compounds of these EOs. Research on the combination of synergistic EOs may lead to the development of plant based biopesticides with optimized activities against the PWN.