Phytotoxic Effects and Mechanism of Action of Essential Oils and Terpenoids

[EN] Weeds are one of the major constraints in crop production affecting both yield and quality. The excessive and exclusive use of synthetic herbicides for their management is increasing the development of herbicide-resistant weeds and is provoking risks for the environment and human health. Therefore, the development of new herbicides with multitarget-site activity, new modes of action and low impact on the environment and health are badly needed. The study of plant-plant interactions through the release of secondary metabolites could be a starting point for the identification of new molecules with herbicidal activity. Essential oils (EOs) and their components, mainly terpenoids, as pure natural compounds or in mixtures, because of their structural diversity and strong phytotoxic activity, could be good candidates for the development of new bioherbicides or could serve as a basis for the development of new natural-like low impact synthetic herbicides. EOs and terpenoids have been largely studied for their phytotoxicity and several evidences on their modes of action have been highlighted in the last decades through the use of integrated approaches. The review is focused on the knowledge concerning the phytotoxicity of these molecules, their putative target, as well as their potential mode of action.This research was supported by the Ministerio de Ciencia, Innovacion y Universidades RTI2018-094716-B-100 and by the Italian Ministry of Education, University and Research (MIUR), project SIR-2014 cod. RBSI14L9CE (MEDANAT).Verdeguer Sancho, MM.; Sánchez-Moreiras, AM.; Araniti, F. (2020). Phytotoxic Effects and Mechanism of Action of Essential Oils and Terpenoids. Plants. 9(11):1-52. https://doi.org/10.3390/plants9111571S15291

The influence of environment on invasive Carpobrotus sp. populations across genetic clusters

The study aims to explore the natural variation in the metabolome of different populations of the invasive plant Carpobrotus from different genetic clusters and geographical origins to enhance our comprehension of its involvement in the adaptation process and phenotypic diversity. The metabolomic profile of shoots was analysed in four populations from two different genetic clusters (Cluster A: Cádiz and A Lanzada; Cluster B: La Marina and Samil) and two different biogeographical regions in Spain (Atlantic: Samil and A Lanzada; Mediterranean: Cádiz and La Marina), collected in the field and subsequently grown in the greenhouse. In addition, climatic, and physiological parameters were analysed. The Mediterranean populations (Cádiz and La Marina) showed lower initial weight and length measurements in morphological parameters than the Atlantic populations. On the contrary, only root parameters showed significant differences in growth parameters among populations. The analysis of ion levels revealed a consistent pattern of higher concentrations in shoots compared to roots, with significant differences among populations, particularly in sodium (Na+) and chlorides (Cl−) levels. Regarding metabolomic analysis, clear correlations between the metabolome, genetic and climatic conditions of Carpobrotus sp.pl populations are described. Pairwise comparisons using t-tests and Principal Component Analysis (PCA) indicated that the differences in metabolomic profile between the Samil and La Marina populations, which correspond to the same genetic cluster (cluster B), were smaller than in the rest of the comparisons indicating that populations from the same genetic cluster were more similar metabolically than those from the same climatic region. The study identified key metabolites representative of each cluster, with significant differences in amino acids, organic acids, and sugars contributing to the variation among populations. Pathway analysis highlighted the impact of climatic conditions on metabolic pathways, particularly in populations from Cluster A. In conclusion, the different populations were more similar according to the genetic cluster than to the climatic region of origin when studied at the metabolomic level. Consequently, the metabolites more representative of each cluster were also identified.Universidade de Vigo/CISU

Application of indole-alkaloid harmaline induces physical damage to photosystem II antenna complexes in adult plants of Arabidopsis thaliana (L.) Heynh

Finding herbicides with new and multiple modes of action is a solution to stop the increase in resistant weed species. Harmaline, a natural alkaloid with proven phytotoxic potential, was tested on Arabidopsis adult plants by watering and spraying; watering resulted as the more effective treatment. Harmaline altered several photosynthetic parameters, reducing the efficiency of the light- (ΦII) and dark-adapted (Fv/Fm) PSII, suggesting physical damages in photosystem II, although dissipation of the energy in excess under the form of heat was not compromised as demonstrated by the significant increase in ΦNPQ. Metabolomic alterations, such as osmoprotectant accumulation and reduction in sugars’ content, also indicate a reduction of photosynthetic efficiency and suggest early senescence and water status alteration induced by harmaline. Data suggest that harmaline might be considered a new phytotoxic molecule interesting for further studies.Ministerio de Ciencia e Innovación | Ref. RTI2018-094716-B-100Universidade de Vigo/CISU

Azelaic acid can efficiently compete for the auxin binding site TIR1, altering auxin polar transport, gravitropic response, and root growth and architecture in Arabidopsis thaliana roots

The present study investigates the phytotoxic potential of azelaic acid (AZA) on Arabidopsis thaliana roots. Effects on root morphology, anatomy, auxin content and transport, gravitropic response and molecular docking were analysed. AZA inhibited root growth, stimulated lateral and adventitious roots, and altered the root apical meristem by reducing meristem cell number, length and width. The treatment also slowed down the roots' gravitropic response, likely due to a reduction in statoliths, starch-rich organelles involved in gravity perception. In addition, auxin content, transport and distribution, together with PIN proteins' expression and localisation were altered after AZA treatment, inducing a reduction in auxin transport and its distribution into the meristematic zone. Computational simulations showed that AZA has a high affinity for the auxin receptor TIR1, competing with auxin for the binding site. The AZA binding with TIR1 could interfere with the normal functioning of the TIR1/AFB complex, disrupting the ubiquitin E3 ligase complex and leading to alterations in the response of the plant, which could perceive AZA as an exogenous auxin. Our results suggest that AZA mode of action could involve the modulation of auxin-related processes in Arabidopsis roots. Understanding such mechanisms could lead to find environmentally friendly alternatives to synthetic herbicides.Agencia Estatal de Investigación | Ref. RT12018-094716-B-I00Xunta de Galicia | Ref. ED481A-2021/32

Ultrastructural and hormonal changes related to harmaline-induced treatment in Arabidopsis thaliana (L.) Heynh. root meristem

Financiado para publicación en acceso aberto: Universidade de Vigo/CISUGHarmaline is an indole alkaloid with demonstrated phytotoxicity and recognized pharmacological applications. However, no information is available concerning its mode of action on plant metabolism. Therefore, the present work evaluated bioherbicide mode of action of harmaline on plant metabolism of Arabidopsis thaliana (L.) Heynh. Harmaline induced a strong inhibitory activity on root growth of treated seedlings, reaching IC50 and IC80 values of 14 and 29 μM, respectively. Treated roots were shorter and thicker than control and were characterized by a shorter root meristem size and an increase of root hairs production. Harmaline induced ultrastructural changes such as increment of cell wall thickness, higher density and condensation of mitochondria and vacuolization, appearance of cell wall deposits, increment of Golgi secretory activity and higher percentage of aberrant nuclei. The ethylene inhibitor AgNO3 reversed high root hair appearance and increment of root thickness, and pTCSn::GFP transgenic line showed fluorescence cytokinin signal in stele zone after harmaline treatment that was absent in control, whereas the auxin signal in the transgenic line DR5 was significantly reduced by the treatment. All these results suggest that the mode of action of harmaline could be involving auxin, ethylene and cytokinin synergic/antagonistic action.Ministerio de Ciencia, Innovación y Universidades | Ref. RTI2018-094716-B-10

Potential Effects of Essential Oils Extracted from Mediterranean Aromatic Plants on Target Weeds and Soil Microorganisms

[EN] Essential oils (EOs), extracted from aromatic plants, have been proposed as candidates to develop natural herbicides. This study aimed to evaluate the herbicidal potential of Thymbra capitata (L.) Cav., Mentha x piperita L. and Santolina chamaecyparissus L. essential oils (EOs) on Avena fatua L., Echinochloa crus-galli (L.) P. Beauv, Portulaca oleracea L. and Amaranthus retroflexus L. and their effects on soil microorganisms. A pot experiment was set up and three EOs at three doses were applied by irrigation. Efficacy and effects of EOs on weed growth were determined. Soil microbial biomass carbon and nitrogen, microbial respiration, and the main microbial groups were determined at days 7, 28 and 56. EOs demonstrated herbicidal activity, increasing their toxicity with the dose. T. capitata was the most effective against all weeds at the maximum dose. P. oleracea was the most resistant weed. Soil microorganisms, after a transient upheaval period induced by the addition of EOs, recovered their initial function and biomass. T. capitata EO at the highest dose did not allow soil microorganisms to recover their initial functionality. EOs exhibited great potential as natural herbicides but the optimum dose of application must be identified to control weeds and not negatively affect soil microorganisms.Jouini, A.; Verdeguer Sancho, MM.; Pinton, S.; Araniti, F.; Palazzolo, E.; Badalucco, L.; Laudicina, VA. (2020). Potential Effects of Essential Oils Extracted from Mediterranean Aromatic Plants on Target Weeds and Soil Microorganisms. Plants. 9(10):1-24. https://doi.org/10.3390/plants9101289S12491

Control of Problematic Weeds in Mediterranean Vineyards with the Bioherbicide Pelargonic Acid

[EN] Pelargonic acid (PA) is the only natural herbicide authorized for professional use in Spain. Incorporating PA into an integrated weed management strategy in vineyards may enable a more sustainable production method for grapes. In this work, PA of 55% concentration, formulated by a commercial company (PSEI), was evaluated and applied at 8, 10, 12, and 15 L/ha for weed control in Mediterranean vineyards during 2020 and 2021. A total of 22 different weed species, 16 dicotyledonous and 6 monocotyledonous, were identified in the experimental areas. Previously, greenhouse assays were performed against Avena fatua L. and Chenopodium album L. to determine the dose/response curves. PSEI proved to be a viable post-emergence herbicide with an efficacy of 40.79¿80.90%, depending on the applied dose (higher doses were the most effective). Broader herbicidal activity (20% or more) was obtained against dicotyledonous weeds compared with monocotyledonous. The PA formulation was remarkable in achieving PSEI-similar effects as compared to the market reference but at lower concentrations (around 13% less PA) and doses (1¿8 less L/ha). PA has proved to be a good candidate to control weeds in Mediterranean vineyards when used as a post-emergence broad-spectrum herbicide in the first stages of weed development.This research was funded by SEIPASA.Muñoz, M.; Torres-Pagán, N.; Jouini, A.; Araniti, F.; Sánchez-Moreiras, AM.; Verdeguer Sancho, MM. (2022). Control of Problematic Weeds in Mediterranean Vineyards with the Bioherbicide Pelargonic Acid. Agronomy. 12(10):1-18. https://doi.org/10.3390/agronomy12102476118121

Differential olive grove management regulates the levels of primary metabolites in xylem sap

Aims The conventional management adopted in many Mediterranean olive orchards makes them more vulnerable to climate change and attacks by pathogens, due to the decreased chemical plant defenses. In this scenario, a metabolomic analysis was carried out on the xylem sap (Xsap) of olive plants (Olea europaea L.) grown in the Salento peninsula (Italy). Methods Trials were carried out in two olive groves, one organically and one conventionally managed (controls), successively both converted to sustainable management (i.e. frequent light pruning, soil and foliar fertilization, cover crops). The Xsap was extracted from the shoots of olive plants using a Scholander pressure chamber pressurized with N2 and gas chromatography-mass spectrometry metabolite profiling was performed in the Xsap. Results An untargeted gas chromatography mass spectrometry (GC-MS) based metabolomic analysis of primary metabolites (including underivatized volatiles) of the Xsap revealed relative abundances of 153 identified metabolites and 336 unknown features across the 12 samples from four groups of samples. Among them, more than half were involved in the primary metabolism. Many of the compounds with increased levels under sustainable management (such as amino acids, soluble sugars, sugar alcohols) have a well-known role as osmoprotectants or are involved in plant defense, growth and development during stress or recovery stages. Conclusions Sustainable management in olive groves can increase the ability of plants to overcome environmental stressors and enhance ecosystem balance

Investigating the phytotoxic potential of Carlina acaulis essential oil against the weed Bidens pilosa through a physiological and metabolomic approach

Essential oils (EOs) are widely studied as possible candidates for new eco-friendly herbicides for weed management due to their phytotoxicity. In this study we tested the phytotoxic potential of the EO obtained from the roots of Carlina acaulis L. (Apiaceae) against the weed Bidens pilosa L. This EO, containing 98% of the polyacetylene carlina oxide, showed strong phytotoxic effects on the plant metabolism, such as leaf necrosis, reduction of relative water content and total leaf area, and an increase in the dry weight/fresh weight ratio, suggesting a water status alteration. The EO also damaged the photosynthetic machinery, as evidenced by the significant reduction of the effective quantum yield of photosystem II (ΦII) and the maximum quantum yield of photosystem II (Fv/Fm). In addition, the non-photochemical quenching (ΦNPQ) significantly increased after spraying with C. acaulis EO. Damage to photosystem II was further demonstrated through the reduction of manganese and calcium concentrations, possibly due to an alteration in the correct functionality of the Mn4Ca cluster of the PSII. Metabolomics analysis revealed an accumulation of branched-chain amino acids, such as isoleucine and valine, which is commonly related to osmotic alterations under drought stress situations and a general reduction in sugar content (fructose, glucose, mannose, among others), suggesting reduction of the photosynthetic efficiency too. Overall, these findings suggest C. acaulis EO as a promising natural product with phytotoxic potential against weeds that deserves further investigation

Trichoderma spp.-mediated mitigation of heat, stress, and their combination on the Arabidopsis thaliana holobiont: a metabolomics and metabarcoding approach

IntroductionThe use of substances to increase productivity and resource use efficiency is now essential to face the challenge of feeding the rising global population with the less environmental impact on the ecosystems. Trichoderma-based products have been used as biopesticides, to inhibit pathogenic microorganisms, and as biostimulants for crop growth, nutrient uptake promotion, and resistance to abiotic stresses.MethodsIn this work, plant metabolomics combined with roots and rhizosphere bacterial metabarcoding were exploited to inspect the performance of Trichoderma spp. biostimulants on Arabidopsis thaliana under drought, heat and their combination and its impact on plant holobiont.Results and discussionAn overall modulation of N-containing compounds, phenylpropanoids, terpenes and hormones could be pointed out by metabolomics. Moreover, metabarcoding outlined an impact on alpha and beta-diversity with an abundance of Proteobacteria, Pseudomonadales, Burkholderiales, Enterobacteriales and Azospirillales. A holobiont approach was applied as an integrated analytical strategy to resolve the coordinated and complex dynamic interactions between the plant and its rhizosphere bacteria using Arabidopsis thaliana as a model host species