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

A decade of advances in the study of buckwheat for organic farming and agroecology (2013-2023).

During the last decade, research has shown the environment and human health benefits of growing buckwheat (Fagopyrum spp.). This comprehensive review aims to summarize the major advancements made in the study of buckwheat from 2013 to 2023, focusing on its agronomic characteristics, nutritional value, and potential applications in sustainable agriculture. The review examines the diverse applications of buckwheat in organic and agroecological farming systems, and discusses the ability of buckwheat to control weeds through allelopathy, competition, and other sustainable farming methods, such as crop rotation, intercropping and green manure, while improving soil health and biodiversity. The review also explores the nutritional value of buckwheat. It delves into the composition of buckwheat grains, emphasizing their high protein content, and the presence of essential amino acids and valuable micronutrients, which is linked to health benefits such as lowering cholesterol levels, controlling diabetes and acting against different types of cancer, among others. Finally, the review concludes by highlighting the gaps in current knowledge, and proposing future research directions to further optimize buckwheat production in organic or agroecological farming systems. It emphasizes the need for interdisciplinary collaboration, and the integration of traditional knowledge with modern scientific approaches to unlock the full potential of buckwheat as a sustainable crop.

Specialized Metabolites Accumulation Pattern in Buckwheat Is Strongly Influenced by Accession Choice and Co-Existing Weeds.

Screening suitable allelopathic crops and crop genotypes that are competitive with weeds can be a sustainable weed control strategy to reduce the massive use of herbicides. In this study, three accessions of common buckwheat Fagopyrum esculentum Moench. (Gema, Kora, and Eva) and one of Tartary buckwheat Fagopyrum tataricum Gaertn. (PI481671) were screened against the germination and growth of the herbicide-resistant weeds Lolium rigidum Gaud. and Portulaca oleracea L. The chemical profile of the four buckwheat accessions was characterised in their shoots, roots, and root exudates in order to know more about their ability to sustainably manage weeds and the relation of this ability with the polyphenol accumulation and exudation from buckwheat plants. Our results show that different buckwheat genotypes may have different capacities to produce and exude several types of specialized metabolites, which lead to a wide range of allelopathic and defence functions in the agroecosystem to sustainably manage the growing weeds in their vicinity. The ability of the different buckwheat accessions to suppress weeds was accession-dependent without differences between species, as the common (Eva, Gema, and Kora) and Tartary (PI481671) accessions did not show any species-dependent pattern in their ability to control the germination and growth of the target weeds. Finally, Gema appeared to be the most promising accession to be evaluated in organic farming due to its capacity to sustainably control target weeds while stimulating the root growth of buckwheat plants.

Trans-cinnamaldehyde-related overproduction of benzoic acid and oxidative stress on Arabidopsis thaliana.

Introduction: Trans-cinnamaldehyde is a specialised metabolite that naturally occurs in plants of the Lauraceae family. This study focused on the phytotoxic effects of this compound on the morphology and metabolism of Arabidopsis thaliana seedlings. Material and methods: To evaluate the phytotoxicity of trans-cinnamaldehyde, a dose-response curve was first performed for the root growth process in order to calculate the reference inhibitory concentrations IC50 and IC80 (trans-cinnamaldehyde concentrations inducing a 50% and 80% inhibition, respectively). Subsequently, the structure and ultrastructure of the roots treated with the compound were analysed by light and electron microscopy. Based on these results, the following assays were carried out to in depth study the possible mode of action of the compound: antiauxinic PCIB reversion bioassay, determination of mitochondrial membrane potential, ROS detection, lipid peroxidation content, hormone quantification, in silico studies and gene expression of ALDH enzymes. Results: Trans-cinnamaldehyde IC50 and IC80 values were as low as 46 and 87 µM, reducing the root growth and inducing the occurrence of adventitious roots. At the ultrastructural level, the compound caused alterations to the mitochondria, which were confirmed by detection of the mitochondrial membrane potential. The morphology observed after the treatment (i.e., appearance of adventitious roots) suggested a possible hormonal mismatch at the auxin level, which was confirmed after PCIB bioassay and hormone quantification by GC-MS. The addition of the compound caused an increase in benzoic, salicylic and indoleacetic acid content, which was related to the increased gene expression of the aldehyde dehydrogenase enzymes that can drive the conversion of trans-cinnamaldehyde to cinnamic acid. Also, an increase of ROS was also observed in treated roots. The enzyme-compound interaction was shown to be stable over time by docking and molecular dynamics assays. Discussion: The aldehyde dehydrogenases could drive the conversion of trans-cinnamaldehyde to cinnamic acid, increasing the levels of benzoic, salicylic and indoleacetic acids and causing the oxidative stress symptoms observed in the treated seedlings. This would result into growth and development inhibition of the trans-cinnamaldehyde-treated seedlings and ultimately in their programmed-cell-death.

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.

Testing the role of allelochemicals in different wheat cultivars to sustainably manage weeds.

BACKGROUND: Selecting wheat varieties with allelopathic potential or high competitiveness against weeds is a sustainable solution for organic farming to eliminate the use of synthetic herbicides. Wheat is one of the most economically important crops. This study focuses on screening the allelopathic or competitive potential of four wheat cultivars, Maurizio, NS 40S, Adesso and Element, on two weeds of interest due to acquired herbicide resistance, Portulaca oleracea and Lolium rigidum, through germination and growth bioassays and the identification and quantification of benzoxazinoids (BXZs) and polyphenols (phenolic acids and flavonoids). RESULTS: The different cultivars showed different abilities to manage surrounding weeds and different capacity to exude or accumulate specialized metabolites in the presence of those weeds. Furthermore, each cultivar behaved differently depending on the weed present in the medium. The most efficient cultivar to control the tested monocot and dicot weeds was Maurizio, as it effectively controlled germination and growth of L. rigidum and P. oleracea while exuding large amounts of benzoxazinones through the roots, especially the hydroxamic acids 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one and dihydroxy-2H-1,4-benzoxaxin-3(4H)-one. By contrast, NS 40S, Adesso and Element showed the potential to control the growth of just one of the two weeds through allelopathy or competition. CONCLUSION: This study reveals that Maurizio is the most promising wheat cultivar for sustainable weed control, and that the screening of crop varieties with allelopathic potential, which results in the displacement of synthetic herbicides, is an immediate solution in ecological and sustainable agriculture. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Similarities on the mode of action of the terpenoids citral and farnesene in Arabidopsis seedlings involve interactions with DNA binding proteins.

The sesquiterpene farnesene and the monoterpene citral are phytotoxic natural compounds characterized by a high similarity in macroscopic effects, suggesting an equal or similar mechanism of action when assayed at IC50 concentration. In the present study, a short-time experiment (24 and 48 h) using an imaging spectrofluorometer allowed us to monitor the in-vivo effects of the two molecules, highlighting that both terpenoids were similarly affecting all PSII parameters, even when the effects of citral were quicker in appearing than those of farnesene. The multivariate, univariate, and pathway analyses, carried out on untargeted-metabolomic data, confirmed a clear separation of the plant metabolome in response to the two treatments, whereas similarity in the affected pathways was observed. The main metabolites affected were amino acids and polyamine, which significantly accumulated in response to both treatments. On the contrary, a reduction in sugar content (i.e. glucose and sucrose) was observed. Finally, the in-silico studies demonstrated a similar mechanism of action for both molecules by interacting with DNA binding proteins, although differences concerning the affinity with the proteins with which they could potentially interact were also highlighted. Despite the similarities in macroscopic effects of these two molecules, the metabolomic and in-silico data suggest that both terpenoids share a similar but not equal mechanism of action and that the similar effects observed on the photosynthetic machinery are more imputable to a side effect of molecules-induced oxidative stress.

Short-Term Effects of Trans-Cinnamic Acid on the Metabolism of Zea mays L. Roots.

trans-Cinnamic acid is a phenolic compound widely studied in plant metabolism due to its importance in regulating different plant processes. Previous studies on maize plants showed that this compound could affect plant growth and causes metabolic changes in the leaves when applied. However, its effects on root metabolism are not well known. This study analyses the short-term effect of trans-cinnamic acid on the morphology of vascular bundle elements and metabolism in maize roots. At short times (between 6 and 12 h), there is a reduction in the content of many amino acids which may be associated with the altered nitrogen uptake observed in earlier work. In addition, the compound caused an alteration of the vascular bundles at 48 h and seemed to have changed the metabolism in roots to favor lignin and galactose synthesis. The results obtained complement those previously carried out on maize plants, demonstrating that in the short term trans-cinnamic acid can trigger stress-coping processes in the treated plants.

Ecophysiological Responses of Tall Wheatgrass Germplasm to Drought and Salinity.

Tall wheatgrass (Thinopyrum ponticum (Podp.) Barkworth and D.R. Dewey) is an important, highly salt-tolerant C3 forage grass. The objective of this work was to learn about the ecophysiological responses of accessions from different environmental origins under drought and salinity conditions, to provide information for selecting superior germplasm under combined stress in tall wheatgrass. Four accessions (P3, P4, P5, P9) were irrigated using combinations of three salinity levels (0, 0.1, 0.3 M NaCl) and three drought levels (100%, 50%, 30% water capacity) over 90 days in a greenhouse. The control treatment showed the highest total biomass, but water-use efficiency (WUE), δ13C, proline, N concentration, leaf length, and tiller density were higher under moderate drought or/and salinity stress than under control conditions. In tall wheatgrass, K+ functions as an osmoregulator under drought, attenuated by salinity, and Na+ and Cl- function as osmoregulators under salinity and drought, while proline is an osmoprotector under both stresses. P3 and P9, from environments with mild/moderate stress, prioritized reproductive development, with high evapotranspiration and the lowest WUE and δ13C values. P4 and P5, from more stressful environments, prioritized vegetative development through tillering, showing the lowest evapotranspiration, the highest δ13C values, and different mechanisms for limiting transpiration. The δ13C value, leaf biomass, tiller density, and leaf length had high broad-sense heritability (H2), while the Na+/K+ ratio had medium H2. In conclusion, the combined use of the δ13C value, Na+/K+ ratio, and canopy structural variables can help identify accessions that are well-adapted to drought and salinity, also considering the desirable plant characteristics. Tall wheatgrass stress tolerance could be used to expand forage production under a changing climate.

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

Harmaline 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.

Weed pressure determines the chemical profile of wheat (Triticum aestivum L.) and its allelochemicals potential.

BACKGROUND: Common purslane (Portulaca oleracea) and annual ryegrass (Lolium rigidum) are important infesting weeds of field crops. Herbicides are mostly used for weed suppression, while their environmental toxicity and resistance in weeds against them demand considering alternative options, such as the use of allelopathic crops for weed management. Wheat is an important allelopathic crop and present research focused on the identification and quantification of benzoxazinoids (BXZs) and polyphenols (phenolic acids and flavonoids) of the wheat accession 'Ursita' and to screen its allelopathic impact on P. oleracea and Lolium rigidum through equal-compartment-agar (ECA) method. RESULTS: Weed germination, radicle length, biomass and photosynthetic pigments were altered following co-growth of weeds with Ursita for 10-day. Root exudates from Ursita reduced (29-60%) the seedling growth and photosynthetic pigments of Lolium rigidum depending on co-culture conditions of planting density. Weed pressure caused significant increase in the production of phenolic acids (vanillic, ferulic, syringic and p-coumaric acids) and root exudation of BXZs, in particular benzoxazolin-2-one (BOA), 2-hydroxy-7-methoxy-1,4-benzoxazin-3-one (HMBOA), 2-hydroxy-1,4-benzoxazin3-one (HBOA) and 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA) in wheat tissues (shoots, roots) and exudate in root rhizosphere agar medium in response to co-cultivation with Lolium rigidum and P. oleracea, depending on weed/crop density. CONCLUSION: The work revealed that Ursita is allelopathic in nature and can be used in breeding programs to enhance its allelopathic activity. Meanwhile, there are opportunities to explore allelopathic effect of wheat cultivars to control P. oleracea and Lolium rigidum under field conditions. © 2022 Society of Chemical Industry.

Unraveling Sorghum Allelopathy in Agriculture: Concepts and Implications.

Allelopathy is an ecological phenomenon that involves the production and release of biomolecules from different crops, cultivated plants, and bacteria or fungi into the soil rhizosphere and impacts other organisms in the vicinity. Sorghum possesses vital allelopathic characteristics due to which it produces and releases different biomolecules from its root hairs, stems, and grains. Several studies have reported that sorghum acts as an allelopathic crop, decreasing the growth and eco-physiological attributes of surrounding plants and weeds growing simultaneously or subsequently in the field. Sorghum allelopathy has been exploited in the context of green manure, crop rotations, cover crops, and intercropping or mulching, whereas plant aqueous extracts or powder might be an alternate method of weed control. A diverse group of allelochemicals, including benzoic acid, p-hydroxybenzoic acid, vanillic acid, ferulic acid, chlorogenic acid, m-coumaric acid, p-coumaric acid, gallic acid, caffeic acid, p-hydroxibenzaldehyde, dhurrin, sorgoleone, m-hydroxybenzoic acid and protocatechuic acid, have been isolated and identified from different plant tissues of sorghum and root exudates. These allelochemicals, especially sorgoleone, have been investigated in terms of their mode(s) of action, specific activity and selectivity, release in the rhizosphere and uptake and translocation in sensitive species. The present review describes the importance of sorghum allelopathy as an ecological tool in managing weeds, highlighting the most recent advances in the allelochemicals present in sorghum, their modes of action, and their fate in the ecosystem. Further research should focus on the evaluation and selection of sorghum cultivars with high allelopathic potential, so that sorghum allelopathy can be better utilized for weed control and yield enhancement.

Phytotoxic Effects and Mechanism of Action of Essential Oils and Terpenoids.

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.

Phytotoxic Activity of the Natural Compound Norharmane on Crops, Weeds and Model Plants.

Norharmane is a secondary metabolite that appears in different species of land plants. In this paper, we investigated for the first time the specificity of norharmane through germination and growth tests on some crops as Zeamays L. (maize), Triticumaestivum L. (wheat), Oryza sativa L. (rice) and Lactucasativa L. (lettuce) and weeds as Amaranthusretroflexus L. (amaranth), Echinochloacrus-galli L. (barnyard grass), Plantago lanceolata L. (ribwort), Portulaca oleracea L. (common purslane) and Avenafatua L. (wild oat), and its phytotoxic capacity on the metabolism of adult Arabidopsis thaliana L. (thale cress) by measuring chlorophyll a fluorescence, pigment content, total proteins, osmotic potential and morphological analysis. Norharmane had an inhibitory effect on the germination of A.fatua and P.lanceolata, and the growth of P.oleracea, E.crus-galli and A.retroflexus. On adult A. thaliana plants, the compound was more effective to watering, leading to water stress that compromised the growth of the plants and ultimately affected the photosynthetic apparatus. Therefore, this research shows that norharmane not only affects seedlings' metabolism, but also damages the metabolism of adult plants and can be a potential model for a future bioherbicide given its specificity.

Herbicidal Activity of Thymbra capitata (L.) Cav. Essential Oil.

The bioherbicidal potential of Thymbra capitata (L.) Cav. essential oil (EO) and its main compound carvacrol was investigated. In in vitro assays, the EO blocked the germination and seedling growth of Erigeron canadensis L., Sonchus oleraceus (L.) L., and Chenopodium album L. at 0.125 µL/mL, of Setaria verticillata (L.) P.Beauv., Avena fatua L., and Solanum nigrum L. at 0.5 µL/mL, of Amaranthus retroflexus L. at 1 µL/mL and of Portulaca oleracea L., and Echinochloa crus-galli (L.) P.Beauv. at 2 µL/mL. Under greenhouse conditions, T. capitata EO was tested towards the emergent weeds from a soil seedbank in pre and post emergence, showing strong herbicidal potential in both assays at 4 µL/mL. In addition, T. capitata EO, applied by spraying, was tested against P. oleracea, A. fatua and E. crus-galli. The species showed different sensibility to the EO, being E. crus-galli the most resistant. Experiments were performed against A. fatua testing T. capitata EO and carvacrol applied by spraying or by irrigation. It was verified that the EO was more active at the same doses in monocotyledons applied by irrigation and in dicotyledons applied by spraying. Carvacrol effects on Arabidopsis root morphology were also studied.

A natural indole alkaloid, norharmane, affects PIN expression patterns and compromises root growth in Arabidopsis thaliana.

Norharmane is an indole alkaloid that can be found in several terrestrial plants, as well as in some dinoflagellates and cyanobacteria. The aim of this study was to focus on the way this metabolite impacts the plant metabolism of the model species Arabidopsis thaliana. This metabolite caused increase of secondary and adventitious roots, as well as torsion, toxic effects, and a decrease in root length. Moreover, norharmane altered the cellular arrangement, resulting in unfinished cell walls, decreased auxin content and inhibited PIN proteins activity. All the alterations suggest that norharmane alters polar auxin transport by inhibiting PIN2, PIN3 and PIN7 transport proteins, thus causing a significant inhibitory effect on the growth of A. thaliana seedlings.

Imaging of Chlorophyll a Fluorescence in Natural Compound-Induced Stress Detection.

Imaging of chlorophyll a fluorescence (CFI) represents an easy, precise, fast and non-invasive technique that can be successfully used for discriminating plant response to phytotoxic stress with reproducible results and without damaging the plants. The spatio-temporal analyses of the fluorescence images can give information about damage evolution, secondary effects and plant defense response. In the last years, some studies about plant natural compounds-induced phytotoxicity have introduced imaging techniques to measure fluorescence, although the analysis of the image as a whole is often missed. In this paper we, therefore, evaluated the advantages of monitoring fluorescence images, presenting the physiological interpretation of different possible combinations of the most relevant parameters linked to fluorescence emission and the images obtained.

Transcriptome and binding data indicate that citral inhibits single strand DNA-binding proteins.

The mechanism of phytotoxicity of citral was probed in Arabidopsis thaliana using RNA-Seq and in silico binding analyses. Inhibition of growth by 50% by citral downregulated transcription of 9156 and 5541 genes in roots and shoots, respectively, after 1 h. Only 56 and 62 genes in roots and shoots, respectively, were upregulated. In the shoots, the downregulation increased at 3 h (6239 genes downregulated, vs 66 upregulated). Of all genes affected in roots at 1 h (time of greatest effect), 7.69% of affected genes were for nucleic acid binding functions. Genes for single strand DNA binding proteins (SSBP) WHY1, WHY 2 and WHY3 were strongly downregulated in the shoot up until 12 h after citral exposure. Effects were strong in the root at just 1 h after the treatment and then at 12 and 24 h. Similar effects occurred with the transcription factors MYC-2, ANAC and SCR-SHR, which were also significantly downregulated for the first hour of treatment, and downregulation occurred again after 12 and 24 h treatment. Downregulation of ANAC in the first hour of treatment was significantly (P < 0.0001) decreased more than eight times compared to the control. In silico molecular docking analysis suggests binding of citral isomers to the SSBPs WHY1, WHY2, and WHY3, as well as with other transcription factors such as MYC-2, ANAC and SCR-SHR. Such effects could account for the profound and unusual effects of citral on downregulation of gene transcription.

Transcriptome responses to the natural phytotoxin t-chalcone in Arabidopsis thaliana L.

BACKGROUND: New modes of action are needed for herbicides. The flavonoid synthesis intermediate t-chalcone causes apoptosis-like symptoms in roots and bleaching of shoots of Arabidospsis, suggesting a unique mode of action as a phytotoxin. RESULTS: Using RNA-Seq, transcriptome changes were monitored in Arabidopsis seedlings during the first 24 h of exposure (at 1, 3, 6, 12 and 24 h) to 21 µm t-chalcone (I50 dose), examining effects on roots and shoots separately. Expression of 892 and 1000 genes was affected in roots and shoots, respectively. According to biological classification, many of the affected genes were transcription factors and genes associated with oxidative stress, heat shock proteins, xenobiotic detoxification, ABA and auxin biosynthesis, and primary metabolic processess. These are secondary effects found with most phytotoxins. Potent phytotoxins usually act by inhibiting enzymes of primary metabolism. KEGG pathway analysis of transcriptome results from the first 3 h of t-chalcone exposure indicated several potential primary metabolism target sites for t-chalcone. Of these, p-hydroxyphenylpyruvate dioxygenase (HPPD) and tyrosine amino transferase were consistent with the bleaching effect of the phytotoxin. Supplementation studies with Lemna paucicostata and Arabidiopsis supported HPPD as the target, although in vitro enzyme inhibition was not found. CONCLUSIONS: t-Chalcone is possibly a protoxin that is converted to a HPPD inhibitor in vivo. © 2019 Society of Chemical Industry.

Rosmarinic acid induces programmed cell death in Arabidopsis seedlings through reactive oxygen species and mitochondrial dysfunction.

Phytotoxic potential of rosmarinic acid (RA), a caffeic acid ester largely found in aromatic species, was evaluated on Arabidopsis through metabolomic and microscopic approaches. In-vitro bioassays pointed out that RA affected root growth and morphology, causing ROS burst, ROS scavengers activity inhibition and consequently, an alteration on cells organization and ultrastructure. In particular, RA-treatment (175 µM) caused strong vacuolization, alteration of mitochondria structure and function and a consistent ROS-induced reduction of their transmembrane potential (ΔΨm). These data suggested a cell energy deficit also confirmed by the metabolomic analysis, which highlighted a strong alteration of both TCA cycle and amino acids metabolism. Moreover, the increase in H2O2 and O2- contents suggested that RA-treated meristems underwent oxidative stress, resulting in apoptotic bodies and necrotic cells. Taken together, these results suggest that RA inhibits two of the main ROS scavengers causing high ROS accumulation, responsible of the alterations on mitochondrial ultrastructure and activity through ΔΨm dissipation, TCA-cycle alteration, cell starvation and consequently cell death on Arabidopsis seedlings. All these effects resulted in a strong inhibition on root growth and development, which convert RA in a promising molecule to be explored for further use in weed management.