Plants are continuously exposed to volatile organic compounds (VOCs) emitted by neighbors. Although the mechanisms governing VOC uptake and metabolism remain unclear, cytochrome P450 monooxygenases (CYP450s) are thought to participate in the detoxification and metabolic conversion of absorbed VOCs. Data from previously conducted cDNA microarray transcriptomic profiling in potato exposed to French marigold essential oil (FM-EO) was here used to filter differentially expressed sequences, and identified 54 unique CYP450 transcripts. Among the 10 most highly expressed sequences, two CYP81D1-like (81D1-1 and 81D1-2) and one CYP81D11-like (81D11-1) transcripts were found. RT-qPCR confirmed their strong induction within 8 h of volatile exposure. Comprehensive bioinformatics identified the most highly induced 81D1-1 gene as a CYP450 containing a predicted N-terminal hydrophobic signal or membrane-anchor region, the conserved heme-binding signature motif, and regulatory elements associated with oxidative stress responses. The other 81D11-1 gene, exhibiting a comparable expression level, was annotated only as a heme-binding protein but possessed seven distinct cis-regulatory elements, suggesting high transcriptional plasticity. Machine learning predictions assigned the highest interaction probability to (Z)-β-ocimene, whereas structure-based docking yielded the most favorable mean score for piperitone. This study provides the first characterization of the potato CYP450 superfamily in the context of volatile-mediated plant–plant interactions and identifies two CYP81D members as strong candidates for the oxidative metabolism of absorbed VOCs. The results support a proposed detoxification pathway in which CYP81-mediated oxidation precedes glutathione conjugation and intracellular sequestration of VOCs. These candidate genes provide a valuable foundation for future functional studies and may facilitate the development of sustainable crop protection strategies based on volatile-mediated plant defense.

β-cyclodextrin (β-CD) is a good host for the encapsulation of fennel and basil essential oils (FEO and BEO, respectively) and the formation of inclusion complexes (ICs) using the co-precipitation method. According to the results of the GC/MS analysis conducted in this study, monoterpenes and monoterpenoids were the dominant chemical groups in total FEO, while in BEO, these two groups occurred along with sesquiterpenes and sesquiterpenoids. The presence of dominant compounds from both EOs was validated using the FT-IR spectra of ICs, which indicated successful complexation. Analyses conducted using SPME/GC-MS showed the continuous emission of volatiles over 24 h from both ICs. Under SEM, particles of both ICs appeared to have a rectangular or rhomboid morphology and few aggregates. The insecticidal properties of EOs and ICs with β-CD were tested on the Colorado potato beetle (CPB) as a model pest. The inclusion complex of β-CD with FEO altered the developmental dynamic and body mass of the CPB. The initial increase in the proteolytic activity of CPB larvae fed with potato plants sprayed with ICs was not maintained for long, and the proteolytic efficacy of treated larvae remained in line with that of the control larvae. Future investigations will focus on manipulating the volume of EOs used and the treatment duration for optimal efficacy and potential application.

Phloretin is a key secondary metabolite produced by apple trees. Known for its strong antioxidant properties, this dihydrochalcone has been extensively studied in animals but less so in plants. Recently, we identified phloretin as a phytotoxic allelochemical that inhibits growth in the model plant Arabidopsis by disrupting auxin metabolism and distribution in the roots. In this study, we found that phloretin significantly hinders the growth of Arabidopsis seedlings' aerial parts after a short-term treatment (10 days) and causes their decay after long-term exposure (28 days). These effects result from ultrastructural damage in the mesophyll cells of the leaves, including chloroplast displacement and swelling, lesions, and alterations in thylakoid and cell wall organization. Interestingly, phloretin-treated plants showed a decrease in malondialdehyde levels and antioxidant enzyme activities, while hydrogen peroxide and proline levels remained unchanged. This suggests that phloretin-induced chlorosis and seedling decay are not due to oxidative stress but rather to severe chloroplast structural damage, leading to inefficient photosynthesis, starch degradation, starvation, and activation of micro- and macroautophagic processes for self-preservation. Ultimately, these processes result in programmed cell death. These new insights into the phytotoxic effects of phloretin on Arabidopsis shoots could pave the way for future research into phloretin as a potential multitarget bioherbicide and enhance our understanding of autoallelopathy in apple trees.

To elucidate the involvement of glutathione in the mitigation of induced oxidative changes and the sequestration of perceived volatiles in cells, we exposed potato plants to French marigold essential oil. The formation of short-lived radicals, the determination of scavenging activity towards ascorbyl and DPPH radicals, and the assessment of the potato plants’ overall intra/extracellular reduction status were performed using electron paramagnetic resonance spectroscopy (EPR). The results showed the presence of hydroxyl radicals in potatoes, with significantly reduced accumulation in exposed plants compared to the control group after 8 h. However, the kinetics of EPR signal intensity change for the pyrrolidine spin probe (3CP) in these plants showed very low reducing potential, suggesting that the antioxidant system acts lethargically and/or the probe has been reoxidized. Total glutathione and its reduced/oxidized form ratio, determined spectrophotometrically, showed that the exposed plants initially had lower glutathione levels with diminutive, reduced form compared to the control. Still, after 8 h, both characteristics were similar to those of the control. RT-qPCR analysis revealed that the volatiles altered the expression of glutathione metabolism-involved genes, especially that of glutathione-S-transferase, after 8 h. Glutathione metabolism was affected by volatiles in the initial response of potato plants exposed to French marigold essential oil, and glutathione molecules were involved in the mitigation of induced oxidative burst.

French marigold ( Tagetes patula L.) is an aromatic plant known for its repellent effects on pests. It is traditionally grown in fields near many vegetable crops, including potato ( Solanum tuberosum L.). However, the effects of marigold essential oil (EO) on the physiology of neighboring crops have been neglected in research. The aim of this study was to establish, for the first time, a comprehensive phytohormone profile of potato plants exposed to French marigold EO for different time periods (4, 8 and 12 h). Endogenous levels of all major phytohormone groups, determined by HPLC–MS analysis, showed altered phytohormone responses of EO-exposed potato plants. The most affected were stress-related phytohormones: abscisic acid–glucose ester, 1-aminocyclopropane-1-carboxylic acid, and jasmonic acid. Increased levels of jasmonic acid, cytokinins, storage form of abscisic acid together with decreased levels of indole-3-acetic acid and ethylene precursor were observed. In most of the analyzed phytohormone groups similar response pattern was observed—an increase in levels after short time exposure (4 h), followed by a decrease to control values after prolonged exposure (8 h and 12 h). Expression levels of genes involved in biosynthesis and catabolism of stress-related phytohormones, obtained by de novo bioinformatic processing of data from cDNA microarray analysis, revealed that jasmonic acid biosynthetic pathway was the most affected, with the highest number of altered transcripts and with one of the genes from this pathway ( 12-oxophytodienoate reductase 1-like ) exhibiting the highest expression rate.

French marigold is an aromatic plant rich in polyphenolic secondary metabolites, which pesticidal potential was examined in this study. Ultra-high-performance liquid chromatography (UHPLC) connected with OrbiTrap mass spectrometer (MS) identified 113 phenolics and revealed the most detailed phytochemistry of French marigold published so far. Depending on plant material (flowers or leaves) and solvents used for extraction (water, methanol, dichloromethane), the phenolic composition varied. Methanol extract of flowers, with 89 identified phenolics and high antioxidant activity statistically comparable with positive control Trolox, was chosen for testing of antifeedant potential against the 3rd and 4th instars of Colorado potato beetle (CPB). A significant reduction in final body mass of 4th larval stage fed with potato leaves coated with methanol extract of flowers in the concentration of 10 mg/mL was observed (157.67 mg vs. 182.26 mg of controls fed with non-treated leaves). This caused delayed molting since treated larvae reached the maximal mass a day after controls and this delay persisted during the entire larval development. Continuous feeding caused a 25% decline in digestive proteolytic activity of the 4th instar in comparison to controls. The results suggest that French marigold methanol extract of flowers could be proposed as a promising antifeedant for CPB management, with an impact on the reduction in the environmental footprint associated with synthetic pesticide application.

Flower strips of French Marigold are commonly used pest repellents in potato fields. However, the effect of French Marigold volatiles on potato metabolism, physiology and induced defense is unknown. Thus, a microarray transcriptome analysis was performed to study the effects of French Marigold essential oil (EO) on laboratory-grown potato. After 8 h of exposure to EO, with gas chromatography/mass spectrometry (GC/MS)-detected terpinolene and limonene as dominant compounds, 2796 transcripts were differentially expressed with fold change >2 compared to expression in controls. A slightly higher number of transcripts had suppressed expression (1493 down- vs. 1303 up-regulated). Since transcripts, annotated to different photosynthesis-related processes, were mostly down-regulated, we selected a set of 10 genes involved in the leaf starch metabolism pathway, and validated microarray patterns using quantitative reverse transcription polymerase chain reaction (RT-qPCR). Except for decreased synthesis and induced decomposition of starch granule in leaves, 8 h long EO exposure slightly elevated the accumulation of sucrose compared to glucose and fructose in subjected potato plants. An in vitro feeding bioassay with Colorado potato beetle showed that EO-induced alternations on transcriptional level and in the sugars' metabolism caused the enhancement of feeding behavior and overall development of the tested larvae. Results of comprehensive analysis of transcriptional responses in potato exposed to French Marigold EO provide a basis for further elucidation of molecular mechanisms underlying eco-physiological interactions in companion planting cropping systems.

Food demands of increasing human population dictate intensification of livestock production, however, environmental stresses could jeopardize producers' efforts. Forage legumes suffer from yield losses and poor nutritional status due to salinity increase of agricultural soils. As tools aimed to reduce negative impacts of biotic or abiotic stresses, proteinase inhibitors (PIs) have been promoted for biotechnological improvements. In order to increase tolerance of Lotus corniculatus L. to salt stress, serine PI, BvSTI, was introduced into this legume using Agrobacterium rhizogenes, with final transformation efficiency of 4.57%. PCR, DNA gel-blot, RT-PCR and in-gel protein activity assays confirmed the presence and activity of BvSTI products in transformed lines. Plants from three selected transgenic lines (21, 73 and 109) showed significant alterations in overall phenotypic appearance, corresponding to differences in BvSTI accumulation. Lines 73 and 109 showed up to 7.3-fold higher number of tillers and massive, up to 5.8-fold heavier roots than in nontransformed controls (NTC). Line 21 was phenotypically similar to NTC, accumulated less BvSTI transcripts and did not exhibit an additional band of recombinant trypsin inhibitor as seen in lines 73 and 109. Exposure of the transgenic lines to NaCl revealed different levels of salt stress susceptibility. The NaCl sensitivity index, based on morphological appearance and chlorophyll concentrations showed that lines 73 and 109 were significantly less affected by salinity than NTC or line 21. High level of BvSTI altered morphology and delayed salt stress related senescence, implicating BvSTI gene as a promising tool for salinity tolerance improvement trials in L. corniculatus.