Investigation on Chemical Composition, Antioxidant, Antifungal and Herbicidal Activities of Volatile Constituents from Deverra tortuosa (Desf.)

This study aims to analyze the chemical composition of the essential oils (EOs) obtained from stems and umbels of D. tortuosa as well the assessment of their biological activity. EOs were extracted by hydrodistillation and analyzed by gas chromatography coupled to mass spectrometry (GC/MS). The antioxidant properties were determined by DPPH and ABTS assays. The phytotoxic potential was assessed against dicots weeds (Sinapis arvensis and Trifolium campestre), monocots weeds (Lolium rigidum) and the crop Lepidium sativum. The antifungal activity was evaluated against four target phytopathogenic fungal strains. High diversity of compounds was detected in D. tortuosa Eos, varying among plant parts and consisting mainly of α-pinene (24.47–28.56%), sabinene (16.2–18.6%), α-phellandrene (6.3–11.7%) and cis-ocimene (5.28–7.85%). D. tortuosa EOs exhibited remarkable antioxidant activity, as well as interesting variable antifungal activities depending on the dose and fungi strain. The herbicidal activity of EOs showed significant efficacy on the inhibition of germination and seedling growth of all tested herbs. These results suggest that the EOs of Deverra tortuosa represent a valuable source of antioxidant, antifungal and phytotoxic metabolites and could be potential candidates for pest management, contributing to the promotion of sustainable agriculture.


Introduction
Appreciation and utilization of natural bioactive substances in aromatic plants have increased in recent years [1]. It is known that aromatic plants are considered as alternatives for microbial resistance and as a useful avenue for safe pest control and management [1,2]. Currently, pest-related problems are largely controlled through the application of chemical pesticides [3]. This generalized use makes it possible to control the majority of pests, but it induces generally undesirable side effects whose acuity is increasing, such as influence on the microflora of the soil, development of biotypes resistant to pesticides, contamination of the environment and generation of harmful effects on human health [4]. In addition to the changes that affect our environment and climate, they also affect the whole dynamic of life by disturbing the ecosystems and the species that constitute them. These effects have an impact, in particular, on the interactions between species and their environment [5,6]. These interactions are crucial for the maintenance of ecosystems where chemical mediation plays from a flame ionization detector (FID); b = calculated linear retention index relative to a series of n-alkanes (C8-C30) on a HP-5MS column; c = linear retention index relative to the literature data [38][39][40][41]; d = identification method; RI: retention index; MS = mass spectrometry; Inj = co-injection with authentic compounds; compounds higher than 5.0% are highlighted in boldface; -= absent.
A clear variability of the chemical composition between the EOs from stems and umbels was observed, both in the presence/absence and in the amounts of the components ( Figure 1). A clear variability of the chemical composition between the EOs from stems and umbels was observed, both in the presence/absence and in the amounts of the components ( Figure 1). Both EOs were characterized by a richness in hydrocarbon monoterpenes with a rate exceeding 80% of the total. The main components ( Figure 2) of this fraction were α-pinene (24.2-28.8%), sabinene (16.23-18.67%), β-pinene (5.1-6.2%) and cis-ocimene (5.28-7.85%) for umbels and stems, respectively, with similar amounts for stems and umbels; however, α-phellandrene varied between 6.3 and 11.74% in umbels and stems, respectively. for umbels and stems, respectively, with similar amounts for stems and umbels; however, α-phellandrene varied between 6.3 and 11.74% in umbels and stems, respectively. However, β-phellandrene (6.7%) was only detected in the EO from stems, and p-cymene (6.77%) was detected only in umbels. p-cymene is a monoterpene, naturally present in certain plants, in particular, in Apiaceae species. This compound is known to have several biological activities [42,43] and is an important industrial intermediate used in the synthesis of fungicides, pesticides, perfumes and fragrances, and also in the production of some precursors of standard antioxidants such as p-cresol.
Oxygenated sesquiterpenes were represented by three compounds, with a total ranging between 4.17 and 5.61% for umbels and stems, respectively. The major component of this fraction was spathulenol, which varied between 2.56 and 3.74%.
Three phenylpropanoid derivatives, apiole, methyl eugenol and elemicin, were detected in low amounts ( Figure 3). Apiole, a characteristic metabolite of the family Apiaceae, was detected only in umbels EOs. Finally, two phthalide derivatives were detected. Their chemical structures are represented in Figure 4. However, β-phellandrene (6.7%) was only detected in the EO from stems, and pcymene (6.77%) was detected only in umbels. p-cymene is a monoterpene, naturally present in certain plants, in particular, in Apiaceae species. This compound is known to have several biological activities [42,43] and is an important industrial intermediate used in the synthesis of fungicides, pesticides, perfumes and fragrances, and also in the production of some precursors of standard antioxidants such as p-cresol.
Oxygenated sesquiterpenes were represented by three compounds, with a total ranging between 4.17 and 5.61% for umbels and stems, respectively. The major component of this fraction was spathulenol, which varied between 2.56 and 3.74%.
Three phenylpropanoid derivatives, apiole, methyl eugenol and elemicin, were detected in low amounts ( Figure 3). Apiole, a characteristic metabolite of the family Apiaceae, was detected only in umbels EOs. However, β-phellandrene (6.7%) was only detected in the EO from stems, and p-cymene (6.77%) was detected only in umbels. p-cymene is a monoterpene, naturally present in certain plants, in particular, in Apiaceae species. This compound is known to have several biological activities [42,43] and is an important industrial intermediate used in the synthesis of fungicides, pesticides, perfumes and fragrances, and also in the production of some precursors of standard antioxidants such as p-cresol.
Oxygenated sesquiterpenes were represented by three compounds, with a total ranging between 4.17 and 5.61% for umbels and stems, respectively. The major component of this fraction was spathulenol, which varied between 2.56 and 3.74%.
Three phenylpropanoid derivatives, apiole, methyl eugenol and elemicin, were detected in low amounts ( Figure 3). Apiole, a characteristic metabolite of the family Apiaceae, was detected only in umbels EOs. Finally, two phthalide derivatives were detected. Their chemical structures are represented in Figure 4. Finally, two phthalide derivatives were detected. Their chemical structures are represented in Figure 4.
The composition reported proves the variability in the EOs from stems and umbels. Phthalide have been found mainly in plants from the Apiaceae family, such as Apium graveolens L., Angelica sinensis (Oliv.) Diels, A. acutiloba (Siebold & Zucc.) Kitag., Levisticum officinale W.D.J. Koch, Cnidium officinale Makino and Ligusticum porteri J.M.Coult. & Rose [43][44][45][46]. The presence of this group of compounds confers to the Apiaceae family several biological activities, as phthalides are a class of secondary metabolites with a wide range of pharmacological and agronomical activities.   [43][44][45][46]. The presence of this group of compounds confers to the Apiaceae family several biological activities, as phthalides are a class of secondary metabolites with a wide range of pharmacological and agronomical activities.
It is known that the production of EOs and secondary metabolites varies depending on the pedoclimatic conditions in the first instance and also on the collection and extraction conditions. The variability of production according to pedoclimatic conditions and the origin of the plant material is called ecological chemistry. The latter is a source of biodiversity and a richness in active molecules synthesized by plants following interactions with their biotopes. Ecological chemistry contributes significantly to the discovery and bioinspiration for the search for active molecules. In the case of D. tortuosa, a great variability in compounds and chemotypes has been described in the literature [21,22,[34][35][36][37]47]. Table 2 reports the literature data on the chemical variability of D. tortuosa EOs.   It is known that the production of EOs and secondary metabolites varies depending on the pedoclimatic conditions in the first instance and also on the collection and extraction conditions. The variability of production according to pedoclimatic conditions and the origin of the plant material is called ecological chemistry. The latter is a source of biodiversity and a richness in active molecules synthesized by plants following interactions with their biotopes. Ecological chemistry contributes significantly to the discovery and bioinspiration for the search for active molecules. In the case of D. tortuosa, a great variability in compounds and chemotypes has been described in the literature [21,22,[34][35][36][37]47]. Table 2 reports the literature data on the chemical variability of D. tortuosa EOs.     According to the literature, the chemical composition of D. tortuosa EOs has been described for plants growing in Tunisia, Libya, Egypt and Algeria. Different chemotypes have been described, and the EOs varied depending on the area, season, plant part, and condition (fresh/dry) of the plant material [21,22,[34][35][36][37]47]. The variability in yield and chemical composition reflects the interaction and the close relationship between the production of molecules and their ecological environment.

Antioxidant Activity
The antioxidant potential of the EOs was evaluated by two free radical scavenging methods. The first method is the DPPH test since it makes it possible to measure the antiradical power of pure molecules or plant extracts in a model system. It measures the ability of an antioxidant to reduce the chemical radical DPPH (2,2-diphenyl-1-picrylhydrazyl) by hydrogen transfer. The EOs were evaluated with reference to the antioxidant potential of gallic acid. The results are expressed as IC 50 (µL of EO/mL of DPPH solution). The results obtained are shown in Figure 5. Both EOs showed a potential for trapping DPPH with IC50 values of 66.21 and mg/mL for stem and umbel, respectively. On the other hand, this activity was mod by comparing it with the activity of gallic acid.
The radical scavenging activity was also evaluated by the ABTS (2,2′-azinoethylbenzothiazolin-6-sulfonic acid) cation radical decolorization test. The main are shown in Figure 6. Both EOs showed a potential for trapping DPPH with IC 50 values of 66.21 and 55.87 mg/mL for stem and umbel, respectively. On the other hand, this activity was moderated by comparing it with the activity of gallic acid.
The radical scavenging activity was also evaluated by the ABTS (2,2 -azino-bis-(3ethylbenzothiazolin-6-sulfonic acid) cation radical decolorization test. The main results are shown in Figure 6. Figure 5. IC50 of the EOs expressed as IC50 (mg/mL). Means followed by the same letter are not significantly different according to multivariate analysis ANOVA (p ≤ 0.05). Data are the mean of three replicates, and ± bars indicate the standard deviation of the mean.
Both EOs showed a potential for trapping DPPH with IC50 values of 66.21 and 55.87 mg/mL for stem and umbel, respectively. On the other hand, this activity was moderated by comparing it with the activity of gallic acid.
The radical scavenging activity was also evaluated by the ABTS (2,2′-azino-bis-(3ethylbenzothiazolin-6-sulfonic acid) cation radical decolorization test. The main results are shown in Figure 6.  Both oils showed antioxidant potential for scavenging ABTS radicals, with IC 50 values of 32.36 and 39.57 mg/mL for stem and umbel EOs, respectively. These activities were significant but moderate in comparison with ascorbic acid (IC 50 = 5.35 mg/mL).
The Apiaceae family is rich in antioxidant metabolites. The antioxidant activities of EOs and crude extracts have been described in the literature for Foeniculum vulgare Miller [11], Anegelica acutiloba [48], Daucus carota L. [49], and even D. tortuosa [35]. On the other hand, the antioxidant potential observed in this study can be related to the chemical composition, which showed a richness in monoterpenes, sesquiterpenes, some phenylpropanoids and phthalides. All these compounds are known for their antioxidant activities [50]. According to the literature, the antioxidant potential of monoterpenes, including the main constituents of D tortuosa EOs, such as α-pinene, sabinene and cisocimene, has been reported [51]. Apiole, elemicin and methyl eugenol have also been reported for their antioxidant activities as well as phthalide derivatives [48].

Phytotoxic Activity
In the current study, D. tortuosa EOs have been tested for their herbicidal potential against the germination and growth of weed dicots (S. arvensis and T. campestre) and monocots (L. rigidum) and also against the crop, L. sativum (Figure 7), and phytotoxicity was compared to the synthetic herbicide, glyphosate. The results are shown in Tables 3-5. reported [51]. Apiole, elemicin and methyl eugenol have also been reported for their antioxidant activities as well as phthalide derivatives [48].

Phytotoxic Activity
In the current study, D. tortuosa EOs have been tested for their herbicidal potential against the germination and growth of weed dicots (S. arvensis and T. campestre) and monocots (L. rigidum) and also against the crop, L. sativum (Figure 7), and phytotoxicity was compared to the synthetic herbicide, glyphosate. The results are shown in Tables 3-5.   0 ± 0 Ca 0 ± 0 Ca 0 ± 0 Ca 3 0 ± 0 Ca 0 ± 0 Ca 0 ± 0 Ca 4 0 ± 0 Ca 0 ± 0 Ca 0 ± 0 Ca Data are the mean of three replicates ± SD. Within each species, different capital letters in the same column indicate that means are different among concentrations, and different lowercase letters in the same line compare the activity of EOs and glyphosate for the same species and the same dose according to multivariate analysis ANOVA (p ≤ 0.05).  The EOs showed significant effects on the germination and growth of the aerial and root parts of the tested plants.
Both EOs significantly inhibited the germination of all tested seeds in a dose-dependent manner. At 1 mg/mL, the germination of T. campestre was around 100%, and no significant effects on the inhibition of germination either by D. tortuosa EOs or glyphosate were detected. T. campestre was more sensitive to stem EO with a total inhibition at 3 mg/mL. At the same dose, the inhibition was 33% with umbel EO and 43% with glyphosate.
A total inhibition of germination of L. sativum was obtained at 3 mg/mL for both EOs, whereas glyphosate determined a partial inhibition (33%).
S. arvensis and L. rigidum were the most sensitive to the action of the Eos. At 2 mg/mL, germination was totally inhibited, as with glyphosate.
These results show a significant anti-germinative activity of the Eos, which often exceeds that of glyphosate. The inhibition results varied depending on the tested seed and also the EO.
The EOs were phytotoxic not only in the inhibition of germination but also in the growth of the aerial parts and roots when compared to the control. The EOs inhibited the growth of shoots of tested species with a dose-response effect ( Table 4). Both EOs, at 1 mg/mL, inhibited by 50% the growth of shoots of T. campestre, which germination was not affected by the same dose. S. arvensis, also resistant to germination inhibition, resulted in total inhibition at 1 mg/mL. At the same dose, the shoot length of L. rigidum was inhibited by 72.8 and 77.1% by the EOs of stems and umbels, respectively. At 2 mg/mL, the shoot length of L. sativum was severely inhibited by 61.5 and 72.1%, respectively, by stem and umbel EOs. The inhibitory potency of the EOs on shoot growth was similar to that of glyphosate.
The inhibitory activity of both EOs was also significant on radical growth (Table 5). At 2 mg/mL, L. sativum root length was inhibited by 72.8 and 88.5%, respectively, by the EOs of stems and umbels, and the radical length of T. campestre showed inhibition of 88.5 and 89.5%, respectively. The stem and umbel EOs, at 1 mg/mL, provoked inhibition of root leghth of L. rigidum by 62.7 and 73.1%, respectively.
It has been proven that even if some seeds of the test species were able to germinate, they were successively inhibited in both shoot and root length in a combined phytotoxicity event.
These results agree with the literature. In fact, Eos and aqueous and organic extracts (petroleum ether, acetone and chloroform) from the aerial parts of D. tortuosa were evaluated for their allelopathic potential against two cultivated species, Brassica rapa L. and Linum usitatissimum L., showing a remarkable herbicidal potential [32]. The EOs contain high amounts of monoterpene hydrocarbons (81.05 and 81.84 for stems and umbels, respectively). This class of terpenes is known for its herbicidal potential and its involvement in the allelopathic potential of several plants. The results obtained in this study prove the close relationship between the richness in monoterpene hydrocarbons in D. tortuosa EOs, neglecting the possible interactions of synergism and antagonism of the different components detected in the two EOs [52][53][54][55][56].
Sabinene, present in appreciable amounts in the EOs (16.23-18.67%), has been reported for its herbicidal properties, with inhibition of the growth of shoots and roots of Poa annua L. and Amaranthus retroflexus [57]. Moreover, the EO of a sabinene-rich chemotype of Ravensara aromatica Sonn. was reported to have herbicidal properties against the germination and growth of garden cress (Lepidium sativum L.) and rice (Oryza sativa L.) [58].
Several studies showed that the involvement of EOs and/or their pure components in the phenomenon of allelopathy is linked to the induction of oxidative stress, leading to the alteration of the vital functions of plants [56]. Furthermore, some EOs have been shown to inhibit germination and growth following spray application, provoking an alteration of membrane integrity via the peroxidation of fatty acids from membrane phospholipids, resulting in the release of malondialdehyde, blocking chlorophyll synthesis, inducing proline accumulation and resulting in plant desiccation [53].
α-and β-pinene, in appreciable amounts in the EOs reported in this research, have been reported to induce physiological effects resulting in the inhibition of germination and growth of several weeds [54][55][56]. Indeed, the application of these two compounds on seeds of Cassia occidentalis L. and Oryza sativa induced an inhibition of chlorophyll synthesis and an increase in the content of macromolecules such as proteins and carbohydrates, accompanied by an inhibition of activity of some hydrolyzing enzymes such as proteases, α-amylases and β-amylases. On the other hand, the activity of peroxidases, superoxide dismutase, guaiacol peroxidase, catalase, ascorbate reductase and polyphenol oxidases increased, reflecting the oxidative stress generated by the application of these molecules [54,56]. Similarly, pinenes have been reported to disrupt energy metabolism on roots of maize seedling by inhibiting oxidative phosphorylation and the electron transport chain [50]. All of these results prove the importance of essential oils of D. tortuosa as candidate molecules for allelopathic applications in sustainable agriculture.

Antifungal Activity of D. tortuosa EOs
EOs exhibiting interesting antifungal activities depending on the tested strains, EOs were screened at different concentrations. Statistical analysis revealed a significant difference concerning the concentration of each oil and also significant differences among the inhibitory effects of the EOs. Similarly, strains of phytopathogens reacted in different ways to the action of the EOs (Figure 8). The results are shown in Table 6.
Plants 2023, 12, x FOR PEER REVIEW 13 of 19 [50]. All of these results prove the importance of essential oils of D. tortuosa as candidate molecules for allelopathic applications in sustainable agriculture.

Antifungal Activity of D. tortuosa EOs
EOs exhibiting interesting antifungal activities depending on the tested strains, EOs were screened at different concentrations. Statistical analysis revealed a significant difference concerning the concentration of each oil and also significant differences among the inhibitory effects of the EOs. Similarly, strains of phytopathogens reacted in different ways to the action of the EOs (Figure 8). The results are shown in Table 6.   The umbel EOs showed the most significant effects compared to the stems. A total inhibition was obtained at the dose of 8 mg/mL against F. oxysporum solani and at a dose of 10 mg/mL for the other strains tested, and these two doses were retained as MIC. However, the inhibition by the stem EO, even at 10 mg/mL, was always partial. F. lycopersici and F. oxysporum mathioli were the two most resistant strains to the action of stem EO.
The results obtained confirm the antimicrobial potential of D. tortuosa EOs, which have been reported on bacteria and other strains of phytopathogenic fungi. In fact, Krifa et al. [32] reported a total inhibition of the growth of Fusarium graminearum and Alternaria sp. at 6 mg/mL of D. tortuosa EO. Sabinene, one of the major components of D. tortuosa EOs, has been described as having bactericidal activity, and its inhibitory effects on biofilm formation and pH tolerance have been reported [59].
Phthalide derivatives, detected in our study in the EOs, are known for their antifungal potential. These molecules, in particular 3-n-butylidenephthalide, are known to have antifungal potential that often exceeds that of synthetic fungicides such as itraconazole, fluconazole and ketoconazole [60][61][62]. The presence of these compounds could contribute to the activity of D. tortuosa Eos and also explains the difference in activity between umbel and stem EOs.
The antifungal activity of EOs can be attributed to the effect of secondary metabolites synthesized by plants in the context of their allelopathic interactions with the different components of their biotopes. This antimicrobial effect depends on the substances with antifungal properties present in each plant and each biotope.

Plant Material
Aerial parts of Deverra tortuosa (stems and umbels) were harvested from the Djerissa region located in the northwest of Tunisia, characterized by a semi-arid bioclimatic stage. Five different samples from different plants separated by at least 50 m were collected and then dried in a glass greenhouse for 10 days; then, the plant material was used for the extraction of the EOs. For the herbicidal assays, mature seeds of Trifolium campestre Schreb, Sinapis arvensis L., Lepidium sativum L. and Lolium rigidum Gaudin were collected from crop fields ( Table 7). Voucher specimens of collected plants (Table 7) were identified by Professor Hamrouni Lamia and deposited at the Laboratory of Genetic and Forest Ecology of the National Research Institute of Rural Engineering, Water and Forests, Tunisia.

Extraction of the Essential Oils
The EOs were extracted using a Clevenger-type apparatus for four hours. The EOs were weighed for yield calculation and then dried over anhydrous sodium sulfate and stored in sealed glass bottles at 4 • C. The yield was calculated based on the dry weight of samples by the following formula: Oil yield (%, W/W) = weight of essential oils (g) dried weight of plant material (g) * 100

Gas Chromatography Analysis with MS Detection
The GC-MS analysis of the EOs was carried out on an Agilent 6890 gas chromatograph equipped with a fused silica capillary column HP-5MS (30 m × 0.25 mm film, 5% phenylmethyl-siloxane and 0.25 µm). The EOs were dissolved in n-hexane (2 mg/mL). The injection of 1 µL of each sample was carried out in split mode with a ratio of 1:100; helium was used as the carrier gas at a rate of 1 mL/min. The operative conditions were as follows: temperature of injector 250 • C, temperature of detector 280 • C. The oven temperature varied as follows: 35 • C to 325 • C with a rate of 5 • C/min. The equilibration time was 0.5 min. MS operate in electron impact was at a potential of 70 eV in a scan range from 50-550 m/z. The identification of the components was based on their Kovats retention indices and their comparison with those of the literature [38][39][40][41], and their mass spectra with those available in the NIST 02 and Wiley 275 mass spectral libraries [63]. The Kovats indices were calculated related to a series of n-alkanes (C8-C30). This identification of some components was confirmed by co-injection with authentic compounds: α-pinene, sabinene, α-phellandrene, α-terpinolene, methyl eugenol and β-phellandrene (purchased from Sigma-Aldrich, Germany). Quantification was electronically recorded from FID area data using ChromCard program, Thermoquest.

2,2 -Diphenyl-1-picrylhydrazyl (DPPH) Assay
The free radical scavenging activity was assessed using the DPPH assay, according to the method described by Ud-Daula et al. (2016) [64]. A total of 300 µL of samples, EOs or butylated hydroxytoluene (purchased from Sigma-Aldrich, Germany) used as standard at different concentrations or methanol (blank), were added to 2 mL of freshly prepared DPPH (purchased from Sigma-Aldrich, Darmstadt, Germany) methanolic solution (0.1 mM). The mixture was vortexed and incubated at room temperature for 30 min in the dark, and the absorbance was then measured at 517 nm against a blank on a Biowave IIWPA spectrophotometer (Serlabo Technologies). The antiradical activity was expressed as IC 50 (mg/mL), the amount of sample required to scavenge 50% of free radicals present in the test solution. All experiments were conducted in triplicate, and data are expressed as mean values ± SD

ABTS •+ Free Radical Scavenging Activity
The ABTS •+ radical assay was performed according to procedures reported by Ud-Daula et al. [64]. A solution containing ABTS •+ radical cations (ABTS + ) was prepared by mixing an equal volume of potassium persulfate (2.45 mM) and ABTS (7 mM) (purchased from Sigma-Aldrich, Darmstadt, Germany). The mixture was then incubated for 16 h at room temperature in the dark. The final obtained solution was then diluted with ethanol to an absorbance of 0.70 at 734 nm. A total of 200 µL of various concentration of tested EOs (5-100 mg/mL) and ascorbic acid (used as a standard) were mixed with 2 mL of the ABTS radical solution and allowed to incubate in the dark at room temperature for 5 min. Then, the absorbance was measured at 734 nm on a Biowave IIWPA spectrophotometer (Serlabo Technologies). The inhibition percentage was calculated against the blank. All experiments were conducted in triplicate; results are expressed as IC 50, the concentration of tested sample needed to scavenge 50% of ABTS radical cation.

Herbicidal Activity of Deverra tortuosa Eos
Seeds of Sinapis arvensis, Trifolium campestre, Lepidium sativum and Lolium rigidum were used in phytotoxic activity assays. Before germination tests, seeds were disinfected with 5% sodium hypochlorite. Ten seeds were placed in Petri dishes lined with double-layer filter paper Whatman No.1 and treated with different doses (1, 2, 3, 4 mg/mL) of the EOs in a solution of Tween 20 (0.1%) [65]. The herbicidal potential of the EOs was compared to the activity of the synthetic herbicide, glyphosate. Phytotoxic assays were performed in a completely randomized design, and each assay was replicated three times. After 10 days, the germination percentages were calculated, and the growth of roots and shoots was measured in cm.

Fungal Strains
Four phytopathogenic fungal strains were used. F. oxysporum solani was obtained from the laboratory of plant protection of the Tunisian National Institute of Agronomic Research, and Fusarium oxysporum sp. lycopersici, F. oxysporum mathioli and F. culmorum were obtained from Dr Hirsch's Laboratory, University of California Los Angeles (UCLA).

In Vitro Antifungal Activities on Mycelial Growth
The antifungal properties of the EOs were tested in vitro in Petri dishes using the agar dilution method [66]. The samples were diluted in a Tween 20 solution (0.1% v/v) and then added to 10 mL of PDA at 40 • C to provide the required concentrations (6, 8 and 10 µL/mL). PDA plates containing only Tween 20 (0.1%) were used as the negative control. A mycelial disk of 6 mm in diameter was placed in each PDA plate, then incubated at 24 • C for 7 days. All assays were repeated with three replicates for each strain and each dose. The fungicidal properties were determined as the percentage of inhibition (PI) of mycelia growth compared to the control, following the formula: where dc and dt are the mean diameter of control growth and treated fungi, respectively. Minimum inhibitory concentration is determined as the lowest dose at which there is no fungal growth.

Statistical Analysis
EOs yield, antioxidants tests, germination and seedling growth experiments and antifungal assays were carried out using a randomized block design and three replications for each assay. Statistical analyses were performed with Statistical Package for the Social Sciences (SPSS version 23.0. IBM Corp.: Armonk, NY, USA). Results were examined statistically using one-way analysis of variance (ANOVA) followed by Student-Newman-Keuls tests. The differences between individual means were considered significant at p ≤ 0.05.

Conclusions
Plants always contain a mixture of several biological molecules that act simultanously or differently, take similar or different paths, and act together or independently on one or more targets, thus leading to an important biological potential, contributing to an organic and producible agriculture, healthy environment, and also a rich and sustainable ecosystem. In this sense, the EOs of Deverra tortuosa showed a richness and diversity of compounds. These oils vary depending on the part of the plant (umbels or stems), the same with the literature that is related to the origin of the plant, the season of collection and the drying of the plant material. This makes this species a source of various metabolites with various interests, above all, that in this current study, several biological activities have been demonstrated. Indeed, the antioxidant, antimicrobial and also herbicidal potential have been proven during this study, which may suggest a possible use of D. toruosa EOs in sustainable agricultural practices, both as herbicides and antifungals. Furthermore, these EOs can form the basis of environmentally friendly formulations.