Elsevier Editorial System(tm) for Industrial Crops and Products Manuscript Draft Manuscript Number: INDCRO-D-17-01815R3 Title: Growth, photosynthetic pigments, phenolic content and biological activities of Foeniculum vulgare Mill., Anethum graveolens L. and Pimpinella anisum L. (Apiaceae) in response to zinc

24 The effect of zinc (Zn) on phenols, antioxidant activities (free radicals’ scavenging activities, 25 inhibition of lipid peroxidation, chelating activity and reducing power), and enzyme inhibition 26 activities of acetylcholinesterase, lipoxygenase and tyrosinase of anise, dill and two cultivars 27 of fennel [Latina” (FL) and “Doux de Florence” (FDF)] aqueous extracts was evaluated for the 28 first time. At the same time, growth and photosynthetic pigment contents were also estimated. 29 A significant decrease in all growth parameters, such as plant height, fresh and dry weights of 30 aerial parts and roots was statistically proved in the presence of 2 mM Zn. An enhancement of 31 Zn accumulation was observed, particularly in aerial part. Exposure to 2 mM Zn induced 32 quantitative changes in the phenols of all tested extracts. In fennel extracts, a decrease of 33 phenol content was observed when treated with Zn, whereas an increase was found in dill and 34 anise extracts submitted to Zn treatment. All samples, either in the presence or absence of Zn, 35 showed antioxidant activity, independent on the tested assay, with the exception of the 36 cultivar FDF (treated and non-treated), which was unable to scavenge NO radicals. Zn-treated 37 dill and anise presented usually higher antioxidant activity. Concerning inhibitory activities 38 against acetylcholinesterase, lipoxygenase and tyrosinase enzymes, both treated and non39 treated FDF did not inhibit acetylcholinesterase activity. Zinc-treated FDF was also unable to 40 inhibit lipoxygenase activity. The remaining species presented capacity for inhibiting those 41 enzymes and tyrosinase as well, but their abilities varied in response to Zn excess. 42 43

Abstract: The effect of zinc (Zn) on phenols, antioxidant activities (free radicals' scavenging activities, inhibition of lipid peroxidation, chelating activity and reducing power), and enzyme inhibition activities of acetylcholinesterase, lipoxygenase and tyrosinase of anise, dill and two cultivars of fennel [Latina" (FL) and "Doux de Florence" (FDF)] aqueous extracts was evaluated for the first time. At the same time, growth and photosynthetic pigment contents were also estimated. A significant decrease in all growth parameters, such as plant height, fresh and dry weights of aerial parts and roots was statistically proved in the presence of 2 mM Zn. An enhancement of Zn accumulation was observed, particularly in aerial part. Exposure to 2 mM Zn induced quantitative changes in the phenols of all tested extracts. In fennel extracts, a decrease of phenol content was observed when treated with Zn, whereas an increase was found in dill and anise extracts submitted to Zn treatment. All samples, either in the presence or absence of Zn, showed antioxidant activity, independent on the tested assay, with the exception of the cultivar FDF (treated and non-treated), which was unable to scavenge NO radicals. Zn-treated dill and anise presented usually higher antioxidant activity. Concerning inhibitory activities against acetylcholinesterase, lipoxygenase and tyrosinase enzymes, both treated and non-treated FDF did not inhibit acetylcholinesterase activity. Zinctreated FDF was also unable to inhibit lipoxygenase activity. The remaining species presented capacity for inhibiting those enzymes and tyrosinase as well, but their abilities varied in response to Zn excess.
Mill., Anethum graveolens L. and Pimpinella anisum L. (Apiaceae) in response to zinc 23 RESPONSE:This proposal made by reviewer 1 was taken into account.  All cases in which % was separated of the number was corrected. 32 pat was corrected and replaced by part.   Reviewer #2: The present manuscript studied the effect of Zn in the antioxidant activity of fennel, dill 55 and anise. Zn is a heavy metal and is also the responsible for the weakness of several soils that cannot 56 be used for agriculture so that is very important to control the amount of this metal. 57 The introduction is very complete as also the other sections, especially the results and discussion that 58 are clear and compared with the available literature, and the authors concluded that the Zn 59 concentration directly affects the antioxidant activity and also the amount of phenolic compounds. 60 This is very interesting given the importance of the safety in the agriculture practices. 61 This manuscript was already revised and after revision the authors already made the requested 62 corrections according to the journals rules. 63 The manuscript can be accepted in the present form. extracts in response to 2 mM Zinc" submitted to Industrial Crops and Products. The study is original 69 and content of the study was studied properly. My comments on the paper were given below.     I suggest to delete Table 8. It is too long and not necessary. The explain of the major points in the   97   table is enough. 98 RESPONSE: Table 8 was deleted from the main document, but at this moment it is a   99 supplementary information. We consider that the information compiled in previous Table 8, 100 now Table S1, is important and must remain.

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We hope that we have addressed correctly all the reviewers and Editor remarks and questions.

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Hoping that everything is in the correct form and looking forward to hearing from you. The effect of zinc (Zn) on phenols, antioxidant activities (free radicals' scavenging activities, 25 inhibition of lipid peroxidation, chelating activity and reducing power), and enzyme inhibition 26 activities of acetylcholinesterase, lipoxygenase and tyrosinase of anise, dill and two cultivars 27 of fennel [Latina" (F L ) and "Doux de Florence" (F DF )] aqueous extracts was evaluated for the 28 first time. At the same time, growth and photosynthetic pigment contents were also estimated. 29 A significant decrease in all growth parameters, such as plant height, fresh and dry weights of 30 aerial parts and roots was statistically proved in the presence of 2 mM Zn. An enhancement of 31 Zn accumulation was observed, particularly in aerial part. Exposure to 2 mM Zn induced 32 quantitative changes in the phenols of all tested extracts. In fennel extracts, a decrease of 33 phenol content was observed when treated with Zn, whereas an increase was found in dill and 34 anise extracts submitted to Zn treatment. All samples, either in the presence or absence of Zn, 35 showed antioxidant activity, independent on the tested assay, with the exception of the 36 cultivar F DF (treated and non-treated), which was unable to scavenge NO radicals. Zn-treated 37 dill and anise presented usually higher antioxidant activity. Concerning inhibitory activities 38 against acetylcholinesterase, lipoxygenase and tyrosinase enzymes, both treated and non-39 4 inflammatory, cytoprotective, hepatoprotective, hypoglycemic, and oestrogenic (Badgujar et 72 al., 2014). 73 Anethum graveolens L. (dill) is an annual and sometimes biennial umbelliferous 74 (Apiaceae) herb, native to south-west Asia or south-east Europe, in the Mediterranean region 75

Determination of growth parameters 115
For each treatment, the fresh and dry weights, and high of roots and aerial part were determined. Fennel, dill and anise treated and non-treated samples (roots and aerial part) were digested 132 by applying the optimized procedure using nitric and perchloric acids. In brief, a 0.05 g of 133 well-powdered samples were added to 5 mL of HNO 3 and HClO 4 (3:1) (v/v) and the mixtures 134 were digested firstly in ambient temperature for 24 h and then with an increased temperature 135 starting from 60°C to 90°C and finally to 105°C until total dissolution. After cooling of the 136 digest, an equal volume of Milli Q water was added to dilute left over acid. Blank solution was 137 prepared by following the same procedure but without samples, only with reagents. The 7 amount of Zinc in the sample solutions was determined by MP-AES (Microwave Plasma 139 Atomic Emission Spectrometry) (Agilent 4200 MP-AES, Santa Clara, CA). 140

Preparation of the extracts 141
The air dried material of fennel, dill and anise species were powdered by using electric 142 mixer grinder and were weighed (5 g for each) and then macerated with distilled water 143 (100 mL) without agitation, at room temperature, for 24 h, filtered, and finally, the filtrate was 144 freeze dried. The absorbance of the reaction mixture was measured at 510 nm. The flavonols and flavones 160 content of the extracts were expressed as quercetin equivalents (mg QE/mL extract)

Flavanone and dihydroflavonol content 162
The total quantification of flavanone and dihydroflavonol compounds was determined 163 according to El-Guendouz et al. (2016). Briefly, 75 µL of sample or standard (naringin) and 164 2 mL DNP (2,4-dinitrophenylhydrazine) (1 g DNP in 2 mL 96% sulphuric acid diluted to 165 100 mL with methanol) were heated at 50 ºC, for 50 min. After cooling at room temperature, 166 175 µL of KOH 10% were finally added to the mixture. The resulting solution was diluted to 167 3 mL with methanol. The absorbance was measured at 486 nm. All determinations were 168 performed in triplicate. The values are expressed as naringin equivalents (mg NE/mL extract). 169

171
This assay was performed using two distinct lipid substrates: egg yolk and liposomes. 172 The TBARS assay is the most commonly used method for measuring lipid peroxidation. 173 The assay was carried out as described by Boulanouar et al. (2013). Briefly, egg yolk 174 (100 mg/mL) (250 µL) homogenates were used as a lipid-rich medium and mixed with acetic 175 acid (750 µL) and the solution of TBA (8 mg/mL) (750 µL) dissolved in sodium dodecyl 176 sulfate (SDS) (11 mg/mL). The method involved heating the reaction mixture for one hour in 177 water bath at 95 °C. After cooling at room temperature, the final volume was adjusted to 2 mL 178 by adding butanol followed by vigorous vortexing and centrifugation for 10 min at 3,000 g. 179 The absorbance of the upper layer was measured at 532 nm against a blank that contained all 180 reagents minus the sample; the percentage of inhibition was calculated as follows: 181 inhibition percentage was plotted against extract concentration (w/v) and IC 50 values were 184 determined (concentration of extract able to prevent 50% of lipid peroxidation). 185 The liposome assay started with the preparation of liposome solution: 0.4 g lecithin in 186 80 mL chloroform (Boulanouar et al., 2013). This solution was dried and submitted to 187 nitrogen flux for 30 s and then to vacuum for at least two hours until complete dryness. In 188 brief, the reaction was initiated by mixing 50 µL of different extract dilution with 100 µL of 189 liposome suspension, Fe Cl 3 (100 µL, 4 mM) and 50 µL of ascorbic acid (0.18 mg/mL diluted 190 to 1/10). After incubation at 37 °C for one hour, 2 mL of TBA solution (0.6%) was added and 191 the resulting mixture was further heated for 10 min at 95 °C. Afterwards, 2 mL butanol was 192 subsequently added and final solution was centrifuged for 5 min at 3,000 g. The absorbance of 193 the supernatant was determined at 532 nm. Tests were carried out in triplicate. The assay was 194 performed as reported above for thiobarbituric acid reactive species (TBARS) method. Our findings show that the accumulation of Zn in the aerial part of fennel (independent on 318 the variety) is mediated by different mechanisms when compared to those of dill, in which the 319 amounts of Zn in the aerial parts are much higher than in roots. According to Chilian et al. 320 (2015) such could be attributed to higher HMA4 activity than that of HMA3, because HMA3 321 protein mediates Zn accumulation in root vacuoles, whereas HMA4 is involved in the metal 322 translocation from root towards the shoot (Lin and Aarts, 2012). Following this reasoning, we 323 could hypothesize that in dill, HMA4 could act more intensely than in fennel or anise, due to the higher amounts of Zn in the aerial part. However, these hypotheses need further 325 confirmation since it was not the aim of the present work. 326  (Table 2), which can suggest that these 340 species are more sensitive to high concentration of Zn than fennel. 341

Effects of Zn on plant growth and its pigment content
Results depicted in Table 2 show that all tested plants remained alive and have grown until 342 the end of treatment, nevertheless a depressing effect caused by Zn exposure in all growth 343 parameters (plant height, root length, fresh and dry weight of aerial parts and roots) was 344 observed. Indeed a strong negative correlation (p < 0.01) between Zn amount and growth 345 parameters was estimated, which confirmed the toxic effect of Zn treatment on samples (Table For all samples, the results presented in Table 1 indicate that the chlorophyll a content 348 (Chla) is lower in treated plants comparing with non-treated samples. The same trend was 349 noted for the chlorophyll b content (Chlb) after the treatment with zinc. The Zn exposure 350 caused a significant decrease of carotenoid contents in all studied samples. The results 351 confirmed that these cultivars of fennel, dill and anise are sensitive to Zn excess. 352 According to the results depicted in Table 4, the measured parameters such as high length, 353 dry weight and fresh weight of roots and aerial part are positively correlated with total 354 chlorophyll and carotenoid contents. 355 Our results also showed a dramatically reduction in carotenoid contents in response to Zn 356 excess as compared to other photosynthetic pigments. In addition, the significant decrease in 357 chlorophyll contents (total chlorophyll (Chla + Chlb), Chla, and Chlb) seems to be more 358  Table 5, the accumulation of phenols and flavonoids depends on the variety. 372

As shown in
Under stressful conditions, the two varieties of fennel plants showed a decrease in phenol and 373 flavonoid contents, whereas in the other species higher accumulation of phenols was detected 374 in the Zn treated plants (dill followed by anise) than in non-treated samples. Data from Table 5  375 show that treatment had a significant increment (p < 0.05) in the amount of dihydroflvonols in 376 the F L as well as in anise, whereas in dill their accumulation in treated plants decreased by 377 two-fold, when compared to the control. 378 The results of the present work show a variability of total phenol contents for the different 379 tested samples. Treated anise and dill samples showed a significant increase of their total 380 phenol concentrations while a reduction in phenolic production was observed for both F DF and 381 F L cultivars exposed to elevated Zn concentration. Thus, it can be argued that the response in 382 terms of phenolic accumulation due to Zn excess is species dependent. Such higher amounts with those obtained in the present work, whereby the effect of metals on phenol accumulation 395 in plants is dependent on the species. Therefore, no significant correlation between aerial part Zn accumulation and phenol content was observed (Table 6)

Zn effects on antioxidant activities of samples 404
Generally, the extracts of anise had higher antioxidant activity than fennel (Table 7). On 405 the other hand, the zinc exposure enhanced the activity in dill and anise extracts, in contrast to 406 the fennel extracts. In this case and in both varieties, zinc had a negative effect on the 407 antioxidant activity (Table 7). Accordingly, our results revealed that treated samples, rich in 408 polyphenols, when compared to the non-treated ones, responded by an enhancement of their 409 capacity for scavenging DPPH, ABTS, nitric oxide and superoxide radicals, as well as 410 through the total antioxidant measured by phosphomolybdenum method. The antioxidant 411 activity correlated well with the phenols' amounts. FRAP assay showed that dill extracts 412 (treated and non-treated), and Zn-treated anise extract possessed the greatest reducing power 413 followed by anise, and fennel (F L and F L Zn). However, treated and non-treated F DF was not 414 able to reduce the iron metal ion. Moreover, the activity was dose-dependent ( Figure 1). 415 In order to discuss our results, we present a reference Table gathering the previous  416 investigations about fennel, anise and dill (Table S1) which is organized as follows: the type 417 of assay studied in the work followed by the plant part used, extraction type as well the 418 solvent used and, finally, the chemical composition for finding a possible relationship 419 between activity and chemical composition. 420 The results found for fennel samples are within the range reported by other authors for 421 extracts obtained by diverse methods, but particularly for DPPH method, and solvents (Table  422   S1). This Table only compiles antioxidant activities, measured through diverse methods, of  423 the aerial parts of fennel, anise and dill as a whole or as parts (leaves, stems, shoots, 424 inflorescences and flowers). The term "Plant material" was also considered. Fruits and seeds 425 were not taken into account, because we considered that we should compare results obtained 426 from the same plant part. In the evaluation of the antioxidant activity, the authors almost 427 always use, at least two assays, although the capacity for scavenging DPPH radicals 428 predominates in practically all works. Beyond the diversity of unities used by the authors for 429 presenting the results, Table S1 permit to show great differences among the results of 430 antioxidant activity, whereby our ones are within the large range found by other authors. Type 431 of extraction and solvent, part of plant used and plant itself can be altogether factors that 432 determine this variability. 433 In the majority of cases, the chemical composition of the extracts was not performed, 434 nevertheless in those in which such was done (nine), practically did not contribute to correlate 435 the activity with the components identified in the extracts. Only in three cases, the authors 436 described a correlation between the antioxidant activity of extracts with some components 437 detected in the same extracts (Faudale et al., 2008;Hossain et al., 2011;Parejo et al., 2004). 438 In these cases, the authors considered that chlorogenic acid isomers, rosmarinic acid, gallic 439 acid and luteolin-7-O-glucoside were determinant in the antioxidant activities found in 440 extracts. 441 Independent on the assay tested, the authors of the works (Table S1) found a correlation 442 between the activities and the concentration of total phenols, such was observed in our 443 investigation (Table 8). In the present work, a correlation was also found between the flavanones and di-hydroflavonols (Table 8). However, the capacity for preventing lipid 446 peroxidation, either using egg yolk or liposomes as lipid substrates, as well as the capacity for 447 scavenging NO free radicals did not correlate with total phenol content. Only the 448 concentrations flavone/flavonols correlate with the capacity for preventing lipid oxidation, 449 when egg yolk was used as lipid substrate, although no correlation had been found when 450 liposomes constitute the lipid substrate of the reaction (Table 8). 451 The antioxidant activity of the aerial parts of anise and dill (Table S1) was also reviewed 452 and much lower references were found when compared to fennel. When the activity unities 453 were presented as IC 50 , it was possible to compare the results of the present work with them. Such results allowed detecting that fennel, independent on the variety, presented a negative 467 effect by decreasing the activity whereas in anise and dill such was not evident.
The accumulation of low molecular phenols by plants in stress or pollution conditions has 469 been reported by diverse authors. According to Mongkhonsin et al. (2016), caffeic acid and 470 rutin were at higher amounts in the leaves of Gynura pseudochina treated with zinc and/or 471 cadmium. However, there was a threshold above which a decrease in the contents of those 472 compounds was observed. In the same work, the authors also reported that higher amounts of 473 rutin and caffeic acid correspond stronger capacity for scavenging DPPH free radicals. to increase their antioxidant activity determined by DPPH radical scavenging and β-carotene 494 bleaching methods, which could be suggested as a protective response of these plants from 495 copper induced damages (Elzaawely et al., 2007). 496 Anise plants under Zn toxicity, with higher amounts of phenolics, revealed a capacity for 497 metal chelating four-fold more efficient than non-treated plants (Table 7). All treated samples, 498 without exception, increase their ability to inhibit lipid peroxidation, particularly when 499 liposomes were used as lipid substrate, in response to Zn stress. These results explain the 500 strong correlation between the effective ability of samples against lipid peroxidation and their 501 Zn accumulation (Table 6). In contrast, the higher accumulation of Zn in all treated samples 502 did not correspond to stronger capacity for scavenging free radicals (DPPH, ABTS, 503 superoxide, NO) in all samples, which may explain the absence of correlation between zinc 504 accumulation and capacity for scavenging some types of free radicals. Zinc is a mineral 505 antioxidant, such as selenium, that does not act on free radicals, but act preventing lipid 506 peroxidation (Prasad et al., 2004). 507

Zn effects on inhibitory activity of enzymes 508
Inhibitory activity of extracts on lipoxygenase (LOX), acetylcholinesterase (AChE) and 509 tyrosinase were performed. 510 Anise extracts exhibited strong LOX inhibitory activity with IC 50 values of 0.015 and 511 0.035 mg/mL for treated and non-treated samples, respectively. F L treated samples (F L Zn) 512 showed the lowest ability to inhibit LOX activity (IC 50 = 0.062 mg/mL), followed by non-513 treated ones (IC 50 = 0.052 mg/mL). The extract from non-treated F DF plants revealed an IC 50 514 value of 0.049 mg/mL, whereas Zn treated plants (F DF Zn) did not present the ability for 515 inhibiting 5-LOX enzyme. The exposure to Zn excess showed insignificant changes in the 516 potency of dill extract as a LOX inhibitor (Table 9). These results implying that Zn application 517 influence the LOX inhibitory capacity of samples but depending on variety and species. 518 The LOX inhibition assay can be considered not only as an indicator of antioxidant but 519 also an indicator of possible anti-inflammatory activity (Albano and Miguel, 2010; 520 Kontogiorgis et al., 2016). These authors studied the antioxidant activity and LOX inhibitor 521 activity of F. vulgare extracts. However, there is no study, at least to our knowledge, focused 522 on fennel and anise Zn treated extracts as enzymatic inhibitors. It has been previously ascribed 523 by Molassiotis et al. (2006) that treatment with boron was associated with an enhancement in 524 LOX activity in leaves and stems of explants of the apple rootstock EM 9 (Malus domestica 525 Borkh). According to these authors, LOX activity may be considered as indicator of oxidative 526 stress. Indeed, it has been suggested that propagation of lipid peroxidation in plants under 527 stress condition induced by higher lipolytic activity on the membrane was associated with 528 stimulation of LOX activity (Lacan and Baccou, 1998). 529 As shown in Table 10, all samples had a significantly moderate ability for inhibiting AchE 530 except F DF samples (treated and non-treated), which were not able to inhibit AchE. Moreover, 531 different trends were observed in the studied varieties after treatment with Zn. Extracts from 532 Zn stressed F L and anise plants revealed higher capacity of AchE inhibition than extracts from 533 the respective non-treated plants, while the opposite occurred with the extracts form dill plants 534 treated and non-treated with Zn. 535 Up to date, searching for new AchE inhibitors derived from natural sources with few side 536 effects is required (Hasbal et al., 2014) but no relevant studies have been reported about 537 anticholinesterase activity of fennel, dill and anise exposed to Zn excess which prompted us to 538 concentrate on the efficiency of our samples for AchE inhibition. Among several stem bark and leaves of Stevia crenata might be attributed to their increased amounts of non-541 enzymatic antioxidants. 542 In this work, although anise and some fennel plants were shown to possess an inhibitory 543 effect against AchE, it was not revealed a significant correlation between AchE inhibitory 544 activity and phenolic compounds including dihydroflavonols, suggesting the contribution of 545 other bioactive constituents against AchE (Table 6). It is noteworthy to refer that there is not a 546 clear trend on the AchE inhibitory activity of tested samples under Zn excess. In fact, no 547 correlation between zinc content and the inhibitory effect of samples against AchE was 548 observed in the present data (Table 6), and to best of our knowledge, scientific information 549 remains absent for better understanding the zinc contributing function in the inhibitory AchE 550 activity. 551 In the present research, all tested extracts exhibited tyrosinase inhibitory activities; however 552 these were higher for dill and anise plants than for both F DF and F L cultivars, which were in the 553 lower range (Table 9). In addition, the inhibitory activity values of dill and anise Zn treated 554 extracts were 1.3 fold higher than those measured for non-treated extracts, suggesting that Zn 555 treatment increased the ability of samples to inhibit tyrosinase enzyme. The extract form dill 556 plants treated with Zn was found as a potent tyrosinase inhibitor. The inhibitory activity of 557 tyrosinase of dill extracts was better than those reported by Orhan et al. (2013). The tyrosinase 558 inhibitory activity, expressed as IC 50 , was negatively correlated with the amounts of total 559 phenols and flavonoids (Table 8), which can be explained by the essential role of hydroxyl 560 group of phenolic compound for forming hydrogen bond with a site of tyrosinase enzyme. 561 There was no correlation between zinc accumulation and antityrosinase activity. 562 Tyrosinase inhibitory activity of kojic acid and its synthetic derivatives is related to the ability 563 to coordinate metals (Lachowicz et al. 2015). Synthetic kojic acid derivatives obtained by 564 these authors had affinity for Fe 3+ , Al 3+ , Zn 2+ , and Cu 2+ . However, the chelating capacity for 565 Zn 2+ , and Cu 2+ was weaker. The absence of correlation between zinc accumulation and 566 antityrosinase activity of extracts may be supported by the finding of Lachowicz et al. (2015). 567 The chelating ability of the compounds present in the extracts with antityrosinase activity for 568 zinc is weak and, consequently, without any supplemental inhibitory effect on tyrosinase 569 activity. 570 571

Conclusions 572
The results of the present work showed that growth parameters and photosynthetic 573 pigment contents were negatively affected by Zn treatment but depending on the plant species 574 and even variety. An accumulation of zinc was observed in both roots and aerial parts of all 575 species studied, though more noticeable in the aerial parts of anise and dill. The plants 576 responded differently to Zn excess exposure, especially in their accumulation of phenols and, 577 therefore, in their abilities for scavenging free radicals and inhibit tyrosinase activity. The 578 accumulation of zinc in plants was only negatively correlated with the IC 50 values of TBARS 579 assay, independent on the lipid substrate used (egg yolk and liposomes), which may indicate 580 the positive role of zinc on the prevention of lipid peroxidation. 581 The extracts of zinc-treated anise and dill plants had higher antioxidant activity and the 582 extracts of anise-treated plants also had higher ability for inhibiting the activity of 583 acetylcholinesterase, lipoxygenase and tyrosinase. This study gives new insights about the 584 effect of Zn excess exposure on the ability of tested extracts against some enzymes but it is 585 important to pay attention about the high accumulation of Zn in the aerial parts of these plants 586 which make their consumption dangerous for human health, due to the toxicity of this metal. 587 On the other hand, the ability of dill and anise plants to accumulate high amounts of Zn raises 588 the hypothesis of their use for bioremediation of soils contaminated with this metal. 589 Table 1 774 Zn content in roots and aerial parts of non-treated and Zn-treated F. vulgare, A. graveolens and P. anisum plants