The Effects of Zinc Application and Cutting on Growth, Herb, Essential Oil and Flavonoids in Three Medicinal Lamiaceae Plants

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European Journal of Medicinal Plants 12(3): 1-12, 2016, Article no.EJMP.23589 ISSN: 2231-0894, NLM ID: 101583475

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The Effects of Zinc Application and Cutting on Growth, Herb, Essential Oil and Flavonoids in Three Medicinal Lamiaceae Plants

Mona H. Hegazy 1, Fahad M. A. Alzuaibr 2, Abeer A. Mahmoud 3, Hanaa F. Y. Mohamed 3 and Hussein A. H. Said-Al Ahl 1*

1Department of Medicinal and Aromatic Plants Research, National Research Centre, P.O. 12622,33 El Bohouth St., Dokki, Giza, Egypt. 2Department of Biology, Faculty of Science, University of Tabuk, P.O.Box 741, Tabuk 71491, Saudi Arabia. 3Department of Botany (Plant Physiology Section), Faculty of Agriculture, Cairo University, Egypt.

Authors’ contributions

This work was carried out in collaboration between all authors. Authors HAHSAA, AAM, HFYM and FMAA designed the study, performed the statistical analysis, wrote the protocol and wrote the first draft of the manuscript. Authors MHH and HAHSAA managed the analyses of the study and the literature searches. All authors read and approved the final manuscript.

Article Information

DOI: 10.9734/EJMP/2016/23589 Editor(s): (1) Marcello Iriti, Professor of Plant Biology and Pathology, Department of Agricultural and Environmental Sciences, Milan State University, Italy. Reviewers: (1) Saad Farouk Mohamed Hussien Gadalla, Mansoura University, Egypt. (2) Arun Kumar, Hindu Post Graduate College, Zamania, India. Complete Peer review History: http://sciencedomain.org/review-history/13126

Received 9th December 2015 Accepted 2nd January 2016 Original Research Article th Published 30 January 2016

ABSTRACT

Aims: The objective of this study was to investigate the effect of zinc on growth, fresh herb, essential oil and flavonoids content of Dracocephalum moldavica, Hyssopus officinalis and Salvia officinalis plants at two successive cuttings before flowering stage. Study Design: Our study consisted of three experiments. Each study contained 4 treatments, two time of cuttings (90 and 180 days after transplanting) in each plant and two levels of foliar application treatments of Zn EDTA [(Z1) = 0, tap water; (Z2) = 200 ppm)], which was sprayed directly on the plants 4 times at monthly intervals, starting 30 days after transplanting. The design of each experiment was factorial and planned in a complete randomized block

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*Corresponding author: E-mail: [email protected];

Hegazy et al.; EJMP, 12(3): 1-12, 2016; Article no.EJMP.23589

with three replications. Place and Duration of Study: A field experiment was carried out during two successive seasons, 2014 and 2015 at the Agricultural Research and Experimental Station, Faculty of Agriculture, Cairo University, Giza, Egypt. Methodology: Dracocephalum moldavica, Hyssopus officinalis and Salvia officinalis seeds were sown in the nursery in 25 th November, of 2013 and 2014. On February 1, 2014 and 2015 seasons, seedlings were transplanted into the field. Foliar applications of zinc EDTA was sprayed 4 times at monthly intervals. Growth characters and chemical constituent's determinations were carried out at the first and second cuts after 90 and 180 days from transplanting, respectively before flowering. Plant height (cm), number of branches/plant, herb fresh weight (g/plant) was recorded. The content of total flavonoids % was determined in the fresh herb. Essential oil was obtained by hydro distillation and essential oil % was expressed as ml 100 g -1 fresh herb. Chemical composition of the essential oil was studied using GC-MS and compounds were identified based on their mass spectra and literature. Results: Zinc spraying gave the best results of plant height, number of branches, herb fresh weight, essential oil % and total flavonoids % in the vegetative stage than control treatments in the three plants. Also, the second cut gave the best values from these characters in all plants under study compared to the first one. The main compounds of Dracocephalum moldavica essential oil was neral (19.93-20.56%), geranial (22.57-24.56%), geraniol (15.69-17.91%) and geranyl acetate (28.85-29.60%). β-pinene (23.43-24.99%), trans-pinocamphone (16.78-17.97%) and trans- pinocamphone (29.49-29.95%) were found to be the main compounds in Hyssopus officinalis oil. Moreover, α-thujone (13.82-13.92%), 1,8 cineol (17.45-18.83%) and camphor (26.38-27.51%) were considered as major compounds in Salvia officinalis oil. Conclusion: Zinc foliar spray stimulative affect in fresh biomass, essential oil and flavonoids % were observed. It can be concluded that cutting moldavian balm, hyssop and sage plants in the second cut is essential for maximizing the production of fresh herb, essential oil and flavonoids %. These changes could be relevant to the quality of essential oil and its use in certain food, medicinal uses and cosmetic applications.

Keywords: Moldavian balm; hyssop; sage; zinc; foliar spraying; growth; essential oil; flavonoids; GC-MS.

1. INTRODUCTION branching plant investigated primarily for essential oil composition, native to central Asia Medical and aromatic herbs have been essential and is naturalized in eastern and central Europe components of healthcare throughout human [5]. It is frequently consumed as a food additive history [1]. Recently, there has been a general or as an infusion for its organoleptic properties. opinion that synthetic materials that are As an herbal drug, it is used in stomach and liver commonly used in food and drug industry cause disorders, headache and congestion [6]. Herb many diseases, such as cancer. This has led to extract has been used for its antitumer [7], increased global demand for natural and organic antioxidant [8], and antimutagentic properties [9]. forms of medication. The Lamiaceae is one of It is traditionally used as a heart tonic, the most diverse and widespread plant families in reconstituent, sedative, flatulence, vermifuge, terms of ethnomedicine and its medicinal value is diaphoretic, for snakes bites and stings, nausea, based on the volatile oils concentration [2]. The gastroenteritis and as gargle for stomatitis and Lamiaceae plant family is one of the largest fungal infections [10]. In clinical tests, it has been families among the dicotyledons, many species found that a decoction of this herb is effective in belonging to the family being highly aromatic, curing children's pyelonephritis [11]. due to the presence of external glandular structures that produce volatile oil [3]. This oil is Hyssop ( Hyssopus officinalis L.) is an important important in pesticide, pharmaceutical, perennial medicinal plant with small, linear flavouring, perfumery, fragrance and cosmetic leaves and purplish-blue flowers native to central industries [4]. and Southern Europe, Western Asia, and North Africa [12]. It is commonly used in folk medicine. Dragonhead or moldavian balm ( Dracocephalum Hyssop oil may be found as flavour ingredient in moldavica L.) is an annual herb, erect, bushy, many food products, cosmetics and perfumes

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[13]. Hyssop oil has been used as a healing herb their constituents as well as total flavonoids of to alleviate digestive disorders, cure laryngitis, or dragonhead, hyssop and sage plants and time of accelerate wound healing in Turkish folk cuttings for better plant growth and chemical medicine. It relaxes peripheral blood vessels and composition for each plant. promotes sweating. It is also used as an expectorant, carminative, anti-inflammatory, 2. MATERIALS AND METHODS anticatarrhal, and antispasmodic [14]. As a medicinal herb, hyssop is used in viral infections 2.1 Plant Material and Optimization of such as colds, coughs, sore throats, bronchitis Growing Conditions and asthma, nervous disorders and toothache [13]. The oil is antimicrobial, antifungal, Our study consisted of three experiments were antiseptic, antioxidant mildly spasmolytic, conducted in 2014 and 2015 at the Agricultural antiviral, antiprotozoal and anticancer [15-18]. Research and Experimental Station, Faculty of

Agriculture, Cairo University, Giza, to estimate Sage ( Salvia officinalis L.) is a perennial, the effect of foliar applications of zinc on evergreen subshrub, with woody stems, grayish dragonhead, hyssop and sage plants. The leaves, and blue to purplish flowers. It is native to design of each experiment was factorial and the Mediterranean region, being currently planned in a complete randomized block with cultivated in various countries [19,20]. It is one of three replications. Each study contained 4 the oldest medicinal plants and also has been treatments, two time of cuttings (90 and 180 days used for a long time in folk medicine as after transplanting) in each plant and two levels medication against fever, rheumatism, of foliar application treatments of Zn EDTA [(Z1) perspiration, sexual debility, and in the treatment = 0, tap water; (Z2) = 200 ppm)], which was of chronic bronchitis, as well as mental and sprayed directly on the plants 4 times at monthly nervous diseases [21]. Sage is largely used as a intervals, starting 30 days after transplanting. savory food flavoring either as dried leaves or essential oil [22]. Sage leaves and its essential Seeds of the three plants were sown in the oil possess carminative, antispasmodic, th antiseptic, astringent, and antihidrotic properties nursery on 25 November, of 2013 and 2014. [20]. Essential oil is externally used for Uniform seedlings were transplanted into the inflammations and infections of the mucous field, on February 1, during 2014 and 2015 membranes of throat and mouth (stomatitis, seasons, in 3×3.5 m plots, rows, with 60 cm a gingivitis, and pharyngitis). Internally, the part and and 20 cm between the seedlings. The essential oil is used for dyspeptic symptoms and seedlings were thinned 45 days after excessive perspiration [20]. transplanting to leave one plants per hill. The mean values of physical and chemical analyses Zinc (Zn) is an essential micronutrient for the of the soil were determined according to Jackson normal healthy growth and reproduction of [28]. The soil texture was Clay loam, having the plants, animals and humans [23]. In plants, Zn following physical composition: 39% sand, 25.0% plays a key role as a structural constituent or silt, 36.0% clay and 0.8% organic matter. The regulatory co-factor of a wide range of different results of soil chemical analysis were: pH = 7.9; enzymes and proteins in many important E.C (mmohs/cm) = 1.3 and zinc = 1.0 ppm. biochemical pathways. These roles include carbohydrate metabolism (both in photosynthesis 2.2 Growth Characters and Chemical and in the conversion of sugars to starch), Constituents protein metabolism, auxin (growth regulator) metabolism, pollen formation, the maintenance of Growth characters and chemical constituent's the integrity of biological membranes, the determinations were carried out at the first and resistance to infection by certain pathogens [24- second cuts after 90 and 180 days from 26]. Therefore, many important physiological transplanting, respectively before flowering. The functions of Zn are unable to operate normally in following data were recorded, Plant height (cm), Zn-deficient plants, and plant growth would be number of branches/plant, herb fresh weight adversely affected [27]. (g/plant). The content of total flavonoids in the fresh herb was determined according to the The main objective of the present investigation method given by Boham and Kocipai [29].The was to study the effect of foliar application of zinc essential oil percentage was determined in the on the growth, herb yield and essential oil and fresh herb using a modified Clevenger apparatus

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according to Guenther [30]. Essential oil sprayed by zinc in the first season. Also, there percentage was determined and expressed as was a significant increased in the total flavonoids (%). % between plants sprayed by zinc and nonsprayed plants in the two seasons of hyssop 2.3 GC/MS Analyses Conditions plant. Moreover, there was a significant increment in the plant height between zinc The GC-Ms analysis of the essential oil samples treatment and nonsprayed plants in the second was carried out in the first season using gas season in sage plant. Also, zinc led to significant chromatography-mass spectrometry instrument increased in total flavonoids % compared to stands with the following specifications. control sage in the two seasons. The interaction Instrument: a TRACE GC Ultra Gas treatments between zinc and time of cuts Chromatographs (THERMO Scientific Corp., resulted in a significant increment of plant height USA), coupled with a THERMO mass in the second season and total flavonoids in sage spectrometer detector (ISQ Single Quadrupole plants for both seasons. But, the interaction Mass Spectrometer).The GC/MS system was treatments were not significant in the equipped with a TG-WAX MS column (30 m x dragonhead and hyssop plants. 0.25 mm i.d., 0.25 µm film thickness). Analyses were carried out using helium as carrier gas at a The stimulatory effect of zinc was recorded [31- flow rate of 1.0 mL/min and a split ratio of 1:10 34]. Zinc is a component of carbonic anhydrase, using the following temperature program: 40°C as well as several dehydrogenases and auxin for 1 min; rising at 4.0°C/min to 160°C and held production which in turn enhanced the elongation for 6 min; rising at 6°C/min to 210°C and held for processes, besides the function of zinc in CO 2 1 min. The injector and detector were held at assimilation. Consequently, the fresh and dry 210°C. Diluted samples (1:10 hexane, v/v) of 0.2 weights of herb could be increased [24]. µL of the mixtures were always injected. Mass spectra were obtained by electron ionization (EI) Results from Tables 1-3 also indicated that at 70 eV, using a spectral range of m/z 40-450. harvested all plants in the second cut were much Most of the compounds were identified using two superior in plant height, number of branches, different analytical methods: relative retention herb fresh weight, oil and total flavonoids time and mass spectra (authentic chemicals, percentages compared with first cut. However, Wiley spectral library collection and NSIT library). the differences were significant in plant height, branches number, herb fresh weight, oil % and 2.4 Statistical Analysis total flavonoids % when compared first cut with second cut in dragonhead, hyssop and sage plants in both seasons. It can be noticed from Growth and the essential oil content in the two Tables 1-3 that, Dracocephalum moldavica gave cuts were analyzed with the analysis of variance the highest value of plant height followed by (ANOVA) using JMP 10 program (SAS Institute, Hyssopus officinalis and then Salvia officinalis , NC, USA). The mean values of treatments were also Dracocephalum moldavica gave the highest compared using Tukey’s HSD test. Values value of herb fresh weight followed by Salvia accompanied by different letters are significantly officinalis and then Hyssopus officinalis in the different at p ≤0.05. two cuts during both seasons. However,

Hyssopus officinalis gave the highest values of 3. RESULTS AND DISCUSSION branches number and total flavonoid % followed by Salvia officinalis and then Dracocephalum 3.1 Growth Characters and Chemical moldavica , but Salvia officinalis gave the highest Constituents value of essential oil % followed by Hyssopus officinalis and then Dracocephalum moldavica in Data in Tables 1, 2 and 3 revealed that Zn the two cuts during both seasons. It has been treatment had a considerable effect on plant reported that yield and its components in plants height, branches number, herb fresh weight, in general is primarily related to their genetic, essential oil % and total flavonoids of climate, edaphic, and their interaction etc., [35- dragonhead, hyssop and sage plants in the two 38]. Also, Said-Al Ahl and Mahmoud [33] cuts during both seasons compared with control reported that harvested Ocimum basilicum plants of each plant. For dragonhead, there was a in the second harvest gave the highest plant significant increment in the plant height and herb height, number of branches, fresh matter yield as fresh weight between with or without plants well as essential oil than those of first cut. Also,

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Said-Al Ahl et al. [39,40] in two experiments on Song et al. [48] found that, Zn treatments oregano plants emphasized that, harvested in enhanced the accumulation of total flavonoids of the second cut gave the best result of herb fresh Vitis vinifera cv. Merlot. The same result had weight and essential oil content compared to the obtained by [49]. In Zn-deficient plants, the first cut. The results of two cuttings indicated that activity of carbonic anhydrase (CA), ribulose 1,5- dragonhead performed better in second than in biphosphate carboxylase (RuBPC) and the first cutting [41]. This may be resulted from content of chlorophyll decrease, leading to a suitable climatic conditions and state of maturity decline of photosynthesis [25,50,51]. Meanwhile, during the second cutting period. The yield and the translocation of sucrose from leaves to fruit is quality of essential oil produced depends on also impaired [25]. The promotion effect of various intrinsic and extrinsic factors. Intrinsic foliage sprayed zinc sulfate on photosynthesis of factors comprise all internal hereditary Zn-deficient vines at the late developmental characters. Extrinsic factors affecting the stages was possibly the origin of the initiation of production of essential oil in plants are soil, the change of other parameters considering the nutrition and water supply, climate relationship between photosynthesis, sugar (light/temperature), maturity, post harvest metabolism and phenolic metabolism. treatment and distillation [42]. Penka [43] showed that the formation and accumulation of essential 3.2 Essential Oil Composition oil in plants was explained as due to the action of environmental factors. It might be claimed Table 4 shows the data belonging to qualitative that the formation and accumulation of and quantitative constituents of essential oils essential oil was directly dependent on perfect distilled from the dragonhead, hyssop and sage growth and development of the plants producing herbs before flowering stage treated with zinc oils. applications and collected from the second cut during the season of 2014. Comparison of the The increase in essential oil due to zinc was also analytical data of the oils revealed marked found by [31-34,44]. From previous studies, differences in qualitative and quantitative biosynthesis of secondary metabolites is not only composition in the different three plants. controlled genetically but it also is affected Considering the main components of strongly by environmental influences [45]. In line dragonhead oil were characterized by high with the foregoing, environmental variables affect contents of neral (19.93-20.56%); geranial essential oil, Marschner [24] found that zinc is an (22.57-24.56%); geraniol (15.69-17.91%) and essential micronutrient that acts either as a metal geranyl acetate (28.85-29.60%). Whereas, β- component of various enzymes or as a pinene (23.43-24.99%), trans-pinocamphone functional, structural, or regulatory cofactor (16.78-17.97%) and trans-pinocamphone (29.49- associated with saccharide metabolism, 29.95%) in hyssop oil. Moreover, α-thujone photosynthesis, and protein synthesis. Carbon (13.82-13.92%), 1,8cineol (17.45-18.83%) and dioxide and glucose are precursors of camphor (26.38-27.51%) in sage oil are monoterpene biosynthesis, saccarides are also a considered as major compounds (more than source of energy and reducing power for 10%). terpenoid synthesis. As zinc is involved in photosynthesis and saccaride metabolism and as Said-Al Ahl et al. [52] mentioned that geranyl CO 2 and glucose is the most likely sources of acetate (28.81, 27.02 and 27.81%); geranial, carbon utilized in terpene biosynthesis, the role citral a (20.42, 19.37 and 19.74%); neral, citral b of zinc in influencing essential oil accumulation (18.36, 17.85 and 17.86%) and geraniol (12.66, seems particularly important [46]. 16.05 and 16.68%) which considered as the main compounds of volatile oil extracted from To our knowledge, both phenolics and Zn has Dracocephalum moldavica fresh herb. However, been the subject of a number of studies. geranyl acetate compound contents were the However, very little work has been done to highest in the volatile oil, followed by geranial, investigate the relationship between biosynthesis then neral and finally geraniol in all harvest dates of phenolics and application of Zn fertilizer. Due in Egypt. Also, Janmohammadi et al. [53] found to the fact sucrose is a positive regulator of the that geranial, geraniol and geranyl acetate was biosynthesis of phenolics, particularly flavonoids the major components in dragonhead grown in [47], the improvement of Zn treatment on two locations in Iran. Omidbaigi et al. [54] noticed photosynthesis and sugar accumulation could that geranyl acetate, geranial, neral and geraniol possibly enhance the biosynthesis of flavonoids. were major components in Iran.

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Table 1. Effect of foliar spray with zinc, number of cutting on Dracocephalum moldavica plant

Studied characters Without zinc application With zinc application 1st season 2nd season 1st season 2nd season Ist cut 2nd cut Ist cut 2nd cut Ist cut 2nd cut Ist cut 2nd cut Plant height (cm) 75.7±2.3B 87.7±1.8A 78.0±2.5B 89.3±0.9A 85.5±2.0A 94.5±2.5A 79.9±1.6B 91.3±2.3A Branches number/plant 5.3±0.9A 6.7±0.3A 5.3±0.3B 6.3±0.3AB 5.7±0.3A 6.7±0.3A 6.3±0.3AB 7.3±0.7A Herb fresh weight (g/plant) 141.3±10.2B 152.0±11.7AB 165.0±10.4A 192.0±7.6A 149.8±5.2AB 189.0±5.9A 168.3±9.2A 185.7±3.03A Essential oil % 0.05±0.003B 0.11±0.003A 0.05±0.0D 0.10±0.003B 0.06±0.0B 0.11±0.0A 0.06±0.003C 0.12±0.003A Total flavonoids % 0.45±0.01C 0.53±0.03AB 0.44±0.003C 0.55±0.02AB 0.47±0.01BC 0.59±0.003A 0.50±0.03BC 0.59±0.01A

Table 2. Effect of foliar spray with zinc, number of cutting on Hyssopus officinalis plant

Studied characters Without zinc application With zinc application 1st season 2nd season 1st season 2nd season Ist cut 2nd cut Ist cut 2nd cut Ist cut 2nd cut Ist cut 2nd cut Plant height (cm) 51.7±1.2C 57.7±1.8AB 49.0±0.6B 58.3±1.2A 53.8±0.9BC 60.7±0.3A 50.5±0.3B 63.0±2.5A Branches number/plant 8.3±0.7A 10.7±0.9A 8.7±0.3B 11.0±0.6A 9.3±0.3A 11.3±0.7A 9.0±0.0B 12.0±0.6A Herb fresh weight (g/plant) 17.0±0.6B 24.5±2.7AB 19.6±0.4B 28.7±0.9A 21.0±2.1AB 27.0±2.1A 21.7±1.2AB 29.7±3.5A Essential oil % 0.16±0.01B 0.24±0.01A 0.18±0.01B 0.25±0.02A 0.18±0.003B 0.25±0.01A 0.19±0.01B 0.27±0.02A Total flavonoids % 1.07±0.03B 1.14±0.03AB 1.03±0.03B 1.12±0.0AB 1.17±0.03AB 1.25±0.03A 1.13±0.03AB 1.22±0.03A

Table 3. Effect of foliar spray with zinc, number of cutting on Salvia officinalis plant

Studied characters Without zinc application With zinc application 1st season 2nd season 1st season 2nd season Ist cut 2nd cut Ist cut 2nd cut Ist cut 2nd cut Ist cut 2nd cut Plant height (cm) 30.3±1.8B 37.7±1.5AB 28.3±1.7B 38.3±0.3A 31.9±0.9AB 38.5±2.3A 36.3±1.9A 40.0±0.6A Branches number/plant 8.0±0.6A 9.7±0.3A 7.7±0.3B 9.0±0.0AB 8.7±0.3A 10.3±0.9A 8.0±0.0AB 9.7±0.7A Herb fresh weight (g/plant) 30.2±1.6C 43.0±3.5AB 27.7±1.8B 38.0±1.5AB 33.7±0.9BC 45.7±2.6A 29.0±0.6B 41.0±4.4A Essential oil % 0.33±0.01B 0.47±0.01A 0.32±0.02B 0.49±0.01A 0.34±0.01B 0.50±0.01A 0.32±0.0B 0.49±0.01A Total flavonoids % 0.89±0.01C 0.98±0.02B 0.88±0.02C 1.05±0.03B 0.92±0.02BC 1.18±0.02A 0.90±0.0C 1.17±0.02A

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Table 4. Principal constituents of three Lamiaceae essential oils affected by zinc foliar spray

Compound 2nd season Without zinc application With zinc application Dracocephalum moldavica Hyssopus officinalis Salvia officinalis Dracocephalum moldavica Hyssopus officinalis Salvia officinalis α-thujone - - 13.82 - - 13.92 β- thujone - - 5.96 - - 6.24 β-pinene 0.11 23.43 0.27 0.29 24.99 0.27 β -myrcene 0.17 - - 0.14 - - limonene 0.19 3.03 3.54 0.10 4.40 3.98 1,8-cineole - - 18.83 - - 17.45 linalool 1.97 0.95 0.20 2.03 1.03 0.28 α-phyllandrene 0.09 - - 0.12 - - β-phellandrene - 2.41 - - 3.15 - sabinene 0.10 - - 0.15 - - γ-terpinene 0.08 - - 0.12 - - caryophyllene 0.58 1.68 1.34 0.92 2.03 1.06 β -ocimene 0.29 1.66 - 0.44 0.95 - γ-elemene - 0.20 - 0.22 camphor - - 27.51 - - 26.38 trans-pinocamphone - 16.78 - - 17.97 - cis-pinocamphone - 29.95 - 29.49 - α- humulene - 0.48 4.59 - 0.49 4.89 α-terpineol - 0.45 1.67 - 0.38 1.50 borneol - - 3.43 - - 3.06 neral 20.56 - - 19.93 - - geranial 22.57 0.15 - 24.56 0.13 - nerol 1.49 0.20 - 2.31 0.26 - geraniol 15.69 0.32 - 17.91 0.28 - bicyclogermacrene 0.49 1.45 - 1.11 3.79 - α-terpinyl acetate - - 2.02 - - 2.54 geranyl acetate 28.85 - 0.21 29.60 - 0.32 myrtenol - 1.33 0.34 - 2.70 0.26 carvacrol 0.21 - 0.95 0.19 - 0.67 γ-selinene - - 3.94 - - 3.73 α-elemol - 7.96 - - 6.85 - caryophyllene oxide 0.22 0.23 3.92 0.24 0.38 3.86 spathulenol 0.06 0.29 0.47 0.11 0.26 0.58 humulene epoxide - - 1.12 - - 1.02 manoyl oxide - - 5.59 - - 5.46 β-selinene - - 0.45 - - 0.67

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Veres et al. [55] reported that hyssop essential Other compounds were considered as traces, oil could be categorised depending upon their such as β-pinene; β -myrcene; limonene; percentage composition of β-pinene, limonene, α-phyllandrene; sabinene; γ-terpinene; pinocamphone, and iso-pinocamphone. caryophyllene; β-ocimene; bicyclogermacrene; Figueredo et al. [56] revealed that the major carvacrol; caryophyllene oxide and spathulenol in constituents of Hyssopus officinalis grown in agonhead oil. α- humulene; α-terpineol; geranial; Turkey were pinocarvone (29.2%), trans- nerol; geraniol; caryophyllene oxide and pinocamphone (27.2%), β-pinene (17.6%), cis- spathulenol in hyssop oil. β-pinene; linalool; γ- pinocamphone (4.7%) and myrcene (2.92 %). elemene; geranyl acetate; myrtenol ; carvacrol; Said-Al Ahl et al. [57] revealed that the major spathulenol and β-selinene in sage oil were constituents of Hyssopus officinalis oil were cis- considered as traces (less than 1%), as it was pinocamphone (26.85%), β-pinene (20.43%), cleared from the Table 4. trans-pinocamphone (15.97%) α- elemol (7.96%), durenol (3.11%), β-phellandrene Essential oil productivity is eco-physiologically (2.41%), caryophyllene (2.34%), (E)-2,6- and environmentally friendly. These and other dimethyl-1,3,5,7-octatetraene (2.27%), 3(10)- aspects of the modulation of essential oil caren-4-ol,acetoacetic acid ester (2.14%), production are presented, along with a brief bicyclogermacrene (1.83%), myrtenol (1.73%), outline of the current concept of the relevant germacrene-D (1.68%), limonene (1.56%), γ- biosynthetic mechanisms [61]. Results of eudesmol (1.36%) and linalool (1.08%). previous studies indicated that altitude is the most important environmental factor influencing Good quality sage oils contain a high percentage on oil content in Origanum vulgare ssp. hirtum (> 50%) of epimeric α- and β- thujones and a low and Thymbra spicata var. spicata L. [62] and, the proportion (< 20%) of camphor [20]. Raina et al. highest values of essential oils were recorded at [58] showed that sage had oil rich in thujone low altitudes. Unsuitable environmental content which ranged from 41.31 to 47.51%. α- conditions may limit photosynthesis in plants and thujone considered as major constituent, varying alter nutrient uptake and carbon, sugar, amino from 36.06 to 40.10% in sage oil. The acid and inorganic ion fluxes. constituents α-pinene, β-pinene, 1,8 cineole, α- thujone, β-thujone, camphor, borneol, α- In general, the climate favorable and optimal humulene, viridiflorol, and manool account at environmental conditions for essential oil least 81% of weight reported [59]. Said-Al Ahl et accumulation are important. Different agronomic al. [60] reported that camphore (23.38%), α- practices were observed and optimal conditions thujone (22.82%), sclareol (10.46%), β-thujone for oil production reflect the natural climate (9.96%), 1,8-cineole (7.83%), γ-selinene conditions and the best agronomic practices in (7.73%), α-humulene (5.59%), caryophyllene, medicinal and aromatic plants [63]. (3.16%), borneol (3.06%), limonene (1.74%) and Environmental factors, including cultivation, humulene epoxide (1.02%) which represents the temperature, moisture, and soil, appear to have main compounds of volatile oil extracted from an impact on volatile compound production in Salvia officinalis fresh herb. medicinal and herb crops [64-66]. The concentration of individual volatile compounds in Minor compounds like; linalool (1.97-2.03%) and Thymus piperella responds to a variety of nerol (1.49-2.31%) compounds in dragonhead environmental factors independently of other oil; limonene (3.03-4.40%), linalool (0.95- compounds in the whole oil [64]. In fennel, there 1.03%), β-phellandrene (2.41-3.15%), were cultivar-specific responses to planting dates caryophyllene (1.68-2.03%), β-ocimene (0.95- and, as expected, cultivar-specific chemical 1.66%), bicyclogermacrene (1.11-1.45%), profiles in the essential oils [67]. myrtenol (1.33-2.70) and α-elemol (6.85-7.96%) in hyssop oil as well as, β-thujone (5.96-6.24%), The results in Table 4 show that zinc treatment limonene (3.54-3.98%), caryophyllene (1.06- gave the highest content of ( α-thujone, β-pinene, 1.34%), α-humulene (4.59-4.89%), α-terpineol trans-pinocamphone, geranial, geraniol and (1.50-1.67%), borneol (3.06-3.43%), α-terpinyl geranyl acetate); However, control treatment acetate (2.02-2.54%), γ-selinene (3.73-3.94%), gave the same result of (1,8-cineole, cis- caryophyllene oxide (3.86-3.92%), humulene pinocamphone and neral) in the three plants epoxide (1.02-1.12%) and manoyl oxide (5.46- under study. Said-Al Ahl and Omer [33] indicated 5.59%) in sage oil are considered as minor that linalool was increased in the herb and seeds compounds (less than 10% and more than 1%). of coriander. But, linalool and 1,8-cineol was

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