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Industrial Crops and Products 53 (2014) 65– 70

Contents lists available at ScienceDirect

Industrial Crops and Products

journa l h om epage: www.elsev ier .com/ locate / indcrop

anganese treatment effects on terpene compounds of Cuminumyminum flowers

aryam Ghannadniaa, Raheem Haddada,∗, Fateme Zarinkamarb, Mozafar Sharifib

Department of Agricultural Biotechnology, Faculty of Engineering, Imam Khomeini International University, Qazvin, Islamic Republic of IranDepartment of Plant Biology, Faculty of Biological Science, Tarbiat Modarres University, Tehran, Islamic Republic of Iran

r t i c l e i n f o

rticle history:eceived 23 June 2013eceived in revised form 20 October 2013ccepted 22 October 2013

eywords:uminum cyminum L.umin flowersanganese sprayingonoterpenes

umin organs

a b s t r a c t

The present study was undertaken to identify terpene compounds in different organs i.e. root, shoot,leaf and flower of cumin (Cuminum cyminum L.) as well as the flowers of treated plants with differentconcentrations of manganese (Mn). The plants were sprayed with four different concentrations of Mn(0, 40, 80 and 160 ppm) at (1) both late vegetative and blooming stages, and (2) only at blooming stage.Later, �-pinene, �-pinene, �-phellandrene, m-cymene and �-terpinene compounds were determinedin different organs of untreated plants as well as in flowers of treated plants. Quantity and quality ofthe identified compounds were different in the various plant parts as well as in the flowers of treatedplants. Terpene compounds of the flowers were the same as shoots, but had higher concentration inthe flowers. Notably, �-phellandrene was the only terpenoid compound in the leaves and �-pinene and�-pinene were not detected in the roots. The highest concentration of �-pinene was observed in the

flowers of plants treated with 80 ppm concentration of Mn at blooming stage. According to the results,�-phellandrene and m-cymene increased after treatment with the highest Mn concentration (160 ppm)both at vegetative and blooming stages. Also, �-pinene and �-phellandrene were reduced considerablyin the flowers treated with 40 ppm concentration of Mn at blooming. Overall, the manganese treatmentsat both vegetative and blooming stages increased the monoterpene concentrations of the flowers morethan treatment at the blooming stage alone.

. Introduction

Cumin (Cuminum cyminum L.) is a small, slender, glabrouserbaceous annual plant belonging to the family Apiaceae.he white or pink flowers bloom in small compound umbelsVedamuthu et al., 1994). The seeds come as paired or separatearpel and are 3–6 mm long. In addition of medicinal propertiesAslam et al., 2003; Aruna et al., 2005), antiradical and antimicro-ial properties of cumin seed oil have been reported (Ramadant al., 2012). Cumin also has insecticidal (Ebadollahi et al., 2012),ntifungal (Shahi et al., 2012) and larvicidal (Singha and Chandra,011) activities that can be exploited commercially as it is easilyio-degradable and less toxic to environment. Cumin seeds containolatile oil (2–5%) that imparts the characteristic aroma to the seeds

Behera et al., 2004). The characteristic flavor of cumin is probablyue to dihydrocuminaldehyde and monoterpenes (Weiss, 2002).uminaldehyde, �-pinene, �-pinene, p-cymene and �-terpinene

∗ Corresponding author at: P.O. Box 34149-288, Islamic Republic of Iran.el.: +98 0281 8371165; fax: +98 02813780073.

E-mail addresses: raheemhaddad@yahoo.co.uk, raheemhaddad@ikiu.ac.irR. Haddad).

926-6690/$ – see front matter © 2013 Elsevier B.V. All rights reserved.ttp://dx.doi.org/10.1016/j.indcrop.2013.10.034

© 2013 Elsevier B.V. All rights reserved.

have been usually reported as the main constituents (BettaiebRebey et al., 2012). The terpenes are volatile compounds withstrong flavor that are most often extracted from plant materials.These compounds are distributed in various parts of plants: in flo-rets, fruits, leaves, bark, and roots (Sedlakova et al., 2002). El-Sawiand Mohamed (2002) reported that cumin herb had oil and ter-penoid compounds. The composition of the volatile oil obtainedfrom the cumin herb markedly differed from that of the seed (El-Sawi and Mohamed, 2002). Composition of the essential oil ofcumin depends on many factors such as plant part, harvest-time,extraction method, type of cultivar, geographic origin, and storageconditions (Kan et al., 2007). The production of essential oils notonly depends upon the metabolic state and preset developmen-tal differentiation program of the synthesizing tissue, but also ishighly integrated with the physiology of the whole plant. In general,terpenoids are a predominant constituent of plant essential oils,but many of these oils are also composed of other chemicals likephenypropanoids (Sangwan et al., 2001). Application of micronu-trients, as supplements to macronutrients, has been reported to

have significant effects on herb yields and oil contents of oci-mum, marjoram, mint, and geranium (Sharma et al., 1980; Wahaband Hornok, 1983). Manganese has been shown to be the mosteffective single micronutrient enhancing oil production (Nandi and

6 al Crop

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2

2

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6 M. Ghannadnia et al. / Industri

hatterjee, 1991). This essential trace element plays an importantole in several physiological processes as almost every compart-ent of the cell carries at least one enzyme whose activity is

ependent on Mn. Manganese acts as cofactor for oxidases, dehy-rogenases, and sugar transferases (Crowly et al., 2000; Culottat al., 2005; Keen et al., 2000). Cumin herb and seed showed theifferences in the oil constituents under different treatments ofn. Application of the microelement spray increased the main con-

tituents such as cuminaldehyde, p-cymene, �-terpineol, thymol,nd acoradiene, but other constituents such as �-pinene decreasednder this treatment (El-Sawi and Mohamed, 2002). To the bestf our knowledge, there have been few reports on composition ofhe herb oil of cumin (El-Sawi and Mohamed, 2002). No reportppears to have been published on the terpenoid componentsf cumin separated organs or on the effects of Mn treatmentsith different concentration on terpenoid compounds of cuminowers with a length of 3–4 mm. However, physiological factorsegulating terpene accumulation are poorly known. Given the eco-omic importance of monoterpenes for different industries such as

ragrance, flavor, and pharmaceutical industries, sufficient infor-ation concerning different aspects of essential oil (containing

erpenoid compounds) metabolism in plants such as site of oil pro-uction, ontogeny, photosynthesis, and nutritional relationship isecessary. Also, knowledge of the processes that control monoter-ene biosynthesis and accumulation in plants can be of value in

ncreasing the yields of these commercially valuable natural prod-cts. The present study investigates terpene constituents of cuminoot, shoot, leaf, and flower to illustrate their quantity and qual-ty in each organ and also the changes of these compounds underifferent treatments of Mn in the flowers as the most importanttorage organs of the plant for human consumption.

. Materials and methods

.1. Plant material, growth conditions, and treatments

Seeds of cumin native to Neishabour region (Iran) were sub-ected on water current for 12–15 h. Then, they were surfaceterilized by immersion in 2% (v/v) NaOCl for 3 min and sulfur0% DF fungicide for 2 min, followed by three times rinsing interile water after each step. The seeds were next transferred to

lastic pots containing Klasmann-Deilmann potgrond H peat mossnder equal greenhouse conditions (day/night cycle of 16/8 h, at6 ± 3 ◦C) and irrigated three times per week. At late vegetativetage, the plants were subjected to different concentrations of Mn

Fig. 1. Cumin grown under greenhouse condition at veg

s and Products 53 (2014) 65– 70

(as MnSO4.4H2O; 0, 40, 80 and 160 ppm) as foliar sprays at both latevegetative and blooming stages or at blooming stage only (Table 1).The terpene compounds of different organs were also investigatedin the untreated plants. For terpene extraction analysis of differ-ent organs, samples of root, stem, leaf, and flower (with a length of3–4 mm) were harvested (Fig. 1). To study the Mn content absorp-tion, total aerial parts of the plants from each of the eight treatments(Table 1) were harvested. To investigate the effects of Mn on ter-pene constituents, flowers with a length 3–4 mm were selectedfrom the different treatments 12 h after spraying. After washingwith distilled water, the plant samples were dried at room temper-ature, protected from light, and stored in capped bottles.

2.2. Determination of Mn content in cumin

Aerial parts of the plants in different treatments were rinsedtwice with distilled water. Samples of each treatment were driedfor 24 h at 60 ◦C, weighed, and then ashed in a 480 ◦C oven for16 h. After cooling, the ash was digested with 2 ml HNO3 (65%) andheated to dryness. The sample was then dissolved in 5 ml 3 N HCland brought to volume in a 25 ml volumetric flask using 0.1 N HCl.The Mn contents were determined by atomic absorption system(GVC 902, Australia).

2.3. Terpene extraction from plant materials

Preparation of terpene extracts were performed as describedpreviously (Crocoll et al., 2010) with slight modifications i.e. driedplant materials were ground to a fine powder with mortar andpestle. The powder (50–100 mg) was soaked in 2 ml ethyl acetate:pentane (2:1) for 24 h at room temperature with constant rota-tion. The solution was cleared with activated charcoal for 5 minand dried over a column of 50 mg water-free Na2SO4. The extractswere transferred into a brown capped bottle and stored at 4 ◦C.

2.4. GC–MS analysis of plant volatiles

Constituents of the extracted samples were identified by gaschromatography (Agilent 6890N, USA) coupled to a mass spectrom-eter (Agilent 5973, USA) or a flame ionization detector (FID). Foranalyses 1 �l of pentane or ethyl acetate: pentane (2:1) extracts

were injected with an injector temperature of 230 ◦C. The ter-penes were separated on a HP-5 column: 30 m length, 0.25 mminner diameter, and 0.25 �m film thickness (J and W Scientific,Sanata Clara, CA, USA); GC-program: 40 ◦C for 2 min, first ramp

etative (A) and flowering (B) stages. Bars: 10 mm.

al Crops and Products 53 (2014) 65– 70 67

5hh

2

pwatmlwf

2

iirww

3

3

iMditwbftta(tLaegMtbptp

Fig. 2. Manganese contents of cumin aerial parts in the different treatments. Treat-ments: BA 0, BA 40, BA 80, and BA 160, the plants sprayed with distilled water(0), 40, 80 and 160 ppm of Mn concentrations respectively, at both late vegetativeand blooming stages; A 0, A 40, A 80 and A 160, the plants sprayed with the same,

TA

B

M. Ghannadnia et al. / Industri

◦C min−1 to 175 ◦C, second ramp 90 ◦C min−1 to 250 ◦C, final 3 minold). GC–MS carrier gas: helium at 1 ml min−1; GC-FID carrier gas:ydrogen at 2 ml min−1.

.5. Compound identification

The identity of the extract components was assigned by com-arison of their retention indices relative to (C8–C22) n-alkanesith those of literature or with those of authentic compounds avail-

ble in the laboratory. Further identification was made by matchingheir recorded mass spectra with those stored in the Wiley/NBS

ass spectral library of the GS–MS data system and other pub-ished mass spectra (Adams, 2001). The percentage determination

as based on peak area normalization without using correctionactors.

.6. Statistical analysis

Each sample was prepared by pooling material from at least tenndividual plants to produce individual extract, and each exper-ment was replicated three times. The experimental layout wasandomized block. Statistical analysis was performed using one-ay ANOVA and significant differences among treatment meansere calculated by Duncan’s multiple range test.

. Results and discussion

.1. Effect of Mn treatments on the Mn content of cumin

The results of this study showed that Mn uptake increased withncrease in Mn concentration in treatments up to 80 ppm (Fig. 2).

anganese concentration in the plants that had been sprayed withifferent concentrations of Mn both at late vegetative and bloom-

ng stages showed 1.2–1.3 fold increase compared to the plantsreated only at blooming stage. The highest concentration of Mnas observed in plants treated with 80 ppm concentration of Mn at

oth vegetative and blooming stages while the lowest was recordedor the controls. The rate by which a chemical substance passeshrough the cuticule and, more generally, the epidermal tissues ofhe plants depends on the following factors: (a) the concentrationnd the physical and chemical properties of the sprayed substance,b) the plant species and its nutritional status, and (c) environmen-al conditions (Marschner, 1995; Schonherr, 2001; Schonherr anduber, 2001). Increased accumulation of Mn in the treated plantsre comparable with the results reported for orange (Papadakist al., 2005), Trachyspermum ammi (Abd EL-Wahab, 2008), andrape (Mou et al., 2011). On the other hand, the decrease of plantsn concentration in the treatment with the highest concentra-

ion of Mn (160 ppm) revealed that there was no direct relation

etween concentration of Mn in this treatment compared with thelants tissues (Lidon and Teixeira, 2000). Plants did not exhibit Mnoxicity in any treatment. Toxicity symptoms of Mn in differentlants occur at different levels of tissue Mn contents in the range

able 1bbreviations for different Mn treatments.

Treatment Description

BA 0 Sprayed with distilled water at both late vegetatA 0 Sprayed with distilled water at blooming stageBA 40 Sprayed with 40 ppm concentration of Mn at botA 40 Sprayed with 40 ppm concentration of Mn at bloBA 80 Sprayed with 80 ppm concentration of Mn at botA 80 Sprayed with 80 ppm concentration of Mn at bloBA 160 Sprayed with 160 ppm concentration of Mn at boA 160 Sprayed with 160 ppm concentration of Mn at bl

A, sprayed before and after blooming; A, sprayed after blooming alone.

concentrations of Mn only at blooming stage. Different letters indicate significantdifferences among the, treatments in a multiple range analysis for 95% confidencelevel. Each value is the mean ± S.E. of three, replicates (P ≤ 0.05).

of 400–2000 ppm (Singh and Steenberg, 1997). Since the plantstolerated the minimum concentration of Mn toxicity without anysymptoms (403 �g g−1 DW in BA 80 treatment), it is likely thatcumin is relatively resistant to excess Mn.

3.2. Terpene composition of different organs

Constituents of the extracted terpene were identified andquantified by GC–MS and GC-FID, respectively. The percentagecomposition of the constituents is presented in Table 2. Thechromatographic analyses allowed the identification of differ-ent products representing 85.4–99.46% in the extracts (Table 2).Because of their importance in this research, the monoterpenesidentified such as �-pinene, �-pinene, �-phellandrene, m-cymene,and �-terpinene (constituting 27.5–99.2% of the total compounds)were studied (Table 2). The proportion of the aforementionedfive monoterpenes was different in each plant part, with themajor being �-pinene in flowers (61.39%), �-phellandrene in leaves(27.5%), �-pinene in shoots (21.17%), and �-terpinene in roots(29.66%). The flowers and shoots were rich in all of the terpenoidswith generally higher concentration in the flowers. Notably, �-phellandrene was the only terpenoid compound in the leaves andthere were no �-pinene and �-pinene in the roots (Fig. 3).

The composition of the essential oil varies widely in various

localities (Butkiene et al., 2008). Terpenoid is the most bio-logically “expensive” biosynthesis of all secondary compounds(Langenheim, 1994). Plants cannot maintain high levels of thesedefense substances in all the tissues and organs and at any given

ive and blooming stages

h late vegetative and blooming stagesoming stageh late vegetative and blooming stagesoming stageth late vegetative and blooming stagesooming stage

68 M. Ghannadnia et al. / Industrial Crops and Products 53 (2014) 65– 70

Table 2Composition and content (% in the extract) of �-pinene, �-pinene, �-phellandrene, m-cymene and �-terpinene of different parts of cumin. Ethylacetate: pentane (2:1) extractsprepared from cumin roots, shoots, leaves and flowers. Terpenoid compounds were identified and quantified by GC–MS and GC-FID, respectively.

Compounds Organs

Root Shoot Leaf Flower

Rt (min) % Rt (min) % Rt (min) % Rt (min) %

�-Pinene – – 11.22 1 – – 11.68 2.00�-Pinene – – 12.47 21.17 – – 13.11 61.39�-Phellandrene 13.14 3.73 13.18 2.01 13.13 27.5 14.17 1.93m-Cymene 13.57 19.37 13.94 4.03 – – 14.82 19.64�-Terpinene 13.71 29.66 14.97 2.01 – – 15.92 14.21Total – 52.76 – 30.25 – 27.5 – 99.2Other compounds – 32.64 – 59.85 – 60.2 – 0.26

R

tpAftvepooetci(cpcigttetpvlgrr

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with 40 ppm concentration of Mn at blooming (A 40 treatment)

Final total – 85.4 –

t , Retention time.

ime. Therefore, particular mixtures with high content of toxic ter-enoids are accumulated in the target tissues (Wahid et al., 1997).ccumulation of monoterpenes is confined to the early stages of

ruit development so that young fruits contain high concentra-ions of them (Boumeester et al., 1998). Cumin fruits contain 2–5%olatile oil that imparts the characteristic aroma to the fruits (Kant al., 2007). Most of the oil (containing terpene compounds) isroduced in cumin fruits, although leaves and stems also containil (El-Sawi and Mohamed, 2002). We have reported high levelf terpene (Limonene) synthase gene expression of cumin flow-rs and shoots (Ghannadnia et al., 2011). The close correlation oferpene synthase gene expression and terpene composition indi-ates that transcript regulation of terpene synthase gene is the mostmportant regulatory mechanism controlling terpene compositionCrocoll et al., 2010) in flowers and shoot of cumin. The chemicalomposition of cumin volatile compound varies depending on thelant parts. In addition, �-pinene, �-phellandrene and m-cymeneompounds were identified in the cumin flowers. These results aren agreement with that reported in the previous studies on Anethumraveolens (Radulescu et al., 2010). It seems likely that in flowers,he geranyl pyrophosphate (GPP) pool is large and is not limitingerpene production. This pool of GPP in young tissues may ben-fit the plant by allowing the rapid production of monoterpeneso repel herbivores or attract predators of the herbivores and thusrotect new growth (Lucker et al., 2004). Among all of the majorolatiles detected in cumin, only �-phellandrene was present in theeaves. Our result is consistent with previous studies on Anethum

raveolens L. (Callan et al., 2007; Vokk et al., 2011). Also, our resultsevealed that although �-pinene and �-pinene were absent in theoot components, the amount of �-phellandrene, m-cymene, and

ig. 3. Composition of the terpenoids present in root, shoot, leaf, and flowers3–4 mm in length) of cumin, Monoterpenes were extracted with ethyl acetate:entane (2:1), identified by GC–MS and quantified by GC-FID (see Section 2 foretails).

90.1 – 87.7 – 99.46

�-terpinene were considerable in this organ. This result confirmsthe previous observations by El-Sawi and Mohamed (2002). Thus,flowers are not the only storage organs, but also vegetative parts ofcumin contain terpenoid compounds that may serve as a valuablesource for different usages.

3.3. Effects of manganese treatments on the terpene constituentsof the cumin flowers

Table 3 shows the differences in the terpene constituents ofcumin flowers after the different Mn treatments. All treatmentsshowed the same constituents (Fig. 4). Extract from treated flow-ers with 80 ppm concentration of Mn at blooming stage (A 80treatment) contained the highest proportion of �-pinene, while�-phellandrene and m-cymene concentrations were highest inflowers sprayed with 160 ppm concentration of Mn at both latevegetative and blooming stages (BA 160 treatment). Such incre-ments were statistically significant. Also, treatment with 80 ppmconcentration of Mn at both late vegetative and blooming stages(BA 80 treatment) significantly increased the concentrations of�-pinene and m-cymene compared to the treatment with the sameconcentration at blooming stage alone (A 80 treatment). On theother hand, application of 80 ppm concentration of Mn at bloomingstage (A 80 treatment) significantly increased �-pinene concen-tration compared with all of the other treatments. Treatment

caused not only a reduction in �-pinene and �-terpinene levelsbut also the absence of �-pinene and �-phellandrene compared

Fig. 4. Concentrations for the five main monoterpene components of cumin flowers(3–4 mm) as affected by, different treatments of Mn. Monoterpenes were extractedwith ethyl acetate: pentane (2:1), identified by GC–MS, and quantified by GC-FID.Treatments: BA 0, BA 40, BA 80 and BA 160, the plants sprayed with distilled, water(0), 40, 80 and 160 ppm of Mn concentrations respectively at late vegetative andblooming stages; A 0, A, 40, A 80 and A 160, the plants sprayed with the sameconcentrations of Mn only at blooming stage.

M. Ghannadnia et al. / Industrial Crops and Products 53 (2014) 65– 70 69

Table 3Changes in monoterpene composition (% of the total five monoterpene compounds) under different concentrations of Mn in cumin flowers. Ethylacetate: pentane (2:1)extracts prepared from cumin flowers. Terpenoid compounds were identified and quantified by GC–MS and GC-FID, respectively.

Compounds Treatments

BA 0 A 0 BA 40 A 40 BA 80 A 80 BA 160 A 160

Rt (min) % Rt (min) % Rt (min) % Rt (min) % Rt (min) % Rt (min) % Rt (min) % Rt (min) %

�-Pinene 11.68 1.11e 11.08 1.89b 11.40 1.38d – – 11.36 1.8b 12.54 28.05a 11.35 1.93b 11.69 1.58c�-Pinene 13.11 22.6f 12.50 56.40a 12.82 29.75de 12.82 29.18e 12.79 30.9c 13.14 13.74g 12.79 30.36cd 13.13 32.08b�-Phellandrene 14.17 1.1g 13.33 1.78d 13.90 1.36f – – 13.83 1.6e 13.73 2.08c 13.80 2.52a 14.18 2.23bm-Cymene 14.82 13.9f 13.55 18.05b 14.54 15.13e 14.71 15.86d 14.47 16.5c 13.86 7.89g 14.46 20.97a 14.83 16.60c�-Terpinene 15.92 56.7a 14.21 13.06h 15.63 46.46b 15.76 43.95d 15.59 45.5c 15.91 41.02g 15.59 41.70f 15.95 42.86e

Total 95.41 91.18 94.08 88.99 96.3 92.78 97.48 95.35

T etters8 n co8 stage

wbwtb1rc

citSeopgb

ftpt(t(aaiebrctDatcaDgalogigel

he values (means of three replicates ± S.E.) in the same row followed by different l0 and BA 160, the plants sprayed with distilled water (0), 40, 80 and 160 ppm of M0 and A 160, plants sprayed with the same concentrations of Mn only at blooming

ith spraying with the same concentration at both vegetative andlooming stages (BA 40 treatment). These results are in concertith the previous reports (El-Sawi and Mohamed, 2002). Although

he effects of nutrition on the constituents of essential oil haveeen reported by several investigators (Maximoos, 1985; Ibrahim,989; El-Sawi and Mohamed, 2002), little information is availableegarding the effect of different Mn concentrations on terpenoidompounds of cumin different organs.

Overall, the results of this study showed that spraying with eachoncentration of Mn at both late vegetative and blooming stagesncreased total terpene compounds compared with spraying withhe same concentration at blooming stage alone (Table 3 and Fig. 4).ince �-pinene is a side product of limonene synthase gene (Colbyt al., 1993; Dewick, 1999) the highest concentration of �-pinenebserved in the A 80 treatment in this study is consistent with ourrevious molecular results based on maximum limonene synthaseene expression in treatment with 80 ppm concentration of Mn atlooming stage in cumin (Zarinkamar et al., 2012).

Essential oil biosynthesis is strongly influenced by severalactors. Among the plant nutrients, Mn has important func-ions in plant metabolism, especially in chlorophyll synthesis,hotosynthesis, nitrate reduction, amino acids and protein syn-hesis, activation of different enzymes, phytohormone regulationAmberger, 1974) and also as cofactor for some monoterpene syn-hase (such as limonene synthase) and cyclization of precursorGPP) to the simplest monoterpene (limonene) (Tabata, 2000). In

previous study, we reported that spraying cumin with 40, 80,nd 160 ppm concentration of Mn at both vegetative and bloom-ng stages significantly reduced terpene (limonene) synthase genexpression compared with applying the same concentrations atlooming stage alone (Zarinkamar et al., 2012). In contrast, theesults of current study revealed that spray with the same con-entrations at both stages enhanced the terpene compounds inhe flowers compared with the treatment only at vegetative stage.espite the low level of terpene synthase gene expression, highmounts of terpene compounds could suggest that terpene syn-hesis in the cumin flowers under different treatments of Mnoncentration is regulated at posttranscriptional (translationalnd/or post-translational) levels (Munoz Bertomeu et al., 2008).ecrease in gene transcript levels may result from either loweredene expression or from a decreased stability of the transcripts or

combination of both (Fitzgerald et al., 2008). However, it seemsikely that due to the effects of Mn on metabolism (Amberger, 1974),il and terpene constituents (El-Sawi and Mohamed, 2002), andradual application of Mn at both vegetative and blooming stages

n this study, terpene compounds increased in the plant despiteene expression reduction (Zarinkamar et al., 2012). Production ofssential oils and aromatics from plants is under diverse physio-ogical, biochemical, metabolic and genetic regulations (Sangwan

are significantly different from each other at P ≤ 0.05. Treatments: BA 0, BA 40, BAncentrations respectively at both late vegetative and blooming stages; A 0, A 40, A. Rt , Retention time.

et al., 2001). Results of this study revealed that the chemical compo-sition of cumin terpenoids varies depending on the plant parts. Theyoung fruits as storage organs contain the highest concentrationof the terpenoids. Although the maximum manganese absorption(approximately 404 �g g−1 DW) was observed in the plants treatedwith 80 ppm concentration of Mn at both vegetative and bloomingstages (BA 80 treatment), the flowers of the plants that were treatedwith 160 ppm at both stages (BA 160 treatment) had the great-est concentration of terpene compounds compared with the othertreatments. It appears possible that the rate of Mn absorbed in theBA 160 treatment (approximately 340 �g g−1 DW) was optimumfor terpenoid biosynthesis.

4. Conclusion

Overall, our results revealed that not only flowers but also vege-tative organs of cumin contained terpenoid compounds. Moreover,some of Mn treatments were found to influence the production ofthe terpenoides in the cumin flowers. Considering the economicimportance of these valuable natural products, these results can beuseful for application in different industries. According to the aim,further studies on quantity and quality of ripened seed extracts ofthe treated plants with Mn are needed.

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