Plant Growth and Essential Oil Content and Composition of Satureja Hortensis L. Cv. Saturn in Response to Calcium Carbonate and Nitrogen Application Rates

Full Length Research Paper

Plant growth and essential oil content and composition of Satureja hortensis L. cv. Saturn in response to calcium carbonate and nitrogen application rates

Hasan Mumivand 1, Mesbah Babalar 1, Javad Hadian 2* and Mohammad Fakhr-Tabatabaei 1

1Department of Horticultural Sciences, Faculty of Agriculture, University of Tehran, Karaj 31587, Iran. 2Medicinal Plants and Drug Research Institute, Shahid Beheshti University, G. C., Evin, Tehran, Iran.

Accepted 2 September, 2010

Satureja hortensis L. (Summer savory) is one of the most important medicinal and spice plants cultivated in many parts of the world. A field experiment was conducted to study the effects of nitrogen (0, 50, 100 and 150 Kg.ha -1) and calcium carbonate (0, 5 and 10 t.ha -1) application rates on the growth and essential oil content and composition of S. hortensis cv. Saturn. Nitrogen (N) fertilization increased plant height, fresh and dry weights of herb, stem diameter, mean of Leaf Area (LA), drug yield and leaf N content as well as essential oil yield. Plant fresh and dry weights and mean of LA were increased by -1 2+ CaCO 3 up to 5 t. ha . In addition, CaCO 3 application positively affected the leaf Ca content and essential oil content and yield, but plant width and leaf N content were negatively affected. The interaction effect of N and CaCO 3 application was significant for leaf N and essential oil contents. Gas chromatography-flame ionization detector (GC-FID) and gas chromatography mass spectrometry (GC- MS) analyses of essential oil showed that the chemical compositions of S. hortensis essential oil did not change due to N fertilization while CaCO 3 application increased the relative percentages of carvacrol, γ-terpinene and β-bisabollene.

Key words: Satureja hortensis L., essential oil, calcium carbonate, nitrogen, carvacrol.

INTRODUCTION

The genus Satureja L. (Labiatae, Mentheae) comprises world food, drink and perfume industries (Sefidkon et al., over 30 species with wide distribution in the 2006; Skocibusic et al., 2006). The essential oil of S. Mediterranean region (Hadian et al., 2008). Among them, hortensis possesses many activities such as antioxidant, many are used as valuable medicinal and spice plants antibacterial and antifungal (Gulluce et al., 2003; Sahin et worldwide. S. hortensis L. (summer savory) is an annual al., 2003). Main essential oil constituents are phenolic aromatic plant with linear to linear-oblanceolate leaves compounds, carvacrol and thymol, as well as γ-terpinene, and white to pale red flowers, which are born in erect p-cymene, β-caryophyllene, linalool and other terpenoids stems (Rechinger, 1982). In folk medicine, Satureja (Sahin et al., 2003; Zawislak, 2008). hortensis is used as stomachic, stimulant, carminative, In comparison with other crops, the concentration of expectorant, aphrodisiac, antispasmodic and main minerals such as Ca 2+ , Mg 2+ and Zn 2+ in the leaves antidiarrhoeal (Zargari, 1970; Hajhashemi et al., 2000). In of S. hortensis is very high making it as a potential source addition, summer savory has wide application in the of dietary minerals (Ozcan, 2004). Beside, adaptability to harsh environmental conditions, high yield and short growing period make S. hortensis as a valuable alternative crop in agriculture (Hadian et al., 2008). *Corresponding author. E-mail: [email protected]. Tel: In the recent years, the interest of growing herbs such +98-21-29903025. Fax: +98-21-29903023. as savory as alternative crops is highly increased 1860 J. Med. Plant. Res.

Table 1. Some physical and chemical properties length and width of each plot were 2.1 and 1.8 m, respectively (total of the soil of experimental field. area = 3.78 m 2). A factorial experiment based on the Randomized Complete Block Design with three replications was used. N fertilizer was applied at different levels of 0 (unfertilized), 50, 100 and 150 Soil properties Content -1 Kg. ha , as NH 4(NO 3). These fertilizers were divided into two equal Total N (%) 0.09 splits and added at first and fifth weeks from transplanting. Thirty Available K (ppm) 696 months prior to transplanting, CaCO 3 treatment was added as -1 Available P (ppm) 104 slaked lime at different levels of 0 (control), 5 and 10 t. ha by mixing properly with soil (up to 30 cm depth). During the growing Ca (meq/lit) 20 period, the plants were weekly irrigated and kept free of weeds by pH 6.4 hand hoeing. The chemical properties of the soils treated with EC (ds/m) 1.26 different levels of CaCO 3 were analyzed after harvest (Table 2). Organic C (%) 0.93 Organic matter (g/Kg) 6.25 Sampling and traits analysis Textural class Clay loamy

Lime (%) 8.7 Eight plants were sampled from the central part of each plot on the October 12th (71 days after transplanting) at full bloom. Following parameters were recorded for each sample: plant height and width, 2 stem diameter, fresh and dry weights of herb and drug yield per m (Prohens et al., 2003). Production of essential oil and its (leaf and flower fractions which are oil bearing parts of the plant). The mean leaf area (LA) per plant was measured (by 10 leaves) composition in plants is mainly dictated by the combined using a leaf area meter ( ∆T England). In addition, total N and Ca 2+ influences of both genetic factors and cultivation contents of leaves were determined for each treatment according to conditions such as climate, plant density, the use of kjeldahl (Helrich, 1990b) and atomic absorption spectrophotometric fertilizers, etc (Supanjani et al., 2005b; Salmasi et al., (Helrich, 1990a) methods, respectively. 2008). Nitrogen and calcium are known as two of the most important nutrients for plant production and have critical roles in cell structures and plant metabolism Essential oil isolation (Supanjani et al., 2005b; Ashraf et al., 2006; Dordas and Sioulas, 2008). The effects of nitrogen fertilizer on the The aerial parts of harvested plants were air-dried in the shade, and then flower heads and leaves were subjected to hydrodistillation for growth and essential oil content and composition of 3 h using a Clevenger-type apparatus according to the European different plants species have been previously reported Pharmacopeia method (Pharmacopee Europeene, 1997). The (Baranauskiene et al., 2003; Ashraf et al., 2005; Yang et obtained oils were dried over anhydrous sodium sulfate and stored al., 2005; Sifola and Barbieri, 2006; Martins et al., 2007). at 4°C prior to analysis. The essential oil content (v/w) and yield for Also, the positive effects of calcium application on the each plot were determined. essential oil content and composition of several medicinal plants are well documented (Suh and Park, 2000; Lee Gas chromatography-flame ionization detector and gas and Yang, 2005; Supanjani et al., 2005a, b). With regard chromatography-mass spectrometry analysis to the importance of N and Ca on the growth and secondary metabolism, we studied the impacts of Gas chromatography-flame ionization detector (GC-FID) analysis nitrogen and calcium carbonate application rates on the was performed by using a Thermoquest gas chromatograph with a flame ionization detector (FID). The analysis was carried out using growth and essential oil content and composition of S. a fused silica capillary DB-5 column (30 m × 0.25 mm i.d.; film hortensis L. cv. Saturn cultivated in Iran. thickness 0.25 µm). The operating conditions were as follows: injector and detector temperatures were 250 and 280°C, respectively; nitrogen was used as the carrier gas at a flow rate of 1 MATERIALS AND METHODS ml/min; oven temperature was programmed from 60 -250°C at the rate of 4°C/min; and finally held isothermally for 10 min. The split ratio was 1/50. Gas chromatography-mass spectrometry (GC-MS) Field experiment analysis was performed by using a Thermoquest-Finnigan gas chromatograph equipped with above mentioned column and The experiment was conducted in 2008 at the experimental garden coupled to a TRACE mass quadrupole analyzer. The analysis was of the Research Center of Horticultural Sciences Department, carried out using a fused silica capillary DB-5 column (60 m × 0.25 University of Tehran, Karaj, Iran (altitude 1320 m, longitude 51 ْ E, mm i.d.; film thickness 0.25 µm). Temperature programming latitude 35 ْ 48 َ N). The average day and night temperatures during conditions were as given for GC. Helium was used as the carrier experiment were 26.93 and 20.75°C, respectively. At the beginning gas with ionization voltage of 70 eV. Ion source and interface of study, some physical and chemical characteristics of the soil at temperatures were 200 and 250°C, respectively. Mass range was the depth of 30 cm were analyzed (Table 1). The seeds of S. from m/z 43 - 456. hortensis cv. Saturn, a bred cultivar with high oil yield provided by Zardband Pharmaceutical Company located in Tehran, Iran, were sown in greenhouse on July, 5 in small pots (6 x 6 cm, 10 cm deep) Identification of compounds containing sand, clay and humus (1: 1: 1). After 4 weeks from sowing, the seedlings with 7 - 10 cm height were transplanted to The constituents of the oil were identified by calculation of their the field at a density of 6.35 plants/m 2 (0.45 × 0.35 m distance). The retention indices under temperature programmed conditions for Mumivand et al. 1861

Table 2. Chemical properties of soils with different levels of CaCO 3 after the harvest of S. hortensis cv. Saturn.

z Fe Av. Mn Av. Zn Av. Cu Av. Lime Organic C P Av. K Av. Total N CaCO 3 pH (mg/kg) (mg/kg) (mg/kg) (mg/kg) (%) (%) (mg/kg) (mg/kg) (%) (t/ha) 15.2 21 7.4 2.6 6.4 8.9 1.23 161 835 0.126 0 14.5 20.8 6 2.4 7.2 14.4 1.28 157 865 0.128 5 12.5 17.2 3 2.6 7.9 22 1.23 141 835 0.124 10

z: available.

-1 n-alkanes (C 6-C24 ) and the oil on a DB-5 column under the same with 5 t. ha CaCO 3 (200.23 and 46.68 g/plant, chromatographic conditions. Identification of individual compounds respectively) (Table 3). S. hortensis drug yield was was made by comparison of their mass spectra and retention significantly increased by higher levels of nitrogen (164.7 indices with those authentic samples and those given in the 2 -1 literature (Adams, 2001). Quantification of the relative amount of g/m at 150 Kg. ha ), whereas CaCO 3 treatments had no the individual components was performed according to the area significant effect on the drug yield (Table 3). percentage method without consideration of calibration factor. Leaf N content increased significantly with nitrogen fertilization as the highest value (4.4% in dried leaf) was observed with application of 150 Kg. ha -1. However, the Data analysis effect of N fertilizer on leaf Ca 2+ content was not 2+ Data of all measured parameters were subjected to variance significant. Leaf Ca content was significantly increased analysis using MSTAT-C statistical software. Also, Duncan’s by CaCO 3 treatment as the greatest percentage (2.07% in -1 multiple range tests was used to compare treatment means at a dried leaf) was observed in 10 t. ha CaCO 3. Also, the probability level of 0.05. CaCO 3 application had a significant negative effect on leaf N content as it was the highest (5.36% in dried leaf) -1 at 150 Kg. ha N when no CaCO 3 was applied (Table 4). RESULTS

Plant growth characters Essential oil content and yield

The results of the present study showed that nitrogen The effect of varying levels of soil nitrogen on the treatment significantly increased the plant height of S. essential oil content of plants was not considerable while hortensis (37.64 cm at 100 Kg. ha -1 compared to 35.25 the essential oil yield was significantly increased by N -1 cm at the control). However, the effect of CaCO 3 on this application. The highest essential oil yield (72.7 lit. ha ) parameter was not significant (Table 3). Width of plant was achieved through the application 150 Kg. ha -1 N. On was not significantly affected with nitrogen fertilization. the other hand, the essential oil content of summer -1 Nevertheless, CaCO 3 application had significant but savory was increased with CaCO 3 treatment up to 5 t. ha negative effect on the plant width as it was the highest at (4.518% v/w). A similar pattern was observed for the control treatment (39.8 cm) (Table 3). essential oil yield as the maximum oil content (71.27 lit. -1 -1 Effect of different N levels on stem diameter was ha ) was obtained by 5 t. ha of CaCO 3. The interaction significant and the maximal stem diameter was achieved between nitrogen and CaCO 3 applications was significant from 150 Kg. ha -1 (8.91 mm). On the contrary, it was not for the essential oil content, but not for the essential oil affected by CaCO 3 applications (Table 3). N fertilization yield (Table 4). significantly increased the mean of LA of plants, as the highest value was observed at 150 Kg. ha -1 (67.08 mm 2). Also, this parameter was significantly affected by CaCO3 Essential oil composition treatment and it was the greatest (64.9 mm 2) at the intermediate level of CaCO 3 (Table 3). The effects of nitrogen and calcium carbonate treatments Nitrogen fertilizer significantly affected the fresh weight on the essential oil constituents of S. hortensis L. cv. of herb as well as the dry weight. The fresh weight was Saturn are presented in Table 5. The major components greater in the plants fertilized with 100 Kg. ha -1 (198.03 g. of the essential oil were found to be carvacrol and γ- plant -1) than the control plants (171.11 g. plant -1), but no terpinene, which are similar to those reported earlier further increase was observed in the plants fertilized with (Svoboda and Greenaway, 2003; Zawislak, 2008). N 150 Kg/ha. Also, the highest value for dry weight (48.53 fertilizer had significant effect only on γ-terpinene -1 g. plant ) was achieved at 150 kg/ha. CaCO 3 treatment percentage of the oil. CaCO 3 significantly increased the altered the fresh and dry weights of herbs, significantly. relative percentages of major compounds such as The highest plant fresh and dry weights were obtained carvacrol and γ-terpinene and some minor components 1862 J. Med. Plant. Res.

Table 3. Effects of CaCO 3 and nitrogen treatments on the growth characteristics of S. hortensis L. cv. Saturn.

Plant Stem Means of fresh Drug Plant height Dry weight Treatment width diameter leaf area weight yield (cm) (g/plant) (cm) (mm) (mm 2) (g/plant) (g/m 2)

CaCO 3 (t/ha) 0 36.600 39.883 a 8.145 64.037 a 181.358 b 42.292 b 147.098 5 37.333 39.367 a 8.449 64.937 a 200.233 a 46.683 a 157.983 10 35.950 37.158 b 8.499 60.224 b 176.058 b 43.046 b 152.643 n.s. z n.s. n.s.

Nitrogen (Kg/ha) 0 35.256 b 38.244 8.078 b 59.261 b 171.111 b 39.586 b 135.996 b 50 36.389 ab 37.867 7.867 b 60.837 b 176.367 b 41.572 b 148.138 b 100 37.644 a 39.544 8.602 a 65.082 a 198.033 a 46.334 a 161.445 a 150 37.222 a 39.556 8.910 a 67.083 a 198.022 a 48.537 a 164.719 a n.s.

Interaction between CaCO 3 and N

CaCO 3 0, N 0 35.800 39.533 7.813 60.787 169.767 39.200 125.328

CaCO 3 0, N 50 36.000 38.067 7.393 63.243 169.000 37.950 136.821

CaCO 3 0, N 100 37.533 40.800 8.473 64.683 192.000 44.913 161.354

CaCO 3 0, N 150 37.067 41.133 8.900 67.433 194.667 47.107 164.888

CaCO 3 5, N 0 36.100 38.600 8.007 61.407 177.333 40.543 138.705

CaCO 3 5, N 50 36.567 39.267 8.167 62.350 188.867 43.000 149.627

CaCO 3 5, N 100 38.867 40.000 8.493 68.063 220.333 50.440 168.550

CaCO 3 5, N 150 37.800 39.600 8.130 67.927 214.400 52.750 175.048

CaCO 3 10, N 0 33.867 36.600 8.413 55.590 166.233 39.013 143.954

CaCO 3 10, N 50 36.600 36.267 8.043 56.917 171.233 43.767 157.967

CaCO 3 10, N 100 36.533 37.833 8.840 62.500 181.767 43.650 154.432

CaCO 3 10, N 150 36.800 37.933 8.700 65.890 185.000 45.753 154.220 n.s. n.s. n.s. n.s. n.s. n.s. n.s.

z: no significant effect.

such as β-bisabollene and α-pinene. The maximum carboxylase (Rubisco) (Dordas and Sioulas, 2008). percentage of carvacrol (47.2%) and γ-terpinene (41.3%) Where there is adequate N supply in soil, photosynthesis -1 were obtained from 10 and 5 t. ha of CaCO 3, rate increases and enables the plant to grow rapidly and respectively. This CaCO 3 effect was accompanied with a produce considerable biomass. In addition, application of decrease in α-terpinene, myrcene, p-cymene and α- nitrogen fertilizer increases the uptake and accumulation thujene percentages. In addition, the interaction between of other nutrients such as phosphorus (P) and potassium CaCO 3 and N treatments was significant for γ-terpinene, (K) (Baranauskiene et al., 2003). So, nitrogen plays an α-terpinene, myrcene and β-caryophyllene (Table 5). important role in vegetative growth and basic metabolism of the plant, which might be directly or indirectly involved in production of secondary metabolites (Baricevic and DISCUSSION Zupancic, 2002). Under conditions of nitrogen deficiency growth and development are inhibited and the plant yield The present study showed that nitrogen fertilization decrease due to the reduction of chlorophyll content and significantly influence the growth and performance of S. Rubisco activity (Dordas and Sioulas, 2008). High hortensis cv. Saturn, which is in agreement with the nitrogen fertilization may also cause a decrease in results of Ayanoglu et al. (2002), Baranauskiene et al. activities of phosphoenolpyrovate carboxylase (PEP) and (2003) and Sifola and Barbieri (2006). According to Rubisco enzymes thereby reduces the photosynthesis Ashraf et al. (2005) and Dordas and Sioulas (2008), the rate, plant growth and accumulation of secondary leaf N content was increased along with nitrogen products (Palada et al., 1995; Ashraf et al., 2006). application. Up to 75% of the leaf N content is present in Although, in this experiment, the effect of nitrogen chloroplasts mainly as a part of riboluse bisphosphate application on the oil content was not significant, Mumivand et al. 1863

2+ Table 4. Effects of CaCO 3 and nitrogen treatments on the essential oil content and yield and the Ca and N content of dried leaves of S. hortensis L. cv. Saturn.

Treatment Essential oil content (v/w) Essential oil yield (Lit/ha) Leaf N content (%) Leaf Ca 2+ content (%)

CaCO 3 (t/ha) 0 4.388 b 64.622 b 4.656 a 1.578 c 5 4.518 a 71.273 a 3.808 b 1.829 b 10 4.513 a 68.900 ab 3.617 b 2.072 a

Nitrogen (Kg/ha) 0 4.467 60.891 b 3.649 c 1.771 50 4.512 66.898 a 3.960 b 1.780 100 4.492 72.548 a 4.096 b 1.794 150 4.419 72.723 a 4.402 a 1.960 n.s. z n.s.

Interaction between CaCO 3 and N

CaCO 3 0, N 0 4.207 d 52.699 3.947 c 1.547

CaCO 3 0, N 50 4.460 abc 61.092 4.520 b 1.547

CaCO 3 0, N 100 4.467 abc 71.977 4.793 b 1.580

CaCO 3 0, N 150 4.417 bc 72.718 5.363 a 1.640

CaCO 3 5, N 0 4.577 ab 63.491 3.680 cd 1.730

CaCO 3 5, N 50 4.577 ab 68.488 3.717 cd 1.900

CaCO 3 5, N 100 4.560 ab 76.837 3.828 c 1.757

CaCO 3 5, N 150 4.357 cd 76.273 4.007 c 1.930

CaCO 3 10, N 0 4.617 a 66.481 3.320 d 2.037

CaCO 3 10, N 50 4.500 abc 71.114 3.643 cd 1.893

CaCO 3 10, N 100 4.450 abc 68.829 3.667 cd 2.047

CaCO 3 10, N 150 4.483 abc 69.176 3.837 c 2.310 n.s. n.s.

z: no significant effect.

essential oil production was increased due to higher levels of soil CaCO 3. Moreover, Quaggio et al. (2004) biomass yield. Such a relationship between nitrogen mentioned that the effect of CaCO 3 treatment on peanut levels and plant response has been observed in different yield was depended on many factors including the initial species such as Artemisia annua (Ayanoglu et al., 2002), soil Ca and Mo content, time of CaCO 3 application and Thymus vulgaris (Baranauskiene et al., 2003) and Nigella cultivar. The increase in the plant growth and yield up to -1 sativa (Ashraf et al., 2006). In agreement with the results 5 t. ha CaCO 3 could be explained by several reasons of Baranauskiene et al. (2003) on T. vulgaris , the including enhancement of exchangeable Ca and Mg, percentage of most of the oil constituents of S. hortensis increase of available Mo, decrease of Al, Mn and heavy was not significantly affected by nitrogen fertilization. metals toxicity due to increase of pH and improvement of As earlier expressed, the plant growth was increased soil characteristics (increase of pore volume and optimum -1 with application of CaCO 3 up to 5 t. ha while further moisture content) (Quaggio et al., 2004; Bakker et al., increase in CaCO 3 reduced the growth. Similar result has 2005; Rajasekaran, 2005). It seems that the decrease in -1 been reported by Supanjani et al. (2005a), as the plant the growth by application of 10 t. ha CaCO 3 could be growth and yield of Chrysanthemum coronarium L. was related to excessive Ca content of soil solution which -1 increased by applying of CaCO 3 up to 2 t. ha while causes to precipitation of P, S and Zn (Tunesi, 1999; higher amounts decreased yield. Also, Lee and Yang Wenming, 2001; Bakker et al., 2005; Rajasekaran, 2005). 2- (2005) reported an increase in the plant growth and yield Also, HPO 4 that is more easily adsorbed to soil matrix - of Ch. boreale with increasing of CaCO 3 levels up to 1.5 t. and taken up more slowly than H 2PO 4 by roots, is more ha -1. Since S. hortensis is characterized as a calcicole abundant on pH above 7.2 (Bakker et al., 2005). In our species and in natural habitats it is mainly found on study, chemical analysis of the soils treated with different calcareous soils (Ghahraman, 1996), this plant levels of CaCO 3 (Table 2) showed that the lime responded more positively than other species to higher percentage of soil increased to 22% in third level of 1864 J. Med. Plant. Res.

Table 5. Effects of CaCO 3 and N treatments on the essential oil constituents of S. hortensis L. cv. Saturn.

α-thujene α-pinene β-pinene Myrcene α-terpinen p-cymen γ-terpinen Thymol Carvacrol β-caryophyllen β-bisabollen Treatment (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%)

CaCO 3 (t/ha) 0 2.055 a 0.768 b 0.201 2.732 a 4.016 a 2.026 a 40.081 b 0.101 43.973 c 0.516 0.721 a 5 1.599 b 0.797 a 0.213 2.085 b 3.787 b 1.560 b 41.309 a 0.930 45.665 b 0.507 0.656 ab 10 1.061 c 0.772 ab 0.207 2.044 b 3.132 c 1.456 b 41.157 a 0.100 47.261 a 0.454 0.493 b n.s. z n.s. n.s. Nitrogen (Kg/ha) 0 1.548 0.774 0.207 2.384 3.611 1.347 40.129 b 0.107 45.917 0.542 0.720 50 1.590 0.787 0.210 2.324 3.691 1.781 41.461 a 0.105 45.266 0.466 0.521 100 1.558 0.770 0.203 2.224 3.589 1.648 40.327 ab 0.090 46.282 0.465 0.576 150 1.592 0.785 0.207 2.216 3.691 1.947 41.479 a 0.090 45.068 0.496 0.676 n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s.

Interaction between CaCO 3 and N

CaCO 3 0, N 0 1.982 0.757 0.196 3.017 a 3.643 d 1.487 38.330 c 0.128 44.180 0.670 a 0.963

CaCO 3 0, N 50 2.066 0.767 0.198 2.620 bc 4.073 ab 2.277 39.710 bc 0.100 44.967 0.490 bc 0.729

CaCO3 0, N 100 2.074 0.766 0.207 2.626 bc 4.128 ab 2.028 40.430 bc 0.080 43.987 0.395 c 0.545

CaCO 3 0, N 150 2.098 0.782 0.202 2.666 b 4.221 a 2.312 41.830 ab 0.095 42.760 0.508 bc 0.648

CaCO 3 5, N 0 1.578 0.781 0.205 1.809 f 3.982 abc 1.434 40.490 bc 0.088 46.787 0.589 ab 0.718

CaCO 3 5, N 50 1.621 0.814 0.226 2.191 de 3.813 bcd 1.515 42.920 a 0.099 43.837 0.495 bc 0.622

CaCO 3 5, N 100 1.576 0.794 0.210 2.132 de 3.626 d 1.440 40.500 bc 0.089 46.410 0.490 bc 0.694

CaCO 3 5, N 150 1.621 0.799 0.210 2.207 de 3.728 cd 1.852 41.320 ab 0.095 45.627 0.454 bc 0.589

CaCO 3 10, N 0 1.083 0.785 0.219 2.326 cd 3.207 e 1.120 41.560 ab 0.106 46.783 0.366 c 0.479

CaCO 3 10, N 50 1.082 0.779 0.206 2.162 de 3.186 e 1.552 41.740 ab 0.114 46.993 0.414 c 0.213

CaCO 3 10, N 100 1.024 0.750 0.193 1.913 ef 3.012 e 1.475 40.040 bc 0.101 48.450 0.509 bc 0.489

CaCO 3 10, N 150 1.057 0.774 0.208 1.774 f 3.124 e 1.676 41.270 ab 0.080 46.817 0.525 abc 0.791 n.s. n.s. n.s. n.s. n.s. n.s. n.s.

z: no significant effect.

-1 CaCO 3. Soil pH was increased from 6.4 (in in agreement with the results of Supanjani et al. higher levels (5 t. ha ) decreased. This contrast -1 control) to 7.9 (in 10 t. ha CaCO 3) , whereas (2005b), in which high concentration of Ca in could be due to the use of higher level of CaCO 3 exchangeable phosphorous and majority of nutrient solution remarkably decreased the N in our experiment and different soil conditions. micronutrients were decreased accompanied with content of leaves and flowers of Ch. coronariom . Also, the increase of Ca 2+ concentration in the CaCO 3 application. In another study, Lee and Yung (2005) reported leaves accompanied with CaCO 3 application has In this study, it was observed that leaf N content that N content of flower of Ch. boreale increased been observed in other studies (Lee and Yang, was decreased with application of CaCO 3. This is with application of low level of CaCO 3 while under 2005; Supanjani et al., 2005a). According to the Mumivand et al. 1865

results of Lee and Yung (2005) and Supanjani et al. fertilization under rain fed conditions. Ind. Crop. Prod., 27: 75-85. (2005a, b), CaCO treatment changed the amounts of Ghahraman A (1996). Color Atlas of Iranian Flora (In Persian). 3 Research Institute of Forests and Rangelands Publishing. major components of the essential oil and had significant Gulluce M, Sokmen M, Daferera D, Agar G, Ozkan H, Kartal N, effects on the oil content and yield. Increasing of Polissiou M, Sokmen A, Sahin F (2003). In vitro antibacterial, carvacrol percentage of summer savory essential oil was antifungal, and antioxidant activities of the essential oil and methanol accompanied with the decrease of percentages of extracts of herbal parts and callus cultures of S atureja hortensis . J. Agric. Food Chem., 51: 3958-3965. hydrocarbon procures such as p-cymene, α-thujene, Hadian J, Tabatabaei SMF, Naghavi MR, Jamzad Z, Ramak-Masoumi T myrcene and α-terpinene. This means that the rate of (2008). Genetic diversity of Iranian accessions of Satureja hortensis transformation of hydrocarbon precursors to carvacrol as L. based on horticultural traits and RAPD markers. Sci. Hortic., 115: a phenolic compound is higher in calcareous soils. The 196-202. Hajhashemi V, Sadraei H, Ghannadi AR, Mohseni S (2000). same phenomenon has been reported on T. vulgaris and Antispasmodic and antidiarrhoeal effect of Satureja hortensis L. T. piperella , as the populations growing on calcareous essential oil. J. Ethnopharmacol. 71: 1-2. soils contain higher carvacrol content (Boira and Lee KD, Yang MS (2005). Changes in mineral and terpene Blanquer, 1998; Thompson et al., 2003). concentration following calcium fertilization of Chrysanthemum boreale M. Res. J. Agric. Biol. Sci., 1: 222-226. Martins FT, Santos MH, Polo M, Barbosa LCA (2007). Effects of the interactions among macronutrients, plant age and photoperiod in the Conclusion composition of Hyptis suaveolens (L.) Poit essential oil from Alfenas (MG), Brazil. Flavour Fragr. J., 22: 123-129. Ozcan M (2004). Mineral contents of some plants used as condiments The economic value of summer savory for cultivation as a in Turkey. Food Chem., 84: 437-440. cash crop depends on the drug and oil yield as well as Palada MC, Crossman SMA, Kowalski JA (1995). Growth and yield the concentration of carvacrol and γ-terpinene in the oil. response of Thyme ( Thymus vulgaris L.) to sources of nitrogen N fertilization had positive effects on the drug and oil fertilizer. In: Palada, C., Clarke, C. C. (Eds.), Caribbean Economic Stabilization through Agricultural Development Strategies. The yield. The optimum level of nitrogen fertilizer for this crop University of the Virgin Islands Cooperative Extension Service and -1 was 100 Kg. ha . Based on the obtained results, CaCO 3 the Carribbean Food Crops Society, Virgin Islands, pp. 58-64. has the capacity to alter the relative percentages of S. Pharmacopee Europeene (1997). 3 Ed., Maisonneuve, Sante Ruffine. hortensis essential oil main components. Also, Prohens J, Rodriguez-Burruezo A, Nuez F (2003). New crops: an -1 alternative for the development of horticulture. J. Food, Agric. application of 5 t. ha CaCO 3 gave the highest essential Environ., 1: 75-79. oil content and yield with high relative percentages of Quaggio JA, Gallo PB, Owino-Gerroh C, Abreu MF, Cantarella H carvacrol and γ-terpinene. In conclusion, application of (2004). Peanut response to lime and molybdenum application in low 100 Kg. ha -1 N with t. ha -1 CaCO was identified as the pH soils. Rev. Bras. Cienc. Solo., 28: 659-664. 3 Rajasekaran G (2005). Sulphate attack and ettringite formation in the best treatment for the production of essential oil. lime and cement stabilized marine clays. Ocean Eng., 32: 1133-1159. Rechinger KH (1982). Satureja . Flora Desiranischen Hoclandes and der Umrahmenden Gebirge, vol. 150. Akademische Druku Verlags Antalt REFERENCES Graz, Austria, pp. 495-504. Sahin F, Karaman I, Gulluce M, Oguteu H, Sengul M, Adıguzel A, Adams RP (2001). Identification of essential oil components by gas Ozturk S, Kotan R (2003). Evaluation of antimicrobial activities of chromatography/quadrupole mass spectroscopy. Allured Publishing Satureja hortensis L. J. Ethnopharmacol., 87: 61-65. Corporation, Carol Stream, IL. Salmasi SZ, Heidari F, Alyari H (2008). Effect of microelements and AOAC (1990a). Metals in Plants. In: Helrich, K. (Ed.), Official Methods plant density on biomass and essential oil production of peppermint of Analysis. AOAC Inc., Arlington, VA, p. 42. (Mentha piperita L.). plant Sci. Res., 1: 24-26. AOAC (1990b). Nitrogen (total) (crude proteins) in Plants. In: Helrich, K. Sefidkon F, Abbasi K, Khaniki GB (2006). Influence of drying and (Ed.), Official Methods of Analysis. AOAC Inc., Arlington, VA, p. 59. extraction methods on yield and chemical composition of the Ashraf M, Ali Q, Iqbal Z (2006). Effect of nitrogen application rate on the essential oil of Satureja hortensis. Food Chem., 99: 19-23. content and composition of oil, essential oil and minerals in black Sifola MI, Barbieri G (2006). Growth, yield and essential oil content of cumin ( Nigella sativa L.) seeds. J. Sci. Food Agric., 86: 871-876. three cultivars of basil grown under different levels of nitrogen in the Ashraf M, Ali Q, Rha ES (2005). The effect of applied nitrogen on the field. Sci. Hortic., 108: 408-413. growth and nutrient concentration of Kalonji ( Nigella sativa ). Aust. J. Skocibusic M, Bezic N, Dunkic V (2006). Phytochemical composition Exp. Agric., 45: 459-463. and antimicrobial activities of the essential oils from Satureja Ayanoglu F, Mert A, Kirici S (2002). The effects of different nitrogen subspicata Vis. growing in Croatia. Food Chem., 96: 20-28. doses on Artemisia annua L. J. Herbs Spi. Med. Plants, 9: 399-404. Suh EJ, Park KW (2000). Effect of calcium ion in nutrient solution on the Bakker C, Rodenburg J, Bodegom P (2005). Effects of Ca- and Fe-rich content and composition of essential oil of sweet basil in hydroponics. seepage on P availability and plant performance in calcareous dune J. Kor. Soc. Hortic. Sci., 41: 598-601. soils. Plant Soil, 275: 111-122. Supanjani, Tawaha ARM, Yang MS, Han HS, Lee KD (2005a). Role of Baranauskiene R, Venskutonis PR, Viskelis P, Dambrauskiene E calcium in yield and medicinal quality of Chrysanthemum Coronarium (2003). Influence of nitrogen fertilizers on the Yield and Composition L. J. Agron., 4: 186-190. of Thyme ( Thymus vulgaris ). J. Agric. Food Chem., 51: 7751-7758. Supanjani, Tawaha ARM, Yang MS, Han HS, Lee KD (2005b). Calcium Baricevic D, Zupancic A (2002). The impact of drought stress and/or effects on yield, mineral uptake and terpene components of nitrogen fertilization in some medicinal plants. J. Herbs Spi. Med. hydroponic Chrysanthemum coronarium L. Res. J. Agric. Biol. Sci., 1: Plants, 9: 53-64. 146-151. Boira H, Blanquer A (1998). Environmental factors affecting chemical Svoboda KP, Greenaway RI (2003). Investigation of volatile oil glands variability of essential oils in Thymus piperella L. Biochem. Syst. of Satureja hortensis L. (summer savory) and phytochemical Ecol., 26: 811-822. comparison of different varieties. J. Aromather., 13: 196-202. Dordas CA, Sioulas C (2008). Safflower yield, chlorophyll content, Thompson JD, Chalchat JC, Michet A, Linhart YB, Ehlers B (2003). photosynthesis, and water use efficiency response to nitrogen Qualitative and quantitative variation in monoterpene co-occurrence 1866 J. Med. Plant. Res.

and composition in the essential oil of Thymus vulgaris chemotypes. Zargari A (1970). Medicinal Plants (In Persian), vol. 4. Tehran J. Chem. Ecol., 29: 859-880. University, pp. 42-45. Tunesi S, Poggi V, Gessa C (1999). Phosphate adsorption and Zawislak G (2008). Yielding and quality of herb of garden savory precipitation in calcareous soils: the role of calcium ions in solution (Satureja hortensis L.) cv. Saturn grown from transplants (In Polish, and carbonate minerals. Nutr. Cycl. Agroecosyst. 53: 219-227. with English abstract). Zeszyty Problemowe Postepow Nauk Wenming D, Zhijun G, Jinzhou D, Liying Z, Zuyi T (2001). Sorption Rolniczych. 527: 327-333. characteristics of zinc(II) by calcareous soil radiotracer study. Appl. Radiat. Isot., 54: 371-375. Yang MS, Tawaha ARM, Lee KD (2005). Effects of ammonium concentration on the yield, mineral content and active terpene components of Chrysanthemum Coronarium L. in a hydroponic system. Res. J. Agric. Biol. Sci., 2: 170-175.