See

discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/288838690

Response of damask rose (Rosa damascena

Mill.) to foliar application of magnesium (Mg),
copper (Cu) and zinc (Zn) sulphate under
Western Himalayas.

Article in Industrial Crops and Products · December 2015

DOI: 10.2016/j.indcrop.2015.12.065.

CITATIONS READS

0 111

5 authors, including:

Rakesh Kumar Saurabh Sharma

CSIR - Institute of Himalayan Bioresource T… CSIR - Institute of Himalayan Bioresource T…
74 PUBLICATIONS 185 CITATIONS 36 PUBLICATIONS 112 CITATIONS

SEE PROFILE SEE PROFILE

Munish Kaundal
CSIR - Institute of Himalayan Bioresource T…
4 PUBLICATIONS 0 CITATIONS

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Protected cultivation of vegetables in the mountain regions of Himachal Pradesh View

project
All in-text references underlined in blue are linked to publications on ResearchGate, Available from: Munish Kaundal
letting you access and read them immediately. Retrieved on: 09 October 2016
 Industrial Crops and Products 83 (2016) 596–602

Contents lists available at ScienceDirect

Industrial Crops and Products

journal homepage: www.elsevier.com/locate/indcrop

Response of damask rose (Rosa damascena Mill.) to foliar application

of magnesium (Mg), copper (Cu) and zinc (Zn) sulphate under
western Himalayas
Rakesh Kumar a,∗ , Saurabh Sharma a,b , Munish Kaundal b , Shivani Sharma a ,
Meenakshi Thakur a
a
Agrotechnology and Plant Improvement Department, CSIR—Institute of Himalayan Bioresource Technology, (Council of Scientific and Industrial Research),
Post Box No. 6, Palampur, 176 061 HP, India
b
Academy of Scientific and Innovative Research, CSIR—Institute of Himalayan Bioresource Technology, (Council of Scientific and Industrial Research), Post
Box No. 6, Palampur, 176 061 HP, India

a r t i c l e i n f o a b s t r a c t

Article history: Foliar application of nutrients plays a vital role in plant metabolism. Very little research has been carried
Received 28 September 2015 out with foliar spray of nutrients on growth, yield, essential oil content and composition of damask rose
Received in revised form (Rosa damascena Mill.) under western Himalayas, hence the experiments were conducted from December
23 December 2015
2012 to May 2014, to study the effect of foliar application of MgSO4 , CuSO4 and ZnSO4 alone and in com-
Accepted 23 December 2015
binations at different concentrations for the first time. Eleven treatments, either single or mixture of
Available online 6 January 2016
salts were applied through foliar spray to damask rose plants according to experimental layout. Results
of the study revealed that foliar application of ZnSO4 @ 2% recorded 21.9% and 21.3% higher numbers of
Keywords:
Rosa damascena
flower/plant and flower yield/plant than control, respectively. Individual flower weight, essential oil con-
Hydro distillation tent, chlorophyll content index and number of branches/plant were not affected by different treatments.
Citronellol However, essential oil composition was significantly affected by foliar application of different nutrients.
E-geraniol Total 25 compounds were identified which accounted for 92.73 ± 0.8% to 95.64 ± 0.8% in different treat-
Nutrients ments. Application of MgSO4 @ 1% + ZnSO4 @ 1% recorded 26.4% higher citronellol + nerol (37.79 ± 1.4%)
Foliar fertilization content than control. Essential oil produced from the flowers in which plants were treated with ZnSo4
@ 1% recorded significantly higher composition of Z-rose oxide (2.25%), E-geraniol (26.53%) and lower
composition of nonadecene (1.53%), nonadecane (6.81%), docosane (0.58%) and heneicosane (2.60%) as
compared with other treatments.
© 2015 Elsevier B.V. All rights reserved.

1. Introduction factors, environmental factors (Danyaie et al., 2011), pre harvest

Rosa damascena Mill. (family Rosaceae) is an extensively pop-
ular oil-yielding crop and is being cultivated throughout because
of its industrial use in perfumery, flavoring, cosmetics, pharmaceu-
tical and aromatic industries and also because of its ornamental
value. It has cooling, soothing, astringent, anti-inflammatory and
cardiotonic effect (Yassa et al., 2009; Baniasad et al., 2015). R.
damascena commonly called as “Damask Rose” is predominantly
cultivated in Turkey, Bulgaria, Morocco, Iran, India, South Russia,
Southern France and China (Kazaz et al., 2010). A number of factors
are responsible for enhanced flower yield; these are usually genetic
∗ Corresponding author. Fax: + 91 1894 230433.
E-mail address: rakeshkumar@ihbt.res.in (R. Kumar).
factors such as soil fertility and nutrient availability, cultural prac-
tices, harvesting techniques, handling and post harvest factors
(Kumar et al., 2013a,b). Flower yield determines crop productiv-
ity in turn (Misra et al., 2005) which further adds to oil yield. Rose
oil is one of the most highly- priced oils among other natural oils
(Shamspur et al., 2012) because of low oil content in petals and lack
of natural and synthetic substitutes (Yilmaz et al., 2011). Damask
rose oil contains high percentage of acyclic monoterpene alcohols
exhibiting citronellol, E-geraniol and nerol in particular as well as
the aromatic alcohol phenylethyl alcohol and hydrocarbons such as
nonadecene, nonadecane and heneicosane (Mohamadi et al., 2011).
Macro and micronutrients play a significant role in the pro-
duction of high yield with high quality products (Sawan et al.,
2001). Though micronutrients are needed in small quantities; how-
ever, their small deficiencies can cause significant disturbance

http://dx.doi.org/10.1016/j.indcrop.2015.12.065
0926-6690/© 2015 Elsevier B.V. All rights reserved.
 R. Kumar et al. / Industrial Crops and Products 83 (2016) 596–602
in the physiological and metabolic processes in the plant (Bacha
et al., 1997). Magnesium helps strengthen cell walls and improves
plants’ uptake of nitrogen, phosphorus, and sulfur. Magnesium is an
important co-enzyme in many of the reactions of photosynthesis.
Magnesium is applied as MgSO4 and plants treated with it pro-
duce more new canes at the bottom of the plant (bottom breaks)
and darker green foliage in roses. Mg in chlorophyll accounts for
15–20% of the total Mg content of plants. In micronutrients, Cop-
per (Cu) is an essential microelement in higher plants as it occurs
as a part of the prosthetic groups of several enzymes. It was shown
to be associated with proteins or nuclear contaminants. Cu is an
important part of the protein, plastocyanin. Zinc (Zn) is an essen-
tial element for plant that act as a metal component of various
enzymes or as a functional structural or regulatory cofactor and
for protein synthesis, photosynthesis, the synthesis of auxin, cell
division, maintains membrane structure and function and sexual
fertilization (Marchner, 1995).
Movement of elements is usually faster through the stomata,
but the total absorption may be as great through the epidermis. Mg,
Cu and Zn are partially mobile essential nutrients. These nutrients
when applied through foliar application have recorded significantly
higher yield. Akhtar et al. (2009) have reported 28.2% higher essen-
tial oil of Mentha piperita by application of 3 ppm zinc chloride
foliar application compared with the control. Similarly, essential
oil biosynthesis in basil (Ocimum sanctum) was significantly influ-
enced by Fe and Zn application (Misra et al., 2006). Zinc application
stimulated the fresh and dry matter production, essential oil and
menthol concentration of Japanese mint (Misra and Sharma, 1991).
Nahed Abd El-Aziz and Balbaa (2007) reported that foliar spray-
ing with zinc (100 ppm) in blue sage (Salvia farinacea L.) enhanced
the length of peduncle, length of main inflorescence, number of
inflorescence and florets, and fresh and dry weight of inflores-
cences/plant. Micronutrients such as iron, manganese and zinc
have important roles in plant growth and yield of aromatic and
medicinal plants (Abd El- Wahab, 2008). The leaf total chlorophyll
contents, vase life and the mineral contents (NPK) of leaves were
significantly increased as a result of foliar application of micronu-
trients (Younis et al., 2013); they further concluded that application
of micronutrients along with NPK could improve flower yield and
quality of roses. Ahmad et al. (2010) reported that plants with-
out micronutrients application produced poor quality vegetative
growth and less number of flowers; further concluded that appli-
cation of micronutrients could help better to improve flower yield
and quality of roses. Foliar fertilization is an effective practice for
the application of some micronutrients, since it uses low rates
and the micronutrient does not directly contact the soil, avoiding
losses through fixation (Volkweiss, 1991). Foliar spray of different
micronutrients has also been reported to be equally or more effec-
tive as soil application by different researchers (Torun et al., 2001).
Influence of Mg, Cu, ZnSO4 on the yield and essential oil of damask
rose plants under western Himalayas is not documented. Therefore,
the present field experiment was conducted to study in detail the
effect of foliar application of Mg, Cu and ZnSO4 on yield attributes,
biomass yield, essential oil yield, major essential oil constituents of
R. damascena under western Himalayas for the first time.
2. Materials and methods
2.1. Experimental site
The present study was conducted in the experimental farm
of CSIR—Institute of Himalayan Bioresource Technology (Coun-
cil of Scientific and Industrial Research), Palampur, India located
at 32◦ 06 05 N latitude, 76◦ 34 10 E longitude and an altitude of
1325 m above the mean sea level during 2012 and 2014. The site
597
experiences mean maximum temperature from 18.7 to 31.5 ◦ C dur-
ing 2013 and 17.2 to 30.4 ◦ C during 2014 in bud formation and
flowering period. Minimum temperature ranged from 7.5 to 18.6 ◦ C
and 6.8 to 17.7 ◦ C during 2013 and 2014, respectively (Table 1). Rel-
ative humidity during flowering season ranged from 41.1 to 72.1%
and 50.7 to 62.6% during 2013 and 2014, respectively.
2.2. Experimental details
The experiment was laid out in randomized block design (RBD)
with three replications. Soil of the experimental plot was sandy
loam in texture (11.6% sand, 37.6% silt and 50.8% clay) acidic (pH
5.7), having available N 0.548 ppm, available P 0.094 ppm, avail-
able K 0.578 ppm, available Mg 140.6 ppm, available Cu 3.5 ppm
and available Zn 6.1 ppm (Table 2). The experiment consisted of
eleven treatments viz., T1: Control (water spray); T2: MgSO4 (1%);
T3: MgSO4 (2%); T4: CuSO4 (1%); T5: CuSO4 (2%); T6: ZnSO4 (1%); T7:
ZnSO4 (2%); T8: MgSO4 (1%) + CuSO4 (1%); T9: MgSO4 (1%) + ZnSO4
(1%); T10: CuSO4 (1%) + ZnSO4 (1%); T11: MgSO4 (1%) + CuSO4
(1%) + ZnSO4 (1%). Foliar application of nutrients was done two
times, at first when plants had 6 or 8 leaves and second time was
applied when these had 10 or 12 leaves (early of flowering stage).
Mg was applied as MgSO4 ·7H2 0 containing 10% of Mg. Cu was
applied as CuSO4 and Zn was applied as ZnSO4 . For the foliar appli-
cation, knap sac sprayer was used at 250 kPa pressure. To avoid
the effect of micronutrient spraying, the plots were separated by
borders of 1.5 m in width from all sides. The damask rose acces-
sion Jwala (no. PLP1650711) (Kumar et al., 2013c) was used for this
experiment. All the experimental plots (10 treatments × 3 replica-
tions = 30 plots of 135 m2 size with 120 rose bushes in each plot)
received 90 kg N, 60 kg P and 90 kg K per hectare as urea, single
super phosphate and muriate of potash, respectively. N was applied
in two equal splits, one after hoeing in the month of December
2012 and December 2013 as top dressing. The first dose of N was
applied along with P and K after hoeing in the month of December
every year and the second dose after 1 month later in January. The
plots were manually weeded to keep them weed free throughout
the experiment. Flood irrigations (2.5 cm depth) were given to the
crop at 10–15 days intervals to ensure adequate soil moisture for
good crop growth during the experimental period. Yield attributes
such as plant height, number of flowers/plant, flower yield per
plant, chlorophyll content index (CCI), number of branches/plant
were measured on ten randomly selected plants in each plot. CCI
was measured by chlorophyll concentration meter CCM 200 (Opti
sciences, Tyngsboro, USA). From each treatment five plants were
selected and from each plant three readings were measured from
third terminal leaf from upper side with the CCM-200 portable
chlorophyll meter (Opti-Sciences, Tyngsboro, MA), which calcu-
lates a unitless CCI value from the ratio of optical absorbance at
653 nm to that at 931 nm. Major veins and areas of obvious visual
damage or disease were avoided (Van den Berg and Perkins, 2004).
2.3. Essential oil extraction
Flowers were harvested during April & May months of 2013
and 2014. In each treatment 1000 g flowers were harvested. These
were hydro-distilled with 3 L of water in the 5 L capacity Cle-
venger type apparatus for 5 h. Flowers were harvested at 6:00 am
and were immediately brought to laboratory for distillation within
10 min after harvest. The quantities of rose oils obtained at the
end of distillation were measured as millilitre and % ratios (v/w%)
were determined according to standard procedure described in
European Pharmacopoeia (1975). All the essential oil samples were
dried over anhydrous sodium sulphate, and were stored at 4 ◦ C until
598 R. Kumar et al. / Industrial Crops and Products 83 (2016) 596–602

Table 1
Meteorological data during the crop growth.

Phenological stage MSW Maximum Minimum Rainfall Relative Wind Evaporation Sunshine
temp. (◦ C) temp. (◦ C) (mm) humidity speed (mm) hours (h)
(%) (km h−1 )

2013 M E
Bud 10 23.9 11.1 0.0 63.7 43.7 6.3 3.3 8.7
for- 11 21.4 8.9 9.4 67.1 49.7 6.9 2.9 5.6
ma- 12 18.7 7.5 174.6 72.1 57.9 6.4 2.3 6.2
tion 13 22.1 10.4 36.2 66.3 68.7 6.3 2.2 6.8
14 24.5 12.2 0.0 62.1 39.7 6.4 4.3 8.7
Flowering 15 26.9 13.4 23.0 64.1 40.6 6.5 4.4 8.9
16 26.1 14.0 18.6 59.9 43.7 7.0 4.3 7.4
17 25.1 14.9 13.0 67.9 48.9 6.1 3.9 6.1
18 29.7 17.1 0.0 46.3 26.9 7.1 5.8 8.1
19 30.4 16.6 27.2 41.1 44.0 7.3 6.1 7.1
20 31.5 18.6 0.0 42.7 27.0 6.7 7.9 10.4
2014
Bud 10 17.2 6.8 15.2 65.7 62.7 7.1 2.0 4.4
for- 11 21.3 9.1 39.4 64.7 48.1 6.1 2.2 5.0
ma- 12 20.5 9.2 63.6 63.0 52.0 7.0 2.7 4.6
tion 13 21.7 10.8 8.8 58.4 49.3 6.6 2.8 6.6
14 23.1 11.4 6.3 59.4 63.0 6.9 2.9 6.0
Flowering 15 23.8 11.4 0.4 52.0 34.0 6.9 3.8 8.6
16 22.8 11.6 49.3 62.6 45.3 6.3 3.1 5.9
17 27.8 15.0 0.6 50.7 35.3 6.1 5.2 10.8
18 30.4 17.7 7.6 59.1 41.0 6.4 4.9 9.6
19 27.9 15.3 38.0 62.0 47.7 6.3 3.7 6.4
20 26.9 14.6 0.0 61.1 42.3 5.8 5.0 8.4

MSW: Meteorological standard weeks, M: Morning; E: Evening.

analyzed by GC and GC–MS analysis. GC and GC–MS analysis were Table 2

Physico chemical properties of soil (0–30 cm soil depth).
carried out during June 2014.
Soil properties Values
2.4. Gas chromatography analysis Soil EC (dS/m) 0.072 ± 0.02
Soil pH 4.56 ± 0.29
Gas chromatographic analysis was carried out on a Shimadzu Organic carbon (%) 0.739 ± 0.31
Available N ppm 0.548 ± 0.06
GC-2010 gas chromatograph fitted with FID detector and a DB- Available P ppm 0.094 ± 0.01
5 capillary column (SGE International, Ringwood, Australia) of Available K ppm 0.578 ± 0.09
30 m length, 0.25 mm i.d. and 0.25 ␮m film thickness, 5% diphenyl, Available Mg ppm 140.6 ± 1.50
95% methyl polysiloxane stationary phase. Auto injection split Available Cu ppm 3.5 ± 0.11
Available Zn ppm 6.1 ± 0.71
mode was used with 2 ␮L injection volume after dissolving oil in
Soil texture Sandy loam
dichloromethane (dilution: 10 ␮L oil in 2 mL, GC grade). The oper- Sand (%) 11.6 ± 1.14
ating conditions were as follows: carrier gas nitrogen (99.9% pure) Silt (%) 37.6 ± 1.68
with a flow rate of 1.05 mL min−1 , the oven temperature was pro- Clay (%) 50.8 ± 1.73
grammed as follows: 70 ◦ C (4 min) and then 220 ◦ C at the rate of Data represent means ± standard deviation of triplicate samples.
4 ◦ C min−1 , injector and detector temperatures were set at 240 ◦ C
and 260 ◦ C, respectively.
puter library namely Wiley, New York mass spectral (MS) library,
National Institute of Standards and Technology, NIST (Stein, 2005).
2.5. Gas chromatography–mass spectrometry

Gas chromatography–mass spectrometry analysis was carried
out by GC–MS (QP2010 Shimadzu, Tokyo, Japan) equipped with
AOC-5000 Auto injector and DB-5 capillary column (SGE Inter-
national, Ringwood, Australia) of 30 m length, 0.25 mm i.d. and
0.25 ␮m film thickness. Temperature was programmed from 70 ◦ C
for 4 min and then increased at the rate of 4 ◦ C min−1 to 220 ◦ C and
held for 5 min. Mass spectrometer injector temperature 240 ◦ C and
interface temperature 250 ◦ C; sample injection volume 2 ␮L; split
ratio 1:50 and mass scan 50–800 amu. Helium (99.9% pure) was
used as a carrier gas with flow rate of 1.1 mL min−1 .
2.6. Identification of components
The retention index was calculated for all volatile constituents
by using homologous series of n-alkanes (C8 –C24 ) manufactured
by SUPELCO Sigma–Aldrich. The components of oil were identi-
fied by matching their retention indices (RI) and mass spectra with
those of reported in literature (Adams, 1995) and stored in the com-
2.7. Soil and plant analysis
Soil and plant samples were sampled from each plot after flower
harvest and dried at 70 ◦ C in forced air oven for 3 days and ground to
pass a 30 mesh screen. The available N was estimated by distilling
the soil (2 g) with alkaline potassium permanganate solution and
determining the ammonia liberated (Subbiah and Asija, 1956). Esti-
mation of available P and available K was done by method Mehlich
No. 3 (Mehlich, 1984). The organic matter content was estimated
by spectrophotometer HACH DR 2000 (Anonymous, 2000). Soil tex-
tural class was determined by hydrometer method and soil pH
was measured in a 1:2 soil:water suspension by glass electrode
pH meter (Black, 1965).
2.8. Heavy metal analysis
Analysis of heavy metals (Copper, Magnesium and Zinc) in soil
samples was performed as follows: Soil sample (1 g) was digested in
 R. Kumar et al. / Industrial Crops and Products 83 (2016) 596–602 599

Table 3
Effect of foliar application of Mg, Cu and Zn sulphate on yield attributes, flower yield and essential oil of damask rose (Pooled data of two years).

Treatment Number of flowers plant−1 Flower yield plant−1 Flower weight (g) Oil content (%) CCI Number of branches
e
T1: Control 107.8 451.2d 4.9 0.043 17.5 17.0
T2: MgSO4 (1%) 115.9cd 455.3d 4.3 0.052 17.8 15.7
T3: MgSO4 (2%) 120.5bcd 453.6d 4.2 0.046 19.0 17.1
T4: CuSO4 (1%) 121.8bcd 464.2cd 4.0 0.045 18.7 16.5
T5: CuSO4 (2%) 138.5a 532.8ab 4.2 0.044 18.6 14.5
T6: ZnSO4 (1%) 114.3d 458.7d 4.4 0.046 19.3 15.1
T7: ZnSO4 (2%) 131.4ab 547.1a 4.9 0.043 17.6 16.4
T8: MgSO4 (1%) +CuSO4 (1%) 120.3cd 505.5abc 5.0 0.044 20.3 17.6
T9: MgSO4 (1%) +ZnSO4 (1%) 119.4bcd 531.6abc 5.2 0.046 19.3 15.7
T10: CuSO4 (1%) +ZnSO4 (1%) 123.0bcd 475.8bcd 4.2 0.044 18.2 16.3
T11: MgSO4 (1%) +CuSO4 (1%) +ZnSO4 (1%) 128.9abc 522.7abc 4.7 0.046 19.7 13.7

Alphabets followed by the same letter in a row are not significantly different.

glass tube of 250 mL along with 15 mL of nitric acid (HNO3 ) at 140 ◦ C
for 2 h. The content was allowed to cool down at room tempera-
ture and was evaporated to dryness. The dried sample was further
treated with 3 mL of HClO4 and was kept in fume hood for fur-
ther oxidation from the sample solution for half an hour at 240 ◦ C.
After digestion of the sample and cooling it down 10 mL of distilled
water was added to dilute it. The whole content was filtered and dis-
tilled water was added to make the final volume upto 100 mL. The
heavy metal analysis was performed with Shimadzu model AA 6300
Atomic Absorption Spectrophotometer (Tokyo) Japan by using dif-
ferent metal Hallow cathode lamps (HAMA—MATSU PHOTONICS
K.K. JAPAN) as radiation source.
2.9. Statistical analysis
Statistical analysis of data was done as per the standard anal-
ysis of variance (ANOVA) using the software SYSTAT-12 (SYSTAT
Software Inc., Chicago, IL, USA) and technique as applicable for the
randomized complete block design (RBD). Variance (F) ratio was
applied to test the significance of treatment variance versus error
variance. Least significant difference (LSD) values at 5% probabil-
ity (P ≤ 0.05) level were computed by multiplying standard error
of difference (SEd) values with tabulated t values, for assessing the
significance of differences between any two treatment means. In
case of significant treatment effects, a comparison of means was
performed by means of Duncan’s multiple range test method at a
significance level of 5 percent (P ≤ 0.05).
3. Results and discussion
3.1. Yield attributes and yield
Number of flowers/plant is one of the main yield attributing
characters of damask rose. Flowers per plant were significantly
affected by foliar application of nutrients and these ranged from
107.8 to 138.5 flowers/plant in different treatments (Table 3).
Foliar application of CuSO4 @ 2% (T5) recorded significantly higher
flowers/plant (138.5) as compared with other treatments but
remained at par with treatment T7 (ZnSO4 @ 2%) and T11 (MgSO4
@ 1% + CuSO4 @ 1% + ZnSO4 @ 1%). Treatment T5 (CuSO4 @ 2%)
recorded 28.5% higher flowers per plant than control. Likewise
flower number/plant, flower yield/plant was also significantly
affected by foliar application of nutrients (Table 3). Foliar feeding of
ZnSO4 @ 2% (T7) produced significantly higher flower yield/plant
as compared with other treatments and control but remained at
par with treatment T5 (CuSO4 @ 2%); T9 (MgSO4 @ 1% + ZnSO4 @
1%), T11 (MgSO4 @ 1% + CuSO4 @ 1% + ZnSO4 @ 1%) and T8 (MgSO4
@ 1% + CuSO4 @ 1%). Foliar application of ZnSO4 @ 2% (T7) recorded
21.3% higher flower yield/plant as compared with control. A similar
effect of Zn supply on flower yield was also reported on Matricaria
chamomilla (Grejtovsky et al., 2006; Nasiri et al., 2010), Coriandrum
sativum (Said-Al Ahl and Omer, 2009) and Ocimum basilicum (Said-
Al Ahl and Mahmoud, 2010). Zinc is one of the eight essential trace
elements which is necessary for the normal healthy growth and
reproduction of crop plants (Parker et al., 1992). Zinc plays impor-
tant function in plant growth and yield of aromatic and medicinal
plants (Abd El-Wahab, 2008). Rao and Rajput (2011) observed that
Mg and micronutrients enhanced the total biomass yields of pal-
marosa (Cymbopogon martinii) by 37.0–42.6% and the total essential
oil yields by 44.6–81.7% in comparison to the control (water spray).
Flower weight was not affected by foliar application of differ-
ent nutrients (Table 3). Though combined application of MgSO4 @
1% + ZnSO4 @ 1% (T9) recorded heavier flowers than other treat-
ments. Similarly number of branches, CCI, fresh flower yield and
essential oil yield was also not affected by the foliar application of
different nutrients as compared with control. This may be due to
the fact that our soils are not deficient in Mg, Cu and Zn (Table 2).
3.2. Essential oil content and composition
Essential oil ranged from 0.043 to 0.052% in different treatments
(Table 3). Damask rose essential oil content was not significantly
affected by foliar application of nutrients. This may be due to
higher concentration of these nutrients in soil (Table 2). Whereas,
essential oil of M. piperita increased by 28.2% by foliar application
of 3 ppm zinc chloride compared with the control (Akhtar et al.,
2009). Likewise, Oregano essential oil yield increased (31%) by foliar
application of calcium and magnesium compared with the control
(Dordas, 2009). Misra and Sharma (1991) reported that Zn appli-
cation stimulated the fresh and dry matter production, essential
oil and menthol concentration of Japanese mint. Application of a
mixture of iron + zinc gave the highest essential oil of basil under
soil salinity condition in Egypt (Said-Al Ahl and Mahmoud, 2010).
Spraying micronutrients increased essential oils in plants viz. O.
sanctum (Youssef et al., 2004), thyme (Hanan Ali, 2007; Yadegari,
2012), chamomile (Nasiri et al., 2010), Rosa hybrida (Younis et al.,
2013). However, Shiriyan et al. (2014) did not find any significant
effect of foliar application of Fe and Zn alone or combinations on
yield of savory (Satureja hortensis) in silty clay loam soil (pH 7.6) in
Iran.
The analysis of the essential oils of damask rose led to the iden-
tification of 25 constituents (Table 4). Comparison of the analytical
data of the oils revealed marked differences in qualitative and
quantitative composition. The major constituents of the essential
oil were citronellol + nerol (29.89 ± 1.9 to 37.79 ± 1.4%), E-geraniol
(22.88 ± 0.9 to 27.82 ± 4.4%), phenyl ethyl alcohol (0.39 ± 0.1 to
1.01 ± 0.3%), Z-rose oxide (0.17 ± 0.0 to 1.31 ± 1.1%), geranyl acetate
(1.37 ± 0.2 to 2.24 ± 0.7%) etc. Among all applied treatments, com-
bined application of MgSO4 @ 1% + ZnSO4 @ 1% recorded highest
citronellol + nerol content (37.79 ± 1.4%) which was 26.4% higher
 600
Table 4
Effect of foliar application of Mg, Cu and Zn sulphate on essential oil composition (%) of damask rose (GC-FID analysis).

Sr. no. Compound RI T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11

Essential oil 0.042 0.053 0.046 0.04 0.041 0.048 0.043 0.044 0.046 0.042 0.045

1 ␣-Pinene 936 0.66 ± 0.5 0.59 ± 0.5 1.04 ± 0.3 1.38 ± 0.0 1.45 ± 0.1 0.79 ± 0.7 0.70 ± 0.8 0.20 ± 0.3 1.23 ± 0.4 0.82 ± 0.1 0.61 ± 0.1

R. Kumar et al. / Industrial Crops and Products 83 (2016) 596–602

2 Sabinene 975 0.22 ± 0.0 0.15 ± 0.1 0.28 ± 0.0 0.37 ± 0.0 0.40 ± 0.0 0.21 ± 0.2 0.08 ± 0.1 0.07 ± 0.1 0.37 ± 0.0 0.08 ± 0.1 0.20 ± 0.0
3 ␤-Pinene 978 0.60 ± 0.3 0.65 ± 0.3 0.87 ± 0.0 1.13 ± 0.1 1.18 ± 0.1 0.76 ± 0.4 0.62 ± 0.3 0.52 ± 0.3 1.06 ± 0.1 0.79 ± 0.1 0.80 ± 0.3
4 Linalool 1101 0.63 ± 0.0 0.85 ± 0.4 1.21 ± 0.0 1.10 ± 0.2 1.35 ± 0.2 0.99 ± 0.4 0.82 ± 0.3 0.69 ± 0.1 1.12 ± 0.0 1.14 ± 0.2 0.85 ± 0.2
5 Z-rose oxide 1111 0.44 ± 0.2 0.63 ± 0.3 1.31 ± 1.1 0.76 ± 0.3 0.87 ± 0.01 0.41 ± 0.4 0.79 ± 1.1 0.17 ± 0.0 0.48 ± 0.4 0.62 ± 0.2 0.42 ± 0.4
6 Phenyl ethyl alcohol 1116 0.57 ± 0.3 0.54 ± 0.2 0.75 ± 0.1 0.69 ± 0.1 1.01 ± 0.3 0.72 ± 0.1 0.59 ± 0.0 0.43 ± 0.0 0.79 ± 0.2 0.75 ± 0.0 0.39 ± 0.1
7 E-rose oxide 1128 0.15 ± 0.0 0.20 ± 0.0 0.22 ± 0.0 0.26 ± 0.0 0.41 ± 0.3 0.21 ± 0.0 0.18 ± 0.0 0.17 ± 0.0 0.23 ± 0.0 0.25 ± 0.0 0.14 ± 0.1
8 Citronellol + nerol 1240 29.89 ± 1.9 34.38 ± 6.7 35.55 ± 4.7 37.39 ± 2.7 35.98 ± 1.5 35.17 ± 1.0 34.39 ± 1.4 34.76 ± 4.5 37.79 ± 1.4 36.09 ± 6.9 33.53 ± 1.2
9 E-geraniol 1264 22.88 ± 0.9 23.44 ± 4.1 27.14 ± 0.0 25.39 ± 3.5 27.82 ± 4.4 25.15 ± 3.1 24.80 ± 4.6 20.87 ± 2.9 25.98 ± 1.0 26.22 ± 2.9 24.45 ± 2.6
10 Citronellol acetate 1352 0.56 ± 0.0 0.51 ± 0.0 0.57 ± 0.0 0.55 ± 0.0 0.61 ± 0.0 0.50 ± 0.0 0.56 ± 0.0 0.59 ± 0.0 0.62 ± 0.1 0.72 ± 0.3 0.62 ± 0.1
11 Eugenol 1356 0.24 ± 0.0 0.22 ± 0.0 0.31 ± 0.0 0.53 ± 0.6 0.10 ± 0.1 0.23 ± 0.0 0.27 ± 0.0 0.27 ± 0.0 0.22 ± 0.0 0.27 ± 0.0 0.36 ± 0.2
12 Geranyl acetate 1380 2.24 ± 0.7 1.80 ± 0.6 2.08 ± 1.0 1.44 ± 0.6 1.75 ± 0.0 1.76 ± 0.5 1.92 ± 0.7 1.90 ± 0.6 1.86 ± 0.2 1.37 ± 0.2 2.20 ± 0.4
13 Methyl eugenol 1402 0.53 ± 0.1 0.47 ± 0.1 0.46 ± 0.0 0.43 ± 0.0 0.43 ± 0.0 0.40 ± 0.0 0.24 ± 0.3 0.52 ± 0.1 0.42 ± 0.0 0.44 ± 0.0 0.50 ± 0.0
14 Caryophyllene 1420 0.47 ± 0.0 0.37 ± 0.1 0.37 ± 0.0 0.34 ± 0.0 0.16 ± 0.2 0.32 ± 0.0 0.34 ± 0.0 0.42 ± 0.0 0.39 ± 0.0 0.34 ± 0.0 0.44 ± 0.0
15 ␣-Guaiene 1436 0.57 ± 0.1 0.50 ± 0.1 0.48 ± 0.0 0.45 ± 0.0 0.42 ± 0.0 0.39 ± 0.0 0.42 ± 0.0 0.53 ± 0.0 0.31 ± 0.3 0.44 ± 0.0 0.96 ± 0.6
16 ␣-Humulene 1457 1.68 ± 0.1 1.64 ± 0.5 1.45 ± 0.3 1.41 ± 0.2 1.30 ± 0.1 1.30 ± 0.0 1.46 ± 0.0 1.86 ± 0.4 1.38 ± 0.0 1.36 ± 0.1 1.28 ± 1.0
17 Pentadecane 1500 0.87 ± 0.1 0.90 ± 0.2 0.73 ± 0.1 0.73 ± 0.1 0.67 ± 0.1 0.69 ± 0.0 0.78 ± 0.0 0.96 ± 0.1 0.73 ± 0.0 0.75 ± 0.0 0.85 ± 0.0
18 Heptadecane 1700 1.87 ± 0.4 1.86 ± 0.6 1.33 ± 0.0 1.48 ± 0.4 1.18 ± 0.6 1.59 ± 0.5 1.72 ± 0.2 1.93 ± 0.2 1.29 ± 0.1 1.52 ± 0.2 1.82 ± 0.5
19 Farnesol 1737 1.00 ± 0.2 1.02 ± 0.5 0.55 ± 0.0 0.77 ± 0.2 0.33 ± 0.5 0.79 ± 0.0 0.94 ± 0.4 0.93 ± 0.1 0.55 ± 0.1 0.39 ± 0.7 0.95 ± 0.6
20 Octadecane 1800 0.29 ± 0.0 0.24 ± 0.0 0.19 ± 0.0 0.20 ± 0.0 0.10 ± 0.1 0.21 ± 0.0 0.21 ± 0.0 0.27 ± 0.0 0.19 ± 0.0 0.20 ± 0.0 0.24 ± 0.0
21 Nonadecene 1873 2.99 ± 0.4 2.57 ± 1.2 1.87 ± 0.4 1.99 ± 0.8 1.73 ± 0.6 2.08 ± 0.5 2.31 ± 0.3 2.89 ± 0.6 1.72 ± 0.1 2.07 ± 0.3 2.53 ± 0.4
22 Nonadecane 1900 16.13 ± 2.6 14.23 ± 5.1 10.37 ± 0.4 10.69 ± 3.7 9.31 ± 4.0 12.91 ± 3.6 13.15 ± 3.8 15.72 ± 2.9 9.04 ± 0.9 11.11 ± 0.9 13.66 ± 2.7
23 Docosane 1999 1.25 ± 0.0 1.19 ± 0.4 0.85 ± 0.0 0.87 ± 0.2 0.86 ± 0.3 0.82 ± 0.7 1.07 ± 0.4 1.32 ± 0.3 0.78 ± 0.0 0.61 ± 0.7 1.17 ± 0.2
24 Heneicosane 2101 6.62 ± 0.9 5.19 ± 2.1 3.75 ± 0.1 3.78 ± 1.2 3.46 ± 1.3 4.58 ± 0.9 4.66 ± 1.8 6.02 ± 1.6 3.36 ± 0.2 3.93 ± 0.0 5.36 ± 1.0
25 Tricosane 2296 1.22 ± 0.9 1.17 ± 0.5 0.74 ± 0.0 0.84 ± 0.2 0.73 ± 0.3 1.01 ± 0.2 0.97 ± 0.5 1.42 ± 0.4 0.82 ± 0.0 0.82 ± 0.1 1.31 ± 0.6
Total 94.32 ± 0.5 95.31 ± 0.3 94.48 ± 0.9 94.98 ± 1.0 93.61 ± 1.6 93.98 ± 2.3 93.98 ± 0.6 95.46 ± 0.3 92.73 ± 0.8 93.11 ± 2.9 95.64 ± 0.8

RI: retention indices; T1Control; T2 MgSO4. (1%); T3:MgSO4 (2%); T4: CuSO4 1%,; T5: CuSO4 (2%); T6: ZnSO4 (1%); T7: ZnSO4 (2%); T8: MgSO4 (1%) + CuSO4 (1%); T9: MgSO4 (1%) + ZnSO4 (1%); T10: CuSO4(1%) + ZnSO4 (1%); T11:
MgSO4 (1%) + CuSO4 (1%) +ZnSO4 (1%); data represent means ± standard deviation of triplicate samples.
 R. Kumar et al. / Industrial Crops and Products 83 (2016) 596–602 601

Table 5
Effect of foliar application of Mg, Cu and Zn sulphate on soil and plant after harvest.

Treatment Soil Plant

Mg (ppm) Cu (ppm) Zn (ppm) Mg (ppm) Cu (ppm) Zn (ppm)

T1: Control 143.4 2.0 6.1 119.24 0.93 4.97

T2: MgSO4 (1%) 142.0 1.5 5.0 130.94 1.41 6.97
T3: MgSO4 (2%) 139.2 1.6 5.5 129.64 1.56 4.87
T4: CuSO4 (1%) 141.6 1.4 4.8 134.25 1.05 5.23
T5: CuSO4 (2%) 123.8 1.9 5.4 132.06 1.33 5.87
T6: ZnSO4 (1%) 142.1 1.7 5.6 126.31 1.56 4.46
T7: ZnSO4 (2%) 142.0 1.8 5.6 126.05 1.39 5.30
T8: MgSO4 (1%) +CuSO4 (1%) 143.1 1.3 4.8 127.28 1.40 4.00
T9: MgSO4 (1%) +ZnSO4 (1%) 143.9 1.7 5.8 122.97 0.96 5.29
T10: CuSO4 (1%) +ZnSO4 (1%) 142.0 2.1 5.3 132.37 1.36 6.32
T11: MgSO4 (1%) +CuSO4 (1%) +ZnSO4 (1%) 139.8 1.8 5.8 136.92 1.13 6.50
SEm (±) 3.6 0.1 0.3 3.32 0.13 0.53

SEm: Standard error of mean.

than control. E-geraniol is the second important constituent of
damask rose essential oil. Application of CuSO4 @ 2% registered
21.6% higher E-geraniol than control. MgSO4 @ 2% (T3) recorded
the highest Z-rose oxide (1.31 ± 1.1%). Phenyl ethyl alcohol is an
important aromatic component of oil, but because of its solubility
in water it is usually lost in the distillation water unless collected
as rose water. Its concentration was higher by the foliar applica-
tion of CuSO4 @ 2%. High concentration of monoterpene alcohols
(citronellol + nerol, E-geraniol, linalool, phenyl ethyl alcohol and
citral), which cause typical rosaceous and freshness character,
and low concentration of hydrocarbons (nonadecane, eicosane and
heneicosane) are the best characteristics of good rose oil (Baser,
1992). Nonadecane, eicosane and heneicosane are not desirable
constituents of rose oil and their percentage was lower with the
combined application of MgSO4 @ 1% + ZnSO4 @ 1% (T9). Different
response of foliar application nutrient has been found in differ-
ent crops. Yadegari and Shakerian (2014) while comparing four
micro nutrients (Cu, Mn, Zn and Fe), found that Fe was more effec-
tive in stimulating the accumulation of caryophyllene, citronellal,
geranial, E-geraniol, geranyl acetate, linalool, neral, thymol and
carvacrol compounds of essential oils of lemon balm (Melissa offic-
inalis). Linalool increased in the herb and seeds of coriander but
dodecenal of herb decreased with treatment of Zn and mixture of
Zn + Fe compared with control (Said-Al Ahl and Omer, 2009). Foliar
application of Cu and Zn is needed more to the developing organs as
they are immobile in plant and cannot be transported to developing
organs, whereas, Mg is moderately mobile. In this case the plants
do not have to take these micronutrients from the soil solution.
The actual amount of these nutrients that reaches the reproductive
tissue can be small and depends on several other factors also.
3.3. Nutrient concentration in soil and plant
Effect of foliar application of nutrients did not affect the con-
centration of Mg, Cu and Zn in soil and plant (Table 5). This
may be due to higher concentration of these nutrients in soil
(Table 2). The highest concentration of Mg was recorded in MgSO4
@ 1% + CuSO4 @ 1% + ZnSO4 @ 1% (136.92 ppm) as compared with
control (119.24 ppm). The availability of Cu was recorded maxi-
mum in MgSO4 @ 1% + CuSO4 @ 1% (1.40 ppm) while minimum was
observed in control (0.93 ppm). The maximum concentration of Zn
was found in MgSO4 @ 1% + CuSO4 @ 1% + ZnSO4 @ 1% (6.50 ppm)
while lowest was found in control. Hasani et al. (2012) reported
similar increase in Zn and Cu concentration in pomegranate leaves
at ZnSO4 @ 0.6% when compared with control conditions. Salih
(2013) also reported similar results in cowpea. In broccoli concen-
tration of Mg, Zn and Cu increased with urea @ 1% (Yildirim et al.,
2007). Zn foliar application significantly increased Zn concentration
in pomegranate leaf (Khorsandi et al., 2009) and Cu concentration in
maize (Aref, 2011). Different foliar application reported significant
effect in concentration of Mg, Cu and Zn in plants.
4. Conclusion
In general, it can be concluded that foliar application of Mg and
Zn sulphate can considerably improve the flower and essential
oil composition of damask rose, particularly if these micronu-
trients were applied together at both stages of maximum leaf
growth and flower bud formation stage under acidic soils of west-
ern Himalayas. Foliar application of ZnSO4 @ 2% recorded 21.9%
and 21.3% higher numbers of flower/plant and flower yield/plant
than control, respectively. Application of MgSO4 @ 1% + ZnSO4 @
1% recorded 26.4% higher citronellol + nerol content (37.79 ± 1.4%)
than control. This indicates that foliar application of Mg and
micronutrients has a significant effect on the composition of the
essential oil. This study provides some useful information about
the effect of application of Mg, Cu, and Zn sulphates on damask
rose production and in that way increases our knowledge about
the effect of foliar application of nutrients on damask rose crop
production under western Himalayas.
Acknowledgements
Authors are grateful to the Director, CSIR-IHBT, Palampur, India
and AcSIR, for providing the necessary facilities during the course
of this study. The authors are also thankful to Mr. Kuldeep Singh
and Mrs. Vijaylata Pathania, technical assistants, for providing nec-
essary help in the field and lab. Help provided by Dr. Neeraj Kumar
in Identification of essential oil compounds is highly appreciated.
Financial assistance from the project AgTech (BSC-110) by CSIR,
New Delhi, India is highly acknowledged. This is IHBT communica-
tion no. 3905.
References
Abd El- Wahab, M.A., 2008. Effect of some trace elements on growth, yield and
chemical constituents of Trachyspermum ammi L. (AJOWAN) plants under Sinai
conditions. Res. J. Agric. Biol. Sci. 4 (6), 717–724.
Adams, P.R., 1995. Identification of Essential Oil Components by Gas
Chromatography/Mass Spectroscopy, 4th ed. Allured Publishing Corporation,
336 Gundersen Drive, Suite A Carol Stream, IL, USA, pp. 60188–62403.
Ahmad, I., Khan, M.A., Qasim, M., Ahmad, R., Randhawa, M.A., 2010. Growth, yield
and quality of Rosa hybrida L. as influenced by various micronutrients. Pak. J.
Agric. Sci. 47 (1), 5–12.
Akhtar, N., Sarker, M.A.M., Akhter, H., Nadda, M.K., 2009. Effect of planting time
and micronutrient as zinc chloride on the growth, yield and oil content of
Mentha piperita. Bangladesh J. Sci. Ind. Res. 44 (1), 125–130.
Anonymous, 2000. Direct Reading on Spectrophotometer HACH DR 2000 (page No.
353 Method No. 8097).
602 R. Kumar et al. / Industrial Crops and Products 83 (2016) 596–602
Aref, F., 2011. Influence of zinc and boron nutrition on copper, manganese and iron
concentrations in maize leaf. Aust. J. Basic Appl. Sci. 5 (7), 52–62.
Bacha, M.A., Sabbah, A.M., Hamady, M.A., 1997. Effect of foliar application of iron,
zinc and manganese on yield, berry quality and leaf mineral composition of
Thompson seedless and roomy red grape cultivars. J. King Saud Univ. Agric. Sci.
9 (1), 127–140.
Baniasad, A., Khajavirad, A., Hosseini, M., Shafei, M.N., Aminzadah, S., Ghavi, M.,
2015. Effect of hydro-alcoholic extract of Rosa damascena on cardiovascular
responses in normotensive rat. Avicenna J. Phytomed. 5 (4), 319–324.
Baser, K.H.C., 1992. Turkish rose oil. Perfum. Flavor. 17, 45–52.
Black, C.A., 1965. Methods of Soil Analysis. Part 2, Chemical and Microbiological
Properties. American Society of Agronomy, Inc. Publisher, Madison, Wisconsin,
USA.
Danyaie, A., Tabaei-Aghdaei, S.R., Jafari, A.A., Matinizadeh, M., Mousavi, A., 2011.
Additive main effect and multiplicative interaction analysis of flower yield in
various Rosa damascena Mill. genotypes across 8 environments in Iran. J. Food
Agric. Environ. 9 (2), 464–468.
Dordas, C., 2009. Foliar application of calcium and magnesium improves growth,
yield, and essential oil yield of oregano (Origanum vulgare ssp. hirtum). Ind.
Crops Prod. 29 (2–3), 599–608.
European Pharmacopoeia, 1975. Maissonneuve Sainte Ruffine, 3, 68.
Grejtovsky, A., Markusova, K., Eliasova, A., 2006. The response of chamomile
(Matricaria chamomilla L.) plants to soil zinc supply. Plant Soil Environ. 52 (1),
1–7.
Hanan Ali, E.A.H., 2007. Effect of Some Fertilization Treatments on Thymus Vulgaris
Plant Cultivated under North Sinai Conditions. Msc. Thesis. Fac, Agric, Zagazig
Univ, Egypt.
Hasani, M., Zamani, Z., Savaghebi, G., Fatahi, R., 2012. Effects of zinc and
manganese as foliar spray on pomegranate yield, fruit quality and leaf
minerals. J. Soil Sci. Plant Nutr. 12 (3), 471–480.
Kazaz, S., Erbas, S., Baydar, H., Dilmacunal, T., Koyuncu, M.A., 2010. Cold storage of
oil rose (Rosa damascena Mill.). flowers. Sci. Hortic. 126 (2), 284–290.
Khorsandi, F., Yazdi, F.A., Vazifehshenas, M.R., 2009. Foliar zinc fertilization
improves marketable fruit yield and quality attributes of pomegranate. Int. J.
Agric. Biol. 11 (6), 766–770.
Kumar, R., Sharma, S., Sood, S., Agnihotri, V.K., 2013a. Agronomic interventions for
the improvement of essential oil content and composition of damask rose
(Rosa damascena Mill.) under western Himalayas. Ind. Crops Prod. 48, 171–177.
Kumar, R., Sharma, S., Sood, S., Agnihotri, V.K., Singh, B., 2013b. Effect of diurnal
variability and storage conditions on essential oil content and quality of
damask rose (Rosa damascena Mill.) flowers in north western Himalayas. Sci.
Hortic. 154, 102–108.
Kumar, R., Sharma, S., Sood, S., Agnihotri, V.K., Singh, V., Singh, B., 2013c.
Evaluation of several Rosa damascena varieties and Rosa bourboniana accession
for essential oil content and composition in western Himalayas. J. Essent. Oil
Res. 26 (3), 147–152.
Marchner, H., 1995. Mineral Nutrition of Higher Plants, 2nd ed. Academic Press,
London.
Mehlich, A., 1984. Mehlich 3 soil test extractant: a modification of the Mehlich 2
extractant. Commun. Soil Sci. Plant Anal. 15 (12), 1409–1416.
Misra, A., Sharma, S., 1991. Critical Zn concentration for essential oil yield and
menthol concentration of Japanese mint. Fertil. Crit. Res. 29 (3), 261–265.
Misra, A., Srivastava, N.K., Kumar, R., Khan, A., 2005. Effect of Palcobutrazol (PP333 )
on flower quality and quantity of Rosa damascena. Commun. Soil Sci. Plant
Anal. 36 (4–6), 477–486.
Misra, A., Dwivedi, S., Srivastava, A.K., Tewari, D.K., Khan, A., Kumar, R., 2006. Low
iron stress nutrition for evaluation of Fe-efficient genotype physiology,
photosynthesis, and essential monoterpene oil(s) yield of Ocimum sanctum.
Photosyntetica 44 (3), 474–477.
Mohamadi, M., Mostafavi, A., Shamspur, T., 2011. Effect of storage on essential oil
content and composition of Rosa damascena Mill. petals under different
conditions. J. Essent. Oil Bear. Plants 14 (4), 430–441.
Nahed Abd El-Aziz, G., Balbaa, L.K., 2007. Influence of tyrosine and zinc on growth,
flowering and chemical constituents of Salvia farinacea plants. J. Appl. Sci. Res.
3 (11), 1479–1489.
Nasiri, Y., Zehtab-Salmasi, S., Nasrullahzadeh, S., Najafi, N., Ghassemi-Golezani, K.,
2010. Effects of foliar application of micronutrients (Fe and Zn) on flower yield
and essential oil of chamomile (Matricaria chamomilla L.). J. Med. Plants Res. 4
(17), 1733–1737.
Parker, D.R., Aguilera, J.J., Thomason, D.N., 1992. Zinc-phosphorus interactions in
two cultivars of tomato (Lycopersicon esculentum L.) grown in
chelator-buffered nutrient solutions. Plant Soil 143 (2), 163–177.
Rao, B.R.R., Rajput, D.K., 2011. Response of palmarosa {Cymbopogon martinii
(Roxb.) Wats. var. motia Burk.} to foliar application of magnesium and
micronutrients. Ind. Crops Prod. 33 (2), 277–281.
Said-Al Ahl, H.A.H., Omer, E.A., 2009. Effect of spraying with zinc and/or iron on
growth and chemical composition of coriander (Coriandrum sativum L.)
harvested at three stages of development. J. Med. Food Plants 1 (2), 30–46.
Said-Al Ahl, H.A.H., Mahmoud, A.A., 2010. Effect of zinc and/or iron foliar
application on growth and essential oil of sweet basil (Ocimum basilicum L.)
under salt stress. Ozean J. Appl. Sci. 3 (1), 97–111.
Salih, H.O., 2013. Effect of foliar fertilization of Fe, B and Zn on nutrient
concentration and seed protein of Cowpea Vigna Unguiculata. IOSR J. Agric. Vet.
Sci. 6 (3), 42–46.
Sawan, Z.M., Hafez, S.A., Basyony, A.E., 2001. Effect of phosphorus fertilization and
foliar application of chelated zinc and calcium on seed, protein and oil yields
and oil properties of cotton. J. Agric. Sci. 136 (2), 191–198.
Shamspur, T., Mohamadi, M., Mostafavi, A., 2012. The effects of onion and salt
treatments on essential oil content and composition of Rosa damascena Mill.
Ind. Crops Prod. 37 (1), 451–456.
Shiriyan, M., Rad, A.H.S., Sayfzadeh, S., Biareh, V., 2014. Influence of Fe and Zn foliar
application on fruit and grain yield of Savory under different plant densities.
Int. J. Plant Environ. Sci. 4 (2), 723–727.
Stein, S.E., 2005. Mass Spectral Database and Software Version 3.02. National
Institute of Standards and Technology (NIST), Gaithersburg, MD, USA.
Subbiah, B.V., Asija, G.L., 1956. A rapid procedure for the determination of available
nitrogen in soils. Curr. Sci. 25 (8), 259–260.
Torun, A., Gültekin, I., Kalayci, M., Yilmaz, A., Eker, S., Cakmak, I., 2001. Effects of
zinc fertilization on grain yield and shoot concentrations of zinc, boron, and
phosphorus of 25 wheat cultivars grown on a zinc deficient and boron-toxic
soil. J. Plant Nutr. 24 (11), 1817–1829.
Van den Berg, A.K., Perkins, T.D., 2004. Evaluation of a portable chlorophyll meter
to estimate chlorophyll and nitrogen contents in sugar maple (Acer saccharum
Marsh.) leaves. For. Ecol. Manag. 200, 113–117.
Volkweiss, S.J., 1991. Fontes e métodos de aplicaçao. In: Ferreira, M.E.; Cruz, M.C.P.
(eds) Micronutrientes na agricultura. Piracicaba: POTAFOS; CNPq,: 391–412.
Yadegari, M., 2012. Chemical composition, antioxidative and antibacterial activity
of the essential oils of wild and cultivated Thymus vulgaris from Iran. Biosci.
Biotechnol. Res. Asia 9 (1), 261–263.
Yadegari, M., Shakerian, A., 2014. Effects of micronutrients foliar application on
essential oils of lemon balm (Melissa officinalis L.). Adv. Environ. Biol. 8 (4),
1063–1068.
Yassa, N., Masoomi, F., Rohani Rankouhi, S.E., Hadjiakhoondi, A., 2009. Chemical
composition and antioxidant activity of the extract and essential oil of Rosa
damascena from Iran, population of Guilan. DARU J. Pharm. Sci. 17 (3), 175–180.
Yildirim, E., Guvenc, I., Turan, M., Karata, A., 2007. Effect of foliar urea application
on quality, growth, mineral uptake and yield of broccoli (Brassica oleracea L.,
var. italica). Plant Soil Environ. 53 (3), 120–128.
Yilmaz, D., Ekinci, K., Dilmacunal, T., Erbas, S., 2011. Effect of harvesting hour on
some physical and mechanical properties of Rosa damascena Mill. J. Sci. Food
Agric. 91 (9), 1585–1590.
Younis, A., Riaz, A., Sajid, M., Mushtaq, N., Ahsan, M., Hameed, M., Tariq, U.,
Nadeem, M., 2013. Foliar application of macro- and micronutrients on the yield
and quality of Rosa hybrida cvs. cardinal and whisky mac. Afr. J. Biotechnol. 12
(7), 702–708.
Youssef, A.A., Ezz El-Din, A.A., Ibrahim, M.E., 2004. Effect of zinc or cadmium on
growth, yield and oil constituents of Ocimum sanctum L. plant under two levels
of fertilizers. Egypt. Pharm. J. 3, 1–17.