Article Citation: Azadeh Afrigan, Hossein Azarnivand, Fatemeh Sefidkon, Mohammad Jafari and Mohammad Ali Zare Chahouki Evaluation of environmental factors on essential oil and forage value of Cymbopogon olivieri Journal of Research in Ecology (2017) 5(2): 1095-1112
Evaluation of environmental factors on essential oil and forage value of
Cymbopogon olivieri
Keywords: Cymbopogon olivieri, essential oil, forage value
Authors:
Azadeh Afrigan1,
Hossein Azarnivand2,
Fatemeh Sefidkon3,
Mohammad Jafari2,
Mohammad Ali Zare
Chahouki2
Institution:
1. Department of Reng
Management Science and
Research Branch, Islamic
Azad University, Tehran,
Iran
2. Natural Resources
Faculty, Tehran University,
P.O. Box, Tehran 31585-
4314, Iran
3. Research Institute of
Forests and Rangelands,
Tehran, Iran
Corresponding author:
Hossein Azarnivand
Email ID:
Web Address:
http://ecologyresearch.info/
documents/EC0481.pdf
ABSTRACT: This study was performed to evaluate the environmental factors on essential oil and forage values of Cymbopogon olivieri. In this study, the aerial parts of Cymbopogon olivieri were collected after a time of flowering from 10 natural areas located in the Khuzestan province at two altitudes, and in three replications in the year 2016. Areas included were Chal Gandali, Talkhab e Kalat, Bardmar, Morad Abad, Tembi, Dezful, Indika, Lali, Shoushtar and Izeh. The essential oil compositions were analyzed using gas chromatography-mass spectrometry (GC-MS) analysis. Also, due to phytochemical studies, the plant composition showed Crude Protein (CP), Water Soluble Carbohydrates (WSC), Crude Fiber (CF), Acid detergent fiber (ADF), Neutral Detergent Fiber (NDF) and Total Ash (TA). According to the results, Lali area showed highest mean of essential oil. Analyses of the essential oils showed 21 main identified constituents, including Verbenen, 8/1-Dihydrocodeine , and -2-carene, p-cymene, limonene, p-cymene, Cis-p-mentha-2,8-dien-1-ol, Trans-p-menth, p-mentha-1,5-diene, Methylacetophenone, p-Cymen-8-ol, terpineol, piperitone, germacrene, b-selinene, valencene, f-eti-x-selinene, elemol, Intermedol and y-eudesmot etc. However, Piperitone was present in each area samples more than 50 percentage . The results indicated that essential oils and their chemical compositions of Cymbopogon olivieri are strongly affected by the environmental conditions. Also, according to the results, it was found that essential oil extraction had significant effects on WSC, NDF, CF and total ash. Also there were significant differences between areas.
Dates: Received: 20 Aug 2017 Accepted: 25 Aug 2017 Published: 15 Sep 2017
1095-1112 | JRE | 2017 | Vol 5 | No 2
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An International Scientific Research Journal
Jou
rn
al of R
esearch
in
Ecology
Journal of Research in Ecology
www.ecologyresearch.info
Journal of Research
in Ecology An International
Scientific Research Journal
Original Research
ISSN No: Print: 2319 –1546; Online: 2319– 1554
INTRODICTION
The genus Cymbopogon Spreng as aromatic
grasses is belonging to the Poaceae family (Nair, 1982)
and the genus Cymbopogon comprises two perennial
species in the Flora of Iran, C. Olivieri (Boiss.) Bor and
C. parkeri Stapf., which are distributed in the tropical
regions of Iran including southern parts of Fars, Ker-
man, Hormozgan, Khuzestan, Bushehr and Baluchestan
provinces (Sonboli et al., 2010). C. Olivieri is known as
KahMakki, Potar and Nagerd in different areas which
this plant is gathered. In traditional medicine, leaves and
roots are widely used as antiseptic and for the treatment
of stomachache (Singh et al., 2011). Essential oils of the
Cymbopogon spp. are principally made out of cyclic and
non-cyclic monoterpenes like citral (3,7-dimethyl-2,6-
octadienal; a blend of two isomer geranial and neral),
geraniol, citronellol, citronellal, linalool, elemol, 1,8-
cineole, limonene, b-carophyllene, methylheptenone,
geranyl acetate derivation and geranyl formate (Ito and
Honda, 2007). It is well known that yield and yield
components of plants are determined by a series of fac-
tors including plant genetic (Pirbalouti et al., 2013) cli-
mate, edaphic, elevation, and topography (Loziene and
Venskutonis, 2005) and also an interaction of various
factors (Basu et al., 2009). In an investigation of the
effect of environmental factors on Cymbopogon olivieri
(Boiss.) Bor (Poaceae) in four regions, Mirjalili and
Omidbeigi (2005) concluded that the nearby altitudes of
300-600 m in the regions of Masjid Soleiman and Jiruft
had a greater effect on the function of the essential oil in
the lemongrass (Mirjalili and Omidbeigi, 2005) . Yazda-
ni et al. (2002) reported that the percentage essential oil
of Mentha piperita Stokes (Lamiaceae) varied from
1.45% to 3.2%, and was influenced by different envi-
ronmental factors, such as altitude and the daylight peri-
od (Yazdani et al., 2002). Soil type, climatic regime,
botanical composition and soil improvement practices
as biotic environmental factors have effects on nutri-
tional quality (Corona et al., 1998). The assessment of
feed quality is critical for the predictum of animal per-
formance (Seven and Çerç, 2006). Herbage assessment
infers the depiction of feedstuffs as for their ability to
ensure diverse kinds and levels of production (Juárez et
al., 2004). From a ruminant nutrition point of view, en-
ergy, dry matter digestibility and unrefined protein con-
tents are three of the most imperative segments of herb-
age quality (Malau-Aduli, 2007). It is necessary to ob-
tain information about its chemical composition to im-
prove the quality of the herbage consumed by grazing
animals, which thereafter could be related to its capacity
to satisfy the requirements of the grazing animals.
Mountousis et al. (2011) contemplated altitudinal and
seasonal differences in herbage composition and energy
and protein content of grasslands on Mount Varnoudas,
North-West Greece. They revealed that Crude Protein
(CP) diminished from 106.65 to 72.03 g/kg dry matter
(DM) in the lowlands, from 133.95 to 80.38 g/kg DM in
the center zone and from 127.13 to 74.47 g/kg DM in
sub-alpine grasslands. Metabolizable Energy (ME) con-
tent of the herbage diminished as the growing season
advanced around 19%, 32% and 23% in the lower, mid-
dle and upper altitudinal zone, separately (Mountousis
et al., 2011). According to report of Mountousis et al.,
(2011) ME content of the herbage decreased about 19%,
32% and 23% in the lower, middle and upper altitudinal
zone, respectively (Mountousis et al., 2011). So, the aim
of the present study is to the determine the environmen-
tal effects on the essential oil and forage value of
Cymbopogon olivieri.
MATERIALS AND METHODS
Plant collection and essential oil extraction
In this study, the aerial parts of Cymbopogon
olivieri were collected after a time of flowering from 10
natural areas located in Khuzestan Province at two alti-
tudes, and in three replications at 2016. Territories in-
cluded were ChalGandali, Talkhab e Kalat, Bardmar,
Morad Abad, Tembi, Dezful, Indika, Lali, Shoushtar
Afrigan et al., 2017
1096 Journal of Research in Ecology 2017) 5(2):1095-1112
and Izeh. Basic characterization of region of the present
study shown in the Table 1.
The freshly extracted herb was dried in the la-
boratory setting. A sample (90 g) of the aerial parts of
the plant was extracted using a Clevenger apparatus
through water distillation for three hours. Samples were
dried using anhydrous sodium sulphate (Merck Co. Ger-
many) and then kept in amber vials at 4±1ºC prior to
use. The percentage yield of essential oil is determined
using the formula described by Rao et al. (2005) where
the amount of essential oil recovered (g) was deter-
mined by weighing the oil after moisture was removed
(Rao et al., 2005).
Gas Chromatography–Mass Spectrometry (GC–MS)
analysis
The essential oil analysis was donned using an
analytical gas chromatograph. The GC apparatus was
HP-5MS (5% phenyl methyl silicone and 95% dime-
Afrigan et al., 2017
Journal of Research in Ecology 2017) 5(2):1095-1112 1097
Figure 2. Comparison of means for piperitone percentage between the studied regions
Figure 1. Comparison of the means of essential oil between the regions of the study
Ess
en
tia
l o
il (
g)
thylpolysiloxane), that fitted with DB5 capillary column
(30 m×0.25 mm ID, 0.25 μm film thickness). The col-
umn temperature was initially 60˚C, and then gradually
increased to 120˚C at a 2˚C/min rate, held for three min,
and finally increased to 280˚C and held for 4˚C
(Shahbazi et al., 2015). The carrier gas was helium
Afrigan et al., 2017
1098 Journal of Research in Ecology 2017) 5(2):1095-1112
Table 1. Basic characterization of region of the present study (Meteorological Organization of Khuzestan
Province Data).
Region
Point 1
Latitude
Longitude
Point 2
Latitude
Longitude
Point 3
Latitude
Longitude
Altitudes
from sea
level (m)
Slope
%
Average
Annual
Temperature
(C˚)
Annual
rainfall
(mm)
Annual
evapora-
tion
Chalgandali 321487
3555621
321495
3555627
321485
3555611
328 20 24 350 62
Talkhab-e -
Kalat
323835
3553616
323842
3553623
323830
3553607
301 20 24 340 62
Bardmar 314284
3557151
314293
3557156
314274
3557151
394 55 23 350 63
Morad
Abad
346970
3526940
346980
3526940
346960
3526940
359 5 23 490 62
Tembi 337497
3532764
337505
3532759
337490
3532771
208 45 24 440 64
Dezful 293710
3596903
293712
3596913
293707
3596894
461 5 23 600 58
Indika 354573
3562422
354582
3562427
354565
3562417
724 5 22 600 53
Lali 326321
3578886
326329
3578880
326312
3578891
323 25 23 570 55
Shushtar 319954
3541019
319961
3541012
319946
3541025
148 20 24 290 66
Izeh 377273
3537145
377278
3537136
377268
3537153 768 12 21 630 53
Point 1 Latitude
Longitude
Point 2 Latitude
Longitude
Point 3 Latitude
Longitude
Altitudes
from sea
level (m)
Slope
%
Average
Annual
Temperature
(C˚)
Annual
rainfall
(mm)
Annual
evapora-
tion
Chalgandali 313467
3564142
313475
3564148
313459
3564137
466 5 23 390 61
Talkhab-e-
Kalat
315596
3560341
315601
3560349
315594
3560331
345 5 23 370 62
Bardmar 312168
3557286
312172
3557277
312163
3557295
425 65 23 350 63
Morad
Abad
354143
3562734
354149
3562742
354139
3562725
778 15 22 610 52
Tembi 337813
3522302
337818
3522311
337815
3522292
238 30 25 440 67
Dezful 275695
3614779
275703
3614773
275688
3614786
628 30 23 670 52
Indika 354092
3562779
354100
3562784
354084
3562773
742 15 22 600 52
Lali 326154
3578889
326161
3578882
326148
3578897
393 70 23 580 55
Shushtar 325270
3534168
325277
3534161
325263
3534175
170 40 24 340 66
Izeh 376270
3528603
376276
3528595
376265
3528611
852 20 20 620 54
with 99.99% purity, consistent flow rate of 1ml for each
min, and a split proportion equivalent to 1:20 was uti-
lized. Additionally, the chemical investigation of the
essential oil was done by analytical gas chromatograph
combined with mass spectrometer detector. The capil-
lary column and temperature state of mass spectrometer
detector was comparable with that of gas chromato-
graph that was portrayed previously. The procedure was
worked at 70 eV. The GC-MS investigation was done in
triplicate. Identification of the significant constituents of
the essential oil was done with in view of the correlation
between their Retention Indices (RIs), and that of the
published data. Standard Mass Spectral fragmentation
pattern (Wiley/NBS) and the NIST (National Institute of
Standards and Technology). The percentage of each
essential oil compositions was calculated from the GC
peak areas.
Chemical Analysis
Nitrogen (N) was determined on the basis of
Kjeldahl technique (Bremner and Mulvaney,
1982) ,Crude Protein (CP) was calculated as CP = N%
× 6.25(Jones, 1981). Neutral Detergent Fiber (NDF) and
Acid Detergent Fiber (ADF) were analyzed by the
method of AOAC (2000). Dry Matter Digestibility
(DMD) was estimated using the formula of DMD% =
83.58 – 0.824 ADF% + 2.626N%, suggested by Arzani
et al. (2006). Metabolizable Energy (ME) was calculat-
ed with the equation of ME=0.17 DMD% - 2 reported
by AOAC (2000).
Statistical analysis
The data was statistically analyzed by SPSS
(19.0) software (SPSS, 2010), Means of the traits were
compared by Duncan’s multiple range test at p < 0.05
level.
RESUTS AND DISCUSSION
The means of the essential oil levels in the
Cymbopogon olivieri samples are shown in Figure 1;
according to results, Lali area showed highest mean of
essential oil and significant differences were obtained
between essential oil of Cymbopogon olivieri in Lali
with other area, Also, Tembi didn’t show significant
differences with Shushtar but showed against the three
area viz., Talkhab e Kalat, Morad Abad and Indika
(Figure 1). Higher elevation and colder temperature
provided a better growing condition which led to the
higher accumulation of oil in the leaves. The conse-
quences of different researches showed that altitude is
the most imperative ecological factor affecting oil con-
tent (Kizil, 2010). Our result was obtained at the past
flowering stage but Tajidan et al. (2012) reported essen-
tial oil content decreased with the increase in maturity
stages at harvest, also Ganjewala and Luthra (2007)
studied essential oil biosynthesis of Cymbopogon flexu-
osus (Nees ex Steud) incorporated with (2-14C) acetate.
Their discoveries demonstrated that the yield of essen-
tial oil during the initial leaf growth of 10 to 15 days
was higher (110 to 135 pmol/10 leaves) contrasted with
the yield that was acquired toward the end of leaf
growth cycle of 40 to 50 days (40 to 50 pmol/10 leaves)
(Ganjewala and Luthra, 2007). Research reports demon-
strated that overall essential oil synthesis is related with
the early growth stage in plants, for example, Cymbopo-
gon flexuosus (Singh et al., 1989), Cymbopogon martini
(Sangwan et al., 1982) and Mentha (McCaskill and Cro-
teau, 1995). In general, the yield of essential oil is high-
ly correlated with the yield of biomass.
As indicated by Lewinsohn et al. (1998), the oil
cells of lemongrass were situated inside the paren-
chymatous cells. Different examinations on aromatic
plants showed that essential oils deposited inside the
glandular trichomes (Serrato-Valenti et al., 1997), how-
ever it had been watched that the surface of lemongrass
does not contain glandular trichomes (Lewinsohn et al.,
1998).
Analyses of the essential oils showed 21 main
identified constituents, we can clearly see from table 2
that Verbenen, 8/1-Dihydrocodeine , and -2-carene, p-
Afrigan et al., 2017
Journal of Research in Ecology 2017) 5(2):1095-1112 1099
Afrigan et al., 2017
1100 Journal of Research in Ecology 2017) 5(2):1095-1112
Ta
ble
2.
Id
enti
fica
tio
n o
f es
sen
tia
l o
il c
om
po
siti
on
s o
f C
ymb
op
og
on
oli
vier
iat
the
stu
die
d r
egio
ns
(Alt
itu
des
1)
Ch
al-
G-
An
da
li
Ta
lkh
ab
e K
ala
t
B
ard
ma
r
Mo
rad
Ab
ad
Tem
bi
Ver
ben
en
1.1
8
Ver
ben
en
0.7
8
Ver
ben
en
1.0
2
Ver
ben
en
1.1
3
Ver
ben
en
1.0
3
8/1
-
dih
yd
roco
dei
ne,
0
.27
8/1
-dih
yd
roci
ned
e
0.1
2
8/1
-
dih
yd
roco
dei
ne
0.2
1
&-2
-car
ene
24
.31
8/1
-dih
yd
roci
ned
e
0.1
8
-2-c
aren
e
23
.30
&-2
-car
ene
21
.37
&-2
-car
ene
20
.27
p-c
ym
ene
0.2
5
&-2
-car
ene
22
.36
p-c
ym
ene
0.3
1
p-c
yem
ene
0.2
4
p-c
ym
ene
0.3
4
lim
onene
2.4
2
p-c
ym
ene
0.2
8
lim
onene
2.4
3
lim
onene
2.2
5
lim
onene
2.0
6
p-c
ym
enen
e
0.4
9
lim
onene
2.4
7
p-c
ym
enen
e
0.4
8
p-c
ym
enen
e
0.4
5
p-c
ym
enen
e
0.4
9
Cis
-p-m
enth
-2-e
n
0.1
8
p-c
ym
enen
e
0.5
3
Cis
-p-m
em
th-
2/8
0
.25
Cis
-p-m
enth
-2-e
n
0.1
2
Cis
-p-m
enth
-2-
en
1.3
3
Cis
-p-m
em
th-2
/8
0.2
8
Cis
-p-m
enth
-2-e
n
0.1
8
Tra
ms-
p-m
enth
0
.60
Cis
-p-m
em
th-2
/8
0.2
9
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-p-m
em
th-
2/8
0
.29
Tra
ms-
p-m
enth
0
.63
Cis
-p-m
em
th-2
/8
0.3
9
p-m
enth
a-5
/1-
die
m
0.5
9
Tra
ms-
p-m
enth
0
.45
Tra
ms-
p-m
enth
0
.59
p-m
enth
a-5
/1-
die
m
0.5
1
Tra
ms-
p-m
enth
0
.48
Met
hyl-
acet
op
hen
one
0.8
4
p-m
enth
a-5
/1-d
iem
0
.46
p-m
enth
a-5
/1-
die
m
0.7
5
Met
hyl-
acet
op
hen
s 0
.85
p-m
enth
a-5
/1-
die
m
0.5
7
pip
erit
one
58
.88
Met
hyl-
acet
op
hen
one
0.9
0
Met
hyl-
acet
op
hen
s 0
.82
Cym
en
-8-0
1
0.2
2
Met
hyl-
acet
op
hen
s 0
.89
b-s
elin
ene
0.1
5
Cym
en
-8-0
1
0.1
9
Cym
en
-8-0
1
0.2
5
terp
ineo
l 0
.26
Cym
en
-8-0
1
0.2
1
val
ence
ne
0.4
1
terp
ineo
l 0
.24
terp
ineo
l 0
.26
pip
erit
one
58
.20
terp
ineo
l 0
.28
f-et
i-x-s
elin
ene
0.1
8
pip
erit
one
57
.59
pip
erit
one
59
.25
b-s
elin
ene
0.1
4
pip
erit
one
59
.73
elem
ol
1.0
0
yer
mac
rene
0.1
8
val
ence
ne
0.2
6
val
ence
ne
0.4
0
b-s
elin
ene
0.1
4
y-e
ud
esm
ol
0.6
4
b-s
elin
ene
0.3
4
elem
ol
1.3
8
f-et
i-x-s
elin
ene
0.1
9
val
ence
ne
0.3
7
Inte
rmed
ol
7.5
1
val
ence
ne
0.4
7
y-e
ud
esm
ot
0.8
9
elem
ol
0.9
9
f-et
i-x-s
elin
ene
0.1
9
f-et
i-x-s
elin
ene
0.2
0
Inte
rmed
ol
8.9
9
y-e
ud
esm
ot
0.5
6
elem
ol
0.9
9
elem
ol
1.6
6
y-e
ud
esm
ot
0.5
9
y-e
ud
esm
ot
1.1
9
Inte
rmed
ol
7.1
1
Inte
rmed
ol
9.3
6
Su
m
99
.01
9
8.8
4
9
9.4
5
9
2.0
1
9
8.9
6
Co
nti
nue
....
..
Afrigan et al., 2017
Journal of Research in Ecology 2017) 5(2):1095-1112 1101
Co
nti
nue
...
D
ezfu
l
In
dik
a
L
ali
Sh
ush
tar
Izeh
Ver
ben
en
1.2
7
Ver
ben
en
1.0
1
Ver
ben
en
0.6
9
Ver
ben
en
0.9
2
Ver
ben
en
1.0
4
8/1
-dih
yd
roco
dei
ne
0.2
6
8/1
-dih
yd
roco
dei
ne
0.1
2
8/1
-dih
yd
roco
dei
ne
0.2
0
8/1
-dih
yd
roco
dei
ne
0.1
7
8/1
-dih
yd
roco
dei
ne
0.1
4
&-2
-car
ene
23
.87
&-2
-car
ene
20
.04
&-2
-car
ene
19
.92
&-2
-car
ene
18
.01
&-2
-car
ene
18
.52
p-c
ym
ene
0.3
7
p-c
ym
ene
0.2
5
p-c
ym
ene
0.3
0
p-c
ym
ene
0.2
8
p-c
ym
ene
0.3
1
lim
onene
2.8
3
lim
onene
2.0
4
lim
onene
2.3
2
lim
onene
1.9
7
lim
onene
2.2
1
p-c
ym
enen
e
0.4
9
p-c
ym
enen
e
0.5
5
p-c
ym
enen
e
0.5
7
p-c
ym
enen
e
0.4
8
p-c
ym
enen
e
0.5
5
Cis
-p-m
em
th-2
/8
0.2
6
Cis
-p-m
enth
-2-e
n
0.1
3
Cis
-p-m
enth
-2-e
n
0.2
1
Cis
-p-m
enth
-2-e
n
0.2
4
Cis
-p-m
enth
-2-e
n
0.2
2
Tra
ms-
p-m
enth
0
.56
Cis
-p-m
em
th-2
/8
0.3
2
Cis
-p-m
em
th-2
/8
0.3
4
Cis
-p-m
em
th-2
/8
0.3
4
Cis
-p-m
em
th-2
/8
0.3
6
p-m
enth
a-5
/1-d
iem
0
.62
Tra
ms-
p-m
enth
0
.59
Tra
ms-
p-m
enth
0
.43
Tra
ms-
p-m
enth
0
.57
Tra
ms-
p-m
enth
0
.41
Met
hyl-
acet
op
hen
s 0
.88
p-m
enth
a-5
/1-d
iem
0
.56
p-m
enth
a-5
/1-d
iem
0
.64
p-m
enth
a-5
/1-d
iem
0
.63
p-m
enth
a-5
/1-d
iem
0
.53
Cym
en
-8-0
1
0.2
2
Met
hyl-
acet
op
hen
s 0
.88
Met
hyl-
acet
op
hen
s 1
.01
Met
hyl-
acet
op
hen
s 0
.95
Met
hyl-
acet
op
hen
s 0
.97
pip
erit
one
58
.22
Cym
en
-8-0
1
0.2
0
Cym
en
-8-0
1
0.2
4
Cym
en
-8-0
1
0.2
9
Cym
en
-8-0
1
0.2
8
b-s
elin
ene
0.1
5
terp
ineo
l 0
.29
terp
ineo
l 0
.26
terp
ineo
l 0
.27
terp
ineo
l 0
.82
val
ence
ne
0.2
7
pip
erit
one
62
.64
pip
erit
one
66
.61
pip
erit
one
64
.62
pip
erit
one
66
.53
f-et
i-x-s
elin
ene
0.2
0
val
ence
ne
0.8
8
y-e
ud
esm
ot
0.3
7
b-s
elin
ene
0.1
2
val
ence
ne
0.2
2
elem
ol
0.8
1
elem
ol
0.7
0
Inte
rmed
ol
3.7
5
val
ence
ne
0.3
3
elem
ol
0.4
0
y-e
ud
esm
ot
0.5
4
Inte
rmed
ol
7.4
9
f-et
i-x-s
elin
ene
0.1
7
y-e
ud
esm
ot
0.3
2
Inte
rmed
ol
7.2
0
elem
ol
0.7
3
Inte
rmed
ol
5.6
0
y-e
ud
esm
ot
0.5
5
Inte
rmed
ol
7.1
9
sum
9
9.0
1
9
8.7
0
9
7.8
6
9
8.8
3
9
9.4
4
cymene, limonene, p-cymene, Cis-p-mentha-2,8-dien-1-
ol, Trans-p-menth, p-mentha-1,5-diene, Methylaceto-
phens, p-cymen-8-ol, terpineol, piperitone, germacrene,
b-selinene, valencene, f-eti-x-selinene, elemol, Inter-
medol and y-eudesmot were obtained. A study on the
quality and quantity of essential oil showed
Afrigan et al., 2017
1102 Journal of Research in Ecology 2017) 5(2):1095-1112
Ta
ble
3.
Iden
tifi
cati
on
of
esse
nti
al
oil
co
mp
osi
tio
ns
of
Cym
bo
pog
on
oli
vier
i a
t st
ud
ied
reg
ion
s (A
ltit
ud
es 2
)
C
ha
lGa
nd
ali
Ta
lkh
-ab
- e
Ka
lat
B
ard
ma
r
Mo
rad
Ab
ad
Tem
bi
Ver
ben
en
1.0
0
Ver
ben
en
0.7
6
Ver
ben
en
1.0
1
Ver
ben
en
1.1
1
8/1
-dih
yd
roco
dei
ne
0.1
9
8/1
-dih
yd
roco
dei
ne
0.2
0
8/1
-dih
yd
roco
dei
ne
0.1
8
8/1
-dih
yd
roco
dei
ne
0.2
7
&-2
-car
ene
16
.30
&-2
-car
ene
19
.59
&-2
-car
ene
17
.84
&-2
-car
ene
29
.26
p-c
ym
ene
0.2
9
p-c
ym
ene
0.1
5
p-c
ym
ene
0.2
1
lim
onene
2.4
8
lim
onene
1.8
2
lim
onene
1.7
9
lim
onene
1.9
0
p-c
ym
enen
e
0.5
8
p-c
ym
enen
e
0.5
1
p-c
ym
enen
e
0.5
9
p-c
ym
enen
e
0.5
3
Cis
-p-m
em
th-2
/8
0.3
5
Cis
-p-m
enth
-2-e
n
0.2
2
Cis
-p-m
enth
-2-e
n
0.1
5
Cis
-p-m
enth
-2-e
n
0.1
5
Tra
ms-
p-m
enth
0
.63
Cis
-p-m
em
th-2
/8
0.3
4
Cis
-p-m
em
th-2
/8
0.3
7
Cis
-p-m
em
th-2
/8
0.3
3
p-m
enth
a-5
/1-d
iem
0
.47
Tra
ms-
p-m
enth
0
.66
Tra
ms-
p-m
enth
0
.60
Tra
ms-
p-m
enth
0
.71
Met
hyl-
acet
op
hen
s 0
.92
p-m
enth
a-5
/1-d
iem
0
.77
p-m
enth
a-5
/1-d
iem
0
.49
p-m
enth
a-5
/1-d
iem
0
.68
Cym
en
-8-0
1
0.2
0
Met
hyl-
acet
op
hen
s 0
.87
Met
hyl-
acet
op
hen
s 1
.00
Met
hyl-
acet
op
hen
s 0
.96
terp
ineo
l 0
.27
Cym
en
-8-0
1
0.3
0
Cym
en
-8-0
1
0.2
6
Cym
en
-8-0
1
0.2
7
pip
erit
one
56
.25
terp
ineo
l 0
.29
terp
ineo
l 0
.29
terp
ineo
l 0
.70
val
ence
ne
0.2
3
pip
erit
one
64
.95
pip
erit
one
63
.32
pip
erit
one
60
.28
f-et
i-x-s
elin
ene
0.1
5
b-s
elin
ene
0.1
4
b-s
elin
ene
0.1
1
val
ence
ne
0.3
0
elem
ol
0.6
7
val
ence
ne
0.4
2
val
ence
ne
0.3
1
elem
ol
1.2
9
y-e
ud
esm
ot
0.3
9
f-et
i-x-s
elin
ene
0.2
0
f-et
i-x-s
elin
ene
0.1
6
y-e
ud
esm
ot
0.7
6
Inte
rmed
ol
5.0
4
elem
ol
0.7
0
elem
ol
0.8
9
Inte
rmed
ol
11
.17
y-e
ud
esm
ot
0.5
3
y-e
ud
esm
ot
0.5
9
Inte
rmed
ol
7.8
0
Inte
rmed
ol
7.3
9
Su
m
98
.29
9
9.0
0
9
9.2
7
9
9.2
8
Co
nti
nue.
..
Afrigan et al., 2017
Journal of Research in Ecology 2017) 5(2):1095-1112 1103
Co
nti
nue
…
D
ezfu
l
In
dik
a
L
ali
Sh
ush
tar
Iz
eh
Ver
ben
en
1.0
3
Ver
ben
en
0.8
1
Ver
ben
en
0.1
9
Ver
ben
en
0.9
2
Ver
ben
en
1.0
3
8/1
-dih
yd
roco
dei
ne
0.2
1
8/1
-
dih
yd
roco
dei
ne
0.1
3
8/1
-dih
yd
roco
dei
ne
18
.17
8/1
-dih
yd
roco
dei
ne
0.1
7
8/1
-dih
yd
roco
dei
ne
0.1
0
&-2
-car
ene
19
.97
&-2
-car
ene
14
.59
&-2
-car
ene
0.3
2
&-2
-car
ene
18
.01
&
-2-c
aren
e 1
8.9
7
p-c
ym
ene
0.3
5
p-c
ym
ene
0.2
0
p-c
ym
ene
1.9
7
p-c
ym
ene
0.2
8
p-c
ym
ene
0.3
2
lim
onene
2.2
9
lim
onene
1.6
3
lim
onene
0.2
8
lim
onene
1.9
7
lim
onene
2.1
7
p-c
ym
enen
e
0.5
1
p-c
ym
enen
e
0.5
4
p-c
ym
enen
e
0.1
6
p-c
ym
enen
e
0.4
8
p-c
ym
enen
e
0.5
7
Cis
-p-m
enth
-2-e
n
0.2
2
Cis
-p-m
enth
-2-
en
0.1
5
Cis
-p-m
enth
-2-e
n
0.1
9
Cis
-p-m
enth
-2-e
n
0.2
4
Cis
-p-m
enth
-2-e
n
0.1
6
Cis
-p-m
em
th-2
/8
0.3
2
Cis
-p-m
em
th-
2/8
0
.34
Cis
-p-m
em
th-2
/8
0.3
1
Cis
-p-m
em
th-2
/8
0.3
4
Cis
-p-m
em
th-2
/8
0.3
8
Tra
ms-
p-m
enth
0
.43
Tra
ms-
p-m
enth
0
.53
Tra
ms-
p-m
enth
1
.17
Tra
ms-
p-m
enth
0
.57
T
ram
s-p-m
enth
0
.63
p-m
enth
a-5
/1-d
iem
0
.74
p-m
enth
a-5
/1-
die
m
0.5
4
p-m
enth
a-5
/1-d
iem
0
.60
p-m
enth
a-5
/1-d
iem
0
.63
p
-menth
a-5
/1-d
iem
0
.55
Met
hyl-
acet
op
hen
s 0
.92
Met
hyl-
acet
op
hen
s 1
.00
Met
hyl-
acet
op
hen
s 6
.13
Met
hyl-
acet
op
hen
s 0
.95
M
eth
yl-
acet
op
hen
s 1
.02
Cym
en
-8-0
1
0.2
7
Cym
en
-8-0
1
0.2
5
pip
erit
one
61
.70
Cym
en
-8-0
1
0.2
9
Cym
en
-8-0
1
0.2
9
terp
ineo
l 0
.27
terp
ineo
l 0
.30
b-s
elin
ene
0.2
9
terp
ineo
l 0
.27
te
rpin
eol
0.2
8
pip
erit
one
62
.66
pip
erit
one
65
.92
val
ence
ne
0.2
4
pip
erit
one
64
.62
p
iper
ito
ne
65
.66
val
ence
ne
0.2
5
yer
mac
rene
0.3
7
y-e
ud
esm
ot
0.4
8
b-s
elin
ene
0.1
2
val
ence
ne
0.2
0
f-et
i-x-s
elin
ene
0.1
4
b-s
elin
ene
0.3
6
Inte
rmed
ol
4.8
1
val
ence
ne
0.3
3
elem
ol
0.5
8
elem
ol
0.8
0
val
ence
ne
0.4
8
f-et
i-x-s
elin
ene
0.1
7
y-e
ud
esm
ot
0.4
6
y-e
ud
esm
ot
0.6
2
f-et
i-x-s
elin
ene
0.2
1
elem
ol
0.7
3
Inte
rmed
ol
5.6
7
Inte
rmed
ol
6.9
1
elem
ol
1.3
5
y-e
ud
esm
ot
0.5
5
y-e
ud
esm
ot
0.9
4
Inte
rmed
ol
7.1
9
Inte
rmed
ol
8.3
8
Su
m
98
.91
9
9.0
2
9
7.0
2
9
8.8
3
9
9.0
6
that similar observation was reported in the production
of essential oil of lemongrass in relation to the leaf age
(Singh et al., 1989).
At level 1, The percentages of identified compo-
nents in the essential of the plant at the ChalGandali,
Talkhab e Kalat, Bardmar, Morad Abad, Tembi, Dezful,
Indika, Lali, Shoushtar and Izeh were 99.01, 98.84,
99.45, 92.01, 98.96, 99.01, 98.7, 97.86, 98.83 and
99.44%, respectively Table 2.
Also at level 2, identified components showed
98.29, 99.0, 99.27, 99.28, 98.91, 99.02, 97.02, 98.83
and 99.06 for ChalGandali, Talkhab e Kalat, Bardmar,
Morad Abad, Tembi, Dezful, Indika, Lali, Shoushtar
and Izeh, respectively Table 3. According to Raina et al.
(2003) on the essential oil composition of Cymbopogon
martinii from the different places of India, three sam-
ples of geraniol (67.6–83.6%) was the major constituent
and, although the composition of the three oils were
similar, quantitative differences were observed in the
concentration of some constituents (Raina et al., 2003).
Gas Chromatograph (Thermo-UFM) for the regions
showen at figure 3 to 12.
In this study, piperitone as main composition
varied between 57.99 and 66.10%, the mean comparison
conducted through the Duncan test and results showed
significant differences between regions for piperitone
percentage Figure 2. Highest mean was observed by
Izeh samples and lowest means were obtained by Tembi
region. In this order, Tajidan et al. (2012) reported that
among 13 compounds, only 7 compounds (β-myrcene, 3
-undecyne, neral, geranial, nerol, geranyl acetate and
juniper camphor) had concentration greater than 1%. As
indicated by Schaneberg and Khan (2002), the essential
oil of lemongrass contains for the most part geranial and
neral. Other compounds isolated, for example, β-
myrcene, ocimene, β-ocimene, linalool, citronellal, cit-
ronellol, caryophyllene and β-pinene, were available as
minor segments (Torres and Ragadio, 1996).
Afrigan et al., 2017
1104 Journal of Research in Ecology 2017) 5(2):1095-1112
Table 4. Means comparison between areas and essential oil extraction stages
Areas
Essential oil
extraction CP WSC DMD ADF NDF CF ASH
Chal-
Gandali
Before 5.34 a 10.47 d 46.13 i 45.11 cd 65.01 h 60.89 f 3.93 cde
After 4.78 c 4.08 g 46.69 hi 45.27 cd 75.27 e 70.76 ab 3.25 efg
TalkhabK
alat
Before 3.98 e 8.10 e 50.32 a 41.40 gh 70.00 g 61.10 f 4.08 b-e
After 5.14 b 11.37 cd 48.22 e 40.80 h 60.12 i 56.25 h 5.36 a
Before 4.90 bc 11.36 cd 47.71 ef 42.61 fg 64.40 h 60.28 f 3.88 de
Bardmar After 4.37 d 6.59 f 49.32 cd 43.15 ef 77.05 d 68.83 c 3.56 efg
Morad
Abad
Before 3.31 g 10.57 d 47.13 gh 42.80 fg 57.87 j 56.57 h 5.26 a
After 3.31 g 4.80 g 46.71 h 46.15 bc 81.00 a 67.64 d 4.62 a-d
Tembi
Before 2.75 i 10.72 d 48.96 cd 40.41 h 57.38 j 54.03 j 4.93 abc
After 1.43 k 6.40 f 43.63 k 50.00 a 81.70 a 70.60 ab 2.88 fg
Dezful
Before 3.53 f 12.49 bc 49.91 ab 40.22 h 55.69 k 55.37 i 4.48 a-e
After 3.60 f 4.70 g 46.46 hi 46.21 bc 80.29 b 71.27 a 2.95 efg
Indika
Before 1.32 k 15.01 a 48.77 de 37.66 i 55.40 k 53.80 k 3.60 ef
After 1.21 k 8.05 e 46.83 gh 42.98 fg 71.58 f 66.06 e 2.68 g
Lali
Before 4.21 de 4.49 g 44.52 j 47.68 b 78.04 c 71.12 a 3.19 efg
After 5.18 ab 10.80 d 47.27 fg 43.35 ef 60.79 i 57.16 g 4.84 abc
Shushtar
Before 2.27 j 13.53 b 47.66 f 40.52 h 51.20 l 54.18 j 4.13 b-e
After 4.82 c 5.33 fg 48.92 de 44.68 de 78.09 c 69.15 b 3.97 cde
Izeh
Before 3.79 ef 11.99 bcd 49.44 bc 42.13 fg 59.83 i 57.93 g 4.54 a-d
After 3.00 h 6.26 f 46.21 i 46.71 bc 75.25 e 68.56 c 3.30 efg
Crude Protein (CP)
Results showed that that there was no signifi-
cant difference between before and after essential oil
extraction, but studied areas showed significant differ-
ences at 5% statistical level and the highest CP content
(5.06%) was in Chalgandali samples, also Indika
showed lowest content (1.26%). Arzani et al. (2001)
revealed that locations had tremendous impart on forage
quality due to the distinctions in soil and climate attrib-
utes. The CP content declined through the growing sea-
son, as a response to tissue ageing, Corona et al. (1998)
reported that crude protein declines with the stage of
maturation. Consequently, the reduction of crude pro-
tein content, which was observed in the lower zone,
could be attributed to the different phonological stages
of vegetation. It has been recommended (Bxuton, 1996)
that the aging of plants deduces in the decrease of crude
protein content of leaves and stems, however it addi-
tionally prompts a more noteworthy extent of stems,
which contains less crude protein than leaves.
Water Soluble Carbohydrates (WSC)
WSC value differed among two stages of before
Afrigan et al., 2017
Journal of Research in Ecology 2017) 5(2):1095-1112 1105
Figure 3. Gas Chromatograph (Thermo-UFM) for the Indika sample
Figure 4. Gas Chromatograph (Thermo-UFM) for the Dezful sample
and after essential oil extraction (P0.05), but Significant differences
were obtained between ADF of areas, Lali and Chal-
gandali with 45.5% and 45.19 showing highest value of
ADF and lowest mean was observed by Indika area
(40.32%). There were significant differences in NDF
content of between before and after essential oil extrac
Afrigan et al., 2017
1106 Journal of Research in Ecology 2017) 5(2):1095-1112
Figure 5. Gas Chromatograph (Thermo-UFM) for the Izehsample
Figure 6. Gas Chromatograph (Thermo-UFM) for the Lali sample
tion (P
Afrigan et al., 2017
1108 Journal of Research in Ecology 2017) 5(2):1095-1112
they showed highest (65.82%) and lowest (58.6) means,
respectively. In this order, Marshal et al. (2005) showed
that CP was positively associated with forage growth (P
Afrigan et al., 2017
Journal of Research in Ecology 2017) 5(2):1095-1112 1109
differences when lemongrass was harvested from differ-
ent areas. The highest percentage of essential oil was
obtained when lemongrass was collected from Lali area.
There were 21 chemical compounds detected in the es-
sential oil of lemongrass. However, piperitone of the
compounds was present in each area samples at more
than 50 percentage. According to the results, it was
founded that essential oil had significant effects on
WSC, NDF,CF and total ash. Also there were signifi-
cant differences between areas. Altitude has effects on
maturity time, so, chemical component change accord-
ing to growth stages.
Figure 11. Gas Chromatograph (Thermo-UFM) for the Tembi sample
Figure 12. Gas Chromatograph (Thermo-UFM) for the Shushtar sample
Afrigan et al., 2017
1110 Journal of Research in Ecology 2017) 5(2):1095-1112
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