Industrial Crops and Products 67 (2015) 257263

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Industrial Crops and Products

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Crop productivity, aroma prole and antioxidant activity in

Pelargonium graveolens LHr. under integrated supply of various
organic and chemical fertilizers
Vineeta Pandey, D.D. Patra
Agronomy and Soil Science Division, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow 226015, India

a r t i c l e

i n f o

Article history:
Received 17 October 2014
Received in revised form 20 January 2015
Accepted 21 January 2015
Available online 5 February 2015
Keywords:
Geranium
Pelargonium graveolens LHr
Crop productivity
Aroma prole
Antioxidant activity

a b s t r a c t
Pelargonium graveolens LHr. is an important commercial crop and source of industrially acclaimed geranium oil. Field experiments were conducted to study the response of integrated nutrient management
on crop productivity, plant biochemical parameters, essential oil yield, oil quality, nutrient content and
antioxidant property in P. graveolens during the years 20122013 and 20132014 in Lucknow, India. Different organic manures and chemical fertilizers in various combinations were applied to the crop. Plant
height increased by 44%, leaf area by 40%, and essential oil yield increased to 106% in poultry manure
(PM) + chemical fertilizer (CF) treatment. The N content was highest in CF followed by PM + CF, which
were about 37% and 34% greater than the control, respectively. Antioxidant activity increased from 1.53%
(control) to 5.94% in PM + CF treatment. Percentage of major aroma compounds increased in the combined application of organic manure and chemical fertilizer. Geraniol (26.08%28.98%) was the major
constituent of essential oil, followed by citronellol (22.86%28.82%), 10-epi--eudesmol (5.15%5.31%),
isomenthone (5.44%6.83%) and linalool (3.01%4.21%).
Study concludes that the combined application of fty percent each of CF (75:30:30 N:P:K kg ha1 )
and PM (2.5 kg ha1 ) gave signicantly higher herb and oil yield with improved aroma prole, enhanced
antioxidant properties and improvement in soil characteristics.
2015 Elsevier B.V. All rights reserved.

1. Introduction
Pelargonium graveolens LHr. popularly known as rose-scented
geranium, belongs to the family Geraniaceae. The crop is cultivated
for its high value essential oil commercially known as geranium
oil. The steam distillation of the fresh above ground parts of the
plant (tender stem, leaves and owers) yields geranium oil with
characteristic rose-like odour. Cosmetics, perfumery and avor
industry, are the primary consumers of geranium oil (Rajeswara
Rao, 2002). The oil possesses antibacterial and antifungal properties as well. The principal constituents of the oil are geraniol
and citronellol, accounting for about 70% of the oil. Other major
chemical components of the geranium oil are linalool, citronellol
formate, geranyl formate, isomenthone and rose oxide. The world
production of the oil is approximately 500750 tonnes per annum
with China being the leading producer followed by Egypt, Algeria,

Corresponding author. Tel.: +91 522 2718536; fax: +91 522 2342666.
E-mail addresses: pandeyvineeta48@gmail.com (V. Pandey),
ddpatra@rediffmail.com (D.D. Patra).
http://dx.doi.org/10.1016/j.indcrop.2015.01.042
0926-6690/ 2015 Elsevier B.V. All rights reserved.

Morocco, Reunion Island and India. India produces around 5 tonnes

of geranium oil against the industry demand of 50 tonnes (Maiti
et al., 2006; Dasuki, 2002). The world trade in geranium oil is
robust and stable (Demarne, 2002). The main importers of geranium oil are France and USA. Globally, the industrial demand of
the oil far exceeds its production levels with FAO estimates of
10% per annum growth in world exports (Anon., 2006). Hence,
there is a commercial opportunity to increase the production of
geranium oil to meet the industrial demand. The cultivable land
availability is always under pressure from food and competitive
commercial crops. Therefore, the challenge is to increase the herb
and oil productivity of such types of crops through improved agricultural practices without increasing their area under cultivation.
The effect of organic and chemical fertilizers in increasing the crop
yield is well documented. However, specic studies on the role
of organic fertilizers and their combinations on P. graveolens are
very few. Singh (2011) reported higher crop and oil yield with a
combination of organic manure (vermicompost) and chemical fertilizer in P. graveolens. Similarly, Ram et al. (2003) observed that the
application of paddy straw mulch gave higher essential oil yield in
P. graveolens. None of the reported studies on P. graveolens have

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V. Pandey, D.D. Patra / Industrial Crops and Products 67 (2015) 257263

experimented with poultry manure (PM) as one of the organic

fertilizers in their studies. Further, the effect of organic manures
and their combinations on the antioxidant properties of P. graveolens have not been evaluated. This study reports the effects of PM
and its combination on herbage yield, oil yield, and oil quality and
antioxidant properties of the P. graveolens.
Practice of using chemical fertilizer alone has its ip side in
terms of soil health and environment (Ahmadian et al., 2011).
Plant nutrition is one of the most important factor affecting quantity and quality of secondary metabolites in plants. Further, the
biological waste management is a serious challenge and its efcient utilization for increasing crop productivity, soil health and
the environment is a viable proposition. In recent decades, growth
of poultry production in developing countries has intensied to
meet the demands of growing population. The poultry waste gives
rise to potential environmental and human health concerns. Globally, poultry manure or litter has been applied to land to enhance
crop production for centuries (Williams et al., 1999). Distillation
and extraction process of medicinal and aromatic plants produces
enormous amounts of distillation waste around the world and
their safe disposal is a major problem. Converting them into highquality vermicompost and using as an organic amendment for
crops is commercially feasible and protable. Recently, medicinal
and aromatic plants have attracted attention for their potential as
antioxidants in reducing free radical induced tissue injury. Therefore, efforts for searching natural antioxidants and increasing their
activity have intensied in recent years.
Keeping the above facts in view, an experiment was conducted
with the objectives to (1) determine the effect of different combinations of organic manure (vermicompost, farmyard manure, poultry
manure) and chemical fertilizers (N, P, K); (2) arrive at the optimum fertilizer regime for higher herbage and essential oil yield
with industrially acceptable quality of geranium oil; (3) study the
effect of various amendments to the plant nutrients content and
its effect on biochemical and antioxidant activity of plant; and (4)
investigate the effect of organic manures and chemical fertilizer on
soil fertility status.
2. Material and methods

10 t ha1 plus N:P:K (75:30:30 kg ha1 ) (6) VC + CF: vermicompost

10 t ha1 plus N:P:K (75:30:30 kg ha1 ) (7) PM + CF: poultry manure
2.5 t ha1 plus N:P:K (75:30:30 kg ha1 ) (8) Control: no fertilizer.
The N, P and K were applied as urea, single superphosphate and
muriate of potash, respectively. Poultry manure, vermicompost,
farmyard manure and chemical fertilizer P and K were applied
to the soil before planting in the plots (3 3 m2 ). Nitrogen at
150 kg N ha1 was added in two split doses: 75 kg N ha1 as a basal
dose and 75 kg N ha1 as a supplementary dose during the peak
growth period. 45 days old seedlings of geranium variety CIMPawan were transplanted in the eld with plant to plant and row
to row spacing of 45 cm.
Plots were irrigated immediately after transplanting for the
proper establishment of the crop in the eld. All intercultural operations were carried out as per need. The crop was harvested after
90 days in mid of April during both the years of the eld experiment. Plant height, leaf area and total plant biomass were recorded
for the plants of each plot.

2.2. Organic matter and soil and plant nutrient analysis

The physicochemical properties of the three different organic
amendments namely, vermicompost, farmyard manure and poultry manure was analyzed and recorded (Table 1). Organic C
was determined by Walkley and Black (1934) method. For the
determination of total N and total P, the organic material was
digested in 1.2:1 H2 SO4 /H2 O2 mixture at 360 C. After digestion,
total N and P were measured by ow injection analyzer. Available P was extracted with sodium bicarbonate and determined
by the molybdo-phosphate blue color method by UVvis spectrophotometer (Olsen et al., 1954). Potassium was extracted with
ammonium acetate and analyzed by ame photometry. Zn, Mn,
Cu, Fe in the soil were extracted with DTPA and analyzed with an
ICPOES. Plant N, P and K was estimated by acid digestion method
and determined by ow injection analyzer.

2.3. Biochemical and antioxidant capacity determination of P.

graveolens leaf extracts

2.1. Experimental detail

A eld experiment was carried out for two consecutive years viz.,
20122013 and 20132014 during DecemberApril at the research
farm of Central Institute of Medicinal and Aromatic Plants, Lucknow, India. The research farm is located at latitude 26.8940 N
and longitude 80.9816 E, with an elevation of about 120 m above
mean sea level. Mean maximum and minimum temperature uctuated from 24.5 to 44.5 C and 6.9 to 27.5 C, respectively. The soil
texture was clay-loam with pH of 8.2 and low in organic carbon
(OC) 0.32%, available nitrogen (N) 135 kg ha1 , available phosphorus 14.2 kg ha1 and available potassium (K2 O) 140 kg ha1 .
The experiment was laid out in a randomized block design with
eight treatments and three replications. The eight treatments were
(1) FYM: farmyard manure at 20 t ha1 (2) VC: vermicompost at
10 t ha1 (3) PM: poultry manure at 5 t ha1 (4) CF: chemical fertilizer (N:P:K at 150:60:60 kg ha1 ) (5) FYM + CF: farmyard manure

Estimation of chlorophyll a and chlorophyll b was done by

Arnon (1949). Carbohydrate content was measured according to
the method of Yemm and Willis (1954) using anthrone reagent.
Ascorbic acid was determined by Keller and Schwager (1977)
method. In brief, 0.5 g of fresh leaf sample was homogenized with
20 mL of extracting solution (oxalic acid + EDTA in distilled water).
It was centrifuged for 15 min at 6000 rpm and the supernatant
collected. On an addition of the supernatant liquid (1 mL) to 2, 6dichlorophenol indophenol (DCPIP), the solution turned pink. The
optical density (OD) of the mixture was taken at 520 nm (Es). After
taking the OD of the mixture, one drop of ascorbic acid was added
to bleach the pink color and again the OD was taken at the same
wavelength (Et). The OD of DCPIP solution was also taken at 520 nm
(Eo). A standard curve was prepared by using different concentration of ascorbic acid following the same method. Concentration of
ascorbic acid was calculated as under:

Table 1
Chemical properties of organic manures.
Parameters

OC (%)

TC (%)

N (%)

Available P mg kg1

Available K mg kg1

Poultry Manure (PM)

Vermicompost (VC)
Farmyard manure (FYM)

30.0 3.10
16.12 1.20
12.7 0.910

33 3.30
19.2 1.30
17.2 0.91

3.2 0.20
1.30 .012
0.82 0.021

57.91 4.72
14.01 1.26
28.01 2.1

192 17.2
99.1 8.50
115 10.3

OC organic carbon; TC total carbon; N nitrogen; P phosphorous; K potassium. All values are the mean of three replications standard error of mean.

V. Pandey, D.D. Patra / Industrial Crops and Products 67 (2015) 257263

Chlorophyll a and b
mg g-1 fresh weight

Chlorophyll a
2
1.8
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0

Table 2
Leaf area, plant height, herbage and oil yield of P. graveolens in different fertilizer
regimes (Average data of 2 years).

Chlorophyll b

e
c
b

a
b

Plant height
(cm)

Treatments Leaf area

(cm2 )

d
d

259

Treatments
Fig 1. Chlorophyll a and chlorophyll b in P. graveolens in different fertilizer regimes
(average data of two years).
Bars are mean of the three replications standard error of mean. Bars followed by
the same letters are not signicantly different at p < 0.05 and different letters show
signicantly different at p < 0.05 based on DMRT test.
FYM farmyard manure; VC vermicompost; PM poultry manure; CF chemical
fertilizer; FYM + CF farmyard manure and chemical fertilizer; PM + CF poultry
manure and chemical fertilizer; VC + CF vermicompost and chemical fertilizer.

Ascorbic acid (mg g1 ) = [Eo (Es Et)] V/W V1 1000,

where W = Weight of the fresh leaf taken, V1 = Volume of the
supernatant taken, V = Total volume of the mixture.
Antioxidant activity (DPPH scavenging activity) was determined according to the method described by Brand-Williams et al.
(1995). The antioxidants react with 1,1-diphenyl-2-picryl-hydrazyl
(DPPH) radical and convert it to 1,1-diphenyl-2-picryl-hydrazine.
The change in color from purple to yellow is used as a measure of
the scavenging potential of antioxidant extracts. Absorbance was
noted at 517 nm by using UVvis spectrophotometer. The capacity
of scavenging free radicals was calculated as follows:
Scavenging activity (%) = {(Control abs sample abs.)/Control
abs.) 100.
2.4. Essential oil extraction
The essential oil extraction was carried out by hydro-distillation
in a Clevenger (1928) type apparatus. Extracted oil was dried with
anhydrous sodium sulphate. The oil yield was calculated in mL of
oil per 100 g of dried material. The collected oil was stored in at
20 C for analysis.
2.5. Gas chromatography (GC) analysis
GC analysis of the essential oils was carried out on a PerkinElmer
Auto System XL gas chromatograph, equipped with DB-5 capillary
column (50 m 0.32 mm i.d., lm thickness 0.25 m) xed in a Varian CP-3800 Gas Chromatograph. The column was programmed
from 60 C, at the rate of 5 C/min, using H2 as carrier gas at constant ow rate of 1 mL/min, a split ratio of 1:40 and S/SL injector and
detector (FID) temperatures were 270 C and 300 C, respectively.
Characterization was achieved on the basis of retention time, elution order, relative retention index using a homologous series of
n-alkanes (C6 C28 ) hydrocarbon, Polyscience Corp., Niles, IL), coinjection with standards in the GC-FID capillary column (Aldrich and
Fluka).
2.6. Statistical analysis
ANOVA analysis was performed for plant characteristics to
detect the signicance of the treatments. Linear correlation coefcient was evaluated among biochemical parameter (chlorophyll

Control
FYM
PM
VC
CF
FYM + CF
VC + CF
PM + CF

42.42
44.41
54.23
46.33
50.04
53.22
61.82
65.35

3.21a
2.27b
3.82c
4.11 d
4.96e
4.29f
5.26 g
4.96 h

45.11
51.7
57.09
53.74
60.11
60.28
65.32
71.11

Herbage yield
(t ha1 )

3.93a
4.65b
3.95c
4.85 d
5.11e
5.32e
5.94f
5.80 g

7.66
8.73
10.24
9.72
11.15
12.22
12.57
16.55

0.61a
0.76b
0.82c
0.88 d
0.11e
0.93f
0.10f
0.13 g

Oil yield
(kg ha1 )
15.23
17.46
20.22
19.48
22.21
30.56
45.16
50.14

1.45a
1.37b
1.96c
1.88 d
1.96e
2.97f
3.97 g
4.55 h

All values are the mean of three replications standard error of mean. Same letters
within the column are not signicantly different at p < 0.05 and different letters are
signicantly different at p < 0.05 based on DMRT test.
FYM farmyard manure; VC vermicompost; PM poultry manure; CF chemical
fertilizer; FYM + CF farmyard manure and chemical fertilizer; VC + CF vermicompost and chemical fertilizer; PM + CF poultry manure and chemical fertilizer.

a and b), vegetative parameter and oil yield. Data was analyzed
by one-way analysis of variance with three replications using the
SPSS statistical software version 20. The signicance of the difference between treatment means was tested using Duncans multiple
range test (DMRT) at p < 0.05 level.
2.7. Principal components analysis (PCA)
PCA was applied for reducing the number of variables (corresponding to plant and soil macro, micronutrients, biomass and oil
yield) of P. graveolens. The aim of the PCA is to produce components
suitable to be used as predictors or response variables in subsequent analysis. The number of factors to keep in data treatment
was evaluated by the scree plot, taking into account the eigenvalues, which should have values greater than one for retaining the
factor in the analysis (Rencher, 1995).
3. Results and discussion
3.1. Effect on vegetative and biochemical parameters (chlorophyll
and carbohydrate content)
Biochemical and vegetative parameters signicantly enhanced
in different fertilization regimes. Contents of chlorophyll a and
chlorophyll b increased considerably in all treatments (Fig 1).
Chlorophyll a content varied from 44% to 63% and chlorophyll b
ranged from 16% to 69%. Highest increase in chlorophyll a and
chlorophyll b was recorded in PM + CF treatment. Biljana and Aca
(2009) reported that photosynthetic potential and plant yield are
directly proportional to each other. Organic manures released N and
Mg, which get incorporated into the porphyrin rings of chlorophyll
molecules resulting in an increase in the content of chlorophyll
a and chlorophyll b in PM + CF treatments (Amujoyegbe et al.,
2007). Higher chlorophyll content results in a rise in the rate of
Table 3
Correlation among chlorophyll (a, b), leaf area, plant height, biomass and oil yield
in P. graveolens.
Chl a
Chl a
Chl b
Leaf area
Plant height
Biomass
Oil yield

Chl b
**

.824
1

Leaf area
**

.749
.865**
1

Chl a chlorophyll a, Chl b chlorophyll b.

**
Correlation is signicant at the p < 0.01.

Plant height
**

.722
.759**
.867**
1

Biomass
**

.746
.895**
.866**
.863**
1

Oil yield
.687**
.901**
.929**
.773**
.939**
1

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V. Pandey, D.D. Patra / Industrial Crops and Products 67 (2015) 257263

25

d
a

15

b
e

20
10

2.7
2.65

2.6
2.55
2.5

2.45

2.4

Carbohydrate
mg g-1 fresh weight

Ascorbic acid
mg g-1 fresh weight

35
30

DPPH acvity

Carbohydrate

DPPH acvty (%)

Ascorbic acid

71
70
69
68
67
66
65
64
63
62

e
c

c
d

b
a

Treatments
Fig 2. Ascorbic acid and carbohydrate content in P. graveolens in different fertilization regimes (average data of two years).
Bars are mean of the three replications standard error of mean. Bars followed by
the same letters are not signicantly different at p < 0.05 and different letters show
signicantly different at p < 0.05 based on DMRT test.
FYM farmyard manure; VC vermicompost; PM poultry manure; CF chemical
fertilizer; FYM + CF farmyard manure and chemical fertilizer; PM + CF poultry
manure and chemical fertilizer; VC + CF vermicompost and chemical fertilizer.

photosynthesis leading to increase in plant height and leaf area by

44% and 40%, respectively in PM + CF treatment (Table 2). A positive and direct association was found among chlorophyll a and
chlorophyll b (r = 0.824**), leaf area and chlorophyll a, chlorophyll
b, (r = 0.749**, r = 0.865**), respectively (Table 3). All biochemical
and vegetative parameters were consistently higher in the organomineral fertilizer treatments as compared to the treatments of
organic or chemical fertilizers alone. These results support the
ndings of Wang and Lin (2002) in strawberry plants. It might
be because the organo-mineral fertilizers provide higher supplies
of both micro and macro nutrients to plants for better crop performance. Carbohydrate content of P. graveolens increased in all
amended plots, being highest in PM + CF treatment (Fig 2) indicating enhancement in the rate of photosynthesis. Carbohydrate
content increased marginally in all the treatments. The results suggest that higher supplies of macro and micronutrients (i.e. higher
levels of N, P, K and Zn, Mg, Mn, Cu) in P. graveolens accelerate the
rate of photosynthesis resulting in higher carbohydrate production
and enhanced plant growth.
3.2. Effect on biomass and essential oil yield
Combined application of organic manures and chemical fertilizer signicantly increased the crop biomass and oil yield. The
biomass yield increased by 37%, 45%, 48% and 73% and essential oil yield by 37%, 66%, 99% and 106% in CF, FYM + CF, VR + CF,
PM + CF treatments, respectively over control. The PM + CF treatment gave highest average total biomass and oil yield per plot
i.e. 16.55 0.13 t ha1 and 50.14 4.55 kg ha1 , respectively, which
were signicantly higher than all other treatments. The control
treatment gave lowest oil yield of 15.23 1.45 kg ha1 (Table 2).
Total biomass of per plot is directly and positively associated with
the total oil yield per plot (r = 0.939**) (Table 3). Our results corroborate the earlier work of Singh et al. (2007) in rosemary and Annrud
et al. (2013) in sorghum and kenaf grass. Increase in biomass and
essential oil yield in combined use of organic and chemical fertilizers may be due to integration of organic manures with chemical
fertilizers that increases the water holding and cation exchange
capacity of the soil. This phenomenon boosts nutrient availability resulting in higher nutrient content leading to higher biomass
and essential oil production. Organic manures alone showed lower
nutrient uptake leading to lower biomass and oil yield. Organic
manures require complex microbial transformation in the soil for
release of the nutrients for the plant. In the case of organic manure
alone, relatively small amounts of nutrient as compared to chemical

Treatments
Fig 3. DPPH scavenging activity of P. graveolens leaf extracts in different fertilizer
regimes (average data of two years).
Bars are mean of the three replications standard error of mean. Bars followed by
the same letters are not signicantly different at p < 0.05 and different letters show
signicantly different at p < 0.05 based on DMRT test.
FYM farmyard manure; VC vermicompost; PM poultry manure; CF chemical
fertilizer; FYM + CF farmyard manure and chemical fertilizer; PM + CF poultry
manure and chemical fertilizer; VC + CF vermicompost and chemical fertilizer.

fertilizer are available to the plant for uptake (Gutser et al., 2005).
Essential oil is a secondary metabolite and according to Croteau
et al. (1972), its production is directly related to the level of photosynthetic activity. Higher biomass due to higher availability of
the nutrients leads to an enhanced level of photosynthetic activity resulting in higher essential oil yield production in the plant.
A comparison of the results of organic manure and chemical fertilizer treatments showed that integrating organic and chemical
fertilizers gave the highest biomass and oil yield. Results showed
that combination of organic and chemical fertilizers are best suited
for higher herb and essential oil yield.
3.3. Essential oil quality
The major chemical components determining the quality
of the P. graveolens oil are linalool citronellol, isomenthone,
geranyl formate, 10-epi--eudesmol and geraniol. Geraniol has
potent antimicrobial activity, and citronellol is the most abundant compound in geranium oil. The analysis of oil obtained from
experimental plants of P. graveolens var. CIM-Pawan demonstrated that the percentage of all the major essential oil components
increased over control in all the fertilizer treatments. Geraniol
content increased from 26% in control to 30% in VC + CF treatment. Citronellol content ranged from 22% to 29% with the highest
percentage in PM + CF followed by FYM + CF (Table 4). Fertilizers
can signicantly modify both the content and composition of the
essential oils. Biofertilization and balanced mineral fertilization
of aromatic plants is an important cultivation factor determining
essential oil quantity and quality. Rate of applied nitrogen modi
es both essential oil content and yield (Nurzynska-Wierdak
et al.,
2013).
Fertilization is the main factor affecting yields of active substances per unit area in herbaceous plants (Kozera et al., 2013).
The quantity and release rate of N, P and K by different treatments affect the quality of the essential oil. According to Koeduka
et al. (2006) nitrogen plays a key role in the biosynthesis of many
organic compounds: amino acids, proteins, enzymes, and nucleic
acids. Amino acids and enzymes play a pivotal role in the biosynthesis of numerous compounds that are essential oil constituents.
Prasad et al. (2012) also observed that phosphorus fertilization signicantly increases oil yield in P. graveolens and the contents of
citronellol and 10-epi--eudesmol. Effect of different combinations

V. Pandey, D.D. Patra / Industrial Crops and Products 67 (2015) 257263

261

Table 4
Principal aroma compounds of P. graveolens under different fertilizer regimes (Average data of 2 years).
Compounds

RIexp

RILit

Linalool
Isomenthone
Citronellol
Geraniol
Geranyl formate
10-epi--Eudesmol

1104
1170
1230
1257
1302
1621

1096
1158
1223
1249
1298
1622

Treatments
Control
3.01
5.44
22.86
26.08
2.71
5.31

FYM
0.22
0.33
1.6
1.8
0.15
0.37

3.63
6.20
25.34
26.01
2.70
5.92

PM

0.21
0.41
1.8
1.7
0.13
0.42

3.85
5.80
24.33
25.8
3.20
5.35

VC

0.24
0.35
1.9
1.60
0.14
0.47

3.50
6.10
26.88
25.66
3.44
6.05

CF

0.25
0.42
1.8
1.4
0.26
0.56

3.65
6.16
26.46
26.25
3.30
5.56

FYM + CF

0.23
0.41
1.6
1.5
0.27
0.48

3.32
6.74
28.67
26.69
2.91
6.15

VC + CF

0.22
0.52
1.8
1.7
0.19
0.52

4.21
6.83
24.59
29.98
3.40
5.91

PM + CF
0.27
0.53
1.7
2.1
0.25
0.46

3.18
6.74
28.82
26.96
3.61
5.80

0.28
0.53
1.9
2.2
0.27
0.49

RIexp - RI on DB-5 capillary columns, RILit - Retention index from Literature (Adams, 2007).
All values are the mean of three replications standard error of mean.
FYM farmyard manure; VC vermicompost; PM poultry manure; CF chemical fertilizer; FYM + CF farmyard manure and chemical fertilizer; VC + CF vermicompost
and chemical fertilizer; PM + CF poultry manure and chemical fertilizer.

of fertilizers on the essential oil quality in various crops is well

documented (Borges et al., 2012; Ganjali et al., 2014; NurzynskaWierdak, 2013; Patruio and Tabara, 2011; Teles et al., 2014). In
sweet basil, content of essential oil in herbs, as well as an increase
in 1,8-cineole in the oil under inuence of the increased rate of
potassium, are reported (Nurzynska-Wierdak et al., 2013).
The quality of the essential oil obtained from the organic plus
chemical fertilizers treatments was superior (as indicated by the
percentage of major chemical components) and commercially
acceptable.
3.4. Effect on antioxidant property
The antioxidant properties are directly related to ascorbic
acid content (vitamin C) and the capacity to scavenge the free
radical 2,2-diphenyl-1-picryl-hydrazyl (DPPH activity). Ascorbic
acid content and antioxidant activity varied with different fertilizer treatments (Fig 2). Organic treatments signicantly increased
ascorbic acid content as compared to CF alone. The increase in
ascorbic acid content may be due to the difference in physicochemical properties and composition of organic and chemical fertilizers
and their effect on soil biota and plant metabolism. The application of CF alone gave higher yields with lower vitamin C content in
tomato (Dumas et al., 2003; Toor et al., 2006). Antioxidant activity
of P. graveolens leaf extract signicantly increased from 1.53% (control) to 5.94% (PM + CF) (Fig 3). Highest value of DPPH was observed
in PM + CF and VC + CF treatment, which was approximately 5.95%
(p < 0.05) higher as compared to the control. In general, the DPPH

values were higher in organic manure plus CF treatments (69%)

with no signicant difference in PM and VC treatments.
3.5. Nutrient content in plant
Content of nutrients in plant was signicantly inuenced by
the different fertilizer regimes (Fig 4). The higher N content was
recorded in CF followed by PM + CF, which were about 37% and
34% higher than the control, respectively. In this experiment, N
released from organic fertilizer alone was lower and inadequate
for economical production of the crop. Similar to N, highest content of K was recorded in CF (40%). K content increased marginally
in all treatments over control. P content increased by 29%24% in CF
and VC + CF treatments, respectively over the control. Results indicate that plants were able to take up most of the N, P and K added
through the use of organic and chemical fertilizers during the two
cultivated seasons. While CF alone may appear an attractive option
for nutrient content in an immediate term, combination of CF and
organic manure is the best strategy for long-term holistic approach
for the cultivation of P. graveolens.
3.6. Effect on soil chemical properties
Different fertilizer regimes affected soil chemical properties signicantly. Data summarized in Table 5 indicate that after harvesting
of P. graveolens, signicantly higher organic C (0.66 .02%) was
accumulated in the soil treated with PM + CF as compared to all
other treatments. Organic C increased with PM + CF by 64% and 56%

Fig 4. Plant nutrient content in P. graveolens in different fertilizer regimes (average data of two years).
Bars are mean of the three replications standard error of mean. Bars followed by the same letters are not signicantly different at p < 0.05 and different letters show
signicantly different at p < 0.05 based on DMRT test.
FYM farmyard manure; VC ermicompost; PM poultry manure; CF chemical fertilizer; FYM + CF farmyard manure and chemical fertilizer; PM + CF poultry manure
and chemical fertilizer; VC + CF vermicompost and chemical fertilizer.

262

V. Pandey, D.D. Patra / Industrial Crops and Products 67 (2015) 257263

Fig 5. Principle component analysis (PCA) for soil chemical properties (C, N, P and K), soil micronutrients (Zn, Fe, Cu and Mn), nutrient content (N, P, K) in plant, plant height,
leaf area, plant biomass and oil yield of P. graveolens.
Chla chlorophyll a; chlb chlorophyll b; PlantN nitrogen content in plant; PlantP phosphorous content in plant; PlantK potassium content in plant; SoilC soil carbon;
SoilN soil nitrogen; SoilP soil phosphorous; SoilK soil potassium; SoilNa soil sodium.

respectively, over control. Available N, P and K in soil after the harvesting were higher in all the treatments as compared to control
during both the years. Highest amount of C and P was added to
soil in case of PM treatment. Thus, application of organic amendments remarkably improved soil fertility status. The improvement
in soil fertility in case of organic manure treatments may be due
to the presence of easily available water-soluble carbon, which
acts as a source of energy for soil micro-organisms (Manna and
Ganguly, 1997). Organic fertilizers enhance plant growth by providing nutrients and improving the effectiveness of the soil by
favorably modifying its water retention and aeration. The results
support the previous ndings of benecial effects of combined
application of organic and inorganic nutrients on soil fertility and
productivity in menthol mint (Patra et al., 2000).

3.7. Principle component analysis

Principal component analysis (PCA) contributes 84% of the total
variance. PC1 contributed 70%, and PC2 contributed 14% of the
total variance. PCA showed that organic C, total N, available P and
available K were the most important factors inuencing the plant
vegetative parameters viz., plant height, leaf area and biomass. PCA
analysis concluded that all soil micro, macronutrients and plant
macronutrients positively affected the total biomass and oil yield
of P. graveolens (Fig 5). In general, the combination of organic and
chemical fertilizers supplied all the required elements for growth
to the plant which enhanced the yield of essential oil by increasing
chlorophyll content, rate of photosynthesis, plant growth, leaf area,
biomass yield, and carbohydrate production.

Table 5
Soil fertility status after harvesting of P. graveolens in different fertilizer regimes (Average data of 2 years).
Macronutrients
Treatments
Control
FYM
PM
VC
CF
FYM + CF
VC + CF
PM + CF

Micronutrients (DTPA extracted)

1

OC (%)

N (kg ha
0.34
0.48
0.52
0.57
0.37
0.52
0.60
0.66

.05a
.05b
.01c
.01c
.05a
.05c
.02 d
.02 d

65.12
69.25
74.25
68.46
75.16
73.23
73.25
78.93

5.5a
6.1b
6.9c
5.7b
6.9 d
7.1c
5.8c
7.2e

K (kg ha
57
63.73
73.0
72.0
63.83
78.13
82.23
93.17

.3.2a
4.5b
5.6c
6.1c
5.9b
6.7 d
7.9e
8.3f

P (kg ha
8.2
11.28
15.20
13.70
10.16
16.23
18.73
24.26

0.72a
.10b
.96c
.87 d
.95e
1.2f
1.3 g
1.2 h

Zn (kg ha1 )
12.78
14.20
15.02
16.08
13.72
16.19
17.36
15.87

.10a
.76b
.93c
.95 d
.89e
.92 d
.92f
.88c

Fe (kg ha1 )
3.03
5.76
6.76
7.2
3.46
8.8
9.24
10.34

.25a
.47b
.50c
.64 d
.28a
.76e
.71f
.83 g

Mn (kg ha1 )
1.93
3.50
3.71
6.06
2.23
6.20
6.76
7.60

.90a
1.2b
.25b
.53c
.18 d
.58c
.62c
.67e

Cu (kg ha1 )
4.10
6.8
7.8
6.02
5.0
13.43
10.33
12.21

.34a
.54b
.65c
.57 d
.46e
1.1f
.98 g
1.1 h

All values are the mean of three replications standard error of mean. Same letters within the column are not signicantly different at p < 0.05 and different letters are
signicantly different at p < 0.05 based on DMRT test.
FYM farmyard manure; VC vermicompost; PM poultry manure; CF chemical fertilizer; FYM + CF farmyard manure and chemical fertilizer; VC + CF vermicompost
and chemical fertilizer; PM + CF poultry manure and chemical fertilizer.

V. Pandey, D.D. Patra / Industrial Crops and Products 67 (2015) 257263

4. Conclusion
The results suggest that combined use of organic manure and
chemical fertilizer gives higher herb yield, higher essential oil yield
with improved quality and increased antioxidant activity with
enhanced carbohydrate and ascorbic acid production in P. graveolens. Combination of poultry manure and chemical fertilizers is
the most suited combination for improving the crop productivity, oil yield, aroma prole, soil health and overall protability and
economics of cultivation of the P. graveolens. Addition of poultry
manure or vermicompost has potential to reduce the use of chemical fertilizers by fty percent with improvement in the herb yield,
soil health and other commercially desirable parameters in the
geranium oil.
Acknowledgments
Authors are thankful to the Director, CSIR-Central Institute of
Medicinal and Aromatic Plants Lucknow, India for providing necessary facilities. Help extended by Dr. C.S Chanotiya and Ms. Anju
Yadav in GC analysis and constructive suggestions by Dr. R.C Padalia
are thankfully acknowledged. to CSIR, New Delhi, India for funding
the research work under their network project Chem-Bio.
References
Adams, R.P., 2007. Identication of essential oil components by gas
chromatography/mass spectrometry, 4th ed. Allured Publishing Corporation,
Carol Stream, Illinois.
Ahmadian, A., Ghanbari, A., Siahsar, B., Haydari, M., Ramroodi, M., Mousavinik,
S.M., 2011. Study of chamomiles yield and its components under drought
stress and organic and inorganic fertilizers using and their residue. J. Microb.
Antimicrob. 3, 2328.
Amujoyegbe, B.J., Opabode, J.T., Olayinka, A., 2007. Effect of organic and inorganic
fertilizer on yield and chlorophyll content of maize (Zea mays L.) and sorghum
Sorghum bicolour (L.) Moench). Afr. J. Biotechnol. 6, 18691873.
Annrud, R., Cihacek, L., Johnson, B.L., Ji, Y., Berti, M.T., 2013. Sorghum and kenaf
biomass yield and quality response to nitrogen fertilization in the Northern
Great Plains of USA. Ind. Crops Prod. 50, 159165.
Anon., 2006. The prole of geranium oil http://www.southinvest.gov.et/
Publications/SSNPR%20draft%20Prole/G/%20Geranium%20Oil. pdf.
Arnon, D.I., 1949. Copper enzymes in isolated chloroplasts: polyphenoxidase in
Beta vulgaris. Plant Physiol. 24, 115.
Biljana, B., Aca, M., 2009. Correlation between nitrogen and chlorophyll content in
wheat (Triticum aestivum L.). Kragujevac J. Sci. 31, 6974.
Borges, L.S., Vieira, M.A.R., Marques, M.O.M., Vianello, F., Lima, G.P.P., 2012.
Inuence of organic and mineral soil fertilization on essential oil of Spilanthes
oleracea cv. Jambuarana. Am. J. Plant Physiol. 7, 135142.
Brand-Williams, W., Cuvelier, M.E., Benset, C., 1995. Use of free radical method to
evaluate antioxidant activity. Lebensm. Wiss. Technol. 28, 2530.
Clevenger, J.H., 1928. Apparatus for the determination of volatile oil. J. Am. Pharm.
Assoc. 17, 346.
Croteau, R., Burbott, A.J., Loomis, W.D., 1972. Biosynthesis of mono- and
sesquiterpenes in peppermint from glucose-14C and 14 CO2 . Phytochemistry
11, 24592467.
Dasuki, U.A., 2002. Pelargonium rosat group. Record from protabase. In: Oyen,
L.P.A., Lemmens, R.H.M.J. (Eds.), Prota (Plant Resources of Tropical
Africa/Ressources Vgtales De Lafrique Tropicale). Wageningen, The
Netherlands.
Demarne, F.-E., 2002. Rose-scented geraniuma pelargonium grown for the
perfume industry. In: Maria Lis-Balchin (Ed.), Geranium and Pelargonium:
History of Nomenclature, Usage and Cultivation. CRC, Press, London, pp.
193207.
Dumas, Y., Dadomo, M., Di Lucca, G., Grolier, P., 2003. Effects of environmental
factors and agricultural techniques on antioxidant content of tomatoes. J. Sci.
Food Agric. 83, 369382.

263

Ganjali, A., Harati, M.P., Kaykhaii, M., Mehdipour, B., Nejad, F.S., Kahkha, M.R.R.,
2014. Investigation of vermicompost fertilizer effect on chemical composition
of essential oil of Ziziphora tenuior in weather conditions of Kahnoojin Iran.
Int. J. Agric. For. 4, 300303.
Gutser, R., Ebertseder, A., Th. Weber, Schrami, M., Schmidhalter, U., 2005.
Short-term and residual availability of nitrogen after long-term application of
organic fertilizers on arable land. J. Plant Nutr. Soil Sci. 168, 439446.
Keller, T., Schwager, H., 1977. Air pollution and ascorbic acid. Eur. J. For. Pathol. 7,
338350.
Koeduka, T., Fridman, E., Gang, D.R., Vassao, D.G., Jackson, B.L., Kosh ChM. Orlova, I.,
Spassova, S.M., Lewis, N.G., Noel, J.P., Baiga, T.J., Dudareva, N., Pichersky, E.,
2006. Eugenol and isoeugenol, characteristic aromatic constituents of spices,
are biosynthesized via reduction of a coniferyl alcohol ester. PNAS 103,
1012810133.
Kozera, W., Majcherczak, E., Barczak, B., 2013. Effect of varied NPK fertilisation on
the yield size, content of essential oil and mineral composition of caraway fruit
(Carum carvi L.). J. Elem. 18, 255267,
http://dx.doi.org/10.5601/jelem.2013.18.2.05.
Maiti, S., Raju, S., Geeta, K.A., Mandal, K.,2006. Good agricultural practices for
patchouli, geranium and lemongrass. National Research Centre for Medicinal
and Aromatic Plants, Anand, Gujarat, India, pp. 815.
Manna, M.C., Ganguly, T.K., 1997. Soil biological activity and yield of crops as
inuenced by compost and inorganic fertilizers under a cereallegume system
in a Typic Haplustert. J. Soil Biol. Ecol. 17, 8894.
Nurzynska-Wierdak, R., 2013. Does mineral fertilization modify essential oil
content and chemical composition in medicinal plants? Acta Sci. Pol. Hortorum
Cultus 12, 316.
Nurzynska-Wierdak, R., Borowski, B., Dzida, K., Zawislak, G., Kowalski, R., 2013.
Essential oil composition of sweet basil cultivars as affected by nitrogen and
potassium fertilization. Turk. J. Agric. For. 37, 427436.
Olsen, S.R., Cole, C.V., Watanabe, F.S., Dean, L.A., 1954. Estimation of Available
Phosphorus in Soils by Extraction with Sodium Bicarbonate. U.S. Department
of Agriculture Circular No. 939, Washington DC.
Patra, D.D., Anwar, M., Chand, S., 2000. Integrated nutrient management and waste
water for restoring soil fertility and productivity of Japanese mint and mustard
sequence in Uttar Pradesh, India. Agric. Ecosyst. Environ. 80, 267275.
Patruio, O., Tabara, V., 2011. Preliminary studies on the volatile oil content of
herbal and the menthol peppermint (Mentha piperita) under the inuence of
fertilization in mineral lovrin. Res. J. Agric. Sci. 43, 139144.
Prasad, A., Kumar, S., Pandey, A., 2012. Microbial and chemical sources of
phosphorus supply modulate the eld and chemical composition of essential
oil of rose-scented geranium (Pelargonium species) in sodic soils. Biol. Fertil.
Soils 48, 117122.
Rajeswara Rao, B.R., 2002. Biomass yield, essential oil yield and essential oil
composition, of rose-scented geranium (Pelargonium species) as inuenced by
row spacing and inter-cropping with corn mint (Mentha arvensis L. f.
piperascens Malinv. Ex Holmes). Ind. Crops Prod. 16, 133144.
Ram, M., Ram, D., Roy, S.K., 2003. Inuence of an organic mulching on fertilizer
nitrogen use efciency and herb and essential oil yields in geranium
(Pelargonium graveolens). Bioresour. Technol. 87, 273278.
Rencher, A.C., 1995. Methods of Multivariate Analysis. John Willey, New York.
Singh, M., 2011. Effect of integrated nutrient management through vermicompost
and inorganic fertilizers on growth yield, nutrient uptake and oil quality of
geranium (Pelargonium graveolens LHer. ex Ait.) grown on alsol. J. Spices
Aromat. Crops 20, 5559.
Singh, M., Ganesha Rao, R.S., Ramesh, S., 2007. Effects of N and K on growth
herbage, oil yield and nutrient uptake pattern of rosemary (Rosmarinus
ofcinalis L.) under semi-arid tropical climate. J. Spices Aromat. Crops 20,
4851.
Teles, S., Pereira, J.A., Muniz de Oliveira, L., Malheiro, R., Machado, S.S., Lucchese,
A.M., Silva, F., 2014. Organic and mineral fertilization inuence on biomass and
essential oil production, composition and antioxidant activity of Lippia
origanoides H.B.K. Ind. Crop. Prod. 59, 169176.
Toor, R.K., Savage, G.P., Heeb, A., 2006. Inuence of different types of fertilizers on
major antioxidant components of tomatoes. J. Food Compos. Anal. 19, 2027.
Walkley, A., Black, I.A., 1934. An Examination of the Degtjareff method for
determining soil organic matter and a proposed modication of the chromic
acid titration method. Soil Sci. 37, 2938.
Wang, S.Y., Lin, H., 2002. Composts as soil supplement enhanced plant growth and
fruit quality of strawberry. J. Plant Nutr. 25, 22432259.
Williams, C.M., Baker, J.C., Sims, J.T., 1999. Management and utilisation of poultry
wastes. Rev. Environ. Contam. Toxicol. 162, 105157.
Yemm, E.W., Willis, A.J., 1954. The estimation of carbohydrate in the plant extract
by anthrone reagent. J. Biochem. 57, 508514.