Bioresource Technology 92 (2004) 311–319
Effect of different organic manures/composts on the herbageand essential oil yield of Cymbopogon winterianus and theirinfluence on the native AM population in a marginal alfisol
Tanu a, Anil Prakash b, Alok Adholeya a,*
a Centre For Mycorrhizal Research, The Energy Research Institute, Habitat Place, Lodhi Road, New Delhi 110003, Indiab Department of Biotechnology, Barkatullah University, Bhopal, Madhya Pradesh 462026, India
Received 8 March 2003; received in revised form 20 May 2003; accepted 27 June 2003
Abstract
Four organic amendments: leaf compost (LC), vegetable compost (VC), poultry manure (PM) and sewage sludge (SSL) applied
at four doses (40, 80, 100 and 120 t ha�1) were evaluated for their effect on the herbage yield, essential oil content and inoculum
potential (IP) of native arbuscular mycorrhizal fungi (AMF) on three varieties of Java citronella, Cymbopogon winterianus Jowitt
(Manjusha, Mandakini, and Bio-13). PM applied at 100 t ha�1 followed by SSL increased the herbage, essential oil content and dry
matter yield significantly. Bio-13 performed better and produced the highest herbage, essential oil and dry matter yield. The type and
dose of the various organic amendments also significantly influenced the indigenous AMF infectious propagules in soil. Highest
number of AMF propagules were recorded in the LC amended plots in all the three varieties. Amongst the varieties, highest native
mycorrhizal inoculum was recorded in the Bio-13. Least number of AM infectious propagules were recorded in the Mandakini
plants grown in 40 t ha�1 SSL.
� 2003 Elsevier Ltd. All rights reserved.
Keywords: Compost/manures; Cymbopogon sp.; Arbuscular mycorrhizal fungi; Inoculum potential; Herbage yield; Essential oil content
1. Introduction
Indiscriminate use of chemicals and fertilizers has
altered the biological ecosystem, affected non-target
organisms and adversely influenced microorganisms in
the soil. Organic farming, which aims at cultivating theland and raising crops in such a way to keep the soil
alive and in good health may be an alternative to the
present system of farming which solely depends on
chemicals (Dahama, 1996).
Cymbopogon winterianus Jowitt (Java citronella) is an
important essential oil yielding aromatic grass cultivated
in India mainly in the lower hills of Assam, Karnataka
and Southern Gujarat. The steam volatile essential oilsextracted from its leaves are used in perfumery, cosmet-
ics, pharmaceuticals, and flavoring industries (Guenther,
1950; Wealth of India, 1985). Cymbopogon spp. are
known to be very hardy plants that can grow in soil types
*Corresponding author. Fax: +91-11-2468-2144/2145.
E-mail address: [email protected] (A. Adholeya).
0960-8524/$ - see front matter � 2003 Elsevier Ltd. All rights reserved.
doi:10.1016/S0960-8524(03)00198-6
ranging from rich loam to poor laterite but with a pre-
ference to well drained and nutrient rich soils. These
plants are also known for their tolerance to soil salinity
and alkalinity to a higher level than other crops (Kak and
Kaul, 1997). Java citronella is known to respond signi-
ficantly to fertilizers. However, not much information isavailable regarding the cultivation of citronella in mar-
ginal soils amended with composts/manures.
Application of manure compost has a positive effect
on crop production (Woodbury, 1992) wherein the
nutrient deficiencies are correlated to the compost
addition. Improvement in the soil physical structure
after amendment with organic wastes such as municipal
solid waste, sewage sludge, and manure compost hasalso been well documented (Guisquiani et al., 1995;
Wong and Ho, 1991). However, high doses of manure
might be toxic to plants, animals and human beings
(Meek, 1974; Donahue, 1977) and thus, due caution
must be exercised to keep its detrimental effects in check.
The benefits of application of compost manures on crop
production (Wong et al., 1999) and enhanced recovery
of AM population (Gaur and Adholeya, 2000) have
312 Tanu et al. / Bioresource Technology 92 (2004) 311–319
been reported to be directly related to the rate of applied
dose. The effect of the extent and type of manure com-
posts on native AM inoculum potential applied for en-
hanced crop production has not been investigated under
field conditions where availability of major nutrients
and high P fixation is a limiting factor (Bhatia et al.,
1998) such as in north-western India.
We hypothesized that the plant productivity can beimproved in nutrient deficient soils supplemented with
organic amendments. The present investigation was
therefore designed under field conditions in a nutrient
deficient soil to (i) optimize productivity by varying the
dose and type of compost manures, and its effect on (ii)
the infectivity potential of native arbuscular mycorrhizal
fungi (AMF).
2. Methods
2.1. Experimental design and preparation of field
The field experiment was conducted at The Energy
Research Institute’s field station at Gual Pahari in Hary-
ana state, (77�120E; 280�350N; 255 m above mean sea
level) India. The annual precipitation at this site is 500–
700 mm. The soil was sandy loam hyperthermic typic
Haplutalf (pH 7.38; N 0.012%); NaHCO3 extractable P
2.81 mg kg�1 (Olsen et al., 1954) and exchangeable K 103mgkg�1. The experiment was laid on 24 · 15 m plot,
planted with three varieties of citronella, applied with
four types of compost manure (leaf compost, poultry
manure, vegetable waste and sewage sludge) at four
doses forming various treatments. The treatments were
tested using a split plot completely randomized size in
each block and were replicated three times thereby con-
stituting 48 experimental plots per variety.The site was prepared by repeated ploughing and
planking in order to achieve fine tilth. Various types of
compostmanure combinationswere applied during the field
preparation. Random soil samples were taken from each
bed and analyzed for macro- and microelements (Table 2).
2.2. Planting germplasm and compost manure application
Slips of high yielding varieties of Citronella (C. win-
terianus)-Manjusha, Mandakini and Bio-13 were pro-
Table 1
Chemical characteristics of various composts
Composts pH (1:2.5
soil:H2O)
Electrical
conductivity (dS/m)
Available P
(ppm)
VC 7.60a 6.25 24.20
PM 7.91 8.88 38.50
LC 7.25 9.87 25.35
SSL 7.25 3.60 44.60
VC––vegetable compost; PM––poultry manure; LC––leaf compost; SSL––seaMeans are average of three replicates.
cured from Central Institute of Medicinal and Aromatic
Plants, Experiment Station, Pantnagar, Uttranchal
State, India.
Of the four types of compost used, vegetable compost
and leaf compost was prepared by aerobic decomposi-
tion at TERI’s field station, Gual Pahari. The compo-
sting bin (3 m · 3 m · 1 m) comprising of four units (1.5
m · 1.5 m · 1 m) made of bricks laid in such a mannerthat the walls had gaps for natural aeration. The com-
posts were ready in nine weeks. Poultry litter was pro-
cured from a near by poultry farm and was allowed to
mature for three weeks until the desired C:N ratio was
achieved (Table 1). Sewage sludge with 80% moisture
was procured from the Okhla Sewage Treatment Plant,
Municipal Corporation of Delhi, Government of Delhi,
India and was air-dried. The four doses (40, 80, 100 and120 t ha�1) of each compost manure were mixed thor-
oughly into the soil in each treatment bed before
transplantation. No additional synthetic chemical fer-
tilizer and pesticides were applied during the experiment.
The nutrient profile of the four composts/manures is
given in Table 1.
Slips of all the varieties were transplanted into the
treatment plots at a spacing of 20 cm plant · plant and30 cm row · row.
2.3. Growing conditions and measurements
Standard agronomic practices were followed includ-
ing regular hoeing, weeding (ICAR, 1992). Plants were
irrigated as necessary using overhead sprinklers and
were harvested after 120 days. Samples of the shoot and
soil were drawn from each bed and processed separately.Fresh shoot weight (herbage yield) was recorded and the
sample was then dried and processed for extraction of
essential oil.
The number of infectious propagules of native AMF
in soil core samples drawn from each treatment bed was
determined by infectivity potential (IP) bioassay (Gaur
et al., 1998). One hundred grams of core samples were
put in plastic pots (7 cm height and 5 cm diameter) witheight germinated seeds of Sorghum vulgare. The pots
were kept in a greenhouse (28/22 �C, 68% RH), watered
with deionized water to a moisture content of 60% water
holding capacity and harvested after 14 days. The entire
K
(ppm)
Organic carbon
(%)
Total N
(%)
C:N
557.30 4.33 0.53 8.1
998.20 5.38 0.43 12.5
223.33 6.79 0.51 13.3
261.00 4.45 0.32 13.9
wage sludge.
Table 2
Chemical characteristics of experimental site/trial after the application of various composts
Amendments Dosesa pH (1:2.5) Electrical
conductivity
(dS/m)
Total N (%) Available P
(ppm)
Available K
(ppm)
O:C (%)
VC 1 7.65 0.27 0.024 12.40 105.20 1.23
2 7.65 0.21 0.033 18.86 160.80 1.91
3 7.63 0.31 0.034 22.22 196.60 1.85
4 7.56 0.24 0.035 26.60 210.00 2.20
PM 1 7.43 0.29 0.013 18.40 250.67 1.43
2 7.43 0.23 0.019 24.60 340.01 2.06
3 7.14 0.23 0.025 30.80 390.20 2.06
4 7.36 0.26 0.031 34.62 459.00 2.36
LC 1 7.43 0.36 0.010 10.98 123.30 0.83
2 7.57 0.42 0.015 14.40 256.67 1.64
3 7.57 0.36 0.020 17.60 286.00 1.99
4 7.42 0.48 0.023 23.28 303.66 2.14
SSL 1 7.30 0.80 0.011 13.80 117.20 0.76
2 7.34 0.76 0.019 19.64 140.33 1.60
3 7.38 0.56 0.022 28.09 174.00 1.71
4 7.65 0.70 0.028 32.50 191.30 2.01
VC––vegetable compost; PM––poultry manure; LC––leaf compost; SSL––sewage sludge.a Compost doses 1, 2, 3, and 4 are 40, 60, 100 and 120 t ha�1 respectively. Means are average of three replicates.
Tanu et al. / Bioresource Technology 92 (2004) 311–319 313
root system from each pot was collected, washed and
stained (Phillips and Hayman, 1970) for enumerating
the total number of entry points. The infection units
were finally expressed in IP g soil�1.The phosphorus content was determined by the
method of Kitson and Mellon (1944), after the wet
digestion of the plant samples. Essential oil content in
dried leaves was estimated using Clevenger’s apparatus
(Buggle et al., 1999).
Data were analyzed using analysis of variance (SAS
Institute Inc., 1991). Duncan’s least significant differ-
ences (LSD) was used to separate treatment means whenF -values were significant, using Costat Statistical soft-
ware (Cohort, Berkeley, CA).
3. Results and discussion
3.1. Plant growth and essential oil content
The results of the present study demonstrates the
response of three varieties of Java citronella grown in a
nutrient deficient soil amended with various types and
dose of compost manures. Plant growth in terms ofherbage, essential oil and nutrient uptake was recorded
in the various treatments. The study also reflected the
effect of these organic amendments on the native AM
fungi. Out of the four different organic amendments,
PM applied at 100 t ha�1 was found to be the optimum
dose for maximum herbage yield in all the three varieties
of Java citronella. However, amongst the three varieties,
Bio-13 was found to produce the maximum herbage
yield. Although higher herbage yield was also recorded
in the 120 t ha�1 PM amended soils, the yield was found
to be non-significantly higher. This may imply that the
nutrient requirement of the plants has been fulfilled at100 t ha�1 PM amendment and further increase in the
rate of applied manure was not necessary. Improved
growth in plants has also been reported by Obi and Ebo
(1995), where the authors reported improvement in the
yield of maize when grown in severely degraded sandy
soils that were amended with PM. The present study
also reveals the effect of other types of organic amend-
ments on the growth of Java citronella. Regressionequations (Fig. 1) were developed on the basis of the
relationship between the various compost/manure dose
and herbage yield in the given nutrient deficient soil. The
results revealed that the correlation coefficient of the
Java citronella varieties varied with the type of compost/
manure amendment. Among the three varieties, Bio-13
showed the highest herbage yield (10.57 t ha�1) followed
by Mandakini (10.38 t ha�1) and then Manjusha (10.05t ha�1) at 100 t ha�1 PM application (Fig. 1). The
essential oil content (Figs. 2–4) was also found to follow
the same pattern as observed in herbage yield. Signifi-
cantly higher essential oil content was recorded in the
plants grown in the PM amended plots when compared
to the plants grown in the other organic amendments.
Amongst the varieties, highest essential oil content was
recorded in the Bio-13 variety grown at 100 t ha�1 PM.The response of Java citronella may also be attrib-
uted due to the increased total organic matter, macro-
and micronutrients rendered after the application of
various compost/manure (Table 2). The soil organic
0
6
8
10
12 VC PM LC SSL Regression
0
6
8
10
12
Compost doses (t ha-1)0 20 40 60 80 100 120 140 160
0
6
8
10
12
Manjusha
Mandakini
Bio-13
VC, y* = 5.34 + 0.0369 x**, r2= 0.979 PM, y = 6.05 + 0.0366 x, r2= 0.966LC, y = 5.56 + 0.0301 x, r2= 0.902SSL, y = 5.63 + 0.0385 x, r2= 0.872
VC, y* = 6.21 + 0.0351 x**, r2= 0.968 PM, y = 6.12 + 0.0382 x, r2= 0.935LC, y = 5.19 + 0.0409 x, r2= 0.962SSL, y = 6.10 + 0.0367 x, r2= 0.970
VC, y* = 6.01 + 0.0315 x**, r2= 0.922 PM, y = 7.36 + 0.0287 x, r2= 0.936LC, y = 5.50 + 0.0340 x, r2= 0.954SSL, y = 5.90 + 0.0346 x, r2= 0.979
Her
bage
yie
ld (t
ha-1
)H
erba
ge y
ield
(t h
a-1)
Fig. 1. Influence of various compost manures and its relationship with herbage yield on three varieties of Cymbopogon winterianus: y� ¼herbage
yield; x�� ¼ compost dose.
314 Tanu et al. / Bioresource Technology 92 (2004) 311–319
matter and macronutrients concentration was found to
increase as the rate of applied organic amendment in-
creased. Similar results have also been reported by
Wong et al. (1999) on amending soils with manure
composts. However, the authors considered single
manure compost while this study involves different types
of organic manures/composts. The authors reportedincreased plant dry weight due to the better nutrient
status of N, P and K in the soils as well as in the plants.
The increased plant yield may also be due to increased
rhizosphere aggregate stability which might have fa-
vored the beneficial microbes which in turn could have
contributed to improved biomass (Caravaca et al.,
2002). In the present study, of all the amendments, PM
at 100 t ha�1 resulted in the maximum herbage yield
as well as higher P uptake in all the three varieties of
citronella.
3.2. Effect of organic amendment on native AM popula-
tions and root colonization
The type of organic amendment had a significant
effect upon the indigenous populations of mycorrhizawith respect to their infectivity potential (IP). Overall,
there was an increase in the native AM population in
soils amended with LC in all the three-citronella varie-
Dry
wei
ght (
g/pl
ant)
0
5
10
15
20
25
Compost dose (t ha-1)
Plan
t P (p
pm)
0
500
1000
1500
2000
2500
LSD (0.05%) Type = 0.39 Dose = 0.46
LSD (0.05%) Type = 50.36 Dose = 69.52
40 80 100 120
0
50
100
150
200
250VC PM LC SSL
LSD (0.05%) Type = 3.99 Dose = 4.01
Oil
yiel
d (
l pl
ant-1
)µ
Fig. 2. Essential oil content, shoot dry weight and P content in plant tissues of Cymbopogon winterianus var. Manjusha influenced due to amendment
of different types of compost manures applied at various doses in an Indian alfisol. The treatment means were separated by two-way ANOVA
through DMRT. LSD––least significant difference (p ¼ 0:05) by DMRT.
Tanu et al. / Bioresource Technology 92 (2004) 311–319 315
ties. This is concurrent to the report by Baby and
Manibhushanrao (1996) where leaf manure amendments
stimulated high arbuscule development and sporulation
in a rice crop. The 40 t ha�1 SSL treated plots did not
show significant increase in AM infective potential for
all varieties when compared to plants grown in other
composts/manures. In general, the infectivity potential
of soils showed a linear increase with increase in dose of
compost manures (Fig. 5). However, the maximum in-
crease in native mycorrhizal population in terms of
infectivity potential was recorded in variety Bio-13 (6.6
IP/100 g soil) in LC plots amended with 120 t ha�1 (Fig.
5). The least IP was recorded in Mandakini (0.6 IP/100 g
soil) at 40 t ha�1 amendment of SSL (Fig. 5). The
0
5
10
15
20
25
Compost dose (t ha-1)
0
500
1000
1500
2000
2500
LSD (0.05%) Type = 0.51 Dose = 0.72
LSD (0.05%) Type = 63.87 Dose = 52.19
40 80 100 120
0
50
100
150
200VC PM LC SSL
LSD (0.05%) Type = 4.72 Dose = 6.25
Dry
wei
ght (
g/pl
ant)
Plan
t P (p
pm)
Oil
yiel
d (
l pl
ant-1
)µ
Fig. 3. Essential oil content, shoot dry weight and P content in plant tissues of Cymbopogon winterianus var. Mandakini influenced due to
amendment of different types of compost manures applied at various doses in an Indian alfisol. The treatment means were separated by two-way
ANOVA through DMRT. LSD––least significant difference (p ¼ 0:05) by DMRT.
316 Tanu et al. / Bioresource Technology 92 (2004) 311–319
variation in the IP amongst the varieties as observed in
this study indicates host dependency and can be ex-
plained on the basis of numerous reports (Bever et al.,
1996; Daft and Hogarth, 1983; Hetrick and Bloom, 1986;Hung and Sylvia, 1988; Koomen et al., 1987) where the
authors described that the sporulation rates of AM fungi
were found to be host dependent under laboratory sys-
tems. There are also a number of reports on the effect of
organic amendment application on indigenous VAM
populations. Douds et al. (1997) reported an increase in
AM fungi spore populations of certain AM species in
soils receiving organic amendments in the form of
chicken litter/LC in comparison to those soils receivingraw diary manure and conventional fertilizers (M€aderet al., 2000). Verma and Arya (1998) reported an
enhancement of AM spore production by several times
in bamboo rhizosphere amended with organic manure.
The enhancement of the spore production in organic
0
500
1000
1500
2000
2500
Compost dose (t ha-1)
0
500
1000
1500
2000
2500
LSD (0.05%) Type = 0.68 Dose = 0.54
LSD (0.05%) Type = 31.64 Dose = 45.45
40 80 100 120
0
50
100
150
200
250 VC PM LC SSL
LSD (0.05%) Type = 6.13 Dose = 5.98
Dry
wei
ght (
g/pl
ant)
Plan
t P (p
pm)
Oil
yiel
d (
l pl
ant-1
)µ
Fig. 4. Essential oil content, shoot dry weight and P content in plant tissues of Cymbopogon winterianus var. Bio-13 influenced due to amendment of
different types of compost manures applied at various doses in an Indian alfisol. The treatment means were separated by two-way ANOVA through
DMRT. LSD––least significant difference (p ¼ 0:05) by DMRT.
Tanu et al. / Bioresource Technology 92 (2004) 311–319 317
manure amended medium may be due to enhanced
growth and spread of AM hyphae as reported by Hep-
per and Warner (1983), Joner and Jakobson (1992) and
St John et al. (1983). Muthukumar and Udaiyan (2002)
while working on mycorrhizal response of cowpea under
organic farming reported higher mycorrhizal production
in plants grown in soils amended with organic manure
than in the non-amended soils. They also reported thatthe mycorrhizal response highly correlated with the type
of organic manure. They also suggested that the growth
and yield of cowpea in response to organic amendment
was a result of the differential effects of the types of
amendment on the production of different AM fungi
within the community.
The lower IP of the native AM fungi in SSL and PM
amendments could be due to the high available P in
these soils. PM is known to increase the availability of Pin soils (Toor and Bahl, 1997) and hence a negative effect
No.
of i
nfec
tious
pro
pagu
les
(per
100
g s
oil)
0
1
2
3
4
5
6 ManjushaMandakiniBio-13
0
1
2
3
4
5
6
7
0
1
2
3
4
5
6
7
8
Compost dose (t ha-1)
40 80 100 120 0
1
2
3
4
5
6
7
Vegetable compostLSD (0.05) Variety = 0.91 Compost dose = 1.25
Poultry manureLSD (0.05) Variety = 1.33 Compost dose = 1.03
Leaf compostLSD (0.05) Variety = 1.58 Compost dose = 1.48
Sewage sludgeLSD (0.05) Variety = 0.99 Compost dose = 1.27
No.
of i
nfec
tious
pro
pagu
les
(per
100
g s
oil)
Fig. 5. Effect of compost manure upon the native AM population in
the rhizosphere of three varieties of Cymbopogon winterianus.
318 Tanu et al. / Bioresource Technology 92 (2004) 311–319
on the native AM populations and enhanced herbage
yield and P uptake was observed in plants grown in this.
Higher soil P availability is known to suppress VAM
fungal colonization (Ryan and Ash, 1999; Sharma andAdholeya, 2001; Valentine et al., 2001) and spore den-
sity (Menge et al., 1978).
While comparing the nutrient profile of the experi-
mental site with the ratings of medium fertility levels
(Singh et al., 1999), the site was deficient in essential
major nutrients. The compost manure application at the
rate of 40 t ha�1 in the soil supplied these nutrients to
levels observed in medium fertility soils but the maxi-
mum response in terms of herbage yield has been
exhibited at the 100 t ha�1 PM amendment thus imply-ing the high levels of organic amendments required
(Table 2).
The regression equation models developed in the
present study can be used to assess the rate of compost/
manure amendment required to achieve the desired
herbage yield for citronella growers of a region. The
results of this study also strongly suggest the possibility
of obtaining increased citronella production under or-ganic agriculture by applying appropriate combinations
of organic amendments.
Acknowledgements
The present study was supported by funding from the
Department of Biotechnology, Government of India toAA. The authors wish to thank the Director General,
Dr. R.K. Pachauri, The Energy Research Institute for
providing the infrastructure facilities. The authors wish
to thank Dr. Mahaveer P. Sharma for providing intel-
ligent inputs during designing of experiment and statis-
tical advice. The authors also wish to acknowledge
Mr. U. Gangi Reddy for his help in the soil and plant
analysis. The technical assistance of Mr. SailendraKumar in conducting the experiment and subsequent
analysis work is duly acknowledged.
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