Journal of Essential Oil Bearing Plants

ISSN: 0972-060X (Print) 0976-5026 (Online) Journal homepage: https://www.tandfonline.com/loi/teop20

Influence of Harvesting Time on Essential Oil

Content, Chemical Composition and Pesticidal
Activity of Artemisia maritima Growing Wild in the
Cold Desert Region of Western Himalayas

Swati Walia, Arti Rana, Ashok Singh, Mohit Sharma, S.G. Eswara Reddy &
Rakesh Kumar

To cite this article: Swati Walia, Arti Rana, Ashok Singh, Mohit Sharma, S.G. Eswara Reddy
& Rakesh Kumar (2019) Influence of Harvesting Time on Essential Oil Content, Chemical
Composition and Pesticidal Activity of Artemisia�maritima Growing Wild in the Cold Desert
Region of Western Himalayas, Journal of Essential Oil Bearing Plants, 22:2, 396-407, DOI:
10.1080/0972060X.2019.1610077

To link to this article: https://doi.org/10.1080/0972060X.2019.1610077

Published online: 22 May 2019.

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 TEOP 22 (2) 2019 pp 396 - 407 396
ISSN Print: 0972-060X
ISSN Online: 0976-5026

Influence of Harvesting Time on Essential Oil Content, Chemical

Composition and Pesticidal Activity of Artemisia maritima Growing
Wild in the Cold Desert Region of Western Himalayas

Swati Walia 1,2, Arti Rana 2, Ashok Singh 4, Mohit Sharma 1,3
,
S.G. Eswara Reddy 1,2* and Rakesh Kumar 1,2*

Academy of Scientific and Innovative Research

1

Agrotechnology of Medicinal, Aromatic and Commercially Important Plants Division

2

3
Natural Product Chemistry and Process Development Division
4
High Altitude Biology Division, CSIR-Institute of Himalayan Bioresource Technology,
(Council of Scientific and Industrial Research), Post Box No. 6, Palampur 176 061 (H.P.), India
Received 02 November 2018; accepted in revised form 16 April 2019

Abstract: Essential oil (EO) of Artemisia maritima plants growing naturally in the cold desert region
has been extracted by hydro distillation at different growth stages during 2015-16. The constituents of the EO
have been characterized by gas chromatography (GC) and GC-mass spectrometry. Nineteen compounds were
identified. EO content and composition vary throughout the vegetation period of the plant. The best time for
higher EO content (0.45±0.15 %) is August, while for higher 1,8-cineole (59.24±1.63 %) is during October. EO of
A. maritima was characterized by high content of 1,8-cineole, bornyl acetate, terpinen-4-ol, camphor, sabinene
and germacrene D which varied from 90.54±7.76 to 92.41±6.50 % of the total oil content. The EO of A. maritima
was evaluated for their acaricidal activity against two-spotted spider mite (Tetranychus urticae) (Acarina:
Tetranychidae). EO reported excellent fumigant activity against T. urticae with LC50 value of 323.86, 27.05, 15.48
and 12.15 mgL-1 after 1, 2, 3 and 4 h of treatment, respectively as compared to positive control i.e. Propargite 57
% EC (755.72 mgL-1). In repellent activity, 10000 mgL-1 showed significantly higher repellency (95.83±1.67 %) to
adult T. urticae and was at par with positive control i.e. Azadirachtin 0.15 % EC (96.67±3.33 %) and 8000 mgL-1
(91.67±2.20 %) as compared to other concentrations studied.

Key words: Artemisia maritima, harvesting time, 1,8-cineole, Tetranychus urticae, fumigant.

Introduction mintic and very effective against round worms

Artemisia maritima L. is an aromatic perennial
herb commonly known as sea wormwood (fam-
ily Asteraceae). It is distributed in the western
Himalayas at altitudes of 7000-9000 feet and
grows abundantly in north-west Kashmir, dry tem-
perate region of Himachal Pradesh 1 and to a
certain extent in Chamoli district of Garhwal
Himalayas (Uttarakhand) 2. The plant is commer-
cially utilized in India for santonin which is anthel-
(Ascaris spp.). Distinctive parts of the plant have
been used for medicinal purposes; leaves for
asthma, stomach torment (antispasmodic) and
CNS ailments, while seeds are used as an appe-
tizer, expectorant, aphrodisiac, ophthalmia, a cure
for indigestion, diarrhoea, inflammation, a heart
stimulant and tonic 3. The essential oil of A. mar-
itima shows anti-infective, antibacterial, antifun-
gal, antispasmodic, antimalarial and bronchodila-

*Corresponding authors (S.G. Eswara Reddy; Rakesh Kumar)

E-mail: < ereddy2001@yahoo.com; rakeshkumar@ihbt.res.in > © 2019, Har Krishan Bhalla & Sons
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407
tory activities 1,4. The chemical composition of its
essential oil has been extensively studied. The
main compounds of the essential oil of A. mar-
itima are 1-8 cineole, camphor, borneol, bornyl
acetate and germacrene-D 5. However, the con-
tent of essential oils and its constituents reported
varying significantly depending upon variations in
climatic and agricultural conditions, growing sea-
son, growth/harvesting stage, plant parts, cultiva-
tion practices and origin 6.
The chemical composition of Artemisia spp.
from different origins show a dominant presence
of 1,8-cineole, α-thujone, β-thujone, davanone,
artemisian ketone, β-caryophyllene or caryo-
phyllene oxide, 1,8-cineole, camphor, and borneol
are the major compounds present in essential oil
A. maritima 7-9. The variation in the occurrence
of certain compounds in a plant is a function of
any one of, or a combination of, three factors:
genetically determined properties, the age of the
plant, and the environment. Ideally, an experiment
evaluating the relative importance of these fac-
tors should be designed with randomized blocks
and plants of equal age, collected by careful sam-
pling in a random and systematic manner from
the natural populations. In several species of
Lamiaceae, at flowering, essential oil is at the high-
est level, while in other species, flowering has a
lower influence 10. The present investigation deals
with the variations of yield and composition of the
essential oil obtained at different growth times by
plants collected from the same stand to limit the
influence of the factors previously cited.
Botanical formulations have received much im-
portance in organic farming/sustainable manage-
ment of insect pests due to low toxicity to natural
enemies of insects and non-target organisms. Due
to multiple modes of action, plant products are
gaining more attention as an alternative method
of control. Two-spotted spider mite (Tetranychus
urticae Koch) (Acarina: Tetranychidae) is an
important pest of different cultivated crop plants
grown under protected and field conditions caus-
ing losses in most years. Adults and nymphs suck
the sap from the lower surface of leaves which
results in yellowing and discoloration 11-13. In se-
vere incidence, the mites construct the spider webs
on the leaves and stem which results in drying
397
and dropping of leaves 14,15. Tetranychus urticae
was developed resistance to different acaricides
due to the non-judicious application and short life
cycle. The EOs reported toxicity, antifeedant, re-
pellent and ovicidal activities against different
pests 16,17 including T. urticae 18-20. Although, aca-
ricidal properties of different EOs have been re-
ported on T. urticae, but no report is available for
the EO of A. maritima. Therefore, the present
experiment was planned to study the fumigant and
repellent activity of A. maritima against T. urticae.
The population of A. maritima is abundantly
available in the barren land of Lahaul-Spiti and
Kinnaur district of Himachal Pradesh between
2700 m and 3400 m but the local people are not
making its effective use due to lack of knowledge
1
. Though local people use raw Artemisia plants
for insect repellency in the fields but they are un-
aware of its insecticidal property which can be of
great use in industries and can lead to good eco-
nomic returns. Many studies have been conducted
to determine essential oil content and composi-
tion of A. maritima, but there is no information
regarding its harvesting time and insecticidal ac-
tivity of its essential oil. Hence this study was
conducted to find out the effect of harvesting time
on essential oil content and compounds of A. mar-
itima from its natural population grown in cold
desert region of western Himalayas to obtain the
highest essential oil yield and to study its insecti-
cidal properties for industrial use.
Materials and methods
Experimental site
The experiment was performed at the research
center of CSIR-Institute of Himalayan Biore-
source Technology, Palampur (HP) India, Centre
for High Altitude Biology, CeHAB situated at
Tandi, Keylong Distt. Lahaul and Spiti, HP (3450
m amsl, 32°33’58.8" N, 76°58’21.8" E), during
2015-2016. Weather data (maximum and minimum
temperature, rainfall, soil temperature, and rela-
tive humidity) were obtained from weather tower
situated in CeHAB Ribling farm, Lahaul and Spiti,
HP (Figure 1). Weekly average data was recorded
during different harvesting time which was pooled
for 2015-16. In 2015-2016, the maximum tempera-
ture ranged from 17.34 to 21.46°C, whereas mini-
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407
mum temperature ranged from 4.02 to 11.09°C
during the harvesting period. Likewise, average
soil temperature ranged from 15.16 to 18.51°C,
mean relative humidity ranged between 38.56 to
68.80 % and 62.23 mm total rainfall was received
during the experimental period.
Plant material
Fresh plant material of A. maritima was col-
lected from Lahaul valley (latitude 32o.57 1 N and
longitude 77o.04 1 E) of Himachal Pradesh (In-
dia). The plant material was authenticated, and a
voucher specimen was deposited in the herbarium
(PLP 17794) of CSIR-IHBT, Palampur.
Experimental details
A. maritima fresh plant samples were harvested
from natural population during the fourth week, in
the afternoon, in different times of the growing
phase of the plant at the monthly interval i.e. from
June to October (during 2015 and 2016). In each
month 500 g of fresh aerial biomass viz. stem and
leaves were used for distillation. Material was
chopped and hydro-distillated in Clevenger appa-
ratus for four hours. Three replicates were used
for evaluating essential oil content and two repli-
cates for the composition of the oil. The experi-
ment was conducted in a randomized complete
block design. Essential oil content was measured
as mL and % ratios (v/w), as per procedure given
in European Pharmacopoeia 21. Extracted oil
samples were dried over anhydrous sodium sul-
phate, and stored at 4°C until analysis. Essential
oil chemical composition was analyzed in Natural
Product Chemistry and Process Development di-
vision of the Institute.
Identification of compounds through Gas
Chromatography (GC) and Gas Chromato-
graphy-Mass Spectrometry (GCMS)
Essential oil composition of A. maritima was
analyzed by Shimadzu GC-2010 gas chromato-
graph (Shimadzu, Tokyo, Japan) equipped with
flame ionized detector (FID) and ZB- 5MS (5 %
diphenyl, 95 % methyl polysiloxane stationary
phase) capillary column (30 m x 0.25 mm, 0.25
μm film thickness) from SGE International,
Ringwood, Australia. Carrier gas used was nitro-
398
gen with 1.05 mL min-1 flow rate. Oven tempera-
ture was programmed at 70°C fo 4 min and then
increased by 4°C min-1 to 220°C, with maintained
temperature of injector (240°C) and detector
(250°C).
Gas chromatography-mass spectrometry analy-
sis was carried out by Shimadzu QP2010 GC-
MS with AOC-5000 Auto injector and ZB- 5MS
capillary column (30 m x 0.25 mm, 0.25 μm film
thickness) in which nitrogen with 1.05 mL min-1
flow rate was used as carrier gas. Sample injec-
tion of 2 μL with split ratio of 10 and mass scan
40-800 amu. Injector and ion source temperature
were maintained at 240°C and 250°C, respectively.
At initial oven temperature used was 70°C for 4
min with a rising rate of 4°C min-1 upto 220°C
and then held isothermal at 220°C for 5 min 22.
Compounds were identified on the basis of reten-
tion indices (calculated using homologous series
of n-alkanes from C8 to C24) and peak area chro-
matogram. Components of oil were then identi-
fied by comparing retention indices (RI) values,
mass spectra and NIST (National Institute of
Standards and Technology) library 23,24.
Test mites
Tetranychus urticae used for the study was
collected from the field and reared on Phaseolus
vulgaris at 25±2°C, 60±5 % relative humidity and
L16:D8 h of photoperiod under laboratory for more
than 30 generations.
Acaricidal activities of essential oils against
T. urticae
Fumigant toxicity
Fumigant toxicity of the EO of A. maritima
against adult T. urticae was evaluated as per the
method described by Mozaffari et al. 18 with little
modification by Reddy et al. 17. Five concentra-
tions of essential oil collected in the month of
August (100, 50, 40, 20 and 10 mg L–1 volume by
air) were prepared.
P. vulgaris leaf discs (3 cm diameter) were
placed in Petri dishes (60 ml capacity) lined with
water-saturated cotton wool. With a fine camel-
hair brush, 10 adult mites were released on the
surface of the discs. Each concentration was rep-
licated thrice. Filter paper disks (Whatman No.
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407
1) cut into 2 cm in diameter and placed on the top
of petri dishes with cello tape and impregnated
with five concentrations of EO. The commercially
available acaricide Propargite 57 % EC (Omite®)
available in the market at recommended dose was
used for the control of T. urticae and other mite
species was used as a positive control for com-
parison. Observations on number of dead mites
were recorded at hourly interval up to 4 hours.
Repellent activity
Repellent activity of EOs was tested as per the
method followed by Kogan & Goeden 25 with little
modification by Reddy et al. 15. Briefly, 100 mg
of EO was dissolved in 10 mL of distilled water
containing 0.05 % Triton-X100 LR spreader and
sonicated for complete dissolution. Five concen-
trations of test solutions (10000, 8000, 4000, 2000
and 1000 mgL-1) were prepared from stock solu-
tions from the higher concentration and poured in
glass petri dishes. Fresh P. vulgaris leaf discs (3
cm diameter) were prepared from P. vulgaris
leaves. Half of the leaf disk was immersed into a
test solution for five seconds, and after drying,
the other half of the disc was immersed in water,
which served as control. After drying, leaf discs
were placed in Petri dishes lined with water-satu-
rated cotton wool. With a fine camel-hair brush,
10 adult mites were released on the leaf discs.
There were seven treatments and each treatment
was replicated three times. The commercially
available neem based formulations i.e. Neem Baan
(Azadirachtin 0.15 % EC at 5 ml L-1) and neem
oil at 5 ml L-1 available in the market at recom-
mended dose for the control of T. urticae and
other mite species was used as a positive control
for comparison. Observations on the number of
mites present in each half of the leaf disc was
counted after 1 h. The percent repellence was
calculated according to the following formula:
% Repellency = (C-T/N)*100; Where, T = num-
ber of mites in the treatment, C = number of mites
in the control, N = Number of mites used in the
test.
Statistical analysis
Statistical analysis of data was done as per the
standard analysis of variance technique for the
399
design used, and treatment means were compared
at P < 0.05 level of significance. The fumigant
toxicity experiment was repeated for three times
and mean data were used for analysis. Fumigant
toxicity data from all bioassays were corrected
for mortality in the control using Abbott formula
26
. The median lethal concentration (LC50), me-
dian lethal time (LT50) and their corresponding 95
% confidence intervals, Chi-square and regres-
sion coefficients were determined following Probit
analysis 27 using statistical software, version 16.
The data on repellence was subjected to one way
ANOVA using statistical software, version 16 and
means were compared by Duncan Multiple Range
Test (DMRT).
Results and discussion
Essential oil content
The fresh samples of A. maritima on
hydrodistillation yielded pale yellow oil with pleas-
ant odor. Essential oil content varied from 0.25 to
0.45 % on the weight of fresh leaf material in the
season that lasts from June to October. Signifi-
cantly higher essential oil content was obtained in
August as compared to other months. This may
be because of the reason that flowering in A.
maritima occurs during July-August, also these
months recorded maximum temperature, rainfall
and relative humidity (Figure 1) which do not pro-
duce sufficient IAA for cell division and leads to
flower formation 28, due to which highest essen-
tial oil content is reported during this period. Kaul
et al. 29 also reported low oil content in summer
when the day temperature cross 40°C and high
oil content during moderate rainfall with high rela-
tive humidity in rose-scented geranium. The
present result showed that flowering stage i.e.,
during July-August is best suitable harvesting stage
for A. maritima, a further delay of harvest de-
creased the essential oil content of A. maritima.
An earlier study in Garhwal region of Uttara-
khand, also reported higher essential oil content
in Artemisia when harvested during the third
week of August 30. Likewise, Sharma et al. 5 in
A. maritima reported 0.33 % essential oil content
when harvested in the month of July in the Cold
Desert region of Himachal Pradesh. However,
Sah et al. 9 reported opposite results with higher
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407
Figure 1. Monthly mean weather data (temperature, rainfall , and relative humidity)
during the harvesting season at Lahaul &Spiti, India. RH, Relative humidity
(0.50 %) essential oil content in A. maritima with
late harvesting during the month of November.
Essential oil composition
Fifteen components were determined and iden-
tified by GC and GC-MS, representing about 77.43
to 88.12 % of the essential oil of A. maritima
(Table 1). Comparison of the analytical data of
the oils revealed marked differences in qualita-
tive and quantitative composition. The major con-
stituents of the essential oil were 1,8-cineole,
bornyl acetate, terpinen-4-ol, camphor, sabinene,
germacrene D, etc. The essential oil of A. mar-
itima possessed the highest amount of 1,8-cin-
eole during all the harvesting stages. These re-
sults are in accordance with the reports of Jaitak
et al. 31 and Sharma et al. 5 on A. maritima. The
altitudinal impact was exhibited by an investiga-
tion of Jaitak et al. 31 where plants collected from
different regions of western Himalaya revealed
distinctive percentage of 1,8-cineole with 23.8 %
from Pooh area (2500 m), 37.3 % from Rhongtong
pass (3500 m) and 44.2 % from Lahaul & Spiti
(4500 m) district of HP. Generally, the composi-
tion of A. maritima essential oil varied between
investigations published in the literature. Some
older publications reported α-thujone (63.3 %) 7
400
or camphor (44.3-44.4 %) as major constituents
32
. Sah et al. 9 identified chrysanthenone (25.7 %)
as the main compound in A. maritima EO. These
data illustrate the existence of different
chemotypes that may be due to differences in lo-
cation, soil type, altitude, and physiological stage
of development. The variation in certain com-
pounds in a plant is due to genetic makeup, the
age of the plant and the environment.
Among different harvesting time, harvesting in
the month of October recorded a significantly
higher percentage of 1,8-cineole which was 28.47
% higher than August month which recorded low-
est (Figure 2). 1,8-cineole was reported maximum
in October followed by July>June>
September>August. Each compound had a unique
profile for accumulation as a function of tempera-
ture and precipitation 33. As October month re-
corded lowest temperature and rainfall (Figure 1),
so it can be assumed that maximum accumula-
tion of 1,8-cineole occurred at low mean tempera-
tures (between 10-18°C), and at low precipita-
tion level. Significantly higher percentage of Cam-
phene and bicyclogermacrene were also reported
in the month of October and lowest in June. Bornyl
acetate and Terpinen-4-ol was found significantly
higher in the month of June, which was 67.95 %
 Table 1. Effect of different harvesting times on essential oil content and
composition (v/w %) of A. maritima (pooled data of 2015 and 2016)
Total

Borneol

Linalool
Camphor

Sabinene

Camphene
1,8-Cineole

β-Myrecene
Phellandrene
Terpinen-4-ol
Germacrene D

Bornyl acetate
Sabinyl acetate

Santolina triene
Bicyclogermacrene

Essential oil (v/w %)

Chrysanthenyl acetate

Litt. RI 908 953 976 991 1031 1033 1098 1143 1165 1177 1262 1285 1291 1480 1494
Exp. RI 902 950 973 990 1024 1033 1098 1146 1161 1180 1256 1283 1286 1479 1493
June 0.37c 0.69c 0.87de 2.68a 1.57cde 1.68bcd 41.28bc 4.31a 1.78d 0.99cde 14.20a 2.03a 13.47a 0.58cde 1.14abc 0.33cd 88.12ab
July 0.41b 4.88a 0.99d 2.14bc 2.72bc 2.53ab 43.09b 0.84bcd 3.87b 1.11cd 11.39b 1.04bc 8.10d 2.05a 0.95bcd 0.38c 86.39abc
August 0.45a 0.44d 2.16ab 2.52b 2.18bcd 2.82a 38.87de 0.86bc 5.52a 1.28bc 9.71bcd 1.19b 13.01abc 1.23bc 1.44a 0.65b 84.25abd
e
September 0.33d 3.87ab 1.34c 0.81e 5.81a 1.33ce 39.02cd 1.23b 3.43bc 3.66a 11.16bc 1.19b 13.15ab 1.47ab 0.39 0.07e 88.14a
ab
October 0.25e 0.00e 2.44a 1.29d 3.12b 2.05bc 49.94a 0.52bcde 1.75de 2.68ab 9.69bcde 0.90cd 8.02de 0.63cd 1.36 1.11a 77.43e
SEm ± 0.004 0.49 0.18 0.20 0.53 0.31 0.75 0.28 0.38 0.49 0.89 0.17 1.40 0.26 0.16 0.09 2.26
Swati Walia et al., / TEOP 22 (2) 2019 396 - 407

LSD (<0.05) 0.013 1.46 0.54 0.59 1.58 0.92 2.23 0.84 1.13 1.46 2.63 0.52 4.16 0.77 0.46 0.25 6.73

RI, retention indices; SEm±, Standard Error Mean; LSD, Least Significant Difference
401
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407
Intensity
min.
Figure 2. GC chromatograms of essential oil of
Artemisia maritima harvested in the month of October
and 46.54 % higher, respectively, as compared to
October which recorded the lowest value. A sig-
nificantly higher value of Camphor and germa-
crene D was reported in the month of August,
which further decreases with delaying time and
reported 215.42 % and 269.23 % higher as com-
pared to October and September, respectively.
Sabinene, linalool and chrysanthenyl acetate were
reported significantly higher in June which de-
creases with delaying harvesting time or say ma-
turity, while the opposite was seen in case of
Borneol and β-myrcene which significantly in-
creased in September with the lowest value in
the month of June. Other compounds of essential
oil did not show any consistent trend over the har-
vesting time. Previous studies have shown that
changes in the oil composition as a result of growth
development are dependent on the species and
the nature of compounds. Some compounds are
increased with the age of plants while some de-
creases. Chauhan et al. 34 found that thymol re-
corded its maximum amount in Origanum vulgare
at the late vegetative stage, while for γ-terpinene,
the maximum % was recorded at the reproduc-
tive stages (flower initiating, full blooming, and fruit
set). Letchamo et al. 35 also reported that in
402
Valeriana officinalis, major compounds viz.
valerenal, valerenic acid, and α-humulene con-
tents increased with the age of plants. Since 1,8-
cineole is the major and most important constitu-
ent of A. maritima essential oil, so to get its maxi-
mum percentage plant should be harvested in
October.
Acaricidal activity of EO of A. maritima
Fumigant toxicity of EO of A. maritima against
T. urticae
The fumigant toxicity of EO of A. maritima
against adults of T. urticae for dose-response bio-
assay to calculate the LC50 and LT50 to kill 50 %
of the test insect relative to control values at dif-
ferent hours after treatment was presented in
Table 2 and Table 3. Results showed that, all the
concentrations of EO showed toxicity to T.
urticae after treatment. LC50 values of EO at 1,
2, 3 and 4 h after treatment was 323.86, 27.05,
15.48 and 12.15 mgL-1, respectively. Similarly, LT
50 to kill 50 % of test mite at 100, 50, 40, 20 and
10 mgL-1 was 1.35, 1.58, 1.61, 2.71 and 3.94 h,
respectively as compared to positive control i.e.
Propargite 57 % EC (755.72 mgL-1). The toxicity
of A. maritima oil against T. urticae varies with
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407 403
Table 2. LC50 values for the fumigant toxicity of essential
oil of A. maritima against T. urticae (HAT)

Time (HAT) LC50 (mgL-1) 95 % Cl (mgL-1) Slope ± SE Chi square P value

1h 323.86 96.41-417.50 0.92 ± 0.36 1.02 0.80

2h 27.05 12.72-71.29 0.99 ± 0.31 0.19 0.98
3h 15.48 8.98-21.32 1.71 ± 0.35 1.30 0.73
4h 12.15 7.80-15.90 2.45 ± 0.45 1.35 0.72
Propargite 57 % EC (24 h) 755.72 95.68-1511.05 1.32 ± 0.37 1.24 0.74

HAT: Hours after treatment

Cl: Confidence limits

Table 3. LT50 values for the fumigant toxicity of

essential oil of A.maritima against T. urticae

Concn. LT50 (h) 95 % Cl (h) Slope ± SE Chi square P value

100 mgL-1 1.35 1.06 - 1.60 3.98 ± 0.68 1.61 0.40

50 mgL-1 1.58 1.24 - 1.88 3.44 ± 0.61 0.35 0.84
40 mgL-1 1.61 1.23 - 1.94 3.13 ± 0.59 0.06 0.97
20 mgL-1 2.71 2.02 - 4.28 1.87 ± 0.55 0.27 0.87
10 mgL-1 3.94 2.98 - 7.90 2.17 ± 0.62 1.32 0.52

h- Hours after treatment

Cl- Confidence limits
time after treatment/spray. Initially the mortality
will be slow and then gradually increased and
constant after some time because these EOs take
time to act on insect as compared to synthetic
pesticides. Similarly, the higher dose of EO took
less time to kill the 50 % of mite population as
compared to lower concentrations (Table 2). Use
of higher dose of EO (100 mgL-1), the air would
very probably be saturated with fumes of EO much
more quickly, thereby attaining more mortality.
Volatile nature of EO plays an important role
for the toxicity, however, not essentially all the
constituents of the EO have the same degree of
volatility/mode of action. The chemical constitu-
ents’ of EO may vary depending upon genotype,
soil, climatic conditions as well as season and ex-
traction method 36. The toxicity of the EO is also
varied between plant species, chemical composi-
tion and proportion of the constituents. The in-
secticidal activities of EOs are may be due to the
major chemical constituents but not always 37.
In the present study, EO of A. Maritima
showed good fumigant toxicity to adult T. urticae.
Our results conform with the results of others who
reported the fumigant toxicity of other EOs to T.
urticae. Piper aduncum and Thuja orientalis
oil showed good fumigant activity to T. urticae
38,39
. In a similar experiment, Eucalyptus
codonocarpa, and E. dives oil (10 μl L-1 air)
showed excellent fumigant action against T.
urticae 40. Similarly, EO of Cuminum cyminum,
Syzygium aromaticum, Mentha spicata, Achil-
lea mellifolium and A. filipendulina also reported
fumigant activity to T. urticae 41-43.
Repellent activity of EO of A. maritima
against T. urticae
Repellent activity of EO of A. maritima was
evaluated against T. urticae and results were pre-
sented in Table 4. Among the tested concentra-
tions, A. maritima at 10000 mgL-1 showed signi-
ficantly (one way ANOVA; F 6, 20 = 42.74;
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407
Table 4. Repellent activity of essential oil of A. maritima oil against T. urticae
Concn. (mgL-1)
10000
8000
4000
2000
1000
Azadirachtin 0.15 % EC
Neem oil 0.50 %
F value F6, 20 = 42.74; p< 0.0001
Means followed by the same letters within a column do not differ significantly by DMRT
p<0.0001) higher repellency (95.83±1.67 %) to
adult T. urticae and was at par with positive con-
trol i.e. Azadirachtin 0.15 % EC (96.67±3.33 %)
and A. maritima oil at 8000 mgL-1 (91.67±2.20
%) followed by 4000 mgL-1 (61.67±3.0 %) and
2000 mgL-1 (47.50±5.0 %). Similarly, the other
positive control (Neem oil 0.50 %) which is used
for the control of T. urticae in greenhouse and
field conditions is inferior to higher concentration
of A. maritima oil. Most of the EO and its com-
pounds presently used as repellents. The monot-
erpenes (eg., limonene, cineole, eugenol, citronel-
lal, etc.) present in the EOs are may be mainly
responsible for repellent activity of T. urticae. In
the present study, EO of A. maritima showed
excellent repellent activity to T. urticae. Present
results are in agreement with the reports of other
researchers who reported repellent activity to T.
urticae. The EO from Protium bahianum Daly
44
, Piper aduncum L. 38, Callistemon sieberi, E.
bicostata, E. ovata, E. sideroxylm, E. manni-
fera, E. dives, E. elata, E. condonocarpa,
Kunzea ericoides, Melaleuca armillaris and M.
fulgens 19, M. pulegium 18 , Elettaria carda-
momum L 45, Lavandula x intermedia Emeric
ex Loisel, Hyssopus officinalis 46, S. aromaticum
and M. Spicata 41 showed promising repellent
activity to T. urticae.
Conclusion
In general, it can be concluded that harvesting
time improves the essential oil content and oil com-
position of A. maritima as components of the es-
sential oil show fluctuations in their relative
404
% repellence
95.83 ± 1.67 a
91.67 ± 2.20 a
61.67 ± 3.00 c
47.50 ± 5.00 d
35.83 ± 3.63 e
96.67 ± 3.33 a
80.00 ± 5.77 b
amounts throughout the period under study. The
plant can be harvested in the month of July to
August for getting higher essential oil, however,
for higher 1,8 cineole the plant is to be harvested
in the month of October. Monoterpene 1,8-cin-
eole, bornyl acetate, terpinen-4-ol, camphor,
sabinene, and sesquiterpene germacrene D are
found to be alternatively the main components of
the oil: monoterpene before and after the flower-
ing and sesquiterpene before flowering. The EO
of A. maritima showed excellent fumigant and
repellent activity to T. urticae as compared to
positive controls i.e. Propargite, Azadirachtin/
neem oil. Production of any botanical formulation
requires continuous availability of plant material
on a commercial scale. The population of A. mar-
itima is abundantly available in barren land but
the local people are not making its effective use
due to lack of knowledge of processing techniques
and processing equipment. The standardization of
process technology for the extraction of essential
oil from plant parts and crude extracts of A. mar-
itima should be further evaluated against target
insect pests in the field. Based on field bio-effi-
cacy data and feasibility, the botanical formula-
tion may be recommended for the commercial
application in the greenhouse/field conditions for
the target pests.
Acknowledgments
The authors are grateful to the Director, CSIR-
IHBT, Palampur for necessary facilities during
the course of study. We also thank Mr. Shiv Kumar,
Senior Technical Officer for providing technical
 Swati Walia et al., / TEOP 22 (2) 2019 396 - 407 405
support during the work. Authors are also grate- nology, GOI, New Delhi, and Council of Scien-
ful to Dr. Bikram Singh, Chief Scientist (Retd.), tific and Industrial Research (CSIR) is also ac-
NPC&PD division for help in identification of knowledged. IHBT communication number for
essential oil compounds. Financial grant received this article is 4228.
from SERB, Department of Science and Tech-

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