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Elemental, biochemical and essential oil modulation in

developing seedlings of Psoralea corylifolia L. exposed
to different presowing gamma irradiation treatment
a

Sumira Jan , Talat Parween , Rehana Hamid , T.O. Siddiqi & Mahmooduzzafar
a

CORD, University of Kashmir, Srinagar, Jammu and Kashmir, India

IPFT, Gurgaon, India

Department of Botany, Jamia Hamdard, New Delhi, India

Published online: 30 Mar 2015.

To cite this article: Sumira Jan, Talat Parween, Rehana Hamid, T.O. Siddiqi & Mahmooduzzafar (2015): Elemental,
biochemical and essential oil modulation in developing seedlings of Psoralea corylifolia L. exposed to different presowing
gamma irradiation treatment, Journal of Essential Oil Research, DOI: 10.1080/10412905.2015.1024890
To link to this article: http://dx.doi.org/10.1080/10412905.2015.1024890

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Journal of Essential Oil Research, 2015

http://dx.doi.org/10.1080/10412905.2015.1024890

Elemental, biochemical and essential oil modulation in developing seedlings of Psoralea

corylifolia L. exposed to different presowing gamma irradiation treatment
Sumira Jana*, Talat Parweenb, Rehana Hamida, T.O. Siddiqic and Mahmooduzzafarc
a

CORD, University of Kashmir, Srinagar, Jammu and Kashmir, India; bIPFT, Gurgaon, India; cDepartment of Botany,
Jamia Hamdard, New Delhi, India

Downloaded by [University of Kashmir], [Sumira Jan] at 22:10 01 April 2015

(Received 6 March 2014; accepted 22 February 2015)

Seeds of Psoralea corylifolia L. were irradiated with different doses of gamma rays; 2.5, 5, 10, 15 and 20 kGy from
a cobalt source a at dose rate of 1.65 kGy/hour. The stimulatory effects of gamma irradiation at 10 kGy were evident
for biochemical and elemental parameters at the owering stage and thereafter declined. Nitrogen was found to be
maximum in leaves and sulfur in roots at the owering stage of seedlings grown from seeds exposed to 10 kGy.
Sulfur content was found to be most sensitive to gamma irradiation doses resulting in a maximum decline in stems
(89.10%), leaves (65.79%) and roots (57.07%) with 20 kGy, respectively. Amino acids exhibited a similar percent
decline (52.79%) at the pre-owering stage in plants raised from seeds irradiated with 20 kGy. Using gas
chromatographymass spectrometry (GCMS) and solid-phase particle microextraction (SPME) analysis, a remarkable percent increment was conrmed in tricyclene, -pinene, -myrcene, camphene, caryophyllene, -gurgenene,
n-nonanal, -(E)-ocimene, (Z)-3-hexene-ol and germacrene D in plants raised from seeds exposed to 20 kGy.
Maximum oil content (1.60%) was found in seeds irradiated with 20 kGy and minimum oil content was found in
control seeds (0.89%). The study demonstrates the inverse response between primary and secondary metabolites in
terms of enhancement in essential oil yield, as well as concentration of antioxidant components at 15 and 20 kGy.
Keywords: gamma irradiation; babchi seeds; essential oil content; volatile compounds; biochemical parameters

1. Introduction
In recent years, more attention has been paid to some
physical factors that favorably inuence the sowing
material of cultivated plants and improve disinfestation
of grains (13). The prevailing opinion is that physical
methods for the processing of pre-sowing material
(seed) stimulate metabolic changes in the seeds (47).
One of the physical methods that can be applied to the
improvement of the sowing material is seed treatment
with gamma radiation (8, 9). However, the viability
and sometimes the developmental process of the seedling or the plant have been seriously hampered by
radiation (10). The stimulatory effects of gamma rays
have been recorded on some aromatic plants such as
chamomile (11), peppermint (12, 13), lemongrass (14),
and Mathiola incana and Delphinium ajacis (15).
Psoralea corylifolia well known for its antioxidative, anti-microbial, anti inammatory, anti-tumor,
anti-mutagenic, anti-HIV activity, insect hormonal
activities and in the inhibition of mitochondrial lipid
peroxidation has been extensively used on a commercial scale by Indian pharmaceutical industries resulting
in fast decline of its wild population (1622). Apart
from active compounds, Psoralea corylifolia L. yields
a pale yellowish oil with a characteristic, terpenic,
*Corresponding author. Email: sumira.sam@gmail.com
2015 Taylor & Francis

powerful odor, consisted primarily of monoterpene,

sesquiterpene hydrocarbons and phenylpropaniod compounds (22). This oil is used externally for the treatment of leucoderma, psoriasis and leprosy in Indian
folk medicine (23). Traditional techniques for the mass
multiplication of P. corylifolia through seeds are unreliable due to its variable survival responses under natural
conditions (24). High root sensitivity and deliberate
seed germination further slows down the mass propagation of this plant (25, 26).
Hence, there is an urgent need for the development of
this valuable herb for large-scale production. To date,
there is no report on gamma irradiation used as a physical
elicitor to alter the secondary metabolism of P. corylifolia.
Therefore, better agro-technological methods are necessary if we are to increase metabolite yields. So far, no
efforts have been made to breed or select strains of certain
genotypes with higher harvest indices or increased level
of active ingredients. Little further information is available
for optimizing its cultivation and the harvesting of larger
seed crops. Thus, the present investigation was conducted
to tackle this issue by performing the metabolic and
phytochemical evaluation on Psoralea corylifolia L. after
exposure of seeds to variable doses of gamma rays for
optimizing the dose best for improving yield.

S. Jan et al.

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2. Experimental
2.1 Procurement of seeds and irradiation
Dry seeds of babchi [Psoralea corylifolia L.], belonging to the family Fabaceace, were acquired from
NBPGR (National Bureau Plant Genome Research),
Indian Agriculture Research Institute (IARI), New
Delhi. The seed samples (25 g each) were packed in
polythene and were irradiated with various doses of
gamma rays (T1=2.5, T2=5, T3=10, T4=15 and T5=20
kGy) from 60Co (Gamma chamber, GC, 5000) at a
dose rate of 1.65 kGy/hour (27) under ambient room
temperature in INMAS (Institute of Nuclear Medicine
and Applied Sciences), New Delhi.
Irradiated seeds were transferred and planted in the
herbal garden of Hamdard University, New Delhi, India
(2838N, 7711E; elevation 228 m), about 7080%
relative humidity, 35C temperature, sandy-loam soil
(pH 7.3). Soil analysis exhibited 51 and 7.9 ppm concentrations of nitrogen and sulfur levels, respectively.
Each sample was planted in six rows, 4 m long and 0.6
m wide, making an area of 14.4 m2. Hills were 30 cm
apart; ve seeds per hill to avoid root interruption in
Psoralea corylifolia L. owing to the high sensitivity of
roots. The leaves from developing seedlings were collected at three developmental stages [pre-owering (45
DAS), owering (90 DAS) and post-owering (135
DAS)] to analyze the effects of the gamma irradiation
on biochemical and elemental parameters. Field experiments were repeated three times with ve replicates.
The mature seeds of developing plants of different sets
(T0=control, T1=2.5, T2=5, T3=10, T4=15 and T5=20
kGy) were harvested in mid-November to estimate and
evaluate the oil yield and quality by gas chromatographymass spectrometry (GCMS) and solid-phase
particle microextraction (SPME) analysis.
2.2 Chemicals
All chemicals used were of analytical reagent (AR) or
guaranteed reagent (GR) quality. Most of the chemicals
were the products of SRL, E. Merck, Aldrich-sigma
and S.D. Fine.
2.3 Nitrate content measurement
The nitrate content of leaves was estimated using the
method given by Grover et al. (28). Half a gram of
fresh leaves was placed in a conical ask, to which 50
mg charcoal and 10 mL distilled water were added and
the ask was boiled for 45 minutes. After ltration,
the volume was made up to 50 mL by adding doubledistilled water; 1 mL of this aliquot was taken and
0.5 mL of 2 mM CuSO4 solution, 0.25 mL of 0.01 M
hydrazine sulfate and 0.25 mL of 0.1 N NaOH was
added. The vials were kept in a water bath incubator

for 10 minutes at 33C and then transferred onto ice

and 0.5 mL chilled acetone, 1.0 mL of 1% sulfanilamide and 0.02% NEDD [N(1-napthyl) ethylene
diamine dihydrochloride] were added. The volume was
made to 6 mL by adding 1.5 mL double-distilled water
and the vials were kept for 20 minutes for color
development. Absorbance was recorded at 540 nm on a
UVvis spectrophotometer (model DU 640 Beckman,
USA). The nitrate content was expressed as mmole/g
fr.wt. The concentration of nitrate was determined
against the standard curve prepared by using KNO3
(potassium nitrate) solution.
2.4 Nitrate reductase (EC 1.6.6.1) activity assay
Nitrate reductase (NR) activity was estimated using the
method given by Klepper et al. (29). Half a gram of
fresh leaves with 3.0 mL phosphate buffer (pH 7.2)
was kept in vials with 3.0 mL of 0.4 M KNO3. The
vials were kept in vacuum desiccators and vacuum
inltration was performed at 30-second intervals until
the leaves settled down. The vials were kept in a water
bath incubator for 1 hour at 33C under dark conditions. After incubation, the vials were kept in hot water
for 5 minutes to stop the reaction, then a 0.2-mL aliquot was taken and 1.0 mL of 1% sulfanilamide and
1.0 mL of 0.02% NEDD [N(1-napthyl) ethylene diamine dihydrochloride] were added. The nal volume
was made up to 6.0 mL by double-distilled water. The
vials were kept in dark for 20 minutes for color
development. Absorbance was recorded at 540 nm on a
UVvis spectrophotometer (model DU 640 Beckman,
USA). The corresponding concentration of nitrate was
determined against the standard curve of nitrate prepared by NaNO2 (sodium nitrite) solution. The NR
activity was expressed as l mole/g fr.wt./hour.
2.5 Soluble amino acids content determination
Lee and Takahashis method (30) was used for the
estimation of soluble amino acids. Half a gram of fresh
leaf material was ground in 5 mL of absolute ethanol
with the help of mortar and pestle and transferred to a
centrifuge tube. It was then centrifuged at 5000 rpm for
10 minutes at 4C. After that, the supernatant was
taken in a test tube and alcohol was evaporated by
incubating the tubes at 80C for 1 hour in a water bath.
Pellets were dissolved in 10 mL of 0.5 M citrate buffer
(pH 5.5) by vigorously using a vortex; 0.5 mL of aliquot was taken from this and 1.5 mL of 55% glycerol
and 0.5 mL of 1% ninhydrin solution were added. The
vials were kept in the water bath for 20 minutes at
100C and the development of a purple blue color was
observe. The volume in each vials were made up to
6 mL by adding 0.5 M citrate buffer (pH 5.5).

Journal of Essential Oil Research

Absorbance was recorded at 570 nm on a UVvis spectrophotometer (Model DU 640 B, Beckman, USA).
Calibration curve was prepared from glycine (Sigma)
of different concentrations to calculate the amino acid
content in different samples. The concentration was
expressed in mg/g fr.wt.

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2.6 Soluble protein content determination

The soluble protein content of different samples was
estimated following the method of Bradford (31); 0.5 g
of chopped fresh leaf material was homogenized in
1.5 mL of 0.1 M phosphate buffer, pH 7.0 (extraction
buffer) at/below 4C with the help of a precooled mortar and pestle, and then kept on ice during the process
of homogenization. The homogenate was transferred to
a 2-mL Eppendorf tube and centrifuged at 5000 rpm for
20 minutes at 4C. An equal amount of chilled 10%
TCA was added to 0.5 mL of the supernatant, which was
again centrifuged at 3300 rpm for 30 minutes. The
supernatant was discarded and the pellet left was washed
with acetone. It was then dissolved in 1 mL of 0.1 N
NaOH. To 0.2 mL aliquot, 1 mL of Bradfords reagent
was added and vortexed. The tubes were kept for 10
minutes for optimal color development. The absorbance
was then recorded at 595 nm on a Beckman spectrophotometer (DU 640, Fullerton, USA). The soluble protein
concentrations were quantied with the help of a standard curve prepared from the standard of bovine albumin
serum (BSA) from sigma, USA. The protein content was
expressed in mg/g fr.wt.
2.7 Elemental composition
Five plants were collected randomly from each treatment and were wiped free of any adhering dust using
double-distilled water. Different vegetative parts were
cut precisely into stem, leaves and roots and analyzed at
different developmental stages for assessing nitrogen (N)
and sulfur (S) composition. Each sample was dried for
48 hours in a hot air oven at 65C. The dried samples
were nely powdered and passed through a 72-mm
mesh screen. The powder was stored in polyethylene
bags, labeled and used for analysis later.
2.8 Estimation
Nitrogen (N) and sulfur (S) content (% dry weight)
were analyzed by packing the known weight of plant
sample-powder in tin boats with the help of an Elemantar system (CHNS Analyse, Vario EL.)
2.9 Oil Extraction and Analysis
The essential oil from 200 g P. corylifolia seeds was
isolated separately by hydrodistillation (2 hours) using

a Clevenger-type apparatus. The essential oils were dissolved in Et2O, dried over anhydrous MgSO4, ltered
and the solvent was removed by evaporation on a water
bath. The plant material was ltered off and the aqueous residues were extracted, rst with n-hexane
(250 mL) and then with Et2O (250 mL), to remove
traces of volatile compounds. The organic solvents, dissolved in the aqueous phase, were evaporated under
reduced pressure at room temperature. The remaining
aqueous extracts were treated with -glucosidase
(41 mg) from almonds (BioChemika, 1 g, Lot. 49290,
Fluka) at 28C for 16 hours and the volatile compounds were collected in n-hexane (0.5 mL) by
hydrodistillation. All the essential oils and the volatile
fractions were analyzed by GC and GCMS.
2.10 Gas chromatography
GC analyses were accomplished with a HP-5890 Series II
instrument equipped with HP-WAX and HP-DB-5
capillary columns (30 m 0.25 mm, 0.25 m lm thickness), working with the following temperature program:
60C for 10 minutes, rising at 5C/minute to 220C;
injector and detector temperatures, 250C; carrier gas,
nitrogen (2 mL/minute); detector, dual FID; split ratio,
1:30; injection, 0.5 mL. The identication of the components was performed, for both the columns, by the
comparison of their retention time with those of pure
authentic samples and by means of their linear retention
indices (Lri) relative to the series of n-hydrocarbons. The
relative proportions of the essential oil constituent percentages were obtained by FID peak-area normalization,
all relative response factors being taken as one.
2.11 Gas ChromatographyMass Spectrometry
GCMS analyses were performed with a Varian CP3800 gas chromatograph equipped with a DB-5 capillary column (30 m 0.25 mm; coating thickness 0.25
m) and a Varian Saturn 2000 ion trap mass detector.
Analytical conditions: injector and transfer line temperatures 220 and 240C, respectively; oven temperature programmed from 60 to 240C at 30C/minute;
carrier gas helium at 1 mL/minute; injection 0.2 L
(10% hexane solution); split ratio 1:30. Identication of
the constituents was based on comparison of the retention times with those of authentic samples, comparing
their linear retention indices relative to the series of
n-hydrocarbons, and on computer matching against
commercial indexes (NIST 98 and Adams) besides a
home-made library of mass spectra built up from pure
substances and components of known oils and MS literature data. Moreover, the molecular weights of all the
identied substances were conrmed by GCchemical
ionization MS (CIMS), using Me-OH as the CI gas.

S. Jan et al.

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2.12 Solid-phase particle microextraction (SPME)

analysis
SPME analysis was performed with a Supelco SPME
device, coated with polydimethylsiloxane (PDMS,
100 m), in order to sample the headspace of a portion
(1 g) of the seed sample. Each plant aliquot was
inserted into a 100-mL glass conical ask and allowed
to equilibrate for 20 minutes. After the equilibration
time, the ber was exposed to the headspace for 15
minutes at room temperature, and when the sampling
was nished (5 minutes), the ber was withdrawn into
the needle and transferred to the injection port of the
GC and GCMS system, operating in the same condition as described above for both the identication and
quantication of the constituents, apart from the splitless injection mode and the injector temperature
(250C).
2.13 Statistical analyses
The data are expressed as mean SE (n = 5). For differences between several mean values of the control
and exposed seedlings, analysis of variance (ANOVA)
was performed. Duncans multiple range test (DMRT)

test was used to determine whether the values were

signicantly different from the control.

3. Result and discussion

The data on biochemical and elemental parameters
reect substantial alteration in developing seedlings of
P. corylifolia L. induced by variable doses of gamma
rays. Presowing gamma irradiation at 2.5, 5 and
10 kGy when applied to dry seeds of P. corylifolia L.
signicantly increased the concentration of soluble protein, free amino acid and nitrate content, along with
NR activity, reaching maxima at owering stage (Tables
14). Soluble protein content varied from 32.12 mg/g
fr.wt. at the owering stage to 27.50 mg/g fr.wt. at
post-owering stages at 10 kGy. Higher gamma doses
had a depressing effect on protein content at all
developmental stages. Maximum decrease (41.88%)
was observed in plants raised from seeds irradiated
with 20 kGy at the pre-owering stage (Table 1). In
accordance with the results obtained by Stajner et al.
(32) in soybean seeds, 10-Gy dosage caused a slight
increase of about 11.0% in total soluble protein content
compared with the non-irradiated seeds. This study

Table 1. Variation in soluble protein content (mg/g fr.wt.) at various growth stages of Psoralea corylifolia L. exposed to different
doses of gamma rays.
Developmental stage
Treatments
Control
2.5 kGy
5 kGy
10 kGy
15 kGy
20 kGy

Pre-owering
18.67a
21.66
24.93
29.72
13.16
10.85

0.13
0.06
0.01
0.08
0.05
0.12

(16.01)b
(33.52)c
(59.18)d
(29.51)e
(41.88)f

Flowering
24.5g
25.7
26.7
32.12
28.5
29.1

0.01
0.01
0.01
0.03
0.01
0.01

(4.89)h
(8.97)i
(31.10)n
(16.32)k
(18.81)l

Post-owering
19.39m
23.51
26.66
27.5
14.56
11.70

0.23
0.09
0.02
0.01
0.08
0.03

(21.24)m
(37.49)mn
(12.24)j
(24.90)no
(39.65)o

Note: Values in parenthesis represent percent variation. Values with different superscripts are signicantly (p < 0.05) different from each other
(Duncans multiple range test). *p 0.05. The values represent mean SE (n = 5). CD at 5%. Treatments: 0.099*; developmental stages: 0.070*;
treatment developmental stages: 0.221*.

Table 2. Variation in free amino acid (mg/g fr.wt.) at various growth stages of Psoralea corylifolia L. exposed to different doses
of gamma rays.
Developmental stages
Treatments
Control
2.5 kGy
5 kGy
10 kGy
15 kGy
20 kGy

Pre-owering
12.71a
16.92
21.92
32.70
8.06ab
6.00bc

0.11
0.08
0.05
0.07
0.05
0.01

(33.12)a
(72.46)a
(157.27)a
(36.58)ab
(52.79)bc

Flowering
14.81c
18.49
24.54
38.76
11.76
8.87d

0.06
0.13
0.04
0.02
0.06
0.04

(24.84)c
(65.49)c
(161.38)cd
(20.59)d
(40.10)d

Post-owering
13.10d
17.41
14.55
11.55
7.67d
7.07d

30.60
28.10
25.56
21.06
14.13
21.65

(32.90)d
(11.06)d
(11.83)d
(41.45)d
(46.03)d

Note: Values in parenthesis represent percent variation. Values with different superscripts are signicantly (p < 0.05) different from each other
(Duncans multiple range test). *p 0.05. The values represent mean SE (n = 5). CD at 5%; treatments: 17.11*; developmental stages: 12.10*;
treatment developmental stages: 38.25*.

Journal of Essential Oil Research

Table 3. Variation in nitrate content (mmol/g fr.wt.) at various growth stages of Psoralea corylifolia L. exposed to different doses
of gamma rays.
Developmental stages
Treatments
Control
2.5 kGy
5 kGy
10 kGy
15 kGy
20 kGy

Pre-owering
4.62a
7.05
8.60
10.79
3.58
2.55

0.05
0.01
0.13
0.04
0.08
0.07

(52.59)b
(86.14)c
(133.54)c
(22.51)d
(44.80)e

Flowering
5.91f
8.21
10.67h
12.43
4.53
3.11

0.05
0.04
0.06
0.07
0.09
0.02

(38.91)g
(80.54)h
(110.32)i
(23.35)j
(47.37)k

Post-owering
5.54k
7.76l
9.59
11.55
3.80
2.76

0.13
0.06
0.12
0.06
0.03
0.05

(40.07)m
(73.10)m
(108.48)n
(31.40)o
(50.18)p

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Note: Values in parenthesis represents percent variation Values with different superscripts are signicantly (p < 0.05) different from each other
(Duncans multiple range test). *p 0.05. The values represent mean SE (n = 10). CD at 5%; treatments: 0.077*; developmental stages: 0.055*;
treatment developmental stages: 0.173*.

Table 4. Variation in nitrate reductase activity (mol/g fr.wt./hour) at various growth stages of Psoralea corylifolia L. exposed to
different doses of gamma rays.
Developmental stages
Treatments
Control
2.5 kGy
5 kGy
10 kGy
15 kGy
20 kGy

Pre-owering
31.28a
36.04
43.24
48.32
26.87
19.51

0.06
0.22
0.04
0.08
0.13
0.09

(15.21)b
(38.23)c
(54.47)c
(14.09)d
(37.62)e

Flowering
34.45f
39.26
48.16
52.62
31.66
23.82

0.06
0.05
0.12
0.05
0.18
0.03

(13.96)g
(39.79)h
(52.74)i
(8.09)j
(30.85)k

Post-owering
34.07
38.57
45.83
50.88
29.86p
22.56

0.02
0.23
0.04
0.03
0.07
0.13

(13.20)m
(34.51)n
(49.33)o
(12.35)p
(33.78)q

Note: Values in parenthesis represent percent variation. Values with different superscripts are signicantly (p < 0.05) different from each other
(Duncans multiple range test). *p 0.05. The values represent mean SE (n=10). CD at 5%; treatments: 0.159*; developmental stages: 0.112*;
treatment developmental stages: 0.355*.

Figure 1. Variation in total nitrogen (%) content per mg at various developmental stages of Psoralea corylifolia L. exposed to
different doses of gamma rays (T1=2.5, T2=5, T3=10, T4=15 and T5=20 kGy). Bars represents the mean SE (n = 10). Values
with different letters are signicantly (p < 0.05) different from each other (Duncans multiple range test).

further demonstrated that the introduction of carbonyl

groups into amino acids marked oxidative modication

of proteins in response to gamma ray exposure. This

general decline of protein synthesis prevents plastid

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S. Jan et al.

Figure 2. Variation in total sulfur (%) content per mg at various developmental stages of Psoralea corylifolia L. exposed to
different doses of gamma rays (T1=2.5, T2=5, T3=10, T4=15 and T5=20 kGy). Bars represents the mean SE (n = 10). Values
with different letters are signicantly (p < 0.05) different from each other (Duncans multiple range test).

transformation, resulting in greening and degreening

inhibition by irradiation (33).
The free amino acid content of the leaves showed a
signicant increase (p < 0.05) with age of the plant up
to owering with 10 kGy and varied response of free
amino acid pool with 20 kGy. Increased simple organic
molecules such as sugar, free amino acids and total soluble phenols might act as an osmotic for the regulation
of plant osmosis (6, 34). The observed increase in free
amino acid content due to low doses of gamma rays is
in agreement with ndings of Sattar et al. (35), who
documented increases in essential and non-essential
amino acids of soybean when irradiated at a dose level
of 0.10 kGy. Cowpea exhibited varied responses to
gamma exposure where a free amino acid pool
decreased signicantly with the exception of tyrosine
compared with their respective controls (36). Gamma
irradiation was found to be efcient in maintaining the
nutritional potential of Mucuna pruriens seeds (37).
The nitrate content and NR activity in the leaves
showed a signicant (p < 0.05) increase with plant age
in both control and irradiated plants. Maximum
enhancement (54.47%) in NR activity and (133.54%)
in nitrate content was recorded with 10 kGy at the
Table 5.

pre-owering stage. Nitrate content and NR activity in

leaves were higher at owering stage following low
doses of gamma rays raised to peak values at 10 kGy.
NR acivity in P. corylifolia L. decreases signicantly
with increase in gamma dosage applied, the maximum
reduction being observed at 20 kGy (Tables 4 and 5).
This observation is in agreement with results noticed in
Haworthia callus, Ustilago and Agrostemma gethago
by exposure to varied doses of gamma rays (6).
Nitrogen exhibited a maximum increase in roots
(101.02%), stems (67.28%) and leaves (64.83%),
recorded with 10 kGy at the owering stage, whereas
the maximum decline of stems (79.10%), leaves
(45.79%) and roots (44.07%) was recorded with
20 kGy (Figure 1). Our results are in agreement with
ndings of Maman et al. (38), in which the content of
nitrogen in most tissues of crop plants were high during the early stage of growth and decreased with
advancement of plant age. Deaf (14) reported an
increase in nitrogen, phosphorus and potassium content of lemongrass when the seeds were exposed to
14 krad gamma irradiation. Mahmoud (15) indicated
that gamma irradiation increased phosphorus and potassium content of Delphinium plants. Contradictory

Variation in essential oil content (%) in Psoralea corylifolia L. seeds following different gamma irradiation dosage.

Treatments

Control, mean SD

T1, 2.5 kGy

T2, 5 kGy

T3, 10 kGy

T4, 15 Gy

T5, 20 kGy

Essential oil

0.89 0.04

1.01 0.01
(13.48)d

1.1 0.06
(23.59)bc

1.3 0.38
(46.06)ab

1.49 0.29
(67.41)a

1.60 0.15
(79.77)a

Note: Each data represents the mean standard errors.Values with different superscripts are signicantly different from each other (ANOVA,
Tukeys test, p 0.05). Value in parenthesis indicates percent variation. *p 0.05. The values represent mean SE (n=10). CD at 5%; treatments:
essential oil, 0.129*, other volatile fraction, 0.113*.

Journal of Essential Oil Research

Table 6. GCMS analysis of the essential oil and other volatile fractions of the seeds of Psoralea corylifolia L. exposed to variable doses of gamma radiation.

Downloaded by [University of Kashmir], [Sumira Jan] at 22:10 01 April 2015

Essential oil (%)

a

Compounds

RIp

3-(E)-Hexen-1-ol
Heptanal
Tricyclene
-pinene
Camphene
-Pinene
Sabinene
1-Octen-ol
-Myrcene
Limonene
-Phellandrene
Linalool
Nonanal
Terpinen-4-ol
Geraniol
Geranial
Caryophyllene
cis-Piperitol
Thymol
Gernyl acetate
-Caryophyllene
-Gurjunene
-Humulene
Germacrene D
-Cadinene
Spathulenol
Caryophyllene oxide
Apiol
-Cadinol
Angelicin
Psoralen
Bakuchiol
(E)-Phytol
n-Docosane
Viridiorol
n-Heneicosane
Bicyclogermacrene
-Muurolol
allo-Aromadendrene
Humulene oxide-2
n-Bourbonene
-Terpineol
n-Hexadecane
trans--Cadinene
-(E)-Ocimene
Total (%)

845
903
924
936
949
987
976
986
984
1035
1036
1100
1107
1174
1227
1243
1428
1181
1274
1364
1417
1432
1454
1479
1532
1578
1586
1619
1643
1774
1829
1837
2099
2200
1593
2100
1496
1645
1463
1610
1386
1180
1600
1513
1038

Control

2.5 kGy

5 kGy

10 kGy

15 kGy

20 kGy

1363
1179
1010
1027
1087
1116
1130
1164
1453
1206
1208
1495
1394
1633
1787
1717
1593
1748
2162
1753
1593
1537
1709
1707
2150
2104
1962
2210
2102
2265
2297
2285
1027
2217
2211
2100
1767
2102
1683
2018
1530
1167
1608
1767
1253

0.69
0.48
9.8
52.2
3.6
0.58
0.43
0.8
4.2
0.6
0.30
1.9
0.4
0.9
0.6
0.9
6.5
0.8
0.4
0.6
18.5
0.3
4.6
6.8
0.9
0.9
1.9
0.78
0.68
6.24
24.8
22.78
0.9
0.4
0.5

0.86
0.89
10.2
54.12
5.01
0.69
0.5
0.90
5.10
0.90
0.43
2.5
1.4
0.9
0.79
1.4
7.5
1.2
0.8
0.9
21.0
0.9
5.0
7.0
1.4
1.2
2.3
0.82
0.72
6.58
25.0
23.80
1.5
0.6
1.2
1.3
1.0
0.6
0.9
0.8
2.0
0.9
0.7
1.0
1.3
209.59

0.70
0.50
9.9
53.7
4.8
0.69
0.49
0.90
4.7
0.90
0.37
2.4
0.7
1.5
0.77
1.6
7.6
1.2
0.7
0.9
19.5
0.8
5.0
7.01
1.5
1.5
2.4
0.78
0.75
7.08
25.96
25.93
1.5
1.5
1.6
1.5
1.2
0.7
0.9
0.8
2.2
1.2
0.8
0.9
1.6
211.93

0.90
0.94
11.3
56.12
6.09
0.76
0.90
0.95
5.15
0.95
0.46
2.80
1.2
1.9
0.89
1.56
8.09
1.90
0.90
1.2
22.09
10.7
5.32
8.56
2.34
1.9
2.8
0.98
0.80
7.90
26.04
24.09
1.90
0.9
1.9
1.65
1.7
0.8
1.2
0.85
2.4
1.3
0.9
1.2
1.7
239.62

0.94
0.98
12.34
58.90
7.98
0.79
0.94
1.45
6.34
1.45
0.56
3.06
1.5
2.09
0.97
1.89
8.98
2.34
1.23
1.87
23.19
11.87
7.02
9.06
4.34
2.07
2.78
1.45
0.98
7.76
27.04
25.09
2.03
1.0
2.5
1.7
1.9
0.97
1.7
0.95
2.65
1.87
1.45
1.67
1.98
266.2

1.23
1.45
14.54
60.90
8.98
0.89
1.04
1.05
7.74
1.87
0.87
4.98
1.98
2.87
1.34
2.09
9.04
2.89
1.67
1.98
25.87
14.89
7.08
9.009
4.64
2.17
2.89
1.78
1.09
7.89
37.04
35.09
2.53
1.56
2.67
1.85
2.09
1.23
1.67
1.23
2.89
1.90
1.69
1.87
2.01
308.96

RIp

0.7
0.2
0.6
0.2
1.9
0.4
0.3
0.1
1.3
185.16

results were also reported; for example, a decrease in

nitrogen content was found in Capsicum annum (39).
Zham and Voloozh (40), however, reported that
irradiating tomato seeds with 2.5 krad gamma rays did
not affect the concentration of nitrogen or phosphorus,
which is contradictory to our ndings.
The sulfur content in the roots, leaves and stems of
P. corylifolia L. also showed a signicant (p < 0.05)
enhancement with the age of the plant up to the ower-

ing stage, followed by pre-owering and post-owering

stages in control as well as irradiated plants. A signicant increase in sulfur content of leaves was recorded
with low doses of gamma rays. Maximum sulfur content was recorded in plants raised from seeds irradiated
at 10 kGy (Figure 2). Sulfur content was found to be
highest in leaves, stems and roots, respectively. At 15
and 20 kGy, there was marked decrease in sulfur
content, signifying sulfur to be most sensitive to

S. Jan et al.

Table 7. Gas chromatographymass spectrometry (GCMS) analysis after hydrolysis by -glucocidase in the seeds of Psoralea
corylifolia L.exposed to variable doses of gamma radiation.

Downloaded by [University of Kashmir], [Sumira Jan] at 22:10 01 April 2015

-glucosidase (%)
a

Control

2.5 kGy

5 kGy

10 kGy

15 kGy

20 kGy

4.6
11.2
27.1
31.4
1.1
8.1
5.4
6.4
26.5
3.2
2.1
0.76
0.5
3.8
1.5
133.66

5.98
12.01
28.01
30.03
1.9
8.9
6.0
7.0
28.02
3.8
4.5
1.8
0.90
4.68
1.69
146.22

6.4
12.7
28.5
30.5
1.70
9.1
7.8
6.8
28.6
4.5
4.3
0.90
0.94
4.56
1.63
148.90

7.09
13.09
30.09
31.45
1.98
10.03
7.9
7.09
29.05
4.76
4.90
1.90
1.87
4.98
1.76
157.94

8.05
14.29
31.19
32.05
2.03
11.78
8.03
7.23
30.08
5.09
5.60
2.0
1.97
5.18
1.86
166.43

9.07
15.09
32.17
33.08
2.23
12.08
9.13
8.23
31.88
6.19
6.60
2.23
2.24
5.35
1.95
177.52

Compounds

RIp

RIp

3-(E)-Hexen-1-ol
Tricyclene
-pinene
Camphene
1-Octen-ol
-Myrcene
Limonene
Caryophyllene
Germacrene D
-Cadinene
Spathulenol
n-Docosane
n-Eicosane
Bicyclogermacrene
n-Bourbonene
Total (%)

845
924
936
949
986
984
1035
1428
1479
1532
1578
2200
2100
1496
1386

1363
1010
1027
1087
1164
1453
1206
1593
1707
2150
2104
2217
2100
1767
1530

Note: RIpa refers to retention indices of compounds are listed in order of their elution from HP5-apolar column. RIpb refers to retention indices on
DB-WAX polar column.

gamma irradiation. Ionizing irradiation has a pronounced effect on protein sulfhydryl group as investigated in animals; however, there is a paucity of
research done on sulfur metabolism in plants (41). Cysteine, lysine and glutathione are known to increase after
low dose gamma irradiation (26, 42). Ionizing radiation
inhibited the utilization of methionine (43). Further,
Fan et al. (44) showed that gamma irradiation at doses
of pathogen inactivation signicantly increased the concentrations of most sulfur compounds, especially with
10 kGy. Ionizing radiation caused an increase in the
levels of phenylalanine ammonia lyase enzyme and
superoxide dismutase activity (26, 34). However, Jan
et al. (26) reported a decrease in glutathione with
increase in gamma irradiation doses.
GCMS and SPME analysis of volatile compounds
in P. corylifolia L. seeds exposed to variable doses of
gamma rays were summarized in Tables 68. A yellowish oil with a peppermint-like pleasant smell was
produced by hydro-distillation of seeds of P. corylifolia.
Oil content in Psoralea seeds showed statistically signicant differences among the treatments. Maximum oil
content (1.60%) was found in seeds irradiated with
20 kGy and minimum oil content was found in control
seeds (0.89%) as represented in Table 6. Our result
showed a high content of coumarins, phenylpropanoids
and isoavones, especially in seeds exposed to 15 and
20 kGy. Plants raised from seeds exposed to 20 kGy
exhibited the maximum increment in tricyclene
(48.36%), -pinene (16.66 %), camphene (149.44%),
-caryophyllene (39.83%), angelicin (26.44%), psoralen
(49.35%), bakuchiol (54.09%), gerianal (132.22%) and

germacrene D (33.67%). SPME analysis of seeds

conrmed tricyclene, -pinene, caryophyllene, germacrene D and camphene as the main constituents, and the
presence of aliphatic aldehyde, n-nonanal, an alcohol
(Z)-3-hexene-ol, and -(E)-ocimene, which were found
in traces in control seed essential oil. The percentage
of SPME analyses followed the same trend as that of
GCMS analyses. Plants raised from seeds exposed to
20 kGy exhibited maximum increment in tricyclene
(11.83%), -pinene (21.21%), camphene (51.40%),
caryophyllene (70.31%), -gurgenene (196.67%),
n-nonanal (31.81%), -(E)-ocimene (120.09%), (Z)3-hexene-ol (29.34%) and germacrene D (22.16%).
Tricyclene, - and -pinene, sabinene and germacrene D
constitute important components of the monoterpene
fraction. -Caryophyllene, angelicin, psoralen and bakuchiol compounds were the main components of the
volatile oil of Psoralea seeds according to the quantity.
Psoralen composition increased about twofold (49.35%)
with 15 and 20 kGy, as described in Tables 68. The
gamma irradiation doses of 15 and 20 kGy induced a
signicant increase in other volatile oils in comparison
with the control. The control sample had fewer levels of
all fty-four compounds, as conrmed by GCMS and
SPME analyses (Table 68). There were exceptional
changes, as observed in the volatile oil compounds contents, even at radiation doses of 5 and 10 kGy, conrmed safe toxicologically and nutritionally. These
ndings are in agreement with the results of Calucci
et al. (45) and Seo et al. (46). The most important
change could be observed at an ionizing dose of 20 kGy
(twice the recommended dose) resulting in a threefold

Journal of Essential Oil Research

Table 8. Variation in volatile fractions of seeds of Psoralea corylifolia L. plants developed from variably gamma irradiated seeds
using solid-phase particle microextraction (SPME) analysis.

Downloaded by [University of Kashmir], [Sumira Jan] at 22:10 01 April 2015

SPME analyses
Compounds

RIaa

RIpb

Control

2.5 kGy

5 kGy

10 kGy

15 kGy

20 kGy

3-(E)-Hexen-1-ol
Heptanal
Tricyclene
-pinene
Camphene
Sabinene
1-Octen-3-ol
-pinene
(Z)-3-Hexene-ol
Limonene
-Phellandrene
-(E)-Ocimene
Terpinolene
Linalool
n-Nonanal
Geraniol
Geranial
-Cubebene
-Vlangene
-Copaene
-Bourbonone
-Cubebene
Isocaryophyllene
-Gurajenene
Caryophyllene
-Cedrene
-Gurjenene
-(E)-Farnesene
-Humelene
alloaromadendrene
-Murrolene
Germacrene D
Bicyclogermacrene
-Muurolene
n-Pentadecane
-(E-E)-Farnesene
cis--Cadinene
trans--Cadinene
-Cadinene
Spathulenol
n-Hexadecane
Humulene oxide II
-Muurolol
-Cadinol
-Bisabolol
n-Heptadecane
n-Nonadecane
n-Heneicosane
Total (%)

854
906
928
942
952
978
985
1017
1036
1039
1040
1038
1084
1093
1109
1237
1258
1352
1375
1378
1386
1393
1406
1409
1429
1432
1437
1440
1454
1463

1363
1185
1009
1038
1082
1130
1457
1119
1316
1209
1219
1256
1287
1509
1398
1795
1729
1457
1487
1517
1530
1543
1529
1617
1608
1539
1669
1707
1681
1694

0.7
4.5
16.9
45.3
1.4
0.6

0.83
4.92
17.03
47.86
1.72
0.9

0.89
5.20
17.89
49.98
1.83
0.93

0.90
5.34
18.78
51.23
1.97
1.2

0.95
6.09
18.98
53.54
2.09
1.30

1.23
7.09
18.90
54.87
2.12
1.32

0.9
7.5
0.5
0.4
0.9

1.2
8.3
0.62
0.68
1.3

1.42
8.7
0.77
0.70
1.5

1.57
8.9
0.82
0.73
1.70

1.63
9.2
0.93
0.79
1.78

1.67
9.7
0.98
0.87
1.98

3.3

3.7

3.9

3.95

3.97

4.35

0.3
0.5

0.45
0.65

0.56
0.72

0.67
0.79

0.73
0.89

0.80
0.93

3.2

3.7

3.9

4.02

4.98

5.45

0.6

0.8

0.90

1.02

1.05

1.78

1478
1482
1497
1499
1502
1504
1512
1513
1524
1579
1603
1608
1645
1654
1687
1703
1900
2100

1719
1769
1729
1497
1735
1768
1767
2147
2149
2153
1602
2017
2104
2221
2043
1700
1900
2100

7.4
0.9

7.9
1.4

8.2
1.5

8.7
1.9

8.90
2.0

9.04
2.05

0.2
0.4
0.4
0.5

0.5
0.6
0.5
0.7

0.67
0.78
0.65
0.83

0.78
0.89
0.72
0.89

0.89
0.90
0.77
0.99

0.98
0.95
0.86
1.3

97.3

106.26

112.42

117.47

123.35

129.22

increase of caryophyllene, and a twofold increase in psoralen and bakuchiol in comparison with control (Table 8).
These changes can be explained through the irradiation
effects on terpenes. Irradiation does not show any thermal effect on the avor compounds but via oxidation or

hydroxylation of the aromatic ring in terpene, or via an

indirect effect, which can generate free radicals from the
water contained in seeds (47). The radicals generated
from irradiation can react with terpenes to produce terpene oxides and terpene alcohols. On the other hand,

Downloaded by [University of Kashmir], [Sumira Jan] at 22:10 01 April 2015

10

S. Jan et al.

terpenes, which are incorporated in most of the essential

oils, have the same skeleton structure but differ in their
functional groups, such as OH, CHO or COOH. Therefore, changes induced congurationally can occur by
high doses, including changes in the positions of the
double bonds and the functional groups to produce different compounds (48). There is evidence that phenol
contents of medicinal plants are increased when they are
irradiated with ultraviolet B (49), while gamma and electron beam ionizing radiation did not induce any detectable qualitative or quantitative signicant changes in the
contents and yields of essential oils immediately after
ionizing radiation of some medicinal herbs (50). Flavonoids in the methanol extract of medicinal plants relatively resist the effect of ionizing radiation compared
with other extracts but our results describe the radiosensitivity of isoavones like bakuchiol. This nding is
contrary to the early investigations in which avonoids
were reported unstable in aqueous solution (51). Moreover, Kozlowski et al. (52) reported that radiolysis of avonoids in ethanol or methanol solution resulted in the
formation of new antioxidants. Increments in oil content
can be enumerated as a useful commercial and medicinal
outcome due to various benecial effects of P. corylifolia L. seed oil. The benecial effects of its extract
have been explained by various authors. Arora et al. (53)
conrmed the radiomodulatory effects of P. corylifolia
L. seed extract owing to its higher electron donation
potential than ascorbic acid. They have further illustrated
the signicant protective efcacy observed in the range
2550 g/ml of said extract against radiation induced
lysis of human erthyrocytes. A twofold increment in psoralen is a major nding, as this compound has attracted
the interest of researchers due to the free radical potential
of P. corylifolia L. seeds (2, 54, 55).
Gyawali et al. (56) reported on a threshold dose at
15 kGy resulting in a 12.6329.33% decrease in the
total content of volatile compounds of dried onion but a
signicant increase was recorded in the 10-kGy
irradiated sample. They also reported that although the
content of the majority of volatile compounds of dried
Welsh onion was increased after different doses of
gamma irradiation (1, 3, 5, 10 and 20 kGy), the content
of major sulfur-containing compounds such as 1-propanethiol, 5-methyl thiazole, propenyl propyl disulde,
dipropyl disulde and 3,5-diethyl-1,2,4-trithiolane was
decreased after the process, which might be because of a
decrease in sulfur composition as represented in our
study (Figure 2). Sulfur-containing compounds such as
dimethyl disulde, dimethyl trisulde, methyl propyl
disulde and methyl propyl trisulde were highly
increased by irradiation at 10 kGy. Variyar et al. (57)
found that the essential oil constituents in nutmeg showed clear quantitative differences in -terpeniol,
1-terpinene-4-ol and myristicin, which increased, while

that of sabinene, -pinene and elimicin decreased in

comparison with the control sample. Less signicant
changes in the content of some of the essential oil
constituents between the control and irradiated samples
were noted in clove and cardamom (57).
Mexis and Kantominas (58) reported a progressive increase in 2-pentanone, 1-hexanol, hexanal, and
nonanal contents of hazelnuts with irradiation doses.
Yalcin et al. (59) reported decrease in styrene and
p-xylene contents, whereas 1-hexanol content
decreased only at 7 kGy. Similarly, Fan and Gates
(44) found that irradiation of orange juice reduced
the concentration of acyclic monoterpenes such as
geranial, neral, myrcene, linalool after one and seven
days at irradiation dose of 2.24 kGy. The concentrations of monocyclic monoterpenes such as -pinene,
were not inuenced by irradiation, whereas valencene, a bicyclic sesquiterpene, was also found to be
resistant to irradiation. In contrast to our study, signicant quantitative changes were noted by Variyar
et al. (57) in some of the phenolic acids in clove
and nutmeg upon irradiation with a 10-kGy dose.
They reported that the content of gallic and syringic
acids in irradiated clove was increased, whereas that
of -coumaric, ferulic and synapic acids decreased to
approximately half to that of control spice and that
of caffeic and gentisic acids remained unchanged.

4. Conclusion
Overall, the present investigation concluded that presowing gamma irradiation of Psoralea corylifolia L.
could substantially improve the biochemical aspects
and elemental composition up to 10 kGy. However,
higher doses of gamma irradiation might lead to a
deterioration in biochemical and elemental aspects,
especially sulfur content, which seems to be the most
sensitive to gamma irradiation. Low as well as high
doses of gamma irradiation were effective in increasing
the variability for essential oil content. The variability
was obliquely directed towards high levels of essential
oil components, thus improving the oxidative stability
of P. corylifolia L. The yield and quality of oil at 15
and 20 kGy were enhanced signicantly, indicating that
the genes responsible for oil yield and quality get
affected and produce micromutations. Elicitation of
secondary metabolites has application in the overproduction of desired compounds, which is an area of
commercial importance especially for high-value and
low-volume products.
Acknowledgements
I will like to thank faculty members of INMAS for providing
me the Gamma Cell facility required for irradiation of seeds

Journal of Essential Oil Research

and Ajay Kumar, Incharge AIRF facility, JNU, for GCMS
and SPME analysis. All benets in any form from a commercial party related directly or indirectly to the subject of this
manuscript or any of the authors has been acknowledged. The
authors declare that they have no conict of interest.

Downloaded by [University of Kashmir], [Sumira Jan] at 22:10 01 April 2015

References
1. P.S. Phirke, A.B. Kudbe and S.P. Umbarkar, The
inuence of magnetic eld on plant growth. Seed Sci.
Technol., 24, 375392 (1996).
2. S. Jan, T. Parween, T.O. Siddiqi and Mahmooduzzafar,
Gamma irradiation induced qualitative and quantitative
changes in growth and productivity with special reference
to enhanced production of psoralen. Plant Growth
Regul., 64, 163171 (2011).
3. S.O. Fapohunda, S.T. Anjorin, E.C. Akueche and B.
Harcourt, Multi mycotoxin prole of gamma-radiated
sesame seeds from Abuja markets. Nigeria using LC-MS/
MS. Nat. Sci., 10, 127134 (2012).
4. A. Anisimov, V. Vorobev and A. Zuikov, The inuence
of laser radiation on the velocity of rotational motion of
protoplasm in Elodea cells. Laser Phys., 7, 11321137
(1997).
5. J. Podleny, Studies on inuence of laser light on seeds,
growth, development and yielding of the white lupine
(Lupinus albus L.) plants (in Polish). Monograe
Rozprawy Naukowe, IUNG, Puawy, 3, 159 (2002).
6. R.H. Moussa, Gamma irradiation regulation of nitrate
level in rocket (Eruca vesicaria subsp. sativa) plants. J.
New. Seed., 8, 91101 (2006).
7. S. Jan, T. Parween, T.O. Siddiqi and Mahmooduzzafar,
Effect of gamma radiation on morphological, biochemical
and physiological aspects of plants and plant products.
Environ. Rev., 20, 1739 (2012).
8. H.R. Moussa, Low dose of gamma irradiation enhanced
drought tolerance in soybean. Bulg. J. Agric. Sci., 17,
6372 (2011).
9. A.M.R. Afy, H.S. El-Beltagi, A.A. Aly and A.E.
El-Ansary, The impact of -irradiation, essential oils and
iodine on biochemical components and metabolism of
potato tubers during storage. Not. Bot. Horti Agrobo.,
40, 129139 (2012).
10. A.P. Casarett, Effects of radiation on higher plants and
plant communities. Ann. NY Acad. Sci., 59, 514 (1968).
11. L.V. Selenia and O.G. Stepanenko, Effect of pre-sowing
gamma irradiation on the productivity and active principle content of Matricaria recutita. Rastitelnye Ressusy,
15, 143154 (1979).
12. V. Zheljazkov, A. Margine, T. Stovea and K. Shetty,
Effect of gamma irradiation on some quantitative
characteristics in mint and cornmint. Acta Hort., 426,
381388 (1996).
13. A.A. Youssef and A.Z. Moussa, Effect of gamma rays on
growth and essential oil composition of chamomile
(Chamomilla recutita L.). Arab. Univ. J. Agric. Sci., 6,
301311 (1998).
14. N.S.D. Deaf, Chemical and biological studies on some
wild plants. M.Sc. Thesis, Fac. Agric., Zagazig Univ.,
Zagazig, Egypt (2000).
15. F.A.N. Mahmoud, Effect of gamma radiation and some
agrochemicals on germination, growth and owering of
Delphinium ajacis and Mathiola incana plants. M.Sc.
Thesis. Fac. Agric., Moshtohor, Zagazig Univ., Egypt
(2002).

11

16. J.M. Krenisky, J. Luo, M.J. Reed and J.R. Carney,

Isolation and antihyperglycemic activity of bakuchiol
from Otholobium pubescens (Fabaceae) a Peruvian
medicinal plants used for the treatment of diabetes. Biol.
Pharm. Bull., 22, 11371140 (1999).
17. P. Zhou, Y. Takaishi, H. Duan, B. Chen, G. Honda, M.
Itoh, Y. Takeda, O.K. Kodzhimatov and K.H. Lee, Coumarins and bicoumarin from Ferula sumbul: Anti-HIV
activity and inhibition of cytokine release. Phytochemistry, 53, 689697 (2000).
18. Y. Shikishima, Y. Takaishi, G. Honda, M. Ito, Y. Takeda,
O.K. Kodzhimatov, O. Ashurmetov and K.H. Lee,
Chemical constituents of Prangos Tshimganica; structure
elucidation and absolute conguration of coumarin and
furanocoumarins derivatives with anti-HIV activity.
Chem. Pharm. Bull., 49, 877880 (2001).
19. K. Bapat, G.J. Chintalwar, U. Pandey, V.S. Thakur, H.D.
Sarma and G. Samuel, Preparation and in vitro
evaluation of radioiodinated bakuchiol as an antitumour
agent. Appl. Rad. Isotopes, 62, 389393 (2005).
20. H. Haraguchi, J. Inonui, Y. Tamura and K. Mizutani,
Antioxidative components of Psoralea corylifolia L.
(Leguminosae). Phtytochem. Res., 16, 535544 (2002).
21. N.A. Khatune, M.E. Islam, M.E. Haque, P. Khoudhkar
and M.M. Rahman, Antibacterial compounds from seeds
of Psoralea corylifolia L. Fitoterapia, 75, 228300 (2004).
22. G. Jiangninga, W. Xinchub, W. Houb, L. Qinghuab and
B. Kaishun, Antioxidants from Chinese medicinal herb
Psoralea corylifolia. Food Chem., 91, 287292 (2005).
23. B. Manohar and K. Udaya Sankar, Prediction of solubility of Psoralea corylifolia L. seed extract in supercritical
carbon dioxide by equation of state models. Theor.
Found. Chem. Eng., 45, 409419 (2011).
24. M. Faisal and M. Anis, Thidiazuron induced high
frequency axillary shoot multiplication in Psoralea
corylifolia. Biol. Plant., 50, 437440 (2006).
25. M. Anis and M. Faisal, In vitro regeneration and mass
multiplication of Psoralea corylifolia An endangered
medicinal plant. Indian J. Biotechnol., 4, 261264 (2005).
26. S. Jan, T. Parween, T.O. Siddiqi and Mahmooduzzafar,
Antioxidant modulation in response to gamma induced
oxidative stress in developing seedlings of Psoralea
corylifolia L. J. Environ. Radioact., 113, 142149 (2012).
27. H. Fricke and E. Hart, Chemical dosimetry. In: Radiation
Dosimetry, vol. 2. Edits., F.H. Attix and. W.C. Roesch,
Academic Press, New York, NY (1996).
28. H.I. Grover, T.V.R. Nair and Y.B. Abrol, Nitrogen metabolism of upper three leaf blades of wheat at different soil
nitrogen levels, NR activity and contents of various nitrogenous constituents. Plant Physiol., 42, 287292 (1978).
29. L. Klepper, D. Flesher and R.H. Hageman, Generation of
reduced nicotinamide adenine dinucleotide for nitrate
reduction in green leaves. Plant Physiol., 48, 580590
(1971).
30. Y.P. Lee and T. Takahashi, An improved colorimetric
determination of amino acid with the use of ninhydrin.
Anal. Biochem., 14, 7177 (1966).
31. M.M. Bradford, A rapid and sensitive method for the
quantication of microgram quantities of protein utilizing
the principle of protein-dye-binding. Analyt. Biochem.,
72, 248259 (1976).
32. D. Stajner, M. Milosevic and B.M. Popovic, Irradiation
effects on phenolic content, lipid and protein oxidation
and scavenger ability of soybean seeds. Int. J. Molec.
Sci., 8, 618627 (2007).

Downloaded by [University of Kashmir], [Sumira Jan] at 22:10 01 April 2015

12

S. Jan et al.

33. E. Kovacs and A. Keresztes, Effect of gamma and UV-B/

C radiation on plant cells. Micron, 33, 199210 (2002).
34. S. Jan, T. Parween, T.O. Siddiqi and Mahmooduzzafar,
Enhancement in furanocoumarin content and phenylalanine ammonia lyase activity in developing seedlings of
Psoralea corylifolia L. in response to gamma irradiation
of seeds. Rad. Environ. Biophys., 51, 341347 (2012).
35. A. Sattar, X. Neelofar and M.A. Akhtar, Effect of radiation and soaking on phytate content of soybean. Acta
Aliment., 19, 331336 (1990).
36. O.A. Joseph, M. Klasus, D. Kwaku and M. Amanda,
Functional properties of cowpea (Vigna unguiculata L.
Walp) ours and pastes as affected by gamma
irradiation. Food Chem., 93, 103111 (2005).
37. R. Bhat, K.R. Sridhar and S. Seena, Nutritional quality
evaluation of velvet bean seeds (Mucuna pruriens) exposed to gamma irradiation. Int. J. Food Sci. Nutr., 59,
261278 (2008).
38. N. Maman, S.C. Mason, T. Galusha and M.D. Clegg,
Hybrid and nitrogen inuence on pearl millet production
in Nebraska: Yield, growth, and nitrogen uptake, and
nitrogen use efciency. Agro. J., 91, 737743 (1999).
39. J. Iqbal, M. Kutacek and V. Jiracek, Effects of acute
gamma irradiation on the concentration of amino acids
and protein-nitrogen in Zea mays. Rad. Bot., 14,
165172 (1974).
40. D. Zham and D. Voloozh, The effect of pre-sowing
gamma irradiation of seeds on the yield and productivity
of outdoor tomatoes. Biologia, 31, 914 (1976).
41. C. Jo and D.U. Ahn, Production of volatile compounds
from irradiated oil emulsion containing amino acids or
proteins. J. Food Sci., 65, 612616 (2000).
42. S. Khare, A. Trivedi, P.C. Kesavan and R. Prasad, Effect
of gamma-radiation on the structure and function of
yeast membrane. Int. J. Rad. Bio., 42, 369383 (1982).
43. M. Horvatic and M. Gruner, The effect of gamma radiation on the methionine and tryptophan content of soy
protein products. Nahrung, 37, 14752 (1993).
44. X. Fan and R.A. Gates, Degradation of monoterpenes in
orange juice by gamma radiation. J. Agric. Food Chem.,
49, 24222426 (2001).
45. L. Calucci, C. Pinzino, M. Zandmeneghi, A. Capocchi,
S. Ghiringhelti, S. Tozzi and L. Galleschi, Effect of
-irradiation on the free radical and antioxidant contents
in nine aromatic herbs and spices. J. Agric. Food Chem.,
51, 927934 (2003).
46. H. Seo, J. Kim, H. Song, D. Kim, M. Byun, J. Kwon
and K. Kim, Effects of -irradiation on the yields of
volatile extracts of Angelica gigas Nakai. Rad. Phys.
Chem., 76, 18691874 (2007).
47. W.M. Urbain, Food Irradiation. Academic Press, London
(1986), p. 351.

48. J. Sadecka, Inuence of two sterilisation ways, gammairradiation and heat treatment, on the volatiles of black
pepper (Piper nigrum L.). Czech J. Food Sci., 28, 4452
(2010).
49. R. Kumari, S. Singh and S.B. Agrawal, Responses of
ultraviolet-B induced antioxidant defense system in a
medicinal plant, Acorus calamus L. J. Environ. Biol., 31,
907911 (2010).
50. M. Haddad, M.F. Herent, B. Tilquin and J.
Quetin-Leclercq, Effect of gamma and e-beam radiation
on the essential oils of Thymus vulgaris thymoliferum,
Eucalyptus radiata and Lavandula angustifolia. J. Agric.
Food Chem., 55, 60826086 (2007).
51. S. Scalia and M. Mezzena, Incorporation of quercetin in
lipid microparticles: Effect on photo and chemical-stability. J. Pharmaceut. Biomed. Anal., 10, 9094 (2009).
52. D. Kozlowski, P. Marsal, M. Steel, R. Mokrini, J.L.
Duroux, R. Lazzaroni and P. Trouillas, Theoretical
investigation of the formation of a new series of
antioxidant depsides from the radiolysis of avonoid
compounds. Rad. Res., 168, 243252 (2007).
53. R. Arora, A.S. Dhakerb, M. Adhikarib, J. Sharma, R.
Chawla, D. Gupta, A. Zhelevaa, Y. Karamalakovaa, R.
Kumarb, R.J. Sharmac, A. Sharmab, S. Sultana, R.P.
Tripathi, R.K. Sharma and V. Gadjevaa, Radical scavenging and radiomodulatory effects of Psoralea corylifolia
Linn. substantiated by in vitro assays and EPR
spectroscopy. Z. Naturfor. C. J. Biosci., 66, 35 (2011).
54. S. Adhakari, K.I. Priyadarsini and T. Mukherjee, Physico-chemical studies on the evaluation of the antioxidant
activity of herbal extracts and active principles of some
Indian medicinal plants. J. Clin. Biochem. Nutri., 40,
174183 (2007).
55. S. Nirmala, S. Anurag, M. Yadav, Psoralea corylifolia L.,
an endangered medicinal plant with broad spectrum
properties. Med. Plants Int. J. Phytomed. Rel. Indust., 6,
1320 (2014).
56. R. Gyawali, H. Seo, H. Lee, H. Song, D. Kim, M. Byun
and K. Kim, Effect of -irradiation on volatile
compounds of dried welsh onion (Allium stulosum L.).
Rad. Phys. Chem., 75, 322328 (2006).
57. P. Variyar, C. Bandyopadhyay and P. Thomas, Effect of
gamma irradiation on the phenolic acids of some Indian
spices. Int. J. Food Sci. Technol., 33, 533537 (1998).
58. S.F. Mexis and M.G. Kontominas, Effect of gammairradiation on the physiochemical and sensory properties
of hazelnuts (Corylus avellana L.). Rad. Phys. Chem., 78,
407413 (2009).
59. H. Yalcin, I. Ozturk, M. Hayta, O. Sagdic and T. Gumus,
Effect of gamma irradiation on some chemical
characteristics and volatiles content of linseed. J. Med.
Food, (2011). doi:10.1089/jmf,2010.0249.