Journal Journal

of Applied
Appl Journal of Applied Horticulture, 25(3): 319-324, 2023
Horticulture
https://doi.org/10.37855/jah.2023.v25i03.57 ISSN: 0972-1045

Anti-stress effects of melatonin pre-treatment on germination

indicators in tomato (Lycopersicon esculentum L.) under
polyethylene glycol induced drought stress

K.A. Mumithra Kamatchi1, A. Senthil1, M.K. Kalarani2, R. Swarnapriya3, S. Marimuthu4 and

M. Djanaguiraman1*
1
Department of Crop Physiology, TNAU, Coimbatore – 641003, India. 2Directorate of Crop Management, Tamil Nadu
Agricultural University, Coimbatore – 641003, India. 3Floriculture Research Station, Thovalai – 629302, India. 4Department
of Agronomy, AC & RI, Eachangkottai, Thanjavur – 631006, India. *E-mail: [email protected]

Abstract
Tomato is a commercially grown vegetable crop that is susceptible to drought stress, which can be improved by understanding the
tolerance mechanism. This study aims to evaluate the impact of melatonin pretreatment on alleviating PEG-induced drought stress on
tomato. Initially, screening was performed with different polyethylene glycol (PEG) concentrations (-0.1, -0.2, -0.3, and -0.4 MPa),
along with a control. The seeds treated with -0.3 MPa PEG exhibited lower germination percentage, seedling growth, and vigour index,
while no germination was observed at -0.4 MPa. Therefore, -0.3 Mpa was selected as the sublethal osmotic concentration to assess
the effect of melatonin to alleviate the osmotic stress. The seeds treated with various melatonin concentrations (20, 40, 60, 80, 100,
and 120 µM) and untreated stress control were subjected to -0.3 MPa osmotic concentration, and absolute control was maintained.
Seed germination parameters viz., germination indicators, shoot and root length indicators, biomass indicators, stress indices, and
germination speed, were recorded. The results showed that the germination parameters were significantly reduced in stress control;
however, notable improvements were observed in melatonin pre-treated seeds. Pretreatment with 100 µM melatonin led to an increase
in germination percentage (61%), germination index (150.5), vigour index (1010.1), promptness index (65), shoot length (6.28 cm),
root length (7.38 cm), fresh weight (0.66 mg), dry weight (0.07 mg), shoot length stress index (86), root length stress index (112), dry
matter stress index (87), rate of germination (1.5% per day), germination rate index (15.1% per day), and coefÏcient of the velocity of
germination (20.6) under drought stress. The study evidenced that melatonin pretreatment could effectively enhance seed germination
under PEG-induced drought stress in tomato, which, therefore, can be recommended for further research.
Key words: germination parameters, speed of germination, polyethylene glycol, melatonin, tomato

Introduction global warming, it is imperative to enhance seed germination

and seedling growth under low moisture conditions (Tripathy
Tomato (Lycopersicon esculentum L.) belongs to the Solanaceae et al., 2023).
family, and is widely recognized as one of the most valuable and
economically grown vegetable crops worldwide. It is widely The successful establishment of crops is contingent upon the
consumed as a staple food because of its health benefits and timing and duration of seed germination, particularly in low water
potential to treat various chronic diseases (Marti et al., 2016). conditions. Tomato seeds typically germinate within five days,
Tomato is the second largest vegetable crop, covering 4.85 mha while water stress can impede or delay germination due to various
biochemical changes (Mohammadizad et al., 2013). In this
after potato. Asia is leading in tomato production, accounting
perspective, drought stress can be simulated using polyethylene
for 61.1% of the global output, surpassing the Middle East,
glycol (PEG), and the application of plant hormones has proven
Africa, the United States and Brazil (Faostat, 2019). In India,
to be an effective approach for regulating seed germination
tomato cultivation spans an area of 8.41 lakh ha, with an annual
(Mahpara et al., 2022). Melatonin, an amphiphilic indoleamine
production of 20 million metric tonnes (Indiastat, 2022). Climate compound, is known for its environment-friendly properties and
change poses significant challenges to crop productivity, with role in enhancing plant tolerance to abiotic stress. In addition to its
abiotic stress emerging as a major constraint that adversely affects neurohormonal functions, melatonin plays a multifaceted role in
crop yield by 50% or higher. The 21st century has witnessed seed germination (Awan et al., 2023), vegetative and reproductive
increased climatic variations, leading to more frequent drought development (Wang et al., 2022). Several studies have shown that
occurrences, which are projected to rise by 3 to 8% (Tripathy et treating seeds with melatonin promotes germination and seedling
al., 2023). Mainly, vegetable crops are susceptible to drought growth in various crops, including wheat (Guo et al., 2022), rice
stress from germination to maturity. InsufÏcient soil moisture (Megala et al., 2022), green gram (Anitha et al., 2022) and cotton
during sowing significantly hampers the germination process, (Bai et al., 2020). Moreover, previous research has demonstrated
leading to poor growth and development, ultimately reducing that exogenous melatonin application can alleviate oxidative
yield (Toscano et al., 2023). To address the challenges posed by stress under drought conditions (Liu et al., 2015). However,
Journal of Applied Horticulture (www.horticultureresearch.net)
320 Anti-stress effects of melatonin pre-treatment on germination indicators in tomato

understanding the physiological and biochemical mechanisms (20, 40, 60, 80, 100 and 120 µM) for six h. To determine optimum
underlying drought tolerance in seed germination is essential, melatonin concentration to alleviate the PEG-induced drought
particularly for vegetable crops like tomato. stress, 25 seeds were placed on a Petriplate containing filter paper,
to which 15 mL of -0.3 MPa PEG 6000 (sublethal concentration)
The changes observed in germination indicators depend on the
was added. Each treatment was replicated four times. Eight
concentration of melatonin and the intensity of drought stress.
treatments were used in this experiment: absolute control (AC)
Nevertheless, pre-treating with melatonin can contribute to
with distilled water (without PEG), stress control (SC) with -0.3
regulating biochemical and metabolic reactions involved in
MPa of PEG and the different concentrations (20, 40, 60, 80, 100
the germination process (Zhang et al., 2020). Several studies and 120 µM) of melatonin (Mel) treated seeds exposed to osmotic
have documented the positive effect of melatonin in mitigating potential of -0.3 MPa. The germination indicators, shoot and root
drought stress (Annadurai et al., 2023). However, little is known growth indicators, biomass indicators, stress indices, and the
regarding the potential effects of melatonin concentrations speed of germination were recorded on 12th day. The germination
on germination indicators in tomato. Consequently, our study percentage was recorded on 2nd, 4th, 6th, 8th, 10th and 12th day.
aimed to examine the effects of melatonin pretreatment in the
tomato variety ‘PKM1’, focusing on achieving consistent seed Calculation of germination parameters:
germination, which is crucial under drought stress conditions. Germination indicators: Germination percentage (GP) indicates
The objectives of our investigation were twofold: (i) determining the proportion of seeds germinating on the 2nd, 4th, 6th, 8th, 10th and
the lethal and sub-lethal doses of PEG for seed germination, (ii) 12th day out of the total number of seeds sown in Petriplates. The
identifying the optimal concentration of melatonin pretreatment calculation of the germination percentage followed the method
for characterizing the seed germination, and seedling growth outlined in Nezar and Aldahadha, (2022).
indicators. This study sets the dimension on how melatonin pre-
treatment regulates seed germination, elucidating the underlying
physiological mechanisms. Moreover, our research could explore
the significance of melatonin seed treatment in aiding seedlings Vigour index (VI) was calculated according to the method
to overcome the negative effects of abiotic stress. mentioned in Hussain et al. (2015).
Vigour Index = Germination percent x Seedling length
Materials and methods
Germination index (GI) was calculated as per the method
Test Material: The tomato seeds used in the experiment were described in Hussain et al. (2015)
‘PKM1’ variety, provided by the Department of Vegetable
Science, Horticulture College and Research Institute, Coimbatore. GI = (12 x nl) + (11 x n2) 4- (10 x n3) ••• (1 x nl2)
Melatonin was obtained from Sigma-Aldrich Pvt. Ltd. India, where n1, n2, ..., n10 denote the number of seeds that germinated
stored at a temperature of -20 °C, and used when required. on the first, second, and subsequent days up to the twelfth day.
Experiment and treatment details Promptness index (PI) calculates the percentage of seeds that
germinate on the 2nd, 4th, 6th, 8th, 10th, and 12th day of observation,
Experiment I (Screening for optimum PEG concentration):
represented by nd2, nd4, nd6, nd8, nd10 and nd12, respectively,
The experiment was conducted during January 2022 in the as described in Raza et al. (2012).
Laboratory of the Department of Crop Physiology at Tamil Nadu
PI=nd2(1.20)+nd4 (1.0)+nd6(0.8)+nd8(0.6)+nd10(0.4)+nd12(0.2)
Agricultural University, Coimbatore, India. The tomato seeds
were surface sterilized with 5% sodium hypochlorite solution for Germination stress index (GSI) was determined using the
5 min, then washed three to four times with distilled water. The following formula:
following treatments were used: Absolute Control with distilled
water (without PEG) and four levels of PEG concentrations used
to impose stress at -0.1, -0.2, -0.3 and -0.4 MPa, respectively. Each Shoot and root growth indicators: The average shoot length
treatment had five replicates maintaining 25 seeds per replicate (SL) and root length (RL) were determined by measuring 25
and the sterilized seeds were placed in a petriplate containing seedlings from each replication within a treatment. The shoot
filter paper, to which 15 mL of different PEG solutions were length was measured from the base of the root to the tip of the
added. To evaluate the sublethal dosage of polyethylene glycol, shoot, while the root length was measured based on the length
tomato seeds with 2 mm of radical were considered germinated of the main root and expressed in cm.
(Florido et al., 2018). On the 12th day, the germination parameters Biomass indicators: Immediately after analyzing the growth
viz., rate of germination, shoot length (SL), root length (RL), and indicators, the fresh weight (FW) of the seedlings was recorded.
vigour index (VI), were recorded. The germination percentage Subsequently, the seedlings were subjected to a hot air oven at
was recorded on 2nd, 4th, 6th, 8th, 10th and 12th day. 60 °C for 48 h and the dry weight (DW) was recorded.
Experiment II (Screening for optimum melatonin Stress indices: The stress indices were calculated following the
concentration under PEG-induced drought): The melatonin methodology outlined in Raza et al. (2012). The shoot length
stock solution is prepared by dissolving the required quantity of stress index (SLSI) and root length stress index (RLSI) were
melatonin in 99.9% ethanol and made to a final volume of 1L with calculated from the measured shoot, root length of stress and
distilled water. The dilutions of different melatonin concentrations control seedlings.
were prepared from the stock solution. The sterilized seeds were
soaked in a beaker containing different melatonin concentrations
Journal of Applied Horticulture (www.horticultureresearch.net)
 Anti-stress effects of melatonin pre-treatment on germination indicators in tomato 321

The dry matter stress index (DMSI) was calculated using the Table 1. Effect of PEG-6000 on rate of germination, seedling growth
measured dry weight obtained after drying the samples in a hot (SL+RL) and vigor index (VI) in tomato
air oven at 60 °C. The calculation of DMSI was based on the dry Treatments Rate of Shoot length Root length Vigor index
weight between the stress and control seedlings. germination (cm) (cm)
Absolute 1.88 ± 8.54 ± 7.37 ± 1438.10 ±
control 0.020a 0.05a 0.096a 20.52a
Speed of germination: Germination rate index (GRI) is -0.1 MPa 1.57 ± 7.22 ± 7.76 ± 1126.24 ±
0.031b 0.09b 0.092a 21.97b
determined by calculating the percentage of germination on the
-0.2 MPa 1.30 ± 5.22 ± 5.98 ± 698.48 ±
2nd, 4th, 6th, 8th, 10th and 12th day, represented by G2, G4, G6, 0.020c 0.09c 0.106b 10.05c
G8, G10 and G12, respectively using the following formula as
mentioned in Rupal et al. (2020). -0.3 MPa 0.83 ± 3.28 ± 3.94 ± 289.50 ±
0.045d 0.08d 0.107c 18.44d
*No germination was observed in -0.4 MPa. The data represent the mean
of replications and standard error of means (SEM). The least significant
Rate of germination was calculated by counting the number of difference test (LSD5%) was computed to compare the means among
germinated seeds on the 12th day of the germination process and different treatments. The significance level was denoted using small
expressed in days (Bakhshandeh et al., 2017). letters, given that the means with the same letters are not statistically

120
Absolute cont rol -0.1 MP a
CoefÏcient of velocity of germination (CVG) is a measure that 100 -0.2 MPa -0.3 MP a
determines the frequency at which the seeds germinate on each
day (N), and the time took for seed N to germinate (T), starting Germination percentage (%) 80
from the day of sowing. It is calculated using the formula
established by Talska et al. (2020). 60

40

Data Analysis: The study was carried out in a completely 20

randomized block design (CRD). The germination parameters
were analyzed using analysis of variance (ANOVA) using SPSS 0
for Windows, version 16.0, developed by SPSS Inc., Chicago, 2 4 6 8 10 12

USA. The data is presented as the mean of replications and Germinati on time (days)

standard error of means (SEM). The least significant difference Fig. 1. Effect of PEG-6000 on germination percentage in tomato on 12th
test (LSD5%) was used to compare the means among different the treatments, -0.3 MPa (PEG) imposed seeds treated with
treatments. The significance level was denoted using small letters,
100 µM, 120 µM and 80 µM melatonin showed significant (P
given that the means with the same letters are not statistically
< 0.05) increase in germination percentage by 61%, 57% and
significant at p = 0.05. High-resolution graphs representing the
observed variables and the correlation among the germination 54% respectively than stress control. Notably, an increased
parameters were assessed using Pearson correlation in GraphPad concentration of melatonin (120 µM) exhibited an increased
Prism software for Windows, version 9.0.0. GP and showed a decreasing trend of GP. A direct correlation
supports this data; under decreased water potential, when
Results
melatonin concentration increases, the GP increases with the
Effect of PEG on germination parameters: The germination same intensity (Fig. 5). In the present study, it is observed that GP
percentage (GP), shoot length (SL), root length (RL), vigor index showed a significant positive relationship with MGT (r2=1.00),
(VI) and rate of germination was calculated on the 12th day and
PI (r2=0.99), GRI (r2=0.99), GI(r2=0.98) and VI (r2=0.98).
had a significant difference (P < 0.05) among the treatments. The
increased GP was observed in absolute control (90%), attaining its 140 AC SC
final germination on the 7th day. However, higher concentrations 20 µM Mel 40 µM Mel
of PEG -0.3 MPa showed decreased GP (40%) with a delay in 120 60 µM Mel 80 µM Mel
Germination percentage (%)

germination time, attaining final germination on the 10th day. No 100 100 µM Mel 120 µM Mel
germination was recorded in -0.4 MPa (Fig. 1).
80
Effect of melatonin pretreatment on germination parameters
60
under PEG induced drought
40
Germination indicators: The germination percentage (GP) was
observed to be higher in absolute control (AC), attaining its final 20

germination on the 7th day, while a lower GP (49%) was observed 0

in stress control (SC), reaching its final germination on 10th day. 2 4 6 8 10 12
However, seeds treated with different melatonin concentrations Germination time (days)
showed increased germination percentage in shorter germination Fig. 2. Effect of melatonin pretreatment on germination percentage under
time, attaining its final germination on 8th day (Fig. 2). Among PEG-induced drought in tomato on 12th day of stress.

Journal of Applied Horticulture (www.horticultureresearch.net)

322 Anti-stress effects of melatonin pre-treatment on germination indicators in tomato

a 100 b 100
a a Promptness index
200.5 Germination index 1400
Germination stress index ab
Vigour index a a
a

Germinati on stress index (GSI)

b
b b 1100
Germinati on index (GI)

Promptness index (PI)

c c c c
150.5 70 b bc 70
d d c c
e d
f e e
800 f
f d
100.5
g g 40 g 40
500
h

50.5
200
10 10

SC
C

SC

el
el

el

el

el

el
el
el

el

el

el
el

A
A

M
M

M
M

µM

µM

µM

µM

µM

µM
µM

µM
µM

µM

µM

µM

20

40

60

80

0
0
20

0
40

60

80

10

12
10

12

Treatments
Fig. 3. Effect of melatonin pretreatment on germination indicators a) germination index and vigor index and b) promptness index and germination
stress index under PEG-induced drought in tomato on 12th day of stress. The data is presented as the mean of replications and standard error of
means (SEM). The least significant difference test (LSD5%) was computed to compare the means among different treatments. The significance level
was denoted using small letters, given that the means with the same letters are not statistically significant at p = 0.05. (Absolute Control-AC, Stress

a c

and root length stress index (RLSI)

b

Shoot length stress index (SLSI)

Shoot l ength stress inde x

Dry matter stress index (DMSI)

a 150 150
Shoot and root length (cm)

8 a Shoot le ngth b 1.0 0. 10 Root l e ngth s tr ess index

a b a Fresh weight
Root le ngth c Dry weight Dry matter stress index

Dry weight (g)

b a 120
Fresh weight (g)

d b 0.8 0. 08 120 ab
c
6 e bb c c
bc
d d c a a a
f e 0.6 e c 90 d ab 90
g f f 0. 06 b b
g d e c c
4 g e d
0.4 h e 60 e 60
0. 04 d
f f
2 e
0.2 g 0. 02 30 30
f

AC SC 20 40 60 80 100 120 AC SC 20 40 60 80 100 120 AC SC 20 40 60 80 100 120

µM µM µM µM µM µM µM µM µM µM µM µM µM µM µM µM µM µM
Mel Mel Mel Mel Mel Mel Mel Mel Mel Mel Mel Mel Mel Mel Mel Mel Mel Mel
Treatments Treatments Treatments
Fig. 4. Effect of melatonin pretreatment on growth and biomass indicators a) shoot length and root length and b) fresh weight and dry weight and
c) stress indices under PEG-induced drought in tomato on 12th day of stress. The data is presented as the mean of replications and standard error of
means (SEM). The least significant difference test (LSD5%) was computed to compare the means among different treatments. The significance level
was denoted using small letters, given that the means with the same letters are not statistically significant at p = 0.05. (Absolute Control-AC, Stress
Control-SC, Melatonin- Mel).
GP 1.00 The germination index (GI), vigor index (VI), promptness index
GI 0.98 1.00 (PI) and germination stress index (GSI) were significantly (P <
VI 0.98 0.97 1.00 0.05) decreased in stress control (SC) seedlings (Fig. 3a, 3b).
PI 0.99 1.00 0.97 1.00
Among the treatments, 100 µM melatonin pretreatment showed
G SI -0.24 -0.20 -0.18 -0.17 1.00
an increased GI, VI and PI of about 150.5 ± 0.9, 1010.1 ± 16.5,
SL 0.96 0.96 0.99 0.96 -0.17 1.00
and 65 ± 0.4, respectively, that resulted in an increase in GSI
compared to stress control. From the correlation matrix (Fig. 5),
it is observed that GI is highly correlated with GRI (r2=1.00), PI
RL 0.80 0.81 0.88 0.83 0.25 0.88 1.00

(r2=1.00), vigor index with SL (r2=0.99), PI with GRI (r2=1.00)

FW 0.97 0.97 0.96 0.97 -0.37 0.96 0.76 1.00

and GSI with SLSI (r2=0.98) and RLSI (r2=0.98).

DW 0.94 0.93 0.96 0.94 -0.06 0.96 0.89 0.92 1.00

SLSI -0.23 -0.19 -0.14 -0.17 0.98 -0.12 0.30 -0.34 -0.02 1.00

RLSI -0.25 -0.22 -0.16 -0.19 0.98 -0.15 0.30 -0.36 -0.03 0.99 1.00 Shoot and root growth, biomass, and stress indicators: The shoot
DMSI 0.10 0.12 0.18 0.16 0.89 0.18 0.57 -0.02 0.30 0.91 0.89 1.00
length (SL), root length (RL), seedling fresh weight (FW) and
MGT 1.00 0.98 0.97 0.99 -0.22 0.95 0.79 0.96 0.93 -0.22 -0.24 0.12 1.00
dry weight (DW) were found to be significant among treatments
(Fig. 4a, 4b). The results showed that SL, RL, FW and DW were
GRI 0.99 1.00 0.98 1.00 -0.21 0.97 0.82 0.98 0.94 -0.20 -0.23 0.12 0.99 1.00
significantly (P < 0.05) higher in absolute control (AC), while
PI

the lower SL, RL, FW and DW were observed in stress control

CVG 0.76 0.86 0.75 0.85 0.04 0.77 0.72 0.77 0.75 0.02 -0.01 0.24 0.76 0.83 1.00

(SC). Among the treatments, -0.3 MPa (PEG) treated with 100
GP

FW

DW

CVG
MGT
GI

VI

G SI

SLSI

RLSI

DMSI

GRI
SL

RL

Pearson's µM melatonin showed an increased SL (6.28 ± 0.08 cm), RL

correlation
-0.50 -0.25 0 0.25 0.50 0.75 (7.38 ± 0.22 cm), FW (0.66 ± 0.006 mg seedling-1 and DW (0.07
Fig. 5. The correlation matrix between all germination parameters in ± 0.001 mg seedling-1. From the correlation analysis (Fig. 5), it is
tomato was assessed using Pearson correlation at the significance of p
= 0.05. Germination percentage (GP), Germination index (GI), Vigour observed that SL had a significant (P < 0.05) positive correlation
index (VI), Promptness index (PI), Germination stress index (GSI), Shoot with GRI (r2=0.97) and negatively correlated with SLSI (r2=-0.12)
length (SL), Root length (RL), Fresh weight (FW), Dry weight (DW), and RLSI (r2=-0.15). Similarly, RL strongly correlates with DW
Shoot length stress index (SLSI), Root length stress index (RLSI), Dry
matter stress index (DMSI), Mean germination time (MGT), Germination (r2=0.89), while the FW and DW positively correlate with GRI
rate index (GRI) and Coefficient of velocity of germination (CVG). (r2=0.98; r2=0.94), respectively. The shoot length stress index
Journal of Applied Horticulture (www.horticultureresearch.net)
 Anti-stress effects of melatonin pre-treatment on germination indicators in tomato 323

(SLSI), root length stress index (RLSI) and dry matter stress index seed germination capacity. This is due to reduced water absorption
(DMSI) showed significant (P < 0.05) among the treatments and hydrolysis of seed nutrient reserves. Comparable results were
(Fig. 4c). Among the treatments, SLSI, RLSI and DMSI were reported by Kintl et al. (2021) in clover.
minimum in stress control (SC), while the maximum SLSI (86 ±
To reduce the negative effects of drought, one of the
2.9), RLSI (112 ± 2.3) and DMSI (87 ± 4.8) were observed in 100
phytohormones, like melatonin, is widely used and extensively
µM melatonin pre-treated seedlings. In this study, it is observed
reported in rice (Megala et al., 2022) and maize (Ahmad et al.,
that SLSI, RLSI, and DMSI showed a significant (P < 0.05)
2022). However, the role of melatonin on seed germination
positive relationship with RLSI (r2=0.99), DMSI (r2=0.89) and
indicators involved in stress alleviation has rarely been reported.
CVG (r2=0.24), respectively (Fig. 5).
The impact of melatonin on seed germination depends on the
Speed of germination: The rate of germination, germination concentration, indicating that higher concentrations of melatonin
rate index (GRI) and coefÏcient of the velocity of germination may inhibit or not affect germination (Megala et al., 2022).
(CVG) were significantly (P < 0.05) decreased by 49%, 63% However, in our study, melatonin concentration exceeding 100
and 32%, respectively, in stress control (SC) compared to µM slightly declined. The increased negative osmotic potential
absolute control (AC). Among the treatments, the increased significantly affected the germination indicators (Kumar et al.,
rate of germination, GRI, and CVG by 61%, 115% and 41%, 2017), indicating that enzymatic activities have a role in reducing
respectively, were recorded in 100 µM melatonin pre-treated the germination potential. Our study revealed that melatonin
seedlings, while the decreased rate of germination, GRI, and application ameliorated the negative effects, which may be due to
CVG by 22%, 51% and 33%, respectively were recorded in 20 enhanced mobilization of endosperm reserve, energy production
µM melatonin pre-treated seedlings compared to stress control in the respiration process, enzymatic activity, hormonal activity
(Table 2). From the correlation analysis (Fig. 5), it is concluded and dilution of protoplasm (Muscolo et al., 2014). Consequently,
that the speed of germination parameters such as MGT, GRI and more GSI indicates that seedlings withstand drought upon
CVG) is highly correlated with GRI, CVG and GI, respectively, melatonin treatment, showing drought tolerance. Similar studies
having a correlation coefÏcient (r2) value of 0.99, 0.83 and 0.86, were reported comparable results in cucumber (Zhang et al.,
respectively. 2013), and cotton (Bai et al., 2020).
Table 2: Effect of melatonin pretreatment on the speed of germination Crop productivity is assessed by evaluating seedling growth
under PEG-induced drought in tomato indicators such as shoot, root length and biomass indicators,
Treatments Rate of Germination CoefÏcient including fresh and dry weight (Shahzad et al., 2023). Root
germination rate index of velocity of growth is constrained when crops experience drought stress,
(% day-1) (GRI) germination while shoot growth is significantly reduced. The decreased shoot
% day-1 (CVG) length is due to reduced cell elongation and low water potential.
Absolute control 1.90 ± 0.021a 19.04 ± 0.32a 21.58 ± 0.40a Similarly, biomass indicators are also reduced under drought
Stress control 0.96 ± 0.024f 6.98 ± 0.31g 14.58 ± 0.57d stress, indicating turgor loss (George et al., 2013). In the present
study, the application of melatonin enhanced the seedling growth
20 µM Melatonin 1.17 ± 0.034e 10.50 ± 0.22f 19.38 ± 0.47c
and biomass indicators even under limited water availability by
40 µM Melatonin 1.27 ± 0.021d 11.63 ± 0.19e 19.49 ± 0.09bc increasing the absorption capacity of water and nutrients, thus
60 µM Melatonin 1.40 ± 0.021 c
12.83 ± 0.12d
19.84 ± 0.25bc improving biomass. However, it should be noted that although
80 µM Melatonin 1.48 ± 0.040b 13.83 ± 0.20c 20.01 ± 0.43bc these positive results were observed, they were insufÏcient to
compensate for the damage caused by drought fully. Stress indices
100 µM Melatonin 1.54 ± 0.024b 15.04 ± 0.08b 20.59 ± 0.35ab and speed of germination were found to be decreased under
120 µM Melatonin 1.50 ± 0.034b 14.38 ± 0.17c 20.00 ± 0.43bc PEG-induced drought. Similar results were reported in soybean
The data represents the mean of replications and standard error of means by Zou et al. (2019).
(SEM). The least significant difference test (LSD5%) was computed to
compare the means among different treatments. The significance level In contrast, the application of melatonin as pretreatment positively
was denoted using small letters, given that the means with the same influences intercellular pH and cell wall modifications. This
letters are not statistically significant at P = 0.05. resulted in increased shoot length, root length and dry matter
Discussion stress indexes. Comparable results were also reported in
greengram (Anitha et al., 2022). Regarding parameters related
Drought significantly affects seed germination, crop growth,
to germination speed, melatonin treatment led to an increase
development and yield forming process. Germination is a
in rate of germination, substantiating a higher number of seeds
physiological process influenced by regulating various enzymes.
germinated in a day. Additionally, there was an increase in GRI
This process involves imbibition, seedcoat expansion, and
and CVG, representing faster germination in a shorter time. It
enzymatic degradation. In the early stage of the life cycle, such
is essential that CVG only focus on the time required to attain
as germination, polyethylene glycol (PEG) is used to induce
the final germination percentage. Similar studies were reported
drought stress (Khaeim et al., 2022). Hence, the primary goal is
by Bai et al. (2020) . In the current study, correlation with seed
to optimize the PEG concentration before setting an experiment.
germination parameters were found to be positive in most traits
Drought influences seed germination ability, as evidenced in
that, evidences the importance of melatonin pretreatment in
previous studies (Kaur et al., 2023). Our research showed that
mitigating drought stress.
tomato seeds imposed to PEG-induced drought resulted in a
decreased germination percentage, seedling growth and vigor In conclusion, treating tomato seeds with melatonin at varying
index than absolute control. Consequently, higher concentrations concentrations (20-120 μM) improved germination and seedling
of PEG reduced the rate of germination, indicating a decline in growth. Notably, 100 μM melatonin showed the most significant
Journal of Applied Horticulture (www.horticultureresearch.net)
324 Anti-stress effects of melatonin pre-treatment on germination indicators in tomato

enhancements in multiple growth and stress indicators. This study Khaeim, H., Z. Kende, I. Balla, C. Gyuricza, A. Eser and A. Tarnawa,
underscores melatonin's potential as a drought stress elicitor 2022. The effect of temperature and water stresses on seed
germination and seedling growth of wheat (Triticum aestivum L.).
for enhancing commercial-scale yields. However, for broader Sustainability, 14: 3887.
applicability, standardized investigations into melatonin seed
Kintl, A., I. Hunady, T. Vymyslicky, V. Ondriskova, T. Hammerschmiedt,
treatment across different crops and concentrations are essential M. Brtnicky and J. Elbl, 2021. Effect of seed coating and PEG-
for future research. induced drought on the germination capacity of five clover crops.
Plants, 10: 724.
Acknowledgement Kumar, A., N. Reddy and J. Lakshmi, 2017. PEG induced screening
We are grateful to Department of Crop Physiology and for drought tolerance in tomato genotypes. Intl. J. Curr. Microbiol.
Appl. Sci., 6: 168-181.
Department of Vegetable Science, Tamil Nadu Agricultural
Liu, J., W. Wang, L. Wang and Y. Sun, 2015. Exogenous melatonin
University for providing the lab facility and seeds. improves seedling health index and drought tolerance in tomato.
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Received: July, 2023; Revised: September, 2023; Accepted: September, 2023

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