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Original article
Impact of planting density and soaking seeds in melatonin solution on
yield, secondary products content and antimicrobial activity of lovage
plant
Amira K.G. Atteya
a,
⇑
, Aishah N. Albalawi
b
, Hala M. Bayomy
c,d
, El Moataz Bellah A. El-Naggar
e
,
Mahmoud H. Ghozlan
f
, Esmail A.E. Genaidy
g
a
Horticulture Department, Faculty of Agriculture, Damanhour University, Damanhour 22516, Egypt
b
Department of Analytical Chemistry, Tabuk University, University College of Haql, Tabuk 71491, Saudi Arabia
c
Department of Nutrition and Food Science, Tabuk University, Tabuk 71491, Saudi Arabia
d
Department of Food Science and Technology, Damanhour University, Damanhour 22516, Egypt
e
Pharmacognosy Department, Faculty of Pharmacy, Damanhour University, Damanhour 22516, Egypt
f
Plant Pathology Department, Faculty of Agriculture, Damanhour University, Damanhour 22516, Egypt
g
Pomology Department, National Research Centre, Giza 12622, Egypt
article info
Article history:
Received 10 October 2021
Revised 2 November 2021
Accepted 20 December 2021
Available online xxxx
Keywords:
Lovage
Planting density
Melatonin
Essential oil analysis
Total phenolic and antioxidant content
Antimicrobial activity
abstract
Many studies worldwide have been done on the effect of medicinal uses of lovage plant but, very little
works have been done on its production. In this study the effect of different planting density and soaking
seeds in different concentration of melatonin solution as well as their combination treatments on yield,
secondary products content and antimicrobial activity of lovage plant were studied. It was observed that
using planting space of 15 cm gave the maximum mean values of total phenolic and antioxidant content
and essential oil percentage. Using 30 cm planting space gave the maximum mean values of plant height,
yield of herb fresh and dry weight per hectare, yield of roots dry weight and essential oil per hectare.
While the plant space of 45 cm recorded the maximum mean values of fresh and dry weight of herb
and roots fresh weight per plant and chlorophyll content. For melatonin levels, using 100 mM melatonin
solution had the minimum mean values of number of days to emergence. While, soaking seeds in 75 mM
melatonin solution recorded the best results of all studied parameters. Regarding the combination treat-
ments, measurements comprising of herb fresh and dry weight as well as essential oil yield per hectare
showed that the combination treatment of 30 cm between plants in row plus soaking seeds in 75 mM
melatonin solution was able to achieve the maximum values of these parameters. While the combination
treatment of 15 cm between plants in row plus soaking solution of 75 mM melatonin is recommended for
getting the maximum yield of root fresh and dry weight per hectare and the maximum total phenolic and
antioxidant contents per herb in both cuts of both studied season. The first major compound of lovage
essential oil of herb is
a
-terpinyle acetate followed by b-Phellandrene. The percentages of these com-
pounds were affected by the applied treatments. The volatile oil of lovage plant exhibits high antibacte-
rial and antifungal properties in the concentrations range of 75–100 mgmL
1
.
Published by Elsevier B.V. on behalf of King Saud University. This is an open access article under the CC BY
license (http://creativecommons.org/licenses/by/4.0/).
1. Introduction
Levisticum officinale Koch (lovage) is a perennial plant belongs to
family apiaceae. It is a robust, glabrous, perennial herbaceous plant
which grows up to 2–2.5 m. It is native to Southwest Asia and
southern Europe (Tutin et al., 1968). The leaves are 2–3 pinnate,
large and bright green. The flowers are yellowish, the fruits are
5–7 mm long and broad elliptic and the 1000 seed weight is
3.64 g (Heeger, 1956). The main producers are Germany, Hungary,
https://doi.org/10.1016/j.sjbs.2021.12.048
1319-562X/Published by Elsevier B.V. on behalf of King Saud University.
This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Abbreviation: MBC, minimal bactericidal concentration; MFC, minimal fungici-
dal concentration.
⇑
Corresponding author.
E-mail address: amira.khames@agr.dmu.edu.eg (A.K.G. Atteya).
Peer review under responsibility of King Saud University.
Production and hosting by Elsevier
Saudi Journal of Biological Sciences xxx (xxxx) xxx
Contents lists available at ScienceDirect
Saudi Journal of Biological Sciences
journal homepage: www.sciencedirect.com
Please cite this article as: Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al., Impact of planting density and soaking seeds in melatonin solution on yield,
secondary products content and antimicrobial activity of lovage plant, Saudi Journal of Biological Sciences, https://doi.org/10.1016/j.sjbs.2021.12.048
the Netherlands, Poland and Belgium. The traditional usage of
lovage in several diseases has been documented in previous
resources and in folk tradition. Lovage exhibits diverse pharmaco-
logical activities, containing estrogenic, apoptotic and antimy-
cobacterial activities (Bogucka-Kocka et al., 2008; Schinkovitz
et al., 2008). It’s used to treat kidney stones, jaundice, irritable
bowel syndrome, malaria, sore throat, tonsillitis, and cystitis, as
well as rheumatism, gout, boils, and eye inflammation. The bioac-
tivity of lovage oil, and dosages of 40 ppm were found to be effec-
tive in anticancer studies. It is not advised for pregnant women
because it has been linked to the onset of menstruation. People
with kidney illness should avoid using this plant because of its irri-
tating impact, which can cause kidney damage in high dosages .
Lovage plants contain a volatile oil, angelic acid, a bitter extractive,
resins, etc. (Simon et al., 1984; Karnick, 1994). A lot of phytochem-
ical studies have been done to determine the chemical composition
of the essential oil of lovage. The chemical composition of different
extracts of lovage revealed more than 190 volatile compounds, fre-
quently monoterpenes and phtalide (Musitelli and Bossi, 2014). b-
phellandrene,
a
-terpinyl acetate and Zligustilide are the main com-
ponents of lovage oil that present in different value in the various
plant organs. It seemed that, the content and composition of the
essential oil in lovage plant is determined by many factors. Accord-
ing to Samiee et al. (2006) the main constituents in the oil of
lovage, were
a
-terpinyl acetate (40.5%) and b-phellandrene
(16.7%) were the major components. The main components of
the oil in the other report were ß-phellandrene (42.5%), cc-
terpineol (27.9%), cis-ocimene (7.5%) (Reza and Abbas, 2007). All
these finding emphasis on one fact that growth of plants and their
content of bioactive compounds depend on many factors, espe-
cially genetic, physiological and environmental factors. For exam-
ple, the introduction of new crops to cultivate in new areas other
than their habitat enrich the flora of these areas and give different
chemical composition of these new imported plants under the new
environmental conditions which will be suitable for local and
international uses. Meanwhile, the first step for establishing culti-
vation of import plants such as lovage under the new environmen-
tal conditions is finding the best treatments of agricultural
practices to maximizing their production under this new location.
Planting density has a great impact on the yield of plants. Fol-
lowing the optimum density, the highest productivity per unit of
area will be achieved. Where the optimum density works to orga-
nize and facilitate other agricultural practices and regulate compe-
tition between plants over available natural resources such as solar
radiation (Casper and Jackson, 1997; Sharratt and Mcwilliams,
2005; Xu et al., 2008), content of elements, moisture and oxygen
in the soil, as well as the available soil area for growth of roots
(Dong et al., 2010; Ren et al., 2017; Liu et al., 2019). Therefore,
the efficiency of the photosynthesis process will be increased,
which is reflected on the productivity of all different parts of the
plant. This optimum density varies according to the type of plant
and soil, the other agricultural practices as well as the purpose of
production. Many researchers such as Xu et al. (2008) on maize,
Rahman and Hossain (2011) on soybean, Ren et al. (2017), Liu
et al. (2019), Cong et al. (2019) and Tian et al. (2020) on oilseed
rape and Jingang et al. (2019) on sunflower found that growth
and yield of plants were improved with following the optimum
planting density.
As a result of reducing the period between planting and emer-
gence of seeds, the produced seedlings become more strong
(Fenner and Thompson, 2005). As the long period of emergence
expose the seeds to some environmental and microbial hazards
of the soil, such as lack of sufficient moisture, light and oxygen,
as well as attacking micro-organisms. Lovage seeds are character-
ized by a low emergence rate of about 24.67%, as well as the pres-
ence of a dormancy percentage with them (Khodashenas et al.,
2015). This phenomenon of embryo that is not fully developed in
most Apiaceae family (Martin, 1946) has been studied in Apium
graveolens (Pressman et al., 1977) and Conium maculatum (Baskin
and Baskin, 1990). Where the embryo needs appropriate condi-
tions of temperature, humidity, hormones and light to begin in
growth and emergence. Therefore, the soaking of seeds before
planting in melatonin is appropriate treatment for accelerate the
growth of the embryo, which will be reflected on the productivity
of the cultivated plants. Melatonin is a small molecule hormone
found in plants and animals. It is a broad-spectrum growth regula-
tor and antioxidant (Haoshuang et al., 2019). It enhances plant
growth and development especially in abiotic stresses and acts
as an osmotic regulatory substance in plants, enabling the mainte-
nance of ion homeostasis and growth regulation (Timothy and
Colmer, 2015). Previous studies have shown that exogenous mela-
tonin treatments of spraying tea plants with 100 mM, soaking
cucumber seeds in 0.1–500 mM, soaking tomato seedling roots in
1
l
mol L
1
, priming cotton seeds in 10, 20, 50 and 100
l
M mela-
tonin solution (Jiahao et al., 2019; Zhang et al., 2014; Li et al.,
2016; Chen et al., 2020) respectively improved growth under the
condition of normal and abiotic stresses.
Due to the shortage of information about the response of lovage
plant to agriculture practices in general and as a now import plant
under the Egyptian environmental conditions particular, this study
was done to reach the best practices including plant spacing and
melatonin concentration as a soaking treatments of seeds to max-
imize productivity, essential oil, polyphenolics, antioxidant con-
tent, yield and antimicrobial activity of lovage Koch. plant
worldwide and establishment its cultivation under the Egyptian
environmental conditions as a new imported medicinal and aro-
matic plant to try to insert it into Egyptian flora for domestic use
and export.
2. Materials and methods
The present investigation was carried out during two successive
seasons of 2018 / 2019 and 2019 / 2020 in an open field of a private
farm in Kom-Hamada, EL-Behira governorate, Egypt, to study the
effect of plant spacing and soaking seeds in melatonin on growth
as well as essential oil, polyphenolics, antioxidant content, yield
and antimicrobial activity of lovage plants.
2.1. Plant material
Seeds of Levisticum officinale L. (lovage) plant were imported
from Enza Zaden company in North Holland. They were cultivated
on the October of 2018 and 2019. Pest control and other agricul-
tural practices, such as irrigation, etc. were applied, wherever it
was necessary and as commonly recommended. The environmen-
tal conditions of culture location according to Meteorological data
from Central Lab. for Agricultural Climate, Agricultural Research
Center, Ministry of Agriculture and Land Reclamation, Egypt are
shown in the Table 1.
2.2. Treatment
The experiment laid out is a split plot arranged in Randomized
Complete Block Design (RCBD) with three replications during Octo-
ber–March 2018/2019 and 2019/2020. The main plots assigned to
plant spacing and the sub plots deviated to soaking seeds of lovage
plants in melatonin. All possible combination of two studied fac-
tors were done (Table 2). It was taken two cuts of all treatments.
The experiment included 12 treatments which were the combina-
tion between plant spacing (15, 30 and 45 cm) and soaking seeds
before planting for 8 h in different concentration of melatonin
Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al. Saudi Journal of Biological Sciences xxx (xxxx) xxx
2
(without soaking, 0, 75 and 100 mM). Melatonine (N-Acetyl-5-
methoxytryptamine, C
13
H
16
N
2
O
2
) was brought from Sigma-
Aldrich. The experimental unit area was 97.2 m
2
with 13.5 m
length and 7.2 m width and contained twelve rows. The distance
between rows 60 cm apart and between plants in the same row
were 15, 30 or 45 cm. Each main plot contained about 120, 60 or
40 plants under surface irrigation. Seeds were sown on 1st of Octo-
ber in the first and second seasons, respectively. Seeds (three seeds
hill
1
) were sown on one side of the row. After 26 days, the seed-
lings were thinned to one plant per hill. The plots were weeded
every two weeks as it possible. The physical and chemical proper-
ties of the soil samples were determined according to Jackson
(1973) and Cottenie et al. (1982) as shown in Table 3.
2.3. The following data were recorded each season
2.3.1. Emergence parameters
Three 30 cm diameter pots for every melatonin treatment were
planted with soaking seeds (ten seeds per pot). Each pot content
about 10 kg of field soil in an open field conditions. This separated
stage take 20 day from planting until the end of it. As a completely
randomized design (CRD) both of number of days until emergence
(day) and emergence percent (%) were recorded.
2.3.2. Growth and yield parameters
Plant height (cm) from the surface of the soil to the top of the
plant, fresh and dry weight of herb per plant (g), yield of fresh
weight of herb (ton ha
1
) and dry weight of herb (kg ha
1
) were
measured at the end of every cut of every season. While the roots
fresh and dry weight per plant (g) and per hectare (kg ha
1
) were
recorded at the end of every season. The drying of all plant parts
was done by putting them in oven under 48 °C for three days until
getting a constant weight.
2.3.3. Total chlorophyll content (SPAD unit)
It was quantified using a SPAD-502 Chlorophyll Meter (Minolta
Camera Co., Ramsey, NJ).
2.3.4. Total phenolic (mg gallic1 g
1
herb)
It was determined according to the method of Singleton and
Rossi (1965).
2.3.5. Antioxidant content (mg TE 100 g
1
herb)
Free radical scavenging capacity of extracts was determined
using the stable DPPHaccording to Hwang and Do Thi (2014).
The standard curve was prepared using Trolox. Results were
expressed as mg Trolox equivalents (TE) 100 g
1
herb.
2.3.6. Essential oil extraction
Essential oil of lovage were extracted from aerial parts of each
treatment according to Guenther (1961) and as was mentioned
in Atteya and El Gendy (2018) after the harvest in the end of every
cut.
2.3.7. GC–MS analysis
The essential oil of the selected treatments was carried out by
GC–MS analysis as was mentioned in Atteya and El Gendy (2018).
2.3.8. Simple correlation coefficients
Simple correlation coefficients among various studied charac-
ters of lovage in this study as affected with plant spacing and seeds
soaking combination treatments in both seasons.
Table 1
The monthly mean values of the maximum, minimum air temperatures (C°), relative humidity (%), rain (mm/day) and day length (hour) in El-Behiera location during both seasons
of the study.
Date 2018/2019 2019/2020
Temperature
(°C)
Relative humidity % Rain/(mm day) Day Length (h) Temperature
(°C)
Relative humidity Rain/(m day) Day Length (h)
Max Min Max Min
October 28.15 22.80 57.50 0.55 11:33 27.00 20.25 57.20 0.00 11:35
November 24.15 18.05 58.70 1.40 10:55 22.95 16.95 62.30 0.95 10:55
December 18.35 13.60 67.55 0.70 10:35 20.55 15.10 68.30 0.20 10:35
January 17.00 11.05 66.25 0.15 10:48 18.30 12.70 67.45 1.30 10:46
February 17.50 10.95 64.25 0.15 11:20 19.75 12.85 62.15 0.35 11:19
March 21.01 13.06 0.50 0.00 11:56 23.97 14.28 50.18 0.00 11:54
Table 2
All different used combinations treatments of plant spacing and melatonin concen-
trations with the following.
Treatments
T1 15 cm between plants in row and without soaking (control).
T2 15 cm between plants in row and 0 mM melatonin soaking solution.
T3 15 cm between plants in row and 75 mM melatonin soaking solution.
T4 15 cm between plants in row and 100 mM melatonin soaking solution.
T5 30 cm between plants in row and without soaking.
T6 30 cm between plants in row and 0 mM melatonin soaking solution.
T7 30 cm between plants in row and 75 mM melatonin soaking solution.
T8 30 cm between plants in row and 100 mM melatonin soaking solution.
T9 45 cm between plants in row and without soaking.
T10 45 cm between plants in row and 0 mM melatonin soaking solution.
T11 45 cm between plants in row and 75 mM melatonin soaking solution.
T12 45 cm between plants in row and 100 mM melatonin soaking solution.
Table 3
The physical and chemical properties of the experimental soil.
Property Unit Value
Clay % 34
Sand % 27
Silt % 39
Texture class Clay loam
O.M. % 1.4
pH 7.8
ECe ds m
1
2.3
CaCO
3
% 1.8
Soluble ions (meq L
1
)
HCO
3–
2.5
Cl
-
9.3
SO
2-
10.7
Ca
2+
10.9
Mg
2+
6.02
Na
+
4.55
K
+
0.32
Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al. Saudi Journal of Biological Sciences xxx (xxxx) xxx
3
2.3.9. Antimicrobial activity assays
For testing the anti-bacterial and anti-fungal effect of lovage
Koch essential oil, the successive double dilution method was used.
For this, the initial stage, 1 mL of peptone broth medium for tested
bacteria was prepared and the following: phytopathogenic bacteria
of Ralstonia solanacearum,Pectobacterium Carotovorum subsp
atrosepticum and Pectobacterium Carotovorum subsp Carotovorum
were examined. Potato dextrose broth was used as inoculation
medium for the tested fungal strains i.e., Fusarium roseum Subse-
quently, for each bacterial and fungal isolates 1 mL of the obtained
microbial suspension was dropped in a tube containing 9 mL of
sterile distilled water. The content of the tube was mixed, after
which 1 mL was transferred to the tube no. 2 of the 10-tube series
containing 9 mL of sterile distilled water. The procedure was
repeated until the tube no. 10 of the series. Thus, the concentration
of the initial preparation decreased 2-fold in each subsequent tube.
From the 5-th tube of the series were taken 0.1 mL of the microbial
suspension, For the estimation of the minimal used concentration
of essential oil produced from the treatment of 30 cm plant spacing
and seeds soaking solution of 75 mM melatonin as bactericidal and
fungicidal concentrations (MBC, MFC), the contents of the last test
tubes were seeded on peptone Agar medium for determination of
minimal bactericidal effect and Potato dextrose agar medium for
minimal fungicidal effect. The seeded Petri dishes were kept in
the incubator at 25 °C for 24–72 h. The concentration of the tested
preparation that does not allow the growth of any colony of bacte-
rial or fungal colony is considered to be the minimal bactericidal
and fungicidal concentrations of the preparation. According to
Wayne (1999).
2.4. Statistical analysis
The experiment laid out is a split plot arranged in Randomized
Complete Block Design (RCBD) with three replications. Except for
emergence test it was a completely randomized design (CRD).
Analysis of variance with SAS software (SAS Institute Inc., 1988)
was carried out on the all tested treatments data. Treatments’
means were compared using the LSD test at 5% level of probability.
The experiment was repeated in the second year at the same site
using the same steps and techniques of the first year to compare
the results of the two cuts of the two successive seasons.
3. Results
3.1. Effect of melatonin on emergence parameters
Regarding mean values of soaking treatments, results in Table 4
indicate that the three tested treatments had significant effect on
decreasing the number of days until emergence and increasing
emergence percentage as compared with the control in both sea-
sons. Moreover, the minimum number of days until emergence
(5.13 and 6.67 day) was observed with the treatment of soaking
seed in 100 mM melatonin and the maximum emergence percent
(86.67 and 86.67 %) was found with 75 mM melatonin soaking solu-
tion before planting them in both seasons, respectively. From the
other hand the maximum number of days to emergence (13.00
and 13.33 day) as well as the minimum emergence percent
(56.67 and 53.33%) was noticed with control treatment (without
soaking) in both seasons.
3.2. Plant height
Experimental results in Table 5 show that planting density,
melatonin concentration and the interaction of them were able
to make significant differences in the height of lovage plants in
the first and second seasons. Moreover, planting distance of
30 cm between plants in the same row gave the maximum plant
height (36.5 and 28.9 cm) in the first season and (38.1 and
30.8 cm) in the second one for the both cuts, respectively com-
pared with the other used planting spaces. In addition, soaking
seeds in 75 mM melatonin had the highest lovage plants (35.9
and 35.8 cm in the first cut) and (27.7 and 29.1 cm in the second
cut) in both seasons, respectively. As a result of that, the seventh
combined treatment of planting space 30 cm plus 75 mM melatonin
recorded the maximum plant height (38.2 and 30.1 cm in the first
season and 39.4 and 32.6 cm in the second season) in the two cuts.
3.3. Herb fresh weight
Herb fresh weight per plant and per hectare of lovage varied
according to planting space as well as soaking seed treatments
and their interaction. Data in Tables 6 and 7 indicates that despite
the planting at 45 cm distance between plants in the same row
gave the heaviest herb fresh weight per plant (258 and
447 g plant
1
in the first season and 209 and 387 g plant
1
in
the second season, respectively), the maximum yield per hectare
(12.1 and 20.4 ton ha
1
in the first season and 10.0 and
18.3 ton ha
1
in the second season, respectively) was found with
planting distance of 30 cm between plants in the same row com-
pared with the other planting space treatments in the first and sec-
ond cuts. Soaking solution of 75 mM melatonin gave the highest
response to the herb fresh weight in the first season (199 and
353 g plant
1
and 11.3 and 20.0 ton ha
1
) and during the second
season (163 and 312 g plant
1
and 9.2 and 17.3 ton ha
1
) in the
first and second cuts, respectively compared with the other mela-
tonin concentrations. Despite planting at distance 45 cm plus soak-
ing seeds 75 mM melatonin solution recorded the maximum herb
fresh weight per plant (276 and 474 g plant
1
in the first season
and 221 and 434 g plant
1
in the second one) for the two cuts,
respectively the maximum herb fresh weight per hectare (12.6
and 22.9 ton ha
1
in the first season and 10.4 and 19.8 ton ha
1
in the second season) for the both cuts, respectively was found
with the treatment of 30 cm between plants in the same row plus
75 mM melatonin for soaking seeds compared with the other com-
binations treatments.
Table 4
The mean values of number of days to emergence (day) and emergence percent (%) of lovage seeds as affected with melatonin treatments in both seasons.
Concentrations (mM) Number of days to emergence (day) Emergence percent (%)
First season Second season First season Second season
Without soaking 13.00 ± 1.00 a*13.33 ± 0.58 a 56.67 ± 5.77 c 53.33 ± 11.55 c
0 8.93 ± 0.61b 8.33 ± 0.58 b 73.33 ± 5.42 b 73.33 ± 5.76 b
75 6.80 ± 0.80 c 7.33 ± 0.54 bc 86.67 ± 6.11 a 86.67 ± 5.81 a
100 5.13 ± 0.50 d 6.67 ± 0.57 c 76.67 ± 5.67 ab 80.00 ± 0.0 ab
*
Means followed by the same letter are not significantly different by the protected LSD, P 0.05. Data are means value ± SE (n = 3).
Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al. Saudi Journal of Biological Sciences xxx (xxxx) xxx
4
3.4. Herb dry weight
Tables 8 and 9 shows the effect of different planting spaces and
soaking treatments and their interactions on herb dry weight of
lovage plants. The response varied according to the treatments.
Tables 8 and 9 indicated that the 45 cm planting space gave the
highest significant average of herb dry weight per plant (23.6
and 34.0 g plant
1
in the first season and 23.4 and 35.1 g plant
1
in the second season) in the two cuts across all melatonin concen-
trations. On the other hand, 30 cm planting space had the maxi-
mum herb dry weight per hectare (1052 and 1440 kg ha
1
in the
first season and 1122 and 1726 kg ha
1
in the second season) for
both cuts, respectively across all melatonin concentrations. Soak-
ing seeds 75 mM melatonin solution recorded the maximum herb
dry weight per plant (17.9 and 26.7 g plant
1
in the first season
and 18.8 and 28.1 g plant
1
in the second season) as well as max-
imum herb dry weight per hectare (1005 and 1446 kg ha
1
in the
first season and 1044 and 1560 kg ha
1
in the second one) across
all planting space treatments in the first and second cuts, respec-
tively. Using the treatment of 45 cm between plants in the same
Table 5
The mean values of plant height (cm) of lovage plants as affected with plant spacing, melatonin concentrations and their combination treatments in both seasons.
Treatments First season Second season
1
st
cut 2
nd
cut 1
st
cut 2
nd
cut
Planting space between plants in row (cm)
15 30.8 ± 3.0 c*23.1 ± 1.8 b 30.3 ± 2.0 c 21.8 ± 2.4 c
30 36.5 ± 2.2 a 28.9 ± 1.7 a 38.1 ± 1.5 a 30.8 ± 2.6 a
45 34.0 ± 2.6 b 27.0 ± 1.8 a 35.0 ± 1.3 b 28.9 ± 1.9 b
Melatonin concentrations (mM)
Without soaking 30.1 ± 3.0 c 24.6 ± 2.7 c 32.7 ± 4.2 c 24.7 ± 3.7 c
0 33.9 ± 3.0 b 26.3 ± 3.3 b 34.6 ± 2.7 b 27.0 ± 4.6 b
75 35.9 ± 2.1 a 27.7 ± 2.6 a 35.8 ± 3.4 a 29.1 ± 5.7 a
100 35.2 ± 2.9 ab 26.8 ± 2.8 ab 34.8 ± 3.9 b 27.8 ± 3.1 b
Interaction
T1 26.7 ± 0.6 f 21.5 ± 1.8 h 27.7 ± 1.5 h 20.0 ± 1.0 e
T2 30.5 ± 2.2 e 22.7 ± 1.7gh 31.7 ± 0.3 fg 21.3 ± 1.0e
T3 33.8 ± 1.4 bcd 24.6 ± 1.1 efg 31.8 ± 0.8 fg 20.3 ± 0.3 e
T4 32.2 ± 1.0 de 23.8 ± 1.7 fgh 30.2 ± 1.4 g 25.6 ± 0.6 d
T5 33.3 ± 0.8 cd 26.3 ± 0.8 def 36.7 ± 1.8 bcd 27.0 ± 1.8 cd
T6 36.5 ± 1.2 ab 29.4 ± 0.8 abc 37.7 ± 1.3 abc 31.4 ± 0.9 ab
T7 38.2 ± 1.2 a 30.1 ± 1.0 a 39.4 ± 0.8 a 32.6 ± 0.8 a
T8 37.8 ± 1.5 a 29.8 ± 0.7 ab 38.8 ± 1.2 ab 32.1 ± 1.4 a
T9 30.3 ± 1.2 e 25.8 ± 1.9 def 33.9 ± 0.4 ef 27.0 ± 1.5 cd
T10 34.7 ± 1.2 bcd 26.7 ± 2.6 cdef 34.4 ± 1.3 de 28.2 ± 1.4 c
T11 35.7 ± 0.6 abc 28.5 ± 0.5 abcd 36.2 ± 1.1 cd 31.0 ± 1.0 ab
T12 35.5 ± 2.3 abc 27.0 ± 1.0 bcde 35.3 ± 1.0 de 29.3 ± 1.1 bc
*
Means followed by the same letter are not significantly different by the protected LSD, P 0.05. Data are means value ± SE (n= 3). 1st cut and 2nd cut means the first and
second cutts, respectively.
Table 6
The mean values of herb fresh weight (g plant
1
) of lovage plants as affected with plant spacing, melatonin concentrations and their combination treatments in two cuts of both
seasons.
Treatments First season Second season
1
st
cut 2
nd
cut 1
st
cut 2
nd
cut
Planting space between plants in row (cm)
15 85 ± 8.1 c*145 ± 23.9 c 73 ± 6.9 c 128 ± 13.7 c
30 218 ± 9.1 b 367 ± 38.0 b 180 ± 4.6 b 330 ± 21.8 b
45 258 ± 12.5 a 447 ± 18.8 a 209 ± 9.9 a 387 ± 30.4 a
Melatonin concentrations (mM)
Without soaking 176 ± 76.5 d 288 ± 138.3 d 148 ± 62.8 c 256 ± 112.6 c
0 183 ± 78.6 c 305 ± 134.2 c 152 ± 61.5 cb 279 ± 114.7 b
75 199 ± 81.5 a 353 ± 136.5 a 163 ± 62.4 a 312 ± 129.6 a
100 190 ± 77.5 b 332 ± 135.2 b 155 ± 62.2 b 278 ± 115.0 b
Interaction
T1 77 ± 7.7 i 114 ± 3.5 k 65 ± 3.9 f 110 ± 5.8 i
T2 81 ± 3.8 hi 135 ± 3.1 j 72 ± 1.7 f 128 ± 6.3 h
T3 94 ± 1.9 g 175 ± 0.8 h 83 ± 2.0 e 145 ± 4.8 g
T4 89 ± 1.4 gh 155 ± 1.8 i 74 ± 1.0 ef 129 ± 1.8 h
T5 205 ± 3.0 f 324 ± 10.3 g 177 ± 1.9 d 301 ± 2.8 f
T6 216 ± 2.1 e 344 ± 6.7f 179 ± 3.6 d 337 ± 12.1 d
T7 226 ± 2.8 d 412 ± 2.0 d 186 ± 2.3 d 357 ± 3.5 c
T8 224 ± 1.2 de 390 ± 1.3 e 179 ± 4.6 d 324 ± 5.0 e
T9 246 ± 1.0 c 426 ± 3.3 cd 201 ± 2.5 c 357 ± 8.2 c
T10 253 ± 2.7 bc 437 ± 2.0 bc 205 ± 8.1 bc 373 ± 5.4 b
T11 276 ± 4.6 a 474 ± 3.0 a 221 ± 1.0 a 434 ± 1.6 a
T12 258 ± 8.4 b 450 ± 4.6 b 211 ± 10.9 b 382 ± 6.7 b
*
Means followed by the same letter are not significantly different by the protected LSD, P 0.05. Data are means value ± SE (n = 3). 1st cut and 2nd cut means the first and
second cutts, respectively.
Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al. Saudi Journal of Biological Sciences xxx (xxxx) xxx
5
row plus 75 mM melatonin gave the maximum herb dry weight per
plant (24.7 and 38.7 g plant
1
in the first season and 25.9 and
38.4 g plant
1
in the second season) compared with the other com-
bination treatments in both cuts, respectively. From the other
hand, the combination treatment of 30 cm between plants in the
same row plus 75 mM melatonin recorded the heaviest herb dry
weight per hectare (1127 and 1687 kg ha
1
in the first season
and 1214 and 1852 kg ha
1
in the second season) for the first
and second cuts, respectively.
3.5. Root fresh and dry weights
Roots fresh and dry weights per plant and yield per hectare
which were determined for lovage plants depending on studied
treatments are shown in Tables 10 and 11. Results showed that
plant spacing, seeds soaking and their combination treatments
are significant factors that affecting roots fresh and dry weights
per plant and per hectare in the first and second seasons. Cultiva-
tion of lovage seeds at 45 cm planting space had the maximum
Table 7
The mean values of herb fresh weight (ton ha
1
) of lovage plants as affected with plant spacing, melatonin concentrations and their combination treatments in two cuts of both
seasons.
Treatments First season Second season
1
st
cut 2
nd
cut 1
st
cut 2
nd
cut
Planting space between plants in row (cm)
15 9.5 ± 0.38 b*16.1 ± 1.12 b 8.2 ± 0.32 b 14.2 ± 0.64 b
30 12.1 ± 0.21 a 20.4 ± 0.89 a 10.0 ± 0.11 a 18.3 ± 0.51 a
45 9.6 ± 0.19 b 16.5 ± 0.29 b 7.8 ± 0.15 c 14.3 ± 0.47 b
Melatonin concentrations (mM)
Without soaking 9.7 ± 0.57 d 15.5 ± 1.02 d 8.2 ± 0.54 c 14.1 ± 0.88 c
0 10.1 ± 0.60 c 16.8 ± 0.78 c 8.5 ± 0.47 b 15.6 ± 1.01 b
75 11.3 ± 0.47 a 20.0 ± 0.98 a 9.2 ± 0.40 a 17.3 ± 0.79 a
100 10.6 ± 0.58 b 18.5 ± 1.00 b 8.7 ± 0.42 b 15.5 ± 0.79 b
Interaction
T1 8.5 ± 0.85 f 12.6 ± 0.39 l 7.2 ± 0.44 g 12.2 ± 0.64 f
T2 9.0 ± 0.43 ef 15.0 ± 0.35 k 8.0 ± 0.18 de 14.3 ± 0.71 d
T3 10.5 ± 0.22 c 19.4 ± 0.09c 9.2 ± 0.22 c 16.1 ± 0.53 c
T4 9.9 ± 0.16 cd 17.2 ± 0.20 g 8.2 ± 0.11 d 14.3 ± 0.19 d
T5 11.4 ± 0.17 b 18.0 ± 1.29 e 9.8 ± 0.10 b 16.8 ± 0.15 c
T6 12.0 ± 0.13 ab 19.1 ± 0.37 d 9.9 ± 0.21 ab 18.7 ± 0.67 b
T7 12.6 ± 0.15 a 22.9 ± 0.12 a 10.4 ± 0.13 a 19.8 ± 0.19 a
T8 12.5 ± 0.06 a 21.7 ± 0.07 b 10.0 ± 0.27 ab 18.0 ± 0.27 b
T9 9.1 ± 0.04 ef 15.8 ± 0.12 j 7.4 ± 0.10 fg 13.2 ± 0.31 e
T10 9.4 ± 0.10 de 16.2 ± 0.09 i 7.6 ± 0.30 efg 13.8 ± 0.21 de
T11 10.2 ± 0.17 c 17.5 ± 0.12 f 8.2 ± 0.04 d 16.1 ± 0.06 c
T12 9.5 ± 0.31 de 16.7 ± 0.17 h 7.8 ± 0.41 def 14.2 ± 0.25 d
*
Means followed by the same letter are not significantly different by the protected LSD, P 0.05. Data are means value ± SE (n = 3). 1st cut and 2nd cut means the first and
second cutts, respectively.
Table 8
The mean values of herb dry weight (g plant
1
) of lovage plants as affected with plant spacing, melatonin concentrations and their combination treatments in two cuts of both
seasons.
Treatments First season Second season
1
st
cut 2
nd
cut 1
st
cut 2
nd
cut
Planting space between plants in row (cm)
15 7.7 ± 0.8 c*9.7 ± 1.6 c 7.6 ± 0.9 c 11.7 ± 1.4 c
30 18.9 ± 1.6 b 25.9 ± 3.6 b 20.2 ± 1.5 b 31.1 ± 2.9 b
45 23.6 ± 1.1 a 34.0 ± 3.8 a 23.4 ± 1.9 a 35.1 ± 3.2 a
Melatonin concentrations (mM)
Without soaking 15.7 ± 6.8 c 20.4 ± 10.3 c 15.8 ± 6.6 c 22.6 ± 9.9 c
0 16.4 ± 7.1 bc 21.9 ± 10.2 bc 16.4 ± 6.9 c 25.6 ± 10.3 b
75 17.9 ± 7.2 a 26.7 ± 12.3 a 18.8 ± 7.8 a 28.1 ± 11.9 a
100 17.0 ± 7.4 ab 24.0 ± 10.6 b 17.3 ± 7.7 b 27.5 ± 11.4 a
Interaction
T1 7.1 ± 0.3 e 7.5 ± 0.3 f 7.1 ± 0.9 f 9.8 ± 0.9 e
T2 7.5 ± 0.5 e 9.6 ± 0.5 f 7.5 ± 0.5 f 11.8 ± 0.3 de
T3 8.8 ± 0.4 e 11.0 ± 0.7 f 8.6 ± 0.7 f 12.6 ± 1.0 d
T4 7.5 ± 0.8 e 10.9 ± 1.2 f 7.2 ± 0.5 f 12.6 ± 0.9 d
T5 17.9 ± 0.3 d 23.7 ± 2.1 e 18.8 ± 0.4 e 26.7 ± 0.6 c
T6 18.4 ± 2.3 d 23.3 ± 3.1 e 19.3 ± 1.5 de 31.3 ± 0.6 b
T7 20.3 ± 1.5 c 30.4 ± 1.0 cd 21.9 ± 0.7 c 33.3 ± 1.5 b
T8 19.2 ± 1.2 cd 26.3 ± 3.2 de 20.9 ± 0.8 cd 32.9 ± 0.9 b
T9 22.2 ± 0.6 b 30.0 ± 4.4 cd 21.4 ± 0.8 c 31.3 ± 2.1 b
T10 23.4 ± 0.4 ab 32.7 ± 0.6 bc 22.4 ± 1.2 bc 33.5 ± 0.5 b
T11 24.7 ± 0.8 a 38.7 ± 1.2 a 25.9 ± 0.5 a 38.4 ± 1.6 a
T12 24.1 ± 0.4 a 34.7 ± 0.6 ab 23.9 ± 0.6 b 37.0 ± 1.1 a
*
Means followed by the same letter are not significantly different by the protected LSD, P 0.05. Data are means value ± SE (n = 3). 1st cut and 2nd cut means the first and
second cutts, respectively.
Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al. Saudi Journal of Biological Sciences xxx (xxxx) xxx
6
significant root fresh weight (135.5 and 143.8 g plant
1
) and roots
dry weight per plant (13.6 and 13.7 g plant
1
) in both studied sea-
sons compared with the other studied planting space. On the con-
trary, the maximum yield of roots fresh weight (7742 and
7565 kg ha
1
) in both seasons were recorded with sowing seeds
under 15 cm planting space conditions, while the maximum signif-
icant yield of roots dry weight per hectare (633 and 623 kg ha
1
)in
the first and second seasons were found with plants sown in 30 cm
planting space compared with the other studied planting space. By
comparing the melatonin concentrations, the application of 75 mM
melatonin solution for soaking seeds of lovage was able to achieve
the maximum significant roots fresh weight per plant (112.5 and
116.1 g plant
1
) and per hectare (6709 and 6829 kg ha
1
), roots
dry weight per plant (10.9 and 10.6 g plant
1
) and per hectare
(624 and 621 kg ha
1
) in both studied seasons, respectively com-
pared with the other studied soaking period of seeds. For the com-
bination treatments of planting spaces and melatonin
concentrations, the eleven combination treatment of 45 cm
Table 9
The mean values of herb dry weight (kg ha
1
) of lovage plants as affected with plant spacing, melatonin concentrations and their combination treatments in two cuts of both
seasons.
Treatments First season Second season
1
st
cut 2
nd
cut 1
st
cut 2
nd
cut
Planting space between plants in row (cm)
15 858 ± 37 b*1082 ± 74 c 847 ± 40 b 1302 ± 65 b
30 1052 ± 37 a 1440 ± 84 a 1122 ± 36 a 1726 ± 67 a
45 875 ± 17 b 1259 ± 60 b 866 ± 29 b 1298 ± 49 b
Melatonin concentrations (mM)
Without soaking 868 ± 41 c 1087 ± 97 c 875 ± 57 c 1244 ± 81 c
0 908 ± 46 bc 1191 ± 59 c 912 ± 55 bc 1433 ± 98 b
75 1005 ± 45 a 1446 ± 88 a 1044 ± 57 a 1560 ± 98 a
100 933 ± 49 b 1319 ± 67 b 950 ± 70 b 1532 ± 97 a
Interaction
T1 786 ± 28 g 836 ± 31 f 789 ± 102 e 1090 ± 10 e
T2 836 ± 50 fg 1067 ± 53 e 836 ± 50 e 1316 ± 31 e
T3 972 ± 44 bcde 1219 ± 80 cde 960 ± 81 cd 1404 ± 111 cd
T4 838 ± 92 fg 1209 ± 134 de 803 ± 52 e 1397 ± 103 de
T5 994 ± 18 bcd 1315 ± 116 bcd 1042 ± 21 c 1481 ± 32 c
T6 1022 ± 126 abc 1296 ± 170 bcd 1069 ± 85 bc 1741 ± 32 bc
T7 1127 ± 81 a 1687 ± 56 a 1214 ± 37 a 1852 ± 85 a
T8 1065 ± 69 ab 1463 ± 179 b 1163 ± 47 ab 1830 ± 52 ab
T9 823 ± 24 fg 1111 ± 161 de 793 ± 31 e 1160 ± 77 e
T10 867 ± 16 efg 1210 ± 21de 831 ± 44 e 1242 ± 19 e
T11 915 ± 30 cdef 1432 ± 43 bc 957 ± 18 cd 1422 ± 61 cd
T12 894 ± 15defg 1284 ± 21 bcd 884 ± 22 de 1369 ± 39 e
*
Means followed by the same letter are not significantly different by the protected LSD, P 0.05. Data are means value ± SE (n = 3). 1st cut and 2nd cut means the first and
second cutts, respectively.
Table 10
The mean values of roots fresh weight (g plant
1
)and yield of roots fresh weight (kg ha
1
) of lovage plants as affected with plant spacing, melatonin concentrations and their
combination treatments in two cuts of both seasons.
Treatments Roots fresh weight (g plant
1
) Roots fresh weight (kg ha
1
)
First season Second season First season Second season
Planting space between plants in row (cm)
15 69.7 ± 1.5 c*68.1 ± 2.0 c 7742 ± 71 a 7565 ± 94 a
30 124.8 ± 2.2 b 126.2 ± 2.1 b 6933 ± 51 b 7009 ± 49 b
45 135.5 ± 2.8 a 143.8 ± 5.1 a 5019 ± 43 c 5326 ± 80 c
Melatonin concentrations (mM)
Without soaking 107.7 ± 30.3 c 109.4 ± 33.5 d 6418 ± 491c 6438 ± 420 d
0 109.6 ± 30.4 b 111.2 ± 33.2 c 6548 ± 515 b 6569 ± 446 c
75 112.5 ± 31.3 a 116.1 ± 35.6 a 6709 ± 514 a 6829 ± 426 a
100 110.2 ± 30.5 b 114.0 ± 35.2 b 6584 ± 517 b 6697 ± 413 b
Interaction
T1 67.8 ± 1.0 f 65.5 ± 1.1 f 7533 ± 110 c 7280 ± 125 c
T2 69.5 ± 0.9 ef 67.9 ± 0.4 ef 7725 ± 97b 7539 ± 48 b
T3 71.3 ± 1.1 e 70.3 ± 1.0 e 7927 ± 126 a 7808 ± 113 a
T4 70.1 ± 0.2 ef 68.7 ± 1.4 e 7783 ± 21 ab 7633 ± 157 ab
T5 122.4 ± 2.4 d 124.3 ± 0.7 d 6800 ± 131 e 6906 ± 39 e
T6 125.0 ± 1.4 cd 125.5 ± 1.7 d 6943 ± 75 de 6974 ± 97 de
T7 126.7 ± 1.9 c 128.6 ± 0.5 c 7037 ± 107 d 7144 ± 29 cd
T8 125.2 ± 0.8 cd 126.3 ± 2.4 cd 6954 ± 42 de 7014 ± 133 de
T9 132.9 ± 0.7 b 138.4 ± 1.8 b 4921 ± 25 g 5127 ± 68 g
T10 134.4 ± 1.8 b 140.3 ± 2.2 b 4978 ± 68 g 5195 ± 83 g
T11 139.4 ± 0.6 a 149.5 ± 2.5 a 5162 ± 24 f 5536 ± 91f
T12 135.4 ± 2.0 b 147.0 ± 2.1 a 5014 ± 73 fg 5444 ± 78f
*
Means followed by the same letter are not significantly different by the protected LSD, P 0.05. Data are means value ± SE (n = 3). 1st cut and 2nd cut means the first and
second cutts, respectively.
Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al. Saudi Journal of Biological Sciences xxx (xxxx) xxx
7
between plants in row plus using 75 mM melatonin solution before
sowing recoded the maximum roots fresh weight (139.4 and
149.5 g plant
1
) and roots dry weight per plant (14.6 and 14.0 g
plant
1
) in both studied seasons, respectively. However, the maxi-
mum yield of roots fresh weight (7927 and 7808 kg ha
1
) in both
seasons and roots dry weight per hectare (702 kg ha
1
) in the sec-
ond one were recorded with the third combination treatment of
15 cm between plants in row plus soaking seeds in 75 mM mela-
tonin solution before sowing seeds but the maximum yield of roots
dry weight per hectare (669 kg ha
1
) in the first season was
noticed with plants of the seventh combination treatment of
30 cm between plants in row plus using 75 mM melatonin before
the sowing compared with the other studied combination
treatments.
3.6. Chlorophyll, total phenolic and antioxidant content
The results of chlorophyll content, total phenolic and antioxi-
dant content means comparison of lovage plants as affected with
planting spacing, melatonin concentrations and their combination
treatments are shown in Tables 12, 13 and 14. The highest means
values of chlorophyll content (49.69 and 52.49 spad unit in the first
season and 51.58 and 48.40 spad unit in the second season) for
both cuts, respectively were noticed with planting distance of
45 cm between plants in the same row with insignificant differ-
ences between the means values in the second cut of the second
season. While, The maximum means values of total phenolic
(14.28 and 13.54 mg gallic 1 g
1
herb in the first season and
14.25 and 13.90 mg gallic 1 g
1
herb in the second season) and
antioxidant content (53.94 and 54.49 mg TE 100 g
1
herb in the
first season and 56.63 and 55.57 mg TE 100 g
1
herb in the second
season) for studied cuts, respectively was observed in lovage plants
sown in 15 cm planting spaces conditions compared with the other
different studied planting spaces in both seasons, respectively.
Based on melatonin application, soaking lovage seeds before sow-
ing 75 mM melatonin solution gave the highest significant means
values of chlorophyll content (48.88 and 49.37 spad unit in the first
cut and 50.61 spad unit in the second cut), total phenolic (12.98
and 12.80 mg gallic 1 g
1
herb in the first cut and 12.39 and
12.59 mg gallic 1 g
1
herb in the second cut) and antioxidant con-
tent (50.11 and 51.66 mg TE 100 g
1
herb in the first cut and 49.57
and 50.57 mg TE 100 g
1
herb in the second cut) in both seasons
except the maximum significant means value of chlorophyll con-
tent (52.06 spad unit) in the second cut of the second season were
found with melatonin concentration of 100 mM compared with the
other studied concentrations. Regarding the effect of combination
treatments, the combination treatment of 45 cm between plants
in row plus 75 mM melatonin solution gave the maximum values
of chlorophyll content (52.00 and 56.27 spad unit) and (52.37
and 55.03 spad unit) in both cuts of the two seasons. Moreover,
the combination treatment of T3 (15 cm between plants in row
and 75 mM melatonin) had the maximum mean values of total phe-
nolic (14.96 and 14.10 mg gallic 1 g
1
herb in the first season and
14.62 and 14.27 mg gallic 1 g
1
herb in the second season) and
antioxidant content (54.32 and 55.33 mg TE 100 g
1
herb in the
first season and 57.79 and 56.95 mg TE 100 g
1
herb in the second
season) for the first and the second cuts, respectively.
3.7. Essential oil content
Tables 15, 16 and 17 shows the ability of plant spacing, soaking
seeds and their interaction to induce significant differences in the
essential oil percentage, essential oil per plant and per hectare in
the first and second seasons and in both cuts. Despite the highest
essential oil percentage of lovage plants (0.568 and 0.455% in the
first season and 0.569 and 0.494% in the second season) that
recorded with using the closer spacing (15 cm), the maximum
essential oil per plant (0.101 mL plant
1
in the first season) in
the first cut and (0.113 and 0.142 mL plant
1
in the second season)
in both cuts and also, per hectare (5.61 and 7.52 L ha
1
in the first
season and 6.30 and 7.87 L ha
1
in the second season) for the first
and second cuts, respectively were found with using medium
space of 30 cm. Both of 30 and 45 cm between plants had the max-
imum significant essential oil content per plant in the second cut of
Table 11
The mean values of roots dry weight (g plant
1
) and roots dry weight (kg ha
1
) of lovage plants as affected with plant spacing, melatonin concentrations and their combination
treatments in two cuts of both seasons.
Treatments Roots dr y weight (g plant
1
) Roots dry weight (kg ha
1
)
First season Second season First season Second season
Planting space between plants in row (cm)
15 5.3 ± 0.5 c*5.5 ± 1.1 c 583 ± 61 b 589 ± 76 b
30 11.4 ± 0.7 b 11.2 ± 0.6 b 633 ± 40 a 623 ± 32 a
45 13.6 ± 0.7 a 13.7 ± 0.4 a 503 ± 28 c 506 ± 16 c
Melatonin concentrations (mM)
Without soaking 9.4 ± 3.7 d 9.5 ± 2.4 c 526 ± 55 d 535 ± 48 c
0 9.8 ± 3.7 c 9.9 ± 2.5 bc 558 ± 56 c 555 ± 58 bc
75 10.9 ± 3.9 a 10.6 ± 1.6 a 624 ± 63 a 621 ± 83 a
100 10.3 ± 3.8 b 10.2 ± 2.1 b 584 ± 71 b 580 ± 58 b
Interaction
T1 4.6 ± 0.2 h 4.7 ± 0.3 e 514 ± 26 fg 522 ± 28 d
T2 5.1 ± 0.2 h 4.8 ± 0.4 e 564 ± 20 de 538 ± 4 5 d
T3 6.0 ± 0.2 g 6.3 ± 0.5 d 661 ± 24 ab 702 ± 50 a
T4 5.3 ± 0.4 gh 5.4 ± 0.2 e 593 ± 41 cd 596 ± 23 c
T5 10.6 ± 0.5 f 10.6 ± 0.7 c 591 ± 27 cd 589 ± 38 c
T6 11.1 ± 0.4 ef 11.2 ± 0.6 bc 617 ± 20 bc 624 ± 32 bc
T7 12.0 ± 0.3 d 11.6 ± 0.3 b 669 ± 14 a 643 ± 14 b
T8 11.8 ± 0.7 de 11.4 ± 0.4 b 656 ± 39 ab 635 ± 22 bc
T9 12.8 ± 0.3 c 13.3 ± 0.0 a 473 ± 13 g 493 ± 0 d
T10 13.3 ± 0.3 bc 13.6 ± 0.4 a 493 ± 13 g 505 ± 15 d
T11 14.6 ± 0.3 a 14.0 ± 0.6 a 542 ± 10 ef 519 ± 23 d
T12 13.6 ± 0.2 b 13.7 ± 0.2 a 504 ± 8 fg 509 ± 8 d
*
Means followed by the same letter are not significantly different by the protected LSD, P 0.05. Data are means value ± SE (n = 3). 1st cut and 2nd cut means the first and
second cutts, respectively.
Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al. Saudi Journal of Biological Sciences xxx (xxxx) xxx
8
the first season. By comparing the used melatonin concentrations,
the application of 75 mM melatonin concentration gave the maxi-
mum essential oil percentage, essential oil content per plant and
per hectare (0.559 and 0.447%, 0.095 and 0.115 mL plant
1
and
5.66 and 6.50 L ha
1
) in the first season and (0.597 and 0.503%,
0.106 and 0.134 mL plant
1
and 6.30 and 7.84 L ha
1
) in the second
season for both cuts, respectively compared with the other mela-
tonin concentrations. Insignificant differences were detected
between 0 and 75 mM melatonin concentrations in the second
cut of the first season for essential oil yield per hectare. Further-
more, the results obtained indicate that the greatest essential oil
percent (0.629 and 0.495% in the first season and 0.687 and
0.578% in the second season) for both cuts, respectively were
obtained with combination treatment of plant spacing 15 cm plus
75 mM melatonin. While, the maximum essential oil content per
plant (0.124 mL plant
1
in the first cut of the first season and
Table 12
The mean values of chlorophyll content (SPAD unit) of lovage plants as affected with planting spacing, melatonin concentrations and their combination treatments in two cuts of
both seasons.
Treatments First season Second season
1
st
cut 2
nd
cut 1
st
cut 2
nd
cut
Planting space between plants in row (cm)
15 44.90 ± 2.81 b*42.83 ± 2.32 c 42.17 ± 2.36 c 46.48 ± 4.04 a
30 45.98 ± 1.69 b 48.30 ± 1.92 b 49.34 ± 1.85 b 47.27 ± 4.91 a
45 49.69 ± 2.46 a 52.49 ± 2.58 a 51.58 ± 1.43 a 48.40 ± 3.43 a
Melatonin concentrations (mM)
Without soaking 44.42 ± 3.03 c 45.99 ± 5.03 c 45.70 ± 5.22 c 44.40 ± 3.79 c
0 46.00 ± 2.20 b 47.73 ± 4.16 b 47.70 ± 3.88 b 45.49 ± 2.99 c
75 48.88 ± 2.59 a 50.61 ± 4.85 a 49.37 ± 3.44 a 48.92 ± 1.81 b
100 48.13 ± 2.72 a 47.16 ± 3.61 bc 48.01 ± 5.06 b 52.06 ± 2.81 a
Interaction
T1 41.43 ± 1.00 e 39.80 ± 1.45 h 39.27 ± 0.68 g 41.67 ± 0.47 d
T2 43.83 ± 1.85 de 42.50 ± 1.75 g 42.83 ± 0.32 ef 44.90 ± 4.09 cd
T3 47.40 ± 0.60 c 45.23 ± 0.38 ef 45.07 ± 0.81 de 49.43 ± 1.76 b
T4 46.93 ± 1.85 cd 43.77 ± 0.68 fg 41.50 ± 1.68 fg 49.93 ± 0.59 b
T5 43.97 ± 1.85 de 47.47 ± 1.24 de 47.10 ± 1.71 cd 42.13 ± 0.76 d
T6 46.07 ± 1.32 cd 49.40 ± 0.66 cd 49.20 ± 0.56 bc 48.03 ± 1.45 bc
T7 47.23 ± 0.70 c 50.33 ± 0.72 bc 50.67 ± 1.53 ab 48.40 ± 1.97 bc
T8 46.67 ± 0.95 cd 46.00 ± 0.90 ef 50.40 ± 1.22 ab 48.93 ± 2.30 b
T9 47.87 ± 0.92 bc 50.70 ± 1.91 bc 50.73 ± 1.72 ab 49.40 ± 0.44 b
T10 48.10 ± 0.70 bc 51.30 ± 1.11 bc 51.07 ± 2.00 ab 43.53 ± 1.01 d
T11 52.00 ± 2.00 a 56.27 ± 1.42 a 52.37 ± 0.91 a 55.03 ± 0.84 a
T12 50.80 ± 3.04 ab 51.70 ± 0.72 b 52.13 ± 0.76 a 51.2 ± 3.06 b
*
Means followed by the same letter are not significantly different by the protected LSD, P 0.05. Data are means value ± SE (n = 3). 1st cut and 2nd cut means the first and
second cutts, respectively.
Table 13
The mean values of total phenolic (mg gallic 1 g
1
herb) of lovage plants as affected with planting spacing, melatonin concentrations and their combination treatments in two cuts
of both seasons.
Treatments First season Second season
1
st
cut 2
nd
cut 1
st
cut 2
nd
cut
Planting space between plants in row (cm)
15 14.28 ± 0.55 a*13.54 ± 0.59 a 14.25 ± 0.51 a 113.90 ± 0.35 a
30 12.18 ± 0.64 b 11.62 ± 0.55b 12.20 ± 0.58 b 111.94 ± 0.63 b
45 10.20 ± 0.73 c 9.62 ± 0.72c 110.19 ± 0.66 c 110.00 ± 0.71 c
Melatonin concentrations (mM)
Without soaking 11.54 ± 1.92 d 10.86 ± 1.76 d 11.67 ± 1.85 b 11.31 ± 1.89 d
0 11.99 ± 1.91 c 11.44 ± 1.72c 11.85 ± 1.92 b 11.76 ± 1.88 c
75 12.98 ± 1.69 a 12.39 ± 1.52 a 12.80 ± 1.58 a 12.59 ± 1.46 a
100 12.37 ± 1.66 b 11.68 ± 1.84b 12.52 ± 1.79 a 12.13 ± 1.63 b
Interaction
T1 13.80 ± 0.12 b 12.73 ± 0.32c 13.81 ± 0.37 b 13.53 ± 0.12 b
T2 14.11 ± 0.06 b 13.43 ± 0.21b 14.04 ± 0.21 ab 13.83 ± 0.06 ab
T3 14.96 ± 0.62 a 14.10 ± 0.26 a 14.62 ± 0.68 a 14.27 ± 0.29 a
T4 14.25 ± 0.43 b 13.90 ± 0.10 a 14.52 ± 0.36 a 13.97 ± 0.42 ab
T5 11.40 ± 0.34 e 11.12 ± 0.08 d 11.61 ± 0.29 c 11.17 ± 0.33 e
T6 12.12 ± 0.50 d 11.42 ± 0.21 d 11.84 ± 0.55 d 11.88 ± 0.49 d
T7 12.82 ± 0.26 c 12.47 ± 0.15c 12.71 ± 0.33 c 12.57 ± 0.25 c
T8 12.38 ± 0.48 cd 11.47 ± 0.15 d 12.62 ± 0.12 c 12.13 ± 0.47 cd
T9 9.40 ± 0.37 g 8.72 ± 0.12 g 9.58 ± 0.09 g 9.22 ± 0.36 g
T10 9.74 ± 0.20 g 9.48 ± 0.12 f 9.67 ± 0.25 g 9.55 ± 0.19 g
T11 11.15 ± 0.05 ef 10.61 ± 0.16 e 11.09 ± 0.17 e 10.93 ± 0.05 e
T12 10.49 ± 0.09 f 9.68 ± 0.25 f 10.41 ± 0.16f 10.29 ± 0.09 f
*
Means followed by the same letter are not significantly different by the protected LSD, P 0.05. Data are means value ± SE (n = 3). 1st cut and 2nd cut means the first and
second cutts, respectively.
Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al. Saudi Journal of Biological Sciences xxx (xxxx) xxx
9
0.140 and 0.172 mL plant
1
in both cuts of the second season) and
the greatest essential oil yield per hectare (6.87 and 7.56 L ha
1
in
the first season and 7.76 and 9.57 L ha
1
in the second season) for
both cuts, respectively were obtained from the combination treat-
ment of 30 cm between plants in row and 75 mM melatonin, the
maximum essential oil content per plant (0.154 mL plant
1
)in
the second cut of the first season was found with the combination
treatment of 45 cm between plants and 75 mM melatonin concen-
tration before planting.
3.8. Essential oil constituents
Figs. 1, 2, 3 and Table 18 shows the variable changes in the rel-
ative percentages of different ingredients of the essential oil dis-
tilled from the herb of lovage. It can be remarked that about 37
compounds were identified from essential oil of herb of the first
and second cuts that represented 99.91–100% of lovage essential
oil. The total oxygenated compounds that were ranged from
66.87 to 82.68% were superior comparing with hydrocarbon
Table 14
The mean values of antioxidant content (mg TE 100 g
1
herb) of lovage plants as affected with planting spacing, melatonin concentrations and their combination treatments in
two cuts of both seasons.
Treatments First season Second season
1
st
cut 2
nd
cut 1
st
cut 2
nd
cut
Planting space between plants in row (cm)
15 53.94 ± 0.82 a*54.49 ± 1.52 a 56.63 ± 1.18 a 55.57 ± 1.49 a
30 48.34 ± 2.27 b 47.83 ± 2.29 b 49.61 ± 1.65 b 48.24 ± 2.62 b
45 40.02 ± 3.00 c 39.96 ± 2.58 c 41.48 ± 3.24 c 40.32 ± 2.48 c
Melatonin concentrations (mM)
Without soaking 45.42 ± 7.61 c 45.37 ± 6.72 b 47.20 ± 7.97 d 45.46 ± 7.12 d
0 46.94 ± 6.31 b 46.36 ± 7.38 b 48. 17 ± 7.29 c 47.30 ± 7.31 c
75 50.11 ± 4.65 a 49.57 ± 5.20 a 51. 66 ± 5.58 a 50.57 ± 5.75 a
100 47.26 ± 5.93 b 48.41 ± 6.27 a 49. 94 ± 5.61 b 48.84 ± 6.50 b
Interaction
T1 53.73 ± 0.83 a 52.97 ± 1.34 a 55.65 ± 0.60 b 54.00 ± 0.40 b
T2 53.88 ± 0.91 a 54.73 ± 0.31 a 56.37 ± 1.72 ab 55.47 ± 0.23 ab
T3 54.32 ± 1.06 a 55.33 ± 0.83 a 57.79 ± 0.51 a 56.95 ± 1.60 a
T4 53.82 ± 0.87 a 54.93 ± 2.27 a 56.72 ± 0.73 ab 55.87 ± 1.67 a
T5 46.26 ± 0.33 c 45.55 ± 1.12 cd 48.55 ± 0.48 d 44.69 ± 1.33 e
T6 47.51 ± 0.89 c 46.44 ± 2.15 c 48.47 ± 0.48 d 47.73 ± 1.66 d
T7 51.86 ± 0.64 b 49.87 ± 1.29 b 52.20 ± 0.52 c 50.93 ± 0.46 c
T8 47.70 ± 0.64 c 49.47 ± 0.46 b 49.23 ± 0.49 d 49.60 ± 0.40 c
T9 36.25 ± 0.48 f 37.59 ± 0.35 f 37.41 ± 0.38 g 37.70 ± 0.31 g
T10 39.42 ± 0.48 e 37.90 ± 0.98 f 39.67 ± 0.40 f 38.71 ± 0.35 g
T11 44.15 ± 0.69 d 43.50 ± 0.66 d 44.98 ± 0.72 e 43.82 ± 0.18 e
T12 40.26 ± 1.02 e 40.84 ± 0.63 e 43.86 ± 0.44 e 41.04 ± 0.30 f
*
Means followed by the same letter are not significantly different by the protected LSD, P 0.05. Data are means value ± SE (n = 3). 1st cut and 2nd cut means the first and
second cutts, respectively.
Table 15
The mean values of essential oil percentage (%) of lovage plants as affected with plant spacing, melatonin concentrations and their combination treatments in two cuts of both
seasons.
Treatments First season Second season
1
st
cut 2
nd
cut 1
st
cut 2
nd
cut
Planting space between plants in row (cm)
15 0.568 ± 0.061 a*0.455 ± 0.064 a 0.569 ± 0.103 a 0.494 ± 0.073 a
30 0.530 ± 0.070 b 0.413 ± 0.043 b 0.558 ± 0.067 a 0.450 ± 0.07 2b
45 0.374 ± 0.042 c 0.322 ± 0.051 c 0.402 ± 0.039 b 0.352 ± 0.05 2c
Melatonin concentrations (mM)
Without soaking 0.425 ± 0.064 c 0.330 ± 0.051 d 0.419 ± 0.042 c 0.341 ± 0.053 d
0 0.489 ± 0.113 b 0.386 ± 0.061 c 0.509 ± 0.097 b 0.430 ± 0.07 4c
75 0.559 ± 0.096 a 0.447 ± 0.049 a 0.597 ± 0.104 a 0.503 ± 0.080 a
100 0.490 ± 0.102 b 0.425 ± 0.090 b 0.515 ± 0.095 b 0.455 ± 0.06 0b
Interaction
T1 0.488 ± 0.007 e 0.372 ± 0.016 cd 0.415 ± 0.006 f 0.401 ± 0.031 d
T2 0.594 ± 0.038 abc 0.441 ± 0.018 b 0.584 ± 0.011 cd 0.484 ± 0.011 bc
T3 0.629 ± 0.020 a 0.495 ± 0.040 a 0.687 ± 0.013 a 0.578 ± 0.063 a
T4 0.563 ± 0.053 bcd 0.513 ± 0.042 a 0.591 ± 0.010 c 0.510 ± 0.017 bc
T5 0.437 ± 0.028 f 0.353 ± 0.015 de 0.467 ± 0.018 e 0.336 ± 0.006 e
T6 0.524 ± 0.034 de 0.409 ± 0.017 bc 0.562 ± 0.022 d 0.474 ± 0.01 1c
T7 0.611 ± 0.040 ab 0.448 ± 0.022 b 0.639 ± 0.036 b 0.517 ± 0.01 3b
T8 0.549 ± 0.013 cd 0.444 ± 0.190 b 0.565 ± 0.023 cd 0.474 ± 0.01 1c
T9 0.349 ± 0.022 g 0.265 ± 0.011 g 0.374 ± 0.015 g 0.284 ± 0.00 7f
T10 0.349 ± 0.022 g 0.309 ± 0.013 f 0.382 ± 0.020 g 0.332 ± 0.006 e
T11 0.437 ± 0.280 f 0.397 ± 0.016 c 0.464 ± 0.018 e 0.413 ± 0.027 d
T12 0.359 ± 0.007 g 0.316 ± 0.016 ef 0.390 ± 0.003 fg 0.379 ± 0.009 d
*
Means followed by the same letter are not significantly different by the protected LSD, P 0.05. Data are means value ± SE (n = 3). 1st cut and 2nd cut means the first and
second cutts, respectively.
Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al. Saudi Journal of Biological Sciences xxx (xxxx) xxx
10
compounds which were ranged from 17.32 to 33.04 under all stud-
ied combination treatments in both cuts. The first major compound
is
a
-terpinyle acetate (62.79:77.72%) followed by b-Phellandrene
(10.24:19.18%). The maximum relative pronounce of
a
-terpinyle
acetate (Fig. 4) were obtained with the combination treatment of
15 cm plant spacing and 0 mM melatonin in the first cut while
the maximum relative pronounce of b-Phellandrene (19.18%) was
recorded with the combination treatment of 30 cm plant spacing
and seeds soaking solution of 75 mM melatonin in the second cut.
3.9. Simple correlation coefficients
Table 19, 20 shows the simple correlation coefficients among
various studied characters of lovage in this study as affected with
plant spacing and seeds soaking combination treatments in the
first and second seasons. The symbol (*) refers to significant corre-
lation, (
**
) means high significant correlation while (
***
) means high
high significant correlation. (–) if it was observed it means negative
correlation and if it is not found means positive correlation. The
Table 16
The mean values of essential oil content (mL plant
1
) of lovage plants as affected with plant spacing, melatonin concentrations and their combination treatments in two cuts of
both seasons. Data are means value ± SE (n = 3).
Treatments First season Second season
1
st
cut 2
nd
cut 1
st
cut 2
nd
cut
Planting space between plants in row (cm)
15 0.044 ± 0.008 c*0.044 ± 0.012 b 0.044 ± 0.011 c 0.058 ± 0.013 c
30 0.101 ± 0.019 a 0.110 ± 0.022 a 0.113 ± 0.020 a 0.142 ± 0.032 a
45 0.089 ± 0.013 b 0.111 ± 0.029 a 0.095 ± 0.017 b 0.125 ± 0.028 b
Melatonin concentrations (mM)
Without soaking 0.063 ± 0.022 c 0.064 ± 0.028 d 0.066 ± 0.027 d 0.073 ± 0.025 d
0 0.075 ± 0.074 b 0.081 ± 0.032 c 0.079 ± 0.029 c 0.106 ± 0.040 c
75 0.095 ± 0.096 a 0.115 ± 0.046 a 0.106 ± 0.037 a 0.134 ± 0.047 a
100 0.078 ± 0.078 b 0.094 ± 0.028 b 0.084 ± 0.033 b 0.120 ± 0.047 b
Interaction
T1 0.035 ± 0.002 e 0.028 ± 0.002 h 0.030 ± 0.004 h 0.040 ± 0.007
T2 0.045 ± 0.005 de 0.039 ± 0.002 gh 0.044 ± 0.002 g 0.057 ± 0.001 h
T3 0.055 ± 0.004 cde 0.054 ± 0.003 fg 0.059 ± 0.004f 0.073 ± 0.005 g
T4 0.042 ± 0.003 e 0.056 ± 0.008 f 0.043 ± 0.003 g 0.064 ± 0.003 gh
T5 0.078 ± 0.005 bcd 0.083 ± 0.007 e 0.088 ± 0.004 de 0.090 ± 0.002 f
T6 0.097 ± 0.018 ab 0.103 ± 0.009 cd 0.108 ± 0.012c 0.149 ± 0.006 cd
T7 0.124 ± 0.003 a 0.136 ± 0.008b 0.140 ± 0.007 a 0.172 ± 0.006 a
T8 0.105 ± 0.006 ab 0.117 ± 0.018 c 0.118 ± 0.001 bc 0.156 ± 0.006 bc
T9 0.078 ± 0.007 bcd 0.079 ± 0.012 e 0.080 ± 0.002 e 0.089 ± 0.008 f
T10 0.082 ± 0.007 bc 0.101 ± 0.006 d 0.086 ± 0.009 de 0.111 ± 0.001 e
T11 0.108 ± 0.009 ab 0.154 ± 0.010 a 0.120 ± 0.007 b 0.158 ± 0.004 b
T12 0.087 ± 0.003 bc 0.110 ± 0.004 cd 0.093 ± 0.002 d 0.140 ± 0.001 d
*
Means followed by the same letter are not significantly different by the protected LSD, P 0.05. Data are means value ± SE (n = 3). 1st cut and 2nd cut means the first and
second cutts, respectively.
Table 17
The mean values of yield of essential oil (l ha
1
) of lovage plants as affected with plant spacing, melatonin concentrations and their combination treatments in two cuts of both
seasons.
Treatments First season Second season
1
st
cut 2
nd
cut 1
st
cut 2
nd
cut
Planting space between plants in row (cm)
15 4.91 ± 0.39 b*4.92 ± 0.58 b 4.87 ± 0.53 b 6.49 ± 0.62 b
30 5.61 ± 0.45 a 7.52 ± 1.12 a 6.30 ± 0.48 a 7.87 ± 0.77 a
45 3.28 ± 0.21 c 4.11 ± 0.45 c 3.51 ± 0.26 c 4.62 ± 0.43 c
Melatonin concentrations (mM)
Without soaking 3.69 ± 0.29 c 3.56 ± 0.37 c 3.70 ± 0.39 c 4.22 ± 0.35 d
0 4.46 ± 0.52 b 6.42 ± 1.56 a 4.69 ± 0.54 b 6.25 ± 0.76 c
75 5.66 ± 0.55 a 6.50 ± 0.39 a 6.26 ± 0.63 a 7.84 ± 0.69 a
100 4.58 ± 0.49 b 5.59 ± 0.56 b 4.92 ± 0.57 b 6.99 ± 0.64 b
Interaction
T1 3.84 ± 0.19 fg 3.11 ± 0.20 fg 3.28 ± 0.46 d 4.40 ± 0.75 gh
T2 4.97 ± 0.61 de 4.36 ± 0.20 e 4.88 ± 0.20 c 6.37 ± 0.10 d
T3 6.12 ± 0.42 b 6.02 ± 0.27 c 6.59 ± 0.46 b 8.08 ± 0.51 b
T4 4.69 ± 0.32 e 6.21 ± 0.85 c 4.74 ± 0.35 c 7.12 ± 0.30 c
T5 4.34 ± 0.32 ef 4.64 ± 0.41 de 4.87 ± 0.21 c 4.97 ± 0.11 fg
T6 5.38 ± 0.98 cd 11.36 ± 1.06 a 6.02 ± 0.67 b 8.25 ± 0.34 b
T7 6.87 ± 0.20 a 7.56 ± 0.44 b 7.76 ± 0.38 a 9.57 ± 0.36 a
T8 5.84 ± 0.35 bc 6.51 ± 0.99 bc 6.56 ± 0.05 b 8.67 ± 0.34 b
T9 2.88 ± 0.25 h 2.94 ± 0.45 g 2.96 ± 0.06 d 3.30 ± 0.29 i
T10 3.03 ± 0.24 h 3.74 ± 0.21 efg 3.18 ± 0.33 d 4.13 ± 0.04 h
T11 3.99 ± 0.32 f 5.69 ± 0.39 cd 4.44 ± 0.25 c 5.86 ± 0.15 de
T12 3.21 ± 0.12 gh 4.06 ± 0.16 ef 3.45 ± 0.06 d 5.19 ± 0.03 ef
*
Means followed by the same letter are not significantly different by the protected LSD, P 0.05. Data are means value ± SE (n = 3). 1st cut and 2nd cut means the first and
second cutts, respectively.
Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al. Saudi Journal of Biological Sciences xxx (xxxx) xxx
11
correlation relationship between plant parameters are in harmony
with Hendawy et al. (2015) on Mintha piperita var. citrata and
Hendawy et al. (2017) Rosmarinus officinalis, respectively as well
as Atteya et al. (2020) on Anthriscus cerefolium.
3.10. Antimicrobial activity evaluation:
Lovage volatile oil (L. officinale) exhibits high antibacterial and
antifungal properties in the range of concentrations 75–100 mgmL
1
of oil concentration (Table 21).
4. Discussion
Due to the shortage of information about the response of lovage
plants to agriculture practices in general. and as a now import
plant under Egyptian environmental conditions this work was
done to obtain the optimum combination treatment of soaking
seeds in melatonin before planting and plant density for sowing
lovage seeds. From the above mentioned results of combination
treatments, the treatment of 15 cm between plants in row plus
75 mM melatonin gave the maximum yield of roots fresh weight,
total phenolic and antioxidant content and essential oil percentage
in both seasons. The treatment of 30 cm between plants in row
plus soaking in 75 mM melatonin solution recorded the maximum
plant height, yield of herb fresh and dry weight per ha
1
, yield of
roots dry weight of the first season, essential oil content per plant
and yield per hectare in both season except plant height in the sec-
ond cut of the second season. The treatment of 30 cm between
plants in row plus soaking seeds in 100 mM melatonin had the
maximum plant height and chlorophyll content in the second cut
of the second season. The treatment of 45 cm between plants in
First cut
Second cut
Fig. 1. The pictogram of GC–MS for 15 cm planting space plus without soaking treatment in both cuts.
Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al. Saudi Journal of Biological Sciences xxx (xxxx) xxx
12
row plus using 75 mM melatonin solution recorded the maximum
herb fresh and dry weight per plant and roots fresh and dry weight
per plant, chlorophyll content in both seasons except the chloro-
phyll content in the second cut of the second season.
Plant density is one of the important management factors that
influences the yield of grown plants (Ren et al., 2017; Liu et al.,
2019; Cong et al., 2019; Tian et al., 2020). The plant density influ-
ences plant growth, radiation interception, water consumption,
disease resistance, weed competition and finally, crop yield
(López-Bellido et al., 2005). With decreasing the plant densities
the competition on natural resources such as competition for light
aboveground and competition on more than 20 nutrient elements
and water of irrigation belowground will be also decreased so the
health and productivity of individual plants will become more bet-
ter (Casper and Jackson, 1997; Tian et al., 2020). Therefore, in this
study the planting space of 45 cm between plants in the same row
gave the maximum herb and roots fresh and dry weight per plant
and chlorophyll content, While recorded the minimum essential oil
percentage, phenolic and antioxidant content compared with the
other two less planting space. Moreover, with decreasing the plant-
ing space between cultivated plants the photosynthetic capacity
was increased by increasing the interception of available solar
radiation (Andrade et al., 2002(, exploit all field area resulting in
maximizing herb and roots yield per hectare due to increasing
the number of harvested plants despite of decreasing the weight
of every plant alone and the secondary metabolites such as essen-
tial oil percentage, phenolic content and antioxidant content
reached its maximum level with the less planting space (15 cm).
From the other hand, the most suitable planting space differs from
plant to the other plant and differs from parameter to the other
one. Therefore, despite recording the maximum yield per hectare
of fresh and dry weight of roots with the planting space of 15 cm
between plants in the same row, the tallest plants, the maximum
yield of herb fresh and dry weight per hectare as well as essential
First cut
Second cut
Fig. 2. The pictogram of GC–MS for 30 cm planting space plus 75 mM melatonin treatment in both cuts.
Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al. Saudi Journal of Biological Sciences xxx (xxxx) xxx
13
oil content per plant and yield per hectare of lovage plants were
found with planting space of 30 cm between plants in the same
row. Moreover, due to the fact of decreasing the available water
of irrigation for every plant with increasing the number of plants
in the same area so the degree of water deficit increase and reach
the maximum level with the most narrow space. Polyphenol syn-
thesis and accumulation in plants are generally stimulated in
response to the resistance of plants to environmental stresses
and particularly in water deficit stress (Nacif de Abreu and
Mazzafera, 2005; Giorgi et al., 2009). These results could be
explained by the fact that water deficit may be associated with
an increase in secondary metabolites such as essential oil percent-
age, phenolic and antioxidant content through the reallocation of
the assimilated carbon as plant growth is progressively reduced.
In this condition melatonin is an efficient growth regulator in
plants (Li et al., 2018; Arnao and Hernández-Ruiz, 2014). Soaking
seeds in melatonin promotes seed emergence. As it increases the
chance of survival by enhancing starch metabolism and energy
supplies in response to emergence process (Zhang et al., 2015;
Liu et al., 2015). After emergence, these applications of melatonin
have an important role in increasing photosynthetic rates by
improving plant antioxidant defense drought (Li et al., 2016). In
addition, the idea of soaking seeds before sowing is aimed at short-
ening the lag phase in emergence and to enhance seedling estab-
lishment, thereby minimizing the risk in the early vegetative
growth (Sabongari and Aliero, 2004) and finally enhance growth
parameter. Each crop cultivar requires a critical soaking condition
for good emergence (Harris et al., 2000). So in this work, despite
Fig. 3. The pictogram of GC–MS for 30 cm planting space plus 100 mM melatonin treatment in both cuts.
Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al. Saudi Journal of Biological Sciences xxx (xxxx) xxx
14
soaking lovage seeds in 100 mM melatonin solution gave the min-
imum number of days to emergence, the soaking in 75 mM mela-
tonin before planting them accelerated emergence and also had
the maximum emergence percentage compared with the other
treatments including the control treatment (without soaking).
These good conditions of emergence had influenced role on lovage
plants growth so, using 75 mM melatonin solution for soaking seeds
of lovage before planting gave the maximum mean values of all
studied parameters of herb, roots and chemical constituents in this
study in both cuts of both seasons, respectively compared with
other treatments except for plant height and chlorophyll content
of the second cut in the second season as the maximum values
of it were found with 100 mM melatonin solution across all studied
planting spaces. Moreover, the improving that was found in this
study as a result of soaked seeds compared with unsoaked seeds
is in harmony with Arshad Ullah et al. (2002) who reported that
pre-sowing treatments of raya seed encourage growth of crop,
reduced days to emergence, increased numbers of branches
plant
1
and plant yield. Mozumder and Hossain (2013) found that
days to first emergence decreased with increasing soaking period
of Eryngium foetidum L. seeds. Sabongari and Aliero (2004)
Table 18
Chemical analysis of lovage herb essential oil as affected with planting spacing and melatonin concentrations combination treatments in the second season.
Essential oil compounds Molecular formula RT Relative percentage of the main constituents
15 cm planting space
plus without soaking
30 cm planting space
plus 75 mM melatonin
30 cm planting space
plus 100 mM melatonin
1st cut 2nd cut 1st cut 2nd cut 1st cut 2nd cut
1
a
-Thujene C
10
H
16
4.49 0.00 0.11 0.12 0.16 0.00 0.13
2
a
-Pinene (-)- C
10
H
16
4.69 0.57 0.74 1.11 1.70 1.13 1.09
3 Camphene C
10
H
16
5.13 0.18 0.20 0.37 0.42 0.37 0.27
4 Sabinene C
10
H
16
5.71 0.53 0.68 0.93 0.88 0.67 0.76
52-
a
-Pinene C
10
H
16
5.87 0.27 0.31 0.46 0.46 0.69 0.30
6
a
-Myrcene C
10
H
16
6.13 0.44 1.71 1.22 1.64 1.25 1.82
7
a
-Phellandrene C
10
H
16
6.71 0.22 0.27 0.29 0.46 0.30 0.29
8
a
-Terpinene C
10
H
16
7.03 0.16 0.20 0.21 0.23 0.17 0.21
9 o-Cymene C
10
H
14
7.34 0.00 0.30 0.16 0.18 0.17 0.16
10 (+)-carvene C
10
H
16
7.43 3.33 4.44 4.81 5.45 4.37 5.09
11 b-Phellandrene C
10
H
16
7.52 10.24 10.69 13.28 19.18 15.88 13.67
12
a
-Ocimene C
10
H
16
7.62 0.00 1.29 4.78 0.00 2.60 2.85
13 3-Carene C
10
H
16
7.99 0.00 0.00 0.53 0.00 0.42 0.07
14 ç-Terpinene C
10
H
16
8.43 0.32 0.83 0.29 0.48 0.29 0.36
15
a
-Pinene oxide C
10
H
16
O 9.00 0.00 0.00 0.00 0.00 0.00 0.07
16
a
-Terpinolene C
10
H
16
9.38 0.66 1.06 1.18 1.71 0.91 1.18
17 cis-Ocimene C
10
H
16
12.25 0.00 0.00 0.00 0.00 0.00 0.07
18 l-Menthone C
10
H
18
O 12.39 1.37 0.00 0.00 0.00 0.00 0.00
19 Menthol C
10
H
20
O 13.29 0.54 0.00 0.00 0.00 0.00 0.00
20 l-4-Terpineol C
10
H
18
O 13.35 0.00 0.15 0.27 0.08 0.16 0.21
21 Cryptone (CAS) C
9
H
14
O 13.84 0.00 0.00 0.09 0.00 0.00 0.00
22
a
-Terpineol C
10
H
18
O 14.03 1.37 1.87 2.10 0.96 2.24 2.34
23 Bornyl acetate C
12
H
20
O
2
17.55 0.52 0.34 0.85 0.48 0.70 0.49
24 l-Menthyl acetate C
12
H2
2
O
2
17.80 0.27 0.00 0.00 0.00 0.00 0.00
25
a
-terpinyle acetate C
12
H
20
O
2
20.24 77.72 71.90 63.85 62.79 65.09 65.69
26 Isobornyl propionate C
13
H
22
O
2
21.36 0.09 0.00 0.00 0.08 0.00 0.00
27 Geranyl acetate C
12
H
20
O
2
21.66 0.00 0.38 1.06 0.35 0.48 0.58
28
a
-elemene C
15
H
24
21.77 0.10 0.00 0.00 0.00 0.00 0.00
29 Caryophyllene C
15
H
24
22.96 0.21 0.00 0.00 0.00 0.00 0.00
30 Germacrene-D C
15
H
24
25.51 0.00 0.10 0.00 0.09 0.14 0.09
31
a
-Selinene C
15
H
24
25.87 0.09 0.07 0.00 0.00 0.00 0.00
32 Gynoval C
15
H
26
O
2
26.59 0.27 0.21 0.00 0.26 0.36 0.00
33
a
-Eudesmol (CAS) C1
5
H
26
O 32.45 0.00 0.00 0.00 0.00 0.11 0.00
34 Globulol C
15
H
26
O 32.80 0.00 0.00 0.00 0.00 0.14 0.00
35 Butylidene phthalide C
12
H
12
O
2
33.56 0.00 0.12 0.00 0.08 0.00 0.08
36 Ligustilide C
12
H
14
O
2
35.45 0.53 2.03 1.61 1.79 1.35 2.13
37 Leinoleic acid C
18
H
32
O
2
48.62 0.00 0.00 0.43 0.00 0.00 0.00
Total identified compounds 100.00 100.00 100.00 99.91 99.99 100.00
Total hydrocarbon compounds 17.32 23.00 29.74 33.04 29.36 28.41
Total Oxygenated compounds 82.68 77.00 70.26 66.87 70.63 71.59
1st cut and 2nd cut means the first and second cutts, respectively.
Fig. 4. Structure of
a
-terpinyle acetate present in herb essential oil of lovage by
using GC–MS analysis.
Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al. Saudi Journal of Biological Sciences xxx (xxxx) xxx
15
observed that soaking the seeds of Lycopersicum esculentum Mill in
water before sowing them in the field improved emergence and
growth compared with control seeds. Therefore, under normal con-
dition (wide planting spacing) and water-stress conditions (narrow
planting spacing) soaking seeds in melatonin solution was able to
improve emergence and growth as well as all studied parameters
and chemical composition. It was in harmony with all of Jiahao
et al. (2019), Zhang et al. (2014), Li et al. (2016) and Chen et al.
(2020) who found that exogenous melatonin application improved
emergence and growth of seeds and plants under normal and abi-
otic stresses condition.
The major components of essential oil of lovage herb in the first
and second cuts were b-Terpinyl acetate and b-Phellandrene as
agree with Samiee et al. (2006) and Mohamadi et al. (2016).It
can be mentioned that the antimicrobial properties of the lovage
essential oil are due to the high content of b-phellandrene,
a
-
terpinyl acetate and Ligustilide. The mentioned compounds exhibit
pronounced antimicrobial properties through mechanisms that
Table 19
Simple correlation coefficients among various studied characters of lovage in this study as affected with plant spacing and melatonin concentration combination treatments in the
first season.
123456789101112131415
1 1.000
2 0.329 1.000
3 0.634
***
0.387* 1.000
4 0.280 0.979
***
0.324 1.000
5 0.505
**
0.580
***
0.843
***
0.612
***
1.000
6 0.230 0.980
***
0.354* 0.953
***
0.542
***
1.000
70.479
**
0.205 0.249 0.170 0.180 0.308 1.000
8 0.728
***
0.394* 0.604
***
0.385* 0.573
***
0.290 0.758
***
1.000
9 0.320 0.036 0.280 0.037 0.091 0.024 0.378 0.190 1.000
10 0.629
***
0.384* 0.506
**
0.355* 0.447
**
0.288 0.623
***
0.899
***
0.179 1.000
11 0.024 0.823
***
0.124 0.801
***
0.091 0.848
***
0.509
**
0.013 0.013 0.099 1.000
12 0.047 0.829
***
0.099 0.818
***
0.133 0.844
***
0.480
**
0.027 0.079 0.094 0.968
***
1.000
13 0.329 0.534
***
0.419* 0.527
***
0.205 0.580
***
0.664
***
0.369* 0.068 0.250 0.867
***
0.846
***
1.000
14 0.423* 0.878
***
0.606
***
0.898
***
0.823
***
0.846
***
0.092 0.576
***
0.095 0.462
**
0.500
**
0.517
**
0.136 1.000
15 0.348* 0.078 0.638
***
0.054 0.539
***
0.119 0.348* 0.423* 0.073 0.262 0.273 0.235 0.504
**
0.379* 1.000
1. Plant height 2. Herb fresh weight per plant 3. Herb fresh weight per hectare 4. Herb dry weight per plant 5. Herb dry weight per hectare 6. Roots fresh weight per plant 7.
Roots fresh weight per hectare 8. Roots dry weight per plant 9. Roots dry weight per hectare 10. Chlorophyll content 11. Total phenolic 12. Antioxidant content 13. Essential
oil percentage 14. Essential oil content per plant 15. Essential oil content per hectare *. Significant
**
. High significant.
-
: Negative relationship.
Table 20
Simple correlation coefficients among various studied characters of lovage in this study as affected with plant spacing and seeds soaking combination treatments in the second
season.
123456789101112131415
1 1.000
2 0.225 1.000
3 0.487
**
0.412* 1.000
4 0.249 0.576
***
0.399* 1.000
5 0.544
***
0.394* 0.933
***
0.403* 1.000
6 0.136 0.573
***
0.302 0.982
***
0.272 1.000
70.662
***
0.105 0.147 0.207 0.265 0.335* 1.000
8 0.774
***
0.250 0.363* 0.428
**
0.461
**
0.320 0.717
***
1.000
9 0.027 0.032 0.053 0.001 0.087 0.036 0.451
**
0.265 1.000
10 0.448
**
0.218 0.316 0.343* 0.488
**
0.238 0.395* 0.693
***
0.084 1.000
11 0.138 0.357* 0.102 797
***
0.129 0.872
***
0.561
***
0.047 0.186 0.022 1.000
12 0.164 0.394* 0.132 0.787
***
0.161 0.866
***
0.560
***
0.037 0.171 0.005 0.978
***
1.000
13 0.557
***
0.160 0.450
**
0.419* 0.457
**
0.536
***
0.725
***
0.379* 0.068 0.290 0.816
***
0.831
***
1.000
14 0.513
**
0.589
***
0.710
***
0.875
***
0.725
***
0.790
***
0.132 0.615
***
0.082 0.489
**
0.425
**
0.406* 0.045 1.000
15 0.649
***
0.143 0.805
***
0.014 0.847
***
0.159 0.574
***
0.488
**
0.048 0.463
**
0.553
***
0.583
***
0.856
***
0.450
**
1.000
1. Plant height 2. Herb fresh weight per plant 3. Herb fresh weight per hectare 4. Herb dry weight per plant 5. Herb dry weight per hectare 6. Roots fresh weight per plant 7.
Roots fresh weight per hectare 8. Roots dry weight per plant 9. Roots dry weight per hectare 10. Chlorophyll content 11. Total phenolic 12. Antioxidant content 13. Essential
oil percentage 14. Essential oil content per plant 15. Essential oil content per hectare *. Significant
**
. High significant.
-
: Negative relationship.
Table 21
The antimicrobial activity (MBC, MFC) * of the oil extracted from the lovage plants.
Examined Bacteria and Fungi Oil concentration mgmL
1
0255075100
Ralstonia solanacearum
(4.8 10
8
CFU mL
1
)–––++
Pectobacterium Carotovorum subsp atrosepticum
(4.8 10
8
CFU mL
1
)–––++
Pectobacterium Carotovorum subsp Carotovorum
(4.8 10
8
CFU mL
1
)–––++
Fusarium roseum (3.0 10
7
CFU mL
1
)–––++
*MBC - minimal bactericidal concentration; MFC - minimal fungicidal concentration.
Amira K.G. Atteya, A.N. Albalawi, H.M. Bayomy et al. Saudi Journal of Biological Sciences xxx (xxxx) xxx
16
include breaking of the cell wall and cytoplasmic membrane,
reduction of the cytoplasm around the nucleus, disturbance of
the lipid fraction of the plasma membrane resulting in the alter-
ation of its permeability and the leakage of the intracellular con-
tent (Li et al., 2014; Trombetta et al., 2005).
5. Conclusion
From this work, it appeared that the growth of the lovage plant
significantly affected by different treatments in this study. More-
over, measurements comprising of herb fresh and dry weight as
well as essential oil yield per hectare showed that the combination
treatment of 30 cm between putting plants in row plus soaking
seeds in 75 mM melatonin solution was able to achieve the maxi-
mum values of these parameters. While the combination treat-
ment of 15 cm between plants in row plus seeds soaking in
75 mM melatonin solution is recommended for getting the maxi-
mum yield of roots fresh and dry weight per hectare and maximum
polyphenols, antioxidant content per herb in both cuts of both
studied season. The maximum relative pronounce of
a
-terpinyle
acetate were obtained with the combination treatment of 15 cm
planting space and 0 mM melatonin. While the maximum relative
pronounce of b-Phellandrene was recorded with the combination
treatment of 30 cm plant spacing and seeds soaking solution of
75 mM melatonin
Funding
This research received no external funding.
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: The data presented in this study
are available within the article.
Declaration of Competing Interest
The authors declare that they have no known competing finan-
cial interests or personal relationships that could have appeared
to influence the work reported in this paper.
Acknowledgements
This work did not have any funding support.
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