1.Introduction
Parsley
(Petroselinum crispum) is a herbaceous plant from the Apiaceae family,
originally found in Southwestern Europe and Western Asia. It has been cherished
since ancient Greek times for its culinary and medicinal uses (Zohary et al.,
2012; Dalby, 2003; Riddle, 1985). Known worldwide for its versatility, parsley
is used in cooking, traditional medicine, and as an aromatic herb. In
Arabic-speaking regions, it’s referred to as baqdwnis or maqdwnis.
Fresh parsley leaves are often used as a garnish, while dried leaves, known as
parsley flakes, are popular in processed foods like soups and sausages.
Medicinally, parsley is valued for its carminative, diuretic, hypotensive,
stomachic, nervine, and emmenagogue properties, with its distinctive flavor and
aroma coming from its essential oil.
In
Sudan, parsley has grown from a minor crop to a key ingredient in local cuisine
and traditional medicine (Ahmed et al., 2023; Osman et al., 2021). It’s
primarily cultivated as a winter crop and adapts well to different soil types,
though it prefers light, well-drained soils. The harvesting period lasts about
five months, beginning two months after planting and continuing monthly. Local
varieties, likely introduced through Egypt, are managed by Sudanese farmers
without formal documentation, which affects their conservation. In Central and
Northern Sudan, two native cultivars perform well in various agro-climatic
conditions. However, research on these varieties’ distinctiveness and agronomic
traits is sparse, despite their cultivation in arid and semi-arid regions.
This
study seeks to fill this research gap by examining the growth, yield, and
essential oil properties of these local parsley lines. Insights gained will
help improve cultivation practices, boost productivity, and support effective
breeding and reintroduction efforts.
2. Materials and Methods
2.1. Materials
Two local lines of parsley
(Petroselinum crispum) namely Dongla and Mesdani, were collected from the seed
market of North Sudan, Dongola City and central Sudan, Gezira State. The
research was conducted at the Medicinal and Aromatic Plants Research Institute
(MAPRI) in Shambat locality, Khartoum State, Sudan at (15°40’ N, 32°32’ E) and 383.54
meters above sea level. Seed sowing took place at the winter season, starting
on November 15, 2017. Shambat soil is classified as belonging to the central clay
plain of Sudan and identity as vertisols. This region is shaped by the
deposition of alluvium from the Nile, mainly basaltic in origin. It exhibits a
moderate level of saline and sodic subsoil. The soil composition is
consistently clayed across the experimental farm (Ali et al. 2016). Shambat has
a subtropical desert climate. The locality’s yearly temperature is 31.98ºC
(89.56ºF) and it is 1.94% higher than Sudan’s averages with winter temperatures
ranging from 21-34 ºC. Precipitation is irregular and generally scarce, with
the majority occurring from June to September. It typically receives about
14.28 millimetres (0.56 inches) of precipitation and has 31.03 rainy days (8.5%
of the time) annually. The main metrological data including, minimum, maximum,
and ambient temperature, during the experiment, is presented in Table 1.
2.2 Methods
Experimental design
A four-replication factorial
experiment in a completely randomized block design (CRBD) with two factors,
varieties (A) and frequency of Harvest (B), was examined. Variety had two
levels (Dongola and Madani local Line), while the frequency of Harvest had two
levels (first and second cut). Each plot consisted of four rows, each three
meters long, with seeds sown in paired rows at an inter-row spacing of 15 cm. The
main aims of this experiment were to systematically characterize and assess
existing parsley local lines of Sudan for DUS (Distinctness, Uniformity,
Stability), growth, yield performance and essential oil profile. The research
is also aimed at measuring the resilience of the two local lines as multi-cut
crops through measuring variety harvest frequency interaction. Planting took
place directly in the field on November 15, 2017. Plants were consistently
irrigated based on meteorological conditions, while manual weeding was employed
to manage weed growth. Monitoring of the crop encompassed parameters such as
development, the yield of fresh herbage, and phytosanitary status. Harvesting
was initiated 70-90 days post-planting, with several harvests conducted. After
harvesting, the leaves underwent trimming, washing, bundling, and preservation
at -5°C until essential oil extraction.
Morphological Traits
(Variety descriptors)
Morphological traits were assessed using visual
descriptors by examining a total of sixty plants for each local line. Fifteen
plants were selected randomly from each replication. Plants were hand-harvested
at marketable foliage size by cutting at a height of five cm above the ground.
Two harvests were taken at suitable intervals (70-140 days) from seeding. Fifteen
Variety descriptors were visually scored according to the UPOV code: PETRO_CRI Petroselinum crispum (Mill.)
Nyman ex A.W. Hill, issued by the International Union for the Protection of New
Varieties of Plants (UPOV, 2005). The descriptors include plant height, plant width, the
density of foliage, number of leaves, leaf attitude, leaf
blade curling, leaf blade length, leaf blade width, leaf blade ratio length/width, leaf blade
intensity of green colour, leaflet shape, leaflet
shape, distance between 1st and 2nd pair of leaflets, undulation
of leaflet margin, petiole length and petiole thickness.
The results are presented in Table 2.
Growth parameters
Morphological traits were measured or counted on the
average of 10 plants selected randomly from each plot. Morphological traits include
plant height, number of branches, canopy diameter, leaf blade length,
leaf blade width, the distance between the first and second pair of Leaflets,
petiole length, petiole thickness, number of leaves per plant, fresh weight, and
dry weight. The data on morphological traits are presented in Table 3.
Biomass Yield
Biomass yield was estimated by
harvesting 1m² from the central area of each plot. The yield was recorded in kilograms
per square meter (kg/m²). The biomass yield and fresh and
dry weight are presented in Table 4.
Essential oil yield and
chemical compositions
Isolation of Essential Oils:
One hundred grams of fresh leaves
were washed, dried, and finely ground for optimal extraction. The sample
underwent hydro-distillation for three hours using Clevenger-type equipment. Distillate was collected, and the essential
oil was extracted using dichloromethane dried over anhydrous sodium sulfate,
and stored at 4°C in airtight amber vials
until subjected to GC and GC-MS. Essential oil isolation was carried out as
proposed by Huie (2002).
GC and GC-MS Analysis
The GC-MS analysis was conducted as per the methods and
procedures described in Adams (2007). Essential oil analyses were carried out at the Department of
Medicinal and Aromatic Plants Research, National Research Center, Sudan. 1-μL
injection sample was analysed using a Gas Chromatograph Mass Spectrometer
manufactured by Shimadzu Company’s model GC/MS-QP2010-Ultra, serial number
020525101565SA. A capillary column (Rtx-5 ms-30 m × 0.25 mm × 0.25 µm) was
used. The sample was injected in split mode, with the instrument operating in EI mode at
70 eV. Helium was used as the carrier gas at a flow rate of 1.2 mL/min. The
temperature program started at 50°C and was held for 3 minutes, with a rate of
4°C/min reaching 280°C and held for 10 minutes. The injection port temperature
was 300°C, the ion source temperature was 200°C, and the interface temperature
was 250°C. The sample was analyzed in scan mode in the range of 40–500 m/z. The
identification of components in the sample was done by comparing their
retention index and mass fragmentation patterns with those available in the
National Institute of Standards and Technology (NIST) library. Major essential oil
profiles are presented in Table 5.
Statistical analysis
A conventional factorial RCBD analysis was used to test
the significance of differences between different genotypes. Data were analyzed
using the MSTAT-C program, applying conventional ANOVA analysis to test for
significant differences between varieties, harvest frequency and their
interaction. The critical difference (C.D) at the 5% probability level was used
to determine statistical significance as per the method suggested in Sokal and Rohlf, 1995.
3. Results and Discussion
The climatic conditions during the experiment,
as shown in Table 1, indicated a range of temperatures, humidity, and rainfall
throughout the study period. The average temperatures varied from 22.5°C in
January to 31.8°C. The average temperature across the growing season is 28.35 oC.
Maximum temperatures peaked at 38.8°C in April, while minimum temperatures
dropped to 15.9°C in January. Relative humidity decreased from 28.6% in
December to 8.7% in April, and no rainfall was recorded throughout the
experiment. Several studies demonstrate the significant role of temperature
levels in influencing the physiological processes and fresh mass of parsley. A
primary factor affecting plant growth development is temperature, as modulated
by other factors including day length and vernalization (Hodges, 1991). The
quantitative effects of temperature on plant growth have been studied
extensively using the concept of thermal time (Jamieson et al., 1995; Walters
and Lopez, 2021). An optimum temperature for parsley 'Giant of Italy'
fresh mass is estimated at around 28°C when grown hydroponically under control
conditions, (Walters and Lopez, 2021; Frąszczak and Knaflewski,
2009). The climatic data during this experiment align with recommendations
from previous studies and also support local farmers' preference to cultivate
parsley during this period in Sudan, highlighting the crucial role of
environmental factors in parsley cultivation.
To better understand the varietal differences
between these two local lines, the findings on morphological traits are
presented in Table 2. The morphological trait score adheres to the UPOV code
(International Union for the Protection of New Varieties of Plants, 2005). Dongola
local Line and Madani's local lines exhibit similar traits for leaf attitude
(erect), leaflet shape (medium triangular), leaf blade ratio length/width
(medium), and absence of leaf blade curling. However, all other remaining
characteristics set them apart. The differences observed between these two
lines are sufficiently consistent and distinct from each other. Both lines are
uniform within themselves and can be considered stable without the need for
more than one growing cycle. The geographic isolation between these two lines
and the absence of newly introduced lines have contributed over the years to
stabilize both lines. The absence of leaf blade curling categorizes both lines
under the flat-leaf form of parsley (Petroselinum crispum var.
neapolitanum), also known as Italian parsley, with broad flat leaves and
stronger flavour compared to curly-leaf parsley (Grin 2008). Such kind of
descriptors are useful to identify morphological markers related to aroma and
plant size/yield for productivity, (Boutsika et al., 2021).
Leaf parsley is a great candidate for multi-cut
harvest every 60-70 days from the same plant throughout its growing season. The
reaping and regrowth system is not only feasible for the production of leafy
salad vegetables but has also been used in tea and grass production (Belesky
and Fedder,1995; Fisher and Dowdeswell, 1995; Murtagh and Smith, 1996; Bore et
al., 2003; Nayak and
Maji 2018) .this system is
also useful in other species such as leafy vegetables. The system can be applied to other species such
as parsley. Hence it is essential to measure the growth rate and the total
biomass produced after each cut. This can help determine the plant's ability to
regenerate, recover, and maintain productivity. This can be influenced by the
plant's genetic makeup and intervarietal differences as well as environmental
conditions, and management practices. The two lines were tested and the
findings were presented as per Tables 3 and 4 for growth and biomass yield
parameters.
The Medani
local Line showed significantly higher values for growth parameters at a 5%
level of confidence for leaf blade length (14.542 cm), leaf blade width (19.329
cm), petiole thickness (12.51 mm), and plant height (39.450 cm) compared to Dongola
Local Line. Harvest frequency also affected the growth parameters, with the
first cut showing higher values for most traits except canopy diameter. The
first cut recorded higher values for LBL (15.98 cm), LBW (19.46 cm), leaf blade
distance between 1st and 2nd pair of leaflets (7.612 cm), petiole length (12.51
cm), plant height (45.68), and number of leaves (21.35). The second cut
recorded the highest canopy diameter of (20.61 cm). In multicut cropping
systems, repeated harvesting often increases canopy diameter due to stimulated
branching and leaf production. Regular harvesting can make parsley and similar
plants develop a bushier canopy as apical dominance is removed, encouraging
lateral growth. Similar phenomena were observed in various herbaceous plants
and leafy vegetables, suggesting similar outcomes for parsley (Sanderson et
al., 1995; Fu, J.
2008). An interaction effect was observed between variety and
harvest frequency for petiole thickness and number of leaves at a 5% confidence
level. Dongola local line interacts significantly well showing a higher number
of leaves per plant which indicates a better ability to regenerate, recover,
and maintain productivity.
Table 4.
presents the biomass yield, dry weight, and oil content of the parsley local
line. Dongola local line produced a higher biomass yield of (5.849 kg/m²)
compared to Madani (4.697 kg/m²), although the difference was not statistically
significant. Dongola local Line recorded significantly higher dry weight (2.284
kg/m²) while Medani local line was significantly superior in terms of oil
content (7.88 g/150g and 5.6%). The first cut showed significantly higher
biomass yield and oil content compared to the second cut. Interaction effects
between variety and harvest frequency were not significant for the parameters measured.
The varietal differences observed in this study are consistent with previous
research on parsley genetics and breeding (Fusain, et al. (2016); Al-Attar et al. (2021) and Salehi et
al. (2022). Abed and Shebl (2016) reported similar differences in biomass yield
between the first and second cut in spinach. The study suggests that the
combined harvesting of spinach in the two cuttings can maximize the total fresh
yield, even though the second cutting may produce a smaller yield than the
first cutting.
Comparison
of Essential Oil Components
The Gas Chromatography/Mass
Spectrometry (GC/MS) analysis revealed distinct differences in the essential
oil composition of both lines. The major components identified in both lines included
Alpha-Pinene, Beta-Pinene, Beta-Phellandrene, Alpha-Terpinolene, Myristicin,
Elemicin, and Apiole. The relative concentrations of major essential oil
components in the fresh leaves of the two parsley local lines are presented in
Table 5.
Medani
local line exhibited higher concentrations of Alpha-pinene (8.37%), Beta-pinene
(8.09%), and Alpha-phellandrene (2.05%). Interestingly, Beta-phellandrene was
absent in Madeni but constituted a significant portion of Dongola’s essential
oil profile at (24.67%). This compound is valuable in perfumes, and various
medicinal and cosmetic products (Bentley et al., 2013). A similar unique appearance
for apiole at (0.41%) in the essential oil profile of Dongola local line. Both
varieties contained Myristicin, but Dongola had a higher concentration of
(23.81%) compared to Madeni local line (9.89%). The concentration of Elemicin was
low in both varieties and slightly higher in Dongola recording an amount of (0.77%)
and (0.53%) respectively. Dongla recorded 1,3,8-p-Menthatriene content of
(0.77%) while Madeni local line recorded (0.53%) of 1,3,8-p-Menthatriene. This
essential oil contributes along with others to the aroma of parsley and has
antioxidant properties.
Essential oil composition plays a
crucial role in determining the quality and commercial value of parsley. The
variation in essential oil components between the two lines is consistent with
findings from a study by Boutsika, et
al. 2021 and Fusani, et al. 2016, which
highlighted significant differences in essential oil profiles among parsley
cultivars due to genetic diversity and environmental factors. The significant
difference observed in the concentration of Myristicin between the Medani and Dongola
local Lines of parsley is noteworthy. Myristicin is a naturally occurring
compound found in several essential oils and has been extensively studied for
its bioactive properties, including antifungal, antibacterial, and anticancer
activities. Previous studies indicate that Myristicin, Apiole, and Beta-phellandrene
are commonly found in significant amounts in parsley essential oils. The
results from the Dongola line align with these findings, particularly in its
high Myristicin and Beta-phellandrene content, (Matasyoh et al., 2009; Fusani, et al.
2017; Salehi et al., 2019; Salas
Oropeza et al., 2021; Sany, et al. 2022 ).
The higher concentration of Myristicin in the Dongola local line suggests that
the Dongola local line may have greater potential for use in medicinal
applications where Myristicin is a key active component (Seneme, et al. 2021;
Zheng et al.1992). Alpha-pinene and Beta-Pinene are known for their
anti-inflammatory and bronchodilator effects, making them valuable in the
formulation of therapeutic agents for respiratory conditions ( Salehi et al.,
2019; Salas-Oropeza et al. 2021). Both
local Lines have sufficient concentrations of Alpha-pinene and Beta-Pinene in
their profile, making them equally effective in such therapeutic applications. Beta
Phellandrene and Alpha -Terpinolene have been reported to possess antimicrobial
properties, with potential applications in food preservation and
pharmaceuticals (Vokk, et al. 2011;
Pazyar, et al.2013; Cheng et al. 2017; Salas-Oropeza et al. 2021; Radice et.
al. 2022;). The comparable levels of these compounds in both lines of parsley
suggest that either variety could be used interchangeably for their
antimicrobial benefits. Elemicin and Apiole, though present in lower
concentrations, have also been studied for their biological activities.
Elemicin has been identified as a potential anxiolytic agent (Barman, et al.
2023; Fernandes et al. 2024).), while Apiole is known for its antipyretic,
insecticidal, diuretic, and antitumor effects (Punoševac et al.202). The
consistent presence of these compounds across both local lines further
highlights the therapeutic versatility of parsley essential oils. The absence of Beta-phellandrene in Madeni and its
significant presence in Dongola, as well as the exclusive presence of Alpha-terpinolene
in the Madeni local line, highlights the genetic diversity between the two
lines. Alpha-terpinolene has antioxidant activities, sedative effects,
and antimicrobial effects which make the Madeni local line a great candidate
for this component (Pazyar, et al.2013). The
differences in the essential oil composition of the parsley cultivars are
mainly due to genetic variation. Previous research has also observed such
variability across different parsley varieties and environmental conditions (Fusani, et al.
2017; Bentley et al., 2013). However, other
factors, such as the plant's age, the specific part of the plant studied, its
management, and the environmental conditions in which the plant is grown, can
also affect the percentage of essential oils (Said-Al Ahl, et al. 2016).
4. Conclusions and Recommendations
The study demonstrated significant
variations between the Medani local line and the Dongola local line in terms of
growth parameters, morphological traits, biomass yield, and essential oil
composition. Medani local Line was superior in terms of growth parameters and
essential oil content, while the Dongola local Line had higher biomass yield
and specific essential oil components like Beta-phellandrene. The harvest frequency
also played a critical role, with the first cut generally producing better
results across most parameters. The varietal differences and their response to
harvest frequency provide valuable insights for farmers and breeders in
selecting appropriate varieties and optimizing harvest schedules to maximize yield
and oil content. Overall, the findings
of this study contribute to growing knowledge on the diversity that exists in
local lines of Sudan, the chemical diversity and the potential use of parsley
essential oils. The significant difference in Myristicin concentration warrants
further investigation into the genetic and environmental factors influencing
its biosynthesis in parsley under Sudan conditions. With the growing importance
of the crop in Sudan, we highly recommend the introduction of new lines to
increase the gene pool for any future breeding program.
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This study explores the growth,
yield, and essential oil properties of two Sudanese parsley lines, Dongola and
Madani, focusing on their agronomic and medicinal potential. Parsley (Petroselinum
crispum) is a key culinary herb with significant benefits in regions like
Sudan, where climate conditions are crucial for its cultivation success.
Conducted at the Medicinal and Aromatic Plants Research Institute (MAPRI) in
Shambat, Khartoum State, Sudan, this winter-season experiment commenced on
November 15, 2017. Using a factorial experiment within a completely randomized
block design (CRBD), we examined two factors: variety (A) and cutting frequency
(B). The objectives were to characterize and assess these local parsley lines
for distinctness, uniformity, stability (DUS), growth performance, yield, and
essential oil profile. Additionally, the study evaluated the resilience of the
lines under multi-cut conditions by analyzing interactions between variety and
harvest frequency. Visual descriptors for DUS adhered to the UPOV code (2005).
Both lines exhibited similar leaf characteristics but differed in other traits.
The Medani line showed superior growth metrics, including leaf blade length
(14.54 cm), leaf blade width (19.33 cm), petiole thickness (12.51 mm), and
plant height (39.45 cm). In contrast, Dongola had a higher dry weight (2.284
kg/m²), while Medani excelled in oil content (7.88 g/150g, 5.6%). The first cut
produced higher biomass and oil content. GC/MS analysis revealed distinct
essential oil profiles, with Medani having higher Alpha-Pinene and Beta-Pinene,
while Dongola had higher Myristicin levels, indicating its potential for
medicinal applications.