agriculture
Article
Anatomical Assessment of Skin Separation in Date Palm
(Phoenix dactylifera L. var. Mejhoul) Fruit during Maturation
and Ripening Stages
Nihad Alsmairat, Yahia Othman * , Jamal Ayad
Citation: Alsmairat, N.; Othman, Y.;
Ayad, J.; Al-Ajlouni, M.; Sawwan, J.;
El-Assi, N. Anatomical Assessment
of Skin Separation in Date Palm
(Phoenix dactylifera L. var. Mejhoul)
Fruit during Maturation and
Ripening Stages. Agriculture 2023, 13,
38. https://doi.org/10.3390/
agriculture13010038
Academic Editor: Luca
Corelli-Grappadelli
Received: 28 November 2022
Revised: 18 December 2022
Accepted: 20 December 2022
Published: 22 December 2022
Copyright: © 2022 by the authors.
Licensee MDPI, Basel, Switzerland.
This article is an open access article
distributed under the terms and
conditions of the Creative Commons
Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
4.0/).
Agriculture 2023, 13, 38. https://doi.org/10.3390/agriculture13010038
, Malik Al-Ajlouni , Jamal Sawwan and Najib El-Assi
Department of Horticulture and Crop Science, School of Agriculture, The University of Jordan,
Amman 11942, Jordan
* Correspondence: ya.othman@ju.edu.jo
Abstract: Skin separation (puffiness) is a critical physiological disorder that significantly reduces
the date palm (Phoenix dactylifera L.) fruit’s visual appearance and quality. The objectives of this
study were to analyze fruit anatomy in skin-separated and normal date palm (var. Mejhoul) fruit
and to assess the microclimatic conditions (temperature and relative humidity) during the fruit
developmental stages and their association with skin separation. Fruit anatomy analysis was carried
at different growth stages (Kimri-Khalal, Khalal, Rutab and Tamr) for two growing seasons. In
addition, microclimatic conditions (specifically, temperature, and relative humidity) as well as soil
moisture content were assessed across the study period 2020 and 2021. The anatomical analysis of
the date palm fruit revealed that the exocarp or skin (cuticle, epidermis, hypodermis) thickness were
quite similar across the developmental stages and over the study period (2020, 2021). Conversely,
significantly more sclereid cells were found in skin-separated fruit (compared to normal fruit) at the
Tamr stage. At that stage (Tamr), the percentage increase in sclereid cell number in skin-separated
fruit ranged from 34–183%, sclereid clusters number 23–92%, cluster area 37–206%, and cluster
perimeter 25–64%, as compared to normal fruit. In addition, sclereid cells at skin-separated areas of
the fruit were clustered together, forming a chain of aggregates underneath the fruit skin, and were
closer (42–50%, than normal) to the cuticle, leading to the partial separation of the exocarp from the
fruit mesocarp. Although the weather data were similar across the growing seasons (temperatures,
33–34 ◦ C; relative humidity, 45–46%), skin separation occurred in 14–15% of total fruit in 2020 and
30–34% in 2021. Therefore, we believe that skin separation disorder is not as heavily weather
dependent as it seems to have been thought. In conclusion, changes in microclimate conditions were
not the conclusive component for inducing the skin separation physiological disorder in date palm
fruit. Further studies are required to identify the main factors that stimulate the development of
sclereid clusters aggregates and the potential of agricultural practices to reduce skin separation.
Keywords: sclereid cells; Kimri; Tamr; fruit anatomy; puffiness
1. Introduction
Date palm (Phoenix dactylifera L.) is considered one of the most important fruit crops
in many regions of the world, especially those with hot and dry climates. Date palms
thrive in hot, arid climates, with the majority of their production taking place in the
Southern Mediterranean regions [1]. In fact, date palm trees are adapted to a wide range
of soil conditions including mineral nutrients, salinity, and water stress [2]. Worldwide,
the total area grown is estimated to be ca 1.3 million ha, and total production is about
13 million tons [3]. In 2017, the total number of date trees cultivated globally was about
100 million, out of which ~90% were grown in the Middle East and North Africa regions [4].
In Jordan, the total cultivated area is about 3244 ha, and total production is estimated to be
30,000 tons [3].
https://www.mdpi.com/journal/agriculture
Agriculture 2023, 13, 38 2 of 11

Dates are an important component of healthy diets in many nations and are recognized
as the main food crop in those regions [2]. In fact, the date palm has played a key role in
food security in the Middle East and North African regions by providing nutritional food
for humans for the last 5000 years [4]. Date carbohydrate (total sugars) content ranges from
44 to 88%, fiber 6.4 to 11.5%, protein 2.3 to 5.6%, fat 0.2 to 0.5%, oil 0.2–0.5%, and there
are 15 types of salts and minerals [1]. Furthermore, the seed (accounting for 5.6–14.2%
of the date weight) contains 7.7 to 9.7% oil, 14 types of fatty acids [1], and 15% of fiber
(mainly water-insoluble mannan fibers). Dates are a rich source of phenolic antioxidants
(1–2%), especially condensed tannin and fiber (6.5 to 11.5%), as reported by Ghnimi et al. [4].
Thus, date fruit and seeds are used in some food products as a rich source of fiber and
antioxidants [4]. The high demand for date palm fruit worldwide due to its potential
source of antioxidants and sugars makes it an ideal crop to help with the development of
low-income countries, where poverty is dominant [5].
The fruit development of date palm as reported by Al-Hajjaj et al. [6] can be classified
into five stages, namely, Hababouk (1–5 weeks from pollination, WAP), Kimri (6–16 WAP),
Khalal (17–20 WAP), Rutub (21–24 WAP), and Tamr (25–27 WAP). The fruit at the Tamr stage
have low moisture content (less than 25%), which is appropriate for long-term storage [2,7].
Date sorting is normally done according to shape, size, maturity, weight, condition of
the flesh (ranges from soft to dry), and skin quality [5,8]. The absence of visual defects,
deformity, skin puffiness, sunburn, insect damage, uniformity of color and size, decay,
fermentation, and mechanical damage are some of the criteria that affect the quality grades
of the date fruit [8]. However, fruit quality at the time of harvest and during storage is
influenced by pre-harvest growth conditions during the growing season [9]. These effects
may alter the fruit skin and cause certain subsurface cellular structural alterations, which
may have an impact on the quality and shelf life of the fruit after harvest [9].
The majority of producing countries have large losses during harvest and posthar-
vest processing and marketing as a result of physical and physiological disorders, espe-
cially those associated with skin appearance [2,7]. In most horticultural crops, including
date palm, the visual appearance of the fruit skin is essential for determining its market
value [10]. Physiological disorders that are associated with skin potentially lower the
visual quality of the fruit and result in possible economic loss to the growers [10]. Skin
separation of ripe date fruit (especially “Mejhoul”) lowers the cultivar market value [11].
The price range for fruit with 30–50% skin separation (puffiness) is about 80% lower than
that with an intact, normal skin (no puffiness) [12]. Skin separation (puffiness) normally
occurs when the date skin is dry, hard and brittle [13]. This physiological disorder develops
during ripening and leads to the separation of the skin from the flesh [13]. A few research
studies have been carried out to understand and reduce skin separation [11,14]. How-
ever, the main causes of skin separation in the date palm fruit is not yet fully understood.
Kader and Hussein [13] suggested that high relative humidity and temperature at early
stages of ripening might lead to skin separation in dates. Al-Hajjaj and Ayad [11] found
that boron foliar application at the rate of 1600 mg L−1 significantly increased date yield
(67.7 vs. 53.4 kg tree− 1 ) and fruit size (50 vs. 45 mm) but did not reduce skin separation.
The deficit irrigation technique at the fruit ripening stage stimulated the ripening of the
date palm fruit (while maintain yield) but had limited effect (not significant improvement)
on skin separation [6]. Considering the high economic value of the date palm fruit and the
potential losses associated with skin separation, as well as the uncertainty of identifying
the main causes of this physiological disorder, the study of date palm fruit anatomy and
microclimate is critical to progress the research efforts associated with skin separation.
The objectives of this study were to evaluate the anatomy component in skin-separated
and normal date fruit and to assess the microclimatic conditions (temperature and relative
humidity) during the fruit developmental stages and its association with skin separation.
 2. Materials and Methods
2.1. Site Selection
This study was conducted at The Medjool Village farm, Jordan Valley, Jordan (long.
31° 58’41.14″ N, lat. 35° 33’2534″ W, elevation 325 m below sea level) from March 2021 to
October 2022 (Figure 1). The farm was awarded GLOBAL G.A.P. certification for applying
Agriculture 2023, 13, 38 3 of 11
international standards for crop production. The farm’s total area is about 128 ha, and the
total number of date palm trees (Phoenix dactylifera L.) var. “Mejhoul” (previously known
as “Medjool”) is about 17,700. Although several date palm cultivars (>10) are grown in
2. Materials
Jordan and Methods
including “Barhi” and “Khalas”, “Mejhoul” account for 62% of the total grown
2.1. Site Selection
area [11]. The study was carried out for two growing seasons in two selected sites at the
farm This
(Figurestudy wassoil
1). The conducted
chemicalatanalysis
The Medjool
for bothVillage
sites isfarm, JordaninValley,
presented Table 1.Jordan
Trees(long.
from
31◦ 58sites
both 0 41.1400 N, lat. 35◦ 330 253400 W, elevation 325 m below sea level) from March 2021 to
were about 8 years old and spaced 9 m within and between rows. Trees from
October
both sites2022
were(Figure 1). The
subjected farm was
to similar awarded GLOBAL
agricultural practices G.A.P.
including certification
irrigation,forfertilizers,
applying
and bunch thinning. Due to COVID‐19, workers removed only the lower one thirdand
international standards for crop production. The farm’s total area is about 128 ha, the
of the
total number
bunch of date
by cutting palm
back the trees (Phoenix
tips of dactylifera
the stands L.) var. “Mejhoul”
at pollination period. During(previously known
the growing
as “Medjool”)
seasons is about
(2020, 2021), the17,700. Although several
mean temperatures ranged date palm
from 33–34cultivars
°C and(>10) are humidity
relative grown in
Jordan including
between 35 and 46%. “Barhi” andirrigation
The total “Khalas”,per “Mejhoul”
tree per yearaccount
was 45 form62%
3 usingof athe total
drip grown
irrigation
area [11].
system. The study was carried out for two growing seasons in two selected sites at the
farm (Figure 1). The soil chemical analysis for both sites is presented in Table 1. Trees from
both sites
Table were
1. Soil about
analysis of8experimental
years old and spaced
sites at the 9Medjool
m within and between
Village, rows.Jordan.
Jordan Valley, Trees from both
sites were subjected to similar agricultural practices including irrigation, fertilizers, and
bunch thinning. Due to COVID-19, Electrical Conductiv‐
workers removed onlySoil CO2 Respira‐
the lower one thirdTotal
of theNbunch
Soil Texture PH (%)
ity (ds m −1) tion (mg
by cutting back the tips of the stands at pollination period. During the growing seasons kg −1)

Site2021),
(2020, 1 Clay‐loam 8.06
the mean temperatures ranged3.51 from 33–34 ◦ C and relative39.8 humidity1.07 between
35 Site
and 246%. Clay‐loam 8.01 per tree per2.89
The total irrigation year was 45 m using 3 33.6a drip irrigation 1.09
system.

Figure 1.
Figure Study sites
1. Study sitesatatThe
TheMedjool
MedjoolVillage,
Village,Jordan
JordanValley,
Valley,Jordan.
Jordan.Asterisk
Asterisk(*)
(*)represents
representsthe
thetree
tree
location
location (site 1 and 2) from which the fruit were sampled for anatomical observations.

Table 1. Soil analysis of experimental sites at the Medjool Village, Jordan Valley, Jordan.

Soil Texture PH Electrical Conductivity (ds m−1 ) Soil CO2 Respiration (mg kg−1 ) Total N (%)
Site 1 Clay-loam 8.06 3.51 39.8 1.07
Site 2 Clay-loam 8.01 2.89 33.6 1.09

2.2. Fruit Anatomy and Skin Separation

For the anatomical observations during the 2020 and 2021 growing seasons, 40 fruit of
each maturity stage [unripe (kimri and khlal) and ripe (Rutab and Tamr)] were collected
Agriculture 2023, 13, 38 4 of 11

(5 date palm trees per site). A 0.5 cm long section of fruit skin (with pulp) was taken. For each
mature and ripe stages, the fruit sections were immediately soaked in Formalin–Acetic Acid
fixative solution (FAA) following the procedure of Huang et al. [15]. 5 mm-thick transverse
sections were mounting on slides, dried, and stained in hematoxylin and eosin stain. Fruit
anatomy sections (cross and longitudinal) were analyzed using a light microscope (B-Scope
Trino E-plan, Euromax Microscopes Holland, Arnhem, The Netherlands) supplied with
a 12 pixel camera (CMEX 12, Euromax Microscopes Holland, Arnhem, The Netherlands).
Fruit anatomy variables were measured using image analysis software (Image Focus
Plus, Euromax Microscopes Holland, Arnhem, The Netherlands). Skin separation was
determined for both growing seasons and across sites (1 and 2) using fruit internal quality
system (TrueSort™, Ellips, Eindhoven, The Netherlands). The sorting system can accurately
detect internal defects in dates and is able to see completely through the exocarp.

2.3. Weather Data and Soil Moisture Content

Air temperature and relative humidity were measured continuously in three randomly
selected locations at both sites using external temperature/relative humidity data logger
(MX2302A, HOBO, Bourne, MA, USA). Sensors were installed on the top of the trees.
In addition, soil moisture content was measured daily at 11:00 a.m. using time-domain
reflectometry (TDR 100, Spectrum Technologies, Inc. Aurora, IL, USA).

2.4. Data Analysis

Skin separation data were analyzed using SAS statistical software (version 9.2; SAS
Institute, Cary, NC, USA). The analysis of variance (ANOVA) and the least significant
difference test (p < 0.05) in SAS were used to identify differences between treatments.

3. Results
The anatomy analysis of date palm fruits was carried out at the Kimri-Khalal, Khalal,
Rutab, and Tamr stages in 2020 (Figures 2 and 3). The sclereid cell numbers and clusters at
the early fruit developmental stage (Kimri-Khalal) were lower than the next succeeding
stages. However, the sclereid cluster area and perimeter were similar across the four stages,
except at Tamr in fruit with more than 50% skin separation (Figure 2). During the Kimri-
Rutab fruit developmental stages, the skin (cuticle + epidermis + hypodermis) thickness
ranged from 350–400 µm.
Table 2 shows the anatomical differences in date palm fruit ‘Mejhoul’ at the Tamr
stage in normal fruit and those in which skin separation exceeded 50%. In both years
(2020 and 2021), the sclereid cell number in normal fruit was significantly lower than in
skin-separated ones. In addition, the sclereid cluster number, area, and perimeter in normal
fruit was extremely lower than those that had the skin separation physiological disorder.

Table 2. Anatomical changes in date palm fruit ‘Mejhoul’ at the Tamr stage, September 2020 and 2021.

Distance
Sclereid Exocarp
Sclereid Sclereid between Cuticle Epidermis
Sclereid Cells Cells Cluster (Skin)
Fruit Quality (Skin) Clusters Cluster Sclereid Cells Thickness Thickness
(no mm−2 ) Perimeter Thickness
(no mm−2 ) Area (µm) Clusters and (µm) (µm)
(µm) (µm)
Cuticle (µm)
2020
Normal 78.4 b 6.66 b 34.7 b 680 b 67.3 a 3.97 a 13.2 a 232 a
Skin separation > 30% 105 a 8.20 a 47.4 a 848 a 44.4 b 3.60 a 11.2 b 213 a
p-value 0.007 0.02 0.013 0.02 <0.0001 0.19 0.01 0.57
2021
Normal 81.2 b 6.76 b 27.3 b 520 b 99.8 a 3.87 a 10.1 b 268 b
Skin separation > 30% 230 a 13.0 a 83.5 a 851 a 70.2 a 4.83 a 14.4 a 536 a
p-value <0.0001 0.003 0.001 0.008 0.13 0.14 0.001 0.004
Different letters in the same column indicate significant difference at p < 0.05.

Weather data (temperature and relative humidity), volumetric soil water content, and
irrigation during the fruit ripening stages (June–October, 2020 and 2021) are shown in
Figure 4. For site 1, mean air temperatures (2020, 2021) were between 33.7 and 33 ◦ C, and
Agriculture 2023, 13, 38 5 of 11

mean relative humidity was between 45.8 and 46%. Site 2 mean temperatures (2020, 2021)
were between 33.9 and 36 ◦ C, whereas mean relative humidity was between 36.3 and 41%.
The difference in temperatures between day and night at site 1 during the 2020 growing
season ranged from 12–21◦ C (mean 14.7 ◦ C) and from 11–21◦ C (mean 14.8 ◦ C) at site 2.
Differences in relative humidity (day/night) for 2020 ranged from 7 to 51% (mean 30%)
at site 1 and from 5 to 35% (mean 18%) at site 2. In 2021, temperature differences at site 1
ranged between 6.5–18.5◦ C (mean 12 ◦ C) at site 1 and 8.5–22 ◦ C (mean 16.8 ◦ C) at site 2. In
Agriculture 2023, 13, x FOR PEER REVIEW
addition, the difference in relative humidity between day and night in 2021 at site 51 of was12
between 13–46% (mean 29.6%) and 13.2–46.5% (mean 34%) at site 2.

Figure 2. Date palm fruit anatomy at (A) Kimri‐Khalal, (B) Khalal, (C) Rutab, and (D) Tamr with no
Figure 2. Date palm fruit anatomy at (A) Kimri-Khalal, (B) Khalal, (C) Rutab, and (D) Tamr with no
skin separation and (E) with skin separation >30%.
skin separation and (E) with skin separation >30%.
riculture 2023, 13, x FOR PEER REVIEW
Agriculture 2023, 13, 38 6 of 11

Figure 3. Sclereid total cell number, cluster number, area, and perimeter and exocarp (skin) thickness
of date palm fruit ‘Mejhoul’ at four developmental stages.
Figure 3. Sclereid total cell number, cluster number, area, and perimeter and exocarp
ness of date palm fruit ‘Mejhoul’ at four developmental stages.

Table 2 shows the anatomical differences in date palm fruit ‘Mejhoul’

stage in normal fruit and those in which skin separation exceeded 50%. In both
and 2021), the sclereid cell number in normal fruit was significantly lower t
separated ones. In addition, the sclereid cluster number, area, and perimete
fruit was extremely lower than those that had the skin separation physiologic

Table 2. Anatomical changes in date palm fruit ‘Mejhoul’ at the Tamr stage, Septem
2021.
lture 2023, Agriculture
13, x FOR PEER
2023, REVIEW
13, 38 8 of 12 7 of 11

Figure 4. Weather macroclimate,

Figure 4. Weather irrigation per tree,
macroclimate, and volumetric
irrigation soilvolumetric
per tree, and water content
soilduring the
water content during the
study period instudy
both sites
periodat the Medjool
in both sites Village, Jordan Village,
at the Medjool Valley, Jordan.
Jordan Valley, Jordan.

4. Discussion4. Discussion
4.1. Date Palm4.1. Date Palm
Industry Industry
and Skin and Skin Separation
Separation
Date fruits areDate fruits areavailable
commonly commonly available
in the globalinmarket,
the global market, predominantly
predominantly at the ma‐ at the mature
Tamr stage [4]. Although the date fruit is used in several food products worldwide, large
ture Tamr stage [4]. Although the date fruit is used in several food products worldwide,
amounts of dates ends up as waste [16]; this is approximately 20–30% of the total production
large amounts of dates ends up as waste [16]; this is approximately 20–30% of the total
in Jordan [12]. Low-quality date fruit (loss) has more fibers and a harder texture than the
production in Jordan [12]. Low‐quality date fruit (loss) has more fibers and a harder tex‐
high-grade ones [16]. In horticultural crops, the cuticle, epidermal, and hypodermal cells
ture than the high‐grade ones [16]. In horticultural crops, the cuticle, epidermal, and hy‐
represent the structural backbone of the skin during fruit development and ripening [17].
podermal cells represent the structural backbone of the skin during fruit development and
Interestingly, the visual appearance of a fruit is associated with mechanical characteristics
ripening [17]. Interestingly, the visual appearance of a fruit is associated with mechanical
of its skin as well as the skin integrity [17]. Overall, studies dealing with skin quality are
characteristics of its skin as well as the skin integrity [17]. Overall, studies dealing with
essential for date palm industries due to fact that the expected value of loss caused by the
skin quality are essential for date palm industries due to fact that the expected value of
skin physiological disorders (specifically, skin separation) is extremely high and could cost
Agriculture 2023, 13, 38 8 of 11

millions of dollars each year. In this study, the skin separation (>30% per fruit) ranged from
14.5% (2020) to 34% (2021) across the studied sites. The skin separation of ripe fruit is a
common physiological disorder which significantly reduces the quality of the fruit and,
consequently, its commercial value [18,19]. Generally, skin separation does not reduce the
nutritional value or taste of the fruit, but these symptoms do affect the visual appearance
of the fruit and consequently compromise its price value at the point of sale [20]. Therefore,
these disorders, including microcracking and separation, are of considerable commercial
importance [20]. Lustig et al. [19] concluded that when skin separation in the fruit exceeds
10%, it becomes disqualified for export. Considering the high percentage of skin separation
(this study: 14.5–32%, Jordan national statistics: 20–40%) of fruit yield, and due to its
negative effect on date marketing value (Jordan: ~$30 million loss), understanding the
main causes of this physiological disorder is of great interest to date palm growers.

4.2. Fruit Anatomy

In this study, the anatomical analysis of the date palm fruit revealed that the exocarp
or skin (cuticle, epidermis, hypodermis) and mesocarp thickness were quite similar across
the study period (2020, 2021), except for sclereid cells (Figures 2 and 3). During the ripening
stages (Kimri-Khalal, Khalal, Rutab and Tamr), the sclereid cell numbers and clusters
gradually increased and became prominent, especially in skin-separated fruit at the Tamr
stage. In fact, the anatomical analysis of the date revealed that the percentage increase
in sclereid cell number in skin-separated fruit at the Tamr stage ranged from 34–183%,
sclereid clusters number 23–92%, cluster area 37–206%, and cluster perimeter 25–64% when
compared to normal fruit (no skin separation), as shown in Table 2. In addition, the scle-
reid cells in skin-separated fruit were clustered together, forming a chain of aggregates
underneath the fruit skin, leading to the partial separation of exocarp from the flesh of
the fruit (mesocarp), as shown in Figure 2E. In pears (Pyrus communis, P. bretschneideri and
P. pyrifolia), stone (or sclereid) cells appeared in cluster structures 60 days after pollina-
tion [21]. In addition, these stone cells were clustered and moved closer to the epidermis
in P. pyrifolia pear than in P. communis and P. bretschneideri [21]. In our study, the distance
between cuticle and sclereid clusters in skin-separated fruit (compared to normal skin date)
was 42–50% shorter (Table 2). Overall, in this study, the skin separation of date fruits at
the Tamr stage occurred when the stone cells clustered, aggregated, and moved closer to
the exocarp.
Stone (sclereid) cells form sclerenchyma tissue that is developed by the deposition of
lignin onto the primary cell wall of parenchyma cells, leading to an increase in the cell wall
thickness [22,23]. High numbers of stone cells stimulate the development of rough skin fruit
and, thus, reduce fruit quality. Normally, stone cells’ numbers at early fruit developmental
stages are low but gradually increase at later stages, especially in rough-skin fruit [23]. In
loquat fruit, the content of stone cells can reach 1.6% (w/w) and size 1000 × 500 µm [22].
Stone cells content in sand pears (Pyrus pyrifolia) was between 2.8 and 29.0%, lignin was
between 8.8–55.3% and cellulose 11.5–30.6% [24]. Due to the high density of lignified stone
cells that were significantly correlated with the firmness and the lignin content of the bulk
flesh, Huang et al. [15] concluded that the increase in lignified cell number is a key factor that
reduces the quality of the postharvest loquat (Eriobotrya japonica) fruit. Therefore, in order
to identify the microclimatic stress factors that stimulate the development of rough-skinned
fruit, the analysis and assessment of physiological and molecular mechanisms of stone
cell formation is essential [23]. The main components of stone cells in horticultural crops
are lignin cellulose and hemicellulose, [23,25]. During cell development stages, lignin and
cellulose gradually fill the lignified cells, leading to significant morphological changes in
fruit [15]. In the date palm fruit, George et al. [25] postulated that parenchyma and sclereid
cells contain high levels of lignin and phytolith (inorganic particles of silicon dioxide).
The development of sclereids (stone cells) is closely related to the synthesis, transfer, and
deposition of lignin [26]. These sclereids are sclerenchyma cells that are developed by the
deposition of lignin on the primary walls of parenchyma cells [26]. During the deposition
Agriculture 2023, 13, 38 9 of 11

process, lignin molecules and microfibrils are alternatively arranged until they fill up the
whole cell cavity, ending in the development of stone cells [26]. In the Japanese pear fruit
(Pyrus serotina), the young sclereids formed secondary cell walls by the accumulation of
lignin [27]. This lignification process induced vesicular structures inside the cytoplasm,
hence the destruction of the cytoplasm [27].
Understanding the main structural organization of the networks formed by lignin
and silica (phytolith) in the fruit of the date palm is essential to assess their functional
roles in the mechanical strength and hardness as well as adaptation to biotic and abiotic
stresses [25]. The silicified plant tissue is named phytoliths [28], hence both lignin and
silica (phytolith) may contribute to the rigidity of date fruit [25]. Silica is an amorphous
(non-crystalline) material that does not establish specific bonds (antigen contact) with
biostructures, therefore, the rate of silicic acid polymerization and structure may be con-
trolled via silicic acid oligomers [28]. George et al. [25] found a significant heterogeneity
in date fruit silica phytoliths and the lignified structures. In their study, lignin presented
independently from both silica in the secondary cell walls of sclereid cells and the spheroid
phytoliths located around the sclereid cells. Spheroid phytoliths were abundant mainly
around the sclereid cells within the skin of the date fruit [25]. Bauer et al. [28] stated that the
bio-silicification in the plant involves the uptake of silicic acid from the soil solution, after
which it is condensed into solid silica (polymerized silica is one of the solidest materials
in the plant tissue), mainly in the epidermis. Bio-silicification positively correlates with
transpiration rates of the tissue (polymerization of the silica as water evaporates) [28]. Bio-
silicification in wheat was found in the epidermis cell wall and in sclerenchyma cells near
the vascular bundles, but not in the stomata, proposing that an active transport oriented
the soluble silica away from the water evaporation stream [29]. Silicon induces changes in
the mechanical properties of cell walls in plants. Silicon promotes the extensibility of cell
walls (cell elongation but not cell division) in the growing zone in the apical and subapical
zones by decreasing the mechanical hardening of these regions (elastic moduli and viscosity
coefficients) [30,31].

4.3. Weather Data

Microclimate conditions (temperature and relative humidity) during the pre-harvest
period affect fruit quality at the time of harvest and during post-harvest stages, specifi-
cally at long-term storage [9]. These conditions may result in changes in the composition
and cellular structures of the skin and sub-surface cellular layer, leading to significant
impact on fruit postharvest quality and storability [9]. Inconsistent or low relative humidity
at the ripening stage could induce some physiological disorders, including skin separa-
tion [2]. Most of these physiological disorders are often increased during fruit ripening and
postharvest operations, especially under high temperatures or relative humidity, leading to
potential reduction in fruit quality and severe economic losses [32]. For example, high rela-
tive humidity reduces the mechanical resistance of the cuticle and induces the development
of skin microcracks [32]. Transpiration is a physical process that occurs in ripening fruit
via the stomata and cuticle. Fruit transpiration rate correlated positively with temperature
and negatively with relative humidity [33]. Transpiration affects the water relations and
balance of the fruit skin. For example, in banana, the cumulative transpiration increased
linearly with the development of fruit stages [33]. However, unbalanced water relation
across the fruit layers may lead to fruit skin splitting and skin browning. In this study,
mean air temperatures for both growing seasons (2020, 2021) were between 33 and 34 ◦ C,
and mean relative humidity were between 45 and 46%. In addition, management practices
(irrigation, fertilization, pruning, etc.) were similar across the growing seasons (2020, 2021)
and in both sites (1 and 2). However, skin separation was found in 14 to 15% of the total
fruit in 2020, and 30 to 34% in 2021 growing season. Although the mean temperature and
relative humidity as well as management practices were similar across the seasons, the skin
separation amount of total yield was potentially higher in 2021 than 2020. These results
indicate that the weather conditions, specifically temperatures and relative humidity, are
Agriculture 2023, 13, 38 10 of 11

not the only factors that stimulate the skin separation physiological disorder in date palm
fruits. Overall, further studies are required to identify the main factors that stimulate the
development of sclereid cells and the potential of agricultural practices.

5. Conclusions
In conclusion, in both growing seasons and across the study sites, the number, clusters,
and aggregate of stone (sclereid) cells increased during the fruit ripening stages (khimri to
Tamr), specifically, in skin-separated fruit at the Tamr stage. In addition, these sclereid cells
were clustered and moved closer to the exocarp (skin). However, although the microclimate
conditions (mean temperatures and relative humidity) were similar across the growing
seasons (2020, 2021), the skin separation percentage in fruit yield in 2020 ranged from 14 to
15% and in 2021 from 30 to 34%; this demonstrates that the weather conditions are not the
only components that induce the development of skin separation in the date palm fruit. A
future study is intended to explore the possible interactive effects of management practices
and the micro-environment with the abrupt increase of sclereid clusters and aggregates in
skin-separated fruit as well as the chemical and mechanical assessment of fruit skin.

Author Contributions: Conceptualization, N.A. and Y.O.; methodology, N.A., N.E.-A. and Y.O.;
software, M.A.-A. and J.A.; validation, N.A., Y.O. and N.E.-A.; formal analysis, N.A. and Y.O.; investi-
gation, N.A. and Y.O.; data curation, N.A., M.A.-A. and Y.O.; writing—original draft preparation, N.A.
and Y.O.; writing—review and editing, N.E.-A., J.S. and J.A.; supervision, Y.O.; project administration,
N.A.; funding acquisition, N.A. and Y.O. All authors have read and agreed to the published version
of the manuscript.
Funding: This research was funded by the Deanship of Scientific Research at the University of Jordan,
grant number 2366.
Institutional Review Board Statement: Not applicable.
Data Availability Statement: Not applicable.
Acknowledgments: We thank the Medjool Village farm, Jordan Valley, for supplying date palm
fruits and conducting our study at their farm. Appreciation is also extended to Engs. Yazan Nabulsi,
Mohammad Al-Edwan, Ehab Al-Balawneh, Ibrahim Al-Bakeet and Tala A’saf for their assistance in
the field and laboratory.
Conflicts of Interest: The authors declare no conflict of interest.

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