Postharvest Biology and Technology 142 (2018) 39–45

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Postharvest Biology and Technology

journal homepage: www.elsevier.com/locate/postharvbio

Characterization of stomata on floral organs and scapes of cut ‘Real’ gerberas T

and their involvement in postharvest water loss
⁎
Xinmin Huanga,b, Shuqin Lina, Shenggen Hea, , Xiaohui Lina, Jiping Liua, Riyuan Chenb,
⁎
Hongmei Lia,
a
College of Agriculture and Biology, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, PR China
b
Guangdong Provincial Engineering Technology Research Center for Protected Horticulture, College of Horticulture, South China Agricultural University, Guangzhou
510642, PR China

A R T I C LE I N FO A B S T R A C T

Keywords: Stomata present on leaves are the main channels for water vapor exchange between the plant and atmosphere,
Cut gerberas and contribute to water loss of cut flowers. Although cut gerberas (Gerbera jamesonii Bolus) are commonly
Sepal composed of a terminal composite floral head and a scape, and have no leaves, they are also prone to severe
Stomatal distribution water deficits, presumably resulting from water loss exceeding water uptake. We have investigated the dis-
Stomatal opening
tribution and characteristics of stomata of cut ‘Real’ gerberas, and their response to light and abscisic acid (ABA)
Water loss
treatments, to increase understanding of their functionality in postharvest water loss. Stomata were widely
distributed on various parts of floral organs and the scape in cut gerberas, including the lower epidermis of the
ray floret petals, trans florets, disc florets, lower epidermis of sepals, and the uppermost and middle parts of
scape surfaces. The highest stomatal density was observed on the lower epidermis of sepals. Cut gerberas had the
largest stomata on the middle scape surface and the smallest on the surface of the uppermost scape. Nonetheless,
the lower epidermis of sepals had the highest relative stomatal area, which was higher than that of the other
parts. Additionally, the lower sepal epidermis had the highest maximum stomatal conductance of water vapor
(gwmax), and which was higher than that of the lower and upper leaf epidermis. The opening of stomata on
various parts of cut gerberas differed in their response to light, and the highest percentage of open stomata and
largest aperture were found in the stomata on the lower epidermis of sepals. Changes in water loss in cut
gerberas exhibited a distinct circadian rhythm over a four-day vase period with an alternating 12-h light/dark
cycle. In the lower epidermis of sepals, ABA painting treatment significantly inhibited stomatal opening and
reduced water loss in cut gerberas over a 6 h light period. These findings imply the critical involvement of
stomata on the lower epidermis of sepals in water loss of cut gerberas, and improve our understanding of the
mechanism underlying their water deficit symptoms.

1. Introduction et al., 2006).

Gerbera (Gerbera jamesonii Bolus), commonly grown as a cut-flower
crop, belongs to the family Asteraceae. This species is well known for its
abundant and diverse flower colors, sizes, and shapes, and it has been
consistently ranked among the top six most popular cut flowers
worldwide (Bhatia et al., 2009; Frómeta et al., 2017). A cut gerbera is
normally composed of a terminal composite floral head (inflorescence)
and a stem (scape) that does not have leaves (Perik et al., 2012). Ad-
ditionally, unlike species with a single type of flower, the gerbera in-
florescence has three types of florets: ray, trans, and disc florets, which
are tightly condensed at the same receptacle (Kuang et al., 2013; Teeri
Although cut gerberas have no leaves, the floret petals often wilt
and the scape can bend or break during storage and display periods (Liu
et al., 2009; Perik et al., 2012, 2014). For many cut flowers, including
cut gerberas, the development of such water deficit symptoms is mainly
attributed to the water loss gradually surpassing water uptake (He
et al., 2006; Mattos et al., 2017; Perik et al., 2012, 2014; van Doorn,
2012). Therefore, both increased water uptake and decreased water loss
are important approaches to alleviate the water deficit in cut flowers
(de Witte et al., 2014; Fanourakis et al., 2016; Liu et al., 2009; Mattos
et al., 2017; van Doorn, 1997, 2012).
Water loss in cut flowers occurs mainly via stomatal transpiration

⁎
Corresponding authors.
E-mail addresses: hxmscau@163.com (X. Huang), shuqinzhku@163.com (S. Lin), shenggenzhku@163.com (S. He), xiaohuizhku@126.com (X. Lin), liujipingpipi@163.com (J. Liu),
rychen@scau.edu.cn (R. Chen), lihongmei0000@163.com (H. Li).

https://doi.org/10.1016/j.postharvbio.2018.04.001
Received 20 November 2017; Received in revised form 2 April 2018; Accepted 2 April 2018
Available online 11 April 2018
0925-5214/ © 2018 Elsevier B.V. All rights reserved.
X. Huang et al.
(Aliniaeifard and van Meeteren, 2016; Carvalho et al., 2015; In et al.,
2016). The stomata are specialized pores in the epidermis that are
bound by two guard cells. Water loss occurs when stomata open, which
is driven by volume variations in the guard cells (Mori and Murata,
2011). Stomatal opening can cause excessive water loss and serious
water deficits, thereby, leading to early wilting and rapid senescence in
cut flowers (Farrell et al., 2012; Halevy and Mayad, 1981; van Doorn,
2012). Most water loss in cut roses occurred through open stomata on
leaves and resulted in enhanced wilting of petals and leaves, as well as
in the bent-neck phenomenon (Fanourakis et al., 2012; Slootweg and
van Meeteren, 1991). However, suppression of stomatal opening by
keeping the flowers in darkness (Fanourakis et al., 2013) or treatments
with various compounds, such as abscisic acid (ABA) (Kitamura and
Ueno, 2015; Kohl and Rundle, 1972), sodium nitroprusside as a nitric
oxide donor (Fanourakis et al., 2016), and acetylsalicylic acid
(Fanourakis et al., 2016) can reduce water loss and alleviate water
deficit symptoms, thereby, extending the display life of cut flowers.
Most studies on how stomata are involved in water loss in cut
flowers have mainly focused on the leaves, e.g., roses (Fanourakis et al.,
2013, 2016; Lü et al., 2010), carnations (Carpenter and Rasmussen,
1974), chrysanthemums (Aliniaeifard and van Meeteren, 2016), and
snapdragons (Schroeder and Stimart, 2005). Nonetheless, stomata are
also distributed in parts other than leaves in some cut flowers, such as
carnation stems (Carpenter and Rasmussen, 1974), tulip petals (Azad
et al., 2007), anthurium spathes (Elibox and Umaharan, 2008, 2010),
and hydrangea sepals (Kitamura and Ueno, 2015). In cut gerberas,
which have no leaves, a high transpiration rate was detected during
their postharvest period (Perik et al., 2012; van Meeteren, 1978), which
led to the hypothesis that stomata are distributed on their non-leaf
organs, and can contribute to water loss.
Little is known about stomatal distribution on non-leaf parts of cut
gerberas and their characteristics, and their functionality in water loss.
Hence, the aim of this study was to investigate the distribution of sto-
mata on various parts of the floral organs and scapes of cut ‘Real’
gerberas, and to characterize the stomatal density, size, and response to
light and ABA treatments, in addition to deciphering their association
with postharvest water loss.
2. Materials and methods
2.1. Plant material
Flowering stems and fully expanded leaves were freshly harvested
from gerbera plants (Gerbera jamesonii Bolus ‘Real’) grown in the ex-
perimental field at the Zhongkai University of Agriculture and
Engineering to observe stomatal distribution, density, and size para-
meters.
Cut gerbera flowering stems were purchased from a wholesale cut-
flower market in Guangzhou City, P.R. China. The flowering stems were
stood upright in plastic buckets partially filled with tap water and then
transported within 1 h to the laboratory. To minimize water loss and to
prevent mechanical damage, the flowering stems were covered with a
sheet of plastic film during transportation. Upon arrival, the flowering
stems were selected for their uniformity in size, color, and freedom
from defects by careful visual inspection, and were re-cut under deio-
nized water (DIW) to an approximately 25 cm length.
2.2. Experimental design and treatments
All the experiments were performed in a phytotron, operating at
20 ± 2 °C, under 60 ± 10% relative humidity (RH) and
100 μmol m−2 s−1 light intensity (LED light, Guangzhou Blueseatec Co.
Ltd.). The cut gerberas were held individually in 150 mL glass vases
containing 100 mL DIW.
40
Postharvest Biology and Technology 142 (2018) 39–45
2.2.1. Observation of stomatal distribution, density, and size parameters
To investigate the stomatal distribution, density, and size in the disc
florets, trans florets, ray florets, sepals, scape, and detached leaves were
prepared for scanning electron microscopy (SEM) (see Section 2.3).
2.2.2. Effects of light on stomatal opening
The cut gerberas were held in DIW for 3 h in the dark. Subsequently,
the cut gerberas were incubated under light (100 μmol m−2 s−1) for 7 h,
and various parts of their floral organs and scapes were sampled from
three different flowering stems to measure the percentage of open
stomata and stomatal apertures by SEM.
2.2.3. Water loss under alternating light and dark conditions
The cut gerberas were held in DIW and incubated under a 12 h light
(100 μmol m−2 s−1) and 12 h dark cycle for 96 h, and water loss data
were collected at 1 h intervals.
2.2.4. Effects of ABA on sepal stomatal opening and water loss in cut
gerberas
The cut gerberas were held in DIW for 3 h in the dark. Subsequently,
the lower epidermis of the sepals were painted with 100 μmol L−1 ABA
(≥98.5%, Sigma-Aldrich, Steinheim, Germany, which was prepared by
dissolving 0.0264 g ABA in 0.5 mL 95% alcohol, and then diluted with
DIW to a final volume of 1000 mL) or DIW (with equivalent alcohol) as
control. The control and ABA-treated cut gerberas were individually
held in vases containing 100 mL DIW in the light (100 μmol m−2 s−1)
for 6 h. Thereafter, the sepals from the control and ABA-treated cut
gerberas were sampled for SEM to investigate the percentage of open
stomata and stomatal apertures. Concurrent with the observations and
measurements for stomatal opening, water loss data for cut gerberas
were collected at intervals of 10 min (see Section 2.4 and 2.5). Three
biological replicates were used for each treatment (i.e., three samples
were observed for each treatment).
2.3. SEM imaging of stomata
All the samples were cut into small sections (approximately
2 mm × 5 mm) and immediately fixed in 4% (v/v) glutaraldehyde in
0.1 mol L−1 phosphate buffer (pH 6.8) at 4 °C for 36 h, according the
method of Zobayed et al. (2001) with slight modification. The samples
were dehydrated in a graded series of 30, 50, 70, 85, 95, and 100%
ethanol solutions. The samples were then subjected to CO2 critical point
drying and gold sputter coating. The samples were observed at 10 kV
accelerating voltage using a JSM-6360LV SEM (JEOL Ltd., Tokyo,
Japan) and were photographed.
2.4. Characterization of stomata
2.4.1. Stomatal density
For each sample from the different parts of gerbera plants, the
stomatal density (SD, number of stomata per mm2) of the lower and
upper epidermis of floral organs and leaves, along with the surfaces of
the upper and middle parts of scapes, was determined as described by
Balasooriya et al. (2009).
2.4.2. Stomatal size parameters
Stomatal area (SA, μm2) and relative stomatal area (RSA, %) were
calculated according to the following equations (Orcen et al., 2013;
Pompelli et al., 2010):
Wg Lg
SA = π × ×
2 2
RSA = SA × SD × 100
where, guard cell length (Lg, μm) and width (Wg, μm) were defined as
the longest axis (length) and widest point perpendicular to that axis
X. Huang et al. Postharvest Biology and Technology 142 (2018) 39–45

(width) of the guard cell, respectively.

2.4.3. Maximum stomatal conductance to water vapor (gwmax)

The maximum stomatal conductance to water vapor (gwmax,
mol m−2 s−1) in fully opened stomata from different parts of cut ger-
bera flower stems was calculated according to the following equation
(Franks and Beerling, 2009):
d π
g wmax = × SD × αmax /(l + αmax / π )
v 2
where, d is the diffusion of water vapor in the air (24.9 × 10−6 m2 s−1)
at 25 °C, v is the molar volume of air (24.4 × 10−3 m3 mol−1) at 25 °C,
αmax is the maximum area of the open stomatal pore, and l is stomatal
pore depth. αmax was approximated as π (p/2)2, where p is stomatal
pore length, which was assumed to be Lg/2 (Franks and Beerling,
2009). The pore depth (l) for fully open stomata was equal to Wg/2,
assuming guard cells inflate to a circular cross-section (Franks and
Beerling, 2009; Milla et al., 2013).

2.4.4. Percentage of open stomata

The percentage of open stomata was determined according to the
following equation (Chen and Gallie, 2004):
no
Percentage of open stomata = × 100
nt
where, no is the number of open stomata, nt is the total number of
stomata.

2.5. Water loss from cut gerbera flower stems

The water loss from cut gerberas was determined using a continuous
automatic apparatus as described by Lü et al. (2011).

2.6. Statistical analysis

Sigmaplot 11.0 software (Systat Software Inc., San Jose, CA, USA)
was used to analyze the data with one-way analysis of variance
(ANOVA). The Duncan’s multiple range test at the 0.05 level was used
to detect significant differences among the stomatal characteristics in
different parts of cut gerberas. The data are presented as mean ±
standard error (SE).

3. Results Fig. 1. Stomatal distribution on the different parts on the floral organs and
scapes of cut ‘Real’ gerberas.
Stomata present on the (A) lower epidermis of ray floret petals; (B) lower
3.1. Stomatal distribution
epidermis of trans floret petals; (C) lower epidermis of disc floret petals; (D)
lower epidermis of sepals; (E) surface of the uppermost part of the scape; and
Stomata were observed on various parts of floral organs and scapes (F) surface of middle part of the scape. Scale bar = 100 μm, scale bar in the
of cut ‘Real’ gerbera stems, including the lower epidermis of petals on inset figure = 25 μm.
ray floret (Fig. 1A), trans florets (Fig. 1B), disc florets (Fig. 1C), lower
epidermis of sepals (Fig. 1D), and the surfaces of the upper and middle
different parts of floral organs and scapes of cut gerberas (Table 1). The
parts of scapes (Fig. 1E and F). However, no stomata were found on the
Lg and Wg, and SA values of cut gerberas were the largest on the surface
upper epidermis of petals on trans florets, ray florets, and sepals.
of the middle part of the scape and smallest on the surface of the up-
permost part of the scape. Stomata on the floral organs, including the
3.2. Stomatal density, size, and gwmax
lower epidermis of sepals, lower epidermis of petals of ray florets, trans
florets, and disc florets, were of an intermediate size, not significantly
There were considerable differences in stomatal density on various
different from one another. However, the lower sepal epidermis had the
parts of floral organs and scapes of cut gerberas (Table 1). The number
highest RSA value, which was significantly higher than that of the other
of stomata on cut gerberas, arranged in decreasing order, was found on
parts of cut gerberas studied, including the leaves.
the lower epidermis of sepals (205 stomata mm−2), the surface of the
The gwmax values for the different floral organs and scapes of cut
uppermost part of the scape (160 stomata mm−2), lower epidermis of
gerberas were investigated (Table 1). For example, the lower epidermis
the disc floret petals (155 stomata mm−2), the lower epidermis of trans
of the sepals had the highest gwmax value (2.31 mol m−2 s−1), which
floret petals (60 stomata mm−2), the surface of middle part of the scape
was significantly higher than those measured in stomata on the lower
(70 stomata mm−2), and lower epidermis of ray floret petals (75 sto-
and upper epidermis of leaves (1.97 and 0.78 mol m−2 s−1, respec-
mata mm−2). In contrast, the stomatal density on the lower and upper
tively). The lower epidermis of trans floret petals had the lowest gwmax
epidermis of leaves was 260 and 95 stomata mm−2, respectively.
value (0.68 mol m−2 s−1), and gwmax measured in other parts of cut
There were differences in the sizes of stomata present on the

41
X. Huang et al. Postharvest Biology and Technology 142 (2018) 39–45

Table 1
The stomatal density, size parameters, and maximum stomatal conductance to water vapor (gwmax) in the different floral organs and scapes of cut ‘Real’ gerberas as
compared with those in leaves. Data presented for stomatal density are the mean ± SE based on five random observation areas, and data presented for stomatal size
parameters and gwmax are the mean ± SE of 20 stomata in five random observation areas. Small letters accompanying the values indicate significant differences at
P ≤ 0.05.
Parts Stomatal density (stomata mm−2) Stomatal size parameters gwmax (mol m−2 s−1)

Lg (μm) Wg (μm) SA (μm2) RSA (%)

LER 75 ± 7.9 de 41.06 ± 0.69 b 21.88 ± 0.62 c 709.48 ± 29.67 b 5.32 ± 0.22 de 0.94 ± 0.08 d
LET 60 ± 6.1 e 39.71 ± 0.78 b 24.63 ± 0.40 a 770.34 ± 24.57 b 4.62 ± 0.15 ef 0.68 ± 0.08 f
LED 155 ± 14.6 c 40.64 ± 0.72 b 22.08 ± 0.40 c 705.67 ± 20.49 b 10.94 ± 0.32 b 1.90 ± 0.20 b
LES 205 ± 14.6 b 39.44 ± 0.78 b 24.58 ± 0.62 a 764.77 ± 30.30 b 15.68 ± 0.62 a 2.31 ± 0.25 a
SUS 160 ± 10.0 c 33.98 ± 0.80 c 22.88 ± 0.51 bc 614.38 ± 25.71 c 9.83 ± 0.41 c 1.50 ± 0.19 c
SMS 70 ± 5.0 de 46.90 ± 0.77 a 24.28 ± 0.47 ab 895.77 ± 26.88 a 6.27 ± 0.19 de 1.01 ± 0.10 d
UEL 95 ± 5.0 d 28.45 ± 0.41 d 17.67 ± 0.51 e 384.13 ± 13.78 d 3.65 ± 0.13 f 0.78 ± 0.08 e
LEL 260 ± 12.5 a 27.79 ± 0.57 d 19.17 ± 0.56 d 421.23 ± 19.08 d 10.95 ± 0.50 b 1.97 ± 0.21 b

LER: Lower epidermis of ray floret petals; LET: Lower epidermis of trans floret petals; LED: Lower epidermis of disc floret petals; LES: Lower epidermis of sepals; SUS:
Surface of the uppermost part of the scape; SMS: Surface of the middle part of the scape; UEL: Upper epidermis of leaves; LEL: Lower epidermis of leaves. Lg: guard
cell length; Wg: guard cell width; SA: Stomatal area; RSA: Relative stomatal area; gwmax: Maximum stomatal conductance to water vapor.

gerberas ranged between 0.94 and 1.90 mol m−2 s−1.

3.3. Stomatal opening in response to light

The opening of stomata on the various parts of floral organs and

scapes of cut gerberas differed significantly in response to light
(Table 2). Under the given light conditions, the lower epidermis of
sepals had the highest percentage of open stomata, which correlated
with the largest stomatal aperture measurements. The lower epidermis
of disc floret petals had the lowest percentage of open stomata and the
smallest stomatal aperture measurements. The surfaces of the upper-
most and middle scape and the lower epidermis of petals of trans florets
and ray florets had an intermediate percentage of open stomata and
stomatal aperture measurements, and there were no significant differ-
ences among these parts. Overall, the stomata in the green parts of cut
gerberas, such as the sepals and scapes, were the most responsive to
light as compared with the other parts.
Fig. 2. Water loss rate in cut ‘Real’ gerberas observed over four days in an
alternating 12-h light/dark cycle. Water loss of cut gerbera stem was measured
3.4. Changes in water loss under alternating light and dark periods with a continuous automatic apparatus at intervals of 1 h. The open and solid
blocks indicate light and dark periods, respectively.
Changes in the water loss in cut gerberas exhibited a distinct cir-
cadian rhythm over a four-day 12 h light/dark period (Fig. 2). The 3.5. Stomatal opening of sepals and water loss in response to ABA treatment
water loss rate increased gradually in the light and declined in the dark,
with a peak and valley at the middle each light and dark period, re- The SEM imaging showed that the painting treatment with
spectively. 100 μmol L−1 ABA reduced stomatal opening in the lower epidermis of
sepals of cut gerberas under the light, as compared with that in the
Table 2 DIW-treated control (Fig. 3A and B). In addition, the percentage of open
Percentage of open stomata and stomatal apertures measured in the different stomata and the stomatal aperture in the ABA-treated sepals were lower
parts of floral organs and scapes of cut ‘Real’ gerberas that were incubated in than that in the control sepals (Fig. 3C). Moreover, the accumulated
light conditions (100 μmol m−2 s−1) for 7 h. The value of percentage of open
water loss of the ABA-treated sepals was consistently lower than that of
stomata represents the mean ± SE of three random observation areas, and
the control over the 6 h light incubation period (Fig. 3D).
value of stomatal aperture represents the mean ± SE of 20–50 stomata in three
random observation areas. Small letters accompanying the values indicate
significant differences at P ≤ 0.05.
4. Discussion
Parts Open stomata (%) Stomatal aperture (μm)
In addition to the leaves, stomata are also distributed on other plant
LER 41.1 ± 8.4 bc 0.72 ± 0.20 bc
LET 41.9 ± 1.0 bc 0.82 ± 0.20 bc tissues in addition to leaves in some cut flowers (Azad et al., 2007;
LED 25.7 ± 8.7 c 0.30 ± 0.08 c Carpenter and Rasmussen, 1974; Elibox and Umaharan, 2008, 2010;
LES 77.0 ± 6.1 a 2.00 ± 0.25 a Kitamura and Ueno, 2015). In the present study, we used SEM imaging
SUS 63.9 ± 4.5 ab 0.98 ± 0.13 b to show that stomata are widely distributed on different parts of floral
SMS 57.8 ± 8.9 ab 0.98 ± 0.26 b
organs and scapes of cut ‘Real’ gerberas (Fig. 1), including the lower
LER: Lower epidermis of ray floret petals; LET: Lower epidermis of trans floret epidermis of petals from ray florets, trans florets, disc florets, the lower
petals; LED: Lower epidermis of disc floret petals; LES: Lower epidermis of se- epidermis of sepals, and on the surfaces of the uppermost and middle
pals; SUS: Surface of the uppermost part of the scape; SMS: Surface of the scape. No stomata were observed on the upper epidermis of trans
middle part of the scape. florets, ray florets, or sepals. These results indicated that the various

42
X. Huang et al.
parts of the floral organs and scapes of cut gerberas are hypostomatic.
In contrast, the leaves of cut gerberas are amphistomatous, such that
stomata distributed on the lower and upper epidermis of leaves
(Table 1), which is consistent with previous observations (Romero-
Aranda et al., 1994).
The stomatal morphological characteristics, such as density and
size, are important for regulating gas exchange and transpiration in the
leaves of plants (Franks et al., 2009; Franks and Beerling, 2009;
Hetherington and Woodward, 2003). Similarly, for cut flowers, these
indices have important implications for their contribution to post-
harvest water vapor exchange (Carvalho et al., 2015; Fanourakis et al.,
2012; In and Lim, 2018). Schroeder and Stimart (2005) reported that
short-lived snapdragon genotypes had leaves with high stomatal den-
sity, while long-lived genotypes had leaves with low stomatal density.
Additionally, roses grown in high humidity environments had a
43
Postharvest Biology and Technology 142 (2018) 39–45
Fig. 3. Effects of painting treatment with 100 μmol L−1 abscisic
acid (ABA) of the lower epidermis of sepals on stomatal opening
and water loss in the cut ‘Real’ gerberas over a 6-h light
(100 μmol m−2 s−1) period. A and B: Images of stomata in the
control (DIW) and ABA-treated sepals after a 6 h light, respec-
tively. Scale bar = 25 μm. C: Percentage of open stomata (Left)
and stomatal apertures (Right) of control and ABA-treated sepals
after a 6 h light period. Value of percentage of open stomata re-
presents the mean ± SE of three random observation areas, and
value of stomatal aperture represents the mean ± SE of 30 sto-
mata in three random observation areas. D: Accumulated water
loss of three cut gerbera stems in the control and ABA-treated
sepals, measured with a continuous automatic apparatus at 10 min
intervals. The dynamic synchronous data are presented as a
means ± SE of three replicates.
significantly higher number of stomata and longer stomata as compared
to roses grown in low humidity, which resulted in enhanced water loss
and faster wilting (Fanourakis et al., 2013; Torre and Fjeld, 2001; Torre
et al., 2003). In the current study, significant differences were found in
the density and size of stomata on the various parts of floral organs and
scapes of cut gerberas (Table 1). Among all the parts tested, the lower
epidermis of sepals had the highest stomatal density, although it was
lower than that of the lower epidermis of leaves. Moreover, the lower
epidermis of sepals had the highest relative stomatal area, which was
higher than that of the other parts of cut gerberas, including the leaves.
The theoretical anatomical maximum stomatal conductance to water
vapor (gwmax) was derived from the stomatal size and density (Franks
et al., 2009; Franks and Beerling, 2009; Luomala et al., 2005). We found
that the lower epidermis of sepals possessed the highest gwmax value of
the parts tested, which was higher than that of the lower and upper
X. Huang et al.
epidermis of leaves. These findings indicate that stomata on the lower
epidermis of sepals may play a critical role in water loss in cut gerberas.
For many cut flowers, water loss is mainly attributed to stomata
opening on their leaves (Aliniaeifard and van Meeteren, 2016;
Fanourakis et al., 2013, 2016; Lü et al., 2010; Schroeder and Stimart,
2005). The studies on the contribution of stomata on the non-leaf or-
gans of some cut flowers to water loss have been also reported; how-
ever, the stomata on the non-leaf organs of cut flowers are not always
functional (Elibox and Umaharan, 2008, 2010; Hew et al., 1980;
Kitamura and Ueno, 2015; van Doorn, 1997). Carpenter and Rasmussen
(1974) observed a small number of stomata on cut rose ‘Forever Yours’
stem surfaces that appeared to be non-functional. Hew et al. (1980)
reported that stomata on tepals of orchids (Dendrobium biggibbum
Lindl.) failed to respond to light and ABA treatments, suggesting that
those stomata contributed little to transpiration control in these
flowers. Additionally, almost all of the stomata on spathes of cut an-
thuriums (Anthurium andraeanum Hort.) were closed, and there was no
correlation between their stomatal density and vase life (Elibox and
Umaharan, 2010). Similarly, most of the stomata on the decorative
sepals of cut hydrangea (Hydrangea spp. ‘Endless Summer’) flowers were
closed under light conditions, and only low stomatal conductance was
detected on the abaxial epidermis of their sepals, indicating that most
stomata on the decorative sepals were not involved in the postharvest
transpiration (Kitamura and Ueno, 2015). Nonetheless, the stomata on
the non-leaf organs of other cut flowers are associated with postharvest
transpiration. Cut carnations (‘White Sim’) had large numbers of sto-
mata on stem surfaces, which contributed to their water uptake and
water loss during the vase period (Carpenter and Rasmussen, 1974).
Azad et al. (2007) reported that stomata on tulip petals could affect
water transpiration during flower opening and closing.
In the case of cut gerberas, which typically presented without
leaves, had a high transpiration rate during postharvest life (Perik et al.,
2012; van Meeteren, 1978). However, there is little known about the
mechanism of water loss from cut gerberas. To test whether the stomata
distributed on the non-leaf organs of cut gerberas are involved in their
water loss, we further investigated the functional characteristics of
stomata on the various parts of floral organs and scapes. All stomata
observed responded to light, although their responses differed among
tissues in terms of the percentage of open stomata and stomatal aper-
ture (Table 2). Overall, stomata on the green parts of cut gerberas, such
as sepals and scape, were more responsive to light than the stomata
measured in other, non-green tissues. Nevertheless, stomata on the
lower epidermis of sepals had the highest percentage of open stomata
and largest stomatal aperture under light, whereas stomata on the scape
had a high percentage of open stoma but smaller stomatal aperture than
the stomata present on other tissues tested (Table 2). Further ob-
servation by SEM demonstrated that treating the lower epidermis of
sepals with 100 μmol L−1 ABA inhibited stomatal opening under light
(Fig. 3A and B). Additionally, the percentage of open stomata and the
stomatal aperture of ABA-treated sepals were lower than those of DIW-
treated control (Fig. 3C). The strong response of stomata present on
sepals to light and ABA suggested that they might be a key contributor
to water loss of cut gerberas.
In addition, the change of water loss in cut gerberas exhibited a
typical circadian rhythm, i.e., it increased in the light and decreased in
the dark (Fig. 2), consistent with what has been shown in cut flowers
with leaves (Doi et al., 1999; Lü et al., 2011). This phenomenon is due
to light-induced stomatal opening, which increases water loss in cut
flowers (Wang et al., 2017; van Doorn, 1997, 2012). Moreover, it was
found that the accumulated water loss in ABA-treated cut gerberas was
consistently lower than that in control plants over the 6 h light period
(Fig. 3D). These findings further imply that stomata on sepals are cri-
tically involved in water loss of cut gerberas.
44
Postharvest Biology and Technology 142 (2018) 39–45
5. Conclusion
Stomata are widely distributed on various parts of floral organs and
scape of cut gerberas. The highest gwmax value, percentage of open
stomata and apertures were observed in stomata on the lower epidermis
of sepals. Furthermore, we observed their response to light and ABA
treatments towards their functionality in water loss, which implies the
non-leaf stomata, particularly stomata on the lower epidermis of sepals,
are involved in water loss of cut gerberas. These findings would help to
improve our understanding of the mechanism underlying their water
deficit symptoms. Moreover, some physical or chemical approaches to
inhibit the opening of stomata on the non-leaf parts of cut gerberas,
particularly on sepals, are strongly suggested for further testing, with
the aim of establishing a practical and effective method for their post-
harvest treatment and preservation.
Funding
This work was supported by the Natural Science Foundation of
China (grant numbers 31672180, 31272193, and 31401897) and the
Natural Science Foundation of Guangdong Province (grant numbers
2016A030313374, 2014A03011027).
Conflicts of interest
There are no conflicts of interest to declare.
Acknowledgments
The authors thank Mrs. Xiaoying Hu from the South China Botanical
Gardens, for assistance with scanning electron microscopy.
References
Aliniaeifard, S., van Meeteren, U., 2016. Stomatal characteristics and desiccation re-
sponse of leaves of cut chrysanthemum (Chrysanthemum morifolium) flowers grown at
high air humidity. Sci. Hortic. 205, 84–89. http://dx.doi.org/10.1016/j.scienta.2016.
04.025.
Azad, A.K., Sawa, Y., Ishikawa, T., Shibata, H., 2007. Temperature-dependent stomatal
movement in tulip petals controls water transpiration during flower opening and
closing. Ann. Appl. Biol. 150, 81–87. http://dx.doi.org/10.1111/j.1744-7348.2006.
00111.x.
Balasooriya, B., Samson, R., Mbikwa, F., Boeckx, P., van Meirvenne, M., 2009.
Biomonitoring of urban habitat quality by anatomical and chemical leaf character-
istics. Environ. Exp. Bot. 65, 386–394. http://dx.doi.org/10.1016/j.envexpbot.2008.
11.009.
Bhatia, R., Singh, K.P., Jhang, T., Sharma, T.R., 2009. Assessment of clonal fidelity of
micropropagated gerbera plants by ISSR markers. Sci. Hortic. 119, 208–211. http://
dx.doi.org/10.1016/j.scienta.2008.07.024.
Carpenter, W.J., Rasmussen, H.P., 1974. The role of flower and leaves in cut flower water
uptake. Sci. Hortic. 2, 293–298. http://dx.doi.org/10.1016/0304-4238(74)90038-7.
Carvalho, D.R., Koning-Boucoiran, C.F., Fanourakis, D., Vasconcelos, M.W., Carvalho,
S.M., Heuvelink, E., Krens, F.A., Maliepaard, C., 2015. QTL analysis for stomatal
functioning in tetraploid Rosa × hybrida grown at high relative air humidity and its
implications on postharvest longevity. Mol. Breed. 35, 172. http://dx.doi.org/10.
1007/s11032-015-0365-7.
Chen, Z., Gallie, D.R., 2004. The ascorbic acid redox state controls guard cell signaling
and stomatal movement. Plant Cell 16, 1143–1162. http://dx.doi.org/10.1105/tpc.
021584.
de Witte, Y., Harkema, H., van Doorn, W.G., 2014. Effect of antimicrobial compounds on
cut Gerbera flowers: Poor relation between stem bending and numbers of bacteria in
the vase water. Postharvest Biol. Technol. 91, 78–83. http://dx.doi.org/10.1016/j.
postharvbio.2013.12.018.
Doi, M., Miyagawa-Namao, M., Inamoto, K., Imanishi, H., 1999. Rhythmic changes in
water uptake, transpiration and water potential of cut roses as affected by photo-
periods. J. Jpn. Soc. Hort. Sci. 68, 861–867. http://dx.doi.org/10.2503/jjshs.68.861.
Elibox, W., Umaharan, P., 2008. Morphophysiological characteristics associated with vase
life of cut flowers of anthurium. HortScience 43, 825–831 ISSN: 0018-5345.
Elibox, W., Umaharan, P., 2010. Cultivar differences in the deterioration of vase-life in
cut-flowers of Anthurium andraeanum is determined by mechanisms that regulate
water uptake. Sci. Hortic. 124, 102–108. http://dx.doi.org/10.1016/j.scienta.2009.
12.005.
Fanourakis, D., Carvalho, S.M., Almeida, D.P., van Kooten, O., van Doorn, W.G.,
Heuvelink, E., 2012. Postharvest water relations in cut rose cultivars with contrasting
sensitivity to high relative air humidity during growth. Postharvest Biol. Technol. 64,
X. Huang et al.
64–73. http://dx.doi.org/10.1016/j.postharvbio.2011.09.016.
Fanourakis, D., Heuvelink, E., Carvalho, S.M., 2013. A comprehensive analysis of the
physiological and anatomical components involved in higher water loss rates after
leaf development at high humidity. J. Plant Physiol. 170, 890–898. http://dx.doi.
org/10.1016/j.jplph.2013.01.013.
Fanourakis, D., Giday, H., Li, T., Kambourakis, E., Ligoxigakis, E.K., Papadimitriou, M.,
Strataridaki, A., Bouranis, D., Fiorani, F., Heuvelink, E., Ottosen, C.O., 2016.
Antitranspirant compounds alleviate the mild-desiccation-induced reduction of vase
life in cut roses. Postharvest Biol. Technol. 117, 110–117. http://dx.doi.org/10.
1016/j.postharvbio.2016.02.007.
Farrell, A.D., Evelyn, S., Lennon, A.M., Umaharan, P., 2012. Genotypic variation in se-
nescence and water relations in cut flowers of Anthurium andraeanum (Hort.).
HortScience 47, 1333–1337 ISSN: 0018-5345.
Frómeta, O.M., Morgado, M.M.E., da Silva, J.A.T., Morgado, D.T.P., Gradaille, M.A.D.,
2017. In vitro propagation of Gerbera jamesonii Bolus ex Hooker f. in a temporary
immersion bioreactor. Plant Cell Tiss. Org. 129, 543–551. http://dx.doi.org/10.
1007/s11240-017-1186-7.
Franks, P.J., Drake, P.L., Beerling, D.J., 2009. Plasticity in maximum stomatal con-
ductance constrained by negative correlation between stomatal size and density: an
analysis using Eucalyptus globulus. Plant Cell Environ. 32, 1737–1748. http://dx.doi.
org/10.1111/j.1365-3040.2009.002031.x.
Franks, P.J., Beerling, D.J., 2009. Maximum leaf conductance driven by CO2 effects on
stomatal size and density over geologic time. Proc. Natl. Acad. Sci. USA. 106,
10343–10347. http://dx.doi.org/10.1073/pnas.0904209106.
Halevy, A.H., Mayad, S., 1981. Senescence and postharvest physiology of cut flowers.
Hortic. Rev. 3, 59–143 ISSN: 0163-7851.
He, S.G., Joyce, D.C., Irving, D.E., Faragher, J.D., 2006. Stem end blockage in cut Grevillea
‘Crimson Yul-lo’ inflorescences. Postharvest Biol. Technol. 41, 78–84. http://dx.doi.
org/10.1016/j.postharvbio.2006.03.002.
Hetherington, A.M., Woodward, F.I., 2003. The role of stomata in sensing and driving
environmental change. Nature 424, 901–908. http://dx.doi.org/10.1038/
nature01843.
Hew, C.S., Lee, G.L., Wong, S.C., 1980. Occurrence of non-functional stomata in the
flowers of tropical orchids. Ann. Bot. 46, 195–201. http://dx.doi.org/10.1093/
oxfordjournals.aob.a085907.
In, B.C., Inamoto, K., Doi, M., Park, S.A., 2016. Using thermography to estimate leaf
transpiration rates in cut roses for the development of vase life prediction models.
Hortic. Environ. Biotechnol. 57, 53–60. http://dx.doi.org/10.1007/s13580-016-
0117-6.
In, B.C., Lim, J.H., 2018. Potential vase life of cut roses: Seasonal variation and re-
lationships with growth conditions, phenotypes, and gene expressions. Postharvest
Biol. Technol. 135, 93–103. http://dx.doi.org/10.1016/j.postharvbio.2017.09.006.
Kitamura, Y., Ueno, S., 2015. Inhibition of transpiration from the inflorescence extends
the vase life of cut hydrangea flowers. Hortic. J. 84, 156–160. http://dx.doi.org/10.
2503/hortj.MI-016.
Kohl, H.C., Rundle, D.L., 1972. Decreasing water loss off cut roses with abscisic acid.
HortScience 7 (249) ISSN: 0018-5345.
Kuang, Q., Li, L.F., Peng, J.Z., Sun, S.L., Wang, X.J., 2013. Transcriptome analysis of
Gerbera hybrida ray florets: putative genes associated with gibberellin metabolism
and signal transduction. PLoS One 8, e57715. http://dx.doi.org/10.1371/journal.
pone.0057715.
Liu, J.P., He, S.G., Zhang, Z.Q., Cao, J.P., Lv, P.T., He, S.D., Cheng, G.P., Joyce, D.C.,
2009. Nano-silver pulse treatments inhibit stem-end bacteria on cut gerbera cv.
Ruikou flowers. Postharvest Biol. Technol. 54, 37–40. http://dx.doi.org/10.1016/j.
postharvbio.2009.05.004.
Lü, P.T., Cao, J.P., He, S.G., Liu, J.P., Li, H.M., Cheng, G.P., Ding, Y.L., Joyce, D.C., 2010.
Nano-silver pulse treatments improve water relations of cut rose cv. Movie Star
flowers. Postharvest Biol. Technol. 57, 196–202. http://dx.doi.org/10.1016/j.
postharvbio.2010.04.003.
Lü, P.T., Huang, X.M., Li, H.M., Liu, J.P., He, S.G., Joyce, D.C., Zhang, Z.Q., 2011.
Continuous automatic measurement of water uptake and water loss of cut flower
stems. HortScience 46, 509–512 ISSN: 0018-5345.
45
Postharvest Biology and Technology 142 (2018) 39–45
Luomala, E., Laitinen, K., Sutinen, S., Kellomäki, S., Vapaavuori, E., 2005. Stomatal
density, anatomy and nutrient concentrations of scots pine needles are affected by
elevated CO2 and temperature. Plant Cell Environ. 28, 733–749. http://dx.doi.org/
10.1111/j.1365-3040.2005.01319.x.
Mattos, D.G., de Oliveira Paiva, P.D., Nery, F.C., Vale, R.P., Sarto, M.T., Luz, I.C.A., 2017.
Water relations in post-harvested torch ginger affected by harvest point and carnauba
wax. Postharvest Biol. Technol. 127, 35–43. http://dx.doi.org/10.1016/j.
postharvbio.2016.12.007.
Milla, R., de Diego-Vico, N., Martín-Robles, N., 2013. Shifts in stomatal traits following
the domestication of plant species. J. Exp. Bot. 64, 3137–3146. http://dx.doi.org/10.
1093/jxb/ert147.
Mori, I.C., Murata, Y., 2011. ABA signaling in stomatal guard cells: lessons from
Commelina and Vicia. J. Plant Res. 124, 477–487. http://dx.doi.org/10.1007/s10265-
011-0435-9.
Orcen, N., Nazarian, G., Gharibkhani, M., 2013. The responses of stomatal parameters
and SPAD value in Asian tobacco exposed to chromium. Pol. J. Environ. Stud. 22,
1441–1447 ISSN: 1230-1485.
Perik, R.R., Razé, D., Harkema, H., Zhong, Y., van Doorn, W.G., 2012. Bending in cut
Gerbera jamesonii flowers relates to adverse water relations and lack of stem scler-
enchyma development, not to expansion of the stem central cavity or stem elonga-
tion. Postharvest Biol. Technol. 74, 11–18. http://dx.doi.org/10.1016/j.postharvbio.
2012.06.009.
Perik, R.R., Razé, D., Ferrante, A., van Doorn, W.G., 2014. Stem bending in cut Gerbera
jamesonii flowers: Effects of a pulse treatment with sucrose and calcium ions.
Postharvest Biol. Technol. 98, 7–13. http://dx.doi.org/10.1016/j.postharvbio.2014.
06.008.
Pompelli, M.F., Martins, S.C.V., Celin, E.F., Ventrella, M.C., DaMatta, F.M., 2010. What is
the influence of ordinary epidermal cells and stomata on the leaf plasticity of coffee
plants grown under full-sun and shady conditions? Braz. J. Biol. 70, 1083–1088.
http://dx.doi.org/10.1590/S1519-69842010000500025.
Romero-Aranda, R., Cantó-Garay, R., Martínez, P.F., 1994. Distribution and density of
stomata in two cultivars of Gerbera jamesonii and its relation to leaf conductance. Sci.
Hortic. 58, 167–173. http://dx.doi.org/10.1016/0304-4238(94)90137-6.
Schroeder, K.R., Stimart, D.P., 2005. Comparison of stomatal density and postharvest
transpiration between long- and short-lived cut flower genotypes of Antirrhinum
majus L. J. Am. Soc. Hortic. Sci. 130, 243–244 ISSN: 0003-1062.
Slootweg, G., van Meeteren, U., 1991. Transpiration and stomatal conductance of roses
cv. Sonia grown with supplemental lighting. Acta. Hort. 298, 119–125. http://dx.doi.
org/10.17660/ActaHortic.1991.298.12.
Teeri, T.H., Uimari, A., Kotilainen, M., Laitinen, R., Help, H., Elomaa, P., Albert, V.A.,
2006. Reproductive meristem fates in Gerbera. J. Exp. Bot. 57, 3445–3455. http://dx.
doi.org/10.1093/jxb/erl181.
Torre, S., Fjeld, T., 2001. Water loss and postharvest characteristics of cut roses grown at
high or moderate relative air humidity. Sci. Hortic. 89, 217–226. http://dx.doi.org/
10.1016/S0304-4238(00)00229-6.
Torre, S., Fjeld, T., Gislerød, H.R., Moe, R., 2003. Leaf anatomy and stomatal morphology
of greenhouse roses grown at moderate or high air humidity. J. Am. Soc. Hortic. Sci.
128, 598–602 ISSN: 0003-1062.
van Doorn, W.G., 1997. Water relations of cut flowers. Hortic. Rev. 18, 1–85 ISSN: 0163-
7851.
van Doorn, W.G., 2012. Water relations of cut flowers: an update. Hortic. Rev. 40, 55.
http://dx.doi.org/10.1002/9781118351871.ch2.
van Meeteren, U., 1978. Water relations and keeping-quality of cut Gerbera flowers. II.
Water balance of ageing flowers. Sci. Hortic. 9, 189–197. http://dx.doi.org/10.1016/
0304-4238(78)90086-9.
Wang, W., Liu, Z., Bao, L.J., Zhang, S.S., Zhang, C.G., Li, X., Li, H.X., Zhang, X.L., Bones,
A.M., Yang, Z.B., Chen, Y.L., 2017. The Rho-GTPase RopGEF2-ROP7/ROP2 pathway
activated by phyB suppresses red light-induced stomatal opening. Plant Physiol. 174,
717–731. http://dx.doi.org/10.1104/pp.16.01727.
Zobayed, S.M.A., Armstrong, J., Armstrong, W., 2001. Leaf anatomy of in vitro tobacco
and cauliflower plantlets as affected by different types of ventilation. Plant Sci. 161,
537–548. http://dx.doi.org/10.1016/S0168-9452(01)00438-1.