Eco-Physiological Screening of Different Tomato Genotypes in Response to High Temperatures: A Combined Field-to-Laboratory Approach

Abstract and Figures

High temperatures represent a limitation for growth and development of many crop species. Several studies have demonstrated that the yield reduction of tomato under high temperatures and drought is mainly due to a photosynthetic decline. In this paper, a set of 15 tomato genotypes were screened for tolerance to elevated temperatures by cultivating plants under plastic walk-in tunnels. To assess the potential tolerance of tomato genotypes to high temperatures, measurements of chlorophyll fluorescence, pigments content and leaf functional traits have been carried out together with the evaluation of the final yields. Based on the greenhouse trials, a group of eight putative heat-sensitive and heat-tolerant tomato genotypes was selected for laboratory experiments aimed at investigating the effects of short-term high temperatures treatments in controlled conditions. The chlorophyll fluorescence induction kinetics were recorded on detached leaves treated for 60 min at 35 °C or at 45 °C. The last treatment significantly affected the photosystem II (PSII) photochemical efficiency (namely maximum PSII quantum efficiency, Fv/Fm, and quantum yield of PSII electron transport, ΦPSII) and the non-photochemical quenching (NPQ) in the majority of genotypes. The short-term heat shock treatments also led to significant differences in the shape of the slow Kautsky kinetics and its significant time points (chlorophyll fluorescence levels minimum O, peak P, semi-steady state S, maximum M, terminal steady state T) compared to the control, demonstrating heat shock-induced changes in PSII functionality. Genotypes potentially tolerant to high temperatures have been identified. Our findings support the idea that chlorophyll fluorescence parameters (i.e., ΦPSII or NPQ) and some leaf functional traits may be used as a tool to detect high temperatures-tolerant tomato cultivars.
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Plants 2020, 9, 508; doi:10.3390/plants9040508
Eco-Physiological Screening of Different Tomato
Genotypes in Response to High Temperatures: A
Combined Field-to-Laboratory Approach
Carmen Arena
, Stefano Conti
, Silvana Francesca
, Giuseppe Melchionna
, Josef Hájek
Miloš Barták
, Amalia Barone
and Maria Manuela Rigano
Department of Biology, University of Naples “Federico II”, Complesso Universitario Monte S. Angelo, Via
Cintia, 80126 Napoli, Italy;
Department of Agricultural Sciences, University of Naples “Federico II”, Via Università 100, 80055 Portici
(NA), Italy; (S.C.); (S.F.); (G.M.); (A.B.); (M.M.R.)
Department of Experimental Biology, Faculty of Science, Masaryk University, University Campus
Bohunice, Kamenice 753/5, 62500 Brno, Czech Republic; (J.H.); (M.B.)
* Correspondence:; Tel.: +0039-0812532125
Equally contributed.
Received: 31 March 2020; Accepted: 13 April 2020; Published: 15 April 2020
Abstract: High temperatures represent a limitation for growth and development of many crop
species. Several studies have demonstrated that the yield reduction of tomato under high
temperatures and drought is mainly due to a photosynthetic decline. In this paper, a set of 15 tomato
genotypes were screened for tolerance to elevated temperatures by cultivating plants under plastic
walk-in tunnels. To assess the potential tolerance of tomato genotypes to high temperatures,
measurements of chlorophyll fluorescence, pigments content and leaf functional traits have been
carried out together with the evaluation of the final yields. Based on the greenhouse trials, a group
of eight putative heat-sensitive and heat-tolerant tomato genotypes was selected for laboratory
experiments aimed at investigating the effects of short-term high temperatures treatments in
controlled conditions. The chlorophyll fluorescence induction kinetics were recorded on detached
leaves treated for 60 min at 35 °C or at 45 °C. The last treatment significantly affected the
photosystem II (PSII) photochemical efficiency (namely maximum PSII quantum efficiency, F
and quantum yield of PSII electron transport, Φ
) and the non-photochemical quenching (NPQ) in
the majority of genotypes. The short-term heat shock treatments also led to significant differences
in the shape of the slow Kautsky kinetics and its significant time points (chlorophyll fluorescence
levels minimum O, peak P, semi-steady state S, maximum M, terminal steady state T) compared to
the control, demonstrating heat shock-induced changes in PSII functionality. Genotypes potentially
tolerant to high temperatures have been identified. Our findings support the idea that chlorophyll
fluorescence parameters (i.e., Φ
or NPQ) and some leaf functional traits may be used as a tool to
detect high temperatures-tolerant tomato cultivars.
Keywords: heat stress; tomato genotypes; photosynthesis; crop yield; chlorophyll a fluorescence;
Solanum lycopersicum
1. Introduction
Increasing atmospheric temperatures, which are expected to rise by 2–4.8 °C in the next few
decades, can compromise crop productivity in numerous regions worldwide [1–4]. Indeed, elevated
temperatures can induce a series of physiological responses with consequent decreases of crops
Plants 2020, 9, 508 2 of 17
yields and quality [5,6]. Tomato (Solanum lycopersicum), being an excellent source of health-promoting
compounds, is one of the most important crops cultivated worldwide and its heat sensitivity varies
among different genotypes [4,7,8]. Generally, the optimal temperature range for photosynthesis is
considered to be between 25 °C and 30 °C [6]. The rising of average temperatures due to the ongoing
climate change will cause extensive productivity losses in Mediterranean areas, where tomato is
traditionally cultivated [9–11]. In this framework, it becomes important to perform studies that are
able to identify the most promising genotypes able to face heat stress.
The relationship between gas exchange and crop yield has been largely studied in tomato,
suggesting leaf transpiration as the most reliable indicator for yield prediction under drought [12].
However, beside gas exchange, other photosynthesis related parameters [13] should be taken into
account to build a “eco-physiological identity card” for different genotypes.
Chlorophyll fluorescence represents a good tool to rapidly and accurately detect plant health
status, the occurrence of damage within photosystem II (PSII), and to study heat tolerance in vivo
[6,13,14]. The decline of maximum quantum efficiency of PSII (F
), as well as the increase of non-
photochemical quenching (NPQ), are two heat-affected fluorescence parameters [15] related to
photoinhibition and photoprotection mechanisms in response to high temperatures [5,10,16]. Also,
transient changes in fluorescence intensity (Kautsky phenomenon) have been demonstrated to be
particularly suitable for the screening of physiological parameters in plants. The shape of
fluorescence curves changes significantly when plants switch from a healthy status to stress, giving
precious information on plant capability to overcome the stress [17].
Indeed, physiological screening techniques may complement phenotypic measurements and,
therefore, increase the efficiency of the selection of tolerant genotypes [18]. Currently, the majority of
the experiments on tomato responses to heat stress have been carried out in controlled chambers and
only few studies have been performed in the field [11,14,19].
On one hand, with the field approach, it is possible to screen a high number of different
genotypes. On the other hand, it is not easy to separate the effects of heat stress from other
environmental variables, such as light or water depletion, in inducing the plant specific responses
being examined [18]. For these reasons, field experiments should be complemented by laboratory
studies in which the use of novel screening methods, such as chlorophyll fluorescence imaging, may
provide further information for the characterization of tomato genotypes best suited to different
environmental conditions.
This study aimed to evaluate the responses to elevated temperatures in tomato genotypes in a
combined field-to-laboratory approach. First, we investigated the photosynthetic efficiency by
fluorescence emission measurements and leaf structural traits of different tomato varieties grown in
Mediterranean agro-ecosystems during summer in the field. These trials allowed us to identify
functional parameters correlating with final crop yields in different genotypes. After, a group of heat-
tolerant and heat-sensitive tomato genotypes were selected based on crop yield and photosynthesis-
related parameters. These selected genotypes were further characterized in the laboratory, analyzing
the Kautsky fluorescence induction curve after short-term heat treatments to obtain “a signature of
photosynthesis”. More specifically, the shapes of the curves changed when plants were subjected to
stress. The presence and the timing of the appearance of specific fluorescence transients (for
definitions, see [20]) were calculated in order to assess the heat stress-induced changes in PSII
functionality among cultivars.
These laboratory analyses allowed us to validate this easy and very quick protocol as an
alternative method for the selection of potentially heat-tolerant tomato genotypes. The combined
field-to-laboratory approach provides additional information which could help plant biologists and
breeders with characterizing responses to high temperatures in Solanum lycopersicum.
2. Results
2.1. Greenhouse Trials: Correlations Between Physiological Parameters and Yields
Plants 2020, 9, 508 3 of 17
Fifteen tomato genotypes, differing in geographical origin, were grown at supra-optimal
temperatures in order to identify the high and low performers in field environmental conditions. The
genotypes were selected on the basis of crop yield and eco-physiological indices. During anthesis,
maximum air temperatures inside the greenhouse were in the range of 24–43 °C. Significant
differences between each genotype vs. the control genotype JAG8810 were recorded for leaf
functional traits, chlorophylls and carotenoids contents and final yields, as reported in Table 1. The
hybrid JAG8810 had a good yield under high temperatures (from Monsanto, unpublished results).
Therefore, this line may be considered as a positive control which is able to resist extreme high
Table 1. Leaf traits (DW = dry weight of individual leaves, LA = leaf area, SLA = specific leaf area),
total carotenoids, chlorophylls (Chl a and Chl b) and crop yield per plant (YP) evaluated on tomato
genotypes grown under a plastic walk-in tunnel. Values are means ± standard deviation. ANOVA
with Least Significant Difference (LSD) post-hoc test was used to compare each genotype vs. the
control genotype JAG8810. The asterisks indicate statistically significant differences at * p < 0.05; ** p
< 0.01; *** p < 0.001.
(mg 100 g
Chl a (mg 100
Chl b (mg
100 g
(kg pt
E7 0.04 ± 0.10 11.41 ± 1.71 278.17 ± 63.62
* 28.76 ± 0.07 *** 104.82 ± 0.13
36.53 ± 0.29
0.98 ± 0.00
E8 0.10 ± 0.04 14.37 ± 2.86 155.73 ± 36.29 41.30 ± 0.25 *** 169.30 ± 0.27
76.05 ± 0.46
0.77 ± 0.17
E17 0.08 ± 0.02 17.76 ± 2.37 231.60 ± 45.53 44.67 ± 0.23 *** 177.92 ± 0.29
75.18 ± 0.77
1.26 ± 0.17
E36 0.14 ± 0.03
*** 20.09 ± 3.53 ** 146.83 ± 15.50 39.94 ± 0.10 *** 159.32 ± 0.37
67.97 ± 0.17
0.94 ± 0.22
0.15 ± 0.04
*** 13.12 ± 2.52
92.48 ±
17.82** 34.40 ± 0.03 ***
133.61 ± 0.64
54.61 ± 0.64
0.88 ± 0.17
E42 0.10 ± 0.03 18.02 ± 3.11 188.57 ± 37.70 43.02 ± 0.54 *** 174.35 ± 1.62
76.43 ± 0.83
*** 2.25 ± 1.04
E45 0.16 ± 0.02
27.58 ± 4.78
*** 171.16 ± 26.94 36.61 ± 0.04 * 142.81 ± 0.36
*** 59.02 ± 0.35 ** 2.24 ± 1.04
E53 0.07 ± 0.02 12.50 ± 2.19 210.79 ± 62.56 34.37 ± 0.06 *** 132.38 ± 0.10
53.01 ± 0.26
1.32 ± 0.28
E76 0.11 ± 0.01**
21.17 ± 2.25 ** 210.79 ± 62.56 44.63 ± 0.12 ***
171.90 ± 0.06
68.26 ± 0.37
0.64 ± 0.01
E107 0.07 ± 0.01 13.87 ± 2.95 194.36 ± 30.79 36.43 ± 0.04 ** 134.20 ± 0.28
48.49 ± 0.44
0.42 ± 0.12
IL12-4-SL 0.10 ± 0.02 21.00 ± 1.83 ** 217.16 ± 48.16 39.71 ± 0.07 *** 149.58 ± 0.11
56.68 ± 0.20
2.77 ± 0.24
JAG8810 0.07 ± 0.01 13.46 ± 1.86 203.15 ± 55.39 42.96 ± 0.29 161.63 ± 1.51 60.49 ± 1.25 2.99 ± 0.5
LA2662 0.12 ± 0.01
** 21.59 ± 2.07 ** 190.13 ± 24.57 36.98 ± 0.03 *** 143.90 ± 0.42
*** 58.78 ± 0.47 ** 1.97 ± 0.66
LA3120 0.13 ± 0.03 * 20.57 ± 1.14
*** 166.53 ± 36.45 39.30 ± 0.13 *** 155.43 ± 0.23
65.78 ± 0.60
*** 1.91 ± 0.98
M82 0.15 ± 0.03
24.37 ± 4.93
*** 163.26 ± 12.14 32.87 ± 0.14 *** 121.82 ± 0.23
44.88 ± 0.20
*** 3.25 ± 0.6 *
Leaf dry weight (DW) and leaf area (LA) of genotypes E36, E45, E76, LA2662, LA3120 and M82
were significantly higher compared to JAG8810. The genotype E7 was the only one with SLA values
higher than JAG8810. The pigments content (chlorophyll a and b and carotenoids) of all genotypes
was significantly lower than JAG8810, whereas only the M82 genotype showed a crop yield higher
than JAG8810, considered as the control. Chlorophyll fluorescence parameters (F
, Φ
and NPQ)
are shown in Figure 1.
Plants 2020, 9, 508 4 of 17
Figure 1. Maximum quantum yield of photosystem II (PSII) (F
), effective quantum yield of PSII
), and non-photochemical quenching (NPQ) measured on different tomato genotypes grown
under a plastic walk-in tunnel. Bars are means ± standard error (n = 5). ANOVA with LSD post-hoc
test was used to compare each genotype vs. the control genotype JAG8810 (green column). The
asterisks indicate statistically significant differences at * p < 0.05; ** p < 0.01; *** p < 0.001.
Based on field screening, the photochemical parameters differed among genotypes; more
specifically, seven genotypes, namely E8, E17, E32, E37, E42, IL12-4-SL and LA2662, showed higher
values compared to JAG8810. The genotypes E8, E42 and LA2662 also exhibited higher Φ
compared to the control genotype. Conversely, E76 and E107 genotypes showed the highest NPQ
values compared to the control and the other genotypes. The correlations among the analyzed
physiological and structural parameters are reported in Table 2.
Plants 2020, 9, 508 5 of 17
Table 2. Pearson’s correlations between physiological parameters, pigment content and crop yield
production (* p < 0.05; ** p < 0.01). YP = yield per plant; F
= maximum quantum yield of PSII; Φ
= effective quantum yield of PSII; NPQ = non-photochemical quenching; DW = leaf dry weight; SLA
= specific leaf area; LA = leaf area; Chl a = chlorophyll a; Chl b = chlorophyll b; Car = carotenoids.
Chl a
Chl b
−0.647 **
−0.650 **
0.744 **
Chl a
0.970 **
0.983 **
Chl b
0.909 **
Crop yield was negatively correlated with NPQ. Leaf area was positively correlated with leaf
DW. The maximum quantum efficiency of PSII (F
) differed among genotypes, although no
significant correlation was found between crop yield and F
. No correlations were found between
crop yield and chlorophylls or carotenoids content.
Based on crop yield (YP) and NPQ (higher YP and lower NPQ, Table 1 and Figure 1), the top
five performers (genotypes IL12-4-SL, JAG8810, LA3120, LA2662 and M82) along with the low
performer (genotype E107), were chosen for further analyses.
2.2. Chlorophyll a Fluorescence Measurements on Detached Leaves: Heat Shock Treatment at 35 °C and 45 °C
Chlorophyll fluorescence transients and derived parameters were measured as described in the
Materials and Methods section on detached leaves from the selected genotypes and from an
additional two tomato varieties, BG1620 and E41, which are supposedly heat-sensitive from previous
studies carried out in our laboratory.
No significant differences in the chlorophyll fluorescence parameters after a 60-min heat shock
at 35 °C were found between control and treated leaves (data not shown).
Conversely, heat shock at 45 °C for 60 min resulted in significant damage to PSII compared to
control (Figure 2).
Plants 2020, 9, 508 6 of 17
Figure 2. Maximum quantum yield of PSII (F
), effective quantum yield of PSII (Φ
) and non-
photochemical quenching (NPQ) in detached tomato leaves of different tomato genotypes following
short term heat treatment for 60 min at 45 °C (solid bars), compared with the respective non-treated
control (open bars). Bars are means ± standard error (n = 5). The asterisks indicate statistically
significant differences (* p < 0.05; ** p < 0.01; *** p < 0.001) according to Student’s t-test.
The short-term heat shock treatment led to a reduction in maximum PSII quantum efficiency
) in almost all genotypes. Interestingly, the highest reductions in F
were recorded in the
two supposedly heat-sensitive genotypes, BG1620 (−52%) and E41 (−19%), that were more affected
by heat stress compared to the genotype LA3120 (−12% F
reduction). Contrastingly, the F
ratio was little or not affected by the heat shock treatments in M82 (−4%) and LA2662 (0%) genotypes,
which were among the top performers in the field trial. The quantum efficiency of PSII electron
) was also affected by the heat shock treatment. For most genotypes, a significant
reduction in the Φ
was recorded and LA3120, BG1620, and E41 were found to be the most sensitive
genotypes with a decrease of −46%, −45% and −42% compared to the control, respectively. The M82
and IL12-4-SL genotypes were little or not affected by the heat shock treatment. As a consequence of
Plants 2020, 9, 508 7 of 17
the heat shock treatment, the NPQ values increased significantly in most genotypes compared to their
respective controls, but not in BG1620, IL12-4-SL and M82.
2.3. Heat-Induced Changes in Shape of Kautsky Kinetics
The shape of slow Kautsky kinetics and the derived parameters clearly showed that the effects
of heat treatment vary with genotype. As an example, in Figure 3, the slow Kautsky kinetics of the
least heat sensitive (IL12-4-SL) and the most heat sensitive (E107) genotype are reported. Heat
treatment leads to a reduction of the P peak in both genotypes compared to non-stressed controls.
This decline appears more pronounced for the heat sensitive genotype E107. The comparison among
genotypes showed that the M and S chlorophyll fluorescence signals were missing in heat-treated
samples and resulted in the absence of P/S, P/M, and S/M ratios compared to respective controls
(Table 3).
Figure 3. The effect of heat shock (60 min at 45 °C) on the shape of slow Kautsky kinetics in tomato
plants. Non-treated controls are indicated by open symbols (), heat shock-treated leaves by solid
symbols (). Less sensitive (IL12-4-SL) and substantially sensitive genotypes (E107) are presented.
The curves are means of at least 4 replicates (leaves). Data are normalized to background chlorophyll
fluorescence (F
). The chlorophyll fluorescence levels: O, fluorescence minimum; P, fluorescence peak;
S, semi-steady state; M, fluorescence maximum; T, terminal steady state are indicated.
Plants 2020, 9, 508 8 of 17
Table 3. Heat-induced changes in chlorophyll fluorescence parameters characterizing the shape of slow
Kautsky kinetics recorded for 8 tomato genotypes. Values are means of 5 replicates. Standard deviation
(not shown here) was under 4% of means. Asterisks indicate the statistical significance of the difference in
heat treated leaves compared to their respective non-treated control (* 0 to 50%; ** 50 to 100%; *** over
100%). O (origin) = minimum fluorescence level (also termed F
); P = peak fluorescence level reached after
1–2 s of actinic light exposure; S = semi-steady state of fluorescence emission; M = maximum of fluorescence;
T = terminal steady state chlorophyll fluorescence of the slow Kautsky kinetics; Fp = fluorescence peak; Fs
= fluorescence steady state; Rfd = relative fluorescence decline (vitality index); t
= time at which P
fluorescence peak is reached; t
= time at which S fluorescence level is reached; t
= time at which M
fluorescence level is reached; t
= time at which T fluorescence level is reached, Dip = decrease.
BG1620 E41 E107 IL12-4-SL JAG8810 LA2662 LA3120 M82
O/P 0.229 0.242 0.212 0.220 0.201 0.239 0.194 0.207
P/S 1.810 1.167 1.065 1.517 1.405 1.242 1.193 1.673
S/M 0.922 - 0.994 0.916 0.938 - 0.975 0.888
P/T (steady state) 2.665 3.101 2.101 2.482 2.428 2.887 2.694 2.801
M/T (steady state)
Rfd = (Fp−Fs)/Fs 1.665 2.101 1.101 1.482 1.428 1.887 1.694 1.801
6.080 3.360 4.080 4.080 4.080 4.080 −29.940 6.080
10.080 - 2.720 10.080 8.080 - 6.080 12.080
270.060 270.060 270.060 270.060 270.060 270.060 270.060 270.060
Dip at 26.080 - 52.080 42.080 - 48.080 86.080 38.080
Heat Treated (60 min at 45 °C)
0.713 ***
0.483 **
0.341 **
0.344 **
0.366 **
0.252 *
0.379 **
0.316 **
1.280 *
0.936 *
0.855 *
P/M - - - 1.166 * 0.929 * - - 0.853 *
S/M - - - 0.911 * 0.993 * - - 0.997 *
P/T (steady state) 1.658 * 2.729 * 2.680 * 2.250 * 2.407 * 2.779 * 2.881 * 2.302 *
M/T (steady state) - - - 1.929 * 2.591 * - - 2.700 *
Rfd = (Fp-Fs)/Fs
0.658 **
1.729 *
1.680 **
1.250 *
1.407 *
1.779 *
1.881 *
1.302 *
1.280 * 1.280 * 1.760 ** 1.120 * 1.280 * 1.040 1.280 1.040
- - - 4.080 2.240 *** - - 3.360
- - - 8.080 2.400 *** - - 4.080
255.060 * 270.060 * 245.060 * 260.060 * 245.060 * 260.060 * 270.060 * 250.06 *
Dip at
34.080 *
40.08 *
It may be generalized that for all tomato genotypes the shape of the slow Kautsky kinetics was
affected mainly during the early phase (i.e., within the first 60 sec after the actinic light was switched
on). The most pronounced change was found in the O/P ratio in BG1620 which showed a relative
change of about 200%, while other genotypes showed a much lower variation (LA2662: 5.4%). Apart
from the shape of the slow Kautsky kinetics and ratio values, the time at which the P, S, and M points
were reached differed within treatments. Heat treatment led to an increase in t
in four cultivars.
However, t
showed no change in three genotypes, and a decrease in the IL12-4-SL genotype.
3. Discussion
To date, there is a lack of knowledge about how differently sensitive/tolerant tomato genotypes
would respond to heat events. In heat-sensitive tomato genotypes, high temperatures are responsible
for the decrease in photosynthesis and overall crop yield [6].
In this paper, a set of eco-physiological parameters have been proposed to screen the most
promising tomato genotypes to be cultivated under elevated temperatures. A combined
field/laboratory experimental approach was performed. Firstly, a field trial was carried out under a
plastic walk-in tunnel to assess the field performances of tomato genotypes cultivated at high
temperatures (up to 43 °C) in terms of crop yield and physiological traits. In the second part of this
study, the heat-resistant and heat-sensitive genotypes were tested in the laboratory to analyze their
Plants 2020, 9, 508 9 of 17
responses to short-term heat shock and to investigate the photochemical behavior related to their
different field performance.
We suppose that at the temperatures considered in this study, the photosynthetic apparatus was
not damaged but rather regulated in the different genotypes, contributing to the degree of their
sensitivity or resistance to heat.
Our field studies indicated that tomato genotypes with higher yields also had lower NPQ values.
Non-photochemical quenching is considered a key mechanism in photoprotection against light and
temperature stress in higher plants [16,21]. High NPQ is often associated with conformational
changes within PSII, which transiently depresses CO
fixation [22].
In our trials, the top performers in terms of yield (genotypes IL12-4-SL, M82, LA2662, LA3120
and JAG8810) also showed the lowest NPQ values. Our data suggest that under experimental field
conditions, these tomato genotypes are more efficient in transferring the light excitation energy to
fixation, thus producing a higher photosynthetic carbon gain. Contrastingly, the highest NPQ
values measured in the low performing genotypes (E37, E76 and E107) correspond to more intense
thermal energy dissipation, leading to lower CO
assimilation and crop yield.
In heat-tolerant tomato genotypes, the NPQ protection is likely activated less promptly than in
the heat-sensitive genotypes. Conversely to NPQ, the F
ratio and Φ
did not show any
correlation with crop productivity, suggesting that the higher F
values in some genotypes may
indicate a better photosynthetic performance that is not always related to a higher crop yield. These
data are in contrast with findings of many authors, who demonstrated that the F
ratio is one of
the fluorescence parameters most affected by high temperatures and used it as an index for screening
tomato genotypes under heat stress [14,15,23]. Indeed, our data indicated that the high temperatures
experienced by plants in the field did not compromise the quantum yield of PSII in any genotype.
Based on the results of Hückstädt et al. [24], it may be hypothesized that the detrimental effect
of the high diurnal temperatures on photosystems has been compensated for by the optimal
temperatures [6] reached in the greenhouse during the night (see Figure 4).
Elevated temperatures could also determine a loss in the amount of photosynthetic pigments.
Therefore, the capacity of some genotypes to maintain higher pigments content under heat stress, as
well as to adjust some leaf functional traits (i.e., SLA, LA, RWC), are considered key heat tolerance-
linked traits [6,25–27].
For example, a higher SLA can be essential to obtain a higher potential evaporative demand and
a more extensive foliar display to capture more light [26]. However, in this work, no significant
correlation was found between final crop yield and pigments content or final crop yield and SLA,
indicating that these traits are not automatically linked to crop productivity. Therefore, these traits
were not considered for the selection of the heat-tolerant and heat-sensitive genotypes in this study.
On the basis of field trials, the tomato genotypes IL12-4-SL, JAG 8810, LA3120, LA2662 and M82
were selected as the top performers in terms of yield and low NPQ values, whereas the genotype
E107 was selected as a low performer considering the low photochemical efficiency and crop yield.
The heat sensitive BG1620 and E41 genotypes were added to the list of genotypes to be further studied
in the laboratory. For these tomato genotypes, the chlorophyll fluorescence transient analysis (slow
Kautsky kinetics) was performed in response to heat treatments to separate the effects of the high
temperature from other environmental constraints in the field.
In a previous study on tomato, plants exposed to 42 °C for 24 h showed a decline of net
photosynthesis, maximum PSII photochemical efficiency and electron transport rate [28]. Consistent
with these findings, our results further demonstrated that a short-term (60 min) heat shock at 45 °C
was sufficient to cause significant effects on the photochemistry in detached tomato leaves.
Interestingly, the F
ratio was found to be most affected in the heat-sensitive genotype BG1620
and in the low performer E107 genotype. An F
reduction of only 8% was registered in the
genotype LA2662, which was selected as heat-tolerant in the field experiment. These data are also in
agreement with Sharma et al. [18] who measured a reduction of F
in detached wheat leaves at 45
°C and Camejo et al. [29] who reported an F
decrease in a heat-susceptible tomato cultivar and
no changes in heat-tolerant cultivar.
Plants 2020, 9, 508 10 of 17
We supposed that the F
and PSII decreases in the sensitive (BG1620 and E41) genotypes
could be due to heat-induced structural modifications in PSII, particularly D1 protein oxidative
degradation. It has been demonstrated that heat stress may cause cleavage of the reaction center-
binding protein D1 and induce dissociation of a manganese-stabilizing 33 kDa protein from the PSII
reaction center complex [30]. Such oxidative damages have a strong positive relationship with the
accumulated levels of reactive oxygen species (ROS) and lipid peroxidation under heat stress [31].
However, it cannot be excluded that the significant reduction of F
observed in some
genotypes may also be associated with photoprotection mechanisms, as indicated by NPQ values
that peaked in correspondence to the reduction in F
. Simultaneously, protective mechanisms are
activated to protect the D1 protein, such as the expression of heat shock proteins (HSPs) like HSP21
that directly binds D1 to shield it against damage [32]. However, this does not seem to be the case for
the genotype BG1620, which was the most affected by the severe decline of photochemical and non-
photochemical processes.
The decrease in F
values in detached tomato leaves that were heat-treated at 40 °C was also
reported by Willits and Peet [33]. In our work, a heat shock treatment at 35 °C on detached leaves
was similarly tested with no significant effects on the photochemical efficiency of PSII (data not
shown), supporting the idea that such responses depend on the severity of the heat stress applied
The analysis of the shape of the slow Kautsky kinetics and the parameters calculated from its
significant time points (chlorophyll fluorescence levels O, P, S, M, T) revealed that heat shock
treatment led to significant differences compared with the control. Polyphasic changes in chlorophyll
fluorescence signal in the PSMT part of the Kautsky kinetics represent the combined effect of
photochemical and non-photochemical processes taking place in the chloroplast [34]. As the main
changes happened during the first 60 s of actinic light exposure, they might be attributed to the
interperiod of balancing the rate of primary photochemical processes in PSII to the rate of CO
Since the S and M chlorophyll fluorescence levels were generally missing in heat-treated tomato
leaves, the processes responsible for S and M interstates (i.e., the processes regulating the Calvin–
Benson cycle of CO
fixation, such as limitations in NADP+, phosphate pool equilibration, and
transmembrane ΔpH formation [20]) were overwhelmed by a strong non-photochemical quenching
that was activated by the heat shock treatment. This may be a consequence of the heat-induced
thermal dissipation of absorbed light energy, such as state 1 to state 2 transitions causing preferential
excitation of PSI and structural changes in thylakoid membranes, as reported by Marutani et al. [35].
The activation of these protective mechanisms may, however, lead to PSI damage in high temperature
treated plants due to over-reduction of the acceptor side of PSI [13].
The changes observed during the transition from P to S phase of the Kautsky kinetics indicate
the actual proportion between the mechanisms involved in photochemical and non-photochemical
quenching [36]. Since the parameters derived from the slow Kautsky kinetics responded to heat
treatment, it might be concluded that they have a high potential in the evaluation of heat effects on
the chloroplast function of tomato, as shown for light stress and leaf age effects by Nesterenko et al.
[37]. Many studies support the idea that a sustained increase in leaf photosynthesis can also lead to
an increase in total biomass production [38].
Overall, in this work, several useful photosynthetic parameters were identified, which could be
essential to detect and describe high temperature-tolerant tomato cultivars. These parameters could
be used as an effective tool for the prompt identification of tomato genotypes tolerant to high
4. Materials and Methods
4.1. Plant Material and Growth Conditions
Fifteen genotypes have been tested in the field. Of these genotypes, eleven genotypes (marked
as E## in Table 4) were selected based on their different productivity (demonstrated in a previous
Plants 2020, 9, 508 11 of 17
experiment conducted in the Campania region in the year 2016 [39]). The genotypes JAG8810
(Monsanto F
hybrid), LA2662 (Saladette) and LA3120 (Malintka) were reported to have high fruit
productivity under high temperatures (JAG8810, from Monsanto, unpublished results; LA2662 and
LA3120, Tomato Genetic Resources Center [40]). The JAG8810 hybrid may be considered as a positive
control which is able to resist extreme high temperatures. The M82 and the IL12-4-SL genotypes,
previously selected and characterized in a recent paper [41], were added to the list of genotypes to be
tested because their physiological response to elevated temperatures was unknown. One additional
tomato variety, BG1620 (kindly provided by Prof. G. Pevicharova, MVRCI Bulgary), was also added
to the set of genotypes to be analyzed. The genotypes BG1620 and E41 are supposedly heat-sensitive
based on previous analyses carried out in our laboratories (unpublished data).
Table 4. Tomato genotypes analyzed in this study.
No. Genotype Origin Common Name
1 E7 Italy Corbarino PC04
Corbarino PC05
3 E17 Italy Pantano Romanesco
4 E36 Italy Riccia San Vito
5 E37 Italy Siccagno
7 E42 Italy PI15250
8 E45 Italy SM246
9 E53 South America Latin American cultivar (Honduras)
Black Plum
11 E107 Europe E-L-19, Spain
12 JAG8810 - Monsanto F1 hybrid
13 M82 California M82
15 LA2662 - Saladette
16 LA3120 - Malintka
17 BG1620 Bulgary -
Tomato plants were grown in the year 2017 in Battipaglia (Salerno, Italy) (40°23′03 N, 17°21′17
E, 72 m a.s.l.) in a Mediterranean or Csa climate according to the Köppen classification scheme [42],
under walk-in thermal polyethylene tunnels. During the whole cultural cycle, climatic data were
recorded using the weather station VantagePro2 from Davis Instrument Corp. The maximum and
minimum temperatures during anthesis are reported in Figure 4. Spatial variation in temperature
within the walk-in tunnels was found to be minimal.
Plants 2020, 9, 508 12 of 17
Figure 4. Relative humidity (R.H.; open symbols, right axis) and maximum and minimum air
temperatures (solid symbols, left axis) during May–July 2017 inside a greenhouse in the experimental
field at Battipaglia (Salerno, Campania region, Italy).
The seeds of all genotypes were first rinsed and soaked in distilled water and then kept for 4
days in 8.5 cm diameter Petri dishes over 3 layers of filter paper saturated with distilled water. After
germination, the seeds were sown in seed trays kept in the greenhouse. Seedlings were transplanted
in April under plastic walk-in tunnels. Plants were grown following the standard cultural practices
of the area. Insecticides and fungicides were applied to the plants according to general local practices
and recommendations. Urea phosphate fertilizer (40 kg ha
) was applied to the soil before
transplanting. Tillage treatments included plowing that was followed by one/two milling. Weeding
and ridging were also carried out. Through fertirrigation, recommended levels of N (190 kg ha
) and
K (20 kg ha
) were applied. During cultivation, plants were irrigated as required.
All genotypes were grown according to a completely randomized experimental design with
three replicates and 10 plants per replicate. Total fruit number and fresh weight were measured at the
end of growth season to evaluate the crop yield per plant (YP). The crop yield was measured at the
red fruit ripe stage.
4.2. Functional Leaf Trait Analysis
The measurements of leaf area (LA), specific leaf area (SLA) and leaf dry weight (DW) were
performed on the fourth leaf from the apex in each plant. Five leaves for each genotype were sampled
from 5 different plants. LA was measured using ImageJ 1.45 software for image analysis [43]. The
leaves were then dried at 70 °C and their DW was measured after 48 h. SLA was calculated as the
ratio of leaf area to leaf dry weight and expressed as cm
DW according to Cornelissen et al. [44].
4.3. Chlorophyll Fluorescence Emission Measurements in the Field
Chlorophyll fluorescence parameters were measured on fully expanded leaves (the fourth leaf
from the apex) using a portable FluorPen FP100 Max fluorometer, equipped with a
Photosynthetically Active Radiation (PAR) sensor (Photon Systems Instruments, Drásov, Czech
Republic) following the procedure reported by Sorrentino et al. [45]. Five replicate measurements for
each genotype were taken as follows.
The ground state fluorescence (F
) was induced by an internal Light Emitting Diode (LED) blue
(1–2 μmol photons m
) on 30-min dark-adapted leaves of plants moved into a dark room. The
maximum fluorescence level in the dark-adapted state (F
), was triggered by a 1 s saturating light
pulse of 3000 μmol photons m
, and the maximum quantum efficiency of PSII (F
) was
calculated according to Equation (1).
The fluorescence readings in the light were taken using an open leaf-clip, allowing for
measurements of the steady state fluorescence level (Fs) at an ambient light Photosynthetic Photon
Flux Density (PPFD) of 150–200 μmol m
in the PAR spectrum and of the maximum fluorescence
level in light-adapted leaves (F’
) measured after a saturating light pulse.
The quantum yield of PSII electron transport
) was calculated according to Equation (2)
[46]. Non-photochemical quenching (NPQ) was calculated using Equation (3) [47].
= (F
− F
= (F’
− F
NPQ = (F
− F’
4.4. Determination of Total Chlorophylls and Carotenoids Content
Following the chlorophyll fluorescence field measurements, the same leaves were excised,
stored in a cool box and transferred to the laboratory for the determination of photosynthetic
pigments content (chlorophyll a, chlorophyll b and carotenoids) according to the method described
Plants 2020, 9, 508 13 of 17
by Rigano et al. [5]. One gram of leaf sample was extracted with 16 mL of acetone/hexane (40/60, v/v)
with a T-25 Ultra-Turrax Homogenizer (IKA-Werke GmbH & Co. KG, Staufen, Germany). The
homogenate was then centrifuged at 5000 rpm for 5 min at 4 °C and supernatants were collected and
stored at −20 °C prior to spectrophotometric analysis. Pigment contents were calculated in mg on 100
g of leaf fresh weight. Three separate biological replicates for each sample and three technical assays
for each biological repetition were measured.
4.5. Chlorophyll a Fluorescence—Heat Treatments and Laboratory Measurements
Chlorophyll fluorescence transients and numeric parameters were measured with a FluorCam
(Photon Systems Instruments, Brno, Czech Republic) on detached leaves from genotypes selected
from the in-field analyses. Two additional tomato varieties, BG1620 and E41 (supposedly heat-
sensitive based on previous analyses carried out in our laboratories (unpublished data), were also
added to the set of genotypes to be studied in this research.
Whole compound leaves from field grown tomato plants were sampled in the morning, between
2 and 3 h after dawn (8:00 to 9:00 a.m.). Fully expanded leaves (the fourth leaf from the apex) were
excised at the base of the petiole from each plant using a sharp blade and the cut base was
immediately immersed in distilled water in a 50 mL test tube in order to prevent dehydration. The
sampled leaves were then temporarily stored in a dark cool box and transferred to the laboratory for
the short-term high temperature treatments and chlorophyll fluorescence experiments.
In the laboratory, single leaflets were excised from the tomato compound leaves and placed in
9.0 cm diameter Petri dishes over water saturated filter paper. Two groups of leaf samples were
selected for each genotype: one group was kept at the laboratory room temperature (25–26°C) in the
dark and assumed as control treatment, while another group was placed in a thermostatic cabinet
set, either at 35°C or at 45°C, for 1 h in the dark and was considered the heat treatment. At the end of
the heat treatment, leaf samples were adapted at room temperature for 20 min in the dark prior to
performing the fluorescence measurements. The same procedure was replicated on different leaves
and repeated for each tomato genotype. The whole experiment was carried out subsequently on leaf
samples treated either at 35 °C (lower heat stress level) or at 45 °C (higher heat stress level), plus the
respective controls.
The temperature conditions in this experiment were chosen to represent the optimal
temperature range for tomato (within 25 – 30 °C [6]) and the maximum air temperature of 33–43 °C
encountered by tomato genotypes in the walk-in tunnels during the experimental period.
Chlorophyll fluorescence transients (slow Kautsky kinetics supplemented with quenching analysis)
were measured with a Handy Fluor Cam FC-1000H imaging fluorimeter (Photon Systems
Instruments, Drásov, Czech Republic) controlled by the FluorCam7 software (Photon Systems
Instruments, Drásov, Czech Republic).
The experimental protocol started with the measurement of the ground state (minimum)
fluorescence level (F
, O) when the samples were exposed to low intensity measuring light flashes,
followed by a saturating pulse of light (960 ms, 2400 μmol m
) to induce maximum chlorophyll
fluorescence (F
, P). After 27 s of dark adaptation, the samples were exposed to actinic light (200
μmol m
) for 5 min until steady state chlorophyll fluorescence (F
) was reached. At this point,
another saturating pulse of light induced the maximum chlorophyll fluorescence of the light-adapted
sample (F’
). The maximum quantum efficiency of PSII (F
), the quantum efficiency of PSII
electron transport
) and non-photochemical quenching (NPQ) were calculated using the
FluorCam7 software, according to Equations (1) – (3) reported above, respectively.
4.6. Analysis of Kautsky Kinetics Shape in Response to Heat Treatment
Slow Kautsky kinetics is routinely used to evaluate the sensitivity of plants to a wide variety of
stressors [48,49]. In this study, the analysis of kinetic fluorescence shape, i.e., the presence and the
time of appearance of specific fluorescent transients (O, P, S, M and T), was utilized to assess heat
stress-induced changes in PSII functionality in tomato. For individual Kautsky kinetics recorded after
a heat treatment (see above), chlorophyll fluorescence levels O, P, S, M and T were identified, as well
Plants 2020, 9, 508 14 of 17
as the times at which they were reached. Effects of experimental temperature on the O-, P-, S-, M-,
and T-derived parameters were then evaluated for the individual genotypes and the genotype-
dependent responses of the parameters were characterized.
4.7. Statistical Analysis
Statistical analysis was performed on all measured traits using SPSS 23 Software (IBM SPSS
Statistics, USA). Analysis of variance (ANOVA) was used to check for significant differences between
each genotype vs. the control genotype (JAG8810) and where significant differences were found, the
least significant difference (LSD) at the 0.05, 0.01 or 0.001 level of probability was calculated and used
to compare the mean values. Student’s t-test was performed to check for differences between control
and heat-treated samples in the case of detached leaf experiments. Pearson’s correlation coefficient
was used to test associations between tomato yield and other variables.
5. Conclusions
Due to ongoing climate change, the screening and identification of the most promising tomato
cultivars able to maintain elevated productivity under heat stress becomes a priority for farmers and
producers to avoid significant losses of crop yield.
In our experiments, heat tolerant and heat sensitive tomato cultivars were identified and
characterized using a correlative approach combining different field and laboratory methods based
on functional leaf traits, crop yield and photochemical indexes.
The three main outcomes emerging from this work include the confirmation that some
parameters linked to chlorophyll fluorescence emission can be used to phenotype heat tolerance in
tomato, both in the field and in the laboratory. Secondly, we demonstrated that the detached leaf
method can be used as an easy, quick and valid alternative for the selection and characterization of
potentially heat-tolerant tomato genotypes. The advantage of a laboratory approach that implements
field measurements is that it is possible to separate the effects caused by heat treatments from the
other related environmental factors such as high light, low relative humidity and limited water
supply. Finally, we identified five tomato genotypes (JAG8810, LA3120, LA2662, IL12-4-SL and M82)
as promising genotypes that are potentially tolerant to elevated temperatures. These genotypes also
represent a valuable resource to be used in future works aiming to assess the underlying
physiological mechanisms for variability in photosynthetic responses among different cultivars.
Author Contributions: Conceptualization, C.A., S.C., A.B. and M.M.R.; Data curation, C.A., S.C., S.F., J.H., M.B.
and M.M.R.; Funding acquisition, A.B.; Investigation, C.A., S.C., S.F., G.M., J.H., M.B. and M.M.R.; Writing—
original draft, C.A., S.C., M.B., A.B. and M.M.R.; Writing—review and editing, C.A. and M.M.R. All authors
have read and agreed to the published version of the manuscript.
Funding: The authors have received funding from the European Union’s Horizon 2020 research and innovation
programme through the TomGEM project under grant agreement no. 679796.
Conflicts of Interest: The authors declare no conflict of interest.
1. IPCC. Climate Change 2013: The Physical Science Basis. Contribution of Working Group I to the Fifth Assessment
Report of the Intergovernmental Panel on Climate Change; Stocker, T.F., Qin, D., Plattner, G.-K., Tignor, M.M.B.,
Allen, S.K., Boschung, J., Nauels, A., Xia, W., Bex, V., Midgley, P.M. Eds.; Cambridge University Press:
Cambridge, UK.; New York, NY, USA, 2013.
2. Vitale, L.; Arena, C.; Carillo, P.; Di Tommasi, P.; Mesolella, B.; Nacca, F.; De Santo, A.; Fuggi, A.; Magliulo,
V. Gas exchange and leaf metabolism of irrigated maize at different growth stages. Plant. Biosyst. 2011, 145,
3. Sehgal, A.; Sita, K.; Siddique, K.H.M.; Kumar, R.; Bhogireddy, S.; Varshney, R.K.; Hanumanth Rao, B.; Nair,
R.M.; Prasad, P.V.V.; Nayyar, H. Drought or/and heat-stress effects on seed filling in food crops: Impacts
on functional biochemistry, seed yields, and nutritional quality. Front. Plant. Sci. 2018, 9, 1705,
Plants 2020, 9, 508 15 of 17
4. Zhou, R.; Wu, Z.; Wang, X.; Rosenqvist, E.; Wang, Y.; Zhao, T.; Ottosen, C.O. Evaluation of temperature
stress tolerance in cultivated and wild tomatoes using photosynthesis and chlorophyll fluorescence. Hortic.
Environ. Biotechnol. 2018, 59, 499–509.
5. Rigano, M.M.; Arena, C.; Di Matteo, A.; Sellitto, S.; Frusciante, L.; Barone, A. Eco-physiological response to
water stress of drought-tolerant and drought-sensitive tomato genotypes. Plant. Biosyst. 2016, 150, 682–691.
6. Zhou, R.; Kjaer, K.H.; Rosenqvist, E.; Yu, X.; Wu, Z.; Ottosen, C.O. Physiological response to heat stress
during seedling and anthesis stage in tomato genotypes differing in heat tolerance. J. Agron. Crop. Sci. 2016,
203, 68–80.
7. Francesca, S.; Arena, C.; Hay-Mele, B.; Schettini, C.; Ambrosino, P.; Barone, A.; Rigano, M.M. The use of a
plant-based biostimulant improves plant performances and fruit quality in tomato plants grown at elevated
temperatures. Agronomy 2020, 10, 363.
8. Zhou, R.; Yu, X.; Ottosen, C.O.; Rosenqvist, E.; Zhao, L.; Wang, Y.; Yu, W.; Zhao, T.; Wu, Z. Drought stress
had a predominant effect overheat stress on three tomato cultivars subjected to combined stress. BMC Plant.
Biol. 2017, 17, 24, doi:10.1186/s12870-017-0974-x.
9. Mathur, S.; Agrawal, D.; Jajoo, A. Photosynthesis: Responses to high temperature stress. J. Photochem.
Photobiol. B 2014, 137, 116–126.
10. Carvalho, L.C.; Coito, J.L.; Goncalves, E.F.; Chaves, M.M.; Amancio, S. Differential physiological response
of the grapevine varieties Touriga Nacional and Tricadeira to combined heat, drought and light stresses.
Plant. Biol. 2016, 18, 101–111.
11. Giorio, P.; Guida, G.; Mistretta, C.; Sellami, H.M.; Oliva, M.; Punzo, P.; Iovieno, P.; Arena, C.; De Maio, A.;
Grillo, S.; et al. Physiological, biochemical and molecular responses to water stress and rehydration in
Mediterranean adapted tomato landraces. Plant. Biol. 2018, 20, 995–1004.
12. Patanè, C.; Scordia, D.; Testa, G.; Cosentino, L.S. Physiological screening for drought tolerance in
Mediterranean long-storage tomato. Plant. Sci. 2016, 249, 25–34.
13. Brestič, M.; Živčák, M.; Kunderliková, K.; Allakhverdiev, S.I. High temperature specifically affects the
photoprotective responses of chlorophyll b-deficient wheat mutant lines. Photosynth. Res. 2016, 130, 251–
14. Poudyal, D.; Rosenqvist, E.; Ottosen, C. Phenotyping from lab to field – tomato lines screened for heat stress
using Fv/Fm maintain high fruit yield during thermal stress in the field. Funct. Plant. Biol. 2019, 46, 44–55,
15. Zhou, R.; Yu, X.; Kjaer, K.H.; Rosenqvist, E.; Ottosen, C. Screening and validation of tomato genotypes
under heat stress using Fv/Fm to reveal the physiological mechanism of heat tolerance. Environ. Exp. Bot.
2015, 118, 1–11.
16. Arena, C.; Vitale, L.; Virzo De Santo, A. Photosynthesis and photoprotective strategies in Laurus nobilis L.
and Quercus ilex L. under summer drought and winter cold. Plant. Biosyst. 2008, 142, 472–479.
17. Tyystjärvi, E.; Koski, A.; Keränen, M.K.; Nevalainen, O. The Kautsky Curve Is a Built-in Barcode. Biophys.
J. 1999, 77, 1159–1167.
18. Sharma, D.K.; Fernandez, J.O.; Rosenqvist, E.; Ottosen, C.; Andersen, S.B. Genotypic response of detached
leaves versus intact plants for chlorophyll fluorescence parameters under high temperature stress in wheat.
J. Plant. Physiol. 2014, 171, 576–586.
19. Prasanth, V.V.; Babu, M.S.; Basava, R.K.; Tripura Venkata, V.G.N.; Mangrauthia, S.K.; Voleti, S.R.;
Neelamraju, S. Trait and marker associations in Oryza nivara and O. rufipogon derived rice lines under two
different heat stress conditions. Front. Plant. Sci. 2017, 8, 1819, doi:10.3389/fpls.2017.01819.
20. Stirbet, A.; Riznichenko, G.; Rubin, A.B. Modeling chlorophyll a fluorescence transient: Relation to
photosynthesis. Biochemistry (Moscow) 2014, 79, 291–323.
21. Vitale, L.; Arena, C.; Virzo De Santo, A.; D’Ambrosio, N. Effects of heat stress on gas exchange and
Photosystem II (PSII) photochemical activity of Phillyrea angustifolia exposed to elevated CO
subsaturating irradiance. Botany 2008, 86, 435–441.
22. Kromdijk, J.; Głowacka, K.; Leonelli, L.; Gabilly, S.T.; Iwai, M.; Niyogi, K.K.; Long, S.P. Improving
photosynthesis and crop productivity by accelerating recovery from photoprotection. Science 2016, 354,
23. Sharma, D.K.; Andersen, S.B.; Ottosen, C.O.; Rosenqvist, E. Wheat cultivars selected for high Fv/Fm under
heat stress maintain high photosynthesis, total chlorophyll, stomatal conductance, transpiration and dry
matter. Physiol. Plant. 2015, 153, 284–298.
Plants 2020, 9, 508 16 of 17
24. Hückstädt, A.B.; Suthaparan, A.; Mortensen, L.M.; Gislerød, H.R. The effect of low night and high day
temperatures on photosynthesis in tomato. Am. J. Plant. Sci. 2013, 4, 2323–2331.
25. Nankishore, A.; Farrell, A. The response of contrasting tomato genotypes to combined heat and drought
stress. J. Plant. Physiol. 2016, 202, 75–82, doi:10.1016/j.jplph.2016.07.006.
26. Wang, D.; Heckathorn, S.A.; Mainali, K.; Tripathee, R. Timing effects of heat-stress on plant
ecophysiological characteristics and growth. Front. Plant. Sci. 2016, 7, 1629.
27. Ahammed, G.J.; Xu, W.; Liu, A.; Chen, S. COMT1 silencing aggravates heat stress-induced reduction in
photosynthesis by decreasing chlorophyll content, photosystem II activity and electron transport efficiency
in tomato. Front. Plant. Sci. 2018, 9, 998.
28. Pan, C.; Ahammed, G.J.; Li, X.; Shi, K. Elevated CO
improves photosynthesis under high temperature by
attenuating the functional limitations to energy fluxes, electron transport and redox homeostasis in tomato
leaves. Front. Plant. Sci. 2018, 9, 1739, doi.10.3389/fpls.2018.01739.
29. Camejo, D.; Rodríguez, P.; Angeles Morales, M.; Dell’Amico, J.M.; Torrecillas, A.; Alarcón, J.J. High
temperature effects on photosynthetic activity of two tomato cultivars with different heat susceptibility. J.
Plant. Physiol. 2005, 162, 281–289.
30. Yamane, Y.; Kashino, Y.; Koike, H.; Satoh, K. Effects of high temperatures on the photosynthetic systems
in spinach: Oxygen-evolving activities, fluorescence characteristics and the denaturation process.
Photosynth. Res. 1998, 57, 51–59.
31. Yamashita, A.; Nijo, N.; Pospíšil, P.; Morita, N.; Takenaka, D.; Aminaka, R.; Yamamoto, Y. Quality control
of Photosystem II—Reactive oxygen species are responsible for the damage to Photosystem II under
moderate heat stress. J. Biol. Chem. 2008, 283, 28380–28391.
32. Wang, Q.L.; Chen, J.H.; He, N.Y.; Guo, F.Q. Metabolic reprogramming in chloroplasts under heat stress in
plants. Int. J. Mol. Sci. 2018, 19, 849, doi:10.3390/ijms19030849.
33. Willits, D.H.; Peet, M.M. Measurement of chlorophyll fluorescence as a heat stress indicator in tomato:
Laboratory and greenhouse comparisons. J. Am. Soc. Hortic. Sci. 2001, 126, 188–194.
34. Riznichenko, G.; Lebedeva, G.; Pogosyan, S.; Sivchenko, M.; Rubin, A. Fluorescence induction curves
registered from individual microalgae cenobiums in the process of population growth. Photosynt. Res. 1996,
49, 151–157.
35. Marutani, Y.; Yamauchi, Y.; Miyoshi, A.; Inoue, K.; Ikeda, K.; Mizutani, M.; Sugimoto, Y. Regulation of
photochemical energy transfer accompanied by structural changes in thylakoid membranes of heat-
stressed wheat. Int. J. Mol. Sci. 2014, 15, 23042–23058.
36. Papageorgiou, G.C. The non-photochemical quenching of the electronically excited state of chlorophyll a
in plants: Definitions, timelines, viewpoints, open questions. In Non-Photochemical Quenching and Energy
Dissipation in Plants, Algae and Cyanobacteria; Demmig, B.A., Garab, G., Adams, W., III, Eds.; Springer:
Dordrecht, The Netherlands; Heidelberg, Germany; New York, NY, USA.; London, UK; 2014; pp. 1–44.
37. Nesterenko, T.V.; Tikhomirov, A.A.; Shikhov, V.N. Influence of excitation light intensity and leaf age on
the slow chlorophyll fluorescence transient in radish. Biophysics 2012, 57, 464–468.
38. Brestič, M.; Živčák, M.; Hauptvogel, P.; Misheva, S.; Kocheva, K.; Yang, X.H.; Li, X.N.; Allakhverdiev, S.I.
Wheat plant selection for high yields entailed improvement of leaf anatomical and biochemical traits
including tolerance to non-optimal temperature conditions. Photosynth. Res. 2018, 136, 245–255.
39. Ruggieri, V.; Calafiore, R.; Schettini, C.; Rigano, M.M.; Olivieri, F.; Frusciante, L.; Barone, A. Exploiting
genetic and genomic resources to enhance heat-tolerance in tomatoes. Agronomy 2019, 9, 22,
40. Tomato Genetic Resources Center. Available online:
(accessed on 14 January 2020)
41. Rigano, M.M.; Lionetti, V.; Raiola, A.; Bellincampi, D.; Barone, A. Pectic enzymes as potential enhancers of
ascorbic acid production through the D-galacturonate pathway in Solanaceae. Plant. Sci. 2018, 266, 55–63.
42. Peel, M.C.; Finlayson, B.L.; McMahon, T.A. Updated world map of the Koppen–Geiger climate
classification. Hydrol. Earth Syst. Sci. 2007, 11, 1633–1716.
43. Schneider, C.A.; Rasband, W.S.; Eliceiri, K.W. NIH Image to ImageJ: 25 years of image analysis. Nat.
Methods 2012, 9, 671–675.
44. Cornelissen, J.H.C.; Lavorel, A.; Garnier, E.; Diaz, S.; Buchmann, N.; Gurvich, D.E.; Reich, P.B.; ter Steege,
H.; Morgan, H.D.; van der Heijden, M.G.A.; et al. Handbook of protocols for standardized and easy
measurement of plant functional traits worldwide. Aus. J. Bot. 2003, 51, 335–380.
Plants 2020, 9, 508 17 of 17
45. Sorrentino, M.C.; Capozzi, F.; Amitrano, C.; Giordano, S.; Arena, C.; Spagnuolo, V. Performance of three
cardoon cultivars in an industrial heavy metal-contaminated soil: Effects on morphology, cytology and
photosynthesis. J. Hazard. Mater. 2018, 351, 131–137.
46. Genty, B.; Briantais, J.M.; Baker, N.R. The relationship between the quantum yield of photosynthetic
electron transport and quenching of chlorophyll fluorescence. Biochim. Biophys. Acta 1989, 990, 87–92.
47. Bilger, W.; Björkman, O. Role of the xanthophyll cycle in photoprotection elucidated by measurements of
light-induced absorbance changes, fluorescence and photosynthesis in leaves of Hedera canariensis.
Photosynth. Res. 1990, 25, 173–185.
48. Baker, N.R.; Rosenqvist, E. Applications of chlorophyll fluorescence can improve crop production
strategies: An examination of future possibilities. J. Exp. Bot. 2004, 55, 1607–1621.
49. Roháček, K.; Soukupová, J.; Barták, M. Chlorophyll fluorescence: A wonderful tool to study plant
physiology and plant stress. In Plant Cell Compartments; Schoefs, B., Ed.; Research Signpost: Kerala, India,
2008; pp. 41–104.
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article distributed under the terms and conditions of the Creative Commons Attribution
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... Moreover, most of the experiments were conducted under controlled conditions in greenhouses. Physiological measurements, such as chlorophyll fluorescence, have been proposed as quick tools for screening heat tolerance in tomato [28][29][30]. In some cases, interesting and good correlations have been found between chlorophyll fluorescence measurements in controlled greenhouse and field performance studies under high temperatures [28,29], although these correlations have not been found with crop yield in all the studies [28]. ...
... Physiological measurements, such as chlorophyll fluorescence, have been proposed as quick tools for screening heat tolerance in tomato [28][29][30]. In some cases, interesting and good correlations have been found between chlorophyll fluorescence measurements in controlled greenhouse and field performance studies under high temperatures [28,29], although these correlations have not been found with crop yield in all the studies [28]. However, the number of tested accessions and trials is still too low to draw general conclusions. ...
... Physiological measurements, such as chlorophyll fluorescence, have been proposed as quick tools for screening heat tolerance in tomato [28][29][30]. In some cases, interesting and good correlations have been found between chlorophyll fluorescence measurements in controlled greenhouse and field performance studies under high temperatures [28,29], although these correlations have not been found with crop yield in all the studies [28]. However, the number of tested accessions and trials is still too low to draw general conclusions. ...
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Background Due to global warming, the search for new sources for heat tolerance and the identification of genes involved in this process has become an important challenge as of today. The main objective of the current research was to verify whether the heat tolerance determined in controlled greenhouse experiments could be a good predictor of the agronomic performance in field cultivation under climatic high temperature stress. Results Tomato accessions were grown in greenhouse under three temperature regimes: control (T1), moderate (T2) and extreme heat stress (T3). Reproductive traits (flower and fruit number and fruit set) were used to define heat tolerance. In a first screening, heat tolerance was evaluated in 219 tomato accessions. A total of 51 accessions were identified as being potentially heat tolerant. Among those, 28 accessions, together with 10 accessions from Italy (7) and Bulgaria (3), selected for their heat tolerance in the field in parallel experiments, were re-evaluated at three temperature treatments. Sixteen tomato accessions showed a significant heat tolerance at T3, including five wild species, two traditional cultivars and four commercial varieties, one accession from Bulgaria and four from Italy. The 15 most promising accessions for heat tolerance were assayed in field trials in Italy and Bulgaria, confirming the good performance of most of them at high temperatures. Finally, a differential gene expression analysis in pre-anthesis (ovary) and post-anthesis (developing fruit) under heat stress among pairs of contrasting genotypes (tolerant and sensitive from traditional and modern groups) showed that the major differential responses were produced in post-anthesis fruit. The response of the sensitive genotypes included the induction of HSP genes, whereas the tolerant genotype response included the induction of genes involved in the regulation of hormones or enzymes such as abscisic acid and transferases. Conclusions The high temperature tolerance of fifteen tomato accessions observed in controlled greenhouse experiments were confirmed in agronomic field experiments providing new sources of heat tolerance that could be incorporated into breeding programs. A DEG analysis showed the complex response of tomato to heat and deciphered the different mechanisms activated in sensitive and tolerant tomato accessions under heat stress.
... Plants are continuously subjected to many abiotic and biotic stresses, from seed germination throughout the whole life cycle (Deryng et al. 2014), which are now intensified by climate change. In particular, water and heat stress are two of the most critical abiotic stresses limiting crop growth and productivity worldwide, especially in arid and semi-arid areas (Vitale et al. 2009;Arena et al. 2020). In the past few years, the combination of high temperature and water scarcity has caused global losses in crop production id~US$30 billion (Gupta et al. 2020). ...
... Tomato (Solanum lycopersicum L.) is one of the most cultivated vegetable crops worldwide. It has an optimal growth diurnal temperature range of 25-30°C and is known to be sensitive to both water shortage and heat, although its sensitivity varies among genotypes (Rigano et al. 2016;Zhou et al. 2018b;2020b;Arena et al. 2020;Francesca et al. 2020). Climate change is exacerbating and/or increasing the frequency of high temperature shocks and water shortages in the Mediterranean Basin, pointing to the need to develop varieties with enhanced tolerance to naturally co-occurring stresses (Zhou et al. 2020a,b). ...
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• Climate change is increasing the frequency of high temperature shocks and water shortages, pointing to the need to develop novel tolerant varieties and to understand the mechanisms employed to withstand combined abiotic stresses. • Two tomato genotypes, a heat-tolerant Solanum lycopersicum accession (LA3120) and a novel genotype (E42), previously selected as a stable yielding genotype under high temperatures, were exposed to single and combined water and heat stress. Plant functional traits, pollen viability and physiological (leaf gas exchange and chlorophyll a fluorescence emission measurements) and biochemical (antioxidant content and antioxidant enzyme activity) measurements were carried out. A Reduced Representation Sequencing approach allowed exploration of the genetic variability of both genotypes to identify candidate genes that could regulate stress responses. • Both abiotic stresses had a severe impact on plant growth parameters and on the reproductive phase of development. Growth parameters and leaf gas exchange measurements revealed that the two genotypes used different physiological strategies to overcome individual and combined stresses, with E42 having a more efficient capacity to utilize the limiting water resources. Activation of antioxidant defence mechanisms seemed to be critical for both genotypes to counteract combined abiotic stresses. Candidate genes were identified that could explain the different physiological responses to stress observed in E42 compared with LA3120. • Results here obtained have shown how new tomato genetic resources can be a valuable source of traits for adaptation to combined abiotic stresses and should be used in breeding programmes to improve stress tolerance in commercial varieties.
... For most tomato cultivars, if the temperatures exceed 26ºC during the day and 20ºC during the night, fruit set is impaired mainly due to floral development alteration, and therefore fruit yield is reduced (Sherzod et al., 2020). Because the response to stress varies widely between genotypes (Arena et al., 2020;Gonzalo et al., 2021), gaining deeper knowledge of the developmental and molecular response of various cultivars may provide clues for innovative breeding strategies aiming at improving commercial tomato cultivars for better adaptability to climate change. In the present work, we investigated the responses to heat stress in eleven tomato cultivars producing fruits of different sizes and shapes. ...
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Climate change is one of the biggest threats that human society currently needs to face. Heat waves associated with global warming negatively affect plant growth and development and will increase in frequency. Tomato is one of the most produced and consumed fruit in the world but remarkable yield losses occur every year due to the sensitivity of many cultivars to heat stress. New insights into how tomato plants are responding to heat waves will contribute to the development of new cultivars with high yields under harsh temperature conditions. In this study, the analysis of microsporogenesis and pollen germination rate of eleven tomato cultivars after exposure to a simulated heat wave revealed differences between genotypes. The transcriptome of floral buds at two developmental stages of five cultivars selected based on their pollen germination tolerance or sensitivity, revealed common and specific molecular responses implemented by tomato cultivars to cope with heat waves. These data provide valuable insights into the underlying molecular adaptation of floral buds to heat stress and will contribute to the development of future climate resilient tomato varieties.
... The dry biomass was determined by drying the above ground biomass (leaves and stems) in a forced-air oven at 75 • C until constant weight. SLW was measured in 5 leaves per treatment as the ratio between leaf dry mass and leaf area [25] on six fully expanded leaves. ...
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Salinity in water and soil is among the major constraints to the cultivation of ornamental crops since it can affect their growth and aesthetic value. A greenhouse experiment was carried out to assess whether the application of two different salts (80mMNaCl or 53.3mMCaCl2, with a final ionic concentration of 160 mM) could differently modulate the anatomical and physiological acclimation of an important ornamental species such as Viburnum tinus L. var. lucidum. Eco-physiological analyses (e.g., leaf gas exchange and chlorophyll a fluorescence emission) were performed and leaves were subjected to light microscopy analysis to quantify functional anatomical traits through digital image analysis. Results showed that the two iso-osmotic solutions induced different structuremediated physiological alterations in V. tinus plants. Photosynthesis was lowered by CaCl2 treatments (􀀀58%) more than by NaCl (􀀀37%), also due to the occurrence of photodamage apart from stomatal limitations. Neither Na+ nor Cl􀀀 exhibited toxic effects in leaf lamina structure which was reflected in the limited reduction in dry matter accumulation. Overall data were interpreted focusing on the coordination among leaf structural and functional traits suggesting that the fine control of functional anatomical traits contributes to physiological acclimation to both stressful conditions.
... For the chlorophyll a fluorescence measurements, a portable FluorPen FP100max fluorometer, equipped with a light sensor (Photon System Instruments, Brno, Czech Republic) was used following the procedure reported in Arena et al. (2020) [31]. The ground fluorescence (F 0 ) was induced by an internal LED blue light (1-2 µmol photons m −2 s −1 ) on 30 min dark-adapted leaves. ...
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The aim of this work was to assess the possible coordination mechanisms between leaf functional anatomical traits and physiological acclimation of potted Callistemon citrinus plants when subjected to two saline iso-osmotic solutions using NaCl or CaCl2 . Digital image analysis was applied to quantify anatomical parameters with a specific focus on the occurrence of signs of structural damage as well as on alterations, such as the occurrence of shrunk tissues and accumulation of phenolic compounds. Morpho-anatomical analyses were accompanied by measurements of leaf gas exchange and chlorophyll fluorescence emission. Results showed that C. citrinus plants, when irrigated with the CaCl2 solution, induced a leaf morpho-anatomical structure which did not allow the maintenance of high photosynthetic performance under such conditions, compared to both controls and plants treated with the iso-osmotic solution of NaCl. Data collected seem to suggest a close relationship between anatomical traits and eco-physiological strategies in maintaining the photosynthetic efficiency under saline stress conditions and highlight the fundamental role of leaf anatomy in imposing the limits of plant physiology.
... We assessed the post-harvest properties of these 41 accessions, such as the duration of fruit shelf life and fungal susceptibility, under normal and high temperature regimes. These genotypes have been extensively characterised for their responses to elevated temperature and/or drought growth conditions using Qualitative Trait Loci analysis, high-throughput geno-and phenotyping, assessment of flowering, fruit set, yield, fruit nutritional composition, antioxidant and antiinflammatory activities, as well as by screening for photosynthetic efficiency [23][24][25][26][27][28][29][30][31][32][33][34]. In addition, several candidate genes regulating heat stress tolerance were recently identified in varieties from this collection [24,29]. ...
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The fleshy fruit of tomato (Solanum lycopersicum) is a commodity used worldwide as a fresh or processed product. Like many crops, tomato plants and harvested fruits are susceptible to the onset of climate change. Temperature plays a key role in tomato fruit production and ripening, including softening, development of fruit colour, flavour and aroma. The combination of climate change and the drive to reduce carbon emission and energy consumption is likely to affect tomato post-harvest storage conditions. In this study, we investigated the effect of an elevated storage temperature on tomato shelf life and fungal susceptibility. A collection of 41 genotypes with low and high field performance at elevated temperature, including different growth, fruit and market types, was used to assess post-harvest performances. A temperature increase from 18–20 °C to 26 °C reduced average shelf life of fruit by 4 days ± 1 day and increased fungal susceptibility by 11% ± 5% across all genotypes. We identified tomato varieties that exhibit both favourable post-harvest fruit quality and high field performance at elevated temperature. This work contributes to efforts to enhance crop resilience by selecting for thermotolerance combined with traits suitable to maintain and improve fruit quality, shelf life and pathogen susceptibility under changing climate conditions.
... With the increase of temperature, plants enhance their transpiration rate due to the water vapor gradient and need for cooling, this results in transpiration losses, stomata closure, and photosynthesis decrease [9,35]. In cultivated tomato, heat is responsible for a decrease in photosynthesis, flower wilting, low pollen viability, low fruit set, and a decrease in overall crop yield [40,41]. Heat is frequently associated with drought, which demonstrates the value of studying the combination of both stresses [10]. ...
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Within the tomato clade, Solanum chilense is considered one of the most promising sources of genes for tomato (S. lycopersicum) selection to biotic and abiotic stresses. In this study, we compared the effects of drought, high temperature, and their combination in two cultivars of S. lycopersicum and six populations of S. chilense, differing in their local habitat. Plants were grown at 21/19 °C or 28/26 °C under well-watered and water-stressed conditions. Plant growth, physiological responses, and expression of stress-responsive genes were investigated. Our results demonstrated strong variability among accessions. Differences in plant growth parameters were even higher among S. chilense populations than between species. The effects of water stress, high temperature, and their combination also differed according to the accession, suggesting differences in stress resistance between species and populations. Overall, water stress affected plants more negatively than temperature from a morpho-physiological point of view, while the expression of stress-responsive genes was more affected by temperature than by water stress. Accessions clustered in two groups regarding resistance to water stress and high temperature. The sensitive group included the S. lycopersicum cultivars and the S. chilense populations LA2931 and LA1930, and the resistant group included the S. chilense populations LA1958, LA2880, LA2765, and LA4107. Our results suggested that resistance traits were not particularly related to the environmental conditions in the natural habitat of the populations. The expression of stress-responsive genes was more stable in resistant accessions than in sensitive ones in response to water stress and high temperature. Altogether, our results suggest that water stress and high temperature resistance in S. chilense did not depend on single traits but on a combination of morphological, physiological, and genetic traits.
... Such germplasm could be useful to study multiple stress resilience mechanism(s) to develop solanaceous varieties for organic or low-input agriculture. Combined multiple stress tolerance is of great interest [40,41] and there are efforts to achieve resilient tomatoes that keep unique flavor [42,43]. Therefore, valorization of collected accessions with a focus on stress tolerance and description of locally adapted flavor would be of great interest. ...
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The present work compiles the results of three-year expeditions organized between 2011 and 2013 aimed at plant crop collection in the area of Lithuania, an Eastern European country. Accessions of fruit trees, vegetables, cereals, forage, industrial, fibrous, medical, and spice crops as well as accompanying segetal plants were collected in 5 ethnographic regions of Lithuania. In total, 1010 samples of seeds, bulbs, and plant grafts were obtained. The majority of the collected samples belonged to the Cucurbitaceae, Fabaceae, Solanaceae, Alliaceae, and Rosaceae families. The accessions were described and deposited in the long-term storage in Lithuanian and Polish Gene Banks. Almost all collected cucumbers, pumpkins, squashes, and oat plants were morphologically characterized in field experiments. Cucurbita pepo accessions showed high morphological diversity, while limited diversity of cucumber accessions was observed. Oat plants were characterized by high morphological diversity and resistance to diseases present in some of the investigated accessions. Further characterization of collected apple trees and other accessions is underway. Obtaining germplasm in the area of Lithuania fulfills a gap in current plant crop collections. Collected material could be valuable for pre-breeding evaluation and further breeding programs as well as the study of genetic diversity.
... The fluorescence parameters and images were determined at 7:00-11:00 a.m. after 0, 4, 8, 12, 16 and 20 days of submergence. The chlorophyll fluorescence measurements were taken using a Fluorcam portable chlorophyll fluorescence imager (Handy Fluorcam FC 1000-H, PSI, Brno, Czech Republic) from the top 3-5 fully expanded leaves of the mulberry seedlings, which revealed the PS II activity 39,40 . The parameters are set to shutter, 1; sensitivity, 28%; super, 90%; far, 50%; act, 1 100%; and QY_max, a range of 0-0.67. ...
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Because of its high flooding tolerance, in recent years, mulberry has become a tree species that is used in plant restoration in impact zones in reservoir areas. Therefore, 3-year-old potted forage mulberry seedlings were used to investigate the mechanism of mulberry adaptation to flooding stress. An indoor simulated flooding method was adopted to study the morphology of mulberry seedling leaves and the changes in leaf chlorophyll fluorescence parameters and fluorescence imaging under different flooding times and depths. The results showed that the leaves of mulberry seedlings treated with shallow submergence remained healthy during the flooding period, while the leaves of mulberry seedlings treated with half submergence and full submergence showed different degrees of waterlogging symptoms in the middle and late flooding periods and formed adventitious roots at the base of the stem. Most of the chlorophyll fluorescence parameters decreased at the beginning of flooding, but the steady-state degree of closure of PS II reaction centres (1-qP_Lss) increased significantly. In the later stage of flooding, the fluorescence parameters showed relatively stable trends. Based on these results, we conclude that mulberry has high flooding tolerance due to a combination of morphological and physiological responses.
Tomato, an important vegetable crop cultivated worldwide is an essential component of the Ghanaian diet. However, production in Ghana is hampered by heat stress (HS) during certain periods of the year. Tomato is sensitive to HS, especially during the reproductive stage, inducing pollen sterility, floral and fruit abortion, and culminating in low yield. This necessitates development of HS-tolerant tomato varieties. The HS-tolerance potential of 13 tomato lines was assessed under both poly-house and open field conditions at day temperatures of ~35°C. Evaluation was based on agro-morphological and physiological traits. Most lines had their highest fruit yield (FY) under poly-house conditions, but declined drastically (up to 92%) in the field when day/night temperatures reached 40/28°C respectively. A second field experiment revealed that the low FY was due to poor fruit set despite the many flowers produced. Correlations between days to 50% fruiting and physiological traits (ΦII, chlorophyll content, and leaf temperature) were high and significant, whereas those between FY and all physiological parameters were negligible. Lines KAC13 and KAU03 consistently had high FY, ΦII, and chlorophyll content. Nevertheless, KAU01 had the highest FY in two out of three experiments in addition to having the highest percentage fruit set. The lines that show high potential tolerance to HS may be incorporated into breeding programmes to further improve the tolerance of tomatoes to HS.
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Abiotic stresses can cause a substantial decline in fruit quality due to negative impacts on plant growth, physiology and reproduction. The objective of this study was to verify if the use of a biostimulant based on plant and yeast extracts, rich in amino acids and that contains microelements (boron, zinc and manganese) can ensure good crop yield and quality in tomato plants grown at elevated temperatures (up to 42 °C). We investigated physiological responses of four different tomato landraces that were cultivated under plastic tunnel and treated with the biostimulant CycoFlow. The application of the biostimulant stimulated growth (plants up to 48.5% taller) and number of fruits (up to 105.3%). In plants treated with the biostimulant, antioxidants contents were higher compared to non-treated plants, both in leaves and in fruits. In particular, the content of ascorbic acid increased after treatments with CycoFlow. For almost all the traits studied, the effect of the biostimulant depended on the genotype it was applied on. Altogether, the use of the biostimulant on tomato plants led to better plant performances at elevated temperatures, that could be attributed also to a stronger antioxidant defence system, and to a better fruit nutritional quality.
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High temperature is one of the most detrimental abiotic stresses in tomatoes. Many studies highlighted that even small increases in temperature can alter the plant reproductive system, causing a significant reduction in tomato yield. The aim of this study was to exploit the phenotypic and genomic variations of a tomato landrace collection grown at high temperatures. Fifteen genotypes were selected as the best performing in two experimental fields. The selection was based on six yield-related traits, including flower earliness, number of flowers per inflorescence, fruit set, number of fruit per plant, fruit weight and yield per plant. In order to identify markers targeting traits that could be highly influenced by adverse climate conditions, such as flowering and fruit setting, an association mapping approach was undertaken exploiting a tomato high-throughput genomic array. The phenotypic variability observed allowed us to identify a total of 15 common markers associated with the studied traits. In particular, the most relevant associations co-localized with genes involved in the floral structure development, such as the style2.1 gene, or with genes directly involved in the response to abiotic stresses. These promising candidate genes will be functionally validated and transferred to a cultivated tomato to improve its performance under high temperatures
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Drought (water deficits) and heat (high temperatures) stress are the prime abiotic constraints, under the current and climate change scenario in future. Any further increase in the occurrence, and extremity of these stresses, either individually or in combination, would severely reduce the crop productivity and food security, globally. Although, they obstruct productivity at all crop growth stages, the extent of damage at reproductive phase of crop growth, mainly the seed filling phase, is critical and causes considerable yield losses. Drought and heat stress substantially affect the seed yields by reducing seed size and number, eventually affecting the commercial trait ‘100 seed weight’ and seed quality. Seed filling is influenced by various metabolic processes occurring in the leaves, especially production and translocation of photoassimilates, importing precursors for biosynthesis of seed reserves, minerals and other functional constituents. These processes are highly sensitive to drought and heat, due to involvement of array of diverse enzymes and transporters, located in the leaves and seeds. We highlight here the findings in various food crops showing how their seed composition is drastically impacted at various cellular levels due to drought and heat stresses, applied separately, or in combination. The combined stresses are extremely detrimental for seed yield and its quality, and thus need more attention. Understanding the precise target sites regulating seed filling events in leaves and seeds, and how they are affected by abiotic stresses, is imperative to enhance the seed quality. It is vital to know the physiological, biochemical and genetic mechanisms, which govern the various seed filling events under stress environments, to devise strategies to improve stress tolerance. Converging modern advances in physiology, biochemistry and biotechnology, especially the “omics” technologies might provide a strong impetus to research on this aspect. Such application, along with effective agronomic management system would pave the way in developing crop genotypes/varieties with improved productivity under drought and/or heat stresses.
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Elevated atmospheric CO2 improves leaf photosynthesis and plant tolerance to heat stress, however, the underlying mechanisms remain unclear. In this study, we exposed tomato plants to elevated CO2 (800 μmol mol-1) and/or high temperature (42∘C for 24 h), and examined a range of photosynthetic and chlorophyll fluorescence parameters as well as cellular redox state to better understand the response of photosystem II (PSII) and PSI to elevated CO2 and heat stress. The results showed that, while the heat stress drastically decreased the net photosynthetic rate (Pn), maximum carboxylation rate (Vcmax), maximum ribulose-1,5-bis-phosphate (RuBP) regeneration rate (Jmax) and maximal photochemical efficiency of PSII (Fv/Fm), the elevated CO2 improved those parameters under heat stress and at a 24 h recovery. Furthermore, the heat stress decreased the absorption flux, trapped energy flux, electron transport, energy dissipation per PSII cross section, while the elevated CO2 had the opposing effects that eventually decreased photoinhibition, damage to photosystems and reactive oxygen species accumulation. Similarly, the elevated CO2 helped the plants to maintain a reduced redox state as evidenced by the increased ratios of ASA:DHA and GSH:GSSG under heat stress and at recovery. Furthermore, the concentration of NADP+ and ratio of NADP+ to NADPH were induced by elevated CO2 at recovery. This study unraveled the crucial mechanisms of elevated CO2-mediated changes in energy fluxes, electron transport and redox homeostasis under heat stress, and shed new light on the responses of tomato plants to combined heat and elevated CO2.
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This study aimed to phenotype young tomato (Solanum lycopersicum L.) plants for heat tolerance by measuring Fv/Fm after short-term heat treatments in climate chambers and selected sensitive (low Fv/Fm) and tolerant (high Fv/Fm) cultivars to investigate their in-field performance. Twenty-eight genotypes were phenotyped at 40: 28°C for 2 days in climate chambers. A second screening (four high Fv/Fm and four low Fv/Fm genotypes) was conducted for 4 days at 38: 28°C, followed by 5 days' recovery (26: 20°C). The tolerant genotypes maintained high net photosynthesis (PN) and increased stomatal conductance (gs) at 38°C, allowing better leaf cooling. Sensitive genotypes had lower Fv/Fm and PN at 38°C, and gs increased less than in the tolerant group, reducing leaf cooling. Under controlled conditions, all eight genotypes had the same plant size and pollen viability, but after heat stress, plant size and pollen viability reduced dramatically in the sensitive group. Two tolerant and two sensitive genotypes were grown in the field during a heat wave (38: 26°C). Tolerant genotypes accumulated more biomass, had a lower heat injury index and higher fruit yield. To our knowledge, this is the first time screening for heat tolerance by Fv/Fm in climate chambers was verified by a field trial under natural heat stress. The differences after heat stress in controlled environments were comparable to those in yield between tolerant and sensitive groups under heat stress in the field. The results suggest that Fv/Fm is effective for early detection of heat tolerance, and screening seedlings for heat sensitivity can speed crop improvement.
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Despite a range of initiatives to reduce global carbon emission, the mean global temperature is increasing due to climate change. Since rising temperatures pose a serious threat of food insecurity, it is important to further explore important biological molecules that can confer thermotolerance to plants. Recently, melatonin has emerged as a universal abiotic stress regulator that can enhance plant tolerance to high temperature. Nonetheless, such regulatory roles of melatonin were unraveled mainly by assessing the effect of exogenous melatonin on plant tolerance to abiotic stress. Here, we generated melatonin deficient tomato plants by silencing of a melatonin biosynthetic gene, CAFFEIC ACID O-METHYLTRANSFERASE 1 (COMT1), to unveil the role of endogenous melatonin in photosynthesis under heat stress. We examined photosynthetic pigment content, leaf gas exchange, and a range of chlorophyll fluorescence parameters. The results showed that silencing of COMT1 aggravated heat stress by inhibiting both the light reactions and the carbon fixation reactions of photosynthesis. The photosynthetic pigment content, light absorption flux, trapped energy flux, energy dissipation, density of active reaction center per photosystem II (PSII) cross-section, the photosynthetic electron transport rate, the maximum photochemical efficiency of PSII photochemistry, and the rate of CO2 assimilation all decreased in COMT1-silenced plants compared with that of non-silenced plants particularly under heat stress. However, exogenous melatonin alleviated heat-induced photosynthetic inhibition in both genotypes, indicating that melatonin is essential for maintaining photosynthetic capacity under stressful conditions. These findings provide genetic evidence on the vital role of melatonin in photosynthesis and thus may have useful implication in horticultural crop management in the face of climate change.
Mediterranean tomato landraces adapted to arid environments represent an option to counteract drought, and to address the complexity of responses to water deficit and recovery, which is a crucial component of plant adaptation mechanisms. We investigated physiological, biochemical and molecular responses of two Mediterranean tomato landraces, “Locale di Salina” (Lc) and “Pizzutello di Sciacca” (Pz) under two dehydration periods and intermediate rehydration in greenhouse pot‐experiments. Relationship between CO2 assimilation and stomatal conductance under severe water stress (gs < 0.05 mol m⁻² s⁻¹) indicated the occurrence of stomatal and non‐stomatal limitations of photosynthesis. Gas‐exchanges promptly recovered within 2‐3‐day re‐hydration. ABA and gs showed a strict exponential relationship. Both leaf ABA and proline peaked under severe water stress. Lc showed higher accumulation of ABA and higher induction of the expression of both NCED and P5CS genes than Pz. PARP increased during imposition of stress, mainly in Lc, and decreased under severe water stress. The two landraces hardly differed in their physiological performance. Under severe water stress, stomatal conductance showed low sensitivity to ABA, which instead controlled stomatal closure under moderate water stress (gs > 0.15 mol m⁻² s⁻¹). The prompt recovery after re‐dehydration of both landraces confirmed their drought tolerant behaviour. Differences between the two landraces were instead observed at biochemical and molecular levels. This article is protected by copyright. All rights reserved.
Tomato cultivation at lower or higher temperatures than the optimum negatively affects plant growth and development. Large differences in abiotic stress tolerance have been found between Solanum lycopersicum and wild tomato species. Our aim was to compare temperature stress tolerance in cultivated and wild tomato genotypes to identify cold- and heat-tolerant tomatoes for further utilization in tomato breeding. The maintained net photosynthetic rate (PN) and chlorophyll fluorescence was related to the tolerance of tomatoes at temperature stress. The PN and chlorophyll fluorescence of one cultivated tomato (Ly from S. lycopersicum) and six wild tomatoes genotypes (Ha from Solanum habrochaites, Pe from Solanum pennellii, Pi1 and Pi2 from Solanum pimpinellifolium, Pr1 and Pr2 from Solanum peruvianum) grown at low (12 °C) and high (33 °C) temperatures were compared. The PN of four tomato genotypes during temperature stress were lower than the control, but not in Pe, Pr1, and Pr2. The maximum quantum efficiency of photosystem II (Fv/Fm) of the cultivated tomatoes was lower at both 12 and 33 °C than the control using Handy PEA, whereas Fv/Fm using MINI-PAM was lower only at 12 °C. The chlorophyll fluorescence OJIP transient (OJIP curve) revealed differences between temperature stress responses and tomato genotype. With the exception of Pr2, the Fv/Fm in wild tomatoes was unaffected by temperature stress; however, they still maintained clear genotype differences for other physiological traits such as PN, quantum yield of PSII (Fq′/Fm′), electron transport rate, non-photochemical quenching, and the fraction of open PSII centers (qL). These results indicated that the wild tomato varieties Pe and Pr1 had the highest temperature stress tolerance, while the cultivated species was the more sensitive to temperature stress in comparison. In general, the wild tomato genotypes were more tolerant to both cold and heat stress than the cultivated tomato, suggesting that these wild species could be used to uncover underlying mechanisms of temperature stress tolerance and will be promising sources of genetic variability for temperature stress tolerance in breeding programs.