Journal of Plant Physiology 246–247 (2020) 153130

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Journal of Plant Physiology

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

Photochemistry and photoprotection of ‘Gem’ avocado (Persea americana T

Mill.) leaves within and outside the canopy and the relationship with fruit
maturity
Sabelo Shezia, Lembe Samukelo Magwazaa,b,*, Jacob Mashiloa,b, Samson Zeray Tesfaya,
Asanda Mditshwaa
a
Discipline of Horticultural Science, School of Agricultural, Earth and Environmental Sciences, University of KwaZulu-Natal, Private Bag X01, Scottsville, 3209,
Pietermaritzburg, South Africa
b
Discipline of Crop Science, School of Agricultural, Earth and Environmental Sciences, University of KwaZulu-Natal, Private Bag X01, Scottsville, 3209, Pietermaritzburg,
South Africa

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

Keywords: A reduction in photosynthesis results in a reduced CO2 assimilation rate and availability of carbohydrates es-
Photosynthetic efficiency sential for fruit growth and development. This study determined photosynthetic efficiency and photoprotection
Photoprotection mechanisms within and outside leaf canopy positions in ‘Gem’ avocado orchards and their relationship with
Avocado maturity avocado fruit maturity. The study was conducted in a commercial orchard at Everdon Estate in KwaZulu-Natal,
Dry matter content
South Africa. A total of 15 eight-year-old avocado trees (cv. Gem) were selected in a completely randomised
Canopy positions
design with three replicates, with each replicate consisting of five trees. Data were collected bi-weekly on
photosynthetic rate (A), effective quantum efficiency of photosystem II (ϕPSII), stomatal conductance (gs),
transpiration rate (T), electron transport rate (ETR), minimum fluorescence (Fo’), maximum fluorescence (Fm’),
variable fluorescence (Fv’), intrinsic water use efficiency (WUEi), instantaneous water use efficiency (WUEins),
intercellular CO2 concentration (Ci) and photochemical quenching (qP) from full bloom to fruit physiological
maturity (∼25 % dry matter content (DM)). The results showed that leaves from the outside position had higher
A (29.46 mol CO2 m−2s-1); gs (0.078 mol CO2 m−2s-1); ΦPS II (0.32); and qP (0.52) compared to those within the
canopy position with lower A (19.27 mol CO2 m−2s-1); gs (0.0037 mol CO2 m−2s-1); ΦPS II (0.044) and qP
(0.075), respectively. Contrastingly, chlorophyll fluorescence and photoprotection parameters were higher
within the canopy than on the outside, suggesting that the greater proportion of energy accumulated within the
canopy was used for photoprotection other than photochemistry. Photosynthetic rate (A), gs, Ci, T, WUEi and
WUEins, correlated significantly with mesocarp dry matter (DM), while all other parameters correlated poorly.
The high photosynthetic efficiency of leaves from outside the canopy resulted in an average DM of 28.9 %
compared to 26.9 % of fruit within the canopy. The present findings suggest that reduced photosynthetic effi-
ciency of ‘Gem’ avocado within the canopy position does not compromise fruit DM by reserving more energy for
photoprotection; however, it delays maturity by about two weeks.

Abbreviations: gs, stomatal conductance; T, transpiration rate; A, photosynthetic efficiency; Fo’, minimum fluorescence; Fm’, maximum fluorescence; Fv’, variable
fluorescence; Fv’/Fm’, maximum quantum yield of PS II photochemistry; ϕPS II, effective quantum efficiency of photosystem II; qP, photochemical quenching and qN,
non-photochemical quenching; ETR, electron transport rate; 1-qp, proportion of open reaction canters; D, thermal heat dissipation; RHD, rate of thermal heat
dissipation; P, fraction of photon energy absorbed by PS II antennae that was trapped by “open” PSII reaction centres; RPC, rate of photochemistry; Y (NO),
photoinhibition of photosystem II reaction centres; Ci, intercellular CO2 concentration; A/Ci, ratio of photosynthetic efficiency and intercellular CO2 concentration;
WUEi, intrinsic water use efficiency; DM, dry matter content; Lx cycle, lutein epoxide cycle; Lx, lutein-5,6-epoxide; L, lutein; α-C, α-carotene; ROS, reactive oxygen
species; WUEins, instantaneous water use efficiency
⁎
Corresponding author at: Discipline of Horticultural Science, School of Agricultural, Earth and Environmental Sciences, University of KwaZulu-Natal, Private Bag
X01, Scottsville, 3209, Pietermaritzburg, South Africa.
E-mail address: Magwazal@ukzn.ac.za (L.S. Magwaza).

https://doi.org/10.1016/j.jplph.2020.153130
Received 9 October 2019; Received in revised form 1 February 2020; Accepted 2 February 2020
Available online 07 February 2020
0176-1617/ © 2020 Elsevier GmbH. All rights reserved.
S. Shezi, et al.
1. Introduction
Avocados (Persea americana Mill.) originated from Mexico and Latin
America, where orchards are exposed to slightly fluctuating daily and
annual temperatures. Mexican climatic conditions are also char-
acterised by constant light intensity and relative humidity throughout
the year. In South Africa the crop is cultivated under varied environ-
mental conditions that are totally different from its region of origin
(Bertling and Cowan, 1998; Powles, 1984). South Africa is categorised
as one of the driest countries in the world and production of various
crops occur under extreme dry and hot conditions. Variation in en-
vironmental conditions such as light intensity, temperature and relative
humidity under these conditions is likely to affect photosynthetic effi-
ciency, growth and maturity of avocado fruit. More specifically, a re-
duction in photosynthetic efficiency as a result of high fluxes of pho-
tosynthetically active radiation/light energy cause photoinhibition/
damage (Zeng et al., 2017).
Photoinhibition of photosystem II (PS II) occurs at the oxygen-
evolving complex (OEC) by the release of manganese (Mn) ions (Mn2+)
(Hakala et al., 2005; Zsiros et al., 2006). The disruption of the Mn
cluster upon light absorption results in thermal energy dissipation,
cyclic flow of electrons around PS I and photorespiration (Takahashi
and Badger, 2011), which then cause a reduced CO2 assimilation rate.
Photoinhibition of PS II reaction centres is prevented by avoiding light
absorption using the Mn cluster of the OEC and dissipating excess en-
ergy absorbed by photosynthetic pigments (Takahashi and Badger,
2011). Regardless of how long photoinhibition has occurred, any re-
duction in photosynthesis results in reduced assimilation rates and
hence availability of carbohydrates essential for growth and develop-
ment.
Primary changes in tree physiology can be assessed through mea-
surement of leaf chlorophyll fluorescence and gas exchange parameters
(Flexas et al., 1999; Mashilo et al., 2017). The measurement of chlor-
ophyll fluorescence is useful in understanding photoprotection me-
chanisms associated with the removal of excess excitation energy, ei-
ther as heat or fluorescence, from the photosynthetic apparatus (Araus
et al., 1998). Changes in photosynthetic efficiency (A), stomatal con-
ductance (gs), transpiration rate (T), maximum quantum yield of PS II
photochemistry (Fv’/Fm’), the effective quantum efficiency of photo-
system II (ϕPS II), photochemical quenching (qP) and non-photo-
chemical quenching (qN), electron transport rate (ETR) and proportion
of open reaction canters (1-qp), are some key physiological indicators of
photosynthetic efficiency and abiotic stress tolerance (i.e. light, heat
and drought) (Mashilo et al., 2017). Furthermore, physiological mea-
sures such as photoinhibition of PS II reaction centres (Y), the fraction
of photon energy trapped by “open” PS II reaction centres (1-qP) and
utilized in PS II photochemistry (P), the portion of the absorbed photon
energy that was thermally dissipated (D), rate of photochemistry (RPC)
and rate of heat dissipation (RHD) are key parameters for assessing
photoinhibition of photosynthetic apparatus.
Generally, avocado leaves that are fully exposed to the sun receive
more irradiation than the ones that are partially or fully shaded (Woolf
et al., 1999). Therefore, leaf canopy position significantly affects the
overall photosynthetic efficiency, with leaves within the canopy
showing lower photosynthetic rates than leaves outside the canopy
(Oguchi et al., 2018; Shezi et al., 2019). These findings suggest that
photodamage is more likely to occur in the leaves that are fully exposed
to the sun (i.e. exposure to high light intensity results in production of
reactive oxygen species, known as ROS, which disturb chlorophyll
functioning) than in leaves that are shaded (Ruban et al., 2012). As a
result, photosynthetic capacity and photoprotection mechanisms within
and outside canopy positions may vary with differences in the light
intensity, which may affect the rate of avocado fruit growth, develop-
ment and maturity. These may be affected because the energy that is
used for photoprotection is partitioned from the total amount of light
energy absorbed/intercepted by a leaf to be used for photochemistry
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Journal of Plant Physiology 246–247 (2020) 153130
(Maxwell and Johnson, 2000). Energy partitioning among photo-
chemistry and photoprotection occurs unevenly and thus becomes one
of the sources of variation in the amount of assimilates accumulated
and translocated.
Furthermore, the higher rate of heat dissipation (RHD) in the leaves
from the outside canopy could be due to the de-epoxidation state of the
xanthophyll cycle pigments, which is known to be higher in leaves from
the outside canopy because of high light intensity. Filella et al. (2009)
reported that the de-epoxidation state (DEPS) and violaxanthin, an-
theraxanthin and zeaxanthin (V + A+Z)/chlorophyll (a+b) of Scots
pine (Pinus sylvestris L.) and Holm oak (Quercus ilex L.) were higher in
the leaves that were fully exposed to the sun than the shade leaves. The
higher de-epoxidation of the xanthophyll cycle in the leaves from the
outside canopy was also linked to the higher β-carotene and total car-
otenoid contents (Filella et al., 2009). Occurrence of photoprotection
mechanisms such as the xanthophyll cycle consume energy and thus
could alter the distribution of assimilates towards fruit that are exposed
or the ones fully shaded.
Uneven distribution of assimilates results in varying growth and
development and hence maturity of avocado fruits. Furthermore, a
strong sink strength of fruit that are advanced in terms of development,
drives the uneven distribution of assimilates, because they outcompete
the ones with a lesser sink strength (Fischer et al., 2012). This results in
an uneven maturity. Unfortunately, avocado fruit does not show any
signs of maturity (Magwaza and Tesfay, 2015; Olarewaju et al., 2016;
Ncama et al., 2018), and uneven maturity can only be detected later
after fruit has been harvested and kept for ripening. Maturity of avo-
cado fruit (cv. Carmen) harvested from the outside canopy occurs ear-
lier than in fruit that were harvested within the canopy, resulting in one
of the causes of uneven ripening (Shezi et al., 2019). This is because the
chances are that fruit from the same tree are harvested on the same day
and probably dispatched in the same pallet for export. During post-
harvest storage and operations such as ripening fruits using ethylene,
fruits ripen unevenly because their biochemical compounds that code
for maturity vary according to their canopy positions. In avocado fruit,
mesocarp oil and dry matter content (DM) are the major biochemical
markers that have been accepted globally as the best parameters that
code for optimum maturity (Woolf et al., 2003; Ncama et al., 2018).
Therefore, the DM is used commercially, because it is strong and po-
sitively correlates with oil content and is determined easily.
Previous studies have revealed that avocado maturity is linked to
the intercepted light energy (Whiley et al., 1992; Shezi et al., 2019).
However, this energy is partitioned into either of the three phases, i.e.
used for photochemistry, or for photoprotection (where it is emitted as
heat), or it is re-emitted as fluorescence (Maxwell and Johnson, 2000).
Therefore, knowledge of the key processes such as photosynthetic ac-
tivity and photoprotection mechanisms of commercial avocado culti-
vars such as ‘Gem’, which often experience excess light, can be useful
when discussing the causes of uneven growth, development and ma-
turity which result in uneven ripening. Furthermore, understanding the
proportions of partitioned light energy can aid in deducing if the oc-
currence of uneven ripening in ‘Gem’ avocado fruit can be controlled or
not. Therefore, this study aimed to establish to what extent ‘Gem’
avocado trees can compensate energy for photoprotection within and
outside the canopy, and how that affects maturity of avocado fruit. The
objective was to determine the photosynthetic efficiency and photo-
protection within and outside the leaf canopy positions in ‘Gem’ avo-
cado trees and whether these have a relationship with fruit maturity.
2. Materials and methods
2.1. Plant material
The study was conducted in a commercial orchard at Westfalia Fruit
Everdon Estate in the KwaZulu-Natal midlands (29°27’14.4’’S’
30°16’40.8’’E), South Africa, during the 2018/19 season. A total of 15
S. Shezi, et al.
eight-year-old avocado trees (cv. Gem) were selected in a completely
randomised design with three replicates, each consisting of five trees.
The study was conducted during the autumn, winter and spring seasons
(February 2018 to January 2019). Data were collected bi-weekly from
full bloom to fruit physiological maturity (25 % DM).
2.2. Orchard management
The experiment was conducted using eight-year-old ‘Gem’ avocado
cultivars grafted onto Dusa® rootstock. Trees were spaced 4 m within
rows and 7 m between rows on a slight slope of about 5 %. The orchard
was irrigated using an automated micro-sprinkler to maintain soil at
80–100 % field capacity, based on AquaHous Irrigation Probe re-
quirements. Therefore, there were no specific water volumes and tim-
ings, as irrigation started at any time after a message reception that the
water level was below 80 % field capacity. Since the orchard was only
eight years old, no training system, shoot thinning and fruit thinning
techniques were performed during the season. Copper (6 g/L using an
electrostatics injector) was applied twice during the year, in January
and November, to control blackspot. Scouting was done at a frequency
of three days a week (Mondays, Thursdays and Fridays) for identifying
fruit flies, mainly Ceratitis capitata (Medfly), C. cosyra (Marula fly), C.
rosa (Natal fly) and Bactrocera invadens (Invader fly). In addition to
scouting, trapping was also done once a week, targeting fruit flies
(Ceratitis capitata) and false codling moth (Thaumatotibia leucotreta).
Temperature and rainfall were recorded continuously (Fig. 1).
2.3. Experiment structure and design
The experiment was conducted on two canopy positions: within and
outside the canopy. Within the canopy, the position was characterised
by fully shaded leaves, while outside the canopy the position was
characterised by leaves fully exposed to the sun. Discrimination of
leaves according to canopy position in this study was done based on the
orientation (upper or lower, and fully exposed to the sun or fully
shaded) of leaves on the tree. Leaves within the canopy position are
orientated at the lower edge of a tree, and are always fully shaded,
while leaves from the outside canopy are the upper leaves of the tree
that are always fully exposed to the sun. An experiment was set up in a
completely randomised design with three replicates of five trees each.
The data were collected only on fully mature photosynthesising leaves.
2.4. Measurement of leaf chlorophyll fluorescence and gas exchange
parameters
Leaf gas exchange and chlorophyll fluorescence were measured si-
multaneously using the LI-6400 XT Portable Photosynthesis System
(Licor Bioscience, Inc. Lincoln, Nebraska, USA) fitted with an infrared
gas analyser attached to a leaf chamber fluorometer (LCF) (6400-40B, 2
Fig. 1. The minimum temperature (°C), maximum temperature (°C) and total
rainfall (mm) recorded at the experimental site during February 2018 to
January 2019 (Shezi et al., 2019).
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Journal of Plant Physiology 246–247 (2020) 153130
cm2 leaf area, Licor Bioscience, Inc. Lincoln, Nebraska, USA). The re-
ference CO2 was fixed at 400, and the artificial saturating photo-
synthetic active radiation (PAR) was fixed at 1000 mmol m−2 s-1. Data
were collected from five leaves on non-stressed experimental trees.
Stomatal conductance (gs), transpiration rate (T), photosynthetic effi-
ciency (A), initial fluorescence (F0), maximum fluorescence (Fm),
maximum quantum yield of PS II photochemistry (Fv’/Fm)’, the effec-
tive quantum efficiency of photosystem II (ϕ PS II), photochemical
quenching (qP) and non-photochemical quenching (qN), electron
transport rate (ETR) and proportion of open reaction canters (1-qP)
were estimated within and outside the canopy (Demmig-Adams and
Adams Iii, 1992; Lichtenthaler et al., 2005; Klughammer and Schreiber,
2008; Mashilo et al., 2018).
Leaf gas exchange was measured in the outer tree canopy and
within the canopy. Parameters such as stomatal conductance (gs),
transpiration rate (T), and intercellular CO2 concentration (Ci) were
measured (Mashilo et al., 2018). In addition, the intrinsic water use
efficiency (WUEi), the instantaneous water use efficiency (WUEins), the
ratio of photosynthetic efficiency and intercellular CO2 concentration
(A/Ci) were calculated (Martin and Ruiz-Torres, 1992). The WUE was
calculated as per the Eqs. (1) and (2) below: intrinsic water use effi-
ciency (WUEi) and instantaneous water use efficiency (WUEins)
WUEi = A/gs (1)
WUEins = A/T (2)
Data collection took place between 11h00 and 13h00 on the clear
and sunny days. The data were measured once in each of the five leaves
within and outside the canopy of fifteen trees.
The measured chlorophyll fluorescence parameters were then used
to calculate some photochemical variables, which included Fv’/Fm’, the
maximum quantum efficiency of photosystem II photochemistry (ϕ PS
II), photochemical quenching (qP), non-photochemical quenching (qN)
and linear electron transport rate (LETR), calculated according to
Maxwell and Johnson (2000). The proportion of open photosystem II
(1-qP) reaction centres was calculated according to Demmig-Adams and
Adams Iii (1992).
Photoinhibition of photosystem II reaction centres (Y (NO)), defined
as the inability to protect itself against damage from excess light, was
calculated according to the Eq. (3) below.
Y (NO) = Fs/Fm’ (3)
The fraction of photon energy that was absorbed by photosystem II
antennae that was trapped by open reaction centres and used in PSII
photchemistry was calculated using Eq. (4) below.
P = Fv’/Fm’ – qP (4)
The fraction of absorbed photon energy that was thermally dis-
sipated was calculated as in Eq. (5) below.
D = 1-Fv’/Fm’ (5)
The rates of photochemistry and thermal heat dissipation were es-
timated using Eqs. (6) and (7), respectively.
RPD = P × PPDF (6)
RHD = D × PPDF (7)
2.5. Fruit sampling
Fruit were also sampled for DM, mesocarp dry matter content, as
one of the commercially used maturity indices. Sampling for DM started
from 182 days after full bloom to physiological maturity (25 % meso-
carp DM) or ripening without any form of shrivelling. The DM was
measured using a method described by Olarewaju et al. (2016). Sam-
ples of fresh mesocarp tissue were weighed, snap frozen in liquid
S. Shezi, et al.
Fig. 2. Photosynthetic rate (A), transpiration rate (T), stomatal conductance (gs) and intercellular CO2 concentration (Ci) of leaves in positions outside and within the
canopy of the ‘Gem’ avocado cultivar. Values are means ± SE (n = 30).
nitrogen and freeze dried to constant mass, using the Virtis Benchtop
freeze drier system (ES Model, SP Industries Inc., Warmister, USA), for
five days at 0.015 kPa and −75 °C. A total of nine fruit per canopy were
sampled on each sampling day. The DM was then calculated and ex-
pressed as a percentage.
2.6. Data analysis
Data on leaf chlorophyll fluorescence and gas exchange parameters
were subjected to the analysis of variance, using GenStat version 18th
Edition (VSN International, Hemel Hempstead, UK). Mean comparison
was done using Duncan’s multiple range test at 5 % level of sig-
nificance. Pearson’s correlation was used to determine the relationship
between the measured parameters.
3. Results
3.1. Chlorophyll fluorescence and gas exchange
Photosynthetic efficiency (A) differed significantly (p < 0.001) with
canopy positions (Fig. 2a). Leaves from the outside canopy had a higher
average A (29.5 mol CO2 m−2s-1) than those from inside the canopy,
which had a lower average A (19.3 mol CO2 m−2s-1). There was a
significant (p < 0.001) difference between transpiration rate (T) mea-
sured in different canopy positions (Fig. 2b). Average T was higher (8.3
mmol H2O m−2s-1) outside the canopy and lower (4.98 mmol H2O
m−2s-1) within the canopy. Stomatal conductance (gs) varied sig-
nificantly (p < 0.05) with the different canopy positions (Fig. 2c), with
the leaves from outside the canopy having a higher average gs (0.078
mol) CO2 m−2s-1 than the ones measured within the canopy (0.0037
mol CO2 m−2s-1). The average internal CO2 concentration (Ci) did not
differ (p > 0.05) with different canopy positions.
There was a highly significant difference (p < 0.001) in the intrinsic
water use efficiency (WUEi) that was measured in the different canopy
positions (Fig. 3). The WUEi was higher (349) within the canopy and
lower (180) outside canopy. The ratio of net assimilation and internal
CO2 concentration (A/Ci) had a significant variation (p < 0.001) among
different canopy positions. The A/Ci was measured higher outside the
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Journal of Plant Physiology 246–247 (2020) 153130
canopy (0.348) than within the canopy (0.235) (Fig. S1A). The in-
stantaneous water use efficiency (WUEins) also varied significantly
(p < 0.001) with different canopy positions (Fig. S1B). The WUEins was
higher within the canopy (7.780) and lower outside the canopy (4.26).
A highly significant (p < 0.001) variation was observed between the
LETR measured in different canopy positions (Fig. 3b). The LETR that
was measured outside the canopy was about seven times higher than
that measured inside the canopy, with averages of 158.0 and 22.0 for
outside and inside the canopy, respectively.
The minimum fluorescence (Fo’) and maximum fluorescence (Fm’)
differed significantly (p < 0.001) with the canopy positions (Fig. S2A
and S2B). The minimum fluorescence was higher inside the canopy
(597 μmol m−2 s-1) and lower outside the canopy (407 μmol m−2 s-1).
The maximum fluorescence had a higher average inside the canopy
(1524 μmol m−2 s-1) and a lower average outside the canopy (1089
μmol m−2 s-1). These results revealed high photosynthetic efficiency of
the leaves from the outside canopy in terms of photochemistry, com-
pared to the leaves from within the canopy.
The Fv’/Fm’ and Fv’ measured in different canopy positions differed
significantly (p < 0.001) (Fig. 4 and S2C). The Fv’ was higher within
the canopy, with an average of 926.6, and lower outside the canopy,
with an average of 681.6. The Fv’/Fm’ was recorded as higher in the
outside canopy, with an average of 0.612, and lower in the inside ca-
nopy, with an average of 0.593. In brief terms, these findings revealed a
low energy that is lost as fluorescence within the canopy and hence, a
high efficacy of the outside canopy in terms of photochemistry.
The results revealed a highly significant difference (p < 0.001) in
the electron transport rate (ETR) that was measured in different canopy
positions (Fig. 5a). The ETR was generally seven times higher (173 265)
in the leaves from the outside canopy, when compared to the ETR
measured in the leaves inside the canopy (24 124). Effective quantum
efficiency of PS II photochemistry (ϕPS II) was also found to vary sig-
nificantly (p < 0.001) with the canopy positions. The ϕPS II was re-
corded to be higher outside the canopy, with an average of 0.32, and
lower inside the canopy, with an average of 0.044 (Fig. 5b).
Photochemical quenching (qP) and the proportion of “open” PS II
reaction centres (1-qP) differed significantly (p < 0.001) with canopy
positions (Fig. 5c and d). The qP was generally higher outside canopy
S. Shezi, et al.
Fig. 3. Intrinsic water use efficiency (WUEi) (a) and linear electron transport rate (LETR) (b) of leaves in positions outside and within the canopy of ‘Gem’ avocado
cultivar. Values are means ± SE (n = 30).
Fig. 4. Maximum quantum efficiency of photosystem II photochemistry (ϕ PS
II) (d) of ‘Gem’ avocado leaves. Values are means ± SE (n = 30).
(0.52) and lower within the canopy (0.071). The 1-qP that was recorded
outside the canopy was approximately two times higher inside the ca-
nopy (0.92) than outside (0.48) (Fig. S3).
A highly significant difference (p < 0.001) was found in the thermal
heat dissipation (D) that was measured in different canopy positions
(Fig. 6a); it was slightly higher in the leaves from the inside canopy
(0.40) when compared to the outside canopy (0.37). A fraction of
photon energy absorbed by PS II antennae that was trapped by “open”
PS II reaction centres, to be used for photochemistry P also varied
significantly (p < 0.001) with canopy positions. The P was recorded to
be five times higher within the canopy, with an average of 0.52, and
lower outside the canopy, with an average of 0.10 (Fig. 6b).
Fig. 5. Electron transport rate (ETR) (a); quantum efficiency of photosystem II photochemistry (ϕ PS II) (b); and photochemical quenching (qP) (c) on ‘Gem’ avocado
leaves. Values are means ± SE (n = 30).
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Journal of Plant Physiology 246–247 (2020) 153130
The rate of heat dissipation (RHD) differed significantly (p < 0.001)
with canopy positions (Fig. 6c). The RHD measured within the canopy
was higher (326.1 photons m−2s-1) than the rate measured in leaves
from outside the canopy (299.5 photons m−2s-1). The rate of photo-
chemistry (RPC) was also found to vary significantly (p < 0.001) with
the canopy positions. The RPC was recorded to be seven times higher
within the canopy, having an average of 442.4 photons m−2s-1, and
lower outside the canopy with an average of 63.6 photons m−2s-1
(Fig. 6d). Photoinhibition of photosystem II reaction centres (Y (NO))
had highly significant variations (p < 0.001) between different canopy
positions (Fig. 7) and was generally higher within the canopy (0.96)
and lower outside the canopy (0.68).
3.2. Mesocarp dry matter content (DM)
The mesocarp dry matter content (DM) of avocado fruit varied
significantly (p < 0.001) with canopy position during fruit growth and
development (Fig. 8). The DM was higher in the fruit that were har-
vested outside the canopy, with an average of 28.9 %, and was lower in
fruit harvested within the canopy, averaging 26.9 %. A significant
variation (p < 0.001) in the fruit DM with sampling days (from 182 to
294 days after full bloom) was also observed. The DM was continuously
increasing over time as a sign of fruit development.
S. Shezi, et al. Journal of Plant Physiology 246–247 (2020) 153130

Fig. 6. Effect of canopy position on the D portion of the absorbed photon that was thermally dissipated (a); the fraction of photon energy trapped by “open” PS II
reaction centres and utilised for photochemistry (P) (b); rate of heat dissipation (RHD) (c); and the rate of photochemistry (RPC) (d). Values are means ± SE (n = 30).

Table 1
Pearson’s correlation coefficient (r) describing the correlations between pho-
tosynthetic parameters and DM within and outside the tree canopy.
Correlation coefficient (r) Within Correlation coefficient (r) Outside
canopy canopy

DM Significance test DM Significance test

A 0.53 * 0.58 *
gs 0.51 * 0.77 **
Ci 0.53 * 0.68 **
T 0.48 ns 0.68 **
Fo' −0.006 ns −0.24 ns
Fm' 0.009 ns −0.19 ns
Fv' 0.011 ns −0.086 ns
Fv'/Fm' −0.033 ns −0.16 ns
Fig. 7. Effect of canopy position on the portion of the absorbed photon that was ϕPSII 0.023 ns 0.15 ns
thermally dissipated (Y (NO)). Values are means ± SE (n = 30). qP 0.011 ns 0.086 ns
qN −0.026 ns −0.13 ns
ETR 0.48 ns 0.18 ns
1-qP −0.011 ns 0.086 ns
Y −0.023 ns −0.15 ns
P −0.033 ns 0.012 ns
D 0.026 ns −0.15 ns
RPC 0.026 ns 0.013 ns
RHD −0.033 ns −0.16 ns
LETR 0.023 ns 0.15 ns
WUEi 0.53 * 0.69 **
WUEins 0.53 * 0.67 **
A/Ci 0.12 ns 0.45 ns

A, photosynthesis; gs, stomatal conductance; T, transpiration rate; Fv’/Fm’,

Fig. 8. Effect of canopy position on mesocarp dry matter (DM) content (%)
during fruit development.
3.3. Correlations between photosynthetic parameters and DM in positions
within and outside the canopy
The correlation coefficients (r) between photosynthetic parameters
and the DM of fruit harvested in positions within and outside the ca-
nopy are presented in Table 1. The A, gs, Ci, A/gs and A/T were strongly
positive and significantly correlated with the DM. The strength of
maximum quantum yield of PS II photochemistry; ϕPS II, effective quantum
efficiency of photosystem II; qP, photochemical quenching; qN, non-photo-
chemical quenching; ETR, electron transport rate; 1-qp, proportion of open
reaction centers; D, thermal heat dissipation; RHD, rate of thermal heat dis-
sipation; P, fraction of photon energy absorbed by PS II antennae that was
trapped by “open” PS II reaction centres; RPC, rate of photochemistry; Y (NO),
photoinhibition of photosystem II reaction centres; A/Ci, ratio of photosynth-
esis and internal CO2; WUEi, intrinsic water use efficiency; and WUEins, the
instantaneous water use efficiency.
* Significant at 5 % level of significance.
** Significant at 1 % level of significance; ns = not significant.

6
S. Shezi, et al.
correlations was stronger outside the canopy than within the canopy.
Within the canopy, A, gs, Ci, A/gs and A/T were strongly positive and
significantly correlated (p < 0.001) with the DM. Outside the canopy,
A, gs, Ci, A/gs and A/T were also strongly positive and significantly
correlated (p < 0.001) with DM.
Several photosynthetic parameters such as ΦPS II, qP, ETR, and gas
exchange parameters such as gs, Ci, T, WUEi and WUEins with weak
correlation to DM, made an indirect contribution to accumulation of
DM by being positively correlated with A (data not shown).
Photoprotection parameters such as D, P, Y, RHD and RPC had a poor
correlation (r < 0.5) with DM, but had an indirect impact by being
positively correlated to non–photochemical quenching (qN). The
chlorophyll fluorescence parameters such as Fo’, Fm’, Fv’, Fv’/Fm’ and 1-
qP, which showed a weak correlation (r < 0.5) with DM, had a negative
indirect impact on DM by negatively correlating with A (data not
shown). The indirect negative impact on DM deduced from a negative
correlation of Fo’, Fm’, Fv’, Fv’/Fm’ and 1-qP with A, occurs because the
energy that empowers chlorophyll fluorescence is partitioned from the
total light energy that could be enhancing photochemistry. Therefore,
under high chlorophyll fluorescence, the A is reduced, resulting in the
low accumulation of DM.
4. Discussion
Plants require sunlight as a source of energy for photosynthesis;
however, absorption of excess sunlight affects functioning of PS I and
PS II, resulting in reduced CO2 assimilation (Takahashi and Badger,
2011). The lower rate of assimilation affects availability of carbohy-
drates necessary for fruit growth, development and maturity
(Bartholomew and Sanewski, 2018). Leaf canopy position affects pho-
tosynthetic capacity, due to the differences in absorption and quality of
light required for carbohydrate metabolism (Scartazza et al., 2016;
Chen et al., 2017). Furthermore, intercepted light energy is partitioned
to photochemistry and photoprotection, causing variation in the net
assimilation in positions within and outside the canopy. Variation in the
net photosynthesis has been linked with uneven maturity, which yields
uneven ripening of avocado fruit harvested from positions within and
outside the canopy.
In the current study, photosynthetic efficiency differed significantly
(p < 0.001) in positions within and outside the canopy, with leaves
from outside the canopy recording A rates of 29.5 mol CO2 m−2s-1
compared to 19.3 mol CO2 m−2s-1 that was measured within the ca-
nopy. Furthermore, a positive and significant (p < .0001) correlation
was observed between A and DM in both canopy positions, with a
correlation coefficient (r) of 0.53 and 0.58 of positions within and
outside canopy, respectively. The higher A measured outside the ca-
nopy was linked to the high light interception due to the direct sunlight
exposure (Ferraz et al., 2016; Shezi et al., 2019). In addition to the high
light interception, the source-sink strength tends to be stronger in the
fruit that are in the outside canopy, mainly because they are highly
likely to be supported by the leaves that are outside the canopy due to
their close proximity (Liu et al., 1999). Being closer to the leaves that
are highly photosynthesizing triggers the fruit from the outside canopy
to demand even more assimilates from the actively photosynthesizing
leaves, causing them to receive more assimilates than fruit within the
canopy (Liu et al., 1999; Shezi et al., 2019). Therefore higher A outside
the canopy suggests that fruit from this position receive more assim-
ilates than fruit from the inside canopy, and thus have the potential to
mature earlier than the ones within the canopy (Shezi et al., 2019).
Photosynthetic parameters such as ΦPS II, qP, ETR, and LETR (part
of ETR) contributed positively towards accumulation of DM by ensuring
high A. The rate of occurrence of these parameters was higher outside
the canopy, which resulted in the overall A being higher than within the
canopy. This could be associated with the optimized light interception
that occurs due to full exposure of the leaves to the sunlight. Uneven
assimilation in positions within and outside the canopy result in the
7
Journal of Plant Physiology 246–247 (2020) 153130
uneven overall distribution of assimilates to all sinks, which results in
an uneven fruit growth and development, and hence, maturity.
Distribution of assimilates is biased by the sink strength, which favours
fruit at an advanced stage of development.
Gas exchange parameters such as gs, Ci, WUEi and WUEins had a
stronger significant (p < 0.001) correlation with DM outside the canopy
than within. Stomatal conductance (gs), which defines the rate of CO2
entrance through stomata, was higher outside the canopy, resulting in
the higher Ci. The WUEi was driven by A and gs which were higher
outside the canopy, while WUEins was driven by T and A, which were
also higher outside than within the canopy. Both WUEi and WUEins were
higher within and lower outside the canopy (Fig. 3a and S1B), which
means leaves within the canopy were efficient in terms of using water.
This could be due primarily to the cooler temperature in the sur-
roundings of the leaves within the canopy, compared to exposed leaves
from outside canopy. Furthermore, morphological characteristics of the
leaves from the inside canopy have a large surface area, ensuring that
intercepted light is used efficiently in photochemistry.
Chlorophyll fluorescence parameters such as Fo’, Fm’, Fv’, Fv’/Fm’,
and 1-qP, which showed a weak correlation (r < 0.5) with DM, also had
a negative indirect impact on DM by negatively correlating with A (data
not shown). Minimum fluorescence occurs when all the antenna pig-
ment complexes associated with the photosystem are open (Hazrati
et al., 2016; Mashilo et al., 2018). The Fm’ denotes the maximum en-
ergy loss (fluorescence) that occurs when there is a maximum light
intensity flash applied (Maxwell and Johnson, 2000). Therefore, a
higher Fm’ inside the canopy in the current study means there was a
high chance of energy loss within the canopy compared to outside the
canopy when there was excess light exposure. This is due to the mor-
phological characteristics of the leaves from the inside canopy, which
are totally different from the ones from the outside canopy in the way
that they cannot intercept high light intensity. The leaves from the
outside canopy have thicker mesophyll cells and two to three layers of
palisade cells, which make them more efficient in maximizing light
interception (Chartzoulakis et al., 2002). The leaves from the inside
canopy have only a single layer of palisade cells. Therefore, in the
current study, higher Fm’ measured inside the canopy indicates a higher
possibility of causing photooxidative damage if the same leaves were to
be fully exposed to the sunlight.
The Fv’/Fm’ was measured as an indirect indicator of photoinhibi-
tion and it varied significantly with canopy positions. The Fv’/Fm’
measures the maximum quantum efficiency of the photosystem II
photochemistry. A high Fv’/Fm’ means the leaves have a strong pro-
tective mechanism of PS II against photoinhibition (Maxwell and
Johnson, 2000; Lichtenthaler et al., 2005; Klughammer and Schreiber,
2008). Therefore, in the current study, higher Fv’/Fm’ outside the ca-
nopy than inside it suggests that the leaves from the outside canopy
have a strong protective mechanism against photoinhibition of PS II
and possible photooxidative damage. A strong photoprotection occurs,
due to higher non-photochemical quenching (NPQ) of the chlorophyll
fluorescence, commonly phrased as heat dissipation (D). The higher
rate of heat dissipation (RHD) in the leaves from the outside canopy
could be due to the de-epoxidation state (DEPS) of the xanthophyll
cycle pigments, which is known to be higher in leaves that are under
high light intensity. Filella et al. (2009) reported that the DEPS and V +
A+Z (violaxanthin + antheraxanthin + zeaxanthin)/chlorophyll- a +
b of Scots pine (Pinus sylvestris L.) and Holm oak (Quercus ilex L.) is
higher in the leaves that are fully exposed to the sun than the shade
leaves. This suggests that the DEPS and V + A+Z have been higher in
the current study. The higher de-epoxidation of the xanthophyll cycle in
the leaves from the outside canopy was also linked to the higher β-
carotene and total carotenoid contents.
Therefore, in the current study, it could also be hypothesized that a
stronger photoprotection mechanism of the leaves from the outside
canopy is due to higher β-carotene and total carotenoid content than in
the leaves from the inside canopy. Yahia et al. (2017) reported the
S. Shezi, et al.
importance of light in biosynthesis of carotenoids, particularly zeax-
anthin, β-carotene, α-carotene, and α-cryptoxanthin in avocado meso-
carp tissue. This could be one of the reasons why the overall photo-
synthetic efficiency was higher in the outside canopy.
The lutein epoxide cycle (Lx cycle), which is based on inter-
conversions of lutein-5,6-epoxide (Lx) and lutein (L) synthesized from
α-carotene (α-C), is known to operate in avocado and other terrestrial
plants in response to excess light. The high concentration of α-carotene
synthesized in the leaves from the outside canopy suggests higher oc-
currences of the Lx cycle in that position than inside the canopy.
Occurrence of the Lx cycle is necessary for the conversion of Lx to L.
Lutein-5,6-epoxide has been known to increase over time in the leaves
from the inside canopy (shade leaves), with its capacity to enhance light
harvesting efficiency. Lutein has been reported to be a primary re-
spondent that is photoprotective, and hence it declines in response to
exposure of leaves to high light intensity (Förster et al., 2009). Johnson
et al. (2007) also reported that L could be the earliest and most active
respondent against photooxidative damage in Arabidopsis thaliana
plants. Peng et al. (2006) reported that L is a credible alternative to Z as
an operative scavenger of the reactive oxygen species (ROS) in vitro.
Possibly, L chemical quenching could have a role as an early target for
photooxidation in the antenna or reaction centres of the photosynthesis
apparatus, as does the D1 reaction centre protein of PS II that is the
most vulnerable to photooxidative damage, thus protecting other PS II
proteins (Chow et al., 2005). It has been suggested that the lower
photosynthetic efficiency under shade could be associated with the
photoconverted L that “locks in” the photoprotective energy dissipation
in the shade (Förster et al., 2009). Therefore, there is less non-photo-
chemical quenching or energy lost by thermal dissipation in the pre-
sence of high Lx, which emphasizes less photoprotection in matured
avocado leaves from the inside canopy. A strong protective mechanism
of the leaves from the outside canopy could also be associated with the
unique morphological features such as thicker mesophyll cells and two
to three layers of palisade cells, which make them more efficient in
maximizing light interception (Chartzoulakis et al., 2002).
Photoinhibition of PS II reaction centres (Y) was recorded to be
significantly (p < 0.001) higher within the canopy and lower outside
the canopy. The Y measures the photoinhibition of PS II reaction cen-
tres, which defines the inability of a plant to protect itself from damage
from the excess light energy (Mashilo et al., 2018). The P was sig-
nificantly higher (p < 0.001) in the inside canopy and lower outside the
canopy. The P measures the fraction of photon energy absorbed by PS II
antennae that is trapped by “open’’ PS II reaction centres (centres with
8
Journal of Plant Physiology 246–247 (2020) 153130
Fig. 9. Principal component biplot presenting PC dimension 1
vs PC dimension 2 of grouped photoprotection and photo-
synthesis parameters, as well as DM measured within and
outside canopy positions, photosynthesis (A), stomatal con-
ductance (gs), thermal heat dissipation (D), rate of thermal
heat dissipation (RHD), fraction of photon energy absorbed by
PS II antennae that was trapped by “open” PS II reaction
centres (P), rate of photochemistry (RPC), photoinhibition of
photosystem II reaction centres (Y (NO)), intrinsic water use
efficiency (WUEi), and ratio of photosynthesis and internal
CO2 (A/Ci).
QA in the oxidized state) and utilized in PS II photochemistry (Maxwell
and Johnson, 2000). The higher P within the canopy in the current
study implies that there was more photon energy that was trapped in
the open reaction centres within the canopy, when compared to the
outside canopy. The high P measured within the canopy could be linked
with the high proportion of open reaction centres measured inside the
canopy, compared to outside it.
The D that was measured inside the canopy was significantly higher
(p < 0.001) than the one measured outside it. The D measures the
portion of absorbed photon energy that is thermally dissipated
(Maxwell and Johnson, 2000; Mashilo et al., 2018). The thermal dis-
sipation of photon energy is one of the photoprotective mechanisms
against excess light (Guidi et al., 2017). The RHD varied significantly
(p < 0.001) with the canopy positions. It was generally higher inside
and lower outside the canopy. The RHD measures the rate of heat
dissipation. The rate of photochemistry (RPC), which measures the rate
of photochemistry (Maxwell and Johnson, 2000; Mashilo et al., 2018),
varied significantly (p < 0.001) with canopy position; i.e. RPC was
measured higher inside and lower outside the canopy.
Photoprotection parameters such as D, P, Y, RHD and RPC had a
poor correlation (r < 0.5) with DM, but had an indirect impact by being
positively correlated to non–photochemical quenching, (qN). The
overall performance of photoprotection mechanism parameters Y, D,
RHD, P and RPC varied significantly (p < 0.001) with canopy position.
All of the photoprotection parameters were higher within the canopy
and lower outside it (Fig. 9). This means that the great proportion of
accumulated energy was used for protection against excess light. This
conversely means less energy availability for photochemistry, which is
why the overall A was lower within the canopy and higher outside it.
The energy that was accumulated inside the canopy was sufficient for
maintaining the balance between the two major activities, i.e. to
maintain photochemistry and to protect against excess light (Maxwell
and Johnson, 2000). This means ‘Gem’ avocado trees have a high ca-
pacity to compensate for both photosynthetic efficiency and photo-
protection. This high compensatory efficacy of ‘Gem’ was measured by
the yield and quality of fruit not being compromised by different ca-
nopy positions. However, fruit that were harvested within the canopy
had a two-week delay in reaching the minimum maturity (25 % DM),
which had already been reached by fruit harvested from the outside
canopy. The present findings suggest that reduced photosynthetic effi-
ciency of ‘Gem’ avocado fruit harvested from within the canopy does
not compromise fruit DM. However, harvesting fruit from within the
canopy at least two weeks after those from outside the canopy can be
S. Shezi, et al.
advantageous.
The results revealed a significant difference (p < 0.001) in the
mesocarp dry matter content (DM) between fruit harvested inside or
outside the canopy. Fruit that were harvested outside the canopy had
higher accumulated DM over time, and eventually matured earlier than
fruit harvested within the canopy. The high accumulation of DM in the
fruit harvested from the outside canopy was positively correlated with
the overall rate of A, which was high in the leaves oriented in the
outside canopy. Lahav and Trochoulias (1982) also reported a high
accumulation of DM in ‘Hass’ avocados that were fully exposed to the
sun. The higher A rate in the leaves from the outside canopy was due to
the optimized light interception, while accumulation of more assim-
ilates from these leaves was attributed to the sink strength, since fruit
from this outside position attracts more assimilates due to strong sink
strength as well as proximity to highly photosynthesizing leaves (Liu
et al., 1999). The overall findings revealed that the fruit from the
outside canopy have the potential of maturing earlier that those from
the inside canopy. The leaves from the inside were able to compensate
for photoprotection without compromising the accumulation of DM,
but a significant difference in the DM could result in an uneven ripening
if fruit from both canopies are harvested at the same time (Shezi et al.,
2019).
5. Conclusion
The overall findings for this study revealed the fact that the effi-
ciency of photochemistry and photoprotection are concomitant with the
light intensity. Under low light intensity there is high efficiency of
photochemistry and optimum photoprotection. The efficiency of pho-
tochemistry is because under low light intensity, the utilized light en-
ergy is almost equivalent to the absorbed or intercepted light energy.
Optimum photoprotection occurs because under low light intensity, the
excess energy (difference between absorbed and utilized energy) is at a
minimum; under high light intensity, the excess energy is at a max-
imum. An excess of energy has the huge potential of causing photo-
oxidative damage to the photosynthetic membrane (Ruban et al.,
2012). In the current study, the leaves within the canopy showed a low
overall photosynthetic efficiency and a high photoprotective me-
chanism against excess light, while the leaves from outside the canopy
showed a vice versa effect. The low efficiency of photochemistry in the
leaves from within the canopy was due to low light interception oc-
curring, since most of the leaves were shaded. The high photoprotection
was due to the fact that most of the absorbed light was utilized in
photochemistry, i.e. the excess energy was at a minimum. The high
efficiency of photochemistry was due to maximized light interception
because of direct exposure to the light. However, the photoprotection
was too low, mainly because the excess energy was at a maximum. The
overall findings revealed that the fruit from the outside canopy have a
potential to mature earlier that the ones within canopy. This was linked
to the overall leaf A and DM, which were recorded higher outside the
canopy. The leaves within the canopy were able to compensate for
photoprotection without compromising the accumulation of DM, and
hence maturity; however, a significant difference in the DM can result
in an uneven ripening if fruit from both canopies are harvested at the
same time. Therefore, the application of these findings is that the South
African avocado industry can delay harvesting fruit within the canopy
by at least two weeks as a way of enabling them to accumulate more
assimilates and increase their DM while continuing the marketing of
early matured fruit from the outside canopy. This may help in ex-
tending the marketing season, while at the same time raising the DM of
fruit within the canopy to the acceptable minimum DM for harvesting.
CRediT authorship contribution statement
Sabelo Shezi: Investigation, Formal analysis, Visualization, Writing
- original draft. Lembe Samukelo Magwaza: Funding acquisition,
9
Journal of Plant Physiology 246–247 (2020) 153130
Supervision, Conceptualization, Resources, Writing - review & editing.
Jacob Mashilo: Data curation, Writing - review & editing. Samson
Zeray Tesfay: Supervision, Writing - review & editing. Asanda
Mditshwa: Supervision, Writing - review & editing.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgements
The College of Agriculture, Engineering and Science of the
University of KwaZulu-Natal (UKZN) and the National Research
Foundation (NRF) of South Africa are acknowledged for their financial
support of this study. Westfalia Fruit in Howick, KwaZulu-Natal, South
Africa, is also acknowledged for providing the orchard and the fruit for
this experiment.
Appendix A. Supplementary data
Supplementary material related to this article can be found, in the
online version, at doi:https://doi.org/10.1016/j.jplph.2020.153130.
References
Araus, J., Amaro, T., Voltas, J., Nakkoul, H., Nachit, M., 1998. Chlorophyll fluorescence
as a selection criterion for grain yield in durum wheat under Mediterranean condi-
tions. Field Crops Res. 55, 209–223. https://doi.org/10.1016/S0378-4290(97)
00079-8.
Bartholomew, D.P., Sanewski, G.M., 2018. 11 inflorescence and fruit development and
yield. Pineapple: Bot. Prod. Uses 233. https://doi.org/10.1079/9781786393302.
0233.
Bertling, I., Cowan, A., 1998. Effect of Photo-Inhibition on Fruit Growth and Development
in Hass Avocado 21. South African Avocado Growers’ Association Yearbook, pp.
36–38 http://www.avocadosource.com/journals/saaga/saaga_1998/saa-
ga_1998_pg_036-038.pdf.
Chartzoulakis, K., Patakas, A., Kofidis, G., Bosabalidis, A., Nastou, A., 2002. Water stress
affects leaf anatomy, gas exchange, water relations and growth of two avocado cul-
tivars. Sci. Hortic. 95, 39–50. https://doi.org/10.1016/S0304-4238(02)00016-X.
Chen, B., Yang, H., Ma, Y., Liu, J., Lv, F., Chen, J., Meng, Y., Wang, Y., Zhou, Z., 2017.
Effect of shading on yield, fiber quality and physiological characteristics of cotton
subtending leaves on different fruiting positions. Photosynthetica 55, 240–250.
https://doi.org/10.1007/s11099-016-0209-7.
Chow, W.S., Lee, H.Y., He, J., Hendrickson, L., Hong, Y.N., Matsubara, S., 2005.
Photoinactivation of photosystem II in leaves. Photosynth. Res. 84, 35–41. https://
doi.org/10.1007/s11120-005-0410-1.
Demmig-Adams, B., Adams Iii, W., 1992. Photoprotection and other responses of plants to
high light stress. Annu. Rev. Plant Biol. 43, 599–626. https://doi.org/10.1146/
annurev.pp.43.060192.003123.
Ferraz, T.M., Rodrigues, W.P., Netto, A.T., De Oliveira Reis, F., Pecanha, A.L., De Assis,
F.M., De Sousa, E.F., Glenn, D.M., Campostrini, E., 2016. Comparison between single-
leaf and whole-canopy gas exchange measurements in papaya (Carica papaya L.)
plants. Sci. Hortic. 209, 73–78. https://doi.org/10.1016/j.scienta.2016.06.014.
Filella, I., Porcar-Castell, A., Munné-Bosch, S., Bäck, J., Garbulsky, M.F., Peñuelas, J.,
2009. PRI assessment of long-term changes in carotenoids/chlorophyll ratio and
shortterm changes in de-epoxidation state of the xanthophyll cycle. Int. J. Remote
Sens. 30 (17), 4443–4455. https://doi.org/10.1080/01431160802575661.
Fischer, G., Almanza-Merchán, P.J., Ramírez, F., 2012. Source-sink relationships in fruit
species: a review. Rev. Colomb. Cienc. Hortíc. 6, 238–253.
Flexas, J., Escalona, J., Medrano, H., 1999. Water stress induces different levels of pho-
tosynthesis and electron transport rate regulation in grapevines. Plant Cell Environ.
22, 39–48. https://doi.org/10.1046/j.1365-3040.1999.00371.x.
Förster, B., Osmond, C.B., Pogson, B.J., 2009. De novo synthesis and degradation of lx and
V cycle pigments during shade and sun acclimation in avocado leaves. Plant Physiol.
149, 1179–1195. https://doi.org/10.1104/pp.108.131417.
Guidi, L., Tattini, M., Landi, M., 2017. How does chloroplast protect chlorophyll against
excessive light. Chlorophyll 21. https://doi.org/10.5772/67887.
Hakala, M., Tuominen, I., Keränen, M., Tyystjärvi, T., Tyystjärvi, E., 2005. Evidence for
the role of the oxygen-evolving manganese complex in photoinhibition of photo-
system II. BBA-Bioenergetics 1706, 68–80. https://doi.org/10.1016/j.bbabio.2004.
09.001.
Hazrati, S., Tahmasebi-Sarvestani, Z., Modarres-Sanavy, S.a.M., Mokhtassi-Bidgoli, A.,
Nicola, S., 2016. Effects of water stress and light intensity on chlorophyll fluorescence
parameters and pigments of Aloe vera L. Plant Physiol. Biochem. 106, 141–148.
https://doi.org/10.1016/j.plaphy.2016.04.046.
S. Shezi, et al.
Johnson, M.P., Havaux, M., Triantaphylidès, C., Ksas, B., Pascal, A.A., Robert, B.,
Davison, P.A., Ruban, A.V., Horton, P., 2007. Elevated zeaxanthin bound to oligo-
meric lhcii enhances the resistance of Arabidopsis to photooxidative stress by a lipid-
protective, antioxidant mechanism. J. Biol. Chem. 282, 22605–22618. https://doi.
org/10.1074/jbc.M702831200.
Klughammer, C., Schreiber, U., 2008. Complementary ps ii quantum yields calculated
from simple fluorescence parameters measured by Pam Fluorometry and the sa-
turation pulse method. PAM Appl. Notes 1, 201–247. https://doi.org/10.1007/
s11099-005-0062-6.
Lahav, E., Trochoulias, T., 1982. The effect of temperature on growth and dry matter
production of avocado plants. Aust. J. Agric. 33, 549–558. https://doi.org/10.1071/
AR9820549.
Lichtenthaler, H., Buschmann, C., Knapp, M., 2005. How to correctly determine the dif-
ferent chlorophyll fluorescence parameters and the chlorophyll fluorescence decrease
ratio r fd of leaves with the pam fluorometer. Photosynthetica 43, 379–393. https://
doi.org/10.1007/s11099-005-0062-6.
Liu, X., Robinson, P.W., Madore, M.A., Witney, G.W., Arpaia, M.L., 1999. Hass’ avocado
carbohydrate fluctuations. II. Fruit growth and ripening. J. Am. Soc. Hortic. Sci. 124,
676–681. https://doi.org/10.21273/JASHS.124.6.676.
Magwaza, L.S., Tesfay, S.Z., 2015. A review of destructive and non-destructive methods
for determining Avocado fruit maturity. Food Bioprocess Technol. 8, 1995–2011.
https://doi.org/10.1007/s11947-015-1568-y.
Martin, B., Ruiz-Torres, N.A., 1992. Effects of water-deficit stress on photosynthesis, its
components and component limitations, and on water use efficiency in wheat
(Triticum aestivum L.). Plant Physiol. 100, 733–739. https://doi.org/10.1104/pp.100.
2.733.
Mashilo, J., Shimelis, H., Odindo, A., 2017. Yield-based selection indices for drought
tolerance evaluation in selected bottle gourd [Lagenaria siceraria (Molina) Standl.]
landraces. Acta Agric. Scand. B—S P 67, 43–50. https://doi.org/10.1080/09064710.
2016.1215518.
Mashilo, J., Odindo, A., Shimelis, H., Musenge, P.A., Tesfay, S.Z., Magwaza, L.S., 2018.
Photosynthetic efficiency of bottle gourd [Lagenaria siceraria (Molina) Standl.] under
drought stress. Indian J. Plant Physiol. 23, 293–304. https://doi.org/10.1007/
s40502-018-0377-5.
Maxwell, K., Johnson, G.N., 2000. Chlorophyll fluorescence—A practical guide. J. Exp.
Bot. 51, 659–668. https://doi.org/10.1093/jexbot/51.345.659.
Ncama, K., Magwaza, L.S., Poblete-Echeverría, C.A., Nieuwoudt, H.H., Tesfay, S.Z.,
Mditshwa, A., 2018. On-tree indexing of ‘Hass’ avocado fruit by non-destructive as-
sessment of pulp dry matter and oil content. Biosyst. Eng. 174, 41–49. https://doi.
org/10.1016/j.biosystemseng.2018.06.011.
Oguchi, R., Onoda, Y., Terashima, I., Tholen, D., 2018. Leaf Anatomy and Function. The
Leaf: A Platform for Performing Photosynthesis. Springerhttps://doi.org/10.1086/
335974.
10
Journal of Plant Physiology 246–247 (2020) 153130
Olarewaju, O.O., Bertling, I., Magwaza, L.S., 2016. Non-destructive evaluation of avocado
fruit maturity using near infrared spectroscopy and PLS regression models. Sci.
Hortic. 199, 229–236. https://doi.org/10.1016/j.scienta.2015.12.047.
Peng, C.-L., Lin, Z.-F., Su, Y.-Z., Lin, G.-Z., Dou, H.-Y., Zhao, C.-X., 2006. The antioxidative
function of lutein: electron spin resonance studies and chemical detection. Funct.
Plant Biol. 33, 839–846. https://doi.org/10.1071/FP06013.
Powles, S.B., 1984. Photoinhibition of photosynthesis induced by visible light. Annu. Rev.
Plant Physiol. 35, 15–44. https://doi.org/10.1146/annurev.pp.35.060184.000311.
Ruban, A.V., Johnson, M.P., Duffy, C.D., 2012. The photoprotective molecular switch in
the photosystem II antenna. Acta Agric. Scand. B—S P 1817, 167–181. https://doi.
org/10.1016/j.bbabio.2011.04.007.
Scartazza, A., Di Baccio, D., Bertolotto, P., Gavrichkova, O., Matteucci, G., 2016.
Investigating the European beech (Fagus sylvatica L.) leaf characteristics along the
vertical canopy profile: leaf structure, photosynthetic capacity, light energy dissipa-
tion and photoprotection mechanisms. Tree Physiol. 36, 1060–1076. https://doi.org/
10.1093/treephys/tpw038.
Shezi, S., Magwaza, L.S., Mashilo, J., Tesfay, S.Z., Mditshwa, A., 2019. Photosynthetic
efficiency and relationship to mesocarp dry matter content of ‘Carmen’avocado
(Persea americana Mill.) fruit in a cool subtropical climate. Sci. Hortic. 253, 209–216.
https://doi.org/10.1016/j.scienta.2019.04.048.
Takahashi, S., Badger, M., 2011. Photoprotection in plants: a new light on photosystem II
damage. Trends Plant Sci. 16, 53–60. https://doi.org/10.1016/j.tplants.2010.10.001.
Whiley, A., Schaffer, B., Lara, S., 1992. Carbon dioxide exchange of developing avocado
(Persea americana Mill.) fruit. Tree Physiol. 11, 85–94. https://doi.org/10.1093/
treephys/11.1.85.
Woolf, A., Ferguson, I., Requejo-Tapia, L., Boyd, L., Laing, W., White, A., 1999. Impact of
sun exposure on harvest quality of ‘Hass’ avocado fruit. Rev. Chapingo Ser. Hortic. 5,
352–358. http://www.avocadosource.com/WAC4/WAC4_p353.pdf.
Woolf, A., Clark, C., Terander, E., Phetsomphou, V., Hofshi, R., Arpaia, M.L., Boreham, D.,
Wong, M., White, A., 2003. Measuring avocado maturity; ongoing developments.
Orchidardist 40–45. http://209.143.153.251/Journals/Orchardist/
WoolfAllan2003b.pdf.
Yahia, E.M., De Jesús Ornelas-Paz, J., Emanuelli, T., Jacob-Lopes, E., Zepka, L.Q.,
Cervantes-Paz, B., 2017. Chemistry, Stability, and Biological Actions of Carotenoids.
Fruit and Vegetable Phytochemicals: Chemistry and Human Health 2. pp. 285–346.
https://doi.org/10.1002/9781119158042.ch15.
Zeng, G., Guo, Y., Xu, J., Hu, M., Zheng, J., Wu, Z., 2017. Partial shade optimizes pho-
tosynthesis and growth in bayberry (Myrica rubra) trees. Hortic. Environ. Biotechnol.
58, 203–211. https://doi.org/10.1007/s13580-017-0003-x.
Zsiros, O., Allakhverdiev, S.I., Higashi, S., Watanabe, M., Nishiyama, Y., Murata, N.,
2006. Very strong UV-A light temporally separates the photoinhibition of photo-
system II into light-induced inactivation and repair. BBA Bioenergetics 1757,
123–129. https://doi.org/10.1016/j.bbabio.2006.01.004.