Keywords
Photorespiration, high temperature, high-light, alternative electron
acceptors, cowpea, , heat stress, net CO2 assimilation,
PSII efficiency
INTRODUCTION
Heat stress is a major environmental challenge for plants, and it is
well established that, in a wide range of species, high temperature (HT)
stress can lead to declines in photosynthesis (Cui, Li, Fan, Xu, &
Zhang, 2006; Panigrahi, Pradhan, Panda, Panda, & Joshi, 2016; Tan,
Meng, Brestic, Olsovska, & Yang, 2011; Wise, Olson, Schrader, &
Sharkey, 2004). The impact of HT stress on photosynthesis is dependent
on factors such as leaf developmental age, duration and previous history
of exposure (Hasanuzzaman, Nahar, Alam, Roychowdhury, & Fujita, 2013),
as well as other environmental factors, especially high-light (HL)
stress, which often co-occur with and can be compounded by HT (Weis,
1982).
While light energy is essential for photosynthesis, excessive light can
cause detrimental effects on photosynthesis, a phenomenon known as
photoinhibition or photodamage (Aro, Virgin, & Andersson, 1993; Long,
Humphries, & Falkowski, 1994; Nishiyama & Murata, 2014). HL stress
causes reductions in photosynthesis, especially under rapidly
fluctuating environmental conditions, which ultimately results in loss
of crop productivity (Slattery, Walker, Weber, & Ort, 2018). Plants
have evolved various mechanisms to deal with excessive light and prevent
damage, including changes in leaf orientation (Gamon & Pearcy, 1989;
Pastenes, Pimentel, & Lillo, 2005), chloroplast movement (Suetsugu,
Higa, Gotoh, & Wada, 2016; Wada, 2013) and non-photochemical quenching
(NPQ) processes that dissipate excitation energy (Ruban, 2016; Stefanov
& Terashima, 2008). There are several forms of NPQ, including the
rapidly reversible “energy dependent” quenching
(q E), which is activated by acidification of the
thylakoid lumen (Avenson, Cruz, & Kramer, 2004; Li et al., 2004), and
more slowly-reversible forms including q I, which
is caused by accumulation of inactive (photodamaged) photosystem II
(PSII) centers (Krause, Somersalo, Zumbusch, Weyers, & Laasch, 1990)
and state transitions (q T) that can divert
PSII-associated light harvesting complexes (LHCs) to PSI. Depending on
its redox state, PSI can either use light energy from these LHCs for PSI
photochemistry, or dissipate it as heat (Butler, 1978; Joly &
Carpentier, 2007).
The importance of light for thermotolerance of photosynthesis has been
noted in previous work (Buchner, Stoll, Karadar, Kranner, & Neuner,
2015; Havaux, Greppin, & Strasser, 1991; Weis, 1982). Although, there
have been several studies that show that HL combined with HT accentuates
photoinhibition (e.g. Chen et al., 2017; Havaux, 1992; Lu et al., 2017),
there are also reports, for instance in tomatoes, where combined
moderate HT and HL improved tolerance of photosynthesis compared to
either treatment alone (Gerganova, Popova, Stanoeva, & Velitchkova,
2016). However, since different species have different light intensity
requirements (e.g shade vs. sun adapted plants, (Hemming, 2011; C. H.
Lin, McGraw, George, & Garret, 1999)), it is very likely that responses
to combinations of HT and HL stress are dependent on genotype, species
or acclimation during development. Nevertheless, several components of
the responses may be conserved among species.
During photosynthesis, light energy captured by the light harvesting
complex is used to drive electrons by linear electron flow (LEF) to
store reducing power in NADPH/O2 and phosphorylation
potential in ATP/ADP+Pi, which together drive the carbon assimilation
reactions in the Calvin-Benson-Bassham (CBB) cycle and other cellular
processes. In principle, both the light reactions and CBB cycle may be
affected by HT. For example, it is well-established that HT decreases
the specificity of rubisco and solubility of CO2relative to O2 (Brooks & Farquhar, 1985; Galmés,
Hermida-Carrera, Laanisto, & Niinemets, 2016), thus increasing
“photorespiratory pressure” or rates of photorespiration faster than
the rate of rubisco carboxylation (Schuster & Monson, 1990).
In addition to this shift from assimilation to photorespiration,
moderately high temperatures (35–40 °C) decrease rubisco activity,
which can be attributed to the loss of rubisco activase activity,
leading to progressive deactivation of rubisco by accumulation of
inhibitory products such as sugar phosphates (Crafts-Brandner &
Salvucci, 2000; Salvucci & Crafts-Brandner, 2004). While these studies
suggest that reduction in rubisco activity is a primary deleterious
effect of HT stress, it has also been proposed (Sharkey, 2005) that this
effect constitutes an adaptive mechanism possibly to avoid situations
where the damage by toxic products of photorespiration outweighs the
advantage of higher carbon fixation rates.
A rubisco activase feedback mechanism of this type could allow for
“safe” downregulation of assimilation. However, this mode of
regulation should decrease the availability of sinks for the products of
the light reactions. Without coordinated downregulation of the light
reactions, this mode of regulation could result in the accumulation of
reactive intermediates and subsequent photodamage. Indeed, it has been
proposed that high rates of photorespiration under environmental
stresses can serve a photoprotective role by maintaining electron
acceptor sinks (Huang, Hu, & Zhang, 2015; Voss, Sunil, Scheibe, &
Raghavendra, 2013). However, if rubisco is completely deactivated, rates
of photorespiration would also be decreased. Thus, many times HT results
in decreases in assimilation that are not fully accounted for by
increased photorespiration (Sharkey, 2005), suggesting that feedback
systems must operate to coordinate the capture of light in response to
these changes in assimilation, as suggested by observations that HT
induced decreases in assimilation in wheat were not accompanied by
increased PSII photodamage (Kalituho, Pshybytko, Kabashnikova, & Jahns,
2003).
The light and assimilatory reactions are tightly co-regulated to balance
the needs for efficient energy capture, while balancing energy storage
into ATP and NADPH to meet the needs of downstream metabolic reactions
and prevent the accumulation of reactive intermediates that can lead to
photodamage (Kramer & Evans, 2011; (Walker, Kramer, Fisher, & Fu,
2020). The thylakoid proton motive force (pmf ) plays a central
role in this co-regulatory network (Avenson et al., 2005; Kanazawa &
Kramer, 2002). The ΔpH component of pmf causes acidification of
the lumen, which activates q E (Li, Muller-Moule,
Gilmore, & Niyogi, 2002; Ruban, Johnson, & Duffy, 2012) and controls
electron flow at the level of the cytochromeb 6f complex, in a process known as
“photosynthetic control” (Tikhonov, 2014). The pmf and its
effects on downregulation are sensitive to processes that alter the
influx of protons such as LEF, cyclic electron flow (CEF) and the
conductivity of the thylakoid membrane to proton efflux
(g H+). The latter is governed
mainly by the chloroplast ATP synthase, and the partitioning ofpmf into its two components, Δψ and ΔpH. For example, decreasing
CO2 level slows assimilation and thus decreases LEF and
the flux of protons into the lumen. However, ATP synthase also slows, so
despite the fact that proton influx is slowed, the thylakoid builds up a
larger pmf , leading to activation of q Eand increased photosynthetic control (Kanazawa & Kramer, 2002).
Moderate HT has been shown to modulate this co-regulatory network in
tobacco by altering g H+, the
rates of violaxanthin de-epoxidase (VDE) and zeaxanthin epoxidase (ZE)
and the partitioning of pmf into Δψ and ΔpH (Zhang, Kramer, Cruz,
Struck, & Sharkey, 2011), but it is unclear how HL interacts with these
regulatory responses.
In this study, we focused on the effect of light on responses of
photosynthesis to HT in cowpea (Vigna unguiculata ) seedlings
having leaves of different maturity, with the aim of determining the
relative effects on assimilation and light reactions and their
co-regulation. We focused on cowpea because it is an agriculturally
important crop that serves as an essential source of livelihood and
nutrition for many in developing countries, many of which have hot
climates. Characterized by quick growth and broad leaves, it is ideal
for studying the interaction between HL and HT since it exhibits
considerable genetic variation in tolerance to these traits (Ehlers &
Hall, 1996, 1998).
Our data shows that in low light (LL), HT caused substantial decreases
in CO2 assimilation and PSII activity, while HT in
combination with HL led to a stimulation of PSII activity, but decreases
in CO2 assimilation compared to growth temperature (GT).
Further experiments revealed differences in the rates of PSII
photodamage between GT+HL and HT+HL, whereas PSII activity was not
enhanced in HT+HL under non-photorespiratory conditions, suggesting
photorespiration and other alternative electron acceptors to be
essential for tolerance of PSII to HT+HL stress.