TRPLSC 1615 No.
of Pages 3
Forum
Can Peroxisomes
Inform Cellular
Response to
Drought?
Andrei Smertenko1,*
Although plant germplasm con-
tains numerous unknown traits
for drought-tolerance, the ability
to phenotype this material
remains challenging. I propose
that peroxisome abundance may
be a viable parameter for the eval-
uation of drought response at the
cellular level, and hence could be
useful to improve phenotyping
efforts.
Strategies for Breeding Drought-
Tolerant Crops
The productivity of agricultural systems is
proportional to the amount of accessible
soil moisture [1]. Currently, water deficit
restricts yield in over one quarter of all
arable land worldwide [2]. Climate
change is predicted to reduce water
availability in many agricultural areas
where it is already a limiting factor. Apart
from reducing yield, drought also exac-
erbates the impact of other biotic and
abiotic stressors including soil salinity,
extremes in temperature, and pests/
pathogens, among others. While irriga-
tion can compensate for low rainfall, the
growing demand for water from industrial
and domestic users decreases the fea-
sibility of this option in many parts of the
world, particularly in those with arid cli-
mates. Traditionally, greater efficiency in
water use has been achieved through
optimizing water management and agri-
cultural practices [3]. Looking to the
future, further improvements in water
use efficiency will likely depend on the
development of new crop varieties with
the innate ability to sustain themselves in
a low-moisture environment.
Two approaches have been employed for
breeding more stress-tolerant crops [3].
First, the ‘bottom-up’ approach, relies on
modifications of individual genes that are
already known to be part of the tolerance
mechanism. The outcome of this
approach may not meet expectations
because yield is a genetically complex
trait controlled by a hierarchical, intercon-
nected network of multiple genes. A suc-
cessful outcome results from the
combined output of many interactions,
although the contribution of individual
genes and/or interactions could be rela-
tively small. Furthermore, the potential
effect from changing the activity of one
or several genes can be abrogated if other
members of the network behave differ-
ently due to the variability of seasonal and
geographic growth conditions, or as a
consequence of the trade-off effect on
the yield.
Second, the ‘top-down’ approach,
encompasses screening for desired traits
in a genetically diverse population. An
important advantage of this approach is
the potential to identify novel traits for
drought-tolerance in germplasm. How-
ever, phenotyping of large germplasm
collections for drought responses
remains challenging due to marked vari-
ability in (i) life cycle (e.g., early or late
flowering), (ii) morphology (e.g., root
architecture or plant size), and (iii) cellular
ultrastructure (e.g., cell wall thickness and
composition). This variability prevents the
normalized application of different stress-
ors to a specific life-cycle stage across all
accessions. Hence, the efficacy of ‘top-
down’ programs largely depends on a
battery of phenotyping tools that can sen-
sitively and selectively capture drought
responses on the population level. Cur-
rent tools used to phenotype plant mor-
phology, physiology, and yield [2] have
poor resolution for probing cellular
responses; however, these cellular
parameters could improve the sensitivity
and discrimination of phenotyping efforts
because they integrate the complexity of
diverse molecular interactions underlying
stress responses into a single readout.
Because peroxisomes play a key role in
stress responses, I hypothesize they are
the ideal candidate for such a cellular
parameter that could be used to improve
current phenotyping strategies.
Peroxisomes Alleviate Oxidative
Damage [90_TD$IF]under Stress
Peroxisomes are ubiquitous eukaryotic
organelles that are surrounded by a single
boundary membrane [4]. Among their
many roles are photorespiration (as part
of glycolate cycle), b-oxidation of fatty
acids, and the synthesis of hormones
indole-3-acetic acid and jasmonic acid
[4]. Moreover, the bulk of reactive oxygen
species (ROS) metabolism takes place in
peroxisomes [5]. The high concentration
of ROS exposes peroxisomal compo-
nents to risk from oxidative damage,
which is averted by ROS scavengers such
as (i) the antioxidants ascorbate and glu-
tathione (GSH) and (ii) a diverse cadre of
enzymes including catalase (CAT),
[92_TD$IF]monodehydroascorbate reductase,
GSH reductase, ascorbate peroxidase,
peroxiredoxins, and superoxide dismut-
ase (SOD).
These peroxisomal components play a
key role in several avenues of the stress
response. For example, the activity of
SOD and ascorbate peroxidase increases
during drought in peas [6], and drought-
tolerant rice varieties consistently contain
less H2O2 and exhibit higher activities of
CAT, SOD, and ascorbate peroxidase [7].
In addition to upregulation of ROS scav-
engers/enzymes, the total number of per-
oxisomes in cells can increase during
abiotic stress ([8] and citations therein).
This increase is accompanied by upregu-
lation of peroxisome biogenesis genes in
Arabidopsis, including PEX1, PEX10, and
PEX11 [9]. Drought stress also increases
transcription of the peroxisome biogene-
sis gene PEX11c in Arabidopsis [10].
Finally, the application of exogenous
H2O2 triggers the proliferation of peroxi-
somes ([8] and citations therein), demon-
strating a positive feedback loop between
Trends in Plant Science, Month Year, Vol. xx, No. yy 1
 TRPLSC 1615 No. of Pages 3

ROS accumulation and peroxisome

abundance. Thus, changes of ROS Reducon of soil moisture content
homeostasis in response to water deficit 1
could modulate peroxisome abundance
in cells, although direct experimental evi- ABA synthesis in roots
dence to support this interaction is still ABA moves to shoots 1
lacking. Below I propose a hypothetical
model for the impact of drought on per-
oxisome abundance. The production of
Stomata closure
ROS under other abiotic stress factors 2 2
such as salinity, heavy metals, tempera-
ture, etc. could also impact on peroxi- Chloroplast
Nucleus
some proliferation. The signal Light
transduction pathways in those cases DNA
would likely differ from [93_TD$IF]my model.
energy
ROS-scavengers 2
Peroxisome Abundance May and peroxisome 3
Inform Responses to Drought biogenesis genes 2 CO2
Stress 5 fixaon
Water deficiency is thought to be first
perceived by the root system (Figure 1). 4
Roots relay this signal to shoots through
the synthesis of abscisic acid (ABA),
5
5
ROS 3
3Lipids 4
6 3
which moves to leaves via the xylem Peroxisomes
sap. Upon reaching the leaves, ABA indu- proliferaon RNA Enzymes
ces closure of the stomata, thus reducing
water loss through transpiration. Stomata 6
closure limits the intake of CO2 and inhib- Synthesis of ROS
its carbon fixation. Consequently, residual
light energy that is not used in CO2 fixation
scavengers Cytoplasm
becomes converted into ROS through (i)
oxygen photoreduction by photosystems
I and II; (ii) the formation of singlet oxygen Figure 1. [90_TD$IF]Model of Crosstalk between Peroxisome Abundance and Reactive Oxygen Species
(1[91_TD$IF]O2), which results from transfer of light (ROS) Homeostasis under Drought. (1) Roots perceive reduced soil moisture content and produce
abscisic acid (ABA), which is transported to shoots. After reaching the shoots, ABA induces stomata closure.
energy from exited chlorophyll to ground- (2) Stomata closure reduces CO fixation and, as a consequence, more light energy collected by the
2
state oxygen; and (iii) an increase in chloroplast is converted to ROS. (3) ROS damage lipids, structural proteins, and enzymes, as well as
photorespiration. RNA and DNA, in all cellular compartments. (4) Damage to lipids and proteins further compromises CO2
fixation as well as other chemical reactions in the cell, and ROS production increases. (5) ROS activate the
expression of ROS scavengers and peroxisome biogenesis factors. (6) ROS neutralization alleviates oxidative
ROS can damage multiple cellular struc- damage to the cellular components. Consequently, the chances of cell survival during stress recovery would
tures through different mechanisms [11]: increase.
modification of guanine to form 8-hydrox-
yguanine by 1O2, which leads to muta- above types of damage inhibit photosyn- damage, and consequentially increase
tions in DNA and RNA; peroxidation of thesis, which in turn causes even higher the chances of plant survival following
polyunsaturated fatty acids by 1O2 and ROS production[94_TD$IF]. If it becomes too exten- drought.
hydroxyl radical, which causes a reduc- sive, the oxidative damage will further
tion in membrane fluidity and permeabil- compromise recovery from the stress Given this knowledge, my model pro-
ity, and perturbs the function of integral and may eventually lead to cell death. poses that
membrane proteins; and modification of Under conditions of severe water loss,
amino acids through nitrosylation, car- the only known intervention is the produc- [95_TD$IF](i) peroxisome abundance in cells and
bonylation, and/or oxidation, which tion of the ROS scavengers and the pro- organs can be used as a proxy of ROS
results in protein damage and inhibition liferation of peroxisomes, which may homeostasis: higher abundance would
of enzymes [11,12]. Collectively, the neutralize ROS, alleviate the oxidative be indicative of less efficient ROS

2 Trends in Plant Science, Month Year, Vol. xx, No. yy

 TRPLSC 1615 No. of Pages 3
homeostasis and vice versa. Measuring
ROS homeostasis to test this hypothesis
on a large scale, however, has been com-
plicated by the chemical diversity of ROS
and ROS scavengers, which requires
individualized techniques for the detec-
tion and quantification of each class of
molecules. For the same reason, high-
throughput assays for ROS have not
proved feasible. These challenges could
be overcome using a fluorescent probe
such as N-BODIPY to measure the abun-
dance of peroxisomes in a semi high-
throughput format [8], and
[96_TD$IF](ii) efficient ROS-scavenging systems in a
given genotype may correlate with
improved performance under drought.
Relatively fewer peroxisomes under
drought stress would be indicative of
reduced ROS content and better cellular
fitness. Higher peroxisome abundance
would represent ROS overproduction
and poor fitness.
Systematic analyses of the correlation
between peroxisome abundance and
yield traits [97_TD$IF]under drought stress would
demonstrate the agricultural usefulness
of this parameter. Considering the
variability of the drought-adaptation
strategies, some genotypes could
scavenge the excess of ROS through
a peroxisome-independent mechanism,
whereas other may avoid ROS
production. Further experimental work
would help to determine whether perox-
isomes play an active role in enhancing
drought tolerance (and can therefore be
manipulated for increased performance)
or are simply reporters that inform the
current state of ROS homeostasis in
the plant.
Acknowledgments
I appreciate the help of Mark Hubbard in writing
this manuscript. The laboratory of A.S. is supported
by the US Department of Agriculture (grant
WNPO3826), a Dr OA Vogel Foundation award, a
Washington State University (WSU) New Faculty Seed
Grant, the WSU BioAg fund, and a CRDF award (grant
62765).
1
Institute of Biological Chemistry, Washington State
University, Pullman, WA 99163, USA
*Correspondence:
andrei.smertenko@wsu.edu (A. Smertenko).
https://doi.org/10.1016/j.tplants.2017.09.021
References
1. Molden, D. et al. (2003) A water-productivity framework for
understanding and action. In Water Productivity in
Agriculture: Limits and Opportunities for Improvement
(Kijne, J.W., ed.), pp. 1–18, CABI
2. Salekdeh, G.H. et al. (2009) Conceptual framework for
drought phenotyping during molecular breeding. Trends
Plant Sci. 14, 488–496
3. Sinclair, T.R. (2011) Challenges in breeding for yield
increase for drought. Trends Plant Sci. 16, 289–293
4. Hu, J. et al. (2012) Plant peroxisomes: biogenesis and
function. Plant cell 24, 2279–2303
5. Foyer, C.H. and Noctor, G. (2003) Redox sensing and
signalling associated with reactive oxygen in chloroplasts,
peroxisomes and mitochondria. Physiol. Plant. 119,
355–364
6. Mittler, R. and Zilinskas, B.A. (1994) Regulation of
pea cytosolic ascorbate peroxidase and other antioxidant
enzymes during the progression of drought stress
and following recovery from drought. Plant J. 5,
397–405
7. Guo, Z. et al. (2006) Differential responses of antioxidative
system to chilling and drought in four rice cultivars differing
in sensitivity. Plant Physiol. Biochem. 44, 828–836
8. Fahy, D. et al. (2017) Impact of salt stress, cell death, and
autophagy on peroxisomes: quantitative and morphologi-
cal analyses using small fluorescent probe N-BODIPY. Sci.
Rep. 7, 39069
9. Charlton, W.L. et al. (2005) Salt-induced expression of
peroxisome-associated genes requires components of
the ethylene, jasmonate and abscisic acid signalling path-
ways. Plant Cell Environ. 28, 513–524
10. Li, J. and Hu, J. (2015) Using co-expression analysis and
stress-based screens to uncover Arabidopsis peroxisomal
proteins involved in drought response. PLoS One 10,
e0137762
11. Moller, I.M. et al. (2007) Oxidative modifications to cellular
components in plants. Annu. Rev. Plant Biol. 58,
459–481
12. Apel, K. and Hirt, H. (2004) Reactive oxygen species:
metabolism, oxidative stress, and signal transduction.
Annu. Rev. Plant Biol. 55, 373–399
Trends in Plant Science, Month Year, Vol. xx, No. yy 3