CSIRO PUBLISHING

Functional Plant Biology, 2012, 39, 851–859

http://dx.doi.org/10.1071/FP12079

Phenotyping for drought tolerance in grain crops:

when is it useful to breeders?

J. B. Passioura

CSIRO Plant Industry, PO Box 1600, Canberra, ACT 2601, Australia. Email: john.passioura@csiro.au

Abstract. Breeding for drought tolerance in grain crops is not a generic issue. Periods of drought vary in length, timing and
intensity and different traits are important with different types of drought. The search for generic drought tolerance using
single-gene transformations has been disappointing. It has typically concentrated on survival of plants suffering from severe
water stress, which is rarely an important trait in crops. More promising approaches that target complex traits tailored to
specific requirements at the different main stages of the life of a crop, during: establishment, vegetative development, floral
development and grain growth are outlined. The challenge is to devise inexpensive and effective ways of identifying
promising phenotypes with the aim of aligning them with genomic information to identify molecular markers useful to
breeders. Controlled environments offer the stability to search for attractive phenotypes or genotypes in a specific type of
drought. The recent availability of robots for measuring large number of plants means that large numbers of genotypes can be
readily phenotyped. However, controlled environments differ greatly from those in the field. Devising pot experiments that
cater for important yield-determining processes in the field is difficult, especially when water is limiting. Thus, breeders are
unlikely to take much notice of research in controlled environments unless the worth of specific traits has been demonstrated
in the field. An essential link in translating laboratory research to the field is the development of novel genotypes that
incorporate gene(s) expressing a promising trait into breeding lines that are adapted to target field environments. Only if the
novel genotypes perform well in the field are they likely to gain the interest of breeders. High throughput phenotyping will
play a pivotal role in this process.

Additional keywords: deficit watering, floral resilience, germplasm, prebreeding, trait, water stress.

Received 13 April 2012, accepted 5 September 2012, published online 25 September 2012

Introduction There are two, often overlapping uses for phenotypic

Last year (2011) was the centenary of the introduction by
Johannsen (1911) of the terms ‘genotype’ and ‘phenotype’
into the English language. He had earlier invented the terms in
his native Danish to further a debate with Francis Galton and
Karl Pearson, who had wrongly assumed that variation in a
given observed quantitative trait was entirely genetic and was,
therefore, of hereditary importance. Johannsen overturned their
view by demonstrating substantial variation in quantitative traits,
which he called ‘phenotypical’, in genetically-identical material.
The pheno- words soon became well established, though it
was not until about the 1950s that ‘phenotyping’ as a noun, ‘to
phenotype’ as a verb and ‘phenome’ as the collective noun, were
introduced. In the last 10–15 years their use has taken off
exponentially. The realisation had dawned that the richness of
the genomic information that had been accumulating
could not be well mobilised without connecting it to allied
phenotypic information. Johannsen’s insight into the diversity
of phenotypes arising from one genotype throws into relief the
difficulty of using phenotypic information to identify desirable
genotypes, for phenotypic variation within a genotype can
obscure phenotypic differences among genotypes. Pertinent
statistical analysis is needed.
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CSIRO 2012
information. One is to understand more deeply the functional
significance of particular genes or clusters of genes. The other is
to use it to develop selection tools in breeding, ideally robust
molecular markers, but also easily measured morphological,
developmental or physiological characteristics. Given the large
segregating populations used by plant breeders, there has been
much interest in rapidly phenotyping these large populations
(‘high throughput’). The question arises then of what traits would
be of most interest to plant breeders.
Breeders of crop plants are typically too busy to explore novel
traits that might or might not turn out to be useful. They are driven
by their principal requirement to produce cultivars that are
attractive to farmers. Several traits are essential: grain quality,
resistance to the most important diseases and developmental
alignment with the target environments. High yield is also
important, but it has such low heritability in drought-prone
environments that it can’t be selected for except in multisite
multiseason field trials of advanced breeding lines (Rebetzke et al.
2012). Even then, performance on farms may differ from those
of breeders’ trials, for farmers must deal with many management
constraints that breeders do not (Passioura 2010; Passioura and
Angus 2010). Nevertheless, Bänziger et al. (2006) have shown
www.publish.csiro.au/journals/fpb
852 Functional Plant Biology
how to increase heritability for water-limited yield in maize by
using carefully manipulated droughts.
Yet phenotyping for ability to deal effectively with abiotic
stress such as drought, waterlogging, salinity and temperatures
too high or too low, is an important pursuit in several
recently established plant phenotyping facilities http://www.
plantphenomics.com/partners/ (accessed 17 August 2012).
Much of this interest revolves around developing rapid
phenotyping techniques that aim to explore traits postulated to
improve resistance to a given stress or to discover new ones.
These traits are often explored initially in controlled laboratory
environments and may not be well connected to the way plants
behave season-long in the field (Passioura 2010). If they turn
out to not be well connected, there is little chance that breeders
will incorporate germplasm that expresses such traits into
their breeding programs. The practical rationale for the work is
thereby broken, though it does deepen genetic, biochemical and
physiological understanding, which may enable productive
avenues for improvement some decades hence.
Phenotyping for ‘drought tolerance’ or ‘drought resistance’
(these terms are now commonly used interchangeably), has
attracted much recent interest, with citation data increasing by
40% per year since 5 years ago when the interest started to
blossom. The purpose of this paper is to analyse various
approaches that have been and are being developed to breed
grain crops for water-limited environments with the help of
novel phenotyping, which can lead to the development of
novel germplasm expressing potentially useful traits that are
easy to select for and are adequately heritable. The challenge
is to create a continuous chain within this branch of agricultural
research that will eventually result in cultivars that are better
able to turn a limiting supply of water into grain yield. Most
rapid progress is likely to come from interactions across this
chain from the very beginning of a project. The role of breeders
involved in developing and field-testing novel germplasm is
pivotal.
Searching for drought-tolerant crops
‘Drought’ has many meanings, depending on one’s point of
view. Conversations about it are, in consequence, often at
cross purposes. Geographers, meteorologists, insurers,
breeders, agronomists, plant physiologists and molecular
biologists generally have their own operational meanings
(Passioura 2007). Similarly, within the plant sciences, ‘drought
tolerance’ (or ‘resistance’) has several meanings, ranging from
the ability to survive severe water deficits (e.g. resurrection
plants, cacti) to the ability of crops to use most effectively a
limited water supply – to capture more of the water that is
potentially available (essentially, the rain that falls while the
crop is growing plus the available water in the soil at the time of
sowing) and to make best use of that water in producing grain.
There are many aspects of the behaviour of crops, morphological,
physiological and biochemical, both above- and belowground,
which influence these components.
Breeders concerned with drought tolerance usually look for
lines that yield relatively well when water is scarce but do not lose
the ability to yield well in good seasons (Fleury et al. 2010).
Agronomists and crop physiologists tend to be more analytical.
J. B. Passioura
In the agronomic context, ‘drought’ can refer to unusually low
rainfall during the crop’s growing season (for example, the driest
10% of growing seasons), but it also refers to the timing of that
rainfall during the growing season, to episodes of dryness
that reduce yield. Water may be limiting at any stage of a
crop’s life: at sowing; during establishment and vegetative
growth; during sensitive times for maintaining floral fertility;
and during grain growth. There are no universal traits that cover
all of these possibilities. Agronomists deal with them and prepare
for them, individually.
The timing of flowering is the most important physiological
trait in water-limited environments. There is an optimal flowering
time that depends on the balance between water used during
canopy development and water used from around the time of
flowering to when the grain is ready to harvest. This optimum is
often tempered, in crops that grow during winter and spring, by
the need to flower late enough in the spring to avoid frosts during
flowering, which can lead to massive failure in producing grain.
At the same time it is important to (i) develop enough biomass
by the time of flowering to set up a good potential yield (the
potential number of grains m–2 is proportional to that biomass),
yet (ii) ensure that enough water remains available in the soil to
maintain the crop’s photosynthetic and translocation activity
during flowering and grain growth (Fischer 1979).
Each of these requirements can be dissected further
in contributing to the final yield (Passioura and Angus 2010).
They can be a source of ideas for selecting what traits to pursue,
what breeding lines to phenotype and what environments to tune
them to. In dealing with a large segregating population, what
traits are the ones that are most likely to be influential, have good
heritability and be easy to select by breeders? Traits that are not
easy to select for and whose worth (proof of concept) has not been
demonstrated in realistic environments, will typically be ignored
by breeders.
The following discussion deals with various approaches that
are now popular in searching for traits to improve the yields of
droughted crops in drought-prone environments. They can be
broadly classed into: (1) survival of severe water deficits, whether
of a whole plant or of its floral organs; and (2) productivity
issues that relate to the capture of available water and its most
effective use, essentially targeting what to do with water when it is
available, not when it is not. The latter also has two contrasting,
yet complementary, approaches: (i) an essentially observational
one, in which genotypic variation in the responses of plants to
modest changes in water supply is studied; and (ii) an engineering
one, in which genotypic variation in specific ‘designer’ traits is
explored.
Survival of severe water deficits
Ability of plants to survive severe water-deficits during
vegetative growth
Studying the genetics and performance of plants exposed to
severe water deficits has been widely used, especially in
molecular genetics. There are ~1000 journal papers in this
general area, 40% of which involve Arabidopsis. The work
typically involves transforming plants with genes whose
expression is postulated to protect plants or cellular functions
from severe water stress, though better survival of transformed
Phenotyping for drought tolerance
plants has often resulted from their being smaller and therefore,
using water more slowly than the wild types (Morran et al. 2011).
Although survival of severe water stress may be important for
the commercial production of turf grasses or perennial pasture
plants in arid or semiarid environments, it is irrelevant for
commercial grain crops; if a drought is so bad that it threatens
the survival of a crop then yields are certain to be disastrously
low whether the crop survives or not (Sinclair 2011). Farmers
are likely to harvest a dying crop for forage, which is usually
valuable in such circumstances. Nevertheless, this work has
produced major insights into the cellular basis of dehydration
tolerance, the role of the DREB/CBF family of transcription
factors being a notable example (Shinozaki and Yamaguchi-
Shinozaki 2000).
That said, one potentially useful trait is the ability of very
young seedlings to withstand severe water deficits. With the
expected increase in climatic variability, there is likely to be a
widespread trend in semiarid environments towards sowing at the
optimal time even if the soil is dry: http://www.csiro.au/en/
Outcomes/Food-and-Agriculture/Dry-sown-crops.aspx (accessed
17 August 2012). This has the advantage that sowing is done
without pressure of time and the seed is in position to respond
immediately to a moderate fall of rain. However, if the seeds
germinate and the rain is not followed within about a fortnight
by more rain, the seedlings could fail (Finch-Savage 2004; Finch-
Savage et al. 2010).
Maintenance of floral fertility despite water stress
Floral fertility can be markedly reduced by abiotic stress –
especially low water potential during pollen mother cell meiosis
and shortly before and after anthesis, which can affect the viability
of both pollen and ovules (Saini and Westgate 1999; Boyer and
McLaughlin 2007; Ji et al. 2010; Parish et al. 2012), as well as
leading to the mistiming of anthesis and silking in maize (Campos
et al. 2006). The resulting poor seed set leads to low yields despite
vegetative growth being good. This is an important topic. Crops
often behave too conservatively: low yield can result from
transient water stress at an inopportune time, stress that would
be of little concern in the vegetative phase of the crop. In maize,
Boyer and colleagues (summarised by Boyer and McLaughlin
2007) have shown that the processes taking place when maize
ovules abort during water stress are complex: they depend on the
duration of the stress and involve carbohydrate metabolism and
the induction of senescence genes. Lesions in carbohydrate
metabolism are also involved when water stress results in the
failure of pollen to develop (Dolferus et al. 2011); genotypes of
wheat with good survival of pollen have been found.
Physiological and morphological behaviour of plants
in drought-prone environments
Genotypic variation in responses to changing water supply
The recent development of robots that can measure the
growth and in some cases water consumption of large numbers
of plants at frequent intervals has stimulated interest in diversity
in the responses of crop plants to changes in their water supply.
In general, genotypes that keep growing fast in the face of a
decreasing water supply perform well in the field whenever
there is adequate water in the subsoil and the prospects of rain
Functional Plant Biology 853
or irrigation are good, so that there is productive advantage in the
crop continuing to grow fast (Tardieu 2012); rice is a good
example (Parent et al. 2010).
In many circumstances, especially in semiarid environments,
conservative behaviour during the main period of vegetative
growth is often beneficial. Slow growth and thus slow water
use in response to a developing soil water deficit conserves water
in the subsoil for use later in the season during the critical periods
of flowering and grain growth. This too is a different sort of
‘drought tolerance’ – the marshalling of a limiting water supply
to achieve the most productive seasonal distribution of water
use. It is notable that dual-purpose wheat crops (used for both
grazing and producing grain) that are grazed to ground level
before stem elongation sometimes yield substantially more grain
than ungrazed controls when water-limited, seemingly because of
deferred water use (Harrison et al. 2011).
Targeting traits for optimising water balance to achieve
water-limited potential yield
Different traits assume importance at different stages of
crop development. Many of these relate to making appropriate
use of water when it is available, often with the aim of
ensuring that adequate water is available during the sensitive
times of floral development, flowering and grain growth. They
include morphological traits, such as root architecture
(Hammer et al. 2009; Lopes and Reynolds 2010; Trachsel
et al. 2011) and reduction in unproductive tillers in cereals
(Mitchell et al. 2012) and physiological ones, such as the
retranslocation of pre-anthesis assimilate to the grain (Bidinger
et al. 1977; Blum 1998). A trade-off between root depth and
amount of stored assimilate may be a concern (Lopes and
Reynolds 2010), although Kirkegaard et al. (2007) showed
that the uptake of only an extra 10 mm of water by wheat roots
from the deep subsoil during grain growth led to an extra
600 kg ha–1 of grain yield, presumably largely because of more
effective transfer of stored assimilate to the grain.
Reynolds and Tuberosa (2008) and Richards et al. (2010) have
discussed a wide range of traits for improving water-limited yield
in wheat and Salekdeh et al. (2009) have done so for a wide range
of crops. In summary, these include: good seedling establishment
(e.g. long coleoptiles); rapid ground cover; leaf architecture; root
vigour; transpiration efficiency (carbon-isotope discrimination);
remobilisation to the grain of stem carbohydrates; glaucousness
to deflect heat; leaf rolling; and buffering against reproductive
failure. Maintenance of green leaf area (stay-green) and cool
canopies during grain filling also look promising (Jordan et al.
2012; Lopes and Reynolds 2012). Vigorous growth of seedlings
is strongly beneficial in unploughed soil, a characteristic of
conservation agriculture (Watt et al. 2005). Deep roots in rice
are evidently beneficial (Henry et al. 2011) and in maize better
buffering of floret fertility and early grain filling against water
stress are likely to be the next areas of improvement after
the earlier success in reducing the stress-induced gap between
anthesis and silking (ASI) (Campos et al. 2006). Further,
promising new traits are sure to emerge as work in this area
continues to blossom.
It is notable that some of these traits are constitutive and can be
selected for in well-watered plants (Richards et al. 2010):
854 Functional Plant Biology
(a) long coleoptiles to enable better establishment when seeds
are sown deeply to access water below a dry surface soil
(coleoptiles of the semi-dwarf wheats that contain the
dwarfing genes Rht1 and Rht2 are unable to elongate more
than ~50 mm, so seeds sown more deeply than this produce
seedlings that fail to emerge);
(b) fast development of early leaf area to make use of water that
might otherwise evaporate from a wet soil surface; and
(c) reduced tillering to avoid wasteful investment in
unproductive tillers.
Given these diverse approaches for improving drought
tolerance or water-limited yield in annual crops, how can we
make good progress using controlled environments, which offer
much better control for pursuing promising traits, but which are
limited in their ability to reproduce the field conditions that most
influence the final grain yield?
Phenotyping in controlled environments aimed
at crop improvement
Many important yield-determining processes in the field take
place slowly; for example, the slow extraction of water from
subsoil that often takes place over several weeks. It is difficult to
mimic these using pot experiments in controlled environments.
Passioura (2006), Poorter et al. (2012a, 2012b) and Salekdeh
et al. (2009) have discussed a wide range of issues in running pot
experiments.
Particular difficulties with experiments dealing with water
stress are that pots are usually much smaller than the volume of
soil available to a plant in the field (Poorter et al. 2012a) and the
softness of the growing medium enables much faster exploration
by roots and faster leaf growth than usually happens in the
field (Stirzaker et al. 1996; Passioura 2002). The rate of use of
the available water supply can, therefore, be many times faster
than in the field, leading to exhaustion over days rather than
weeks or even months. One result is that fast responses by the
plants, for example, stomatal closure, which can be effective over
minutes to hours, tend to overshadow slow but often ultimately
more influential processes such as the development of root growth
into previously untapped wet parts of the soil and the modulation
of leaf area index to adjust to a slowly depleting store of soil water
(Jordan 1983).
Generally, integrative measurements are more useful than
spot measurements in elucidating the main influences on
water-limited yield (Masuka et al. 2012), although persistent
patterns of stomatal behaviour, for example, midday stomatal
closure, may be beneficial in modulating water use (Rebetzke
et al. 2003).
Nevertheless, there are some one-off fast events in the field that
can strongly influence grain yield. Frost at the time of flowering is
one and sudden hot dry wind at the time of flowering or during
grain growth is another. The risk of frost at flowering interacts
with the risk of severe water deficits during grain growth of cereal
crops growing during winter and spring (http://www.grdc.com.
au/uploads/documents/GRDC_FS_TimeOfSowing_North.pdf?
shortcut=1, accessed 17 August 2012) and is, therefore, pertinent
to improving the water management by such crops. These fast
events may be easier to explore in controlled environments than in
the field, although it has proven difficult to use frost chambers to
J. B. Passioura
replicate the relative behaviour of different genotypes
experiencing damaging frost in the field (Chen et al. 2009;
Frederiks et al. 2011).
What measurements to make
There is a wide range of physiological and morphological
measurements that can be made on plants growing in pots:
water potential, stomatal conductance, leaf temperature,
various characteristics of photosynthesis, multispectral
reflectance, thermal imaging, biomass, rate and amount of
water use, relative growth rate and leaf area development, root
growth and distribution and many others. Some of these can be
done effectively and non-destructively with remote sensing
(Furbank and Tester 2011; Masuka et al. 2012; Nagel et al.
2012). If the eventual aim is to incorporate promising traits into
commercial breeding lines, choosing measurements that enable
rapid selection of plants in populations segregating for such
traits is likely to make the fastest progress. Often, promising
traits are first identified in the field, for example, the need for long
coleoptiles in wheat, to enable better establishment of seedlings
when there is a dry start to a season (Rebetzke et al. 2007). Others
have arisen from laboratory or theoretical studies, such as the use
of carbon isotope discrimination to select for intrinsic water-use
efficiency (Condon et al. 2004), that is, the ratio of biomass
produced to the amount of water transpired.
The appropriate measurements to make are those that either
directly concern the given trait or that are a good surrogate
for it. For example, cool canopies during grain filling, easily
measurable by thermal imagery, may be a useful surrogate for
root depth (Lopes and Reynolds 2010). Similarly, the diurnal
pattern of stomatal conductance, also rapidly measurable by
thermal imagery, may be an indicator of parsimonious and
efficient water use during the vegetative phase (Rebetzke et al.
2003). Phenomics, although in its infancy, is developing new
measurement techniques, sometimes with no specific traits in
mind. Some of these techniques are sure to prove valuable, but
they are likely to be most rapidly effective if, during their
development, they are related to traits of known or potential
importance.
How best to simulate in controlled environments a drought
experience or modulation of water use that gets close to what
happens in the field?
There are many aspects of pot experiments that involve the water
(and oxygen) relations of crop plants and can strongly influence
behaviour. At best, these experiments are based on hypotheses
about what traits are likely to be influential in improving yield in a
range of circumstances in the field, or on what new procedures can
speed up the rate of selection of traits that may be important, but
are currently too cumbersome to be useful in breeding programs.
There are many traits, outlined earlier, that are currently of
interest, many of which are amenable to selection in controlled
environments and some of which are best selected for in well
watered conditions.
Various watering regimes have been tried for reducing the
water supply to plants growing in pots with the aim of looking
for useful genetic variation. Each has its advantages and
disadvantages in relation to the specific aims of the research.
Phenotyping for drought tolerance Functional Plant Biology 855

In some studies, good technique requires that the surface of the
medium be covered so that there is little loss of water by direct
evaporation from the soil. Doing so enables the amount of water
transpired by the plants to be measured, which in turn, can be
related to the performance of the plant in relation to its water use. If
this is not done, a slowly growing genotype will lose a larger
proportion of its water supply to direct evaporation, yet might
have a larger ratio of biomass accumulation to water transpired.
In these circumstances it would be difficult to interpret any
variation of the plants’ behaviour in relation to its water economy.
Tables 1 and 2 summarise some features of pot experiments
and of the water relations of the growth medium that can
influence interpretations of how the plants behave. They
compare common techniques for establishing well-watered
controls and for applying deficit watering.
Prebreeding for effective use of phenomic information – the
link to breeding programs
Prebreeding, formerly known as genetic enhancement (Duvick
1990), is succinctly described by FAO’s Global Partnership
Initiative for Plant Breeding Capacity Building (GIPB) as:
‘. . . all activities designed to identify desirable characteristics
and/or genes from unadapted materials that cannot be used
directly in breeding populations and to transfer these traits to
an intermediate set of materials that breeders can use further in
producing new varieties for farmers. It is a necessary first step in
the use of diversity arising from wild relatives and other
unimproved materials.’ http://www.bioversityinternational.org/
fileadmin/bioversityDocs/Training/Pre-breeding_manual/PB-A4-
Presentation_en.pdf (accessed 17 August 2012).
Implicit in this description is that the worth of the modified
breeding populations needs to be well demonstrated in the field if
they are to attract the attention of commercial breeders. Richards
et al. (2010) describe how near-isogenic or recombinant inbred
lines (RILs) can be developed in which given traits are either
present or absent in a common background. These lines can then
be grown in the field, preferably with managed water deficits, so
that the worth of a given trait can be assessed in different types of
drought (Campos et al. 2004; Rebetzke et al. 2012).
Commercial breeders of annual crops are unlikely to take
much interest in evidence from controlled environments that a
given trait may be beneficial. Given that different traits assume
greater or lesser importance in different types and timing of
drought, they are much more likely to take interest in a trait
that has passed these additional tests:
(i) climatic analysis has shown that it is likely to be important in
a substantial proportion of growing seasons over a large
target area (e.g. Chenu et al. 2011);
(ii) the relevant genes have been incorporated into cultivars or
advanced breeding lines that are adapted to the target areas
(Rebetzke et al. 2012);
(iii) lines contrasting in expression of the trait (high and low)
have been grown in plots in the field and the presence of
the trait has evidently improved the water-limited yield
of the plants;
(iv) the trait is not deleterious in seasons without drought or
is not excessively prone to trade-offs in different types of
drought (Tardieu 2012);
(v) the trait is genetically stable, persists across generations and
is expressed in different genetic backgrounds; and
(vi) the trait is easy to select for in breeding populations, either
with robust molecular markers, rapid visual appraisal, or
automated remote sensing.
Traits that concern the performance of plants at different
developmental stages are likely to be effective in only a
minority of growing seasons, e.g. one in four. For example, if
conditions at the time of sowing are good, then traits that
help establishment in poor conditions are not relevant.
Similarly deep roots may not be of much importance during
droughts in which only the top 50 cm of the soil profile gets water
and similarly in good seasons, with ample well distributed
rainfall, when water deep in the subsoil, though present, is not
needed (Lilley and Kirkegaard 2007).
There is an important corollary to this. In commercial breeding
programs, selection is strongly driven by the essentials of
disease resistance, grain quality, right flowering time and,
finally, in the generation or two before the release of a new
cultivar, yield (although obviously underperforming lines are
culled in all generations). This means that specific traits that have
been deliberately incorporated into breeding lines are often not
explicitly selected for in later generations. Thus, if there is a
majority of growing seasons in which a given trait is of little
value, it may slowly disappear by attrition during the selection
process. A historical example is that of osmotic adjustment, which

Table 1. Comparison of experimental techniques for maintaining well-watered controls in pots

Technique Requirements Comment

Maintain ‘pot capacity’ (i.e. water frequently to
A
Highly porous artificial potting mix with many The suction immediately after draining ranges
temporary excess and allow to drain) pores >3 mm diameter to ensure adequate from 0 (i.e. saturated) at the base of the pot, to
aeration throughout the pot after watering 1 kPa per 100 mm of pot height (e.g. 2 kPa at the
top of a 200 mm tall pot)
Maintain ‘field capacity’A (which is the almost- Pots must be watered to the appropriate weight Typically corresponds to a soil water suction of
stable water content that develops in a recently- consistent with the water content at ‘field ~20 kPa (equivalent to that at the top of a freshly
watered freely-draining soil) capacity’ drained 2 m tall pot).Can use field soil with little
risk of hypoxia
A
These terms are discussed more fully in Passioura (2006) and in PrometheusWiki: http://prometheuswiki.publish.csiro.au/tiki-ndex.php?page=Soil
+Waterandhighlight=soil%20water (accessed 17 August 2012). An important feature of field capacity is that a clear wetting front develops as water
redistributes into a dry layer.
856 Functional Plant Biology J. B. Passioura

Table 2. Comparison of various techniques for deficit watering of plants in pots

Technique Purpose Comments

Stop watering until most plants look dead, then
rewater
Terminal drought (i.e. no further water added)
Brief period without watering, during which
the plants clearly respond to the lack of
water, but without experiencing severe
water deficits (vegetative plants)
As above, but targeting floral development
Rewatering by weight to a lower soil water
content than that of the control
Constant low rate of watering (below that
of the control)
Search for plants that can survive severe water
deficits
Search for plants that can make the most of
the existing water in the pots
Search for plants that either (a) grow as fast as
the well watered controls or (b) slow growth
substantially (i.e. respond conservatively)
Search for plants that can maintain floral
fertility despite water stress at sensitive
stages
Explore response of plants to water deficits
of various degrees
Explore response of plants to water deficits
of various degrees
May be useful for selecting more robust perennial pasture
plants, but failure to survive is as yet of little interest
to breeders of annual crops. Survival of very young
seedlings sown dry may become more important (see
earlier)
Ability to simulate environment in which crops experience
a terminal drought, e.g. throughout their lives, as with
winter/spring crops sown into a wet soil in a summer
rainfall environment, or during grain filling in some
Mediterranean environments
Plants that maintain fast growth are desirable where there is
little chance of soil water running out. Conservative
plants may be useful where the water supply is limited;
conserving water for later in the plant’s life, during
flowering and grain-filling, is often beneficial. In the
field, drying of the topsoil may be accompanied by
reduced availability of nutrients; exploring that
interaction in pots set up with nutrient supply decreasing
with depth may be of interest
An important trait with great potential for improving yields
under drought
Hard to relate to season-long performance of field-grown
plants. A complication is that fast-growing plants that
use more water between waterings will receive more
water (i.e. conservative plants may be inadvertently
disadvantaged). Uneven distribution of water in pot
may lead to chemical signalling associated with partial
root zone drying (Davies et al. 2002) because watering
to substantially less than field capacity creates wetting
fronts (see Table 1)
Avoids the possible problem that conservative plants may
be disadvantaged, but uneven distribution of water
could still be influential. However, if the water is
provided at the bottom of a long pot, this technique can
simulate the slow uptake of water from the subsoil that is
common during grain-filling in many drought-prone
environments

can be of substantial value in water-limited wheat (Morgan and
Condon 1986), yet there is no evidence that its frequency in
breeding populations has increased. Hammer et al. (2005) and
Messina et al. (2011) have provided frameworks for guiding
breeding goals where there are known interactions of this sort
between genotype and target environment.
Prospects for good progress
High-throughput phenotyping for improving performance under
drought is in its infancy. A growing array of new tools are
available for both understanding more deeply the functional
significance of particular identified genes and for selecting
desirable quantitative traits in prebreeding and breeding
programs (Furbank and Tester 2011; Masuka et al. 2012;
Nagel et al. 2012; White et al. 2012) both in the laboratory
and in the field. How does it stand in relation to other techniques
for facilitating breeding aimed at generating improved cultivars
for water-limited environments? Such techniques include genetic
transformation and the ‘black-box’ statistical technique of
genomic selection (Goddard 2009; Cabrera-Bosquet et al. 2012).
The technique of single-gene transformation has been very
successful with simple constitutive traits like resistance to pests
or herbicides that deal with alien molecules uninvolved in the
processes within the plant that contribute to yield (Sinclair 2011).
Resistance to pests or diseases is doubly valuable in that it can
enable more effective selection for higher yield by removing
sources of error that can obscure otherwise useful variation. With
abiotic stresses genetic transformation is at its best in unraveling
membrane and cellular functions and has led, for example, to
substantial progress in improving tolerance to aluminium (Pereira
et al. 2010), after astute molecular and physiological research
had identified and cloned important genes. Prospects are also
good for substantially improving durable resistance to fungal
pathogens with the recent cloning of rust-resistance genes in
wheat (Risk et al. 2012).
However, with complex multi-faceted traits such as drought
tolerance, contributions from genetic transformation have been
Phenotyping for drought tolerance
slight. The general reasons why are outlined by Araus et al.
(2008), Passioura (2010) and Sinclair (2011). Of the ~1000
papers that have been published that mention ‘drought
tolerance (or resistance) and transformation and gene’ in their
abstracts very few report evaluation in field plots. Two recent and
notable examples are Castiglioni et al. (2008) and Saint Pierre
et al. (2012). The first of these claimed some improvement of
yield when maize transformed with a bacterial RNA chaperone
experienced drought at flowering; however, little agronomic
information was supplied with which to gauge the nature of
the improvement and its likelihood of being usefully robust
across a range of environments – for example, was flowering
time precisely the same across the transformants and wild type?
The second showed that wheat plants transformed with DREB
did not yield more when grown with limiting water in the field,
although their ratio of shoot biomass to water used was greater in
pot experiments.
The main reasons for this general failure of genetic
transformation in relation to drought tolerance in the field is
that, first, most of the initial selection and identification of
candidate genes has relied on survival of extreme water
deficits and, as noted earlier in this paper, survival of severe
water deficits is not agronomically important, except possibly
for very young seedlings; and, second, it is rare for single genes
to contribute much to improvements in quantitative traits: the
recent success of genomic selection highlights that, for its power
has been in selecting for many quantitative trait loci (QTL) of
small but additive effect (Goddard 2009).
Genomic selection is proving to have great ability in predicting
breeding value within existing breeding populations. It requires
densely-mapped genotypes accompanied by rich phenotypic
information of a reference population. If the markers are dense
enough to be closely associated with useful QTL, they can be used
to select for favourable alleles at each QTL without needing to
identify the QTL or its functional significance (Goddard 2009).
It has the potential, despite the low heritability of yield in
drought-prone environments, of identifying the best genotypes
and perhaps of aligning them with different target environments
(Burgueño et al. 2012). Its efficacy may be further improved
by using overt knowledge of the roles of known QTL in
dealing with genotype-environment interactions (Messina
et al. 2011). Nevertheless, making progress in strongly
drought-prone environments in which yields can vary several-
fold across years will be difficult (Richards et al. 2010), especially
given the agronomic flexibility – particularly in sowing time and
fertiliser application – that farmers show in dealing with drought
(Passioura and Angus 2010). High-throughput phenotyping in the
field using remote sensing of traits that are associated with water-
limited yield (White et al. 2012) may help structure a reference
population for genomic selection.
Genomic selection is limited in its power to the reference
population on which it is trained. Thus, the pursuit of new traits
may require deliberate broadening of a breeding population.
A classic example of such broadening is the use of Norin 10 to
develop semi-dwarf wheats in the 1950s and ’60s (Hall and
Richards 2012) and a more recent example is the search
for alternative dwarfing genes that do not overly restrict the
length of wheat coleoptiles as the Norin 10 genes do (Ellis
et al. 2005).
Functional Plant Biology 857
It is in the exploration of novel traits that phenotyping in
controlled environments offers fastest initial progress in
identifying genotypes from which proof of concept in water-
limited field conditions can eventually by explored. Successful
examples, mentioned earlier, include transpiration efficiency,
coleoptile length and resilience (or not, depending on the
target environment) of leaf growth to water deficits.
Where will new ideas for traits effective in water-limited
environments come from? Observations of field scientists –
breeders, agronomist, crop physiologists – have been a fertile
source and will surely remain so. These can be thought of as
top down (in relation to the rest of the biological spectrum) and
come with the advantage that they are demonstrably relevant
in the field; long coleoptiles and the association between yield
and cool canopies during grain filling come into this class. Such
observations stimulate more detailed work in laboratory and
controlled environments, which in turn often results in greater
insights into limiting processes in the field. At the same time,
tuning suites of traits to target environments will be needed
(Chenu et al. 2011).
Meanwhile, phenomics is going through a phase of rapid
development in which new techniques are being developed
to explore morphological and physiological traits in a range of
genotypes, by direct robotic measurement, remote sensing or
both. In the context of breeding better cultivars of grain crops for
drought-prone environments, the challenge ahead is to make best
use of these techniques to explore, perhaps uncover, novel traits
and associated genotypes that can contribute to faster genetic
improvement. Cross fertilisation between laboratory and field is
needed (Salekdeh et al. 2009).
Given that the rates of genetic improvement required in the
coming decades are greater than those currently being achieved
(Fischer and Edmeades 2010) and that the lead times for trait-
based breeding have typically been ~20 years (Hall and Richards
2012), well-focussed high-throughput phenotyping will be
crucially important in developing novel effective traits. The
latter will require developing enhanced germplasm in adapted
genetic backgrounds that can be used in the field to obtain proof-
of-concept for such traits. Further development will then depend
on commercial breeders being sufficiently impressed with the
novel germplasm to use it in their breeding programs.
Acknowledgements
I thank Rana Munns, Hendrik Poorter, Greg Rebetzke and Richard Richards
for their valuable comments on this manuscript.
References
Araus JL, Slafer GA, Royo C, Dolores Serret M (2008) Breeding for yield
potential and stress adaptation in cereals. Critical Reviews in Plant
Sciences 27, 377–412. doi:10.1080/07352680802467736
Bänziger M, Setimela PS, Hodson D, Vivek B (2006) Breeding for improved
abiotic stress tolerance in maize adapted to southern Africa. Agricultural
Water Management 80, 212–224. doi:10.1016/j.agwat.2005.07.014
Bidinger F, Musgrave RB, Fischer RA (1977) Contribution of stored pre-
anthesis assimilate to grain yield in wheat and barley. Nature 270,
431–433. doi:10.1038/270431a0
Blum A (1998) Improving wheat grain filling under stress by stem reserve
mobilisation. Euphytica 100, 77–83. doi:10.1023/A:1018303922482
858 Functional Plant Biology
Boyer JS, McLaughlin JE (2007) Functional reversion to identify
controlling genes in multigenic responses: analysis of floral abortion.
Journal of Experimental Botany 58, 267–277. doi:10.1093/jxb/erl177
Burgueño J, de los Campos G, Weigel K, Crossa J (2012) Genomic prediction
of breeding values when modeling genotype  environment interaction
using pedigree and dense molecular markers. Crop Science 52, 707–719.
doi:10.2135/cropsci2011.06.0299
Cabrera-Bosquet L, Crossa J, von Zitzewitz J, Serret MD, Araus JL (2012)
High-throughput phenotyping and genomic selection: the frontiers of
crop breeding converge. Journal of Integrative Plant Biology 54,
312–320. doi:10.1111/j.1744-7909.2012.01116.x
Campos H, Cooper A, Habben JE, Edmeades GO, Schussler JR (2004)
Improving drought tolerance in maize: a view from industry. Field
Crops Research 90, 19–34. doi:10.1016/j.fcr.2004.07.003
Campos H, Cooper M, Edmeades GO, Loffler C, Schussler JR, Ibanez M
(2006) Changes in drought tolerance in maize associated with fifty
years of breeding for yield in the US corn belt. Maydica 51, 369–381.
Castiglioni P, Warner D, Bensen RJ, Anstrom DC, Harrison J, Stoecker
M, Abad M, Kumar G, Salvador S, D’Ordine R, Navarro S, Back
S, Fernandes M, Targolli J, Dasgupta S, Bonin C, Luethy MH,
Heard JE (2008) Bacterial RNA chaperones confer abiotic stress
tolerance in plants and improved grain yield in maize under water-
limited conditions. Plant Physiology 147, 446–455. doi:10.1104/
pp.108.118828
Chen A, Gusta LV, Brule-Babel A, Leach R, Baumann U, Fincher GB, Collins
NC (2009) Varietal and chromosome 2H locus-specific frost tolerance
in reproductive tissues of barley (Hordeum vulgare L.) detected using a
frost simulation chamber. Theoretical and Applied Genetics 119,
685–694. doi:10.1007/s00122-009-1079-1
Chenu K, Cooper M, Hammer GL, Mathews KL, Dreccer MF, Chapman SC
(2011) Environment characterization as an aid to wheat improvement:
interpreting genotype-environment interactions by modelling water-
deficit patterns in north-eastern Australia. Journal of Experimental
Botany 62, 1743–1755. doi:10.1093/jxb/erq459
Condon AG, Richards RA, Rebetzke GJ, Farquhar GD (2004) Breeding
for high water-use efficiency. Journal of Experimental Botany 55,
2447–2460. doi:10.1093/jxb/erh277
Davies WJ, Wilkinson S, Loveys B (2002) Stomatal control by chemical
signalling and the exploitation of this mechanism to increase water use
efficiency in agriculture. New Phytologist 153, 449–460. doi:10.1046/
j.0028-646X.2001.00345.x
Dolferus R, Ji X, Richards RA (2011) Abiotic stress and control of grain
number in cereals. Plant Science 181, 331–341. doi:10.1016/j.plantsci.
2011.05.015
Duvick DN (1990) Genetic enhancement and plant breeding. In ‘Advances in
new crops’. (Eds J Janick, JE Simon) pp. 90–96. (Timber Press: Portland,
OR)
Ellis MH, Rebetzke GJ, Azanza F, Richards RA, Spielmeyer W (2005)
Molecular mapping of gibberellin-responsive dwarfing genes in bread
wheat. Theoretical and Applied Genetics 111, 423–430. doi:10.1007/
s00122-005-2008-6
Finch-Savage WE (2004) The use of population-based threshold models to
describe and predict the effects of seedbed environment on germination
and seedling emergence of crops. In ‘Handbook of seed physiology.
Applications to agriculture’. (Eds R Benech-Arnold, RA Sanchez)
pp. 51–95. (Food Products Press: New York)
Finch-Savage WE, Clay HA, Lynn JR, Morris K (2010) Towards a genetic
understanding of seed vigour in small-seeded crops using natural variation
in Brassica oleracea. Plant Science 179, 582–589. doi:10.1016/j.plantsci.
2010.06.005
Fischer RA (1979) Growth and water limitation to dryland wheat yield in
Australia: a physiological framework. Journal of the Australian Institute
of Agricultural Science 45, 83–94.
Fischer RA, Edmeades GO (2010) Breeding and cereal yield progress. Crop
Science 50, S85–S98. doi:10.2135/cropsci2009.10.0564
J. B. Passioura
Fleury D, Jefferies S, Kuchel H, Langridge P (2010) Genetic and genomic
tools to improve drought tolerance in wheat. Journal of Experimental
Botany 61, 3211–3222. doi:10.1093/jxb/erq152
Frederiks TM, Christopher JT, Fletcher SEH, Borrell AK (2011) Post
head-emergence frost resistance of barley genotypes in the northern
grain region of Australia. Crop and Pasture Science 62, 736–745.
doi:10.1071/CP11079
Furbank RT, Tester M (2011) Phenomics – technologies to relieve the
phenotyping bottleneck. Trends in Plant Science 16, 635–644.
doi:10.1016/j.tplants.2011.09.005
Goddard M (2009) Genomic selection: prediction of accuracy and
maximisation of long term response. Genetica 136, 245–257.
doi:10.1007/s10709-008-9308-0
Hall AJ, Richards RA (2012) Prognosis for genetic improvement of yield
potential and water-limited yield of major grain crops. Field Crops
Research. doi:10.1016/j.fcr.2012.05.014
Hammer GL, Chapman S, Van Oosterom E, Podlich DW (2005) Trait
physiology and crop modelling as a framework to link phenotypic
complexity to underlying genetic systems. Australian Journal of
Agricultural Research 56, 947–960. doi:10.1071/AR05157
Hammer GL, Dong Z, McLean G, Doherty A, Messina C, Schusler J,
Zinselmeier C, Paszkiewicz S, Cooper M (2009) Can changes in
canopy and/or root system architecture explain historical maize yield
trends in the US corn belt? Crop Science 49, 299–312. doi:10.2135/
cropsci2008.03.0152
Harrison MT, Evans JR, Dove H, Moore AD (2011) Dual-purpose cereals:
can the relative influences of management and environment on crop
recovery and grain yield be dissected? Crop and Pasture Science 62,
930–946. doi:10.1071/CP11066
Henry A, Gowda VRP, Torres RO, McNally KL, Serraj R (2011) Variation in
root system architecture and drought response in rice (Oryza sativa):
Phenotyping of the OryzaSNP panel in rainfed lowland fields. Field Crops
Research 120, 205–214. doi:10.1016/j.fcr.2010.10.003
Ji X, Shiran B, Wan J, Lewis DC, Jenkins CLD, Condon AG, Richards RA,
Dolferus R (2010) Importance of pre-anthesis anther sink strength for
maintenance of grain number during reproductive stage water stress in
wheat. Plant, Cell & Environment 33, 926–942. doi:10.1111/j.1365-
3040.2010.02130.x
Johannsen W (1911) The genotype conception of heredity. American
Naturalist 45, 129–159. doi:10.1086/279202
Jordan WR (1983) Whole plant response to water deficits: an overview. In
‘Limitations to efficient water use in crop production’. (Eds HM Taylor,
WR Jordan, TR Sinclair) pp. 289–317. (American Society of Agronomy:
Madison, WI)
Jordan DR, Hunt CH, Cruickshank AW, Borrell AK, Henzell RG (2012)
The relationship between the stay-green trait and grain yield in elite
sorghum hybrids grown in a range of environments. Crop Science 52,
1153–1161. doi:10.2135/cropsci2011.06.0326
Kirkegaard JA, Lilley JM, Howe GN, Graham JM (2007) Impact of subsoil
water use on wheat yield. Australian Journal of Agricultural Research
58, 303–315. doi:10.1071/AR06285
Lilley JM, Kirkegaard JA (2007) Seasonal variation in the value of subsoil
water to wheat: simulation studies in southern New South Wales.
Australian Journal of Agricultural Research 58, 1115–1128.
doi:10.1071/AR07046
Lopes MS, Reynolds MP (2010) Partitioning of assimilates to deeper roots is
associated with cooler canopies and increased yield under drought in
wheat. Functional Plant Biology 37, 147–156. doi:10.1071/FP09121
Lopes MS, Reynolds MP (2012) Stay-green in spring wheat can be determined
by spectral reflectance measurements (normalized difference vegetation
index) independently from phenology. Journal of Experimental Botany
63, 3789–3798. doi:10.1093/jxb/ers071
Masuka B, Araus JL, Das B, Sonder K, Cairns JE (2012) Phenotyping for
abiotic stress tolerance in maize. Journal of Integrative Plant Biology 54,
238–249. doi:10.1111/j.1744-7909.2012.01118.x
Phenotyping for drought tolerance
Messina CD, Podlich D, Dong ZS, Samples M, Cooper M (2011) Yield-trait
performance landscapes: from theory to application in breeding maize
for drought tolerance. Journal of Experimental Botany 62, 855–868.
doi:10.1093/jxb/erq329
Mitchell JH, Chapman SC, Rebetzke GJ, Bonnett DG, Fukai S (2012)
Evaluation of a reduced-tillering (tin) gene in wheat lines grown across
different production environments. Crop and Pasture Science 63,
128–141. doi:10.1071/CP11260
Morgan JM, Condon AG (1986) Water-use, grain-yield, and osmoregulation
in wheat. Australian Journal of Plant Physiology 13, 523–532.
doi:10.1071/PP9860523
Morran S, Eini O, Pyvovarenko T, Parent B, Singh R, Ismagul A, Eliby S,
Shirley N, Langridge P, Lopato S (2011) Improvement of stress tolerance
of wheat and barley by modulation of expression of DREB/CBF factors.
Plant Biotechnology Journal 9, 230–249. doi:10.1111/j.1467-7652.2010.
00547.x
Nagel K, Putz A, Gilmer F, Heinz K, Fischbach A, Pfeifer J, Faget M,
Blossfeld S, Ernst M, Dimaki C, Kastenholz B, Kleinert A, Galinski A,
Scharr H, Fiorani F, Schurr U (2012) GROWSCREEN-Rhizo is a novel
phenotyping robot enabling simultaneous measurements of root and
shoot growth for plants grown in soil-filled rhizotrons. Functional
Plant Biology 39, 891–904. doi:10.1071/FP12023
Parent B, Suard B, Serraj R, Tardieu F (2010) Rice leaf growth and water
potential are resilient to evaporative demand and soil water deficit once
the effects of root system are neutralized. Plant, Cell & Environment 33,
1256–1267.
Parish RW, Phan HA, Iacuone S, Li SF (2012) Tapetal development and
abiotic stress: a centre of vulnerability. Functional Plant Biology 39,
553–559. doi:10.1071/FP12090
Passioura JB (2002) Soil conditions and plant growth. Plant, Cell &
Environment 25, 311–318. doi:10.1046/j.0016-8025.2001.00802.x
Passioura JB (2006) The perils of pot experiments. Functional Plant Biology
33, 1075–1079. doi:10.1071/FP06223
Passioura J (2007) The drought environment: physical, biological and
agricultural perspectives. Journal of Experimental Botany 58, 113–117.
doi:10.1093/jxb/erl212
Passioura JB (2010) Scaling up: the essence of effective agricultural research.
Functional Plant Biology 37, 585–591. doi:10.1071/FP10106
Passioura JB, Angus JF (2010) Improving productivity of crops in water-
limited environments. Advances in Agronomy 106, 37–75. doi:10.1016/
S0065-2113(10)06002-5
Pereira JF, Zhou GF, Delhaize E, Richardson T, Zhou MX, Ryan PR (2010)
Engineering greater aluminium resistance in wheat by over-expressing
TaALMT1. Annals of Botany 106, 205–214. doi:10.1093/aob/mcq058
Poorter H, Bühler J, van Dusschoten D, Climent J, Postma JA (2012a) Pot size
matters: a meta-analysis of the effects of rooting volume on plant growth.
Functional Plant Biology 39, 839–850. doi:10.1071/FP12049
Poorter P, Fiorani F, Stitt M, Schurr U, Finck A, Gibon Y, Usadel B, Munns R,
Atkin OK, Tardieu F, Pons TL (2012b) The art of growing plants
for experimental purposes: a practical guide for the plant biologist.
Functional Plant Biology 39, 821–838. doi:10.1071/FP12028
Rebetzke GJ, Condon AG, Richards RA, Farquhar GD (2003) Gene action
for leaf conductance in three wheat crosses. Australian Journal of
Agricultural Research 54, 381–387. doi:10.1071/AR02151
www.publish.csiro.au/journals/fpb
Functional Plant Biology 859
Rebetzke GJ, Richards RA, Fettell NA, Long M, Condon AG, Forrester RI,
Botwright TL (2007) Genotypic increases in coleoptile length improves
stand establishment, vigour and grain yield of deep-sown wheat. Field
Crops Research 100, 10–23. doi:10.1016/j.fcr.2006.05.001
Rebetzke GJ, Barrett-Lennard EG, Bennett D, Biddulph B, Chenu K, Deery
DM, Mayer J, Moeller C, Rattey AR (2012) A multi-site, managed
environment facility (MEF) for targeted trait and germplasm
phenotyping. Functional Plant Biology 40, in press.
Reynolds M, Tuberosa R (2008) Translational research impacting on crop
productivity in drought-prone environments. Current Opinion in Plant
Biology 11, 171–179. doi:10.1016/j.pbi.2008.02.005
Richards RA, Rebetzke GJ, Watt M, Condon AG, Spielmeyer W, Dolferus R
(2010) Breeding for improved water productivity in temperate
cereals: phenotyping, quantitative trait loci, markers and the selection
environment. Functional Plant Biology 37, 85–97. doi:10.1071/FP09219
Risk JM, Selter LL, Krattinger SG, Viccars LA, Richardson TM, Buesing G,
Herren G, Lagudah ES, Keller B (2012) Functional variability of the Lr34
durable resistance gene in transgenic wheat. Plant Biotechnology Journal
10, 477–487. doi:10.1111/j.1467-7652.2012.00683.x
Saini HS, Westgate ME (1999) Reproductive development in grain crops
during drought. Advances in Agronomy 68, 59–96. doi:10.1016/S0065-
2113(08)60843-3
Saint Pierre C, Crossa JL, Bonnett D, Yamaguchi-Shinozaki K, Reynolds MP
(2012) Phenotyping transgenic wheat for drought resistance. Journal of
Experimental Botany 63, 1799–1808. doi:10.1093/jxb/err385
Salekdeh GH, Reynolds M, Bennett J, Boyer J (2009) Conceptual framework
for drought phenotyping during molecular breeding. Trends in Plant
Science 14, 488–496. doi:10.1016/j.tplants.2009.07.007
Shinozaki K, Yamaguchi-Shinozaki K (2000) Molecular responses to
dehydration and low temperature: differences and cross-talk between
two stress signaling pathways. Current Opinion in Plant Biology 3,
217–223.
Sinclair TR (2011) Challenges in breeding for yield increase for drought.
Trends in Plant Science 16, 289–293. doi:10.1016/j.tplants.2011.02.008
Stirzaker RJ, Passioura JB, Wilms Y (1996) Soil structure and plant growth:
impact of bulk density and biopores. Plant and Soil 185, 151–162.
doi:10.1007/BF02257571
Tardieu F (2012) Any trait or trait-related allele can confer drought tolerance:
just design the right drought scenario. Journal of Experimental Botany
63, 25–31. doi:10.1093/jxb/err269
Trachsel S, Kaeppler SM, Brown KM, Lynch JP (2011) Shovelomics: high
throughput phenotyping of maize (Zea mays L.) root architecture in the
field. Plant and Soil 341, 75–87. doi:10.1007/s11104-010-0623-8
Watt M, Kirkegaard JA, Rebetzke GJ (2005) A wheat genotype developed for
rapid leaf growth copes well with the physical and biological constraints
of unploughed soil. Functional Plant Biology 32, 695–706. doi:10.1071/
FP05026
White JW, Andrade-Sanchez P, Gore MA, Bronson KF, Coffelt TA, Conley
MM, Feldmann KA, French AN, Heun JT, Hunsaker DJ, Jenks MA,
Kimball BA, Roth RL, Strand RJ, Thorp KR, Wall GW, Wang G (2012)
Field-based phenomics for plant genetics research. Field Crops Research
133, 101–112. doi:10.1016/j.fcr.2012.04.003