Growing rice without water:

Understanding rice drought

tolerance mechanisms
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Contents
1. Background .......................................................................................................................................... 1
2. Summary.............................................................................................................................................. 1
3. Introduction......................................................................................................................................... 2
3.1. Context analyses and justification of the intervention area in which the project takes place .... 2
3.2. Problem analysis........................................................................................................................... 2
4. Purpose / Results ................................................................................................................................. 3
5. Research methodology ........................................................................................................................ 3
General outline .................................................................................................................................... 3
Detailed description of tasks ............................................................................................................... 5
Task 1: Comparing wheat and three types of rice: Morphological and physiological traits (PhD1
India or China) ................................................................................................................................. 6
Task 2: Comparing wheat and three types of rice: Hormonal regulation and root architecture
(PhD2 WU) ....................................................................................................................................... 6
Task 3: Comparing wheat and three types of rice: System-level understanding of the whole-plant
physiology of acclimation and adaption to field stress (PhD3 IRRI or China or India) .................... 7
Task 4: Genome wide association mapping of drought related traits in rice (PhD4 China or IRRI) 8
Task 5: Transcriptomics, other molecular analyses, and systems integration (PhD5 WU) ............. 9
6. References ......................................................................................................................................... 10

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1. Background
Rice is grown under inundated conditions for over 5000 years, i.e. standing in a water layer of 5-10 cm.
Yet, the reasons for this practice are all agronomic as rice is not an exclusively aquatic species– meaning
that we should be able to change the practice for rice to be grown like wheat for instance. Inundation
helps to plough the heavy clay soils by oxen, it suppresses weeds while rice can stand the water, it
allows to store water from heavy rain showers during the monsoon season, it allows the fern Azolla to
grow on the water layer and fix nitrogen through symbiosis with a bacteria to fertilize the rice, it helps to
dissolve nutrients like phosphorus from the soil, and more. Increasingly rice is grown outside monsoon
seasons and in less heavy soils because of the increasing demand for rice. All practices can however be
altered e.g. by ploughing with tractors, by applying fertilizers, by weeding with tractors, spraying
herbicides, etc. Also, the availability of water is declining dramatically, the cultivation of rice requires
much labour, the wet conditions create much emission of greenhouse gasses, and other crops than rice
can be grown only with great difficulty following a rice crop because the soil is slammed. We have found
already that rice can be grown with half the amount of water but still requires muddy soils – yields
collapse once the soils start to crack. For this and other reasons farmers are reluctant to adopt the
current growing practices of rice with less water. A next step is to understand why rice cannot perform
like wheat when grown under comparable conditions as wheat. This would resolve most of the mentioned
problems with inundated rice cultivation and may allow gradual mechanisation of the system with
increasing efficiency of the use of scarce natural resources including water and nutrients.

2. Summary
Rice uses about 30% of the freshwater used for agricultural crops worldwide; exploring ways to
reduce water use for rice production is therefore of great strategic value for sustainable crop
production for the world facing water scarcity. Various water-saving management practices have
been designed to reduce water use by rice; however, yield penalty has generally been unavoidable.
There is a lack of information on characteristics of rice varieties that are required for superior
performance under water saving crop management. The main aim of this project is to unravel and
understand the morphological and physiological mechanisms determining the differences in water
requirement between rice and wheat. Two complementary approaches will be used to achieve this
aim. In Approach 1, wheat and three types of rice (lowland, aerobic and upland rice) are directly
compared for morphological and physiological characteristics. In Approach 2, genetical genomic
analysis using a rice population will be conducted. Using these two approaches, the whole project
will comprise five tasks, to be carried out by five PhD researchers that will intensively interact to
exchange insights, materials and data. It is expected that the mechanisms that lead to the
different behaviour of wheat and the various rice types and the underlying molecular regulation will
be unravelled. The mechanisms, markers and genes and their functions revealed in this project will
be of substantial value for breeders to select for new rice cultivars that have reduced water use at
a level similar to wheat without yield penalty.
3. Introduction

3.1. Context analyses and justification of the intervention area in which the
project takes place
Rice is the primary food for more than half of the world population, especially in developing
countries such as Asia, where water scarcity and drought are imminent threats to food security.
Rice uses two to five times more water than other cereal food crops such as wheat or maize and
uses about 30% of the freshwater used for agricultural crops worldwide. Exploring ways to reduce
water use for rice production is therefore of great strategic value for sustainable crop production
for the world facing water scarcity (Molden et al. 2010).

Lowland rice (including both irrigated and rain-fed lowland) contributes more than 90% of total rice
production. The irrigated rice crop is continuously grown in inundated fields (i.e. fields with a layer
of 5-10 cm water) and obtains high yields up to and over 10 tonnes per hectare but very low water
use efficiency (WUE; the amount of water used to produce 1 kg of dry matter). Traditional upland
rice varieties are adapted to growth in fields without a water layer but they have much lower yields
of about 2-4 tonnes per hectare. To reduce water use in inundated rice while maintaining its high
yield levels, water-saving regimes have been developed (Bindraban et al. 2006) including a system
of rice intensification SRI (Uphoff & Randriamiharison 2002), alternate wetting and drying irrigation
(AWD)(Bouman & Tuong 2001), controlled soil drying during grain filling (Yang & Zhang 2010) and
non-flooded mulching cultivation (Liu et al. 2005). Some of these practices can reduce water use
up to 50% without a yield penalty, but these practices are not adopted by farmers as they incur
too high risks. Further reducing water use in these strategies does improve WUE, but at the cost of
yield penalty (Yang & Zhang 2010). Attempts in China, India and at IRRI (International Rice
Research Institute, Philippines) to breed for so-called aerobic rice (growing under more dry
conditions, so not inundated or muddy) have resulted in varieties that use less water and have
reasonable yields of circa 6 tonnes per hectare. However, aerobic rice cannot replace lowland rice
in most of the rice growing areas due to its lower grain yield, and can be an option only for farmers
in rain-fed lowlands with limited or erratic rainfall (Atlin et al. 2006).

The need to further reduce water use which will allow cultivation of rice as a dry crop while
obtaining high yields remains essential. In addition to pursuing breeding strategies with upland and
lowland rice varieties, there is a need for a fundamental understanding of the physiological
functioning of rice. Why can rice not perform similar to other cereals in terms of its ability to take
up water and nutrients, retaining a high yield under non-inundated conditions?

Comparing upland rice with rice grown in paddies there are clear morphological differences: upland
varieties make longer roots and probably produce less shoot biomass, and this is likely one of the
main reasons why they can deal with dryer conditions (De Datta 1975). Other physiological
parameters that may affect water use efficiency are the number of stomata (or the stomatal
resistance), the root’s capacity to take up water and nutrients under dry conditions, the ability to
continue photosynthesis under dry conditions due to, for example, high hydraulic conductivity
(Adashi et al. 2011). According to Peng & Bouman (2007), there is little information on morpho-
physiological characteristics of varieties that are required for superior performance under water
saving crop management.

3.2. Problem analysis

Significant physiological differences are known to exist between crops. So called C4 crops such as
maize have, for instance, a higher photosynthetic rate, and therefore, a higher physiological WUE,
than C3 crops such as wheat. This is because the C4 photosynthetic pathway creates an elevated
internal CO2 concentration that largely prevents the occurrence of photorespiration (e.g. Yin et al.
2011), a process that reduces photosynthetic efficiency. Rice is a C3 crop like wheat and barley but
requires much more water than these other C3 crops to obtain high yields. While much of the water
is lost because of outflow of water from the field (i.e. seepage and percolation) and evaporation in
early growth stages, the sharp decline in yield under dryer conditions suggest specific features of

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rice. Rice indeed has different morph-physiological features as compared to wheat, such as the
ability to develop aerenchym to allow oxygen flow from the leaves to the roots that experience
anaerobic (oxygen-less) condition. The stomata of rice plants are smaller than in other plants
species but rice has a higher stomatal density (Sheehy et al. 2000). Despite these fundamental
differences in morphology and anatomy, the average physiological WUE (grain yield per unit of
water transpired) under their respective prevailing growth field environments was only slightly
lower in rice than in wheat and other C3 small grain cereals (Haefele et al. 2009).

To understand the different responses of wheat and rice to limited water availability, two things
merit attention. First, rice is commonly grown in warm environments, whereas wheat is generally
grown in temperate environments. It is well known that the difference in environmental conditions
such as in temperature and vapour pressure can cause a difference in WUE (Morison & Gifford
1983). Secondly, rice and wheat are different in the evolutionary patterns, through which adaptive
changes in morphology and physiology have occurred. Adapting to inundated conditions may have
led to the superficial rooting and shorter root hairs in rice, for instance. Therefore it is essential to
investigate the two crops and upland rice under the same growing conditions, to examine where
the crops differ and to find the reasons behind these differences.

4. Purpose / Results
Our general hypothesis is that through the evolutionary adaption to their growth environments,
wet and dry, respectively, rice and wheat have developed their specific strategies to cope with that
environment. These strategies work best only when their responses at all levels, including gene
expression, hormones levels, physiological and morphological adaptations are optimally tuned.
Therefore, we will examine differences in morphology and anatomy as well as in the physiological
mechanisms of responses to water availability between wheat and rice.

This project aims to unravel the mechanisms responsible for the differences in drought tolerance
and water use efficiency among various types of rice (upland, aerobic, lowland), as well as between
rice and wheat. We want to examine the following key questions:

(1) whether or not there is a continuum in the morphological and physiological

characteristics/mechanisms along the chain ‘wheat / upland rice / aerobic rice / lowland
rice’;
(2) which morphological and/or physiological characteristics are most important for the
evolutionary and adaptive patterns of rice types and wheat;
(3) What regulatory mechanisms govern those characteristics;
(4) what is the molecular-genetic basis of the differences in these morphological and
physiological traits

It is expected that these differences can ultimately be explored by agronomic and/or breeding
approaches to grow rice with reduced water use to the level of wheat without yield penalty.

5. Research methodology

General outline
As stated earlier, the main aim of this project is to unravel and understand the morpho-
physiological and molecular mechanisms determining the differences in water
requirement between rice and wheat. Two approaches will be used (Fig. 1). In approach 1,
wheat and three types of rice are compared. In this approach, inevitably choices have to be made
with respect to the processes/parameters to be studied, requiring a priori knowledge and

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hypotheses. However, it has the advantage of going beyond ‘rice’ and may reveal mechanisms not
(yet) known in rice. In Approach 2, genetical genomic analyses using a rice population will be
conducted. Approach 2 is unbiased, i.e. no a priori choice or hypotheses are made, and it may
reveal processes, not thought of before-hand. Thus the results from Approach 2 will direct further
studies in Approach 1, while findings in Approach 1 might be indicative for targeting studies in
Approach 2 (Fig. 1).

Fig. 1 The two approaches used in the project and the information flow between them.

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The two approaches are implemented, specifically focusing on:

(1) Comparing wheat and three different types of rice (upland, aerobic and lowland rice).
Plants will be grown at two or three different conditions (drought-stressed, well-watered,
and flooded) and a wide range of agronomic, physiological and morphological
characteristics will be determined (details below). This will indicate processes affected by
water availability and the differential responses by the species/varieties. 3 PhDs will
perform their research within the framework of this Approach 1, focusing on morphology
and anatomy, hormonal regulation and root architecture, and whole-plant physiology.
(2) Genetical genomic analyses of responses of rice to drought. Genetical genomics combines
the power of genomics (and other –‘omic’ approaches) with genetics, exploiting QTL
analysis and other ways to correlate molecular markers and traits. Besides being useful in
marker-assisted breeding (not the focus of this project) it has been shown to be powerful
in revealing mechanisms, and underpinning genes, involved in complex physiological
processes (e.g. Sergeeva et al. 2006). However, given that conventional QTL analysis of
rice drought tolerance has been extensively studied in the literature (e.g. Luza et al. 2010;
Gu et al. 2011; Venuprasad et al. 2011), this project will not conduct QTL mapping per se;
instead, a meta-analysis of the existing QTL studies combined with genome-wide
association mapping using a large number of rice accessions will be performed (1 PhD). In
addition, transcriptomics and other molecular analysis will be conducted (1 PhD), and this
PhD will use plant materials gathered by all other PhDs, thereby playing an integrative role.

Detailed description of tasks

With the two approaches described above, the whole project will comprise five tasks, to be carried
out by five PhD researchers that will intensively interact to exchange insights, materials and data
(Fig. 2). The team of supervisors will collectively govern the course of the research. The students
will study the mechanisms that lead to the different behaviour of wheat and the various rice types
and the underlying genetic regulation.

Fig. 2 Five PhD projects that will interact to exchange insights, materials and data, while each of them will use
approach 1 or 2 to implement a project task that has a specific research emphasis.

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The project will be carried out by 2 PhD students working in Wageningen University (WU), and 3
so-called sandwich PhD students in collaboration with three research institutes in the most
important rice growing countries: Philippines, China and India. Of them, three PhDs (1-3) will
largely follow Approach 1, one PhD (4) will follow Approach 2, and the last (PhD5) will, in principle,
play an integrative role although focusing on transcriptomics and other molecular analysis (Fig. 2).

Task 1: Comparing wheat and three types of rice: Morphological and physiological traits
(PhD1 India or China)
Main hypothesis
Rice has developed aerenchym to allow oxygen flow from the leaves, via stems, to the roots
that experience anaerobic condition. This system may be developed at the cost of reducing the
space within both root and stem tissues for the vascular transport system, xylem vessels in
particular. This hypothesis is supported by the fact that some aquatic plants are xylem-absent,
probably as result of or because of the need for the full aerenchym space. If the xylem vessels
or bundles are smaller in rice than in wheat, overall water transport will be slower since
transport rate depends on the overall cross-sectional area of the xylem bundle itself (Niklas
1985). For wheat, a breeding programme to select for narrow xylem vessels has resulted in 3-
11% yield improvement in dry environments because compared with unselected controls,
genotypes of reduced vessels can regulate crop water use so that more water is reserved in the
subsoil for later use after anthesis, resulting in higher harvest index and grain yield (Richards &
Passoura 1989). We hypothesise that the size of xylem vessels differs between rice and wheat,
and between different types of rice. It may also be possible that rice has different other
morphological characteristics for example, cuticle, stomata size and density, etc. We will
investigate their importance of these morphological and anatomical characteristics in
determining drought tolerance.

Materials and methods

Two lines of wheat and three different cultivation practices of rice (upland, aerobic and paddy)
will be grown under three conditions: limited water supply, well-watered and flooded (wheat will
not be grown under flooding) and a wide range of parameters will be determined, with an
emphasis on anatomy and morphology:
- shoot and root anatomy (aerenchym and xylem vessels)
- leaf morphology (stomatal density, distribution, length and width, and aperture)
- Morphology (cuticle structure and composition) and architecture (tillering, branching,
adventitious roots, lateral roots, root length, root hairs)
- Yield, yield components and crop WUE
- Stomatal conductance, photosynthesis, mesophyll conductance
- Carbon isotope discrimination (associated with leaf-level WUE) if possible
- collect a few overall growth characteristics (e.g. total biomass, plant height, grain yield) to
allow comparison with plants grown in task 2 and 3
- Drought stress marker gene expression in root and shoot (with PhD5)
- Take samples for transcriptomics and hormone analyses

Task 2: Comparing wheat and three types of rice: Hormonal regulation and root
architecture (PhD2 WU)
Main hypothesis
Growth and development of plants, and the responses of plants to environmental factors, are
largely controlled by plant growth regulators (hormones), e.g. abscisic acid (ABA) plays a
central role in coordinating responses to water loss (Chaves et al. 2003). ABA is produced both
in the roots as well as in the leaves and hence plays a role in local responses to drought as well

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 as in long distance signalling. Something similar holds for the new class of plant hormones, the
strigolactones. These compounds are principally produced in the roots and transported to the
shoot (Kohlen et al. 2011). They play a role in the regulation of shoot branching/tillering and
the shaping of root architecture. Strigolactone biosynthesis is strongly upregulated under
phosphate shortage as shown by us in several plant species, incl. rice (Jamil et al. 2011) and
the consequence is that plants reduce shoot branching/tillering and increase lateral root
formation. Strigolactones are also external signals that are secreted by plant roots into the
rhizosphere and are used by the symbiotic arbuscular mycorrhizal fungi for host detection. The
upregulation of strigolactone secretion under phosphate starvation is an adaptation to improve
the chances to initiate symbiosis with the AM fungi to improve phosphate uptake. Drought and
flooding may also affect nutrient availability and could therefore also affect strigolactone
formation. Other hormones that may play a role in the adaptation to flooding and drought are
ethylene (required for aerenchym formation but also involved in stomata regulation), and
cytokinin, brassinosteroids etc. (Peleg et al. 2011). We hypothesize that the interaction of
hormonal signalling cascades determines how plant organs respond to drought, which implies an
important role of hormones in the control of root growth during water stress.

Materials and methods

PhD2 will use the same varieties as PhD1 and grow them in the greenhouse in Wageningen for
detailed analyses of root architecture (using rhizotrons) and plant hormones:
- Grow upland, aerobic and paddy rice as well as wheat (2 varieties of each) under highly
controlled conditions, including three levels of water availability and collect root and shoot
samples for:
o Analysis of plant hormones known to be involved in root architectural changes and
drought response (ABA, auxin, cytokinin, strigolactones and their catabolites and
conjugates)
o gene expression profiling using RNAseq (with PhD5)
- collect a few overall growth characteristics (e.g. total biomass, plant height, grain yield) to
allow comparison with plants grown in task 1 and 3
- Use rhizotrons to grow the same varieties under the same conditions to analyse root
architecture (branching, adventitious roots, lateral roots, root length) in the different
varieties/species in response to drought.
- Study mycorrhizal colonisation under these conditions
- Use materials generated by PhD1 and PhD4 for plant hormone analysis (selection of lines
and conditions, bulk segregant analysis)
- Do experiments with hormone mutants and hormone applications to verify hypotheses on
plant hormone regulation

Task 3: Comparing wheat and three types of rice: System-level understanding of the
whole-plant physiology of acclimation and adaption to field stress (PhD3 IRRI or China or
India)
Main hypothesis
For this project, it will be ultimately needed to know the performance of crops under real field
environments, in which climatic variables and water availabilities are continuously changing. We
know that short-term and long-term responses of crops to water stress differ. In the short term
(< 12 h) plants responds by stomatal regulation and changes at the biochemical level. In the
mid-term (over days), various physiological processes acclimates to the growth environment by
changes in the plant proteome. Longer-term acclimation results in changes in plant structure.
Besides the short-term photosynthetic responses, we understand little of the system-level
mechanism of the whole-crop responses.
Long-term acclimation to stress, arising from the modification of morphological traits (e.g.
thicker leaves), have direct relevance on crop growth. Photosynthesis is hypothesised to play an
important role in this acclimation because the chloroplastic redox signalling as induced by an

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 imbalance between energy supply and demand acts with other pathways to elicit responses,
extending its influence beyond leaf mesophyll cells to influence meristem activity and organ
development resulting in changes to plant morphology. Investigation of the photosynthetic
acclimation mechanism based on energy balance and its after effects on morphology may
enable to better quantify season-long crop performance under fluctuating field environments,
from understandings of underlying basic processes. As affected by different morphology and
transport systems, wheat and rice, as well as different types of rice, are hypothesised to act in a
different pace for this acclimation, which in turn, has an impact on their intrinsic ability in coping
with water shortage.
For plants water does not only constitute around 90% of the (vegetative) parts, it is also
the vehicle for transport of nutrients, both in the soil, and in the plant. Thus shortage of water
may directly or indirectly affect nutrient availability and uptake. Therefore, in this task we will
also analyse if, and to what extent, water availability affects the nutrient status of plants by
analysing the most important ones (N, P, K, Ca, Fe).

Materials and methods

PhD3 will fully cooperate with PhD1 and PhD2, and try to translate the information on
morphology and hormonal regulation obtained in PhDs 1 and 2 into the understanding at the
whole-plant physiological level. In addition, (s)he will also use the same varieties as PhD1 and
PhD2 to grow them both in the greenhouse and in experimental fields to experimentally
characterise the plasticity of rice and wheat genotypes. Both lines of work will be conducted
from the perspective to understand plant acclimation strategies and crop productivity under field
conditions in response to short- to long-term water stress. Tasks of this PhD will include:
- Measurement of yield, yield components and WUE at crop (field) and plant (greenhouse)
levels;
- Measurement of hydraulic conductivity (Magnetic resonance imaging or other methods)
- Measurement of gas exchange and chlorophyll fluorescence (short-term response)
- in vitro analysis of chloroplast proteome (mid-term response)
- Measurement of leaf and plant morphology (long-term response)
- Measurement of stress recovery
- collect a few overall growth characteristics (e.g. total biomass, plant height, grain yield) to
allow comparison with plants grown in task 1 and 2
- analyse total nutrient (N, K, P, Fe, Ca) content
- Integration of these measurements and those from PhD1 and PhD2 into a whole-plant
physiology model to quantitatively assess the physiological requirements of hypothetical
rice that uses the same level of water as wheat does.

Task 4: Genome wide association mapping of drought related traits in rice (PhD4 China or
IRRI)
Background
Conventional QTL analysis of rice drought tolerance, often based on a bi-parental cross between
lowland and upland rice varieties, has been extensively studied in the literature. The intrinsic
drawback of QTL analysis using a bi-parental cross is the experimental setup in which only two
parents are involved; therefore, it is unlikely that the total genetic variation present in the
complete germplasm pool will be found. In addition, it is difficult to study many different traits
in a single test cross because it is difficult to find parents that are substantially contrasting for
all the traits under study. Association mapping, based on linkage disequilibrium (LD), in which
statistical association between genotypes and phenotypes is scrutinized over a large germplasm
collection, is now increasingly being recognized as a valuable addition to the toolbox for
identifying loci contributing to quantitative traits. The availability of whole-genome sequences in
some plant species like rice has released unprecedented information about polymorphism,
notably single nucleotide polymorphisms (SNP). High throughput SNP analysis allows whole-
genome molecular diversity characterization to localize favourable genes and alleles or

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 haplotypes through LD mapping. Comprehensive SNP data from a large collection of rice
accessions allows full dimension whole-genome LD scan in rice. This genome-wide association
(GWA) mapping, when compared with QTLs from multiple bi-parental populations, will reveal
unprecedented information for markers of drought tolerance. Co-localisation of QTLs/genes will
suggest mechanistic links between parameters (for example between plant hormones and root
architecture, between root architecture and drought tolerance, between stomatal conductance
and drought tolerance, etc). In order to optimize comparison between existing QTL data and the
results from our studies, we will try to include the parents of the existing bi-parental populations
in our studies. Transcriptome profiling of these lines, under contrasting conditions, in
combination with available QTL data will provide clues towards the understanding of underlying
mechanisms.

Materials & methods

Using wide diversity set of rice genotype accessions (available either from IRRI or from China?),
this PhD4 will perform the following experiments and analyses:
- Yield, a selection of root and shoot morphology parameters (cuticle structure and
composition, aerenchym) and architecture (shoot tillering, root branching, adventitious
roots, lateral roots, root length)
- Stomatal conductance, photosynthesis, mesophyll conductance
- Plant hormones
- GWA mapping will be used to map all the traits analysed (yield, WUE, morphological traits,
plant hormones, stomatal conductance, mesophyll conductance, gene expression, etc) to
the genome-wide SNP map of the population.
- Inventory study and meta-analysis of rice drought QTLs, and compare the results of the
meta-analysis with the GWA mapping.
- Do experiments to verify mechanisms and gene candidates together with PhDs 2 and 5.

Task 5: Transcriptomics, other molecular analyses, and systems integration (PhD5 WU)
Background
The capacity of a plant to adapt to various environments is determined by its genome, the blue
print of the genetic characteristics. Internal plant and external environmental triggers regulate
the differential expression of these characteristics, i.e. transcription from DNA to mRNA, and the
subsequent translation to proteins. These proteins function as enzymes or transporters, a major
set of proteins controls metabolic processes, including carbohydrate metabolism and transport
processes in plants, and with all this the ultimate growth and reproductive behaviour of the
plant. The rapidly increasing knowledge of genes and their function, allows classification of
genes into functional groups. Techniques are now available to make a profile of all transcripts
(mRNAs) of a plant or tissue simultaneously, and under various conditions, an approach
nicknamed ‘transcriptomics’. In combination with the functional classification of genes, it reveals
processes specifically up- or down-regulated under specific environmental conditions, and/or in
particular tissues. We will use this approach to analyse the samples of different lines, grown
under contrasting conditions, to trace relevant key-processes in rice and wheat.

Materials & methods

PhD 5 will be responsible for molecular analyses. He/she will carry out gene expression analysis,
bioinformatics, and systems biology:
- Transcriptomics of the materials generated by PhD1 and PhD2 for wheat and rice cultivars,
including the parents of existing mapping populations
- Bulk segregant samples from the GWA mapping population of PhD4: transcription profiling of
all lines of the diversity set is not feasible, but a selection of lines with interesting differences
will be analysed
- Do experiments, check gene expression, characterise mutants, make transgenic lines to
verify gene candidates together with other PhDs
- Development of Systems Biology approach (if possible) for integration at the molecular level

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