Inducing DT in Plants Review

Biotechnology Advances 28 (2010) 169183
Contents lists available at ScienceDirect
Biotechnology Advances
j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / b i o t e c h a d v
Research review paper
Inducing drought tolerance in plants: Recent advances

M. Ashraf
Department of Botany, University of Agriculture, Faisalabad, Pakistan
a r t i c l e
i n f o
Article history:
Received 21 September 2009
Received in revised form 6 November 2009
Accepted 9 November 2009
Available online 13 November 2009
Keywords:
Drought tolerance
Conventional breeding
Marker assisted selection
Quantitative trait loci
Transgenic plants
Osmolytes
Hormones
Recombinant inbred lines
a b s t r a c t
Undoubtedly, drought is one of the prime abiotic stresses in the world. Crop yield losses due to drought
stress are considerable. Although a variety of approaches have been used to alleviate the problem of drought,
plant breeding, either conventional breeding or genetic engineering, seems to be an efcient and economic
means of tailoring crops to enable them to grow successfully in drought-prone environments. During the last
century, although plant breeders have made ample progress through conventional breeding in developing
drought tolerant lines/cultivars of some selected crops, the approach is, in fact, highly time-consuming and
labor- and cost-intensive. Alternatively, marker-assisted breeding (MAB) is a more efcient approach, which
identies the usefulness of thousands of genomic regions of a crop under stress conditions, which was, in
reality, previously not possible. Quantitative trait loci (QTL) for drought tolerance have been identied for a
variety of traits in different crops. With the development of comprehensive molecular linkage maps, markerassisted selection procedures have led to pyramiding desirable traits to achieve improvements in crop
drought tolerance. However, the accuracy and preciseness in QTL identication are problematic.
Furthermore, signicant genetic environment interaction, large number of genes encoding yield, and use
of wrong mapping populations, have all harmed programs involved in mapping of QTL for high growth and
yield under water limited conditions. Under such circumstances, a transgenic approach to the problem
seems more convincing and practicable, and it is being pursued vigorously to improve qualitative and
quantitative traits including tolerance to biotic and abiotic stresses in different crops. Rapid advance in
knowledge on genomics and proteomics will certainly be benecial to ne-tune the molecular breeding and
transformation approaches so as to achieve a signicant progress in crop improvement in future. Knowledge
of gene regulation and signal transduction to generate drought tolerant crop cultivars/lines has been
discussed in the present review. In addition, the advantages and disadvantages as well as future prospects of
each breeding approach have also been discussed.
2009 Elsevier Inc. All rights reserved.
Contents
1.
2.
3.
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Conventional breeding for drought tolerance . . . . . . . . . . . . . . .
Marker-assisted breeding (MAB) for drought tolerance . . . . . . . . . .
3.1.
Identication of QTL associated with drought tolerance . . . . . . .
3.2.
Manipulation of QTL for developing drought tolerant crop cultivars/lines
4.
Engineering crops for enhanced drought toleranceTransgenic approach . . .
5.
Conclusions and future challenges . . . . . . . . . . . . . . . . . . . .
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1. Introduction
Plant breeding, conventional breeding or genetic engineering, is an
art through which crop varieties of high quality and yield are developed.
E-mail address: ashrafbot@yahoo.com.

0734-9750/$ see front matter 2009 Elsevier Inc. All rights reserved.
doi:10.1016/j.biotechadv.2009.11.005
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
169
170
171
171
173
176
180
181
Breeding for any desired trait undoubtedly requires a signicant amount

of genetic variation at intra-specic, inter-specic or inter-generic levels.
Variability, however, can be achieved by new gene combinations,
intercrossing those genotypes that hold desirable characteristics and
introducing new germplasm from other existing breeding programs
(Ashraf, 1994; Flowers, 2004; Ashraf and Akram, 2009). The variability
achieved through these means can be narrowed down by selecting a few
170
M. Ashraf / Biotechnology Advances 28 (2010) 169183
genotypes that excel in the target environment (Bnziger et al., 2004).

Considerable improvement in a trait can be made, if the genetic variance
among the genotypes of a crop, selection intensity, and heritability are
reasonably high (Falconer, 1989).
Plant breeding has contributed to a large extent in tackling the challenges of food security at global level. The contributions of plant breeding
to food production at global level have been enormous during the 20th
century. There has been most important plant breeding break-through for
almost all commercially important crops including major ones such as
maize, wheat, rice, cotton etc. The Green Revolution, which started in the
1940s and mainly based on traditional breeding, resulted in a phenomenal
increase in wheat and rice yield in many parts of the world and especially
in South Asia (Rajaram, 2005). Dr. Norman Borlaug (Founder of the Green
Revolution) and his team spent almost two decades breeding high
yielding dwarf wheat that was able to resist plant pests and diseases. The
dwarf wheat out-yielded the traditional varieties about two to three times.
However, relatively little breeding work has been carried out on
improving crops for drought tolerance. The achievements made so far in
improving drought tolerance of different crops through the integration of
conventional breeding, marker-assisted breeding (MAB) and genetic
engineering (transgenic approach) have been discussed in the present
review. MAB and transgenic approach are diverse biotechnologies, because through the earlier, desirable genes can be tagged so they can be
easily selected within the breeding population, whereas through the
latter, desirable genes can be transferred from one species to another. A
large number of genomic regions of a crop germplasm can be examined
for their breeding value through MAB, which facilitates the breeder to pool
genes of diverse origins (Vinh and Paterson, 2005; Humphreys and
Humphreys, 2005). In fact, this was not possible before through classical
breeding. In contrast, through the transgenic approach specic cloned
genes can be incorporated into an organism by limiting the transfer of
undesirable genes from the donor organism. Furthermore, pyramiding of
genes with similar effects is possible through this approach (Ashraf et al.,
2008; Gosal et al., 2009). However, both MAB and transgenic approaches
are deemed efcient and precise ways of improving a desired trait. They
are being used widely these days to generate stress tolerant cultivars/lines
of different crops. Recent progress made in exploiting the knowledge of
gene regulation and the phenomena involved therein in developing
drought tolerant crop cultivars/lines has also been discussed in the
present review.
2. Conventional breeding for drought tolerance
Through conventional breeding, genetic variability for drought
tolerance among crops/crop cultivars or among sexually compatible
plant species can be identied, and the genetic variation so identied can
be introduced through different mating designs into cultivars/lines with
good agronomic characteristics (Pocket). During the last century,
conventional breeders at different renowned international research
centers have made considerable strides in developing drought tolerant
lines/cultivars of some important food crops. For example, breeding
approach started at the International Maize and Wheat Improvement
Center (CIMMYT), Mexico in the 1970s for developing drought tolerant
maize is worth mentioning. Based on the selection and breeding program,
maize yield improvement of 59 to 233 kg ha 1 cycle 1 of recurrent fullsib or S1 selection, was recorded (Bnziger et al., 2004). The CIMMYT
breeding program was very outcome-oriented and multi-faceted, because
it focused on a multitude of imperative problems including drought,
low N, and common leaf and ear diseases (Bnziger et al., 2004). In 1997,
CIMMYT spanned its breeding program to southern Africa aimed at
improving maize for the drought-hit areas. A number of maize hybrids
developed by the CIMMYT scientists were found superior to all those
developed by private enterprises, in terms of growth and grain yield under
drought-prone environments (Bnziger et al., 2004). Plant breeders, at the
Crops Research Institute (CRI) based at Kumasi, Ghana, have developed a
highly drought tolerant maize cultivar Obatanpa GH in 2006 in
collaboration with the International Institute of Tropical Agriculture

(IITA), Ibadan, the CIMMYT, Mexico, and the Sasakawa Global 2000
(Published online April 25, 2006). Similarly, 16 early maturing maize
inbred lines (from TZEI 1 to TZEI 16) resistant to a scrounging weed Striga
hermonthica (Del.) Benth. were produced by the IITA. All these lines were
found to be highly resistant to water limited conditions (Published online
April 25, 2006).
Wheat, which is one of the important staple food crops of the world,
is adversely affected by drought. In view of a projection by Rajaram
(2001) more than 50% of the 237 million ha area in the world under
wheat cultivation is affected by periodic drought. As stated earlier, improvement in drought tolerance of a crop through selection and breeding
requires a substantial magnitude of heritable variation. If variation in the
existing germplasm of a crop is low then wild relatives may serve as a
rich source of appropriate genetic variation. At CIMMYT, a new synthetic
hexaploid has been developed by crossing the diploid wild ancestor,
Aegilops tauschii (goat grass) with tetraploid durum wheat (Triticum
turgidum var. durum). These hexaploid synthetics containing a complete
D-genome from A. tauschii have provided a signicant new variation for
tolerance to both biotic and abiotic stresses (Villareal et al., 1994;
Valkoun, 2001). At CIMMYT, more than 1000 accessions of A. tauschii
have been evaluated and new hexaploid lines developed. A signicant
new genetic variation in these newly developed hexaploid wheat has
been observed for abiotic stresses including drought stress (Valkoun,
2001). Useful variation for drought tolerance has also been identied in
Triticum urartu, T. boeticum, T. dicoccoides (Valkoun, 2001) and Aegilops
geniculata (Zaharieva et al., 2001). However, in view of Skovmand et al.
(2001) A. taushii is the predominant source of variation for drought
tolerance.
The International Center for Agricultural Research in Dry Areas
(ICARDA), Aleppo, Syria, and the International Crops Research Institute
for the Semi-Arid Tropics (ICRISAT), Andhra Pradesh, India, have a
similar research mandate, i.e. to improve the major staple food crops of
the dry regions particularly falling in Asia, the Middle East and Africa.
Plant breeders at these two sister institutes have focused mainly on the
premier dryland cereals such as barley, millet, sorghum and groundnut,
and leguminous pulse crops such as lentil, chickpea, pigeonpea, and faba
beans (World's Dryland Farmers New Agricultural Technology-Green
Revolution Never Reached Them.mht). Although efforts are underway at
both centers to develop drought tolerant varieties of different crops
mentioned earlier, the plant breeders at ICARDA have recently developed a new variety of barley which they claim as the world's most
drought tolerant barley variety (ICARDA News). In fact, the empirical
breeding strategies were employed to develop the variety by crossing a
land race with a wild barley line from Palestine. It has been reported that
the new drought-hardy barley variety produced 50% more grain yield
than that produced by ordinary barley lines/cultivars under dryland
conditions.
The International Rice Research Institute (IRRI) based at Las Baos,
Philippines, though is focusing primarily on quality and yield improvement in rice, efforts are also currently underway to develop
drought tolerant rice keeping in view of the fact that over 50% of the
world's rice is cultivated in rain-fed areas where the crop experiences
intermittent drought (MacLean et al., 2002).
A number of drought resistant cultivars/lines of different crops
registered so far in Crop Science or reported in other sources have been
listed in Table 1. These cultivars, undoubtedly, have been developed
solely using different protocols/designs of the conventional breeding
approach. These drought tolerant lines of different crops provide a
sound testament that conventional plant breeding played a considerable role during the last century not only for improving the quality and
yield of crops, but also for improving abiotic stress tolerance including
drought tolerance. However, now there is a general consensus of the
plant breeders that empirical plant breeding is a highly time-consuming
as well as a cost- and labor-intensive approach. While transferring
desired genes from one plant to other through the conventional plant
breeding, a number of undesired genes are also transferred. Furthermore, to achieve a desired gain through traditional breeding, a number
of selection and breeding cycles may be required. However, improvement in a trait through conventional breeding is not possible if the
appropriate genetic variation in the gene pool of a crop is either very low
or absent. The limited success in improving crop drought tolerance
could be due to the reason that the drought tolerance trait is controlled
by multiple genes having additive effect and a strong interaction exists
between the genes for drought tolerance and those involved in yield
potential. Thus, there is a need to seek more efcient approaches for
genetically tailoring crops for enhanced drought tolerance.
3. Marker-assisted breeding (MAB) for drought tolerance
Through marker-assisted breeding (MAB) it is now possible to
examine the usefulness of thousands of genomic regions of a crop
germplasm under water limited regimes, which was, in fact,
previously not possible. By examining the breeding value of each of
the genomic regions, the breeder can coalesce genes of multifarious
origins in novel ways, which was not possible previously with
conventional breeding tools and protocols (Concept Note).
3.1. Identication of QTL associated with drought tolerance
Like tolerance to other abiotic stresses, that to drought stress is
controlled by many minor genes (polygenes) that have additive
effects in their expression (Zhao, 2002; Mohammadi et al., 2005; Thi
Lang and Chi Buu, 2008). Thus, the loci on chromosomes housing such
types of genes are now referred to as quantitative trait loci (QTL).
Natural genetic variation of a crop can be exploited either via direct
selection under stressful conditions whether simulated or natural or
via mapping of QTL (polygenes) and subsequent marker-assisted
selection (Ashraf et al., 2008). QTL mapping allows to assess the
locations, numbers, magnitude of phenotypic effects, and pattern of
gene action (Vinh and Paterson, 2005). The role of polygenes in
controlling a trait has been widely assessed by traditional means, but
the use of DNA markers and QTL mapping has made it convenient to
dissect the complex traits (Humphreys and Humphreys, 2005). For a
QTL analysis, phenotypic evaluation is carried out of a large number of
plants from a population segregating for a variety of genetic markers;
then a part or the whole population is genotyped; and nally
appropriate statistical analysis is performed to pinpoint the loci
controlling a trait (Asins, 2002). Due to the intricacy of abiotic stress
tolerance and the problems encountered in phenotypic based
selection, the QTL mapping has been considered as imperative to
the use of DNA markers for improving stress tolerance (Ashraf et al.,
2008). Ashraf et al. (2008) have listed a variety of DNA markers such
as RFLPs, RAPDs, CAPS, PCRindels, AFLPs, microsatellites (SSRs), SNPs,
and DNA sequences being currently in use to examine the inheritance
of stress tolerance. QTL mapping for the drought tolerance trait has
been done in different crops, the most notable being maize, wheat,
barley, cotton, sorghum, and rice (Quarrie et al., 1994; Teulat et al.,
1997; Sari-Gorla et al., 1999; Saranga et al., 2001; Sanchez et al., 2002;
Bernier et al., 2008).
In cotton, using F3 families derived from the cross, Gossypium
barbadense cv. F-177 and Gossypium hirsutum cv. Siv'on, Saranga et al.
(2001) identied a subset of 33 QTL under water limited regime, i.e., 11
QTL for plant productivity, 5 for some key physiological traits, and 17 for
ber quality. Recently, using marker-assisted selection, near-isogenic
lines were produced through exchanging QTL for yield and some
drought-related traits between G. barbadense cv. F-177 and G. hirsutum
cv. Siv'on (Levi et al., 2009a,b). For most of the traits studied, the nearisogenic lines showed a marked adaptation to drought, but not for yield.
In particular, the G. barbadense near-isogenic lines showed a steady
photosynthetic efciency under varying water limited regimes.
171
In barley, QTL involved in some key growth and water relation

attributes were detected using 187 recombinant inbred lines (RILs)
resulting from a cross between two Mediterranean cultivars, Tadmor and
Fr/Apm (Teulat et al., 1997). QTL involved in variation in relative water
content (RWC), number of tillers (NL) and total shoot fresh mass were
found to be located on a RFLP-RAPD genetic map. It was also found that
different DNA regions mediate in constitutive water stress responses.
Under water limited regime, although one region on chromosome 1 was
found to be mainly involved in variation for RWC and NL, other map
locations were also found for RWC and NL. It is imperative to note that
epistatic interactions among many QTL and between QTL and other
markers were observed only under drought conditions, which suggest
that some chromosomal regions are denitely involved in controlling the
expression of the traits under water limited conditions.
In sorghum (Sorghum bicolor), several linkage maps have been
generated using RFLP and other known DNA markers (Xu et al., 1994;
Taramino et al., 1997; Rami et al., 1998; Kong et al., 2000; Sanchez et al.,
2002). In view of a report (Bowers et al., 2003), over 2400 loci have been
mapped on an F2 population developed from an inter-specic cross, S.
bicolor S. propinquum. For ne mapping of genes and QTL, the mean
marker density of 0.5 cM or 350 kb between DNA markers is appropriate
(Sanchez et al., 2002). However, using recombinant inbred lines (RILs)
and near-isogenic lines (NILs), Sanchez et al. (2002) detected several
genomic regions linked to resistance to pre-owering and post-owering drought stress. They found four distinct genomic regions involved in
the stay-green trait using a RIL population derived from the cross,
B35 Tx7000. These four stay-green QTL expressed repeatedly in all eld
trials they conducted and explained 53.5% of the phenotypic variance.
QTL studies for the stay-green trait proved useful in the identication of a
number of genomic regions associated with drought resistance.
In maize, a linkage analysis between the manifestation of some key
characters like male and female owering time, anthesis-silking
interval, plant height, and molecular markers [RFLP, microsatellites
(SSR) and AFLP] was carried out under different water regimes using a
maize population consisting of 142 RILs derived from selng the F1
population from a cross B73 H99 (Sari-Gorla et al., 1999). Linkage
analysis showed that, the QTL identied for male owering time and
plant height were the same under well-watered and water-stressed
conditions. In contrast, for female owering time and anthesis-silking
interval, the expression of QTL was different under normal conditions
or under drought stress. Feng-ling et al. (2008) developed a maize
segregating population from the cross N87-1 (drought resistant) 9526 (drought-sensitive), which was genotyped at 103 SSR
loci. The resulting F2:4 families were tested under two water
treatments. The authors identied 12 QTL, i.e. two for plant height,
ve for anthesis-silking interval, four for root mass, and one for grain
yield, however, most of them showed over-dominant gene action. In
the same crop, Guo et al. (2008) characterized QTL for some key traits
such as ower time, plant height, yield and yield components using
recombinant inbred lines developed from the cross 5003 p138 under
both water decit and well-watered regimes. They identied 51 QTL
for 10 traits on 10 different chromosomes. Under water decit
conditions, 22 QTL were found for 7 traits. Phenotypic variation linked
to each QTL ranged from 1.68 to 13.3%.
In wheat, the position of genes exhibiting a signicant effect on ABA
accumulation due to drought stress was identied using a series of single
chromosome substitution lines and populations obtained from a cross
between a high-ABA-producing cv. Ciano 67 and a low-ABA-producing
cv. Chinese Spring (Quarrie et al., 1994). They observed that chromosome 5A carries gene(s) for ABA accumulation. MAPMAKER-QTL
showed that the ABA quantitative trait locus is located between the
two loci Xpsr575 and Xpsr426, approximately 8 cM from Xpsr426.
In rice, for example, a number of drought-related QTL have been
identied for different growth and physiological traits involved in
drought tolerance (Latte et al., 2004). For example, Courtois et al.
(2003) found 28 QTL responsible for various root characteristics
172
Table 1
Drought tolerant cultivars/lines of different crops developed through conventional breeding at different centers/institutions.
Crop
Cultivar/line
How developed
Centers/institutions involved
Reference
Peanut (Arachis hypogaea

L. Fabaceae subsp.
hypogaea var. vulgaris )
ICGV 87354
SEA 5
Plant Materials Identication

Committee of the International
Crops Research Institute for the
Semi-Arid Tropics (ICRISAT), India
International Center for Tropical
Agriculture (CIAT), Cali, Colombia
Reddy et al. (2001)
Common bean
(Phaseolus vulgaris L.)
Derived from a cross between

Argentine and PI259747 and
developed through nine
generations of bulk selection
Developed from the interracial
double cross population BAT 477/San
Cristobal 83/Guanajuato 31/Rio Tibagi
Derived from double cross
population BAT 477San Cristobal
83/BAT 93/Jalo EEP 558
Developed from the single cross Red
Kloud ICA 10009
The complete pedigree of CO46348
was unknown, however it was derived
from a single cross with the
pinto cultivar Othello
Derived from a single plant selection
from F11 population
SEA 13
A 195
Line CO46348
Safower
(Carthamus tinctorius L.
Morlin
Chickpea
(Cicer arietinum L.)
Wheat
(Triticum aestivum L.)
FLIP 87-59C
Willow Creek
Ripper
NE01643
Prairie Red
Jinmai 50
Developed by crossing ILC3843 with

FLIP87
Through breeding in single replication
observation (SROB) nurseries
Developed by using a modied bulk
breeding procedure
A bulk breeding procedure was used
and approximately 50% of F3
population was visually selected on
the basis of agronomic appearance
Derived from the crosses and
backcrosses of CO850034/PI372129/5
TAM 107
Developed from the cross,
Pingyang181 Qingfeng1
Tall fescue
(Festuca arundinacea)
Nanryo
First breeder seed was produced from

a breeder's block composed of eight
clones
Soybean
[Glycine max (L.) Merr.]
R01-416F and
R01-581F
Wheatgrass
[Elymus trachycaulus
(Link) Gould ex Shinners]
FirstStrike
Barley
(Hordeum vulgare L.)
Lenetah
Both lines were originated after

selection from F9 population.
developed from a cross between
Jackson and KS4895
The source plants of FirstStrike
originated from seeds of three
germplasm collections i.e., [(5354
[N/S], 1516 [E/W]), (7172
[N/S], 1415 [E/W]) and 10 mi
Developed using a pedigree selection
procedure with all early generation
population and selected from the
cross, 94Ab12981 91Ab3148
Selected for drought resistance in
an F3 population received from
ICARDA, initially originating from a
single cross Baladi Bahteem/SD729Por 12762BC
The pedigree breeding method was
used for development and it was
originated from the cross between
the Egyptian local cultivar Giza 121
and the line 366/13/1 (Giza 117/
Bahteem 52//Giza 118/FAO 86)
Plant selection within superior F4
populations
Derived from an F3 population. The
pedigree method of breeding was
used and Giza 132 originated from
the cross Rihane-05//As46/Aths/3/Aths/
Lignee 686
Giza 126
Giza 2000
Giza 121, Line

366/13/1
Giza 132
Singh et al. (2001)
International Center for Tropical

Agriculture (CIAT), Cali, Colombia
Singh et al. (2001)
Centro Internacional de Agricultura

Tropical (CIAT), Palmira, Colombia
Colorado Agricultural Experiment
Station in cooperation with the
University of Idaho and USDA-ARS
Singh et al. (2007)
Eastern Agricultural Research

Center and Montana Agricultural
Experiment Station, Sidney
International Center for Agricultural
Research in the Dry Areas (ICARDA)
Montana Agricultural Experiment
Station, Sydney
Station, USA
Nebraska Agricultural Experiment
Station and the USDA-ARS
Brick et al. (2008)
Bergman et al. (2001)
Singh et al. (1996)

Cash et al. (2009)
Haley et al. (2007)
Baenziger et al. (2008)

Station, USA
Quick et al. (2001)
Wheat Breeding Innovation Group

(WBIG) in the Cotton Research
Institute of Shanxi Agri. Sci. Academy,
Yuncheng, China and
Testing and Appraising Committee
of Crop Cultivars of Shanxi
Province (TACCCSP)
Kyushu Okinawa National Agricultural
Research Station (KONARC),
Kumamoto, Japan and Japanese Grassland
Farming Forage Seed Association and
the USDA-ARS, Grazinglands
Research Laboratory, El Reno, OK USA.
Arkansas Agricultural Experiment
Station, USA
Xinglai et al. (2006)
USDA-ARS Forage and Range Research

Laboratory at Utah State University,
Logan, UT, in collaboration with
the U.S. Army Engineer Research and
Development Center, Hanover, NH
Agricultural Research Service,
Aberdeen, ID, in cooperation
with the Idaho Agricultural
Experimental Station
Research in the Dry Areas (ICARDA)
Jenson et al. (2007)
Kindiger et al. (2006)
Chen et al. (2007)
Obert et al. (2008)
Noaman et al. (1995)
Barley Research Department, Agricultural

Research Center at Giza, Egypt
Sakha Research Station Northern

Delta Region, Egypt
Barley Research Department, Agricultural
Research Center at Giza, Egypt, and
International Center for Agricultural Research
in the Dry Areas (ICARDA), Aleppo Syria

173
Table 1 (continued)
Crop
Cultivar/line
How developed
Centers/institutions involved
Reference
Lignee 686
Plant selections within superior rows

of F3 families were made and seeds
from these plants were grown as F4
plants after which only the best
appearing lines were combined
together and carried forward to the
next generation for yield tests
Developed from a single cross,
Baladi Bahteem SD729-Por 12762BC
West Asia and North Africa (WANA)

and Sakha Research Station in the
northern Delta Region of Egypt

Research in the
Dry Areas (ICARDA) Aleppo, Syria
Field Crops Development Centre (FCDC),
Lacombe, AB, Canada
Ohio State University (OSU), Ohio
Agricultural Research and development
Center, USA
Giza 126
Bentley
Maize (Zea mays L.)
Oh605
Sixteen tropically
adapted inbred
lines (TZEI 1 to
TZEI 16)
ND2005 and
ND2006
TZE-W Pop DT STR
C4 and TZE-Y Pop
DT STR C4
Derived from crossing I92125 with

TR229
22 selected full-sib progenies from
the AAE population
were intermated with 30 full-sib
progenies obtained
from OhS3267LAN plants
Developed from four diverse
germplasm sources with tolerance
to drought namely, TZE-W
Pop DT STR C0, TZE-Y Pop DT STR C0,
TZE Comp 5-Y C6, and TZE-W Pop
3 1368 STR C0
Developed through an integration
of recurrent selection
and pedigree breeding methods
Developed through backcrossing,
inbreeding, hybridization and
selection
involved in drought resistance. Similarly, 14 QTL related to osmotic

adjustment have been identied in an independent study on rice (Robin
et al., 2003). In another study pertaining to identication of QTL related
to root traits and osmotic adjustment in rice, 36 QTL related to some key
root traits and 5 related to osmotic adjustment were identied (Zhang et
al., 2001). While assessing the role and genetic mechanism of leaf water
potential (LWP) in japonica rice (Oryza sativa L. subsp. japonica) under
various water limited regimes of upland and lowland environments,
Yan-Ying et al. (2008) detected 6 QTL for LWP. Of the 6 QTL, the two for
LWP at predawn in upland (wpiu1 and wpiu4) and one for LWP at
midday in upland (wpu6) explained 5.4%, 7.9%, and 10.0% of the
phenotypic variation, respectively.
Despite the identication of QTL for growth or physiological
phenomena related to drought tolerance, QTL for yield and yield
components in rice have also been identied (Bernier et al., 2008). For
example, while carrying out the genetic analysis in a rice population
(CT9993IR62266) by molecular markers, Babu et al. (2003) identied 5
QTL related to drought tolerance in two eld experiments in southern
India explaining a maximum of 28% of the total genetic variation for rice
grain yield under water limited conditions. Using the same rice mapping
population, Lanceras et al. (2004) performed a trial in lowland conditions
in Thailand to identify drought-related QTL. They identied 4 QTL related
to grain yield under different water limited regimes, but none of these QTL
was similar to those earlier reported by Babu et al. (2003) in the same
population. Bernier et al. (2008) ascribed this difference in expression of
the QTL to the substantial QTLenvironment interaction, because a large
QTLenvironment interaction can minimize the repeatability of QTL.
Recently, Kamoshita et al. (2008) using the same rice mapping population
as previously used by Babu et al. (2003) and Lanceras et al. (2004)
identied a number of QTL for several drought-related traits including
deep rooting. They identied four vital genomic regions on chromosomes
1, 4, 8, and 9 wherein co-placed a number of QTL for different traits
involved in regulating grain yield under water limited conditions. They
anticipated the use of these genomic regions in marker-assisted selection
aimed for the development of drought tolerant rice cultivars/lines. Kumar
et al. (2007) identied a QTL on chromosome 1 near sd1 which accounted
for 32% of the total genetic variation for grain yield under drought
Juskiw et al. (2009)

Pratt and Casey (2006)
International Institute of Tropical

Agriculture (IITA), Ibadan, Nigeria
Badu-Apraku et al. (2006)
North Dakota Agricultural Experiment

Station, USA
Carena and Wanner (2009)
International Institute of Tropical

Agriculture (IITA), Ibaban, Nigeria
Badu-Apraku and Yallou (2009)
conditions, but only 4% under stress free conditions. This indicates that this
QTL accounts for variation in grain yield under stressful conditions, but not
for that under non-stress conditions. Its effect was also found to be consistent over years.
In view of a number of previous reports, it is evident that molecular
mapping and a number of QTL associated with drought tolerance
identied in different crops can be effectively used in appropriate
breeding programs meant for improving crop drought tolerance. In a
routine QTL mapping, complex phenotypes are reproduced based on the
available molecular information fundamental to the understanding of
genetic basis of stress tolerance. QTL mapping is thus an effective way of
detecting specic components that permit direct appraisal of stress
tolerance. However, due to the intricacy of genetic system and association
among genetic factors or even between genetic factors and the prevalent
environment, in most cases a limited number of stable QTL than the
expected have been detected across genetic pools and environments.
3.2. Manipulation of QTL for developing drought tolerant crop cultivars/lines
Recent molecular biology tools have undoubtedly led to the
development of DNA markers that have been effectively used to identify
QTL a number of traits in different crops. Furthermore, with the
development of detailed molecular linkage maps, a signicant progress
has been made during the last few years in marker-assisted selection
procedures that have allowed pyramiding of desirable traits to attain
substantial improvements in crop drought tolerance. Of different potential
crops, rice has been the most focused crop for improving its drought
tolerance through MAS. For example, Steele et al. (2006) conducted a
marker-assisted back-crossing (MABC) breeding program to improve
some key root morphological characteristics, involved in the drought
tolerance of an Indian upland elite rice variety, Kalinga III. Cultivar
Azucena, an upland japonica variety originally from the Philippines was
the donor parent. The authors targeted ve fragments on different
chromosomes for introgression, i.e., four contained QTL for improved root
morphological characteristics such as root length and thickness, and the
fth one had a recessive QTL for aroma. The selection carried out in three
backcross (BC) generations and two further crosses between BC3 lines
174
resulted in pyramiding all ve segments. It is imperative to note that

pyramiding of four root QTL was achieved after eight generations, using
3000 marker assays in 323 lines. The authors evaluated 22 near-isogenic
lines (NILs) for root traits in ve different eld trials in Bangalore, India. Of
the ve segments, the target segment on chromosome 9 (RM242
RM201) from cv. Azucena markedly improved root length under both
well-irrigated and water limited conditions. In a later study, Steele et al.
(2007) conducted a eld trial to test some key agronomic traits in nearisogenic lines (NILs) derived from the previous study (Steele et al., 2006).
Four NILs were evaluated in eld trials conducted in eastern and western
India for three years. All four NILs excelled Kalinga III in terms of grain and
straw yield. All these efforts using the marker-assisted breeding have
resulted in the release of a rst ever highly drought tolerant rice variety,
Birsa Vikas Dhan 111 (PY 84), in the Indian state of Jharkhand (Steele,
2009). Early maturity, high drought tolerance and high grain yield with
good grain quality are the prominent characteristics of this novel variety.
At IRRI, efforts have also been made to improve drought tolerance of rice
using the marker- assisted breeding approach. For example, Bernier et al.
(2007), while screening a population of 436 F3 lines derived from a cross
between two upland rice cultivars, Vandana and Way Rarem, selected
some lines. The selected lines were evaluated under water stress and nonstress conditions in some eld trials conducted for two years to identify
QTL involved in drought resistance. A QTL (qtl12.1) with a marked effect
on grain yield under drought stress was identied on chromosome 12 in
both years, which was derived from the susceptible parent, Way Rarem.
Under stress conditions, the QTL (qtl12.1) also improved biomass
production, harvest index, and plant height, while it showed reduced
number of days to owering. However, in contrast, under well-watered
treatment, the QTL did not affect any of the earlier mentioned characters.
According to Bernier et al. (2007), this is the rst QTL reported in rice
possessing a substantial and repeatable effect on grain yield production
under harsh eld drought conditions. To further conrm the effectiveness
of this QTL in improving drought tolerance in rice, Bernier et al. (2009)
have recently conducted vast eld trials at different locations, i.e., 10 at
IRRI and 11 in eastern India (Table 2). It is important to note that the
relative effect of the QTL on grain yield was augmented with the severity
of drought stress, and had no effect under well-irrigated regimes. This
conrms that the QTL, qtl12.1 has a pronounced and steady effect on grain
yield under upland water stress conditions, in variable environments.
Although pearl millet [Pennisetum glaucum (L.) R. Br.] is known for its
high drought tolerance, drought is a major constraint for its optimum
production in many areas of the world. Thus, breeding for water stress
tolerance in pearl millet is a major challenge for many research institutes.
The breeding research at ICRISAT, India, has resulted in mapping several
QTL for stover and grain yield under terminal water decit conditions
(Serraj et al., 2004). However, a preliminary evaluation of a putative
drought resistance QTL on linkage group 2 (LG 2) of pearl millet was
carried out by evaluating hybrids generated through topcross pollinators
bred from progenies derived from the original mapping population
comprising the tolerant allele at the target QTL. Thirty six topcross hybrids
were appraised in 21 different eld environments, wherein they were
subjected to control and drought-stressed regimes during the reproductive stages including owering and grain lling stages (Table 2). The
hybrids containing the QTL out-yielded under water stress regimes, but at
the cost of reduced yield under well-watered conditions. Subsequent
evaluations as reported by Serraj et al. (2005) were based on testcross
hybrids of drought tolerance QTL introgression lines in the gene pool of
the drought-sensitive parent of the mapping population, H 77/833-2.
These introgression lines were, in fact, developed by marker-assisted
backcrossing of a putative vital drought tolerance QTL into H 77/833-2
from the mapping population's drought tolerant parent. While evaluating
all QTL introgression lines under different moisture regimes, it was found
that many of the lines excelled the test cross hybrids in terms of grain yield
under terminal drought stress.
In another study with pearl millet, line 863B has been found to have a
superior general combining ability for the grain lling trait under
terminal drought conditions (Yadav et al., 2004). A mapping population

developed from a cross 863B ICMB 841 was evaluated under earlyand late-drought stress conditions, which led to the identication of two
genomic regions in line 863B associated with improved panicle harvest
index as well as with high drought tolerance (Yadav et al., 2004). The
genomic regions, Qgydt.icp-2.1 on LG 2 and Qgydt.icp-6.1 on LG 6
represent 23.6% and 14.4% of the total variation for panicle harvest
index, respectively. Subsequent appraisal of the mapping population
test crosses in different environments exhibited Qgydt.icp-2.1 as the
major QTL for marker-assisted selection program for drought tolerance
(Bidinger et al., 2007).
In cotton, QTL for yield and different drought-related secondary traits
such as carbon isotope ratio (13C), turgid solute potential, and leaf
chlorophyll content were exchanged between the potential cultivars of
the two cotton species G. barbadense (GB) cv. F-177 and G. hirsutum (GH)
cv. Siv'on through marker-assisted selection (Levi et al., 2009a,b). Several
of the resulting NILs out-performed in terms of the physiological traits for
which they were introgressed. In a subsequent study (Levi et al., 2009a,b),
photosynthetic efciency of two selected NILs and their recipient parents
were evaluated under water limited and well-watered eld conditions.
The GB NIL showed a stable rate of net CO2 assimilation rate under varying
leaf water potentials with a signicant superiority over its recipient
parent, F-177. The high net photosynthetic rate in this NIL was found to be
associated with lower stomatal limitation, higher activity of Rubisco and
higher rate of electron transport. In contrast, the other NIL (GH NIL)
exhibited higher mesophyll conductance under water limited conditions
than its recipient parent, Siv'on, but these genotypes had almost similar
values of net photosynthetic rate. However, both types of NILs did not
perform well in terms of yield relative to the recipient parents under
drought stress conditions.
A marker-assisted backcross (MABC) selection program meant for
improving grain yield under water limited conditions in tropical
maize was conducted at CIMMYT, Mexico (Ribaut and Ragot, 2006),
which involved the crossing of drought resistant line Ac7643 with a
drought susceptible line CML247. Marker-based selection was carried
out stepwise on all four generations (from BC1F1 to BC2F3). After the
four consecutive MABC cycles, the 70 BC2F3 individuals exhibiting the
closest allelic composition at target and non-target loci were bred
with two CIMMYT testers (CML254 and CML274). Thirty genotypes
were selected on the basis of their performance in terms of grain yield
and some key agronomic traits. However, the best ve MABC-derived
hybrids produced yield about 50% more than that of control hybrids,
but in contrast, under mild water stress, there was no difference
between MABC-derived hybrids and the control plants. This conrms
that the expression of genetic variation for drought tolerance mainly
depends on the severity of drought stress.
In barley, an attempt has been made to improve yield under
dryland conditions using wild barley (Hordeum spontaneum) as a
potential source of alleles for drought tolerance (Baum et al., 2003). A
population developed by backcrossing cultivated barley (Hordeum
vulgare) with H. spontaneum was evaluated in three Mediterranean
countries under rain-fed conditions to detect wild barley alleles
involved in producing high yield under drought conditions. Six QTL
from the wild barley were found to be responsible for enhanced yield
under water limited conditions. These results exhibit that identication of new alleles from wild relatives is a useful means of improving
drought tolerance in different potential crops.
Marker-assisted selection was also employed to improve the staygreen trait involved in the drought tolerance of sorghum (Harris et al.,
2007). Four major QTL (Stg1 to Stg4) contributing to the stay-green trait
were mapped using a population derived from BT642 and RT7000.
The genotype BT 642 is a potential source of stay-green trait.
Physiological evaluation of four RT7000 NILs comprising Stg1, Stg2,
Stg3, or Stg4 showed that BT642 alleles in each of these loci could
substantially contribute to the stay-green trait. However, RT7000 NILs
having BT 642 DNA relating to Stg2 showed higher stay-green
Table 2
Enhancing drought tolerance in different crop lines/varieties using marker-assisted selection.
QTL used
QTL donor
line/cultivar
Recipient line/
cultivar
Line/cultivar developed
Trait improved
Rice (Oryza sativa L.)
QTL9 (on chromosome 9)
Azucena
Kalinga III
QTL9 (on chromosome 9)
Azucena
Kalinga III
Near-isogenic lines
(NILs)
Near-isogenic lines
(NILs)
QTL2 (on chromosome 2) QTL9

(on chromosome 9) and QTL11
(on chromosome 11)
qtl12.1 (on chromosome 12)
Azucena
Kalinga III
Birsa Vikas Dhan 111

(PY 84)
Improved root length and thickness under both irrigated and

Steele et al. (2006)
drought-stressed eld conditions
Under eld conditions, NILs out-performed Kalinga III for grain
Steele et al. (2007)
and straw yield. However, the lines had higher straw yield than
grain yield as introgressed genes involved in partitioning of biomass
to the roots and stems, rather than to the grain
Early maturing and high yielding; good grain quality; drought tolerant Steele (2009)
Way Rarem
Vandana
F3-derived population
qtl12.1
LGA02, Chromosomes 06, 02,
and LGD05
Vandana
Gossypium
barbadense
(GB) cv. F-177
GH cv. Siv'on
F3-derived populations
BC3F3 NILs
Chromosomes 25, 22 and LGA06
Way Rarem
Gossypium
hirsutum (GH)
cv. Siv'on
GB cv. F-177
LGA02
GH cv. Siv'on
GB cv. F-177
NIL 14
Chromosome 25
GB cv. F-177
GH cv. Siv'on
NIL 3-2
PRLT 2/89-33
H-77/833-2
BC4F3-derived NILs
PRLT 2/89-33
PRLT 2/89-33
H77/833-2
H 77/833-2
863B
ICMB 841
F2 population
Topcross pollinators
(TCP) hybrids F4
population
F4 population
Cotton (Gossypium spp.)
Pearl millet
QTL on linkage group 2 (LG 2)
[Pennisetum glaucum (L.) R. Br.]
QTL on LG2
QTL on LG2
Qgydt.icp-2.1 on LG 2
Maize (Zea mays L.)
Five QTLs located on chromosomes Ac7643

1, 2, 3, 8, and 10
CML 247
Barley (Hordeum vulgare L.)
81 QTLs were used, out of

which 6 (1H-3, 2H-1, 3H-2, 4H-3,
1H-5, 3H-1 and 3H-4)
were for grain yield
Stg1, Stg2, Stg3, or Stg4
Hordeum
spontaneum
Hordeum
vulgare
BT 642
RT 7000
Sorghum
[Sorghum bicolor (L.) Moench]
BC3F3 NILs
Marker-Assisted Back
Cross (MABC)-derived
BC2F3 hybrids
Back Cross population
Near-Isogenic RT 7000
Reference
Improved biomass production; earlier owering; higher panicle

Bernier et al. (2009)
numbers and grain yield
Improved biomass production; panicle number; and 47% average yield Bernier et al. (2007)
High seed-cotton yield was negatively correlated with 13C, specic
Levi et al. (2009a)
leaf weight and chlorophyll
High yield was negatively correlated with 13C and osmotic potential
and positively with stomatal conductance, specic leaf weight and
chlorophyll
The GB NILs exhibited high seed-cotton yield, low osmotic potential,
high chlorophyll and lower leaf size
High seed-cotton yield and low osmotic potential, lower specic
leaf weight, higher stomatal density and smaller epidermal cells
Introgressed lines showed signicant positive general combining
ability for grain yield and out-yielded test cross hybrids
Improved grain yield and drought tolerance
Higher in yield in a series of both absolute and partial
terminal drought stress
Improved panicle harvest index (PNHI), a ratio of grain dry
mass/panicle dry mass and terminal drought tolerance
Under severe drought stress conditions MAS derived varieties
were about 50% better in yield and showed reduced asynchrony
between male and female owering
Improved grain yield, and reduced negative impact of
drought on grain lling
During eld trials they retained more green leaf area, delayed
leaf senescence and better grain yield at maturity under
drought stress conditions
Levi et al. (2009b)
Serraj et al. (2005)

Howarth and Yadav (2002)
Bidinger et al. (2005)
Crop
Yadav et al. (2004)

Ribaut and Ragot (2006)
Baum et al. (2003), Talame

et al. (2004), Tuberosa and
Salvi (2006)
Harris et al. (2007)
175
176
characteristic at maturity than that in RT7000 or the other RT7000

NILs under terminal drought stress. These NILs also had markedly lower
rates of leaf senescence with respect to that in RT7000. These results
clearly show that improvement in drought tolerance of sorghum is
possible through map-based cloning of the genes responsible for the vital
secondary traits such as stay-green or delayed leaf senescence.
All the above-mentioned reports clearly show that considerable
improvement in plant drought tolerance is possible through markerassisted selection. Marker-assisted selection, undoubtedly, allows to
pyramid genes at two or more loci to improve drought tolerance. The
identication of QTL for yield or secondary traits plays a key role in
improving drought tolerance in different crops through MAS. In fact,
when a marker-trait association has been found clearly, MAS can
minimize to a great extent the dependence on particular environmental
conditions during the selection procedure, one of the main barriers
encountered in the traditional breeding of traits affected by drought
stress (Tuberosa and Salvi, 2006). Although the achievements made so
far through MAS in improving drought tolerance seem to be simple and
straightforward, the main problem being faced by the breeders employing MAS is the challenge of accuracy and preciseness in QTL identication as well as the application of the knowledge resulting to a
successful MAS program. Considerable genetic environment interaction, the large number of genes controlling yield, and erroneous use of
mapping populations resulting from parents that have narrow difference
in drought tolerance, has adversely affected the programs entailing
mapping of QTL for high yield under drought stress. Restrictions of
molecular markers encountered previously have been exceeded with
the advent of gene-based numerous SNP markers. Using SNP and other
markers, high density genetic maps can be constructed for the detection
and characterization of QTL/genes responsible for drought tolerance. In
fact, with the accessibility of genome sequence information of each crop,
integrated genetic and physical maps and SNP markers for specic traits
will lead to a substantial role in molecular breeding for abiotic stress
tolerance including drought tolerance. In 2005, a rst attempt was made
to clone QTL (Salvi and Tuberosa, 2005), which is indeed an important
milestone in molecular breeding, having a substantial role to understand
and manipulate the traits responsible for drought tolerance (Tuberosa
and Salvi, 2006; Tondelli et al., 2006; Cattivelli et al., 2008).
4. Engineering crops for enhanced drought toleranceTransgenic
approach
The great challenge of food security being faced these days the worldover has directed plant scientists towards gene revolution after green
revolution due to advances in biotechnology. The gene revolution, in fact,
involves modication of qualitative and quantitative traits in an
organism by transferring desired genes from one species to another.
This strategy is referred to as the transgenic approach. In contrast to
classical breeding, the transgenic approach allows the incorporation of
only the specic cloned genes into an organism and restricts the transfer
of undesirable genes from donor organism. Through this approach,
pyramiding of genes with similar effects can also be achieved. Rapid
advance in recombinant-DNA technology and development of precise
and efcient gene-transfer protocols have resulted in efcient transformation and generation of transgenic lines in a number of crop species
(Gosal et al., 2009).
Transgenic approach is being pursued actively throughout the world
to improve traits including tolerance to biotic and abiotic stresses in a
number of crops (Ashraf et al., 2008). As with salt stress, plant responses to
drought stress are complex, because it involves many genes with additive
effects, so the prospects of improving drought tolerance in crops seem not
to be very bright. Despite this, efforts have been made during the last few
decades to generate transgenic lines of different crops, which have shown
improved tolerance to drought stress. Some of the transgenic lines so
produced for the over-expression of specic traits are listed in Table 3. The
major emphasis of bioengineers has been on engineering genes that
encode compatible organic osmolytes, plant growth regulators, antioxidants, heat-shock and late embryogenesis abundant proteins, and
transcription factors involved in gene expression.
It is now well established that compatible organic solutes play a
central role in plant drought tolerance (Ashraf and Foolad, 2007).
However, overproduction of compatible organic osmotica is one of the
prominent responses of plants exposed to osmotic stress (Serraj and
Sinclair, 2002; Ashraf et al., 2008) and the genes encoding the synthesis
of such organic solutes can be engineered to overproduce these solutes
in transgenic plants. For example, among the many organic osmolytes
known to play a substantial role in stress tolerance, glycine betaine (GB),
a quaternary ammonium compound, occurs richly in response to
dehydration stress (Mansour, 2000; Mohanty et al., 2002; Yang et al.,
2003; Ashraf and Foolad, 2007). However, for the biosynthesis of GB in
higher plants, choline monooxygenase (CMO) and betaine aldehyde
dehydrogenase (BADH) are two key enzymes. In some independent
studies with different crops, genes encoding these two enzymes have
been engineered (Table 3). For example, transgenic tobacco lines overexpressing CMO have been produced (Shen et al., 2002; Zhang et al.,
2008). These transgenic lines showed higher accumulation of glycine
betaine under water limited conditions and hence enhanced drought
tolerance. Similarly, a potential maize inbred line DH4866 was
transformed with the E. coli betA gene encoding choline dehydrogenase
(Quan et al., 2004). The transformed maize plants contained higher
levels of glycine betaine and showed higher tolerance to drought as
compared to wild-type plants when tested at the initial growth stages.
Like GB, proline is also an important compatible organic osmolyte
that plays a key role in stress tolerance. Pyrroline-5-carboxylate
synthetase (P5CR) is the key enzyme for proline biosynthesis. The
gene for this enzyme has been engineered in soybean (Ronde et al.,
2004), petunia (Yamada et al., 2005), and tobacco (Gubis et al., 2007). All
these transgenic lines showed enhanced accumulation of proline as well
as high drought tolerance (Table 3).
Trehalose, a nonreducing sugar, is also a potential organic osmoticum
which has a substantial role in the protection of plants against stresses.
However, transgenic lines of different crops have been generated using the
genes of some key enzymes involved in trehalose biosynthesis. For
example, enhanced drought tolerance has been achieved by transforming
the gene TPS1 for trehalose-6-phosphate synthase in tobacco (Romero
et al., 1997; Karim et al., 2007). Enhanced drought tolerance has also been
observed in transformed rice plants expressing chimeric gene Ubi1::TPSP
due to increased accumulation of trehalose (Jang et al., 2003). In these
studies and some other reported in the literature, engineering constitutive
over-expression of genes encoding TPS and/or TPP (trehalose-6-phosphate phosphatase) resulted in enhanced trehalose accumulation as well
as drought tolerance. However, the main problem with such transformation had been that it led to abnormal plant development under normal
growth conditions, because the gene transformed remained turned on all
the time. To resolve this problem, Wu and Garg (2003) alternatively
adopted another way to engineer enhanced trehalose accumulation in
such a manner that trehalose biosynthesis took place only when the plant
encountered abiotic stress. They employed a stress-inducible promoter for
the over-expression of E. coli trehalose biosynthesis genes (otsA and otsB)
as fusion gene (TPSP, trehalose-6-phosphate synthase phosphatase) for
developing abiotic stress tolerance in rice. It is pertinent to note here that
the TPSP fusion gene transformation resulted in normal growth under
non-stress conditions, but the expression of the fusion gene occurred only
under stress conditions. In another study, a TPS1TPS2 fusion gene
construct was incorporated into Arabidopsis thaliana through Agrobacterium using either the 35S or the stress regulated rd29A promoter (Miranda
et al., 2007). The lines over-expressing the TPS1TPS2 construct showed
normal growth as well as enhanced tolerance to multiple stresses such as
salinity, drought, freezing, and high temperature. However, in contrast,
the plants over-expressing TPS1 alone under the operation of 35S
promoter exhibited aberrant growth and form. From all these reports, it
is obvious that a substantial improvement in drought tolerance of plants
can be achieved through engineering the genes involved in trehalose

metabolism, without the occurrence of any cost in the form of abnormal
growth and development on growing the transgenic lines under normal
well-irrigated conditions.
Mannitol, a polyol, is one of the most important osmoprotectants that
play a vital role in plant stress tolerance. However, attempts have been
made to achieve improved drought tolerance by the over-expression of
mannitol in plants by engineering genes involved in the biosynthesis of
mannitol. For example, ecotypic expression of the mt1D gene (involved in
the biosynthesis of mannitol) in wheat plants increased tolerance to both
drought and salt stresses (Abebe et al., 2003). In contrast, tobacco plants
transformed with a mannitol-1-phosphate dehydrogenase gene resulted
in enhanced mannitol accumulation, but enhanced mannitol accumulation did not affect osmotic adjustment or drought tolerance in the
transformed plants as compared to those in the untransformed plants
(Karakas et al., 1997). In view of these contrasting reports, there is a need
to ascertain whether or not mannitol over-expression is related to drought
tolerance in different species. In case, there is a lack of relationship of
drought tolerance with mannitol accumulation, the efforts to engineer
crops for enhanced overproduction of mannitol and hence enhanced
drought tolerance would be futile.
Like other stresses, drought stress leads to increased accumulation of
reactive oxygen species (ROS) in plants thus causing an oxidative stress.
To counteract these ROS, plants can intrinsically develop different types of
antioxidants. Overproduction of antioxidants in response to droughtinduced oxidative stress has been found to be associated with the drought
stress tolerance of different plant species (Pastori and Foyer, 2002; Sunkar
et al., 2006). Furthermore, genes encoding different types of antioxidants
have been engineered in different plants for achieving enhanced drought
tolerance. For example, engineering of the gene SOD encoding superoxide
dismutase caused enhanced drought tolerance in alfalfa (McKersie et al.,
1996, 1997), potato (Perl et al., 1993), and rice (Wang et al., 2005).
Likewise, the gene for another potential antioxidant enzyme, ascorbate
peroxidase from Arabidopsis was over-expressed in tobacco chloroplasts
(Badawi et al., 2004). The transgenic lines so produced exhibited
enhanced tolerance to osmotic stress created by polyethylene glycol
(PEG). Transgenic lines of tobacco produced by over-expressing monodehydroascorbate reductase (MDAR) gene from Arabidopsis showed a
2.1-fold higher MDAR activity and 2.2-fold higher level of reduced
ascorbic acid than that in non-transformed plants (Eltayeb et al., 2007).
Transgenic plants showed enhanced resistance to ozone, salt, and PEG. Liu
et al. (2008) generated transgenic tobacco plants over-expressing VTE1
gene encoding tocopherol cyclase (VTE1), a key enzyme of tocopherol
biosynthesis. The transformed plants exhibited enhanced drought
tolerance which was associated with decreased electrolyte leakage, lipid
peroxidation and H2O2 content, but increased chlorophyll content
compared with the non-transformed plants.
Helicases, which catalyze the unwinding of DNA/RNA double helical
structures, are distributed in yeast, animals and plants (Luo et al., 2009).
These helicases comprising nine specic motifs are also referred to as
DEAD-box helicases depending on their highly conserved amino acid
sequence (Asp(D)Glu(E)Ala(A)Asp(D) in motif II (Gorbalenya and
Koonin, 1993; Tanner et al., 2003; Luo et al., 2009). Some earlier studies
have shown the putative role of helicases in plant abiotic stress tolerance
(Owttrim, 2006; Vashisht and Tuteja, 2006). Recently, Luo et al. (2009)
have isolated a DEAD-box-containing cDNA sequence from alfalfa
(Medicago sativa) and it was designated as M. sativa helicase 1 (MH1).
The ectopic expression of MH1 in Arabidopsis led to improved seed
germination and plant growth under drought, salinity, and oxidative
stress, which was found to be associated with enhanced capacity of
osmotic adjustment, activities ascorbate peroxidase and superoxide
dismutase, and proline content in the transgenic Arabidopsis plants. This
study clearly shows the signicant role of the over-expression of the
helicase, MH1, in drought tolerance by enhancing the capacity of plants to
counteract the reactive oxygen species (ROS) as well as adjust themselves
osmotically.
177
Late embryogenesis abundant (LEA) proteins may accumulate in

response to drought stress in plants and play a vital role in plant protection
against the adverse effects caused by drought stress (Hong et al., 2005;
Gosal et al., 2009). The putative role of LEA proteins in plant drought
tolerance has been suggested to be due to their involvement in the
maintenance of cell membrane structure and ion balance, binding of
water, and their action as molecular chaperones (Close, 1997; Browne
et al., 2002; Babu et al., 2004). However, efforts have been made during the
last two decades to engineer LEA genes for enhanced plant drought
tolerance. For example, engineering the LEA genes PMA1959 and PMA80
(encoding a group 1 LEA protein and a group 2 LEA protein, respectively;
both from wheat) in rice resulted in enhanced drought tolerance (Cheng
et al., 2002). Similarly, a LEA gene HVA1 (which encodes a group 3 LEA
protein) from barley was engineered in rice (Xu et al., 1996), and wheat
(Sivamani et al., 2000). Both rice and wheat transformed lines so produced
showed enhanced tolerance to drought stress. In two independent
studies, a LEA protein gene ME-lea N4 from Brassica napus was transferred
through Agrobacterium to lettuce (Lactuca sativa L.; Park et al., 2005a) and
Chinese cabbage (Brassica campestris. Pekinensis; Park et al., 2005b) using
the CaMV 35S promoter. The transgenic lines of both crops showed
enhanced tolerance to both salinity and drought. Recently, Dalal et al.
(2009) have assessed the role of a group 4 LEA protein, LEA4-1, from B.
napus in stress tolerance. Expression analysis showed that expression of
LEA4-1 gene in leaf tissues in Brassica species was induced by multiple
stresses including ABA, salinity, low temperature, and drought. However,
over-expression of BnLEA4-1 in Arabidopsis driven by the constitutive
CaMV 35S or stress-inducible RD29A promoter resulted in improved
tolerance of transgenic Arabidopsis to salinity and drought stresses.
Another LEA protein gene OsLEA 3-1 has been incorporated into rice via
Agrobacterium under the operation of different promoters (Xiao et al.,
2007). The rice transgenics developed particularly under the control of
constitutive CaMV 35S and stress-inducible HVA1 promoters showed
enhanced drought tolerance when tested under natural eld conditions
(Xiao et al., 2007). In view of all the above reports the prospective role of
LEA genes in protecting the plants from drought stress seems plausible.
Abscisic acid (ABA), a well known growth inhibitor, modulates a
number of key growth and physiological processes in plants, including
suppression in seed germination, maintenance of seed dormancy by
inhibiting cell growth, induction of stomatal closure thereby minimizing
transpiration to prevent water loss, and acceleration of abscission and
senescence (Finkelstein et al., 2002; Fujita et al., 2005). It is now evident
that ABA production is enhanced under water limited conditions and it
can effectively protect plants against drought stress (Shinozaki and
Yamaguchi-Shinozaki, 2000; Finkelstein et al., 2002; Xiong et al., 2002;
Fujita et al., 2005). In view of some earlier reports, it is apparent that many
of the drought stress-inducible genes detected in plants are activated by
ABA (Ingram and Bartels, 1996; Seki et al., 2002). A detailed examination
of the promoters of ABA-regulated genes has revealed a highly conserved
cis-acting ABA-responsive element (ABRE) (Giraudat et al., 1994; Busk
and Pages, 1998). Fujita et al. (2005) cloned three diverse cDNAs encoding
ABRE binding proteins (AREB1, AREB2, and AREB3) in Arabidopsis. They
also reported that the expression of AREB1 and AREB2 was up-regulated
by ABA, drought and salinity. They also showed that of the nine AREB
homologs reported in Arabidopsis, expression of the three members,
AREB1/ABF2, AREB2/ABF4, and ABF3/DPBF5, was stimulated by drought,
ABA, and high salinity in vegetative tissues. Furthermore, they showed
that the expression of only AREB1 gene was not sufcient to direct the
expression of downstream genes under non-stress conditions. However,
they overcame this problem by creating an activated form of AREB1
(AREB1DQT). It was interesting to note that AREB1DQT-over-expressing
Arabidopsis plants showed ABA hypersensitivity and improved drought
tolerance. However, AREB1 was found to be the most effective positive
regulator of ABA signaling in Arabidopsis under water decit conditions. In
other studies, over-expression of specic transcription factors such as the
dehydration-responsive element-binding protein 1A (DREB 1A), ABF3
and ABF4 caused enhanced drought tolerance genes in Arabidopsis plants,
178
Table 3
Improving plant drought tolerance through engineering genes for organic osmolytes, transcription factors, late embryogenesis proteins, and hormones.
Gene engineered
Transgenic host
Source organism
Trait improved
Reference
Beta vulgaris choline monooxygenase (BvCMO)
Tobacco (Nicotiana tabacum L.)
Beet (Beta vulgaris L.)
Zhang et al. (2008)
Choline dehydrogenase (betA)
Zea mays L. inbred line DH4866
Escherichia coli
Vigna aconitifolia
1-pyrroline-5-carboxylate reductase (P5CR)
Soybean (Glycine max cv. Ibis)
Arabidopsis thaliana L.
1-pyrroline-5-carboxylate synthetase
(P5CSF129A) and neomycine
phosphotransferase (nptII)
Triticum aestivum salt tolerance gene (TaSTRG)
Vigna aconitifolia
(Triticum aestivum L.)
Mannitol-1-phosphate dehydrogenase (mt1D)
Escherichia coli
Mannitol-1-phosphate dehydrogenase (mtlD)
Wheat (Triticum aestivum L.)
Escherichia coli
Pleurotus sajor-caju trehalose

phosphorylase (PsTP)
Pleurotus sajor-caju
Bifunctional trehalose-6-phosphate synthase or

phosphatase (TPSP) as a fusion gene of otsA
and otsB
Escherichia coli
Trehalose-6-phosphate synthase (TPS1)
Escherichia coli
Trehalose-6-phosphate synthase 1 and 2

(TPS1 and TPS2)
N-acetyl-L-glutamate synthase (SINAGS1)
Saccharomyces cerevisiae
Tomato (Lycopersicon esculentum L.)
Improved germination ability and higher accumulation of ornithine
Triticum aestivum ubiquitin 2 (Ta-Ub2)
Wheat (Triticum aestivum L.)
Faster germination; seedlings grew vigorously under water decit

conditions; improved CO2 assimilation rate of transgenic plants
Atriplex hortensis
Arabidopsis thaliana L. and
Oryza sativa L.)
Quan et al. (2004)
Shen et al. (2002)

Yamada et al. (2005)
Vendruscolo et al.
(2007)
Ronde et al. (2004)
Gubis et al. (2007)
Zhou et al. (2009)
Karakas et al. (1997)
Abebe et al. (2003)

Han et al. (2005)
Wu and Garg (2003)
Romero et al. (1997)
Karim et al. (2007)

Kalamaki et al.
(2009)
Guo et al. (2008)
Atriplex hortensis choline monoxygenase (AhCMO) Tobacco (Nicotiana tabacum L.)

Petunia (Petunia hybrida cv. Mitchell)
Arabidopsis thaliana and Oryza sativa
1-pyrroline-5-carboxylate synthetase
(AtP5CS or OsP5CS)
Triticum aestivum L. cv. CD200126
1-pyrroline-5-carboxylate synthetase (P5CS)
Higher accumulation of glycinebetaine in leaves, roots and seeds. Transgenic

line exhibited improved tolerance to toxic level of choline and drought stress
Higher accumulation of glycinebetaine; more tolerant to drought stress
at germination and
the young seedling stage. Most importantly, improved grain yield,
integrity of the cell
membrane and activities of enzymes under drought stress
Improved drought tolerance by accumulating high amount of glycinebetaine
Transgenic plants accumulated 1.52.6 times greater proline and
showed better growth
than wild-type plants under drought stress
Improved drought tolerance was mainly due to protection against
oxidative stress; proline
accumulation was high in transgenic plants
NADP+ levels decreased in wild type/antisense, while increased in
transgenic/sense plants. Sense plants accumulated highest
amount of praline
Transgenic plants accumulated high levels of proline and chlorophyll
content and were
better adapted to water stress
Improved plant survival rate, fresh weight, chlorophyll content, higher
proline, and soluble
sugar contents, and signicantly higher expression of putative proline
synthetase and
transporter genes than the non-transgenic plants
Increase non-structural carbohydrates, and mannitol accumulation but
did not affect osmotic
adjustment or drought tolerance
Improved fresh and dry weights, plant height, and ag leaf length in
transgenic plants
Transgenic plants showed normal growth, and better capacity to retain
water, while the wild
type and the only empty vector-transformed control plants withered severely
The genetically-engineered rice plants produced higher amounts
of trehalose, exhibited
sustained plant growth, less photo-oxidative damage, and more favorable
mineral balance under
drought stress conditions
Trehalose accumulating plants exhibited multiple phenotypic alterations
including stunted growth, lancet-shaped leaves, reduced sucrose content
and improved drought tolerance
Enhanced drought tolerance by water retention and root development
Table 3 (continued)
Gene engineered
Transgenic host
Source organism
R2R3 MYB transcription factor (MYB15)
Transcription factor (AP37) encoding

Oryza sativa cytochrome c gene (OsCc1)
Bacillus subtilis chimeric sacB
Ryegrass (Lolium multiorum Lam.)
Bacillus subtilis
Capsicum annuum xyloglucan

endotransglucosylase/hydrolase (CaXTH3)
Pepper (Capsicum. annuum L.

cv. Pukang)
Late embryogenesis abundant protein

HVA1 (HVA1)
Mulberry (Morus indica)
Late embryogenesis abundant (HVA1)
Abscisic acid-responsive element-binding

protein1 (AREB1)
Zea mays ABA/ water stress/ripening
induced protein (zm-Asr1)
Tocopherol cyclase VTE1 encoded by VTE1
Escherichia coli
Zea mays L.
Sorghum (Sorghum bicolor)
Medicago sativa helicase 1(MH1)
Alfalfa (Medicago sativa L.)
Receptor for activated C-kinase 1 (OsRACK1)
Rice (Oryza sativa L. subsp. japonica cv.

Nipponbare)
Ascorbate peroxidase (APX5)
Nicotiana tabacum L. cv. SR-1
Arabidopsis thaliana
Manganese superoxide dismutase (MnSOD)
Pea (Pisum sativum L.)
Boea hygrometrica late embryogenesis

abundant proteins (BhLEA1 and BhLEA2)
Boea hygrometrica
Reference
Zhao et al. (2007)
Bhatnagar-Mathur
et al. (2009)
Ding et al. (2009)
Oh et al. (2009)
Ye et al. (2001)
Cho and Hong

(2006)
Lal et al. (2008)
Babu et al. (2004)

Fujita et al. (2005)
Jeanneau et al.
(2002)
Liu et al. (2008)
Luo et al. (2009)
Dehydration-responsive element-binding protein Tall fescue (Festuca arundinacea Schreb.)

DREB1A/CBF3
Arabidopsis thaliana dehydration-responsive
Peanut (Arachis hypogaea L.)
element-binding protein (AtDREB1A)
Trait improved
both under drought and non-drought conditions
Transgenic plants showed increased resistance to drought and
had high proline accumulation
Enhanced activities of superoxide dismutase, ascorbate peroxidase,
and glutathione reductase
and enhanced proline level in the transgenic plants, while a dramatic
increase in the lipid
peroxidation was observed in the untransformed controls under water
limited conditions
Transgenic lines were hypersensitive to ABA in germination assays, more
susceptible to ABA-elicited inhibition of root elongation, and more sensitive to
ABA-induced stomatal
closure. The transcript levels of ABA biosynthesis, signaling and responsive
genes were generally higher in the seedlings of transgenic plants
than those in wild types
Improved grain lling rate and grain yield (1657%) and drought tolerance
of transgenic plants
Transgenic plants had a lower level of total fructose, unchanged
sucrose levels, and a
slight reduction in hexose levels. However, growth of the levan-accumulating
sacB-transgenic
plants was decreased with the onset of the reproductive phase
Transgenic plants exhibited abnormal leaf morphology; showed variable
twisting and bending along the edges, resulting in a severely wrinkled
leaf shape resulting into a
marked improvement in tolerance to severe water decit conditions
The transgenic plants showed better cellular membrane stability (CMS),
photosynthetic yield, less photo-oxidative damage and better water use
efciency under drought stress
Higher leaf relative water content, less reduction in plant growth,
better cell membrane protection and drought tolerance
AREB1 regulated novel ABRE-dependent
ABA signaling that enhanced drought tolerance in vegetative tissues
Transgenic plants showed increased intrinsic water use efciency accompanied
by a dry weight increase under drought conditions
Lower lipid peroxidation, electrolyte leakage and H2O2 content,
while higher chlorophyll and tocopherol contents in
transgenic plants as compared to wild type
Improved seed germination, plant growth, osmotic adjustment, and activities
of superoxide dismutase and ascorbate peroxidase
Reduced membrane peroxidation and production of malondialdehyde,
while enhanced activity of superoxide dismutase in transgenic rice plants.
RACK1 negatively regulated the redox system-related tolerance to drought stress
Enhanced net photosynthetic rate, 3.8-fold higher level of
APX activity, while reduced toxicity of H2O2 in transgenic plants
Reduced electrolyte leakage, injury, oxidative damage, while improved
photosynthetic rate,
SOD activity and drought tolerance
The relative water content of leaves and activities of photosystem II,
superoxide dismutase
and peroxidase increased, while membrane permeability decreased in
transgenic plants
Da-hong et al.
(2009)
Badawi et al. (2004)
Wang et al. (2005)
Liu et al. (2009)
179
180
which was found to be related to partial stomatal closure and decreased

transpiration (Kasuga et al., 1999; Joung-youn et al., 2002). Luchi et al.
(2001) have shown that the increase in one of the limiting reactions of
ABA biosynthesis with the precise gene paralog encoding At-NCED3, also
induced enhanced drought tolerance in Arabidopsis. In another study,
Jeanneau et al. (2002) examined the role of an ABA and drought regulated
maize gene ASR1 (Vienne et al., 1999) coding for an ABA stress ripening
protein, and the effect of photosynthesis regulation through the ectopic
expression of the S. bicolor C4-phosphoenolpyruvate carboxylase gene,
C4-PEPC, in transgenic maize. The transgenic maize lines so produced
showed enhanced photosynthetic capacity and water use efciency as
well as high biomass production under mild water decit conditions.
Recently, Ding et al. (2009) have shown that transgenic expression of
MYB15, encoding a transcription factor, R2R3 MYB, in Arabidopsis, showed
considerable sensitivity to exogenous ABA and enhanced tolerance to
both drought and salinity. The transgenic lines over-expressing MYB15
showed over-expression of genes involved in ABA biosynthesis (ABA1,
ABA2), signaling (ABI3), and responsive genes (AtADH1, RD22, RD29B,
AtEM6) after application of ABA, suggesting that improved stress tolerance
in the MYB15 transgenic lines is associated with enhanced expression of
the genes involved in ABA biosynthetic and signaling pathways as well as
those encoding the proteins involved in stress protection.
Transcription factors are specic types of proteins that bind DNA
and are involved in the regulation of gene transcription, hence gene
regulation. Since regulation of genes involved in stress tolerance is
important for improving this trait in plants, strenuous efforts are
being made these days to identify and characterize transcription
factors (regulatory proteins) involved in stress-specic gene regulation. However, several transcription factors have been identied,
which are involved in gene regulation in plants under water limited
conditions (Bartels and Sunkar, 2005; Vinocur and Altman, 2005). Of a
number of transcription factors listed elsewhere (Gosal et al., 2009),
dehydration-responsive element-binding factors (DREB) have
attracted the attention of many scientists since Jaglo-Ottosen et al.
(1998) and Liu et al. (1998) rst reported the up-regulation of many
genes in DREB1/CBF transgenic Arabidopsis involved in tolerance to a
variety of stresses including drought, salinity, freezing etc. Similarly,
transgenic Arabidopsis plants over-expressing DREB1/CBF3 operated
by the constitutive promoter CaMV 35S also exhibited improved
tolerance to salinity, drought and freezing (Kasuga et al., 1999).
Introduction of DREB1A into wheat driven by rd29A promoter resulted
in enhanced drought tolerance (Pellegrineschi et al., 2004). Ecotypic
expression of Arabidopsis DREB1A /(CBF3) into transformed rice plants
under the operation of constitutive promoter CaMV 35S, resulted in
improved tolerance to drought and salinity (Oh et al., 2005).
Dubouzet et al. (2003) isolated four rice CBF/DREB1A orthologs, OsDREB1A, OsDREB1B, OsDREB1C and OsDREB1D. However, the transgenic rice plants over-expressing OsDREB1 exhibited improved
tolerance to drought, salinity and freezing. In maize, over-expression
of ZmDREB2A under the control of constitutive or stress-inducible
promoter resulted in enhanced drought tolerance in plants (Qin et al.,
2007). Similarly, peanut plants transformed with rd29A:DREB1A had
higher transpiration efciency than the wild type under drought
stress (Bhatnagar-Mathur et al., 2007). In a recent study, BhatnagarMathur et al. (2009) have generated transgenic plants of peanut overexpressing Arabidopsis AtDREB1A, driven by a stress-inducible promoter, Atrd29A. Although the transgenic peanut plants accumulated considerably higher levels of some key antioxidant enzymes (superoxide
dismutase, ascorbate peroxidase, and glutathione reductase) and proline
content, and lower levels of lipid peroxidation as compared to those in the
wild-type plants under moisture stress conditions, all these antioxidant
and biochemical indicators had no signicant relationship with transpiration efciency of the transgenic plants over-expressing AtDREB1A.
Despite DREB, other transcription factors are also known to be
involved in plant stress responses. One such type is that with APETELA2
(AP2)-domain. Recently, Oh et al. (2009) have identied 42 AP2 genes in
rice, which are triggered by various stresses including salinity, drought,

freezing, and ABA. While carrying out the phylogenic analysis of these 42
stress-inducible AP2 genes, they have identied 6 subgroups (IVI) with
conspicuous signature motifs, and two genes, AP37 and AP59, belonging to
subgroups I and II, respectively, were triggered just after 2 h of exposure to
water decit and high saline conditions, but they differed in their
expression prole, particularly under ABA and low temperature. The
transformed rice plants over-expressing AP37 and AP59 under the
operation of the constitutive promoter OsCc1 showed enhanced resistance
to high drought and saline conditions at the vegetative growth stage.
However, as compared with OsCc1:AP59 plants, the OsCc1:AP37 plants
showed considerably higher tolerance to drought by producing 1657%
more grain yield over non-transgenic controls under severe drought
conditions of the eld. These ndings suggest the potential role of the
AP37 gene to improve drought tolerance in rice.
From the whole preceding discussion it is not hard to infer that
considerable progress can be made within the shortest possible time in
improving plant drought tolerance by engineering the genes involved in
the synthesis of organic osmolytes, plant growth regulators, antioxidants,
late embryogenesis abundant proteins, and transcription factors (regulatory proteins) involved in gene expression. However, most of the
transgenic lines of different crops were tested under controlled laboratory
or glasshouse conditions wherein they have shown a remarkable performance under simulated stress conditions. With the exception of only
two studies (rice transgenics, Xiao et al., 2007; Oh et al., 2009) none of the
transgenic lines produced through genetic engineering in different studies
has been tested in natural eld conditions. Thus, it is not known how they
would perform during eld testing, because a natural eld encounters a
myriad of environmental factors other than the drought stress.
The other key issue is that like salt tolerance, the degree of drought
tolerance varies with growth and development in most plant species (ElFar and Allan, 1995; Reddy et al., 2004; Rassaa et al., 2008). Thus, the
degree of drought tolerance observed in the transgenic lines at one
particular stage particularly at the initial growth stage, may not reect the
same when tested at other growth stages or as adult. So, there is a need to
ensure the overall drought tolerance of a plant species for farmers'
standpoint.
Like in the case of plant salt tolerance (Ashraf and Akram, 2009), most
of the drought tolerant transgenic lines of different crops developed are
based on only a single gene transformation, whereas the claims of the
scientists regarding the performance of the lines with respect to drought
tolerance seem to be overstated as earlier reported in the case of salt
tolerance (Flowers, 2004; Ashraf and Akram, 2009). Thus, manipulation of
a number of genes predominantly involved in stress tolerance to
transgenic plants seems to be a plausible approach. This will certainly
allow pyramiding of desirable traits to achieve considerable advance in
crop drought tolerance.
5. Conclusions and future challenges
The main focus of the present review has been on three prominent
plant breeding approaches for achieving enhanced crop drought tolerance,
i.e., conventional breeding, marker-assisted breeding, and genetic
engineering. Despite the fact that conventional breeding has many
limitations as listed elsewhere (Ashraf, 1994; Flowers, 2004; Ashraf and
Akram, 2009), a reasonable number of cultivars/lines tolerant to drought
stress have so far been developed, some of which have not only performed
well under controlled environmental conditions, but also under natural
drought-prone environments. Certainly, the traditional protocols employed in developing such drought tolerant cultivars/lines have not been
very cost-intensive, if we compare the cost being incurred these days on
modern plant breeding approaches including marker-assisted breeding
and genetic engineering. However, the main debacle with traditional
plant breeding approach is that it does not offer labor and time savings.
Marker-assisted breeding approach is a prospective alternative to
traditional breeding, because of being less time-consuming and labor-
and cost-effective. Molecular mapping and analysis of QTL have been

carried out for a number of qualitative and quantitative traits including
stress tolerance, which has undoubtedly resulted in a great magnitude
of knowledge and better understanding of the causal genetic phenomena that regulate these traits. However, limited success has resulted by
using this knowledge to manipulate genes in an effective way for the
improvement of a specic trait in crops, although some crop cultivars
developed through using these tools have performed extremely well
under eld stress conditions. However, there are a variety of reasons for
the limited success in terms of achieving enhanced drought tolerance
using the marker-assisted breeding approach. For example, a major
difculty confronting the scientists is the challenge of precise QTL
identication. In addition, a substantial genetic environment interaction, inconsistent repeatability, large number of genes regulating yield,
and invalid use of mapping populations have hampered the pursuits
involving mapping of QTL for enhanced drought stress tolerance.
Despite these, other factors also impede the application of QTL for
genetic improvement of a trait. For example, due to unfavorable
epistatic interaction, it is hard to transfer the effects of a desired allele to
an elite background material (Podlich et al., 2004; Collins et al., 2008).
Furthermore, in some cases, QTL from a specic background do not
show signicant effects or cease completely in different backgrounds,
even under analogous growth conditions (Cho and Hong, 2006; Collins
et al., 2008). This makes the utilization of QTL more intricate.
Transformation of the knowledge acquired from QTL-oriented molecular studies into a well-dened upshot for the stakeholders is one of the
key challenges confronting the breeders. Certainly, a multidisciplinary
approach including more specically the identication of QTL mediated
signal transduction in response to stresses needs to be adopted.
Furthermore, although QTL cloning procedures are unwieldy, they can
lead to a meaningful outcome because a cloned QTL can offer a reliable
marker for MAB as well as furnish an outline for the detection of superior
allelic variants in crop species (Till et al., 2007).
Genetic engineering (Transgenic approach) offers a promise whereby
one can expect a substantial improvement in a desired trait within the
shortest stretch of time. Engineering genes encoding organic osmolytes,
plant growth regulators, antioxidants, late embryogenesis abundant proteins, and transcription factors has resulted into transgenic lines which
have out-performed under controlled stress conditions. In most cases the
claims made by the researchers in terms of performance of transgenic
lines tested under controlled conditions, are considerably inated.
However, with the exception of a very few notable cases, most of the
transgenic lines of different crops have been rarely eld-tested. Thus, how
far these transgenic lines perform under eld stress conditions is not
known, because under natural eld conditions a line/cultivar has to face a
multitude of environmental factors other than the drought stress. Thus,
appraisal of performance of a transgenic line under natural eld conditions is mandatory for the stakeholders' perspective. It is also imperative
to note that most of the drought tolerant transgenic lines have been
developed using a single gene transformation, which may not be as
productive as if it had been developed using transformation of many
genes. Thus, transferring a number of prominent genes effectively involved in stress tolerance to transgenic plants seems to be a logical
approach. Although a large number of genes appear to be involved in
stress tolerance and most of them have been fully characterized, the
function of many of them in the mechanism of stress tolerance is yet to be
investigated.
References
Abebe T, Guenzi AC, Martin B, Cushman JC. Tolerance of mannitol-accumulating
transgenic wheat to water stress and salinity. Plant Physiol 2003;131:174855.
Ashraf M. Breeding for salinity tolerance in plants. Crit Rev Plant Sci 1994;13:1742.
Ashraf M, Akram NA. Improving salinity tolerance of plants through conventional
breeding and genetic engineering: an analytical comparison. Biotechnol Adv
2009;27:74452. doi:10.1016/j.biotechadv.2009.05.026.
Ashraf M, Foolad MR. Roles of glycinebetaine and proline in improving plant abiotic
stress resistance. Environ Exp Bot 2007;59:20616.
181
Ashraf M, Athar HR, Harris PJC, Kwon TR. Some prospective strategies for improving
crop salt tolerance. Adv Agron 2008;97:45-110.
Asins MJ. Present and future of quantitative trait locus analysis in plant breeding. Plant
Breed 2002;121:28191.
Babu RC, Nguyen BD, Chamarerk V, Shanmugasundaram P, Chezhian P, Jeyaprakash P, et al.
Genetic analysis of drought resistance in rice by molecular markers: association between
secondary traits and eld performance. Crop Sci 2003;43:145769.
Babu RC, Zhang J, Blum A, Ho THD, Wu R, Nguyen HT. HVA1, a LEA gene from barley
confers dehydration tolerance in transgenic rice (Oryza sativa L.) via cell membrane
protection. Plant Sci 2004;166:85562.
Badawi GH, Kawano N, Yamauchi Y. Over-expression of ascorbate peroxidase in
tobacco chloroplasts enhances the tolerance to salt stress and water decit. Physiol
Plant 2004;121:2318.
Badu-Apraku B, Yallou CG. Registration of striga-resistant and drought tolerant tropical
early maize populations TZE-W Pop DT STR C4 and TZE-Y Pop DT STR C4. J Plant
Registr 2009;3(1):8690.
Badu-Apraku B, Menkir A, Kling JG, Fakorede MAB. Registration of 16 striga resistant
early maturing tropical maize inbred lines. Crop Sci 2006;46:14101.
Baenziger PS, Beecher B, Graybosch RA, Ibrahim AMH, Baltensperger DD, Nelson LA, et al.
Registration of NEO1643 wheat. J Plant Registr 2008;2(1):3642.
Bnziger M, Setimela PS, Hodson D, Vivek B. Breeding for improved drought tolerance in
maize adapted to southern Africa. Proceedings of the 4th International Crop Science
Congress. Brisbane, Australia: Published on CDROM; 2004. 26 Sep 1 Oct.
Bartels D, Sunkar R. Drought and salt tolerance in plants. Crit Rev Plant Sci 2005;24:2358.
Baum M, Grandol S, Backes G, Jahoor A, Sabbagh A, Ceccarelli S. QTLs for agronomic
traits in the Mediterranean environment identied in recombinant inbred lines of
the cross Arta H. spontaneum 41-1. Theor Appl Genet 2003;107:121525.
Bergman JW, Riveland NR, Flynn CR, Carlson GR, Wichman DM. Registration of Morlin
safower. Crop Sci 2001;41:1640.
Bernier J, Kumar A, Venuprasad R, Spaner D, Atlin G. A large-effect QTL for
grain yield under reproductive-stage drought stress in upland rice. Crop Sci
2007;47:50718.
Bernier J, Kumar A, Serraj R, Spaner D, Atlin G. Review: breeding upland rice for drought
resistance. J Sci Food Agric 2008;88:92739.
Bernier J, Serraj R, Kumar A, Venuprasad R, Impa S, Gowdaa RPV, et al. The large-effect
drought-resistance QTL qtl12.1 increases water uptake in upland rice. Field Crops
Res 2009;110:3946.
Bhatnagar-Mathur P, Reddy DS, Lavanya M, Yamaguchi-Shinozaki K, Sharma KK. Stressinducible expression of Arabidopsis thaliana DREB1A in transgenic peanut (Arachis
hypogaea L.) increases transpiration efciency under water-limiting conditions.
Plant Cell Rep 2007;26:207182.
Bhatnagar-Mathur P, Devi MJ, Vadez V, Sharma KK. Differential antioxidative responses in
transgenic peanut bear no relationship to their superior transpiration efciency under
drought stress. J Plant Physiol 2009;166(11):120717. doi:10.1016/j.jplph.2009.01.001.
Bidinger FR, Serraj R, Rizvi SMH, Howarth C, Yadav RS, Hash CT. Field evaluation of drought
tolerance QTL effects on phenotype and adaptation in pearl millet [Pennisetum glaucum
(L.) R. Br.] topcross hybrids. Field Crops Res 2005;94(1):1432.
Bidinger FR, Nepolean T, Hash CT, Yadav RS, Howarth CJ. Identication of QTLs for grain
yield of pearl millet (Pennisetum glaucum (L.) R. Br.) in environments with variable
moisture during grain lling. Crop Sci 2007;47:96980.
Bowers JE, Abbey C, Anderson S, Chang C, Draye X. A high-density genetic
recombination map of sequence-tagged sites for Sorghum, as a framework for
comparative structural and evolutionary genomics of tropical grains and grasses.
Genetics 2003;165:36786.
Brick MA, Ogg JB, Singh SP, Schwartz HF, Johnson JJ, Pastor-Corrales MA. Registration of
drought-tolerant, rust-resistant, high-yielding pinto bean germplasm line
CO46348. J Plant Registr 2008;2(2):1204.
Browne J, Tunnacliffe A, Burnell A. Anhydrobiosis-plant desiccation gene found in a
nematode. Nature 2002;416:38.
Busk PK, Pages M. Regulation of abscisic acid induced transcription. Plant Mol Biol
1998;37:42535.
Carena MJ, Wanner DW. Development of genetically broad-based inbred lines of maize
for early-maturing (70-80RM) hybrids. J Plant Registr 2009;3:10711.
Cash SD, Bruckner PL, Wichman DM, Kephart KD, Berg JE, Boyner R, et al. Registration of
Willow Creek' forage wheat. J Plant Registr 2009;3(2):18590.
Cattivelli L, Rizza F, Badeck FW, Mazzucotelli E, Francia AMEM, Mare AT, et al. Drought
tolerance improvement in crop plants: an integrated view from breeding to genomics.
Field Crops Res 2008;105:1-14.
Chen M, Wang QY, Cheng XG, Xu ZS, Li LC, Ye XG, et al. GmDREB2, a soybean DREbinding transcription factor, conferred drought and high-salt tolerance in
transgenic plants. Biochem Biophys Res Commun 2007;353:299305.
Cheng Z, Targolli J, Huang X, Wu R. Wheat LEA genes, PMA80 and PMA1959, enhance
dehydration tolerance of transgenic rice (Oryza sativa L.). Mol Breed 2002;10:7182.
Cho EK, Hong ChB. Over-expression of tobacco NtHSP70-1 contributes to drought-stress
tolerance in plants. Plant Cell Reports 2006;25:34958.
Close TJ. Dehydrins: a commonality in the response of plants to dehydration and low
temperature. Physiol Plant 1997;100:2916.
Collins NC, Tardieu F, Tuberosa R. Quantitative trait loci and crop performance under
abiotic stress: where do we stand? Plant Physiol 2008;147:46986.
Concept Note, Combining breeding and biotechnology to develop water efcient maize
for Africa (WEMA). Afr Agric Technol Foundation.
Courtois B, Shen L, Petalcorin W, Carandang S, Mauleon R, Li Z. Locating QTLs controlling
constitutive root traits in the rice population IAC 165-Co39. Euphytica 2003;134:
33545.
Da-hong L, Hui L, Yang YL, Ping-ping Z, Jian-sheng L. Down-regulated expression of RACK1
gene by RNA interference enhances drought tolerance in rice. Rice Sci 2009;16(1):1420.
182
Dalal M, Tayal D, Chinnusamy V, Bansala KC. Abiotic stress and ABA-inducible group 4 LEA
from Brassica napus plays a key role in salt and drought tolerance. J Biotechnol 2009;139:
13745.
Ding Z, Li S, An X, Liu X, Qin H, Wang D. Transgenic expression of MYB15 confers enhanced
sensitivity to abscisic acid and improved drought tolerance in Arabidopsis thaliana.
J Genet Genomics 2009;36:1729.
Dubouzet JG, Sakuma Y, Ito Y, Kasuga M, Dubouzet EG, Miura S, et al. OsDREB genes in
rice, Oryza sativa L., encode transcription activators that function in drought, highsalt- and cold-responsive gene expression. Plant J 2003;33:75163.
El-Far IA, Allan AY. Responses of some wheat cultivars to sowing methods and drought
at different stages of growth. Assuit J Agric Sci 1995;26(1):26777.
Eltayeb AE, Kawano N, Badawi GH, Kaminaka H, Sanekata T, Shibahara T, et al.
Overexpression of monodehydroascorbate reductase in transgenic tobacco confers
enhanced tolerance to ozone, salt and polyethyleneglycol stresses. Planta 2007;225
(5):125564.
Falconer DS. Introduction to quantitative genetics. London New York: Longman; 1989.
Feng-ling FU, Zhi-Lei F, Shi-bing G, Shu-feng Z, Wan-chen L. Evaluation and quantitative
inheritance of several drought-relative traits in maize. Agric Sci China 2008;7(3):28090.
Finkelstein R, Gampala S, Rock C. Abscisic acid signaling in seeds and seedlings. Plant
Cell 2002;14:1545.
Flowers TJ. Improving crop salt tolerance. J Exp Bot 2004;55:30719.
Fujita Y, Fujita M, Satoh R, Maruyama K, Parvez MM, Seki M, et al. AREB1 is a transcription
activator of novel ABRE dependent ABA signaling that enhances drought stress tolerance
in Arabidopsis. Plant Cell 2005;17:347088.
Giraudat J, Parcy F, Bertauche N, Gosti F, Leung J, Morris PC, et al. Current advances in
abscisic acid action and signalling. Plant Mol Biol 1994;26:155777.
Gorbalenya AE, Koonin EV. Helicases: amino acid sequence comparisons and structure
function relationships. Curr Opin Struct Biol 1993;3:41929.
Gosal SS, Wani SH, Kang MS. Biotechnology and drought tolerance. J Crop Improvement
2009;23:1954.
Gubis J, Vakov R, erven V, Dragov M, Hudcovicov M, Lichtnerov H, et al.
Transformed tobacco plants with increased tolerance to drought. South Afr J Bot 2007;73:
50511.
Guo O, Zhang J, Gao Q, Xing S, Li F, Wang W. Drought tolerance through over-expression
of mono ubiquitin in transgenic tobacco. J Plant Physiol 2008;165:174555.
Haley SD, Johnson JJ, Peairs FB, Quick JS, Stromberger JA, Clayshulte SR, et al. Registration of
Ripper wheat. J Plant Registr 2007;1:16.
Han SE, Park SR, Kwon HB, Yi BY, Lee GB, Byun MO. Genetic engineering of droughtresistant tobacco plants by introducing the trehalose phosphorylase (TP) gene from
Pleurotus sajor-caju. Plant Cell Tissue Organ Cult 2005;82:1518.
Harris K, Klein R, Mullet J. Sorghum stay-green QTL individually reduces post-owering
drought-induced leaf senescence. J Exp Bot 2007;58:32738.
Hong BS, Zong-Suo L, Ming-An S. LEA proteins in higher plants: structure, function,
gene expression and regulation. Colloids Surf B Biointerf 2005;45:1315.
Howarth CJ, Yadav RS. Successful marker assisted selection for drought tolerance and
disease resistance in pearl milletIGER Innovations; 2002.
Humphreys MO, Humphreys MW. Breeding for stress resistance: general principles. In:
Ashraf M, Harris PJC, editors. Abiotic stresses: plant resistance through breeding
and molecular approaches; 2005. p. 1946.
Ingram J, Bartels D. The molecular basis of dehydration tolerance in plants. Ann Rev
Plant Physiol Plant Mol Biol 1996;47:377403.
Jaglo-Ottosen KR, Gilmour SJ, Zarka DG, Schabenberger O, Thomashow MF. Arabidopsis
CBF1 overexpression induces COR genes and enhances freezing tolerance. Science
1998;280:1046.
Jang IC, Oh SJ, Seo JS, Choi WB, Song SI, Kim CH, et al. Expression of a bifunctional fusion of
the Escherichia coli genes for trehalose-6-phosphate synthase and trehalose-6phosphate phosphatase in transgenic rice plants increases trehalose accumulation
and abiotic stress tolerance without stunting growth. Plant Physiol 2003;131: 51624.
Jeanneau M, Gerentes D, Foueillassar X, Zivy M, Vidal J, Toppan A, et al. Improvement of
drought tolerance in maize: towards the functional validation of the Zm-Asr1 gene and
increase of water use efciency by over-expressing C4-PEPC. Biochimie 2002;84:
112735.
Jenson KB, Palazzo AJ, Waldron BL, Bushman BS. Registration of FirstStrike Slender
wheatgrass. J Plant Registr 2007;1:245.
Joung-youn K, Hyung-in C, Min-young I, Soo-young K. Arabidopsis basic leucine zipper
proteins that mediate stress-responsive abscisic acid signaling. Plant Cell 2002;4:
34357.
Juskiw PE, Helm JH, Oro M, Nyachiro JM, Salmon DF. Registration of Bentley barley.
J Plant Registr 2009;3(2):11923.
Kalamaki MS, Alexandrou D, Lazari D, Merkouropoulos G, Fotopoulos V, Pateraki I, et al.
Over-expression of a tomato N-acetyl-L-glutamate synthase gene (SlNAGS1) in
Arabidopsis thaliana results in high ornithine levels and increased tolerance in salt
and drought stresses. J Exp Bot 2009;60(6):185971.
Kamoshita A, Babu CR, Boopathi NM, Fukai S. Phenotypic and genotypic analysis of
drought-resistance traits for development of rice cultivars adapted to rainfed
environments. Field Crops Res 2008;109(103):1-23. doi:10.1016/j.fcr.2008.06.010.
Karakas B, Ozias-Akins P, Stushnoff C, Suefferheld M, Rieger M. Salinity and drought tolerance
of mannitol-accumulating transgenic tobacco. Plant Cell Environ 1997;20:60916.
Karim S, Aronsson H, Ericson H, Pirhonen M, Leyman B, Welin B, et al. Improved drought
tolerance without undesired side effects in transgenic plants producing trehalose. Plant
Mol Biol 2007;64:37186.
Kasuga M, Liu Q, Miura S, Yamaguchi-Shinozaki K, Shinozaki K. Improving plant drought, salt
and freezing tolerance by gene transfer of a single stress-inducible transcription factor.
Nat Biotechnol 1999;17:28791.
Kindiger M, Gaub H, Hasegawac M, Katsurab Y, Ueyamad K, Gotob S, et al. Registration
of Nanryo tall fescue. Crop Sci 2006;46:18156.
Kong I, Dong J, Hart GE. Characteristics, linkage map positions, and allelic differentiation
of Sorghum bicolour (L.) Moench by DNA simple-sequence repeats (SSRs). Theor
Appl Genet 2000;101:43848.
Kumar R, Venuprasad R, Atlin GN. Genetic analysis of rainfed lowland rice drought
tolerance under naturally-occurring stress in eastern India: heritability and QTL
effects. Field Crops Res 2007;103:4252.
Latte HR, Price AH, Courtois B. Yield response to water decit in an upland rice
mapping population: associations among traits and genetic markers. Theor Appl
Genet 2004;109:123746.
Lal S, Gulyani V, Khurana P. Overexpression of HVA1 gene from barley generates tolerance to
salinity and water stress in transgenic mulberry (Morus indica). Transgenic Res 2008;17
(4):65163.
Lanceras J, Pantuwan G, Jongdee B, Toojinda T. Quantitative trait loci associated with
drought tolerance at reproductive stage in rice. Plant Physiol 2004;135:38499.
Levi A, Ovnat L, Paterson AH, Saranga Y. Photosynthesis of cotton near-isogenic lines
introgressed with QTLs for productivity and drought related traits. Plant Sci 2009a;177:
8896.
Levi A, Paterson AH, Barak V, Yakir D, Wang B, Chee PW, et al. Field evaluation of cotton
near-isogenic lines introgressed with QTLs for productivity and drought related
traits. Mol Breed 2009b;23:17995.
Liu Q, Kasuga M, Sakuma Y, Abe H, Miura S, Yamaguchi- Shinozaki K, et al. Two
transcription factors, DREB1 and DREB2, with an EREBP/AP2 DNA binding domain
separate two cellular signal transduction pathways in drought-and low-temperature-responsive gene expression, respectively in Arabidopsis. Plant Cell 1998;10:
1391406.
Liu X, Hua X, Guo J, Qi D, Wang L, Liu Z, et al. Enhanced tolerance to drought stress in
transgenic tobacco plants overexpressing. Biotechnol Lett 2008;30:127580.
Liu X, Wanga Z, Wanga L, Wua R, Phillips J, Deng X. LEA 4 group genes from the
resurrection plant Boea hygrometrica confer dehydration tolerance in transgenic
tobacco. Plant Sci 2009;176:908.
Luchi S, Kobayashi M, Taji T, Naramoto M, Seki M, Kato T, et al. Regulation of drought
tolerance by gene manipulation of 9-cis-epoxycarotenoid dioxygenase, a key
enzyme in abscisic acid biosynthesis in Arabidopsis. Plant J 2001;27:32533.
Luo Y, Liu YB, Dong YX, Gao XQ, Zhang YS. Expression of a putative alfalfa helicase increases
tolerance to abiotic stress in Arabidopsis by enhancing the capacities for ROS scavenging
and osmotic adjustment. J Plant Physiol 2009;166:38594.
MacLean JL, Dawe DC, Hardy B, Hettel GP. Rice Almanac: sourcebook for the most important
economic activity on Earth3rd ed. Wallingford, England: CABI Publishing; 2002.
Mansour MMF. Nitrogen containing compounds and adaptation of plants to salinity
stress. Biol Plant 2000;43:491500.
McKersie BD, Bowley SR, Harjanto E, Leprince O. Water-decit tolerance and eld
performance of transgenic alfalfa overexpressing superoxide dismutase. Plant Physiol
1996;111:117781.
McKersie BD, Murnaghan J, Bowley SR. Manipulating freezing tolerance in transgenic
plants. Acta Physiol Plant 1997;19:48595.
Miranda JA, Avonce N, Surez R, Thevelein JM, Dijck PV, Iturriaga G. A bifunctional TPSTPP enzyme from yeast confers tolerance to multiple and extreme abiotic-stress
conditions in transgenic Arabidopsis. Planta 2007;226(6):141121.
Mohammadi M, Taleei A, Zeinali H, Naghavi MR, Ceccarelli S, Grando Baum M. QTL
analysis for phenologic traits in doubled haploid population of barley. Int J Agric
Biol 2005;7(5):8203.
Mohanty A, Kathuria H, Ferjani A, Sakamoto A, Mohanty P, Murata N, et al. Transgenics
of an elite indica rice variety Pusa Basmati 1 harbouring the codA gene are highly
tolerant to salt stress. Theor Appl Genet 2002;106:517.
Noaman MM, El Sayad AA, Asaad FA, El Sherbini AM, El Bawab AO, El Moselhi MA, et al.
Registration of Giza 126 barley. Crop Sci 1995;35(6):1710.
Noaman MM, Ahmed IA, El-Sayed AA, Abo-El-Enin RA, El-Gamal AS, El-Sherbiny AM, et al.
Registration of Giza 2000 drought-tolerant six-rowed barley for rainfed and new
reclaimed areas in Egypt. Crop Sci 2007;47:440.
Obert DE, Evans CP, Wesenberg DM, Windes JM, Erickson CA, Jackson EW, et al.
Registration of Lenetah spring barley. J Plant Registr 2008;2(2):857.
Oh SJ, Jeong JS, Kim EH, Yi NR, Yi SI, Jang IC, et al. Matrix attachment region from
the chicken lysozyme locus reduces variability in transgene expression and
confers copy number-dependence in transgenic rice plants. Plant Cell Rep
2005;4:14554.
Oh SJ, Kim YS, Kwon C, Park HK, Jeong JS, Kim JK. Overexpression of the transcription
factor AP37 in rice improves grain yield under drought conditions. Plant Physiol
2009;2:191200. doi:10.1104/pp.109.137554.
Owttrim GW. RNA helicases and abiotic stress. Nucleic Acids Res 2006;34(11):322030.
Park BJ, Liu Z, Kanno A, Kameya T. Increased tolerance to salt and water decit stress in
transgenic lettuce (Lactuca sativa L.) by constitutive expression of LEA. Plant
Growth Regul 2005a;45:16571.
Park BJ, Liu Z, Kanno A, Kameya T. Genetic improvement of Chinese cabbage for salt and
drought tolerance by constitutive expression of a B. napus LEA gene. Plant Sci 2005b;169:
5538.
Pastori GM, Foyer CH. Common components, networks and pathways of cross tolerance
to stress. The central role of redox and abscisic acid-mediated controls. Plant
Physiol 2002;129:4608.
Pellegrineschi A, Reynolds M, Pacheco M, Brito RM, Almeraya R, Yamaguchi-Shinozaki K, et al.
Stress-induced expression in wheat of the Arabidopsis thaliana DREB1A gene delays water
stress symptoms under greenhouse conditions. Genome 2004;47:493500.
Perl A, Perl-Treves R, Galili S, Aviv D, Shalgi E, Malkin S, et al. Enhanced oxidative-stress
defense in transgenic potato overexpressing tomato Cu, Zn superoxide dismutase.
Theor Appl Genet 1993;85:56876.
Podlich DW, Winkler CR, Cooper M. Mapping as you go: an effective approach for
marker-assisted selection of complex traits. Crop Sci 2004;44:156071.

Pratt RC, Casey MA. Registration of maize germplasm line Oh605. Crop Sci 2006;46:10045.
Qin F, Kakimoto M, Sakuma Y, Maruyama K, Osakabe Y, Tran LSP, et al. Regulation and
functional analysis of ZmDREB2A in response to drought and heat stress in Zea mays L.
Plant J 2007;50:5469.
Quan R, Shang M, Zhang H, Zhao Y, Zhang J. Engineering of enhanced glycinebetaine
synthesis improves drought tolerance in maize. Plant Biotechnol J 2004;2:47786.
Quarrie SA, Gulli M, Calestani C, Steed A, Marmiroli N. Location of a gene regulating
drought-induced abscisic acid production on the long arm of chromosome 5A of
wheat. Theor Appl Genet 1994;89:794800.
Quick JS, Stromberger JA, Clayshulte S, Clifford B, Johnson JJ, Peairs FB, et al. Registration
of Prairie Red wheat. Crop Sci 2001;41:13623.
Rajaram S. Prospects and promise of wheat breeding in the 21st century. Euphytica 2001;119:
3-15.
Rajaram S. Role of conventional plant breeding and biotechnology in future wheat
production. Turk J Agric Forest 2005;29:10511.
Rami JF, Dufour P, Trouche G, Fliedel G, Mestres C, Davrieux F, et al. Quantitative trait
loci for grain quality, productivity, morphological and agronomical traits in
sorghum (Sorghum bicolor L. Moench). Theor Appl Genet 1998;97:60516.
Rassaa N, Salahb HBH, Latiri K. Thermal responses of durum wheat Triticum durum to
early water stress; consequence on leaf and ower development. Plant Biol Pathol
2008;331(5):36371.
Reddy LJ, Nigam SN, Rao RCN, Reddy NS. Registration of ICGV 87354 peanut germplasm
with drought tolerance and rust resistance. Crop Sci 2001;41:2745.
Reddy AR, Chaitanya KV, Vivekanandan M. Drought-induced responses of photosynthesis and
antioxidant metabolism in higher plants. J Plant Physiol 2004;161:1189202.
Ribaut JM, Ragot M. Marker-assisted selection to improve drought adaptation in maize: the
backcross approach, perspectives, limitations, and alternatives. J Exp Bot 2006;58:
35160.
Robin S, Pathan MS, Courtois B, Latte R, Carandang S, Lanceras S, et al. Mapping
osmotic adjustment in an advanced back-cross inbred population of rice. Theor
Appl Genet 2003;107(7):128896.
Romero C, Belles JM, Vaya JL, Serrano R, Culianez-Macia FA. Expression of the yeast
trehalose-6-phosphate synthase gene in transgenic tobacco plants: pleiotropic
phenotypes include drought tolerance. Planta 1997;201:2937.
Ronde JAD, Cress WA, Krugerd GHJ, Strasserd RJ, Van Staden J. Photosynthetic response
of transgenic soybean plants, containing an Arabidopsis P5CR gene, during heat and
drought stress. J Plant Physiol 2004;161:121124.
Salvi S, Tuberosa R. To clone or not to clone plant QTLs: present and future challenges.
Trends Plant Sci 2005;10:297304.
Sanchez AC, Subudhi PK, Rosenow DT, Nguyen HT. Mapping QTLs associated with drought
resistance in sorghum (Sorghum bicolor L. Moench). Plant Mol Biol 2002;48:71326.
Saranga Y, Menz M, Jiang CX, Wright RJ, Yakir D, Paterson AH. Genomic dissection of
genotype environment interactions conferring adaptation of cotton to arid
conditions. Genome Res 2001;11:198895.
Sari-Gorla M, Krajewski P, Di Fonzo N, Villa M, Frova C. Genetic analysis of drought
tolerance in maize by molecular markers. II. Plant height and owering. Theor Appl
Genet 1999;99:28995.
Seki M, Narusaka M, Ishida J, Nanjo T, Fujita M, Oono Y, et al. Monitoring the expression
proles of 7000 Arabidopsis genes under drought, cold, and high-salinity stresses
using a full-length cDNA microarray. Plant J 2002;31:27992.
Serraj R, Sinclair TR. Osmolyte accumulation: can it really increase crop yield under
drought conditions? Plant Cell Environ 2002;25:33341.
Serraj R, Krishnamurthy L, Kashiwagi J, Kumar J, Chandra S, Crouch JH. Variation in root
traits of chickpea (Cicer arietinum L.) grown under terminal drought. Field Crops
Res 2004;88:11527.
Serraj R, Hash CT, Rizvi MHS, Sharma A, Yadav RS, Bidinger FR. Recent advances in markerassisted selection for drought tolerance in pearl millet. Plant Prod Sci 2005;8(3):3347.
Shen YG, Du BX, Zhang WK, Zhang JS, Chen SY. AhCMO, regulated by stresses in Atriplex
hortensis, can improve drought tolerance in transgenic tobacco. Theor Appl Genet
2002;105:81521.
Shinozaki K, Yamaguchi-Shinozaki K. Molecular responses to dehydration and low
temperature: differences and cross-talk between two stress signaling pathways.
Curr Opin Plant Biol 2000;3:21723.
Singh KB, Omar M, Saxena MC, Johansen C. Registration of FLIP 87-59C, a droughttolerant chickpea germplasm line. Crop Sci 1996;36(2):12.
Singh SP, Teran H, Gutierrez JA. Registration of SEA 5 and SEA 13 drought tolerant dry
bean germplasm. Crop Sci 2001;41:2767.
Singh SP, Teran H, Lema M, Schwartz HF, Miklas PN. Registration of white mold
resistant dry bean germplasm line A 195. J Plant Registr 2007;1:623.
Sivamani E, Bahieldin A, Wraith JM. Improved biomass productivity and water use
efciency under water decit conditions in transgenic wheat constitutively
expressing the barley HVA1 gene. Plant Sci 2000;155:19.
Skovmand B, Reynolds MP, DeLacy IH. Searching genetic resources for physiological
traits with potential for increasing yield. In: Reynolds MP, Ortiz-Monasterio I,
McNab A, editors. Application of Physiology in Wheat Breeding; 2001. p. 1728.
Steele K. Novel upland rice variety bred using marker-assisted selection and clientoriented breeding released in Jharkhand. India: Bangor University; 2009.
Steele KA, Price AH, Shashidar HE, Witcombe JR. Marker-assisted selection to introgress
rice QTLs controlling root traits into an Indian upland rice variety. Theor Appl Genet
2006;112:20821.
Steele KA, Virk DS, Kumar R, Prasad SC, Witcombe JR. Field evaluation of upland rice
lines selected for QTLs controlling root traits. Field Crops Res 2007;101:1806.
Sunkar R, Kapoor A, Zhu JK. Post transcriptional induction of two Cu/Zn superoxide
dismutase genes in Arabidopsis is mediated by down regulation of miR398 and
important for oxidative stress tolerance. Plant Cell 2006;18:205165.
183
Talame V, Sanguineti MC, Chiapparino E, Bahri H, Ben Salem M, Forster BP, et al.
Identication of Hordeum spontaneum QTL alleles improving eld performance of
barley grown under rainfed conditions. Ann Appl Bot 2004;144:30920.
Tanner NK, Cordin O, Banroques J, Doere M, Linder P. The Q Motif: a newly identied motif
in DEAD box helicases may regulate ATP binding and hydrolysis. Mol Cell 2003;11:
12738.
Taramino G, Tarchini R, Ferrario S, Lee M, Pe ME. Characterization and mapping of simple
sequence repeats (SSRs) in Sorghum bicolor. Theor Appl Genet 1997;95:6672.
Teulat B, Monneveux P, Wery J, Borris C, Souyris I, Charrier A, et al. Relationships
between relative water content and growth parameters in barley: a QTL study. New
Phytol 1997;137:99-107.
Thi Lang N, Chi Buu B. Fine mapping for drought tolerance in rice (Oryza sativa L.).
Omonrice 2008;16:9-15.
Till BJ, Comai L, Henikoff S. Tillering and ecotillering for crop improvement. In:
Varshney RK, Tuberosa R, editors. Genomics-assisted crop improvement: genomics
approaches and platforms, vol. 1. ; 2007. p. 33350.
Tondelli A, Francia E, Barabaschi D, Aprile A, Skinner JS, Stockinger EJ, et al. Mapping
regulatory genes as candidates for cold and drought stress tolerance in barley.
Theor Appl Genet 2006;112:44554.
Tuberosa R, Salvi S. Genomics approaches to improve drought tolerance in crops.
Trends Plant Sci 2006;11:40512.
Valkoun JJ. Wheat pre-breeding using wild progenitors. Euphytica 2001;119:1723.
Vashisht AA, Tuteja N. Stress responsive DEAD-box helicases: a new pathway to
engineer plant stress tolerance. J Photochem Photobiol B Biol 2006;84:15060.
Vendruscolo ECG, Schuster I, Pileggi M, Scapim CA, Molinari HBC, Marur CJ, et al. Stressinduced synthesis of proline confers tolerance to water decit in transgenic wheat.
J Plant Physiol 2007;164:136776.
Vienne D, Leonardi A, Damerval C, Zivy M. Genetics of proteome variation for QTL
characterization: application to drought stress responses in maize. J Exp Bot 1999;50:
3039.
Villareal RL, Mujeeb-Kazi A, Rajaram S, Toro ED. Morphological variability in some
synthetic hexaploid wheats derived from Triticum turgidum T. tauschii. J Genet
Breed 1994;48:7-16.
Vinh NT, Paterson AH. Genome mapping and its implication for stress resistance in
plants. In: Ashraf M, Harris PJC, editors. Abiotic stresses: plant resistance through
breeding and molecular approaches; 2005.
Vinocur B, Altman A. Recent advances in engineering plant tolerance to abiotic stress:
achievements and limitations. Curr Opin Biotechnol 2005;16:12332.
Wang YJ, Hao YJ, Zhang ZG, Chen T, Zhang JS, Chen SY. Isolation of trehalose-6phosphate phosphatase gene from tobacco and its functional analysis in yeast cells.
J Plant Physiol 2005;162:21523.
Wu R, Garg A. Engineering rice plants with trehalose-producing genes improves
tolerance to drought, salt, and low temperature. ISB News Report; 2003.
Xiao B, Huang Y, Tang N, Xiong L. Overexpression of LEA gene in rice improves drought
resistance under eld conditions. Theor Appl Genet 2007;115:3546.
Xinglai P, Sangang X, Qiannying P, Yinhong S. Registration of Jinmai 50 wheat. Crop Sci
2006;46:9835.
Xiong L, Lee H, Ishitani M, Zhu JK. Regulation of osmotic stress-responsive gene
expression by the LOS6/ABA1 locus in Arabidopsis. J Biol Chem 2002;277:858896.
Xu GW, Magill CW, Shertz KF, Hart GE. A RFLP linkage map of Sorghum bicolor (L.).
Moench. Theor Appl Genet 1994;89:13945.
Xu D, Duan X, Wang B, Hong B, Ho THD, Wu R. Expression of a late embryogenesis abundant
protein gene, HVA1, from barley confers tolerance to water decit and salt stress in
transgenic rice. Plant Physiol 1996;110:24957.
Yadav RS, Hash CT, Bidinger FR, Devos KM, Howarth CJ. Genomic regions associated
with grain yield and aspects of post-owering drought tolerance in pearl millet
across stress environments and testers background. Euphytica 2004;136:26577.
Yamada M, Morishita H, Urano K, Shiozaki N, Yamaguchi-Shinozaki K, Shinozaki K, et al.
Effects of free proline accumulation in petunias under drought stress. J Exp Bot
2005;56:197581.
Yang WJ, Rich PJ, Axtell JD, Wood KV, Bonham CC, Ejeta G, et al. Genotypic variation for
glycinebetaine in sorghum. Crop Sci 2003;43:1629.
Yan-Ying QU, Ping MU, Xue-Qin L, Yu-Xiu T, Feng W, Hong-Liang Z, et al. QTL mapping
and correlations between leaf water potential and drought resistance in rice under
upland and lowland environments. Acta Agron Sin 2008;34(2):198206.
Ye XD, Wu XL, Zhao H, Frehner M, Nsberger J, Potrykus I, et al. Altered fructan accumulation
in transgenic Lolium multiorum plants expressing a Bacillus subtilis sacB gene. Plant Cell
Rep 2001;20:20512.
Zaharieva M, Gaulin E, Havaux M, Acevedo E, Monneveux P. Drought and heat responses in
the wild wheat relative Aegilops geniculata Roth: potential interest for wheat
improvement. Crop Sci 2001;41:13219.
Zhang J, Zheng HG, Aarti A, Pantuwan G, Nguyen TT, Tripathy JN, et al. Locating genomic
regions associated with components of drought resistance in rice: comparative
mapping within and across species. Theor Appl Genet 2001;103:1929.
Zhang GH, Su Q, An LJ, Wu S. Characterization and expression of a vacuolar Na+/H+
antiporter gene from the monocot halophyte Aeluropus littoralis. Plant Physiol
Biochem 2008;46:11726.
Zhao J. QTLs for oil content and their relationships to other agronomic traits in an
European Chinese oilseed rape population. Germany: Diss. Grorg-Agust University of Goettingen; 2002.
Zhao J, Ren W, Zhi D, Wang L, Xia G. Arabidopsis DREB1A/CBF3 bestowed transgenic tall
fescue increased tolerance to drought stress. Plant Cell Rep 2007;26:15218.
Zhou W, Li Y, Zhao BC, Ge RC, Shen YZ, Wang G, Huang ZJ. Over-expression of
TaSTRG gene improves salt and drought tolerance in rice. J Plant Physiol 2009;15
(166):166070.

Inducing DT in Plants Review

Enviado por

Dados do documento

Título original

Direitos autorais

Formatos disponíveis

Compartilhar este documento

Compartilhar ou incorporar documento

Opções de compartilhamento

Você considera este documento útil?

Este conteúdo é inapropriado?

Direitos autorais:

Formatos disponíveis

Inducing DT in Plants Review

Enviado por

Direitos autorais:

Formatos disponíveis

Biotechnology Advances 28 (2010) 169183

Contents lists available at ScienceDirect

Research review paper

Inducing drought tolerance in plants: Recent advances

E-mail address: ashrafbot@yahoo.com.

Breeding for any desired trait undoubtedly requires a signicant amount

M. Ashraf / Biotechnology Advances 28 (2010) 169183

genotypes that excel in the target environment (Bnziger et al., 2004).

collaboration with the International Institute of Tropical Agriculture

M. Ashraf / Biotechnology Advances 28 (2010) 169183

In barley, QTL involved in some key growth and water relation

M. Ashraf / Biotechnology Advances 28 (2010) 169183

Peanut (Arachis hypogaea

Plant Materials Identication

Reddy et al. (2001)

Derived from a cross between

Developed by crossing ILC3843 with

First breeder seed was produced from

Both lines were originated after

Giza 121, Line

Singh et al. (2001)

International Center for Tropical

Singh et al. (2001)

Centro Internacional de Agricultura

Singh et al. (2007)

Eastern Agricultural Research

Brick et al. (2008)

Bergman et al. (2001)

Singh et al. (1996)

Colorado Agricultural Experiment

Quick et al. (2001)

Wheat Breeding Innovation Group

Xinglai et al. (2006)

USDA-ARS Forage and Range Research

Jenson et al. (2007)

Kindiger et al. (2006)

Chen et al. (2007)

Obert et al. (2008)

Noaman et al. (1995)

Barley Research Department, Agricultural

Noaman et al. (2007)

Sakha Research Station Northern

Noaman et al. (2007)

M. Ashraf / Biotechnology Advances 28 (2010) 169183

Plant selections within superior rows

West Asia and North Africa (WANA)

Noaman et al. (2007)

International Center for Agricultural

Noaman et al. (1995)

Derived from crossing I92125 with

involved in drought resistance. Similarly, 14 QTL related to osmotic

Juskiw et al. (2009)

International Institute of Tropical

Badu-Apraku et al. (2006)

North Dakota Agricultural Experiment

Carena and Wanner (2009)

International Institute of Tropical

Badu-Apraku and Yallou (2009)

M. Ashraf / Biotechnology Advances 28 (2010) 169183

resulted in pyramiding all ve segments. It is imperative to note that

terminal drought conditions (Yadav et al., 2004). A mapping population

Rice (Oryza sativa L.)

QTL9 (on chromosome 9)