© 2001 Nature Publishing Group http://biotech.nature.
com
Improving rice yields—ironing out the details
Engineering crops to release more iron-solubilizing chelators may
increase their yield in alkaline soils.
Mary Lou Guerinot
© 2001 Nature Publishing Group http://biotech.nature.com
Iron is one of three nutrients (nitrogen and
phosphorus are the others) that most com-
monly limit plant growth1. Unlike other
nutrient limitations, however, iron deficiency
cannot be remedied through the use of fertil-
izers because the problem is not one of abun-
dance, but rather one of solubility. In this
issue, Takahashi et al.2 engineer rice to release
increased levels of deoxymugineic acid, an
iron-solubilizing chelator. The resultant
plants exhibit enhanced growth in soils with
low iron availability. Because half the world’s
population eat rice daily and it forms a sub-
stantial part of their diet, this dramatic
increase in yield bodes well for future provi-
sion of this important food staple.
What is it about iron that makes it so hard
for plants to acquire? Although iron is the
fourth most abundant element in the earth’s
crust, it is largely unavailable in alkaline
(pH >7) soils. In well-aerated soils with high
pH, the concentrations of Fe2+ and Fe3+ in soil
solution are extremely low, orders of magni-
tude lower than that required by plants for
optimal growth. Alkaline soils are very com-
mon in semiarid and arid regions, which
have been estimated to account for 30% of
the world’s arable soils.
Faced with a deficit of iron, plants have
two main strategies for making this essential
nutrient more available (Fig. 1; ref. 1). All
plants, except the grasses, release protons into
the soil to lower the pH, induce the expres-
sion of a Fe(III) chelate reductase to reduce
iron to the more soluble Fe(II) form3 and
induce an Fe(II) transport system to take in
the reduced iron4. Collectively, this is referred
to as a strategy I response.
Grasses, which include many of the world’s
main food crops such as wheat, rice, and
maize, release low-molecular-weight com-
pounds called phytosiderophores from their
roots in response to iron deficiency. These
phytosiderophores solubilize iron by binding
to Fe(III). The Fe(III)–phytosiderophore com-
plex is then taken up by the plant via a specif-
ic phytosiderophore receptor5. This response
is designated strategy II.
Mary Lou Guerinot is professor,
Department of Biological Sciences,
Dartmouth College, Hanover, NH 03755
(Mary.Lou.Guerinot@dartmouth.edu)
http://biotech.nature.com
Not all plants are proficient at mounting
iron-deficiency responses. For strategy II
plants, there is a strong, positive correlation
between the amount of phytosiderophores
released and the resistance of plants to iron
deficiency1. In the present paper, Takahashi et
al. hypothesized that rice is much more sus-
ceptible to iron deficiency than other grasses
because it releases low amounts of phy-
tosiderophores. The biosynthetic pathway
that leads to the formation of the rice phy-
tosiderophore, deoxymugineic acid, has been
largely elucidated, providing several targets
for increasing the amounts of phy-
tosiderophore produced.
Takahashi et al. used Agrobacterium-
mediated transformation to introduce into
rice two barley genes6 encoding nico-
tianamine aminotransferase (NAAT), the
enzyme that catalyzes the second step in the
biosynthesis of deoxymugineic acid. Under
iron deficiency, transgenic rice expressing the
two barley NAAT genes had higher levels of
NAAT activity and released larger amounts of
phytosiderophores than nontransformants.
This increased release did correlate, as pre-
dicted, with improved growth of the trans-
genic plants in alkaline soils. Remarkably, the
transgenic rice plants yielded fourfold as
Strategy I
(pea, tomato, arabidopsis)
ATP
H+
ADP
NADH FE (III)-chelate
FRO2
NAD+ FE (II)
FE (II) FE (II)
IRT1
Inside Outside
PM
Figure 1. Iron uptake by higher plants. Iron-efficient plants are able to acquire iron from soils of low
iron availability using one of two strategies. Strategy I plants reduce Fe(III) to Fe(II), which is then
transported across the plasma membrane. Strategy II plants release Fe(III) chelating siderophores
and then transport Fe(III)–siderophore complexes across the plasma membrane. Ligand exchange
and degredation are alternative fates for the Fe (III) complex. FRO2, ferric reductse-oxidase 2; IRT1,
iron-regulated transporter 1; YS1, yellow stripe 1.
• MAY 2001 • VOLUME 19 • nature biotechnology
NEWS AND VIEWS
much grain as control plants. But did the
grain itself contain more iron? This question
was left unanswered, so we cannot yet con-
clude that increased phytosiderophore
release leads to the partitioning of more iron
into the seed.
Engineering other steps in the phy-
tosiderophore biosynthetic pathway could
further increase phytosiderophore release.
The discrepancy between the high levels of
NAAT activity and the rather modest
increase in deoxymugineic acid release sug-
gests that NAAT may be limited for its sub-
strate, nicotianamine. The cloning of genes
encoding nicotianamine synthase, the
enzyme that condenses three molecules of
S-adenosyl methionine into nico-
tianamine7–9, offers the opportunity to
manipulate the synthesis of nicotianamine.
Nicotianamine itself is thought to play a role
in iron metabolism in both strategy I and
strategy II plants, possibly serving as the iron
chelator that transports iron in the phloem10.
Thus, another question that will have to be
answered is whether diverting more nico-
tianamine into the production of phy-
tosiderophores will have any effects on iron
distribution in the plant itself. This is espe-
cially important because iron is transported
into the developing seed through the
phloem. The repercussions of diverting more
S-adenosyl methionine into the production
of nicotianamine will also have to be investi-
gated. S-adenosyl methionine is the primary
donor of methyl groups in many metabolic
processes and is the immediate precursor of
the plant hormone ethylene.
Whereas increasing phytosiderophore
production looks promising for improving
Strategy II
(barley, maize, rice)
Siderophore
FE (III)-siderophore
complex
YS1
Degradation Ligand
exchange
© Bob Crimi
Inside Outside
PM
417

NEWS AND VIEWS
the growth of strategy II plants in soils with
low iron availability, what about strategy I
plants? It is possible that increased synthe-
sis of nicotianamine might confer some
benefit, but more likely gains may come
from increasing the rate-limiting step of
Fe(III) chelate reduction. And interestingly,
strategy I plants may also benefit from the
release of chelators. Plants engineered to
release more citrate into the rhizosphere
© 2001 Nature Publishing Group http://biotech.nature.com
grow better than nonengineered plants in
soils of low iron availability (Luis Herrera-
Estrella, personal communication).
Presumably, citrate forms complexes with
Fe(III) that can then be reduced at the root
surface for transport across the plasma
membrane.
In addition to improvements in the
uptake of iron from the soil, attention is
being paid to increasing the storage forms
of iron. Rice has been engineered to have
higher levels of the iron storage protein fer-
ritin in the grain, but results conflict as to
the effect of such overexpression on the
iron content of the grain11,12. And none of
the rice plants, including those described by
Takahashi et al., has been tested in the field.
The idea of fortifying foods with iron is
not new. Iron-fortified infant formula and
iron-enriched flours are widely available.
But by fortifying food crops before harvest,
we stand the best chance of ensuring that
such iron-fortified foods get to the people
who most need them. The engineering of
rice endosperm to produce more vitamin A
by Ingo Potrykus, Peter Beyer, and their co-
workers13 is the gold standard for preharvest
fortification of rice to increase its nutritional
value. The goal of producing rice with both
more vitamin A and more iron in its grain is
certainly within reach. Such rice would pro-
vide a solution for two of the three major
nutrient deficiency problems facing the
world today.
Sixty to eighty percent of the world’s pop-
ulation may be iron deficient14. Iron-
deficiency anemia is responsible for an esti-
mated 20% of maternal deaths in Africa and
Asia, and can lead to irreversible
physical/mental retardation, reduced resis-
tance to infection, and reduced work perfor-
mance. The approach described by
Takahashi et al. has a dramatic effect on
plant yield. We now eagerly await confirma-
tion that it also bolsters iron content.
1. Marschner, H. Mineral nutrition of higher plants.
(Academic Press, Boston; 1995).
2. Takahashi, M. et al. Nat. Biotechnol. 19, 466–469
(2001).
3. Robinson, N.J. et al. Nature 397, 694–697 (1999).
4. Eide, D. et al. Proc. Natl. Acad. Sci. USA 93,
5624–5628 (1996).
5. Curie, C. et al. Nature 409, 346–349 (2001).
6. Takahashi, M. et al. Plant Physiol. 121, 947–956
(1999).
418 nature biotechnology
© 2001 Nature Publishing Group http://biotech.nature.com
7. Higuchi, K. et al. Plant Physiol. 119, 471–479 (1999).
(1999). 11. Goto, F. et al. Nat. Biotechnol. 17, 282–286
8. Herbik, A. et al. Eur. J. Biochem. 265, 231–239 (1999).
(1999). 12. Drakakaki, G. et al. Transgenic Res. 9, 445–452
9. Ling, H. Q. et al. Proc. Natl. Acad. Sci. USA 96, (2000).
7098–7103 (1999). 13. Ye, X. et al. Science 287, 303–305 (2000).
10. von Wiren, N. et al. Plant Physiol. 119, 1107–1114 14. http://www.who.int/nut/ida. WHO (2001).
Searching for array standards in Rockville
A recent US NIH meeting produced a draft pilot protocol to facilitate
microarray data sharing.
Robert A. Star and Rebekah S. Rasooly
Twenty-three research groups
funded by microarray initiatives at
the National Institute on Drug
Abuse (NIDA)1 and the National
Institute of Diabetes and Digestive
and Kidney Diseases (NIDDK)2 of
the US National Institutes of
Health (NIH; Rockville, MD)
recently took part in a meeting* to
discuss current logistical chal-
lenges in microarray research and
formulate a strategy for facilitat-
ing progress in the field. After
extensive discussion, a plan was
proposed incorporating the estab-
lishment of effective communica- Figure 1. Brainstorming in the NIH Neuroscience Center in
tion tools, use of a standard subset Rockville, MD. Twenty three US scientists from the NIDA
of several hundred genes, adop- and NIDDKA met at the NIH earlier this year to hammer out
protocols for facilitating array data sharing and analysis.
tion of a single gene-naming pro-
tocol, design of a set of standard
quality controls, and the development of a Although the belief that “more data are bet-
control pool of RNA, together with the req- ter” is common, biologists are experiencing
uisite bioinformatics tools. The US difficulty in keeping up with the latest
National Center for Biotechnology advances, analyzing data, and comparing
Information (NCBI; Rockville, MD) Gene data obtained using competing technologies
Expression Omnibus (GEO) was suggested or at other laboratories. In addition, the
as an ideal repository for microarray data. field lacks uniform standards to describe
We believe that such an arrangement could simple (e.g., gene names) and complex (e.g.,
serve as a model for other national/interna- experimental details, analysis methodology)
tional microarray data curation/storage parameters.
efforts. The Rockville meeting was convened to
Biologists attempting microarray analysis discuss these issues and find a consensus for
of gene expression currently face several coordinating research. During the first day,
challenges, not least the rapid evolution of participants discussed recent advances in
array technology, methodology, statistical experimental design, methodology, and data
and bioinformatics analysis tools, and the analysis. Several speakers described method-
absence of widely accepted quality controls. ological and analytical approaches for
reducing and measuring experimental error.
Peter Munson from NIH’s Center for
Robert A. Star is senior scientific advisor and Information Technology discussed the
chief, renal diagnostics and therapeutics, importance of replicates in experimental
Division of Kidney, Urologic, and Hematologic design and the use of simple statistical tools
Diseases, NIDDK/NIH, and Rebekah S. to both assess the quality of data and distin-
Rasooly is program director, cell biology & guish outliers from the intrinsic error of the
genetics at the Division of Neuroscience & experiment. Michael Miles from the Gallo
Behavioral Research, National Institute on Center at the University of California in San
Drug Abuse/NIH, Bethesda, MD 20892-9555 Francisco outlined normalization approach-
(rrasooly@nida.nih.gov). es, including statistical methods such as pin-
• VOLUME 19 • MAY 2001 • http://biotech.nature.com