Iron Chlorosis
PVV Prasad and M Djanaguiraman, Kansas State University, Manhattan, KS, USA
Ó 2017 Elsevier Ltd. All rights reserved.
This article is a revision of the previous edition article by P.V.V. Prasad, volume 2, pp. 649–656, Ó 2003, Elsevier Ltd.
Nomenclature
Adsorption Accumulation of ions on solid surface caused
by ion exchange or other reactions.
C3 plants Plants in which the first product of carbon
dioxide fixation (photosynthetic pathway) is a three-
carbon compound.
Chelate Complex organic molecules that can combine with
ions to supply nutrients to plants at slow and steady rates.
Chlorophyll The green pigment in chloroplasts that
absorbs radiant energy and transforms it into chemical
energy.
Chloroplast A membrane-bound cell organelle which
contains photosynthetic enzymes and structures.
Chlorosis Yellowing of leaves due to lack of chlorophyll.
Dicotyledons A class of angiosperms characterized by two
cotyledons, and usually with net (reticulate) venation on
the leaves.
DNA Dioxyribose nucleic acid.
Ferredoxin An electron transferring, Fe-containing protein
that functions in photosynthesis.
Introduction
Iron (Fe) is an essential micronutrient belonging to group VIII in
the periodic table. Iron is the fourth most abundant element and
second most abundant metal in the Earth’s crust constituting
about 5% of the lithosphere; it occurs in both ferrous (Fe2þ)
and ferric (Fe3þ) forms. Most Fe in the earth’s crust is in the
form of ferromagnesium silicate. In soils, the predominant
iron oxides are hematite, goethite, lepidocrocite, magnetite,
and ferrihydrite. Iron is also a structural component of silicates
and other primary or secondary minerals (e.g., pyroxenes,
amphiboles, pyrite, and siderite). The concentration of Fe in
agricultural soils ranges from 7000 to 500 000 parts per million
(mean of 38 000 ppm or 3.8%). Although the concentration of
Fe is high in soil, it is classified as a micronutrient because the
requirement by plants is low. Although most of the Fe on the
earth crust is in the form of Fe3þ, the Fe2þ form is physiologi-
cally more significant for plants. This form is relatively soluble,
but is readily oxidized to Fe3þ, which forms precipitates. The
Fe3þ is insoluble in neutral and alkaline pH (>7) and calcareous
soils. Iron deficiency in plants generally occurs not because of
low Fe in soil, but because of various soil, plant, and climatic
factors that affect Fe availability, uptake, or metabolic use.
Iron deficiency in plants is one of the major nutritional disorders
prevalent on calcareous and sandy soils in arid and semiarid
regions of the world. It has been estimated that about one-
third of the world’s soils are calcareous and have a high potential
for Fe deficiency. It is becoming a major problem worldwide,
causing economic yield loss in numerous crops.
246 Encyclopedia of Applied Plant Sciences, 2nd edition, Volume 1
Gene A segment of the DNA double helix that acts as
a template for the production of the specific ribonucleic
acid and governs hereditary characteristics of an organism.
Monocotyledons A class of angiosperms characterized by
one cotyledon, and usually with parallel venation.
Nitrogen fixation Process in which gaseous nitrogen is
converted into nitrogenous compounds.
Nodule Enlargements or swelling on roots which contain
bacteria that fix nitrogen.
Photosynthesis Process by which plants convert light
energy into chemical energy (carbohydrates).
Respiration Process by which plants convert carbohydrates
into energy.
Rhizobia Bacteria of the genus Rhizobium, which are
associated with roots of leguminous plants and help in
nitrogen fixation.
Translocation Movement of dissolved materials from one
plant organ or tissue to another.
Location and Function
Iron was among the first micronutrient elements to be discov-
ered to be essential for plant life. Essential nutrients are those
without which plants cannot complete their life cycle; they
are irreplaceable by other elements and directly involved in
plant metabolism. Almost all Fe in the plant system is located
in the chloroplasts, the organelles that are responsible for
photosynthesis. It is also distributed in the cytoplasm and other
cell organelles, which contain additional heme and/or iron
sulfur proteins.
In plants, Fe is involved in chlorophyll synthesis, and it is
essential for the maintenance of chloroplast structure and func-
tion, photosynthesis, respiration, and nitrogen fixation and is
involved in enzyme activation and electron transfer. Iron
acts as an enzyme activator or cofactor in chlorophyll
synthesis and activates several other enzymes including cata-
lase, peroxidase, nitrate reductases, and nitrogenase. Iron plays
an important role in protein synthesis and in a series of meta-
bolic activities in respiratory enzymes and photosynthetic reac-
tions. Iron is critical in DNA synthesis through the action of the
ribonucleotide reductase. Iron is also an active cofactor of
many enzymes that are necessary for plant hormone synthesis,
such as ethylene, lipoxygenase, 1-aminocyclopropane acid-1-
carboxylic oxidase, or abscisic acid. Iron is a constituent of all
redox systems, the best known examples are enzyme systems
(heme iron structure), including prosthetic groups such as cyto-
chromes. The best-known functions of cytochromes are elec-
tron transport and the involvement of cytochrome oxidase in
http://dx.doi.org/10.1016/B978-0-12-394807-6.00122-2

the terminal step in the respiration chain. Iron also interacts
with nonheme proteins as an iron–sulfur protein (e.g., ferre-
doxin, superoxide dismutase). Iron is an important nutrient
for legumes for nodulation and nitrogen fixation. It is required
in large amounts by both the legume hosts and rhizobia.
Failure of infecting rhizobia to obtain adequate amounts of
Fe from the plant results in arrested nodule development and
failure of the host plant to fix nitrogen in adequate amounts.
Uptake and Translocation
Although Fe is one of the most abundant elements in the
earth’s crust, its uptake and subsequent utilization by plants
depend on the efficiency of its transport across the plasma
membrane of root cells. Typical iron concentrations in plant
tissues range from 10 to 100 mM. Free iron is able to generate
toxic hydroxyl radicals in Fenton reactions, which can damage
cellular components, including DNA and proteins. Therefore,
Fe uptake, distribution, and storage are tightly regulated in
plants. If the soil has high Fe content, then plants tend to use
low-affinity Fe transport systems to take up sufficient Fe to
prevent Fe toxicity. In contrast, Fe deficiency in plants usually
results in the induction of high-affinity Fe transport systems.
Iron has low mobility in the plant because it is strongly bound
within plant cells. Iron is not transferred from older to younger
leaves; therefore, its deficiency first occurs in young leaves.
Strategies of Fe Uptake
Plant species and genotypes differ in their ability to acquire Fe
from the soil. There are two specific mechanisms, strategy I and
strategy II, for increasing the solubility of Fe in the rhizosphere
and its rate of uptake by plant roots. Both of these mechanisms
are present in apical parts of the roots and respond when Fe
deficiency occurs.
Strategy I is mostly exhibited by dicotyledons and nongra-
minaceous monocotyledons. The main mechanisms are (1)
release of protons into the soil to lower the pH, favoring forma-
tion of Fe2þ, (2) induction of Fe3þ chelate reductase activity to
reduce Fe3þ to the more soluble Fe2þ form, (3) release of chem-
icals capable of reducing Fe3þ to Fe2þ and organic acids, partic-
ularly citrate, that chelate Fe in the rhizosphere, (4) induction
of an increased Fe2þ transport capacity, and (5) physiological
adaptation, such as root hair proliferation and root transfer
cell development. Examples of crops exhibiting strategy I are
soybean (Glycine max), peanut (Arachis hypogaea), sunflower
(Helianthus spp.), pea (Pisum sativum), tomato (Lycopersicon
esculentum), and other dicotyledons.
Strategy II is adopted by graminaceous monocotyledons
and is characterized by the production and release of low
molecular weight compounds termed phytosiderophores,
from their roots in response to Fe deficiency. These phytosider-
ophores solubilize Fe by binding to Fe3þ. The Fe3þ-phytosider-
ophore complex is then taken up by the plant via a high-affinity
transport system in the plasma membrane of root cells. This
mechanism is referred as strategy II. The release of phytosider-
ophores is not affected by external pH. Examples of crops
exhibiting strategy II are rice (Oryza sativa), wheat (Triticum
Plant Nutrition j Iron Chlorosis 247
spp.), corn (Zea mays; maize), barley (Hordeum vulgare), oats
(Avena sativa), and other grasses. A strong, positive correlation
between the amount of phytosiderophores released and the
resistance of plants to Fe deficiency is observed in strategy II
plants. It is also possible that Fe chelated by microbial sidero-
phores might be acquired by grasses. However, the amounts
of microbial siderophores that are produced in the rhizosphere
are probably too low to be of major significance for plant Fe
nutrition. Approximately 500 microorganisms have been
found to produce siderophores structures. Huge numbers of
siderophores have been purified and characterized chemically.
Based on their structural features and types of ligands, there are
four main classes of bacterial siderophores (phenol catecho-
lates, hydroxamates, carboxylate, and pyoverdines) and three
classes of fungal siderophores (rhodotorulic acid, ferrichromes,
and ferrichromes). Siderophores are subject to degradation
after their addition to soil, and almost all of the studies on
plant use of microbial siderophores have relied on hydroponic
experiments in which radiolabeled siderophores were added to
nutrient solutions. Iron provided by microbial siderophores is
often found to be associated with the root tissues, which may
be explained in part by uptake of Fe by microorganisms that
are associated with the root surface.
Mechanisms of Fe Transport and Storage in Plants
Transport of Fe in the plant is complex and involves many steps.
Initially, Fe is acquired by the roots and transported symplasti-
cally to the pericycle cells then to the vascular bundle and into
the xylem stream. However, little is known about transporters
that load Fe into the xylem. In the xylem, the majority of Fe is
complexed with aliphatic hydroxy acids (such as malic or citric
acid), phenols, thiols, polysaccharides, and amino acids and
transported to the shoot. Inside the plant, Fe can move in the
apoplast. After uptake from the apoplast across the plasma
membrane of root cells, the radial transport of Fe to the xylem
takes place as a Fe2þ complex with nicotianamine, a phytosider-
ophore-like chelator with high affinity for Fe. The mechanisms
of Fe unloading of the xylem are not clearly understood. This
process includes parenchyma cells and/or passive diffusion in
the apoplast as a result of transpirational water flows. The sym-
plastic pathways include transporters that are localized in cells
within the vascular cylinders. Changes in the morphology of
chlorotic leaves influence unloading of Fe through symplastic
or apoplastic pathways. Although Fe is relatively immobile, Fe
complexes can be transported in the phloem.
Chlorotic symptoms can be observed, even under sufficient
Fe supply, if plants lack ferric reductase oxidase (FRO2; encodes
root ferric chelate reductase) or Fe(II) transporter (IRT1; encodes
a ferrous Fe transporter). FRO2 and IRT1 genes are induced by Fe
deficiency in Arabidopsis. Their expression is dependent on the
regulator gene FRU (¼ FER-like regulator of iron uptake). The
IRT1 transporter moves Fe2þ across the plasma membrane of
root epidermal cells. IRT1 is the major Fe transporter responsible
for Fe uptake from the soil, and it also transports manganese,
zinc, and cadmium. The expression of FRO2 and IRT1 is depen-
dent on the regulator gene FRU (¼ FER-like regulator of iron
uptake), encoding a basic helix–loop–helix (bHLH)-type tran-
scription factor. This bHLH gene was identified as an iron
248 Plant Nutrition j Iron Chlorosis
regulator and named FIT1 (¼ Fe deficiency–induced transcrip-
tion factor 1). In tomato, a homologous system exists including
LeIRT1, LeFRO1, and the bHLH factor LeFER. Phytosiderophores
are chemically distinct from bacterial and fungal
siderophores and belong to the mugineic acids (MAs) class of
compounds. The biosynthesis of MAs starts from the condensa-
tion of three S-adenosyl methionine molecules to form the
precursor, nicotianamine, by the enzyme nicotianamine
synthase (NAS) which is then converted to a 30 -keto
intermediate by NA aminotransferase (NAAT), and this
intermediate is then reduced to produce deoxymugineic acid
by the action of deoxymugineic acid synthase. This
deoxymugineic acid is then released into the rhizosphere
through the MAs1 transporter. The genes of the MA
biosynthetic pathway have been cloned and characterized in
barley, rice, maize, and wheat. The expression of these genes is
upregulated by iron deficiency through transcriptional
regulation that is mediated by various combinations of cis-
acting elements and trans-acting factors. Iron deficiency
responsive elements 1 and 2 (IDE1 and IDE2, respectively)
were the first identified cis-acting elements controlling gene
expression in response to micronutrient deficiencies. The Fe3þ-
phytosiderophore uptake transporter is called yellow stripe 1
(YS1). The YS1 proteins are distantly related to the
oligopeptide transporter family of transporters, and it is
a proton-coupled symporter of Fe3þ-phytosiderophore
complexes.
Members of the ZIP transporter family are involved in Fe
transport within the plant. This family is divided into four
subgroups based on amino acid sequence similarity: subfamily
I, subfamily II, LIV-1, and gufA (gene of unknown function).
ZIP proteins transport metal ions from the cell exterior or
lumen of intracellular organelles into the cytoplasm. Arabidop-
sis is predicted to have 15 ZIP genes belonging to subfamily I,
and these ZIP genes are involved in metal transport from the
soil into root cells and across both cellular and organelle
membranes. Different ZIP transporters function in different
tissues and with different substrate affinities and specificities
to affect particular transport functions.
Iron is immobilized in plant tissues, particularly in the
leaf, making it unavailable for physiological and biochemical
functions. It has been observed that Fe is commonly immobi-
lized close to vascular systems in Fe-deficient leaves. Iron-
deficient leaves accumulate more Fe in the midrib and veins,
and concentrations of Fe in the mesophyll are lower than in
Fe-replete plants. The forms of accumulation of Fe are not
clearly known. There are indications that Fe can be stored
temporarily in plant tissues with ferritins (Fe storage proteins
located in the plastids) or other proteins that can be remobi-
lized within the plant to support critical functions, such as
photosynthesis and respiration, and cellular organelles such
as chloroplasts and mitochondria. The Fe storage protein
ferritin accumulates mainly in nongreen plastids, such as etio-
plasts or amyloplasts, while low concentrations of this protein
are found in mature chloroplasts where the photosynthetic
process is active. Iron can also be stored in apoplasmic space,
between the plasma membrane and the cell wall of plant cells,
and perhaps in the vacuoles, where low pH and high organic
acid concentrations represent optimal conditions for Fe
deposition.
Diagnosis of Fe Deficiency
Diagnosis of a nutrient deficiency is usually demonstrated by
one or more of three methods: visual observation of deficiency
symptoms, soil analysis, and plant analysis. Integration of all
three methods is essential for accurate diagnosis of Fe
deficiency.
Visual Symptoms
In most plant species, Fe deficiency is characterized by intervei-
nal chlorosis with fine reticulate patterns observed in newly
formed leaves. The dark green veins are clearly visible against
a yellow background (Figure 1) as shown in common bean.
In severe cases, the youngest leaves are completely white and
devoid of chlorophyll or develop necrotic spots, which can
look like a fungal infection, on the affected leaves. Over time,
plants which remain untreated will start to senesce, become
unsightly in appearance, and may eventually die. Iron defi-
ciency can easily be mistaken for nitrogen or magnesium defi-
ciency; however, Fe deficiency affects the young emerging
leaves, whereas nitrogen deficiency affects old leaves. This is
because Fe is relatively immobile in the phloem compared
with nitrogen or magnesium.
Soil Analysis
Soil analysis is often used to determine whether the supply of
a particular nutrient might limit plant growth. However, soil
analysis for predicting Fe deficiency in plants cannot be recom-
mended. Soil analysis to determine Fe availability is
confounded by soil environmental factors such as pH, mois-
ture content, temperature, and bicarbonate concentration,
and the genetic variability within and between plant species
with respect to Fe uptake. Several methods have been devised
to extract Fe from soil, the most commonly used being the
diethylenetriaminepentaacetic acid (DTPA) extraction method.
The critical range for Fe for this test is 2.5–4.5 ppm. If the soil
contains less than 2.5 ppm, it is considered deficient in Fe.
However, even if soils are sufficient in Fe, this does not mean
that the plant will not suffer Fe chlorosis. The uptake of Fe by
plants depends on many other factors besides extractable Fe
in the soil. These factors are discussed below.
Plant Analysis
The prediction of micronutrient deficiencies based on tissue
analysis has been successfully used for all micronutrients
except Fe. The total Fe concentration in plant tissues does
not correlate well with the occurrence of Fe chlorosis. This is
mainly because only 10–20% of the total Fe in plants is ‘phys-
iologically active’ or ‘available Fe.’ Several techniques based
on plant tissue analysis have been proposed for diagnosis of
Fe deficiency in plants. Various extractants such as water,
dilute acids (hydrochloric acid, acetic acid, oxalic acid, and
citric acid), dilute NaOH, chelating agents such as ethylene-
diaminetetraacetic acid (EDTA), DTPA, tartaric acid, and
some organic solvents including 2,20 -dipyridyl and its deriva-
tives, and o-phenanthroline have been proposed to extract the
fraction of total Fe which is metabolically active. Results

Figure 1 Typical progressive symptoms of Fe deficiency in common bean. Reproduced with permission from the American Phytopathological
Society Press from Nutrient Deficiencies and Toxicities of Plants – Digital Image Collection CD Rom, 2000. APS Press, Minnesota.
indicate that the o-phenanthroline-extractable Fe of fresh leaf
tissue is a better index of plant Fe status than total Fe content.
Another quantitative measure for diagnosing Fe deficiency is
the estimation of chlorophyll content, but this should be
used with caution as chlorophyll deficiency can be caused
by the deficiency of other nutrients (for example, N, S, Mg,
and Mn) as well.
Causes of Fe Deficiency
Various factors related to soil, climate, and plant genotype that
can contribute to Fe chlorosis are summarized below.
Soil Factors
The availability of Fe to plants largely depends on the amount
of Fe in the rocks from which the soil parent material was
derived. In addition, high pH, high clay content (<10–15%
clay), high Ca content, high carbonate content, wet conditions,
high P content, high heavy metal content, low or high temper-
atures, high light intensities, high nitrate–nitrogen content,
poor aeration, and low organic matter in soil can all contribute
to reduced Fe uptake and Fe deficiency in plants.
Soil pH
The availability and uptake of nutrients by plants are highly
dependent on pH. Iron chlorosis is common in alkaline soils
(pH greater than 7.0). The solubility of Fe is highly pH depen-
dent, and the activities of Fe3þ and Fe2þ decrease by 1000-fold
and 100-fold, respectively, for each unit increase in pH. At
acidic pH values, phosphate ions react with aluminum and
iron to form less soluble compounds. Under alkaline condi-
tions, Fe2þ is oxidized to Fe3þ, which is relatively unavailable
to plants because it precipitates as ferric oxide, which has
extremely low solubility.
Plant Nutrition j Iron Chlorosis 249
Lime Content
Iron availability is low in calcareous soils, which contain a high
concentration of lime (calcium carbonate). In the presence of
water and carbon dioxide, lime decomposes, resulting in accu-
mulation of bicarbonates and calcium, as shown in eqn. [1],
which in turn increases pH:
CaCO3 þ CO2 þ H2 O / Ca2þ þ 2HCO3  [1]
Severe Fe chlorosis occurs in soils that have 420% total CaCO3
and/or 410% active lime. Chlorosis caused by lime is often
termed lime-induced chlorosis. Studies have shown that leaves
suffering from lime-induced chlorosis often have a high
concentration of total Fe similar to or even greater than that of
green leaves, indicating a physiological inactivation of Fe in the
leaf apoplast due to the alkalization process. Furthermore, it was
observed that the severe symptoms of lime-induced chlorosis are
often correlated not only with greater total Fe concentrations in
leaves (on a dry weight basis), but also with severe inhibition of
leaf growth and chloroplast development.
Bicarbonate Content
Bicarbonates (HCO3  ) in soil and water are an important cause
of Fe chlorosis. Bicarbonate ions can be formed in calcareous
soils by the reaction of CO2 and water on calcite. Many alkaline
soils also have high bicarbonate concentration, which can
inhibit Fe uptake. Poor soil moisture, accumulation of CO2
produced by roots, and microbial respiration under high soil
moisture conditions can increase the accumulation of HCO3
in the rhizosphere up to 400–500 ppm. High HCO3 concentra-
tion inhibits root growth, thus decreasing the rate of cytokinin
export to the shoot, which is necessary for protein synthesis
and chloroplast development, resulting in chlorosis.
Organic Matter
Available Fe in soils is primarily present as organic complexes.
Organic matter in soils thus exerts a pronounced effect on Fe
250 Plant Nutrition j Iron Chlorosis
availability. The formation of soluble Fe complexes by natu-
rally occurring chelates may enhance the solubility of Fe.
However, heavy manuring in alkaline soils decreases Fe avail-
ability as it is strongly adsorbed on the surface of organic
matter; however, on decomposing, Fe is slowly supplied to
plants.
Nutrient Interactions
Iron deficiency can be induced by the interactions of Fe with
various other nutrients, for example, N, P, K, Ca, Mg, and
Zn. The form of nitrogen applied may affect the availability
of Fe. Increased uptake of NO3-N (nitrate–nitrogen) may
cause an imbalance in the cation/anion ratio, resulting in alka-
linization of the rhizosphere with a subsequent decrease in Fe
uptake. Nitrate uptake also leads to alkalization of the root
zone, which can lower Fe solubility and availability for uptake
by roots. In contrast, NH4-N (ammonical nitrogen) fertilizers
acidify the soil and can improve Fe acquisition by roots. High
P concentrations in soils decrease Fe availability to plants due
to formation of insoluble Fe phosphates. Similarly, high P
concentrations in plant tissues may also induce Fe chlorosis
due to the immobilization of Fe in the veins of the leaves.
Studies have shown that certain Fe-efficient plants are
unable to respond to Fe deficiency stress in the absence of
potassium (K). Potassium seems to play a very specific role in
plants for maximum utilization of Fe. This might be due to
its role in the production and release of protons and
phytosiderophores.
Environmental Factors
Climatic factors greatly influence the occurrence of Fe defi-
ciency in plants under field conditions, the most important
being temperature and moisture content.
Temperature
Since Fe uptake by roots and translocation from roots to shoots
is an active process, temperature influences the occurrence of Fe
deficiency. Temperature could influence the severity of Fe defi-
ciency in plants in the following ways: (1) low soil temperature
reduces root growth and metabolic activity, thus reducing Fe
uptake; (2) low soil temperature could reduce the production
of phytosiderophores, and the resultant mobilization and
uptake of soil Fe; (3) low soil temperature could increase
HCO3 levels in the soil and severity of chlorosis by increasing
the solubility of CO2 in soils; (4) high soil temperature decreases
Fe uptake by increasing microbial decomposition of phytosider-
ophores; and (5) high soil temperature could increase HCO3
levels and Fe chlorosis by stimulating microbial activity and
CO2 production.
Soil Moisture
A high soil moisture level has a strong effect on the appearance
of Fe chlorosis through its effect on plant metabolism. Many
studies have indicated that excess irrigation or prolonged wet
periods in calcareous soils result in Fe chlorosis because of
HCO3 buildup. Increased Fe chlorosis in plants subsequent
to irrigation is sometimes due to high levels of HCO3 in irriga-
tion water. In addition, high HCO3, high pH, and low Fe
content in poorly aerated soils caused by excess water destroy
many of the smaller roots and reduce the uptake capacity of
the whole root system, which may induce Fe chlorosis.
However, in waterlogged soils (especially in paddy rice
fields), Fe3þ compounds, particularly the amorphous forms,
are reduced to Fe2þ by anaerobic bacteria that use Fe3þ as an
electron acceptor during respiration (Eqn. [2]). This results in
a high solubility of Fe in flooded soils, leading to toxic levels
of Fe (450 mg kg1).
Fe(OH)3 þ 3Hþ þ e / Fe2þ þ 3H2O (2)
Plant Factors
Different species and even cultivars within the same species
vary in their susceptibility to Fe deficiency. Variation in the
ability to acquire and translocate Fe within the plant has
been observed in many species. Some species or cultivars
respond to Fe deficiency by inducing biochemical and physio-
logical responses that make Fe more available for uptake by the
roots. These plants are classified as Fe efficient. Plants that are
unable to develop such mechanisms are classified as Fe ineffi-
cient. Fe-efficient species have a greater tendency to lower the
pH of the rhizosphere. These responses enable Fe-efficient
species to acquire more Fe. Some species are also stimulated
to form more root hairs and develop cells that increase the
capacity to reduce Fe3þ in roots.
Effects of Fe Deficiency on Plants
About 9% of total leaf Fe is found in leaves as heme iron.
Nonheme iron proteins contain about 19% of the total Fe,
and they are found in ferredoxin, thylakoid complexes, mito-
chondrial complexes, aconitase, nitrite reductase, and sulfite
reductase. Most of the remainder of the Fe is found as ferritin
(35%). Thus, Fe proteins contain about 63% of the total Fe
in leaves.
Nodule Development
Severe Fe deficiency decreases root growth and elongation. In
most legumes, early nodule development after nodule initia-
tion is most sensitive to Fe deficiency. For example, in peanut,
Fe deficiency decreased the number of excisable nodules,
nodule mass, number of bacteroids and concentrations of
leghemoglobin, nitrogenase activity, and nitrogen fixing ability
(Table 1). Iron deficiency did not limit the growth or soil and
rhizosphere populations of peanut Bradyrhizobium, and there
was no effect on root infection processes or nodule initiation.
Photosynthesis and Electron Transfer
Current knowledge tends to support the hypothesis that Fe is
involved in 5-aminolevulinic acid (ALA) synthesis. The ALA,
a precursor of chlorophyll formation, is formed primarily via
a five-carbon pathway, using glutamate or a-ketoglutarate. Fe
chlorosis decreases the level of chlorophyll, membrane protein,
and other carotenoids in all plant species, e.g., in grape (Vitis
spp.; Table 2). As chlorophyll is essential for photosynthesis,
Fe deficiency leads to decreased photosynthesis. The net
 Plant Nutrition j Iron Chlorosis 251

Table 1 Effect of Fe deficiency on different aspects of nodulation in Table 3 Effect of Fe chlorosis on growth and
peanut yield of peanut

Trait Fe þFe Trait Fe þFe

Number of nodule initials (plant1) 55 52 Leaf area (cm2) 429 562

Number of nodules on tap root 11 32 Leaf dry weight (g plant1) 4.0 4.9
Nodule mass (mg plant1) 15 60 Stem dry weight (g plant1) 4.2 4.9
Leghemoglobin concentrations (nmol g1 fresh 6 86 Root dry weight (g plant1) 0.41 0.48
weight nodule) Pod dry weight (g plant1) 1340 1660
Bacteriod content (106 plant1) 2 430
Bacteriod concentration (106 fresh weight nodule) 170 7300 Treated plants received foliar application on 0.5% (w/c)
FeSO4 at 40, 60, and 90 days after sowing.
Acetylene reduction activity (pmol min1 plant1) 3 35

Adapted with permission from O’Hara, et al., 1988. New Phytol. 108, 51–57. Treated
plants received foliar application of 0.5% FeSO4 in 0.75% Tween-80 on day-10. Iron Deficiency in Field Crops
Nodules were harvested from plants on day-15 for analysis.

Iron deficiencies occur in many crops and different geograph-

Table 2 Effect of different phases of Fe deficiency on ical regions. The relative sensitivity or tolerance of crops to
photosynthetic pigments and components that can influence low soil available Fe is shown in Table 4. Iron deficiency is
photosynthesis most common in sensitive crop species grown in arid and
semiarid regions with calcareous soils. Iron deficiency is
Trait Control Mild chlorosis Severe chlorosis widely reported in soybean, peanut, dry bean, sorghum,
Chlorophyll (a þ b) 2.25 1.30 0.36 and rice. Although not common, Fe deficiency can also occur
Carotenoids 0.85 0.56 0.20 under particular conditions in corn, wheat, oat, and other
Electron transport chain 149 97 50 minor crops. Among the various field crops, the response
Photosystem I 368 351 324 of soybean to Fe deficiency has been studied in most detail.
Photosystem II 240 119 48 Iron deficiency can cause a significant reduction in yield
Rubisco activity 56 33 15 of soybean. The acreage experiencing Fe deficiency has
Proteins 49 37 23 increased significantly in the last three decades, causing
significant yield losses. A wide genetic variability exists
Adapted with permission from Bertamini, et al., 2001. Photosynthetica 39, 59–65.
among soybean varieties for susceptibility to Fe deficiency.
Roots of highly resistant (Fe-efficient) cultivars have been
reduction in photosynthesis associated with Fe deficiency shown to reduce Fe3þ to Fe2þ more actively than those of
might arise from reduction in absorbed light due to decreased highly susceptible genotypes under Fe deficiency stress.
pigments and/or reduction in porphyrin components of the Peanut is another dicotyledonous species sensitive to Fe defi-
light harvesting apparatus. Among the various processes of ciency. In peanut grown as a monocrop on calcareous soil, Fe
photosynthesis, photosystem II is more sensitive to Fe chlorosis deficiency is a major crop production constraint. However,
than photosystem I. Fe chlorosis decreases all components of when peanut is intercropped with maize, the occurrence of
the electron transport chain. Owing to the direct involvement Fe deficiency is relatively low. The mechanism associated
of Fe in protein synthesis, Fe deficiency decreases the amounts with improved Fe uptake is complex and not well under-
of many proteins, including ribulose-1,5-bisphosphate stood. Iron deficiency is also a problem in dry bean.
(rubisco), which plays an important role in the carbon cycle However, similar to soybean, there exists large genetic vari-
of C3 plants. ability among the varieties of dry bean in their susceptibility
to Fe deficiency. The most tolerant cultivars remain green all
season with normal yields, and the sensitive cultivars suffer
Growth and Yield
severe yield loss.
Iron deficiency can decrease the growth and yield of crops Sorghum releases naturally occurring chelates (phytosi-
significantly. For example, in peanuts grown in calcareous soils, derophores) in response to Fe deficiency, but the magnitude
Fe deficiency decreased leaf area and dry matter production of of phytosiderophore release is less than more tolerant crops
leaves, stems, and root, leading to yield losses of 20% (Table 3). such as corn and wheat. Iron deficiency symptoms in
Yield losses to the extent of 25% due to Fe deficiency are re- sorghum are typically expressed in young plants, but become
ported for several other crops such as cereals (rice, wheat, obvious at later growth stages resulting in significant yield
corn, and sorghum), legumes (soybean, dry bean, mung bean decrease. Fe deficiency at early stages causes uneven flower-
(Vigna spp.), and lupin (Lupinus albus)), vegetables (tomato, ing and delayed grain ripening. In severe cases, Fe deficiency
capsicum (Capsicum spp.), potato (Solanum tuberosum), and can cause a total crop failure. Rice is moderately sensitive to
spinach (Spinacia oleracea)), and several fruit crops (citrus Fe deficiency; however, the problem is relatively rare. Rice is
(Citrus spp.), grape (Vitus vinifera), pears (Pyrus communis), generally grown under flooded condition; under these condi-
and apple (Malus pumila)). Similarly, the growth and value of tions, soil Fe is reduced to the more soluble Fe2þ forms, even
commercial ornamental crops, such as rose (Rosa spp.), chry- in calcareous soils. Like sorghum, rice is also very sensitive to
santhemum (Chrysanthemum spp.), and tulips (Tulipa spp.), iron deficiency at early stages resulting in reduced yields.
are reduced by Fe deficiency. Iron deficiency in rice depends on irrigation and flooding
252 Plant Nutrition j Iron Chlorosis

Table 4 Relative sensitivity of various plants to Fe deficiency

Susceptible Moderately susceptible Relatively tolerant

Field bean (Vicia faba) Alfalfa (Medicago sativa) Barley (Hordeum vulgare)
Sorghum (Sorghum bicolor) Corn or maize (Zea mays) Millets (Pennisetum spp.)
Peanut (Arachis hypogaea) Cotton (Gossypium spp.) Oat (Avena sativa)
Soybean (Glycine max) Field pea (Pisum sativum subsp. arvense) Potato (Solanum tuberosum)
Sudangrass (Sorghum  drummondii) Forage legumes Sugar beet (Beta vulgaris subsp. vulgaris)
Upland rice (Oryza sativa) Wheat (Triticum spp.)
Dry (common) bean (Phaseolus vulgaris) Rye (Secale cereale)
Grape (Vitis spp.) Sunflower (Helianthus spp.)
Maple (Acer spp.) Amaranthus spp.
Peppermint (Mentha piperita)

practices. Maize is moderately sensitive to Fe deficiency. The Inorganic Fe Compounds

symptoms are frequent in soils with high pH and sodium
content. Iron deficiency in sodic soils suggests that the Fe
uptake mechanisms are impaired as a result of salinity stress.
Wheat and oat are tolerant to Fe deficiency, even when
grown in soils where Fe deficiency may be a problem with
other crops. Grazing or clipping is known to intensify Fe
chlorosis. The relatively tolerant nature of these crops may
be due to release of adequate phytosiderophore for Fe
uptake.
Management of Fe Deficiency
Some of the important practices adopted to alleviate Fe chlo-
rosis are soil amendments, foliar application of Fe compounds,
genotypic selection, and agronomic crop management prac-
tices. Further research is required on novel methods of applica-
tion of Fe compounds, use of adjuvants, and also analyses of Fe
compound sin various sources.
Soil Additives
Soil additives for control of Fe chlorosis are mainly categorized
into inorganic Fe salts, Fe chelates, and other organic and acid-
ifying soil amendments. A list of some inorganic and organic Fe
fertilizers is given in Table 5.
The most common inorganic source of Fe is FeSO4. Soil appli-
cation of inorganic FeSO4 can alleviate Fe chlorosis when soils
are deficient in Fe. Soil applications of inorganic Fe sources are
usually not very effective in supplying Fe for crops unless
applied in high doses, which is rarely economical for field
crops. Inorganic Fe sources, when applied to soil, are rapidly
converted into forms of Fe that are not available to plants, espe-
cially in calcareous soils.
Iron Chelates
The term ‘chelate’ refers to a chemical that forms complexes
with certain micronutrients, protecting them from becoming
unavailable. The effectiveness of Fe chelates depends princi-
pally on their reactivity. The mechanism of chelate-assisted Fe
uptake by roots may be described in four steps: (1) Fe3þ
must be released from the Fe chelate, so that the plant can
reduce it and take up Fe2þ, leaving the free chelating agent in
the soil, (2) the free chelating agent moves from the root
surface into soil by diffusion, (3) the chelating agent dissolves
the native Fe in the soil, and (4) the Fe chelate returns to root
surface by mass flow or diffusion. The Fe that reaches the plant
is the amount added minus the amount precipitated. Soil
applications of chelated compounds are more effective than
inorganic iron salts in correcting Fe chlorosis, but are too
expensive for general use, except for commercial crops of
high value.

Table 5 List of various sources of Fe fertilizer and their Fe content (%)

Fertilizer Formula Fe (%)

Inorganic
Ferrous sulfate FeSO4$7H2O 19
Ferric sulfate Fe2(SO4)3$4H2O 23
Ferrous oxide FeO 77
Ferric oxide Fe2O3 69
Ferrous ammonium sulfate FeSO4$(NH4)2$SO4$6H2O 14
Ferrous ammonium phosphate Fe(NH4)$PO4$H2O 29
Iron ammonium polyphosphate Fe(NH4)$HP2O7 22
Organic
Na-Fe ethylenediaminetetraacetate (EDTA) 5–4
Na-Fe diethylenetriaminepentaacetate (DTPA) 10
Na-Fe hydroxylethylenediaminetetraacetate (HEDA) 5–9
Na-Fe ethylenediaminedi(o-hydroxylphenol acetate) (EDDHA) 6

Slow Release Iron Fertilizers
In recent years there have been significant improvements in
Fe fertilizer formulations that include the development of
slow release Fe fertilizers and sources that are environmen-
tally friendly. The new slow release fertilizers are water insol-
uble and contain linear chain polymerized phosphates.
These phosphates are solubilized by compounds that have
affinity toward Fe such as citrates and DPTA. Thus, these
compounds can be solubilized by root exudates and make
the Fe available. There have also been improvements in
developing biodegradable, synthetic chelating agents that
are structurally similar to EDTA or EDDS (ethylenediamine-
disuccinic acid). Research is currently underway for testing
these new products in the field.
Other Soil Amendments
Organic materials such as plant residues, manures, sewage
sludge, and peat are good carriers of Fe and are effective in alle-
viating Fe chlorosis. Similarly, the application of farmyard
manure and green manure to soil has shown improvement of
Fe deficiency. One of the best ways to increase the availability
of Fe in the soil is to reduce the pH of the soil. Soil amelioration
to prevent Fe chlorosis by acidification of the entire root zone is
not practical. Therefore, part of the soil near the root zone is
acidified by the application of sulfur or other acidifying agents.
The amount of acidifying materials required varies with the
amount of CaCO3 present in the soil.
Foliar Management
Inorganic and chelated forms of Fe fertilizers are used as foliar
fertilizers for correction of Fe deficiency in crop plants. These
include FeSO4, FeEDTA, FeDTPA, FeEDDHA, Fe citrate, and
FeIDHA (iminodisuccinic acid). As soil applications of most
Fe fertilizers are generally less effective than foliar applications,
foliar application is widely used for the correction of Fe chlo-
rosis. Both inorganic and organic Fe sources are effective as
foliar sprays. The spraying of FeSO4 solution together with
some surfactant is very effective in correcting Fe chlorosis in
many crops. However, the effectiveness of Fe compounds in
overcoming Fe deficiency chlorosis is highly variable and
depends on their stability, ability to penetrate the leaf cuticle,
and their mobility/translocation following diffusion into leaf
tissue. Because of the poor translocation of applied Fe within
Table 6 Examples of Fe efficient and inefficient cultivars in important field crops
Crops Fe-efficient cultivars
Rice ARC 10372, Cauvery, TR 23, TR 25, NC 85021, IET 7972,
and IET 7973
Wheat N 85021, Aroona, Excaibur, Stiletto, Trident
Maize WF 9
Sorghum QL3, SC 630-11E, SC33-9-8-E4, 80-4124, Kanol 1, K1, K
8, K 10, K 11, CSV 15, Co 8
Soybean Bragg, Pioneer, A7, Dawson, Kiroyogradskaya4,
Veselovskaia 1, VIR 1187
Peanut ICGV 86031, ICGV 06146, 77-234, 71-58,
Mung bean UT1, CN 36, CN 60, CNM1, NM10-12-1
Field bean Santi, Px 95-183-7-1, Px-89-82-1, Px 97-58-1
Plant Nutrition j Iron Chlorosis 253
the plant, the applied Fe does not move readily from sprayed
parts to other parts of the plant. However, the use of urea
together with Fe fertilizers has been observed to increase grain
Fe concentrations. The rate of translocation of Fe from leaves
varies with crop type but is always less than 50% of applied
Fe to a given leaf or leaflet. Therefore, under field conditions,
growers must apply foliar sprays of Fe repeatedly to provide
adequate amounts to the developing canopy.
Cultivar Selection
Certain cultivars are more efficient in utilizing Fe and are less
affected by Fe deficiency. The main mechanisms involved in
Fe-efficient cultivars of dicotyledons include (1) the ability to
reduce pH due to Hþ efflux from roots; (2) enhanced root-asso-
ciated Fe3þ reduction; (3) secretion of citrate in the roots; (4)
increased root hair development; and (5) greater ability to
form chelates with Fe to improve uptake and translocation.
In monocotyledons, it is the ability of a cultivar to produce
phytosiderophores that correlate with their efficiency in
utilizing Fe. Since Fe efficiency is genetically determined, the
best long-term solution would be the development of cultivars
that are tolerant to Fe deficiency. These could easily be adopted
by farmers and would be the most efficient way to alleviate Fe
deficiency in crops. Examples of Fe-efficient cultivars of major
food crops are given in Table 6.
Biofortification of Edible Crops with Fe
Iron deficiency is also a major problem in humans. Iron defi-
ciency in humans is widespread in developing countries, where
it is estimated that 40–45% of school-age children are anemic;
approximately 50% of this anemia results from Fe deficiency.
In humans, Fe deficiency is mainly the result of plant-based
diets, which contain low levels of poorly bioavailable Fe.
Increasing bioavailable Fe in human diets can ameliorate
anemia. However, in developing countries, populations are
more dependent on plant sources, such as cereals, that contain
low Fe concentrations and high concentrations of antinutritional
compounds such as phytates which render Fe unavailable for
absorption by the gut. Thus, enhancing Fe concentrations in
edible crops can not only help to increase crop productivity
but also remedy anemia in human populations. Traditional
Fe inefficient cultivars
Pusa 33, Mahsuri, Saket 4
Durati, Yallaroi
YS 1
SC 118-15E, SC 369-3-1JB, Redland, TX 2775, TAM 428, KS 56,
80-3653, 80-3707, 80-3782, DMS 652, TNS 597, Co 21, Co 25
Throne, Hei Nong No 16, L 117
R 9227, R 8808, Dh 2000-1, TMV 2
CNM 8509B, Kamphaeng Saen 2
Parafield, Glenroy, Px 97-9-4, Px 96-83-1-1
254 Plant Nutrition j Iron Chlorosis
strategies to deliver Fe to humans are through supplementation.
An alternative solution to increase Fe concentrations in edible
crops is termed as biofortification and can be achieved by crop
management or plant breeding. Research is currently underway
to improve the uptake of Fe by plants from the soil and to
produce rice plants (and other food crops) that store greater
concentrations of Fe in bioavailable forms.
Considerable genetic variation in tissue Fe concentrations
exists in crop species, and this can be harnessed for bio-
fortification (Table 6). Varieties with greater mineral concentra-
tions in their edible portions are already available, and new
genotypes with higher mineral densities are being developed.
Screening germplasm collections has indicated a large genetic
variation in the Fe concentrations in the edible parts of staple
foods (e.g., wheat, bean, cassava, maize, rice, and yam). For
example, the Fe content of wheat cultivars varied from 25 to
56 mg kg1. The Fe content in plants and Fe concentrations in
edible parts might be increased by increasing Fe uptake from
the soil and Fe storage within the edible parts. Increasing the levels
of siderophores, chelating agents, reducing agents, enzymes, and
transporter proteins in roots could increase Fe uptake.
Engineering Fe-Efficient Crops and Increasing Fe
Content
Another approach to correct Fe deficiency in crops and humans
is to genetically modify Fe acquisition in crops by improving
their Fe efficiency. In several crops (e.g., oat, chickpea, pepper,
sunflower, and rice), the Fe efficiency trait is determined by one
or two genes and is simply inherited. In other crops (e.g.,
sorghum, clover, and soybean), Fe efficiency is a complex trait
determined by several genes acting in a quantitative manner. In
some crops (e.g., oat, dry bean, sorghum, and soybean),
breeding methods used successfully to improve Fe efficiency
of cultivars include recurrent selection and the backcross and
pedigree methods. In soybean, several different population
development strategies (single crosses, backcrosses, and three-
way crosses and four-way crosses) are used to select high-
yielding cultivars with improved Fe efficiency. Iron content is
known to vary in wild wheat (25–56 ppm), and Fe is positively
correlated with Zn content. Synthetic hexaploid wheat lines
(Triticum durum  Aegilops tauschii) have also been identified
as valuable sources of germplasm to increase the concentration
of iron. Thus it is possible to cross wild species with cultured
varieties to increase the micronutrient concentrations in grain.
An alternative way to increase the Fe concentrations in
edible parts is through a transgenic approach. It is observed
that expressing a yeast ferric reductase gene in transgenic
tobacco plants increased leaf Fe concentrations by 50%. Simi-
larly, increasing the concentration of the storage proteins phy-
toferritin also increases Fe concentrations. Transgenic rice
expressing the soybean ferritin gene was shown to have signif-
icantly greater grain Fe concentration than the untransformed
control. Cooking and processing of grain can negatively impact
Fe availability. It may be possible to introduce heat- and acid-
resistant phytases into cereal grains to withstand cooking and
degrade phytic acid in the gut during digestion, thereby
increasing Fe absorption by humans. Finally, another option
would be to introduce Fe absorption enhancers, such as
ascorbic acid, cysteine-containing peptides or hemoglobin,
into the edible plant tissues.
In strategy I plants, reduction of Fe3þ to Fe2þ is dependent
on the enzyme ferric reductase; therefore, Fe efficiency of plants
can be improved by manipulating expression of this enzyme. It
has been established that the FRO2 gene in Arabidopsis
encodes expression of ferric chelate reductase and is expressed
under Fe deficiency. Similarly, the ferric reductase gene FRE1
was identified in yeast. Genetically modified lines of tobacco
(Nicotiana tabacum) with FRE1 genes showed greater tolerance
to Fe deficiency.
Ferritin is an iron storage protein capable of binding up to
4500 Fe atoms per molecule and is a major source of Fe. Ferritin
from legumes represents an attractive target protein for
improving rice grain Fe concentration. Comparing SferH-1
and SferH-2 gene, the later forms the stable protein; hence,
SferH-2 gene was used to transform rice to enrich iron content.
To increase Fe content in rice seeds, the ferritin gene from Pha-
seolus vulgaris was expressed in rice endosperm under the
control of the glutelin promoter, resulting in a more than
twofold increase in the Fe content. The expression of recombi-
nant human lactoferrin (rHLF) in rice endosperm produced not
only 5 g rHLF per kilogram dehusked rice grains, but simulta-
neously increased Fe content about twofold. Similarly, expres-
sion of a heat-resistant Aspergillus fumigatus phytase resulted in
increased enzyme activity in the seed, but only a small residual
activity was detected after cooking. Phytase degrades phytate,
which is a Fe- and Zn-binding antinutritional compound.
In strategy II plants, there is a strong correlation between
the amount of phytosiderophores released and tolerance to
Fe deficiency. Graminaceous plants secrete natural Fe chelator
(mugineic acid phytoiderophores, MAs) from their roots to
solubilize Fe in the soil. These MAs are synthesized from L-
methionine through nicotianamine. NAS and NAAT are the
two critical enzymes in the biosynthesis of MAs, and in Fe-
tolerant species, activity of these two enzymes is increased
under Fe deficiency. Among strategy II plants, rice is particu-
larly sensitive to Fe deficiency when compared with barley.
Recently, barley genomic DNA fragments containing two
NAAT genes (NAAT-A and NAAT-B) that encode enzymes
involved in the biosynthesis of phytosiderophores were
successfully introduced in rice, using transformation tech-
niques. The resultant transgenic rice showed greater tolerance
to Fe deficiency and produced greater grain yields. These
results suggest there are possibilities for genetically engi-
neering Fe-efficient plants.
See also: Photosynthesis: C3 Plants. Plant Cells: Leaf
Development. Plant Nutrition: Deficiency Diseases, Principles;
Mineral Uptake; Nitrogen Fixation. Plants and the Environment:
The Rhizopshere and Its Microorganisms.
Further Reading
Abadia, J., Vazquez, S., Rellan-Alvarez, R., El-Jendoubi, H., Abadia, A., Alvarez-
Fernandez, A., Lopez-Milan, A.F., 2011. Towards a knowledge-based correction
of iron chlorosis. Plant Physiol. Biochem. 49, 471–482.
Barton, L.L., Abadia, J., 2006. Iron Nutrition in Plants and Rhizospheric Microorgan-
isms. Springer, Netherlands.

Briat, J.-F., Dubos, C., Gaymard, F., 2014. Iron nutrition, biomass production, and
plant product quality. Trends Plant Sci. 20, 33–40.
Curie, C., Cassin, G., Couch, D., Divol, F., Higuchi, K., 2009. Metal movement within
the plant: contribution of nicotianamine and yellow stripe 1-like transporters. Ann.
Bot. 103, 1–11.
Kim, S.A., Guerinot, M.L., 2007. Mining iron: iron uptake and transport in plants. FEBS
Lett. 581, 2273–2280.
Kobayashi, T., Nishizawa, N.K., 2012. Iron uptake, translocation, and regulation in
higher plants. Annu. Rev. Plant Biol. 63, 131–152.
Krohling, C.A., Eutropio, F.J., Bertolazi, A.A., Dobbss, L.B., Campostrini, E., Dias, T.,
Romas, A.C., 2016. Ecophysiology of iron homeostatis in plants. Soil Sci. Plant
Nutr. 62, 39–47.
Plant Nutrition j Iron Chlorosis 255
Lopez-Millan, A.-F., Grusak, M.A., Abadia, A., Abadia, J., 2013. Iron deficiency in
plants: an insight from proteomic approaches. Front. Plant Sci. 4. Article 254.
Mori, S., 1999. Iron acquisition by plants. Curr. Opin. Plant Biol. 2, 250–253.
Robinson, N.J., Procter, C.M., Connolly, E.L., Guerinot, M.L., 1999. A ferric-chelate
reductase for iron uptake from soils. Nature 397, 694–697.
Santi, S., Schmidt, W., 2009. Dissecting iron deficiency-induced proton exclusion in
Arabidopsis roots. New Phytol. 183, 1072–1084.
White, P.J., Broadley, M.R., 2009. Biofortification of crops with seven mineral
elements often lacking in human diets – iron, zinc, copper, calcium, magnesium,
selenium and iodine. New Phytol. 182, 49–84.