Biosci. Biotech. Res. Comm. 10(4): 716-721 (2017)
Effect of de cit and adequate irrigation and nitrogen
fertilizer levels on physiological traits of maize in
Kermansha province - Iran
Hossein Heydari Sharif Abad
, Mohamad Javad Mirhadi
, Ghorban Normohamadi
Afshin Charabeh
Department of Agronomy, Science and Research Branch, Islamic Azad University, Tehran, Iran
Ph.D. Student of Science and Research Branch, Islamic Azad University and Senior Expert of Agricultural
Bank, Tehran, Iran
In order to evaluation effects of drought stress and nitrogen levels on physiological traits of maize hybrid (Ksc703),
a  eld study was conducted at Kermanshah province, Western Iran, during 2009 and 2010. Effects of irrigation
(optimum, 85% and 60% of water requirement) and nitrogen (recommended, plus 25% and minus 25%) levels were
studied by using a split plot model and 4 replications. The LAI, chlorophyll content and Chlorophyll  uorescence
(Fv/Fm) were measured with a Sun Scan, chlorophyll meter (SPAD) and  uorimeter respectively. Chlorophyll content
and Fv/Fm was found to decrease with diminishing of available water, hence increase of N resulted in increase of
chlorophyll content and Fv/Fm in both normal and stress conditions. Drought stress reduced LAI in all N levels. The
increase of N increased LAI in normal condition. In contrast LAI increased by increase of N usage, but in high level
of N (more than recommended rate) LAI reduced. Drought stress reduced RWC, high level of N reduced RWC also.
*Corresponding Author:
Received 12
Oct, 2017
Accepted after revision 18
Dec, 2017
BBRC Print ISSN: 0974-6455
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Online Contents Available at:
DOI: 10.21786/bbrc/10.4/16
Maize (Zea mays L.) is an important crop that used as
food, feed and industrial products. Maize is the third
most important cereal after wheat and rice all over the
world and the world’s largest grain crop in term of total
production on a MT basis. Maize planting area is about
184 million hectares in 125 countries and is the most
important crop in 75 countries (FAOSTAT, 2015). Maize
is one of the most important crops in the western part
of Iran there is a shortfall in the production of animal
feeds. Kermanshah is located in western Iran and Maize
is the most important crop after wheat, grown on an
area of 45,000 ha with the production of 382,500 tones
Hossein Heydari Sharif Abad et al.
with 8500 kg ha-1 average grain yield; the third and
rst place of Iran for area harvested and mean yield,
respectively. The mean annual precipitation in Iran is
240 ml and it is seen as a dry or semidry country. Maize
is an irrigated crop in Iran and recent drought periods
in Iran imposed pressure on groundwater resources. The
groundwater is the primary source of irrigation of maize
production in province and in recent years, the ground-
water levels has gradually decreased in this region
mainly because of increasing annual irrigation and the
dry climate (Agricultural Department of Kermanshah,
The dearth of water is one of the major factors chal-
lenging maize production. Among agro-meteorological
hazards, drought has the greatest effect on yield stability
(Vinocur and Altman, 2005). It can seriously affect the
grain quality and grain output reducing average yields
by 50% or even more (Wang et al., 2003). Water short-
age due to decreasing annual precipitation and the dry
climate, as well as low fertility and low percentage of
organic matter in soil is major problems of maize pro-
duction in Iran. For this reason, overuse of chemical fer-
tilizers in Iran is rising, leading to environmental pollu-
tion and soil degradation. Yield losses include more than
two- thirds of the total damage of abiotic stresses due to
drought, salinity and other factors. Maize is highly sen-
sitive to drought stress, speci cally in  owering stage
(Tollenaar and Lee, 2011).
Drought is a major problem for the production of the
world’s  ve principal cereals: maize, wheat, rice, pearl
millet, and sorghum. Water stress reduced yield in crop,
but interactive effects of water and nitrogen de cits on
physiological traits and on physiological changes asso-
ciated with leaf aging have received little attention (Val-
liyodan and Nguyen 2006). Soil water de cit reduces
yield of maize (Zea mays L.) and other grain crops by
three main mechanisms. First, whole canopy absorption
of incident Photosynthesis Active Radiation (PAR) may
be reduced, either by drought induced limitation of leaf
area expansion, by temporary leaf wilting or rolling dur-
ing periods of severe stress, or by early leaf senescence.
Second, drought stress reduces the ef ciency with which
absorbed PAR is used by the crop to produce new dry
matter. Third, drought stress may limit grain yield of
maize by reducing the harvest index (Earl and Davis,
Mihailovic et al (1992) demonstrated one of the fac-
tors in uencing physiological responses of plants to
water stress is mineral nutrition. A signi cant role of
nitrogen in regulating plant responses to water stress
was established in a number of plant species. According
to estimates of CIMMYT (International Maize and Wheat
Improvement Centre) regarding abiotic stresses, the most
signi cant causes of yield loss on farmers  elds are
low fertility (predominantly N de ciency) followed by
drought and, less important, by plant competition related
to low planting densities, weeds and intercrops (Ribaut
and Poland, 1999). Some of the effects of drought stress
on physiological traits of plants are suitable. Photosyn-
thetic rate, leaf surface area and photosynthetic capacity
enhanced with increase in nitrogen levels (Gungula and
et al, 2005). Leaf area and LAI increase with increase in
nitrogen levels (Oscar and Tollennar, 2006).
The chlorophyll meter (or SPAD meter) is a simple,
portable diagnostic tool that measures the greenness or
the relative chlorophyll concentration of leaves. Com-
pared with the traditional destructive methods, this
equipment might provide a substantial saving in time,
space and resources. The Minolta Soil Plant Analy-
sis Development (SPAD-502) chlorophyll meter is one
tool that enables researchers to determine chlorophyll
content by measuring leaf greenness (Peterson et al.,
1993). The SPAD uses a silicon photodiode to derive the
ratio of transmittance through the leaf tissue at 650 nm
compared with transmittance at 940 nm, and a value
is given based on that ratio. SPAD measures relative
chlorophyll content in plant leaves. Because chloro-
phyll content is closely related to N supply (Pandey et
al., 2000), SPAD is also used to diagnose maize N sta-
tus and predict maize grain yield potential (Vetsch and
Randall, 2004). Janos (2010) reported a close correlation
between N fertilization and SPAD readings. Increasing N
application increased N content and chlorophyll content
in maize (Rambo et al., 2010). Factors affecting SPAD
values include radiation differences between seasons,
variety and species differences, plant and soil nutrient
status (including N and other nutrients), and biotic and
abiotic stresses (Peterson et al., 1993).
Atteya (2003) showed exposure of plants to drought
lead to noticeable decrease in leaf water potential and
relative water content (RWC) and Water stress changed
the relation between leaf water potential and relative
water content of maize so that stressed plants had lower
water potentials than control at the same leaf RWC. The
RWC measurement characterizes the internal water sta-
tus of plant tissues and is also a convenient method for
following changes in tissue water content without errors
caused by continually changing tissue dry weight (Erick-
son et al., 1991). On the other hand using the chlorophyll
uorescence technique, is useful possible to estimate the
parameters of actual photosynthetic ef ciency of leaf,
under various conditions at various times, and also the
potential maximum of the quantum ef ciency (Fv/Fm).
Fv/Fm is the measurement of quantum yield potential
of photosynthesis, or maximal photochemical ef ciency
of PSII. The Fv/Fm ratio has been shown to be a reliable
Hossein Heydari Sharif Abad et al.
indicator of stress (Duraes and et al, 2001). Photosyn-
thesis, as a signi cant physiological process to yield is
sensitive to water stress. The photosynthetic rate keeps
decreasing while the intension of stress increases, which
is the main reason for the reduction of yield by drought,
Moreover, it is possible to determine if there is damage
to light reaction systems in photosynthetic machinery
during drought (Liu et al., 2012). Measuring chlorophyll
uorescence has become a very useful technique in
obtaining rapid qualitative and quantitative information
on photosynthesis (Rohácek, 1999), and it can provide
information on the relationship between structure and
function of photosystem II (PSII) reaction center (Rosen-
qvist and Van Kooten, 2003). Chlorophyll  uorescence
provides useful information about leaf photosynthetic
performance of many plants under drought stress (Baker
and Rosenqvist, 2004).
Schlemmer et al (2005) reported a very strong rela-
tionship between the Minolta SPAD-502 chlorophyll
meter readings and direct measurements of chlorophyll
content in maize and soybean leaves. Since chlorophyll
content is usually strongly related to N concentration,
these meters can be used as indicators of need for agri-
cultural N application. So, present research conducted in
Kermansh, a drought stress prone province, to study of
physiological aspects of water de cit and nitrogen levels
on growth and development of maize hybrids.
In order to evaluation effects of drought stress and nitro-
gen levels on physiological traits of maize hybrids, a
eld study conducted in Kermanshah province, western
Iran, at 2016 and 2017 at the agricultural research farm,
Agricultural and Natural Resources Research Centre in
Kermanshah, Iran. This farm is located at 34.08 N, 46.26
E, 1345 m altitude, silty clay soil, pH=7.5-8, 450 mm
precipitation Mediterranean climate.. The KSC703 (late
maturating group) maize hybrid is dominate commercial
cultivar of Kermansha province. Experiment plots were
seeded with 75 cm row to row distance and plant density
was 75000 plant/ha (conventional plant density). Seeds
were sown 7 cm deep. Maize was planted in May and
by experimental planter. Irrigation (optimum, 85% and
60% of water requirement) and nitrogen levels (recom-
mended, plus 25% and minus 25%) arranged as main
and sub plots respectively using a complete randomized
block and 4 replication were used. LAI was measured
with a Sun Scan canopy analysis system (Delta-T
Devices, Cambridge, UK) and in stages V6, V10 and R1.
Chlorophyll meter (SPAD-502, Minolta) readings were
taken in all plots. SPAD reading were taken on the mid-
point of the youngest fully expanded leaf.
Chlorophyll  uorescence (potential maximum of the
quantum ef ciency (Fv/Fm)) in leaves of non-stressed
and water stressed plants was measured in R1 stage
with chlorophyll  uorimeter (Pocket PEA). The RWC
was measured using  ag leaves after imposing drought
conditions. Leaves were sealed within plastic bags and
quickly transferred to the lab. Fresh weight (FW) was
determined within 2 h after excision. Turgid weight
(TW) was obtained after soaking leaves in distilled water
in test tubes for 16 to 18 h at room temperature. After
soaking, leaves were quickly and carefully blotted dry
with tissue paper in preparation for determining turgid
weight. Dry weight (DW) was obtained after oven drying
the leaf samples for 72 h at 70ºC. RWC was calculated
from the formula:
RWC (%) = [fresh weight- dry weight/ turgid weight
– dry weight] × 100
Where FW, TW and DW are fresh weight (g), turgid
weight (g) and dry weight (g) respectively.
Analysis of data showed that chlorophyll content at tas-
seling decreased by diminish of available water, hence
increase of N, resulted in increase of chlorophyll con-
tent in both normal and stress condition. A direct close
relationship of nitrogen levels with SPAD (Chlorophyll
Meter Readings) was reported in maize (Schlemmer and
et al 2005). Ciganda et al (2008) had similar results and
reported that chlorophyll content is among the most
important crop biophysical characteristics. Chlorophyll
can be related to photosynthetic capacity, thus, produc-
tivity, developmental stage, and canopy stresses, also
Munne-Bosch and Alegre (2000) reported the chloro-
phyll content was decreased with decreasing the irriga-
tion water and this decrease was correlated with relative
water content in leaves. Chlorophyll loss is a negative
consequence of water stress.
Drought stress reduced LAI in all N levels. The increase
of N increased LAI in normal condition. In contrast LAI
increased by increase of N use, but in high level of N
(more than recommended rate) LAI reduced. Similarly
in accordance with our results, LAI was positively cor-
related with nitrogen application in normal condition
(Oscar and Tollennar, 2006). For most plant species, the
shortage of nitrogen or water causes a reduction in leaf
area development, changes in leaf tissue composition,
leaf cell structure and plant water content (Casa, 2003)
and also in maize, drought reduces leaf area, leaf chlo-
rophyll contents, photosynthesis and ultimately lowers
the grain yield (Athar and Ashraf, 2005). Stone et al.
(2001) reported that water de cit reduces crop growth
Hossein Heydari Sharif Abad et al
Table 1. AOVA table of data (2009 and 2010).
Mean squares (MS)
dfSource of variation
0.0012.4254.233.3583Year* replication
0.006**1312.09**4.871**293.136**2Water levels
0.0019.9150.4370.9962Year* water
0.00141.383**0.56**325.969**2Nitrogen levels
0.0010 ns0.156 ns0.32 ns8.211 ns2Year* Nitrogen
0.001 ns19.202*1.71**2.091 ns4Water* Nitrogen
0.0011.9770.0760.976 ns4Water * Nitrogen * Year
Table 2. The effect of Water levels on Chlorophyll
content (Ch C), LAI, Relative water content (RWC), and
Water levels CH C LAI RWC Fv/Fm
Optimum (100%) 41.44 3.03 79.15 0.81
80% requirement 38.96 2.60 71.66 0.789
60% requirement 34.54 2.13 64.36 0.781
Table 3. The effect of Nitrogen levels on Chlorophyll
content (Ch C), LAI, Relative water content (RWC), and
Nitrogen levels CH C LAI RWC Fv/Fm
Recommended - 25% 34.45 2.49 73.02 0.78
Recommended 38.45 2.51 71.76 0.79
Recommended + 25% 41.8 2.06 70.39 0.80
Table 4. The effect of Nitrogen and water levels on Chlorophyll content (Ch C),
LAI, Relative water content (RWC), and Fv/Fm.
Water levels Nitrogen levels CH C LAI RWC Fv/Fm
Optimum (100%)
Recommended - 25% 37.6 2.69 78.8 0.80
Recommended 41.83 2.9 79 0.81
Recommended + 25% 44.9 3.7 79.5 0.82
80% requirement
Recommended - 25% 34.9 2.4 74 0.78
Recommended 38.9 2.7 72.1 0.79
Recommended + 25% 43.03 2.5 68.1 0.79
60% requirement
Recommended - 25% 30.84 2.3 66 0.76
Recommended 35.33 2.1 64.1 0.76
Recommended + 25% 37.46 1.9 62.8 0.77
and morphological characteristics of maize plant. In
maize, reproductive growth after the silking and  ower-
ing stages is the critical period for yield, and chlorophyll
content and intact chloroplast structure are key fac-
tors for accumulation of dry matter and high yields (Yu
et al., 2010).
The RWC measurement characterizes the internal
water status of plant tissues and is also a convenient
method for following changes in tissue water content
without errors caused by continually changing tissue
dry weight. Drought stress reduced RWC. The RWC was
80 to 64 for normal and drought stress condition respec-
tively. High level of N reduced RWC also. Jabasingh and
Saravana Babu (2014) had similar results and reported
that the relative water content in leaves of different
maize cultivars decreased signi cantly and with drought
stress, the membrane permeability of the leaf cell mark-
edly increased. Also Higher RWC indicates better growth
Hossein Heydari Sharif Abad et al
and development, which in turn depends on leaf area
(Sivakumar, 2014).
The results showed that with reduction of water
availability, the quantum ef ciency (Fv/Fm) decreased.
On the other hand with increase of nitrogen level, Fv/
Fm increased but not statistically signi cant. Duraes
et al (2001) reported that the Fv/Fm will reduce by drought
stress in maize hybrids. Photochemical chlorophyll  uores-
cence quenching, photosystem II quantum yield and elec-
tron transport rate and more heat dissipation as compared
to controls (Dias and Bruggemann, 2010). Light energy
absorbed by chlorophyll molecules in a leaf can undergo
one of three fates: it can be used to drive photosynthe-
sis (photochemistry), excessive energy can be dissipated as
heat or it can be re-emitted as light-chlorophyll  uores-
cence. These three processes occur in competition that is
any increase in the ef ciency of one will result in a decrease
in the yield of the other two (Maxwell and Johnson, 2000).
Results showed that under drought stress RWC, chloro-
phyll content, LAI and the quantum ef ciency (Fv/Fm)
decreased. Therefore, reducing of RWC, LAI and Chlo-
rophyll content or quantum ef ciency (Fv/Fm) could be
indicative of water stress. On the other hand increase
of nitrogen, resulted in increase of chlorophyll content
and Fv/Fm in both normal and stress conditions and
with increasing amounts of nitrogen up to 2
level, LAI
increased, but RWC decreased.
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