Agricultural

Communication

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

and

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

ABSTRACT

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.

716

ARTICLE INFORMATION:

*Corresponding Author:

Received 12

Oct, 2017

Accepted after revision 18

Dec, 2017

BBRC Print ISSN: 0974-6455

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DOI: 10.21786/bbrc/10.4/16

INTRODUCTION

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,

2015).

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,

2003).

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

BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS EFFECT OF DEFICIT AND ADEQUATE IRRIGATION AND NITROGEN FERTILIZER LEVELS 717

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.

MATERIAL AND METHODS

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.

RESULTS AND DISCUSSION

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

718 EFFECT OF DEFICIT AND ADEQUATE IRRIGATION AND NITROGEN FERTILIZER LEVELS BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS EFFECT OF DEFICIT AND ADEQUATE IRRIGATION AND NITROGEN FERTILIZER LEVELS 719

Hossein Heydari Sharif Abad et al

Table 1. AOVA table of data (2009 and 2010).

Mean squares (MS)

dfSource of variation

Fv/FmRWCLAICH C

0.0023.2030.59410.93Replication

0.00254.0282.584398.51Year

0.0012.4254.233.3583Year* replication

0.006**1312.09**4.871**293.136**2Water levels

0.0019.9150.4370.9962Year* water

0.00124.5560.076.15612Error

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

0.0017.0650.545.10836Error

5.784.719.166.9C.V.

Table 2. The effect of Water levels on Chlorophyll

content (Ch C), LAI, Relative water content (RWC), and

Fv/Fm.

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

Fv/Fm.

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

720 EFFECT OF DEFICIT AND ADEQUATE IRRIGATION AND NITROGEN FERTILIZER LEVELS BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

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).

CONCLUSION

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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