Journal of Maize Research and Development (2015) 1(1):134-145

ISSN: 2467-9283 (Print)/ 2467-9291 (Online)

Analysis of chlorophyll content and its correlation with

yield attributing traits on early varieties of maize (Zea
mays L.)
Bikal Ghimire*1, Deepika Timsina 1 and Jyotsna Nepal1

ARTICLE INFO

Article history:
Received:
5th April, 2015
Revised:
7th September, 2015
Accepted:
6th November, 2015
Keywords:
Maize, correlation
coefficient,
chlorophyll content

ABSTRACT
Chlorophyll has direct roles on photosynthesis and hence closely relates to capacity
for photosynthesis, development and yield of crops. With object to explore the roles
of chlorophyll content and its relation with other yield attributing traits a field
research was conducted using fourteen early genotypes of maize in RCBD design
with three replications. Observations were made for Soil Plant Analysis
Development (SPAD) reading, ear weight, number of kernel row/ear, number of
kernel/row, five hundred kernel weight and grain yield/hectare and these traits were
analyzed using Analysis of Variance (ANOVA) and correlation coefficient analysis.
SPAD reading showed a non-significant variation among the genotypes while it
revealed significant correlation with no. of kernel/row, grain yield/hectare and
highly significant correlation with no. of kernel row/ear and ear weight which are
the most yield determinative traits. For the trait grain yield/ha followed by number
of kernel row/ear genotype ARUN-1EV has been found comparatively superior to
ARUN-2 (standard check). Grain Yield/hectare was highly heritable (>0.6) while
no. of kernel / row, SPAD reading, ear weight, number of kernel row/ear were
moderately heritable (0.3-0.6). Correlation analysis and ANOVA revealed ARUN1EV, comparatively superior to ARUN-2(standard check), had higher SPAD
reading than mean SPAD reading with significant correlation with no. of
kernel/row, no. of kernel row/ear, ear weight and grain yield/ha which are all yield
determinative traits . This showed positive and significant effect of chlorophyll
content in grain yield of the maize.

INTRODUCTION
Maize (Zea mays L.) is one of the most important cereal crops of the world. It has remarkable
production potential in Nepal and is the second staple food in Nepal after paddy. After paddy,
maize is important cereal crop of Nepal after rice in terms of area, production and
productivity (MoAD, 2013/14). At present, the Maize sown area in Nepal is 9,28,761 ha
and total production is 22,83,222 metric tons with productivity of 2458 Kg/ha (MoAD,
2014). In Chitwan, maize covers total area of 9750 ha with the total production of 29,250
metric tons and productivity of 3,000 Kg/ ha (MoAD, 2014). Chitwan is a terai area and out
of total maize cultivated area in Terai region, 95.95 % area is under
Corresponding author Info:
1

Tribhuwan University, Institute of Agriculture and Animal Science (IAAS),

Rampur Campus, Rampur, Chitwan, Nepal
* Email: [email protected]
Contact number: +9779845180818

135
improved and 4.05 % area under local maize. Maize exhibit sequential order of yield
components development namely number of ear/plant, number of kernel/row, number of
kernel row/ ear and hundred kernel weights (Viola et al., 2004). There has been various study
by different scholars about the yield attributing traits in maize. Among them chlorophyll content
is one of the important attributing trait as it relates to photosynthesis capacity and stages of crop
development. It is an important matter to study chlorophyll content and its significant roles in
yield and production of maize in developing country like Nepal where there has not been much
study in molecular level in case of maize and maize breeding. Chlorophyll accounted for more
than 98% of gross primary production variation in maize (Gitelson et al., 2008). The chlorophyll
content of leaf has been suggested as the community property and has proportional relation to
predict productivity. Not much information about the combining ability and genetic parameters
about chlorophyll content of three-ear-leaves has been available yet. However, to evaluate the
combining ability, reciprocal effect and genetic parameters for chlorophyll content of three-earleaves and how is it correlated with maize yield and other yield attributing traits was the
objective of this research. By measuring the chlorophyll content, we can indirectly assess the
potential taking up infrared radiation and the ability of a leaf to stay green (Araus et al.,
2008). The ratio of absorbance of radiation at 650 nm (chlorophyll absorbance peak) and at
940 nm (non-chlorophyll absorbance) can be calculated by Soil Plant Analysis Development
(SPAD) meter SPAD-502. SPAD reading shows a positive relation with yield in maize as a
result of transport of energy from photosynthesis due to increased production or shows negative
relation with yield if energy is remobilized from chlorophyll. Any crop improvement is
influenced by both genotypic variation and phenotypic variation. Degree of variability is an
essential tool for effective selection. The phenotypic variation is affected by genotypic variation,
environmental variation and interactions of both kinds (Lee & Lamkey, 2006). The total variance
of a given character is its phenotypic variance (2p) and environmental variance (2e) attributed
to environmental conditions. Determination of genotypic and phenotypic correlation is very
fundamental step in the formulation and implementation of various breeding programs and
activities. Correlation measures the degree of association, genetic or non-genetic, between two or
more characters and is measured by a correlation coefficient (Hallauer & Miranda, 1988). In
plant breeding, generally two types of correlations are discussed viz. genotypic correlation and
phenotypic correlation. Phenotypic correlation (rp) encompasses both genetic and environmental
effects while the association of breeding values (i.e., additive genetic variance) of the two
characters is genotypic correlation. Both of the correlations measure the extent how closely
linked genes cause co-variation in two different characters (Hallauer & Miranda, 1988).The
study was conducted to determine how chlorophyll content of three ear leaf plays role in
maize yield and how it is correlated with other yield attributing traits. The research was
conducted by taking fourteen genotypes including one standard check variety with following
objectives:
To study the chlorophyll content and its roles on maize productivity
To study the correlation between chlorophyll content and other yield attributing
traits of maize.

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MATERIALS AND METHODS
Research site
Field experiment was conducted at research field of National Maize Research Program
(NMRP) from 2 2 n d October 2014 to 12th April, 2015. The location is at 2737 N latitude and
8425E longitude with an elevation of 228 meters above sea level (NMRP, Rampur).
Climatic condition and season
The research site was climatically humid and sub-tropical. The average annual rainfall was
2000 mm (NMRP, Rampur). The meteorological data (Figure 1) were obtained from
meteorological station, National Maize Research Program, (NMRP), Rampur.
Experimental details/materials/selection of genotype
Randomized Complete Block Design (RCBD) was laid with three replications and fourteen
maize genotypes as treatments. These genotypes were allocated randomly to the fourteen plots
of each replication. The plot size was 5.0 m 3m with inter row spacing of 75cm and and intra
row spacing of 25 cm, respectively. Each genotype was sown in each plot with four rows each
of 5 m length. All genotypes were obtained from NMRP, Rampur. Following are the
genotypes used in the research:
Entry no.

Genotype

Entry no.

Genotype

1.

ACROSS- 2401

8.

ARUN-2(standard check)

2.
3.
4.
5.
6.
7.

RAJAHAR LOCAL
S97TEYGHAYB(3)
POP-445/POP-446
ARUN-1EV
R.C/POOL-17
SO3TETEY/LN

9.
10.
11.
12.
13.
14.

FARMERS VARIETY
ZM-621/POOL-15
EEYC1
KY/Pool-17
Pool-27
Pool-15

Crop management
Land preparation was performed by ploughing two times followed by land leveling. Farm yard
manure (FYM) was applied at the rate of 10 tons/ha and chemical fertilizers as per general
recommendation for maize were applied at the rate of 120:60:40 NPK kg/ha of Nitrogen,
Phosphorus and potassium respectively. Full dose of Phosphorus, Potassium and half dose of
Nitrogen were applied at the time of sowing as basal application. Sowing was done on 22nd
October, 2014 by manually operated sowing machine. Other half dose of Nitrogen was applied in
splits of two doses doses during first and second weeding. Two hand weedings were carried out at
20-25 DAS. Thinning was done during first 15- 20 DAS. Earthing up was done as second
weeding. The maize borer was controlled by placing granules of Phorate in top folding leaf.
Phorate is an organophosphate used as an insecticide and acaricide.
Data collection
All the observation was recorded from five randomly selected plants of each plot and average

137
values were taken for analysis. Observations were taken for major traits viz. SPAD measures,
number of kernels per row, number of kernel rows per ear, ear weight (gm), 500 kernel weight
(gm) and grain yield per hectare with moisture adjustment at 14%.
Chlorophyll Nitrogen implying SPAD measures (SPAD reading)
Chlorophyll content o f l e a f was measured by using Soil Plant Analysis Development (SPAD)
meter (SPAD 502, Minolta, Japan). Five plants were randomly selected from each plot to
measure chlorophyll content.
Number of kernels/row
Number of kernels/ row was counted and recorded for five randomly selected ear and
average value was taken.
Number of kernel row/ear
Number of kernel rows/ ear was counted and recorded for five randomly selected ear and
average value was taken.
Ear Weight (gm)
Ear weight was measured after harvesting the cob and its weight was measured for individual
cob.
Five-Hundred kernel weights (g)
Five Hundred kernel weights of samples was recorded along with their moisture content by
using moisture meter and then it was converted to fourteen percent (14%) moisture
content by using formula:
100 Moisture content %
500-Kernel weight= ----------------------------------------- (500-kernel weight)
100-Required moisture% (14%)
Grain yield
To calculate grain yield, yield/plot was converted into grain yield/ ha. Grain Yield/ plot was
converted to 14% moisture by using formula as above and then it was converted to grain
yield/ha at 14% moisture by using given formula:
Yield / plot at 14% moisture (kg)
10000 m2

GY=
Plot size in m2

Phenotypic and genotypic variance

Formula given by (Lush, 1940) and (Chaudhary & Prasad, 1968) was used to calculate
phenotypic and genotypic variance.
Genotypic variance = 2g = (TMSS EMSS) / R
Error variance = 2e
Phenotypic variance = 2p = 2g + 2e
Where,

138
TMSS is treatment mean sum of square
EMSS is error mean sum of square
R is number of replication
Genotypic and phenotypic coefficient of variation (GCV and PCV)
They are expressed as percentage. According to (Burton & Devane, 2008):
Genotypic coefficient of variation (GCV) = (g/X) 100
Phenotypic coefficient of variation (PCV) = (p/ X) 100
Where,
g = Genotypic standard deviation
p = Phenotypic standard deviation
X = General mean of the trait
As indicated by (Sivasubramanjan & Menon, 1973), GCV and PCV are categorized as
follows:
0 10 %
: Low
10 20 % : Moderate
>20 %
: High
Broad sense heritability ( h b s 2 )
Hanson et al ., (1956) estimated broad sense heritability as the ratio of genotypic
variance (Vg) to the phenotypic variance (Vp) and expressed in percentage.
Broad sense heritability (h b s 2 ) = (V g /V p )
Robinson et al., (1949) categorized broad sense heritability as follows:
0 0.30
: Low
0.30 0.60
: Moderate
> 0.60
: High
Genetic advance (GA)
It was calculated by using the following formula given by (Robinson et al., 1949).
GA = i. p.hbs2
Where,
i = Efficacy of selection (2.06 at 5% selection intensity)
p = Phenotypic standard deviation
hbs2 = Broad Sense Heritability
Genetic advance as percent of means (GAM)
GA as per cent of mean (GAM) = (GA/X) 100
GA = Genetic advance
X =General mean of the trait
Johnson et al., (1955) categorized GAM as follows:
0 - 10 %
: Low
10 -20 %
: Moderate
> 20 %
: High
Statistical analysis

139
Analysis of variance was carried out by using statistical software GENSTAT, MINITAB version
20 as well as correlation analysis was done using SPSS version 21.

RESULTS AND DISCUSSION

Mean performance and analysis of variance
Mean values and significant levels of yield and yield attributing traits of fourteen maize
germplasm are presented in Table 1. Significant results were observed for the for the traits
grain yield/ha, ear weight, number of kernel/row, number of row kernel / ear, while non
significant result was observed for SPAD reading and five hundred kernel weight.
Chlorophyll nitrogen implying SPAD measures (SPAD reading)
A non-significant difference (P0.05) in SPAD was found for the genotypes (Table 1).
Maximum SPAD (51.74) had been showed by ARUN-2 (standard check) and minimum SPAD
(44.59) had been showed by Pool-27.
Number of kernel rows/ear (NKRE)
The result showed significant difference (P0.05) in NKRE for the genotypes (Table 1).
Maximum NKRPE (13.07) has been shown by ZM-621/POOL-15 and minimum (9.60) has been
shown by KY/Pool-17.
Number of kernel/row (NKPR)
The result showed significant difference (P0.05) in NKPR for the genotypes (Table 1).
Maximum NKPR (23.67) has been showed by FARMERS VARIETY and minimum (15.07) has
been showed by KY/Pool-17. Variety S97TEYGHAYB(3), FARMERS VARIETY, ZM621/POOL-15 , RAJAHAR
LOCAL, POP-445/POP-446, ARUN-1EV, R.C/POOL-17,
SO3TETEY/LN , EEYC1, Pool-27, Pool-15 were statistically similar to ARUN-2 (standard
check) (Table 1).
Five Hundred kernel weights (FHKW)
The result showed that there was non-significant difference (P0.05) in FHKW for the genotypes
(Table 1). Maximum FHKW (150.7) has been showed by ACROSS-2401 and minimum (112.0)
has been showed by RAJAHAR LOCAL.
Grain yield Kg/Hectare (GYPH)
The result showed that there was significant difference (P0.05) in the grain yield kg/ha for the
genotypes (Table 1). Among the tested genotypes ARUN-1EV has been found high yielder with
grain yield of 2376 kg/ha and genotype KY/pool-17 with 164 kg/ha was low yielder genotype
(Figure 2). The genotypes ARUN-1EV, S97TEYGHAYB (3), Farmers variety , SO3TETEY/LN,
Pool-15, POP-445/POP-446 were found statistically at par with ARUN-2 (standard check) for
grain yield kg/ha. The grain yield/ha in relation to genotypes has been shown in figure 2.
Broad sense heritability (hbs2)
The considerable differences in heritability value for different characters were observed (Table
2). Among quantitative characters, high heritability (>0.60) has been observed for grain yield/ha

140
(0.67), Moderate heritability (0.30 to 0.60) has been found for number of kernel/row (0.344),
SPAD reading (0.118), ear weight (0.45), number
of kernel row/ear (0.376) while low
heritability (<0.30) has been observed in five hundred kernel weight. (Viola et al., 2004) reported
high heritability for grain yield/ha which is very consistent with our finding. Grain yield/ha and
ear height had better genotypic variability; better broad sense heritability along with better GA is
considered the good estimates for effective selection of a trait. This depicted that visual selection
based on these traits among the genotypes would be used for improvement of grain yield.
Genetic advance as percentage of mean (GAM)
GAM at five percent selection intensity exhibited greater differences for quantitative characters
as represented in Table 2. High GAM having value more than 20% were estimated for ear weight
and grain yield/ha. GAM between 10 and 20 % has been observed for number of kernel / row.
Lower values were estimated for five hundred kernel weights, number of kernel row/ear, SPAD
reading and ear length. GAM showed that ear weight, plant height and grain yield/ha were under
control of additive genes. Alvi et al., (2003) reported similar findings.
Phenotypic Coefficient of Variation and genotypic coefficient of variation (PCV and GCV)
The considerable difference in PCV and GCV values for different traits has been observed
(Table 2). Among the studied quantitative traits, PCV and GCV values were estimated higher for
grain yield/ha, ear weight had high PCV and moderate GCV, whereas five hundred kernel
weights, number of kernel/row had moderate PCV values And number of kernel row/ear, SPAD
reading had lower PCV and GCV value. The difference between GCV and PCV ranged from
3.747 to 10.913 and was lower for all traits studied. This showed the higher genetic effects in
these parameters for variation. Higher broad sense heritability of all traits revealed that larger
portion of variations is heritable to offspring.
Correlation coefficient
Analysis of correlation coefficient of yield related traits revealed some fundamental basis.
Regression equation showed the relationship between the grain yield/ha with variables i.e. SPAD
reading, number of kernel row/ear, number of kernel/row, ear weight and five hundred kernel
weights. All traits showed positive correlation with grain yield/ha (Figure 3). This means that
grain yield per hectare increases with increase in value of SPAD reading, number of kernel
rows/ear, number of kernel/row, ear weight and five hundred kernel weight. Ibitome (2010) and
Rafique et al. (2004) reported similar findings. Chlorophyll content by SPAD reading was found
significantly correlated with number of kernel/row and number of kernel row/ear, ear weight
were highly significant with SPAD reading. Thus, these traits have highly significant correlation
with grain yield / hectare. The high direct effect of SPAD reading, number of kernel row/ear,
number of kernel/row and ear weight appeared to be the main factor for their strong association
with grain yield/plant. Hence direct selection for these traits would be effective. The traits grain
yield/ha, number of kernel/row have shown higher heritability and also expressed highly positive
and significant correlation coefficient. Thus, these traits are to be considered in selecting
genotypes for better crop improvement and breeding programs.
CONCLUSION
Chlorophyll content and its correlation with yield attributing traits among the different maize
genotypes was observed from this study which is very useful for their improvement of

141
agronomically important traits. Grain yield is the very complex parameter in Maize. Any minor
change in any component leads to the yield loss. Grain yield and related traits are very sensitive
to any crops. Association of different yield and yield attributing traits are very important to know
their direct and indirect effects on grain yield. In this study we emphasized to determine the
correlation coefficient of the traits with chlorophyll content in order to understand and identify
how chlorophyll play a vital role in selection and breeding for simultaneous improvement of
genetic materials. For the trait grain yield ha-1 followed by number of kernel row/ear, ARUN1EV has been found comparatively superior to ARUN-2(standard check). For yield attributing
traits like number of kernel row/ear, number of kernel/row and five hundred kernel weight
genotype SO3TETEY/LN, FARMERS VARIETY and ACROSS-2401 showed highest value
respectively. Grain Yeild Per hectare was highly heritable .Similarly, SPAD reading, no. of
kernel per row, number of kernel row per ear, ear weight were moderately heritable. Higher
GCV and high GAM indicate efficient indirect selection for higher grain yield/ha based on these
traits. Thus high GAM and GCV was observed in ear weight ,grain yield per hectare and ear
height. Correlation analysis revealed that the traits number of kernel row per ear , no. of kernel
per row and ear weight were the most yield determinative traits and hence, selection of these
traits for further breeding program might bring an improvement in grain yield. SPAD reading for
chlorophyll content showed positive significant correlation with grain yield. Correlation analysis
and ANNOVA revealed ARUN-1EV, comparatively superior to ARUN-2 (standard check), had
higher SPAD reading than mean SPAD reading with significant correlation with no. of
kernel/row, no. of kernel row/ear, ear weight and grain yield/ha which are all yield determinative
traits . This showed positive and significant effect of chlorophyll content in grain yield of the
maize. Besides the correlation inter se associations also provide huge support on these six traits
from all other yield related components. The variability shown by different genotypes for
different yield attributing quantitative traits can be used for the development of the high yielding
and better performing variety. From the research, selection of ARUN-1EV genotype was found
to be reliable for further research and breeding program in early varieties of maize. SPAD
reading exhibiting chlorophyll content was found significantly correlated with grain yield and
yield attributing traits.
Table 1.Means different parameters of fourteen maize genotypes in Chitwan, Nepal, 2014/15

Genotype
ACROSS- 2401
RAJAHAR LOCAL
S97TEYGHAYB(3)
POP-445/POP-446
ARUN-1EV
R.C/POOL-17
SO3TETEY/LN
ARUN-2(std check)
FARMERS VARIETY
ZM-621/POOL-15
EEYC1
KY/Pool-17
Pool-27

SPAD
47.81
47.17
45.37
48.01
48.53
45.27
50.00
51.74
49.59
48.96
46.96
46.01
44.59

Mean
NKRPE
11.60a
12.40a
12.67a
12.67a
12.13a
11.60a
12.53a
11.60a
11.33a
13.07a
12.40a
9.60b
11.33a

NKPR
17.53bc
19.80ab
23.13a
20.73ab
21.67ab
19.33abc
21.13ab
23.33a
23.67a
20.53ab
21.93ab
15.07c
20.93ab

EW(gm)
64.7bcd
61.4cd
81.1abc
76.1abc
86.0ab
61.1cd
87.2ab
93.9a
88.7ab
73.2abc
81.3abc
49.2d
64.7bcd

FHKW(gm)
150.7
112.0
120.7
122.0
134.0
116.7
138.7
140.7
132.7
117.3
126.0
119.0
123.3

GYPH(kg)
493ef
728def
2222a
1922abc
2376a
711def
2175ab
2232a
1982ab
820def
1073cdef
164f
1278bcde

142

Pool-15
Grand mean
SEM ()

12.67a
11.97
0.529

51.58
47.97
1.907

20.67ab
20.68
1.434

88.3ab
75.5
7.20

1598abcd
1412
281.5

130.0
127.4
9.97

LSD0.05
ns
1.537*
4.169*
20.92**
ns
818.4**
CV %
6.9
7.7
12.0
16.5
13.5
34.5
*(significant), **(highly significant) & ns(non significant) at p=0.05. Treatment means bearing same letter are not
significant different at p=0.05 by DMRT. SEM=Standard Error of Mean, LSD=Least Significant Difference &
CV=Coefficient of Variance, SPAD=Spad Reading, NKRPE=Number of Kernel Row / Ear, NKPR=Number of
Kernel / Row, EW=Ear Weight, FHKW=Five Hundred Kernel Weight, GYPP=Grain Yield / Plant, GYPH=Grain
Yield / Hectare.
Table 2: Estimation of genetic parameters ( GAM, PCV and GCV) for fourteen maize genotypes in Chitwan,
Nepal, 2014/15
Variable

GCV

PCV

SPAD
NKRPE
NKPR
EW
FHKW
GYPH

2.519
5.941
8.7
14.922
3.43
49.193

7.332
9.688
14.831
22.255
13.977
60.106

hbs2

GAM

0.118
0.376
0.344
0.45
0.06
0.67

1.783
7.506
10.513
20.612
1.734
82.939

Vg=genotypic variance, Ve=environmental variance, Vp=phenotypic variance, GCV=genotypic coefficient of

variation, PCV=phenotypic coefficient of variation, GAM=genetic advance as percentage of mean, h bs2=broad sense
heritability, SPAD=Spad reading, NKRPE=number of kernel row/ear, NKPR=number of kernel/row, EW= Ear
Weight, FHKW= Five hundred kernel weights, GYPH= grain yield/ha
Table3. Pearsons correlation coefficient between yield and yield attributing traits of fourteen genotypes of
maize in Chitwan, Nepal, 2014/15
NKRE

NKPR

EW

FKW

SPAD
NKRE
NKPR
EW
FKW

SPAD
1.00
0.45**
0.31*
0.51**
0.04ns

1.00
0.53**
0.58**
-0.11ns

1.00
0.74**
-0.07ns

1.00
0.40**

1.00

GYPH

0.31*

0.39**

0.59**

0.78**

0.35*

PAD= Spad reading, NKRPE=number of kernel row/ear, NKPR=number of kernel/row, EW=

Ear Weight, FHKW= Five hundred kernel weights, GYPH= grain yield/ha

GYPH

1.00

143

Figure1: Meteorological data during the growing period of Maize at Rampur Chitwan 2014/15
Chart of GYPH vs GENOTYPES
2500

2376

2222

GYPH (Kg)

2000

2175

2232
1982

1922

1598

1500

1278
1073

1000
728

500

820

711

493
164

)
1
L
Y
1
N
k)
46
15
17
27
15
EV L -17
/L
40
C A YB( 3
ET
YC
ch
-4
-1
ololol ol-2
EY
RI
LO
EE
A
td
O P RUN POO
Po
po
po
po
3T
(s
VA
SS
GH
/P
/
1/
Y/
AR
O
A
5
2
O
Y
S
2
K
C
H
S
E
R
A
44
R.
-6
ER
UN
7T
A C RA J
PZM
RM
S9
AR
PO
FA
GENOTYPES

Figure 2: Genotypes with their yields (kg/ha)

Figure 3. Estimated liner correlation (a) between SPAD reading and grain yield/ha (GYPH), (b) between NKRE (number of kernel row/ear) and
grain yield/ha (GYPH), (c) between NKPR (number of kernel/row) and grain yield/ha (GYPH), (d) between EW (ear weight) and grain yield/ha
(GYPH), (e) between FHKW (five hundred kernel weight) and grain yield/ha (GYPH)

144
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