RESEARCH ARTICLE

%
SABRAO Journal
of Breeding and Genetics
48 (4) 518-527, 2016

COMPARATIVE ANALYSIS OF GENETIC DIVERSITY OF MAIZE INBRED LINES

FROM KASHMIR VALLEY USING AGRO-MORPHOLOGICAL AND SSR MARKERS
M. MUSHTAQ1, M.A. BHAT2, J.A. BHAT1*, S. MUKHTAR1 and A.A. SHAH3
1

School for Biotechnology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, Chatha,
Jammu-180009Jammu & Kashmir, India
2
Division of Plant Breeding and Genetics, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir,
Shalimar, Srinagar, Jammu & Kashmir-190025, India
3
Division of Plant Breeding and Genetics, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu,
Jammu-180009, India
*Corresponding authors email: [email protected]
Email addresses of co-authors: [email protected], [email protected], [email protected],
[email protected]

SUMMARY
In India maize is emerging as third most important crop that contributes 2.5 billion dollar to Indian agriculture GDP.
The maize productivity figures are low primarily because of the cultivation of landraces and composite varieties.
Therefore, efforts are needed to develop hybrids for exploiting maximum heterosis for increased yield and quality
traits. In this regard, 20 maize inbred lines suited to high altitude and plain areas of Kashmir Valley are
comparatively evaluated for diversity analysis using both agro-morphological and SSR markers. Analysis of
variance showed that all the characters except ear girth were significantly different (P < 0.01) among the genotypes.
The first four principal components (PCs) of the PCA analysis contributed 97.9% of the variability. The dendrogram
obtained through agro-morphological and SSR analysis separated the genotypes into three (I, II and III) and four (I,
II, III and IV) major clusters, respectively. Out of 25 SSR markers tested only 10 primer pairs were found
polymorphic and detected a total of 31 alleles with an average of 3.1 alleles per locus. The maximum and minimum
polymorphic information content (PIC) values were found to be 0.78 and 0.29 for the primers Phi022 and
Phi109188, respectively. The Mantel test revealed a non-significant low correlation (r = 0.12, P < 0.148) between
the agro-morphological and SSR matrices. Both methods result in diverse clustering of genotypes suggesting
considerable diversity among the studied genotypes. The diverse maize inbred lines identified can be used as parents
in exploiting heterosis as well as to identify transgressive segregants for yield and quality traits of maize. Hence,
both methods proved effective for the diversity analysis of maize inbreds, and their combined study provides useful
information.

Keywords: Genetic diversity, maize inbreds, agro-morphological, SSR, dendrogram

Key findings: The combined genetic diversity analysis using both agro-morphological and SSR markers
provides useful information and identified diverse maize inbred lines which could be effectively used as
parents in heterosis and transgressive breeding.
Manuscript received: June 23, 2016; Decision on manuscript: October 19, 2016; Manuscript accepted: November 2, 2016.
Society for the Advancement of Breeding Research in Asia and Oceania (SABRAO) 2016
Communicating Editor: Naqib Ullah Khan

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SABRAO J. Breed. Genet. 48 (4) 518-527

nature. These markers have been used for

studying the genetic relationship in maize
(Adeyemo et al., 2011; Ristic et al., 2013;
Sserumaga et al., 2014; Salami et al., 2016), rice
(Kunusothet al., 2015), wheat (Arora et al.,
2014), and many other crops including pea
(Handerson et al., 2014), faba bean (Abid et al.,
2015), and soybean (Bisen et al., 2015).
The combined analysis
through
morphological and molecular marker is the best
option to characterize inbred lines giving an
opportunity to comparatively analyze the
phenotypes from field experiments with
molecular phenotypes and genotypes from
laboratory studies. Comparison of different
methods in genetic studies provides researchers
and plant breeders with more information in the
screening and selection process. Keeping this in
view, this study is undertaken to estimate the
genetic distance of 20 maize inbred lines
collected from Kashmir Valley of State of
Jammu and Kashmir using a combination of
morphological and SSR markers, which will
allow us to study the relative efficacy of these
two methods in cultivar differentiation as well as
identification of diverse inbred lines that can be
used as parents in exploiting heterosis for
maximizing maize production and productivity.

INTRODUCTION
Maize (Zea mays L.) also known as corn, is the
only cereal crop of American origin that is
cultivated in tropical and subtropical regions
throughout the world. Maize is currently
produced on nearly 100 million hectares in 125
developing countries and is among the three
most widely grown crops in 75 countries (FAO
STAT, 2010). In India, maize is emerging as
third most important crop after rice and wheat,
and contributes 2.5 billion dollar to Indian
agriculture GDP (Kumar et al., 2013). India
occupies fifth place in average under maize in
the world after US, Brazil, China and Mexico.
As regards, Jammu & Kashmir it plays an
important role in the livelihood of the people of
this hilly and sub-mountainous state and
occupies highest area in the state, but
productivity figures are very low primarily
because of the cultivation of landraces and
composite varieties. Therefore, efforts are
required to develop hybrids for exploiting
maximum heterosis in order to increase
production and productivity of maize.
The choice of parents is the initial step
in plant breeding and directly benefit
transgressive segregation and heterosis, which is
considered to be high by the parents that are
distantly related (Joshi et al., 2004). The diverse
parents are observed to give progeny with higher
heterosis (Joshi and Dhawan, 1966; Anand and
Murrty, 1968). Thus, genetic diversity
estimation of crop species determines its
potential for improved efficiency and its use for
breeding, which inevitably prompted increased
food production. The genetic diversity among
individuals/populations can be determined using
different markers systems viz., morphological,
biochemical and molecular.
Morphological traits have been already
used to assess genetic diversity of maize
genotypes by number of earlier studies (Kashiani
et al., 2014; Azad et al., 2012; Syafii et al.,
2015; Kumar et al, 2015). Various types of
molecular markers (RFLP, RAPD, AFLP, ISSR
and SSR etc.) are available for varietal
identification and genetic characterization of
crop germplasm, among them SSR are the
marker of choice being co-dominant, multiple
allelic, simple, reproducible and reliable in

MATERIALS AND METHODS

Plant material and experimental site
A set of 20 inbred lines of maize suited to high
altitude and plain areas of Kashmir valley were
selected for genetic divergence studies. These
lines comprised of promising inbred lines from
Dryland (Karewa) Agriculture Research Station,
Budgam and High Altitude Maize Research,
Sub-Station, Sagam, Anantnag. The genotypes
included lines from CIMMYT, Mexico. All the
lines were in the advanced stage of development.
Details about the origin and pedigree of these
genotypes are provided in Table 1. Some of the
lines used in this study are also currently used as
parental lines of two popular maize hybrids
released in Kashmir. W3 and W5 are the parents
of the first single cross hybrid viz., Shalimar
Maize Hybrid-I, whereas KDM-500 is the male
parent of Shalimar Maize Hybrid-II, which has

519

Mushtaq et al. (2016)

been proposed for release in recently held 31st

ZREAC meeting.
The experimental material for the
present research was laid out at the experimental
area of the Dryland (Karewa) Agriculture
Research Station, Budgam and Centre for Plant
Biotechnology, Sher-e-Kashmir University of
Agricultural Sciences and Technology of
Kashmir, Shalimar, Srinagar, J & K.

DNA extraction and SSR analysis

Total genomic DNA was isolated from young
leaves at 5 leaf stage from 8-10 field grown
plants of each inbred line (approximately 5-7g of
fresh weight) using CTAB (CetylTrimethyl
Ammonium Bromide) method as modified by
Saghai-Maroof et al. (1984). Quantification of
DNA samples was done by using Nanodrop
(mySPEC, Scientific GmbH, Germany) and
quality was estimated by using 0.8% agarose gel
electrophoresis. High concentration of DNA
samples was further diluted in 10:1 Tris-EDTA
to a working concentration of 50ng/l andstored
at 4C for PCR based marker analysis. A total of
25
pairs
of
SSR
primers
flanking
themicrosatellite region previously developed
and published by Sharopova et al. (2002)
wereselected. After testing the 25 primers, a
total of 10 primers were found polymorphic and
used for further analysis. Detailed description of
the primers is available at Maize DB:
http://www.agron.missouri.edu;
http://www.gramene.org/markers/microsat/.
PCR reaction was prepared with 50 ng of rice
genomic DNA, 0.2 g of 3 and 5 end primers,

Morphological characterization
The data on ten agro-morphological traits are
recorded from five randomly selected
representative plants in all the genotypes in each
replication. The standard method of DUS test
(Distinctiveness, Uniformity and Stability, Govt.
of India) was used for recording observation for
each of the character which includes plant
height, ear height, days to 50% pollen shed, days
to 50% silking, ear length, ear girth, 100 grain
weight, grain yield per plant, grain yield and
shelling
%
(Biodiversity
protocols:
www.bioversity.org). The mean values of the
data obtained were used for the various
statistical analyses.

Table 1. List of the maize (Zea mays L.) genotypes used for morpho-molecular characterization with their
name, source population, altitude and place of collection.
Line
KDM-361A
KDM-343A
KDM-332A
KDM-914A
KDM-895A
KDM-340A
KDM-362A
KDM-916A
KDM-500A
CM-502
W5
462
30
53
401
W3
YI-1
114-2
460
39

Source population
F-7012
Seed Tech 3435
AAMH 204
AH-1139
C-170
PRO-349
DMR-6520
DMR-00RIBK114
CM-128
CM-502
CML-354
CML-349
-

Altitude
Low and mid
Low and mid
Low and mid
Low and mid
Low and mid
Low and mid
Low and mid
Low and mid
Low and mid
Low and mid
High
High
High
High
High
High
High
High
High
High

Place of collection
Dryland (Karewa) Agriculture Research Station, Budgam
Dryland (Karewa) Agriculture Research Station, Budgam
Dryland (Karewa) Agriculture Research Station, Budgam
Dryland (Karewa) Agriculture Research Station, Budgam
Dryland (Karewa) Agriculture Research Station, Budgam
Dryland (Karewa) Agriculture Research Station, Budgam
Dryland (Karewa) Agriculture Research Station, Budgam
Dryland (Karewa) Agriculture Research Station, Budgam
Dryland (Karewa) Agriculture Research Station, Budgam
Dryland (Karewa) Agriculture Research Station, Budgam
High Altitude Maize Research, Sub-Station, Sagam, Anantnag
High Altitude Maize Research, Sub-Station, Sagam, Anantnag
High Altitude Maize Research, Sub-Station, Sagam, Anantnag
High Altitude Maize Research, Sub-Station, Sagam, Anantnag
High Altitude Maize Research, Sub-Station, Sagam, Anantnag
High Altitude Maize Research, Sub-Station, Sagam, Anantnag
High Altitude Maize Research, Sub-Station, Sagam, Anantnag
High Altitude Maize Research, Sub-Station, Sagam, Anantnag
High Altitude Maize Research, Sub-Station, Sagam, Anantnag
High Altitude Maize Research, Sub-Station, Sagam, Anantnag

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SABRAO J. Breed. Genet. 48 (4) 518-527

200 mM of each dNTP, 1X PCR buffer

containing 50 mM KCL, 10 mMTrisHCl
(pH8.9), 2.0 mM MgCl2 and one unit of Taq
Polymerasein a total of 25 L solution
individually for all 14 primer pairs. PCR thermal
cycler was programmed for 1 min at 94C, 1 min
and 30 seconds at 55C, 1 min at 72C and a
final cycle of 10 min at 72C.Amplification
product was separated on 3.5% of agarose gel in
1X TBE buffer followed by staining with
ethidium bromide.

arithmetic averages (UPGMA) and sequential,

agglomerative, hierarchical and nested (SAHN)
clustering algorithm to obtain a dendrogram.
Comparison
between
agromorphological and SSR data was performed by
calculating the correlation between the agromorphological and SSR similarity matrices
through mantel test (Mantel, 1967) with 1000
permutations using PASSaGE 2 software.

RESULTS
Data analysis
Agro-morphological analysis
Analysis of variance was performed for all agromorphological traits in order to test the
significance of variation among the genotypes
using SPSS 16.0 software. Cluster analysis was
done to yield a dendrogram depicting the
morphological relatedness of the inbred lines.
Principal component analysis (PCA) was also
used to detect underlying sources of
morphological variability, and to investigate
patterns of genetic diversity (Mohammadi and
Prasanna, 2003). Bray-Curtis distances and
UPGMA (Unweighted pair group method with
arithmetic mean) was the clustering method and
all these analyses were done using the PAST
software (Hammer et al., 2001).
For SSR data, the presence or absence of
the band was scored as 1 or 0, respectively. In
order to determine the utility of the SSR
markers, Number of alleles per marker,
Polymorphic Information Content (PIC),
Effective multiplex ratio (EMR) and Marker
Index (MI) were calculated. The Polymorphism
Information Content (PIC) values of individual
primers were calculated based on the formula
PIC= 1- ni=1 P2ij (Anderson et al., 1997).
Marker Index, a product of information content,
as measured by PIC, and Effective Multiplex
Ratio (EMR), was calculated following (Powell
et al., 1996). The Jaccards similarity index was
calculated using NTSYS-pc version 2.02e
(Applied Bio-Statistics, Inc., Setauket, NY,
USA) package to compute pair wise Jaccards
similarity coefficients (Jaccard, 1980) and this
similarity matrix was used in cluster analysis
using an unweighted pair-group method with

The analysis of variance showed that mean

squares due to genotypes were highly significant
(P 0.01) for all characters except ear girth
which showed non-significant variation (Table
2). Principle component analysis (PCA) of the
agro-morphological traits showed that the first
four principal components together accounted
for 97.929% of the total phenotypic variation
(Table 3). The first principal component (PC1)
accounted the maximum portion of 83.364% of
total variance, and characters that contribute
more positively to this component were plant
height, ear height, days to 50% silking, ear
length, grain yield per plant, grain yield and
shelling%. The second component (PC2), which
featured ear height as the principal trait,
explained an additional 8.276% of the
phenotypic variation. Finally, third and fourth
principal component (PC3 and PC4) contributed
around 3.762% and 2.527%, respectively of the
variability present among the accessions for the
traits used in this study. The PC3 explained the
pattern of variation in 100 grain weight, and for
PC4 the maximum variation is contributed by
days to 50% pollen shed. The dendrogram
obtained using phenotypic characters separated
the genotypes into 3 major clusters (I, II and III)
consisting of 8, 2 and 10 genotypes, respectively
with Bray-Curtis distance ranging from 0.864 to
0.992 (Figure 1). The cluster I and II comprised
of inbred lines from low and mid altitudes, while
cluster III consists of all ten inbred lines from
high altitude.

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Mushtaq et al. (2016)

Table 2. Analysis of variance for yield and yield component traits in maize (Zea mays L.).
Mean squares
Source
of variation
Replication
Treatment
Error

d.
f.

Plant
height
(cm)

Days to
50%
pollen
shed

Ear
girth
(cm)

Ear
length
(cm)

100
Grain
weight
(g)

119.75**

38.57**

41.75**

46.51**

0.38*

4.57**

0.01NS

19

93.34**

222.59**

311.25**

318.66**

0.32NS

15.26**

19

0.63

3.18

0.34

0.35

0.07

0.11

Days to
50%
silking

Ear
height
(cm)

Grain
yield
(q ha1
)

Shellin
g %age

39.64**

8.44**

27.22**

23.77**

105.04**

69.37**

150.75**

0.02

0.14

0.49

0.96

Grain
yield
plant-1

Table 3. Eigenvectors, Eigen values, total and cumulative variability (%) for 20 maize genotypes based
on ten agro-morphological traits.
Principal component (axes)
Eigen value
Variability (%)
Cumulative (%)
Traits
Plant height (cm)
Ear height (cm)
Days to 50% pollen shed
Days to 50% silking
Ear length (cm)
Ear girth (cm)
100 Grain weight (g)
Grain yield per plant (g)
Grain yield (q ha-1)
Shelling %

PC1
8.336
83.364
83.364
0.322
0.334
0.3233
0.342
0.332
0.169
0.283
0.341
0.339
0.336

PC2
0.827
8.276
91.64

PC3
0.376
3.762
95.402
Eigenvectors
0.008
0.091
-0.117
0.075
0.118
-0.347
-0.049
-0.081
-0.093
-0.228
0.949
0.216
-0.233
0.848
-0.056
-0.143
-0.049
-0.072
-0.035
-0.125

PC4
0.253
2.527
97.929
-0.699
0.303
0.478
0.176
-0.322
-0.004
0.169
-0.054
-0.127
0.092

Figure 1. Dendrogram based on agro-morphological traits showing three clusters (I, II and III) of 20
maize genotypes using UPGAMA method and Bray-Curtis distance.

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SABRAO J. Breed. Genet. 48 (4) 518-527

Figure 2. Gel picture showing banding pattern of 20 maize genotypes with phi022 marker.

clusters I, II, III and IV comprising of 6, 7, 4 and

3, respectively with Jaccards similarity
coefficient ranging from 0.15 to 0.95. Cluster IV
consist of 2 inbred lines from high altitude and
one inbred line from low and mid altitude, while
as cluster II consist of 4 genotypes from high
altitude and 3 from low and mid altitude. The
cluster III comprises mostly inbred lines from
high altitude except inbred line 10 from low
altitude, whereas cluster I consist of all inbred
lines from low and mid altitude except inbred
line 17 from high altitude (Figure 3).

Molecular marker analysis

All the 20 maize inbred lines were genotyped
with 10 polymorphic SSR markers; and are
selected for their ability to produce amplified
product
at
optimum
concentration,
polymorphism level among the genotypes and
consistency of the pattern. Total 31 alleles were
scored from these primer pairs, and 100 percent
were found polymorphic. The gel picture
showing a banding pattern of 20 maize inbred
lines with phi022 marker is presented in Figure
2. The respective values of overall genetic
variability for Polymorphism Information
Content (PIC), Effective multiplex ratio (EMR),
Number of alleles per locus and Marker Index
(MI) across all the 20 genotypes are given in
Table 4. Highest PIC value (0.78) was observed
for the primer Phi022 and lowest PIC value
(0.29) was recorded for the primer Phi109188
(Table 4), with an average 0.58. The MI values
ranged from 1.19 to 0.01 with an average of
1.00. The EMR is a feature of marker that
indicates the discriminatory potential of the
primer, and ranged from 2.57 to 0.05 with an
average of 1.06. The allele number per locus
varied from 2 to 4, with an average of 3.1 alleles
per locus(Table 4). The SSR data were also
subjected to genetic cluster analysis to further
elucidate the relationship among the genotypes
and the dendrogram generated through UPGMA
analysis have been presented in Figure3, which
grouped all maize inbred lines into 4 major

Comparison of agro-morphological and SSR

markers
The mantel test showed non-significant low
correlation (r = 0.12, P < 0.148) between the
agro-morphological and SSR data. Both agromorphological and molecular analysis allowed
separation of advanced maize inbred lines into
different clusters of 3 and 4, respectively. The
two methods showed considerable discrepancies
between dendrograms as far as the grouping of
genotypes is considered. For instance, the inbred
lines from high altitude which are morphological
clustered in cluster III were grouped into 4
separate clusters (I, II, III and IV) in SSR
analysis (Figures 2 and 3). The range of agromorphological data based genetic distance
between pairs of genotypes was considerable
narrow (0.864 to 0.992) as compared to that of
SSR based data (0.15-0.95).

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Mushtaq et al. (2016)

Table 4. List of markers used, chromosome number, number of alleles, PIC value, effective multiplex
ratio (EMR) and marker index (MI).
Markers
Phi034
Phi022
Phi015
Phi006
Zcaa391
Phi101049
Phi109188
Phi063
Phi064
Phi053
Average

Chromosome
Number
7
3
4
4
9
10
5
10
1
3

Number of
alleles
2
3
4
4
4
3
2
3
3
3
3.1

PIC
0.48
0.78
0.58
0.71
0.56
0.62
0.29
0.43
0.77
0.59
0.58

Effective multiplex ratio

(EMR)
0.47
2.57
0.84
1.92
1.38
0.88
0.05
0.20
1.31
0.94
1.06

Marker index
(MI)
0.97
1.19
1.07
1.10
1.01
1.19
0.01
0.85
1.09
1.11
1.00

Figure 3. UPGMA dendrogram based on SSR data showing four clusters (I, II, III and IV) of 20 maize
genotypes.

breeding remains the choice of methods

considering its success over years. A logical way
to start any breeding programme is to survey the
variation present in the available germplam
resources. In this regard, 20 maize inbred lines
collected from low, mid and high altitudes of
Kashmir Valley are characterized using both
agro-morphological and molecular markers. The
aim of our study was to identify the divergent
inbred lines that will be subsequently used in
maize breeding to exploit heterosis for yield and
other quality traits of maize. The genetic

DISCUSSION
The analysis of genetic diversity and relationship
among the elite breeding materials can
significantly aid in crop improvement (Hallauer
et al., 1988). In maize, this information is useful
in planning for hybrid and line development,
assigning lines to heterotic groups and in plant
variety protection (Yuan et al., 2002). There
exists an urgent need to promote maize breeding
to meet the increasing demands for maize grain
and its products. In this context, maize hybrid

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SABRAO J. Breed. Genet. 48 (4) 518-527

diversity estimates are often biased by the choice

of data viz., phenotypic and molecular marker.
Therefore, in the current study both types of data
have been used to measure unbiased diversity
estimation. In this study, all the agromorphological traits except ear girth revealed
significant ( 0.01) variations indicating the
presence of sufficient amount of genetic
variability among the maize inbred lines for all
the traits. In maize significant variations was
also reported earlier by other researchers for
various morphological traits (Saleem et al.,
2002; Ishaq et al., 2015; Hussain et al., 2014).
PCA is an effective technique giving
information about traits that are more important
for the breeder to conduct specific breeding
programs Salimi et al. (2012). In our study, first
four PCs explained 97.929% of variation among
20 maize inbred lines and these results were
supported by the finding of Ristic et al. (2013);
Azad et al. (2012), who studied maize genotypes
of Serbia and Bangladesh, respectively. The
morphological cluster analysis divided the 20
maize inbred lines into 3 major 4 clusters with
all the inbred lines from low and mid altitude
clustered into cluster I and II, and inbred from
high altitude into separate cluster III. The
morphological clustering of breeding lines is to
some extent in good agreement with their place
of collection and altitude.
In this study, a total of 31 alleles were
detected by 10 polymorphic SSR markers among
20 maize inbred lines with an average number of
3.1 alleles per locus and average PIC value of
0.58, which was also observed in maize by
earlier studies (Adeyemo et al., 2011;
Sserumaga et al., 2014). The UPGAMA analysis
based on SSR data divided the genotypes into 4
major clusters I, II, III and IV. The inbred lines
from plain areas (low and mid altitude) and high
altitude do not form separate clusters as in
morphological analysis but are clustered
together in all the 4 major clusters. It is because
SSR provides
more information than
morphological analysis distinguishing some
genotypes that are not morphologically
distinguished.
The Mantel test revealed a nonsignificant low correlation between the agromorphological and SSR matrices (r = 0.12, P <
0.148) of the 20 maize inbred lines, showing that

these methods discriminated very differently

among the genotypes. As the two methods are
based on different criteria, agro-morphological
dendrogram are constructed based on phenotypic
data, Bray-Curtis distances and UPGMA,
whereas molecular phylogenetic tree is based on
SSR data, Jaccards similarity coefficient and
UPGMA.
Low
correlation
between
morphological and molecular markers has been
reported in many crops (Koehler-Santos et al.,
2003; Ferriol et al., 2004; Bushehri et al., 2005)
and these authors suggest that it could be as a
result of the independent nature of
morphological and molecular variations. The
other reason may be that a large portion of
variation detected by molecular markers is nonadaptive and is therefore not subject to either
natural or artificial selection as compared with
phenotypic characters, which in addition to
selection pressure are influenced by the
environment (Vieira et al., 2007).
In conclusion, this study showed that
both morphological and molecular analysis
results in the diverse clustering of 20 maize
inbred lines with the latter showed more
diversity among the studied genotypes as
compared to morphological analysis as indicated
by similarity coefficient distribution. Therefore,
it is evident that the studied maize inbred lines
from Kashmir Valley possess fair amount of
genetic diversity, and the diverse lines identified
can be effectively utilized as parents in
exploiting heterosis as well as to identify
transgressivesegregants for higher yield and
quality traits of maize. This will lead to
increased maize production and income of
resource poor farmers of state Jammu &
Kashmir to make them self-sufficient. In
addition, these inbred can be potentially used as
donors in breeding programme as well as in the
development of bi-parental and multi-parent
mapping populations for identification of
genes/QTLs for yield, yield contributing and
quality traits of maize. Hence both agromorphological and SSR markers proved to be
useful in genetic diversity analysis of maize
inbreds.

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Mushtaq et al. (2016)

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ACKNOWLEDGEMENTS
We thank the Vice Chancellor and Centre for Plant
Biotechnology of SKUAST-K for providing the lab
facilities and financial assistance for carrying this work.
Conflict of Interest: The authors declare that they have no
conflict of interest.

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