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Assessing drought tolerance using PEG-6000 and molecular screening by SSR

markers in maize (Zea mays L.) hybrids

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Maydica
Original paper

Assessing drought tolerance using PEG-

Open Access

6000 and molecular screening by SSR

markers in maize (Zea mays L.) hybrids
R. Nirmal Raj, J. Gokulakrishnan and M. Prakash*

Department of Genetics and Plant Breeding, Annamalai University, Annamalainagar-608 002, Tamilnadu, India
* Corresponding author : M. Prakash, Professor, Department of Genetics and Plant Breeding, Annamalai University, Annamalainagar-608 002,
Tamilnadu, India
E-mail: geeth_prakash@yahoo.co.in
KeyWords Drought, PEG-6000, Osmotic stress, SSR marker, Nei’s matrix, Genetic identity

Abstract
Maize is a versatile crop, widely grown across the world but its yield potential is severely affected by drought
stress due to frequent monsoon failure. Identification of tolerant maize hybrids at seedling stage itself could be
a better option than evaluation at maturity stage. Hence, experiments were conducted to screen maize hybrids
under induced PEG stress and also with molecular marker analysis. A laboratory experiment was conducted with
twenty one hybrids and the experiment was laid out under completely randomized design (CRD) with two repli-
cations. PEG-6000 treatment was applied in three concentrations (0, 10 and 20%) which induced osmotic stress
levels of 0, -3 and -6 bars respectively. The experimental results revealed that there was significant reduction in
seedling characteristics viz., germination percentage, shoot length, seminal root length, fresh weight, dry weight,
seed vigour index I and seed vigour index II with increase in PEG concentration. The per se performance revea-
led that the hybrids AUK-30 and AUMH-8855 were drought tolerant. Promptness index and germination stress
tolerance index were found as reliable indicators to screen drought tolerant hybrids at seedling stage. Fourteen
drought linked SSR markers were used to characterize the hybrids of which the primer umc1962 was highly infor-
mative in identifying tolerant hybrids as it formed distinct bands. The earlier identified tolerant hybrids showed
highest genetic identity values in Nei’s matrix. Hence, the hybrids AUK-30 and AUMH-8855 resulted drought
tolerant both phenotypically and genotypically

Introduction developmental stages such as, germination, shoot

Maize (Zea mays L.) is the third most important cereal
crop after wheat and rice (Cooper et al., 2014). It is a
multi faceted crop which is mainly used for food and
forage. Considering its significance, there is a growing
focus on the selection of maize germplasm to evaluate
its drought or water stress tolerance (Avramova et al.,
2015).
Drought is one of the most serious world-wide
problems for agriculture, which is mostly attributed to
monsoon failure and it can be defined as the absence
of soil water to provide conditions for crops to grow
as a consequence of precipitation being less than
normal (UNISDR, 2009). Four tenths of the world’s
agricultural land lies in arid or semi arid regions and
maize is cultivated mainly in dry land (69%) rather
than in irrigated fields (21%) (Efendi, 2009). Transient
droughts can cause death of livestock, famine and social
dislocation. As water resources for agronomic uses
become more limiting, the development of drought
tolerant lines become increasingly more important.
Water stress has damaging effects at
growth, root growth and flowering (Rauf et al., 2007;
Khayatnezhad et al., 2010). Water stress adversely
affects seed emergence and also increases the
mean germination time (Mostafavi et al., 2011;
Khodarahmpour, 2011). The drought induced by PEG is
due to osmotic stress resulting from its higher molecular
weight than any other similar compunds (Sidari et al.,
2008; Partheeban et al., 2017).
Conventional breeding for drought resistance is time
consuming and is greatly constrained by inadequate
screening methods, as well as lack of knowledge
of the genetics of drought resistance (Bawa et al.,
2015). There is also lack of alternative and reliable
screening assays that can adequately predict the field
performance of a given genotype (Omanya et al., 2001).
To overcome such barriers, efforts should be made
to identify molecular markers such as SSRs and SNPs
associated with yield and tolerance traits (Maheswari
et al., 2016). Molecular approach accelerates the
genetic enhancement of drought tolerance in maize
(Nguyen et al., 2012; Thirunavukkarasu et al., 2014;
various Beyene et al., 2015). These markers will not only ease

64 ~ M 19 Maydica electronic publication - 2019

 Drought screening in maize hybrids 2

the screening for drought tolerance, but also help in
tracking important drought resistance genes.
From this perspective, the present study was carried out
to identify drought tolerant maize hybrids using PEG
6000 and to characterize maize hybrids at molecular
level
Materials and Methods
Genetic materials
Stress tolerance indices and drought traits
measurements
Observations were recorded on each day up to
seven days. After seven days, emergence percentage,
shoot length, root length, seminal root length, dry
weight, fresh weight and other drought indices were
recorded. Stress tolerance indices were calculated
following the protocol given by International Seed
Testing Agency (ISTA 1996) and other authors.

The present investigation on drought screening was EP (%) = (NGS / TNS) × 100
conducted in Abiotic Stress Laboratory, Department of
Where, EP is Emergence percentage, NGS is the
Genetics and Plant Breeding, Faculty of Agriculture,
Number of Germinated Seeds and TNS is the Total
Annamalai University. Twenty one hybrids of single
Number of viable Seeds taken for the experiment
cross origin were used to study the effect of drought
(Scott et al., 1984).
stress (Table S1).

Table 1 - Mean comparison of main effects of drought stress levels on drought related traits

Drought Stress EP (%) PI (%) GSTI (%) SL (cm) SLSI (%) RL (cm) RLSI (%) SRL (cm) FW (gm) DW (gm) SV I (%) SV II (%)

Control 96.67 9.54 100.00 3.58 100.00 4.26 100.00 4.39 0.48 0.24 758.52 23.39
T1 (10%) 46.90 4.33 45.23 1.66 44.56 5.19 122.69 3.71 0.43 0.22 353.97 10.60
T2 (20%) 14.29 1.24 12.84 0.20 5.74 1.91 43.93 0.67 0.22 0.12 64.49 3.29

EP- Emergence percentage, PI- Promptness index, GSTI- Germination stress tolerance index, SL- Shoot length, SLSI- Shoot length stress index, RL-
Root length, RLSI- Root length stress index, SRL- Seminal root length, FW- Fresh weight, DW- Dry weight, SV I- Seed vigour I, SV II- Seed vigour II

PI (%) = nd2 (1.00) +nd4 (0.75) +nd6 (0.5) +nd8 (0.25)

PEG-6000 Screening for drought tolerance
A drought stress screening was conducted by using
PEG-6000 on germination and early seedling growth
characters and experiments were laid out in completely
randomized design. The osmotic stress tolerance study
was performed in petri plates with filter paper. The
seeds were selected for size homogeneity, surface
sterilized for 5 min in 1% (v/v) sodium hypochlorite
and then rinsed twice in distilled water. Ten seeds
of each hybrid were placed in the petri plates with
corresponding PEG concentration (0, 10 and 20%). The
solutions were applied on a daily basis after draining
the previous day solution. The osmotic stress induced
(0, -3 and -6 bar) by each concentration was calculated
by using the equation of Michel and Merril (1973) at an
average temperature of 33°C.
Ψs = –(1.18 x 10–2) C – (1.18 x 10–4) C2 + (2.67 x 10–4) CT
+ (8.39 x 10–7) C2T
Where, Ψs = Osmotic potential (bar)
C = Concentration (g L–1 PEG-6000 in water)
T = Temperature (°C)
Where, PI is Promptness Index, nd is the number of
seeds germinated on the day of observation (George,
1967).
GSTI (%) = [PI of stressed seeds / PI control seeds] x 100
Where, GSTI is Germination Stress Tolerance Index.
SLSI (%) = (Shoot length of Stressed plant / Shoot
length of control plants) x 100
Where, SLSI is Shoot length stress index
RLSI (%) = (Root length of stressed plant / Root length
of control plants) x 100
Where, RLSI is Root Length Stress Index.
Seed vigour I (SVI) = germination percentage ×
seedling length
(Abdul Baki and Anderson, 1973).
Seed vigour II (SVII) = germination percentage × dry
weight (Abdul Baki and Anderson, 1973).

64 ~ M 19 Maydica electronic publication - 2019

 Drought screening in maize hybrids 3

Statistical analyses amplified DNA products were analyzed on an agarose

The data recorded on different traits were subjected
to analysis of variance, combined analysis of variance,
per se performance and correlation using TNAUSTAT
software’s and PB tools
Molecular Marker analysis (SSR)
Leaf sheath samples were collected from two weeks
old maize seedlings and stored at minus 20°C until
used for DNA extraction.
DNA was isolated by using ORIGIN plant DNA
gel (3%) and products were visualized under ultraviolet
light after ethidium bromide staining (8 µl). PCR
products were resolved through electrophoresis carried
out at 110 V. The DNA fragments were visualized and
documented in gel documentation system.
The separated bands were scored based on the
reference product size and the allelic variations were
scored as A, B and C respectively. These scored
products were analyzed using POPGENE ver. 3.2
software to find out the genetic distance and diversity
between the maize hybrids

Table 2 - Mean comparison of main effects of maize hybrids on drought related traits

EP PI GSTI SL SLSI RL RLSI SRL FW DW SV I SV II

Genotypes
(%) (%) (%) (cm) (%) (cm) (%) (cm) (gm) (gm) (%) (%)
G1 45.00 4.08 45.37 2.11 52.73 3.60 74.85 2.39 0.30 0.16 390.35 10.48
G2 53.33 5.05 58.52 2.92 55.45 3.08 81.63 1.92 0.37 0.19 404.83 10.67
G3 46.67 4.57 45.70 2.92 53.11 4.38 62.53 3.24 0.32 0.14 541.93 9.83
G4 58.33 5.61 56.80 1.77 63.15 5.07 88.88 3.43 0.39 0.21 453.03 13.18
G5 86.67 8.40 84.02 2.77 65.26 6.73 112.15 4.90 0.43 0.25 849.08 21.08
G6 43.33 4.19 43.25 0.90 37.45 0.94 70.95 1.42 0.19 0.08 141.73 5.22
G7 43.33 4.17 46.33 1.61 50.08 3.25 69.00 2.88 0.28 0.13 326.12 9.17
G8 46.67 4.54 45.43 1.24 43.95 3.02 75.12 3.53 0.24 0.11 307.60 7.73
G9 76.67 7.46 74.63 2.26 57.23 5.51 129.84 4.94 0.44 0.25 618.00 18.93
G10 46.67 4.72 47.73 1.95 47.50 4.70 86.54 2.01 0.50 0.31 390.83 16.00
G11 40.00 4.27 42.72 1.49 38.46 3.31 71.77 1.04 0.33 0.19 313.30 11.90
G12 50.00 5.01 50.05 2.68 47.16 3.69 100.82 2.94 0.47 0.19 394.45 9.70
G13 46.67 4.47 46.35 1.25 37.36 1.83 72.20 1.26 0.20 0.09 240.73 6.53
G14 53.33 5.17 52.36 1.29 52.71 3.43 106.86 2.38 0.48 0.23 289.15 12.90
G15 50.00 4.30 53.68 0.92 51.30 3.48 111.72 3.17 0.44 0.22 240.32 11.60
G16 40.00 3.70 39.15 1.32 46.03 3.06 76.22 2.93 0.42 0.24 271.40 15.27
G17 41.67 3.99 46.13 1.14 49.11 1.74 95.28 2.19 0.41 0.26 155.15 10.42
G18 60.00 5.68 58.08 1.86 53.94 5.16 103.00 4.80 0.39 0.21 472.48 12.05
G19 71.67 6.59 67.54 2.83 56.37 5.65 113.89 3.75 0.48 0.27 675.07 18.67
G20 46.67 4.32 46.02 0.99 40.89 2.57 59.74 2.02 0.44 0.26 277.13 18.03
G21 58.33 5.46 56.62 1.96 52.90 5.33 103.36 4.38 0.38 0.19 486.17 11.58

EP- Emergence percentage, PI- Promptness index, GSTI- Germination stress tolerance index, SL- Shoot length, SLSI- Shoot length stress index, RL-
Root length, RLSI- Root length stress index, SRL- Seminal root length, FW- Fresh weight, DW- Dry weight, SV I- Seed vigour I, SV II- Seed vigour II

purification kit protocol. The integrity of total

DNA isolated is assessed through agarose gel
electrophoresis (0.8% agarose gel containing 0.8 µg/ml
ethidium bromide). Fifteen SSR primer pairs which were
reported for association with traits related to drought
tolerance belonging to different series viz., bnlg, umc,
phi were selected and annealing temperatures (TA)
were standardized (Table S2). The PCR assay mixture
contained 15 picomol of each primer pairs, 200 µM of
deoxy nucleotides, 2.5 µl of 1X buffer with MgCl2, 1
unit Taq polymerase and 25-50 ng of DNA template.
The PCR reaction was carried out at 94°C for 2 min,
followed by 35 cycles of 94°C for 30 sec, specific
annealing temperature for 1 min, after 2 min extension
at 72°C and a final extension step of 7 min at 72°C. The
Results and discussion
Genotypic and phenotypic drought tolerance
variance
The combined analysis of variance showed significant
difference between hybrids under drought stress and
revealed significant treatment effect on hybrids for
all the twelve characters taken for study and hence
selection of a drought tolerant hybrid was possible
(Table S3).
The mean performance of genotypes and traits over
three treatments are presented in Tables 1 and 2. The
highest mean values for all the traits were recorded

64 ~ M 19 Maydica electronic publication - 2019

 Drought screening in maize hybrids 4

Table 3 - Genotypic and phenotypic correlation among various observed characters and seed vigour indices (Control)

Characters EP SL RL SRL FW DW SV I SV II

EP G 1.000 -0.107 0.204 0.456* -0.131 -0.193 0.244 -0.021

P 1.000 -0.107 0.204 0.454* -0.131 -0.192 0.244 -0.021
SL G 1.000 0.508* 0.155 0.037 -0.143 0.790** -0.163
P 1.000 0.508* 0.154 0.037 -0.142 0.790** -0.161
RL G 1.000 0.487* 0.240 0.229 0.910** 0.263
P 1.000 0.485* 0.240 0.228 0.910** 0.262
SRL G 1.000 -0.077 -0.137 0.475* -0.055
P 1.000 -0.074 -0.131 0.473* -0.050
FW G 1.000 0.877** 0.146 0.879**
P 1.000 0.877** 0.146 0.879**
DW G 1.000 0.032 0.984**
P 1.000 0.033 0.984**
SV I G 1.000 0.073
P 1.000 0.073
SV II G 1.000
P 1.000

EP- Emergence percentage, PI- Promptness index, GSTI- Germination stress tolerance index, SL- Shoot length, SLSI- Shoot length stress index, RL-
Root length, RLSI- Root length stress index, SRL- Seminal root length, FW- Fresh weight, DW- Dry weight, SV I- Seed vigour I, SV II- Seed vigour II

*: Significant at 5% level; **: Significant at 1% level

in control and were reduced under PEG treatments.
Highest mean value for emergence percentage was
recorded by hybrid G5 (AUK-30) and G9 (AUMH-
8855) under PEG induced drought. Increase in PEG
concentration adversely affected seed germination.
Similar reduction in germination was reported by
Partheeban et al. (2017) and Khodarahmpour (2011) in
maize. Such reduction of EP is attributed to reduced
cell division and plant growth metabolism (Ayaz et al.,
2001).
Promptness index and germination stress tolerance
index are directly related to emergence percentage.
The hybrids G5 (AUK-30) and G9 (AUMH-8855)
showed higher mean values for both the characters.
The reduction in mean values due to PEG-6000
was also reported by Ahmad et al. (2015) in maize

Table 4 - Genotypic and phenotypic correlation among various observed characters and seed vigour indices (T2)

Characters EP SL RL SRL FW DW SV I SV II

EP G 1.000 0.981** 0.888** 0.927** 0.720** 0.751** 0.956** 0.988**

P 1.000 0.959** 0.869** 0.907** 0709** 0.743** 0.955** 0.987**
SL G 1.000 0.912** 0.937** 0.777** 0.788** 0.917** 0.959**
P 1.000 0.910** 0.935** 0.775** 0.785** 0.905** 0.935**
RL G 1.000 0.903** 0.900** 0.903** 0.818** 0.874**
P 1.000 0.902** 0.900** 0.901** 0.808** 0.854**
SRL G 1.000 0.714** 0.708** 0.881** 0.915**
P 1.000 0.714** 0.707** 0.870** 0.894**
FW G 1.000 0.972** 0.563** 0.672**
P 1.000 0.971** 0.560** 0.662**
DW G 1.000 0.607** 0.728**
P 1.000 0.605** 0.720**
SV I G 1.000 0.976**
P 1.000 0.974**
SV II G 1.000
P 1.000

*: Significant at 5% level; **: Significant at 1% level

EP- Emergence percentage, PI- Promptness index, GSTI- Germination stress tolerance index, SL- Shoot length, SLSI- Shoot length stress index, RL-
Root length, RLSI- Root length stress index, SRL- Seminal root length, FW- Fresh weight, DW- Dry weight, SV I- Seed vigour I, SV II- Seed vigour II

64 ~ M 19 Maydica electronic publication - 2019

 Drought screening in maize hybrids 5

Table 5 - Nei's original measures of genetic identity and genetic distance values
Pop.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
ID
1 **** 0.81 0.44 0.61 0.47 0.42 0.54 0.41 0.39 0.51 0.64 0.68 0.62 0.70 0.21 0.44 0.27 0.39 0.32 0.25 0.42
2 0.21 **** 0.41 0.57 0.60 0.31 0.46 0.43 0.49 0.60 0.64 0.60 0.62 0.62 0.32 0.41 0.21 0.29 0.36 0.18 0.31
3 0.83 0.90 **** 0.50 0.61 0.71 0.49 0.50 0.60 0.61 0.63 0.54 0.39 0.55 0.49 0.63 0.34 0.52 0.52 0.48 0.44
4 0.50 0.56 0.69 **** 0.65 0.40 0.58 0.48 0.50 0.73 0.65 0.48 0.51 0.51 0.53 0.42 0.30 0.46 0.44 0.44 0.32
5 0.76 0.51 0.49 0.43 **** 0.48 0.52 0.47 0.86 0.60 0.60 0.55 0.62 0.62 0.64 0.49 0.25 0.65 0.70 0.53 0.55
6 0.87 1.19 0.34 0.91 0.73 **** 0.57 0.42 0.67 0.48 0.65 0.70 0.50 0.67 0.31 0.53 0.46 0.49 0.51 0.56 0.47
7 0.63 0.78 0.71 0.55 0.65 0.56 **** 0.56 0.49 0.57 0.60 0.56 0.46 0.55 0.39 0.49 0.55 0.45 0.51 0.64 0.47
8 0.89 0.85 0.69 0.74 0.76 0.87 0.58 **** 0.48 0.60 0.39 0.34 0.57 0.49 0.30 0.26 0.31 0.28 0.27 0.50 0.50
9 0.94 0.71 0.50 0.69 0.15 0.40 0.71 0.74 **** 0.57 0.63 0.59 0.63 0.63 0.53 0.38 0.30 0.65 0.67 0.50 0.56
10 0.67 0.51 0.49 0.31 0.51 0.73 0.57 0.52 0.56 **** 0.72 0.51 0.54 0.46 0.40 0.49 0.29 0.33 0.49 0.53 0.39
11 0.45 0.45 0.46 0.43 0.52 0.43 0.50 0.95 0.46 0.32 **** 0.77 0.62 0.70 0.47 0.57 0.40 0.48 0.64 0.46 0.38
12 0.38 0.52 0.61 0.74 0.59 0.36 0.58 1.08 0.53 0.67 0.26 **** 0.74 0.74 0.32 0.59 0.47 0.54 0.57 0.46 0.42
13 0.49 0.48 0.93 0.67 0.48 0.69 0.77 0.55 0.46 0.62 0.49 0.30 **** 0.85 0.31 0.43 0.32 0.47 0.57 0.42 0.57
14 0.36 0.48 0.60 0.67 0.48 0.40 0.61 0.71 0.46 0.77 0.36 0.30 0.16 **** 0.38 0.59 0.40 0.59 0.62 0.49 0.68
15 1.55 1.14 0.71 0.63 0.45 1.19 0.93 1.21 0.63 0.92 0.76 1.14 1.18 0.95 **** 0.57 0.38 0.73 0.60 0.51 0.43
16 0.83 0.90 0.47 0.88 0.71 0.63 0.71 1.34 0.98 0.71 0.57 0.53 0.84 0.53 0.56 **** 0.64 0.67 0.74 0.60 0.52
17 1.32 1.57 1.08 1.21 1.39 0.79 0.61 1.17 1.21 1.23 0.92 0.76 1.14 0.91 0.98 0.45 **** 0.55 0.62 0.53 0.45
18 0.94 1.25 0.65 0.78 0.43 0.71 0.81 1.26 0.44 1.12 0.74 0.61 0.75 0.53 0.31 0.41 0.59 **** 0.78 0.52 0.72
19 1.15 1.01 0.65 0.83 0.35 0.67 0.67 1.30 0.39 0.71 0.45 0.57 0.55 0.49 0.52 0.30 0.47 0.24 **** 0.59 0.63
20 1.38 1.73 0.73 0.82 0.64 0.59 0.44 0.69 0.69 0.64 0.78 0.78 0.88 0.71 0.67 0.51 0.63 0.65 0.54 **** 0.69
21 0.87 1.16 0.82 1.14 0.60 0.75 0.75 0.69 0.58 0.94 0.98 0.87 0.57 0.39 0.84 0.65 0.80 0.33 0.47 0.37 ****

Nei's genetic identity (above diagonal) and genetic distance (below diagonal)

hybrids. Germination Stress Tolerance Index is directly
proportional to germination percentage and are
inversely related to water stress (Partheeban et al.,
2017).
Root length was found increased under water stress
conditions. A severe reduction in root length in PEG
concentration @ 20% was observed. The highest mean
values for root length were recorded by G5 (AUK-30),
followed by G9 (AUMH-8855). Drought resistance traits,
like extensive viable root system that could explore
deeper soil layers for water, are important criteria
for selection. A better root growth at seedling stage
would result in perfect root architecture at maturity
(Nejad, 2011). Shoot length and seminal root length
Fig. 1 - Distinct bands identifying drought tolerant hybrids
showed reduced mean values with the increasing PEG
concentration.
Drought stress tolerant maize genotypes
The stress tolerance indices such as shoot length
stress tolerance index, root length stress tolerance
index and vigour indices also recorded reduced mean
values under drought. The hybrid G5 (AUK-30) showed
highest mean values for all the stress tolerance indices.
Arisandy et al. (2017) studied PEG induced drought in
maize and reported that stress tolerance indices could
be used for selection of adaptive maize hybrids to
normal and drought stress conditions.
The mean comparison revealed that the hybrid
G5 (AUK-30) had higher significant values for nine
characters viz., emergence percentage, promptness
index, germination stress tolerance, shoot length
stress tolerance index, root length, root length stress
tolerance index, seminal root length, seed vigour
index I and seed vigour index II whereas the hybrid G9
(AUMH-8855) showed significant mean values for eight
characters. Hence these two hybrids are deemed as
drought tolerant. The hybrid G6 (A-3501) which showed
very low mean values for almost all the characters was
considered susceptible for drought.
Correlation between drought tolerance indices
Correlation studied revealed highly positive
significant correlation between root length and SV

64 ~ M 19 Maydica electronic publication - 2019

 Drought screening in maize hybrids
I, whereas for SV II except root length, every other
characters showed negative correlation in control. Dry
weight showed significant positive correlation with
SV II in control. The correlation coefficient estimates
under higher osmotic stress (T2) indicated that all the
six observed characters were positively correlated
to each other and to seed vigour indices of which
emergence percentage, shoot length, root length and
seminal root length were of higher magnitude (Tables
3 and 4). Indirect selection can be made on the basis of
characters viz., emergence percentage, shoot length,
seminal root length and root length at early growth
stage to screen a large population for drought stress.
These results are in agreement with the findings of
Khan et al. (2004) and Partheeban et al. (2017).
SSR marker analysis
The DNA samples were isolated from twenty one
maize hybrids using plant genomic DNA isolation kit
(Origin, Kerala). The hybrids were studied using fifteen
SSR markers and one SSR primer pair bnlg1346 was
rejected from analysis as it did not amplify. Remaining
fourteen primer pairs were amplified which generated
good and reproducible products for all the hybrids
(Fig, S5 - S8).
The banding pattern generated by drought linked SSR
markers showed no distinct bands separating tolerant
and susceptible hybrids except for the marker umc
1962. The primer umc1962 revealed two distinct bands
for hybrids G5 (AUK-30) and G9 (AUMH-8855) which
were identified as tolerant hybrids from PEG induced
screening (Figure 1). Similar studies were carried out by
Dubey et al. (2009), Bawa et al. (2015) and Maheswari
et al. (2016).
The genetic diversity indices for twenty one hybrids
are summarized in Table S4. The number of observed
alleles (Na) and effective number of alleles (Ne)
averaged across all loci ranged from 1.07 to 1.43. The
percentage of polymorphic loci ranged from 7.14%
to 48.86%. Based on Shannon’s information index (I),
higher diversity was observed in G8 (AUC-751). The
summary statistics of population genetic diversity
revealed that the gene or allelic diversity is low in the
maize hybrids under study. Hence the overall variation
is considerably less indicating the hybrids converging
origin.
The genetic identity and distance between samples
were recorded and are presented in Table 5. High
genetic distance was recorded between G2 and G20
(1.73) which again recorded the least value for genetic
identity. High genetic identity was exhibited between
G5 (AUK-30) and G9 (AUMH-8855) (0.86) which in turn
64 ~ M 19
6
recorded the least value for genetic distance. Nei’s
matrix indicated that G5 (AUK-30) and G9 (AUMH-8855)
were the most identical hybrids with eighty six percent
genetic similarities. Similar study was carried out by
Tanvir et al. (2018) in maize using ISSR markers.
The banding pattern of umc1962 and genetic identity
values support the conclusion derived from seedling
screening that G5 (AUK-30) and G9 (AUMH-8855) are
tolerant to drought conditions
Conclusions
From the above findings, it can be concluded that
the two maize hybrids G5 (AUK-30) and G9 (AUMH-
8855) are drought tolerant, whereas, G6 (A-3501) is
susceptible to drought. The study also revealed that
variation among genotypes for promptness index (PI)
and germination stress tolerance index (GSTI) was
found to be a reliable indicator of drought tolerance
in maize. Selection can be made on the basis of these
characters at seedling stage itself. The SSR primer
umc1962 identified tolerant hybrids with distinct bands
where the allelic diversity was low. Indications obtained
through seedling and marker screening for drought are
in agreement.
References
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