Abstract

Drought stress during the imbibition phase of germination is the primary reason for both
inhibition or delayed seed germination and seedling establishment. Rapid germination of seed
and emergence of seedling is pre-requisite for the good maize production under adverse
environmental stresses like salinity, drought soil and high and low temperature which has been
reported under seed priming in osmoticum such as mannitol, polyethylene glycol (PEG). To
enhance the pre sowing metabolic activities priming is done that ensures the high germination
percentage. An experiment will be conducted in Lab on new variety 940 of Maize primed with
Mannitol of 2%, 4%, 6% and 8% (w/v) concentrations with artificial drought condition created
by using different concentration of salt solutions viz. 1.5 bar, 4.9 bar, 10.3 bar, 17.6 bar to study
the effect of priming by mannitol on germination of maize in water stress environment
performed as factorial experiment under completely randomized design (CRD) with three
replications
Introduction
Background
Water stress due to drought is probably the most significant abiotic factor limiting plant and
also crop growth and development (Khalilli et al., 2013). Seed germination and early seedling
growth are potentially the most critical stages for water stress. Many studies on plant responses
to drought stress with regard to seed germination and seedling growth have been recently
reported, including studies of agricultural crops, Oryza sativa, Zea mays, Spinacia oleracea,
Amaranthus mangostanus and helianthus annuus (Ahmad et al., 2009). Uneven or poor
germination and subsequently uneven seedlings growth can lead to a great financial losses, by
reducing crop production and lower prices of uneven plant batches which can be addressed by
seed priming resulting increased speed and uniformity of germination.
Seed priming is a presowing strategy for influencing seedling development by modulating pre-
germination metabolic activity prior to emergence of the radicle and generally enhances
germination rate and plant performance. Seed priming is soaking of seeds in a solution of any
priming agent followed by drying of seeds that initiates germination related processes without
radical emergence (McDonald, 2000). There are reports that seed priming permits early DNA
replication, increase RNA and protein synthesis, enhances embryo growth, repairs deteriorated
seed parts and reduces leakage of metabolites. Seed priming is seen as a viable technology to
enhance rapid and uniform emergence, high vigor and better yields in some field crops (Basra
et al., 2002; Chiu et al., 2002; Harris et al., 1999; Murungu et al., 2004). Earlier works showed
that the success of seed priming is influenced by the complex interaction of factors including
plant species, water potentiality of the priming agent, duration of priming, temperature, seed
vigour and dehydration, and storage conditions of the primed seed. Germinating the seeds in
solutions of different water potentials is a convenient method to study the responses of seeds
at germination against low water potential (Naylor, 1992).

Objectives
Broad objective
 To evaluate the effect of seed priming by mannitol on germination characteristics of
maize in water stress environment.
 Specific Objectives
 To calculate the different germination parameters
 To find out the effect of seed priming by mannitol in water stress environment

1. MATERIALS AND METHODS

2.1. Experimental Site and Materials

The above research was conducted in the laboratory of Agronomy -Institute of Agriculture and
Animal Science, Lamjung campus. Seeds of maize (Zea mays) of variety NEW 940 were used.
2.2. Experimental Design and Procedure

The experiment was carried out in Two Factorial Complete Randomized Design (CRD) having
3 replications and 25 treatments.
Table 1: Treatment details
Factor A (Water Stress level ) Factor B (Seed Priming)
T1 Control (0 Mpa) M1 Control (0%)
T2 0.15 Mpa M2 2% Mannitol
T3 0.5 Mpa M3 4% Mannitol
T4 1.0 Mpa M4 6% Mannitol
T5 1.7 Mpa M5 8% Mannitol

Mannitol @ 2%, 4%, 6% and 8% (w/v) concentrations were used for seed priming. The
unprimed dry seeds were taken as control. Seed priming was done for 12 hrs. Following the
priming, seeds were dried back to their original moisture content at room temperature for 24
hours. Twenty five seeds each for each treatment were placed in Petri dishes. Water stress
condition was created by using different concentration of NaCl solutions viz. 0.15 Mpa,0.5
MPa, 1 MPa and 1.7 MPa and a treatment with tap water was taken as control. Seeds were
incubated in a germination chamber at 24°C for 6 days, with regular supervision.
2.3. Data Collection and Parameters

Seeds were considered germinated when there was a visible coleoptile protrusion of more than
5mm in length through the seed coat. The germinated seeds were counted at an interval of 12
hours for 6 days.
2.3.1. Germination Percentage (G %)

Percentage of germination was measured according to formula of Ahammad et al. (2014).

Germination percentage = no. of seeds germinated *100
Total no. of seeds used
2.3.2. Mean Germination Time (MGT)
Mean germination time was calculated using formula (Bewly et al., 2013)
MGT = Sum of Dn
Sum n

Where, Dn = the no. of days counted from beginning of germination

n= the no. of different seed lots
2.3.3 Germination energy (GE)
It is defined as the percentage of seeds germinated in 3 days i.e., 72 hours (Bam et.al.,
2006). Germination energy is the rapidity of germination and was measured by modifying the formula
of (Yan Li, 2008)
Germination energy = no. of total germinated seeds in NaCl concentration in 3 days
Total no. of seeds used for germination

2.3.4 Germination index (GI)

In the germination index, maximum weight is given to the seeds germinated on the first day
and less to those germinated later on. The least weight is given to the seeds germinated on the
last day. Therefore, GI emphasizes on both the percentage of germination and its speed. A
higher GI value denotes higher germination percentage and rate of germination (Benech
Arnold et al., 1991).
Germination index= (6*n1+ 5*n2 + 4*n3+ 3*n4+ 2*n5+ 1*n6)
Where, n1, n2, ……… n6 = no of seeds germinated on the first, second
and subsequent days until the last day of observation.

2.3.5. Relative Water Content (RWC)

Fresh weight of the seeds along with the shoot and root were measured. Then the petridishes
with the maize seedlings were filled with water and kept for 12 hours. After 12 hours the turgid
weight of the seeds along with the shoot and root were measured by removing the extra water
of the dish with the help of tissue paper. Then the seedlings were wrapped in the envelope and
kept in hot air oven. The dry weights of shoot and root were determined after drying at 72°C
for 24 h. Following formula was used (Weatherly, 1950)
RWC% = FW−DW ×100 where, FW, DW and TW are Fresh wt., Dry Wt. and Turgid wt.
TW−DW
2.3.6 Coefficient of velocity of germination
CVG indicates the rapidity of germination. It increases when the number of germinated seeds
increases and time required for germination decreases. Theoretically, the highest possible CVG
is 100. This would occur if all seeds germinated in the first day. It is calculated by the formula
given by (Jones and Sanders, 1987).
CVG = N1+ N2+ ……. NX/ 100 * N1T1+N2T2+ …… NXTX
Where, N= numbers of seed germinated each day,
T= No. of days from seedling corresponding to N

2.4. Statistical Analysis

The statistical analysis of data was done using computer software RStudio Version 1.1.463
applying 5% level of significance.