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Journal of Agricultural Science; Vol. 8, No.

10; 2016
ISSN 1916-9752 E-ISSN 1916-9760
Published by Canadian Center of Science and Education

Influence of Seed Storage Techniques on Germinability and Storability


of Cowpea (Vigna unguiculata (L) Walp)
Hillary Mireku Bortey1, Alimatu Osuman Sadia1 & James Yaw Asibuo1
1
CSIR-Crops Research Institute, Fumesua, Kumasi, Ghana
Correspondence: Alimatu Osuman Sadia, CSIR-Crops Research Institute, P. O. Box 3785, Fumesua, Kumasi,
Ghana. Tel: 233-026-484-1081. E-mail: [email protected]

Received: June 3, 2016 Accepted: August 2, 2016 Online Published: September 15, 2016
doi:10.5539/jas.v8n10p241 URL: http://dx.doi.org/10.5539/jas.v8n10p241

Abstract
This study was set to investigate the germinability and storability of the newly released cowpea genotypes and
secondly the influence of different storage materials under ambient storage condition on seed vigour and
germination over time. The experiment was laid in a Split-split plot design with four replicates. The main plot was
duration (0, 30, 60 and 90 days), while Storage material (polyethylene bag, cotton bag and glass container) was sub
plot and the four cowpea genotypes were sub-sub plot. Results from the study showed that irrespective of the
cowpea genotypes and storage material used, percentage vigour and germination were significantly affected with
time in storage. Seeds stored in cotton bags had the least percentage vigour (57%) and germination (65%) while the
highest was recorded for seeds stored in air-tight glass containers. Among all the cowpea genotypes studied,
Hewale was found to have poor storage abilities. For better storage of cowpea seeds for a period exceeding 3
months, it is preferable to use glass containers or black polyethylene bags even under ambient condition to
maintain seed vigour and germinability.
Keywords: cowpea, seed longevity, seeds storage, storage material
1. Introduction
Although seed quality is governed by their genetic make-up, the quality of seeds however, is deteriorated during
storage (Barua et al., 2009; Al-Yahya, 2001). Thus, seed storage and retention of seed viability has always been an
important consideration in agricultural production. Most seeds have been reported to be hygroscopic in nature, and
tendency of absorbing environmental moisture during storage is high (Copeland & McDonald, 1995). Poor storage
condition gives rise to deterioration of seed quality and the resultant loss of viability.
During storage, seed quality can remain at the initial level or decline to a level that may make the seed
unacceptable for planting purpose. Several environmental factors have been reported to affect seed viability
during storage (Rindels, 1995). Some of the factors that affect the longevity of seeds in storage could be the
genotype of seed, initial seed quality, storage conditions, and moisture content among others. Within the same
plant species, different varieties may exhibit different storing abilities either from genetic variations or other
external factors (Simic et al., 2007). However, irrespective of the initial seed quality, unfavourable storage
conditions, particularly air temperature and relative humidity contribute to accelerating seed deterioration
(Heatherly & Elmore, 2004). High relative humidity and temperature cause high moisture content in seeds and
result in low germination at the end of storage (McCormack, 2004).
In Ghana, most smallholder farmers store their seeds in various containers/materials including: pieces of cloth,
black polyethylene bag, clay pots, plastic containers, mostly under ambient conditions (Bortey et al., 2011). This
results in loss of seed viability over a short period of time depending on the type of crop seed (Barua et al., 2009;
Bortey et al., 2011). Additionally, several studies have indicated the effect of storage containers/materials on the
quality of seeds in terms of germination and viability over a period of time. A study conducted by Bortey et al.
(2011) indicated that tomato seeds stored in piece of cloth and tin containers resulted in low percentage
germination and vigour compared to seeds stored in glass containers. However, it has been reported that the
intensity of quality decreasing of stored seed under different storage techniques differ among plant species and
within plant species (Al-Yahya, 2001; Guberac et al., 2003). Thus, the choice of material/container used in storing
seed is crucial in ensuring that the quality of seed is maintained during storage. Moreover, since the effect of these
storage materials on the quality of seeds of different crops may vary, it is important to investigate and establish

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the most suitable storage material and condition for various food crop seeds. This would provide seed producers,
breeders and farmers information on how to maintain the integrity of the seed during storage.
This study was set to investigate the storability of the newly released cowpea genotypes and secondly the influence
of storage techniques on seed germinability and storability. Specifically, to investigate the influence of different
storage materials under ambient storage condition on seed vigour and germination of the newly released cowpea
genotypes.
2. Materials and Methods
The investigation was carried out at the Seed Technology Unit Laboratory of CSIR-Crops Research Institute
(CSIR-CRI), Kwadaso. Seed samples were collected from the Legumes Division of CSIR-CRI, Fumesua
approximately 3 weeks after harvesting. Four genotypes of cowpea seed samples used for the study comprised
Hewale, Vidiza, Asomdwoe (newly released) and Asenetapa (old variety), all unpigmented seed coat. Seed samples
were initially stored in a cold room for after harvesting before it was sampled for the experiment. Seed samples
were stored in polythene bag (black), Cotton bag (cloth), and Air-tight glass-bottle (Kilner jar) and kept under
ambient storage conditions for storage.
2.1 Monitoring of Storage Condition
Data on relative humidity and temperature was recorded for the storage period under ambient condition using an
analogue SeedBuro Hygrometer. Both temperature (degree Celsius) and percentage relative humidity were
recorded three times during the day for the entire study period. The average, minimum and maximum temperatures
and relative humidity at the end of the storage period were computed. Before seeds were kept in the various
containers, initial seed quality characteristics including percentage seed moisture (SMC), 1000 seed weight, seed
germination percentage (SGP), percentage seed vigour and dead seeds were recorded. Seed samples were taken
every 30 days from the various containers and the above stated seed quality characteristics recorded for 90 days.
2.2 Determination of Seed Germination Percentage (SGP)
Seed germination was tested on four 50 seed replicates for all treatments. Each replicate was set to germinate at
25± 2 °C on top of Non-toxic Paper towel, moistened and placed in a plastic seed box (Figure 1). Germination,
judged by the appearance of the radicle, was counted daily up to 8 days. At the final count, the number of normal
and dead seedlings was assessed. The SGP was calculated as follows:

number of germinated seeds


SGP (%) = × 100 (1)
Total seed number

90

80
% Seed Vigour/ Germination

70

60

50

40

30
0 day 30 day 60 day 90 day
Duration of Storage
% Seed Germination % Seed Vigour

Figure 1. Effect of storage duration on percentage seed vigour and germination

2.3 Determination of Percentage Seed Vigour


The Percentage Seed vigour was calculated by dividing the number of seeds that germinated at first count after 5
days by the total number of seeds sown.

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2.4 Seed Moisture Content (SMC)


SMC determined by the high constant temperature oven method at 130 °C for one hour according to
International Seed Testing Association (ISTA, 2007) procedures. 15 g of seeds was milled to a coarse texture
after which two replicates of 5 g of the coarsely milled cowpea was weighed and put in aluminum analytical
container (9 mm in diameter). After the oven reached 130 °C, the seeds in the container were put in the oven for
drying. After 1 hour drying, seeds were immediately removed and kept in a desiccator containing silica gel for
cooling. Seeds were re-weighed and the weight difference was used to compute the seed moisture content using
the formula: SMC (%) = (M2 – M1)/M2 × 100 where M1 is seed sample weight after drying, M2 is seed weight
before drying. Maximum tolerance was 0.2 between replicate.
2.5 1000 Seed Weight
For 1000 seed weight determination, eight replicates of 100 seeds were counted and weighed on a sensitive
balance and average weight was multiplied by 10.
2.6 Experimental Design and Analysis
The experiment was laid in a Split-split plot design with four replicates. The main plot was duration (0, 30, 60 and
90 days), while Storage material (polyethylene bag, cotton bag and glass container) was sub plot and the four
cowpea genotypes were sub-sub plot. A total of 192 samples consisting of 48 treatment combinations replicated
four times were prepared. Analysis of variance (ANOVA) and least significant difference (LSD) at 5% were
utilized to analyze the data using SAS (2007 ed.).
3. Results
The storage condition was relatively warm with an average temperature and humidity of 32 °C and 81%
respectively.

Table 1. Storage condition, Initial Seed Moisture Content and 1000 seed weight (g)
Storage condition Minimum Maximum Average
Temperature, ºC 28 34 32
Relatively Humidity (%) 74 88 81

Seed of cowpea genotypes used for the study had initial seed moisture contents ranging from 9% to 10.7%. While
Asetenapa recorded the least per cent seed moisture, Asomdwoe had the highest per cent seed moisture (10.7%).
Asomdwoe was observed to be small seeded compared to the other genotypes. It recorded the least 1000 seed
weight of 136 g while Videza recorded 169 g.

Table 2. Initial Seed Moisture Content and 1000 seed weight (g)
Cowpea Genotypes Initial Moisture Content 1000 Seed weight (g)
Hewale 9.8 149.4
Videza 10.4 169.5
Asomdwoe 10.7 136.3
Asetenapa 9.0 162.5

3.1 Effect of Storage Duration on Percentage Seed Vigour and Germination


Highly significant difference (P < 0.001) was observed among the various storage time on percentage seed
vigour and germination. Germination and seed vigour percentage was found to decrease with the increase in
storage duration (Figure 1). The mean percentage seed vigour and germination irrespective of storage containers
(polythene bags, cotton bags and glass containers) and genotype was found to decrease continuously from initial
78 to 34 % and 79.8 to 58.8 % after 9 months storage respectively (Figure 1).
3.2 Effect of Storage Material and Duration on Seed Percentage Germination
Storage material significantly (P < 0.001) affected the percentage of seed vigour and germination. Highly
significant (P < 0.001) difference was observed in mean vigour and germination percentages among cowpea
genotypes stored in different storage materials. The seeds stored in glass containers which had the highest vigour

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(63.7%) also had the highest germination percentage (72.5%), followed by seeds stored in polyethylene bags
(black) which had vigour percentage of 63% and mean germination percentage of 71.7% (Figure 2). The lowest
percentage seed vigour and germination was recorded by seeds stored in cotton bags (57%) and 65% respectively.
The results also indicate that there was a strong positive correlation (r = 1.00) between percentage seed vigour
and germination percentage (Table 3).

80.0

70.0
% Seed Vigour/Germination

60.0

50.0

40.0

30.0

20.0

10.0

0.0
Polybag Cottonbag Glass Container
Storage Materials used in storing cowpea seeds

% Seed Germination % Seed Vigour

Figure 2. Effect of Storage materials on seed percentage vigour and germination

Table 3. Correlation coefficients (r) of physiological and physical traits of cowpea genotypes after 90 days
storage
%Germ %Vigour SMC Seed_Wt
% Germ
%Vigour 1.0000
SMC 0.3877 0.3877
Seed Wt 0.2428 0.2428 -0.4157

3.3 Performance of Cowpea Varieties in Storage of Time


Varietal differences during storage in the present study were highly significant (P < 0.001). Among the newly
released varieties, Hewale stored poorly, irrespective of the packaging material and storage condition. Hewale,
decrease from 75% germination initially to 67% below acceptable minimum germination within 30 days of
storage and further decreased sharply at 60 days of storage to 47%. On the other hand, Videza and Asomdwoe
performed better in storage compared to an old cowpea variety, Asetenapa up to 60 days in storage. It was
however, observed that all varieties fell below minimum percentage seed germination for cowpea at day 90.
3.4 The Combined Effect of Storage Material, Crop Genotype and Duration on Percentage Seed Germination
It can be deduced from Figure 3, that combined effect of storage material cowpea genotype and storage time
were highly significant (P < 0.001). All cowpea genotypes irrespective of the storage materials declined in
germination percentage over time. Consistently, cotton bags used in storage also performed poorly among the
storage materials. Similarly, the genotype Hewale, irrespective of the storage material and duration of storage
decreased in percentage germination. The lowest percentage germination of 22% was recorded at 90 days in
cotton bag.

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Day 0 Day 30
90 90
80 80
70 70

Germination %
Germination %

60 60
50 50
40 40
30 30
20 20
10 10
0 0
Asetenapa Videza Hewale Asomdwoe Asetenapa Videza Hewale Asomdwoe
Cowpea Genotypes Cowpea Genotypes

Poly bag Cotton bag Glass container Poly bag Cotton bag Glass container

Day 60 Day 90
90 90
80 80
70 70
Germination %

Germination %
60 60
50 50
40 40
30 30
20 20
10 10
0 0
Asetenapa Videza Hewale Asomdwoe Asetenapa Videza Hewale Asomdwoe
Cowpea Genotypes Cowpea Genotypes
Poly bag Cotton bag Glass container Poly bag Cotton bag Glass container

Figure 3. Combined effect of storage material, genotype and duration on percentage germination

4. Discussion
4.1 Effect of Storage Duration on Percentage Seed Vigour and Germination
Across the cowpea genotypes studied, percentage seed vigour and germination reduced over time during storage
(Figure 1). This observation could be attributed to the relatively high storage conditions (temperature and
relative humidity) recorded during the period of storage as indicated in Table 1. The combined effect of
temperature and seed moisture content and relative humidity of storage conditions are main known factors
influencing seed deterioration and viability loss. These factors are applicable irrespective of the seed type as
observed in the present study. Eluid et al. (2010) in studying the effects of temperature and relative humidity on
the viability of bean seeds stored under stockists store condition observed that seeds stored under mean maximum
temperature and relative humidity of 30.8 °C and 80.1% respectively showed a rapid decrease in viability of bean
seed (Phaseolus vulgaris) and went below the accepted levels after one month of storage.
Further, it is generally known that seed deterioration is inevitable but the rate can be reduced when the key
storage factors such as temperature and humidity during storage are well managed (Ellis & Roberts, 1981;
Rindels, 1995; Adetumbi, 2009). With the exception of Hewale genotype, which maintained good germination up
to 30 days, the cowpea genotypes in the present study were able to store up to 60 days still with relatively good
germinability but was short below acceptable levels according to Ghana Seed regulation standards. Thus, as
observed by other authors (Pradhan & Badola, 2008; Adetumbi et al., 2011; Kamara et al., 2014), it can be
re-emphasized that irrespective of the initial seed quality, unfavourable storage conditions, particularly air
temperature and relative humidity contribute to accelerating seed deterioration (McCormack, 2004; Heatherly &
Elmore, 2004).
4.2 Effect of Storage Material on Percentage Seed Germination
Seed stored in cotton bag under ambient temperature and humidity recorded the lowest vigour and germination
percentage after 60 and 90 day period irrespective of variety. However, seeds stored in polyethylene bag and
air-tight glass container recorded high seed vigour and germination percentage. This present studies are in line
with reports of Asiedu and Powel (1998), Bortey et al. (2011) and Kamara et al. (2014). The author reported that

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seeds stored in humid and warm environments tend to absorb moisture from the surroundings, leading to
increased seed moisture content until equilibrium is established. Gurmit and Hari (1992) also made similar
observation and indicated that seeds stored in vapour proof containers maintain the desired quality of seeds for
longer periods than moisture pervious containers like cotton bags. When there is minimal gas exchange between
the storage material and surrounding environment, biochemical activities reduces and that could reduce the rate
of deterioration in seeds.
4.3 Cowpea Genotype Performance in Storage Over Time
Seed deterioration is a serious problem in developing countries where seeds are stored in places usually without
a proper control of humidity and temperature. Generally, all cowpea seed genotypes fell below minimum seed
germination percentage of 70% according to the Seeds (Certification and Standards) Regulations of Ghana after
90 days of storage (Figure 4). The various released cowpea genotypes stored significantly different. Among the
newly released genotypes, Asomdwoe and Videza stored well up to day 60 but comparable to the old Asetenapa
genotype while Hewale stored poorly in terms of percentage seed germination at day 60. Differences in the
storage potential of cultivars within a species have also been reported in grain legumes as being associated with
seed-coat pigmentation. To corroborate with this assertion, the findings of Asiedu and Powell (1998) in Cowpea
(Vigna unguiculata L. (Walp)), Abdullah et al. (1993) in Phaseolus vulgaris and Mugnisjah et al. (1987) in
soybean, reported that soybean cultivars having unpigmented seed coats deteriorated more rapidly during storage
than cultivars with pigmented seed coats. In the present study however, all cowpea varieties had light coloured or
unpigmented seed coat. This may rule out the influence of seed coat colour or pigmentation on the longevity of
the cowpea genotypes studied. This notwithstanding, the rate of seed ageing depends on the physiological status
and genetic constitution of the seeds as well as on the capacity of a cultivar to withstand ageing conditions
(Priestly, 1986; Kalpana & Madhava Rao, 1995). Progressive loss in seed quality attributes occurred with ageing
in all varieties but at a faster rate with the Hewale genotype. The observed reduction in percentage seed
germination over time could also be linked to the reduction in enzyme activity within the seed Iqbal et al. (2002),
Ruzrokh et al. (2003), and Demirkaya et al. (2010).

90

80
% Seed Germination

70

60

50

40

30
0 day 30 days 60 days 90 days
Duration of Storage
Asetenapa Videza Hewale Asomdwe

Figure 4. Performance of Cowpea genotypes in storage over time

5. Conclusion
Results from the present study suggest that cowpea genotype differences affect germinability and storability; that
each genotype behaved differently during storage. It further indicates that glass containers and polyethylene bags
are suitable seed storage materials for cowpea compared to cotton bags. The storage material significantly
affected the quality of seed in terms of percentage vigour and germination.
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