INTRODUCTION

The seeds of a large number of arid zone species fail to germinate because
of the impermeable seed coat and others because of their dormant embryos. The
morphology of seeds plays a vital role in germination (Sen and Chatterji, 1968).
Very little information is available on the comparative external morphological
characters, e.g. the shape, size and peculiarities of the surface and colourings of
the seeds of Indian desert. Probably in no other habitat a seed is subjected to such
rigorous environmental conditions as in the arid regions where an acute shortage
of water is a primary limitation in the germination of seeds.

Size and maturity of seed are important, and inseparable from viability and
vigor. The relative size and maturity of seeds relates directly to their survivability
when planted for highest viability and vigor, seeds must be large and fully mature.
Large, mature seeds have more stored food to nourish the seeds once they have
sprouted, and produce strong, vigorous seedlings, which have a better chance of
surviving and thriving under field conditions.

It is better to allow seeds to ripen to full maturity before they are harvested.
Mature seeds have higher germination and survival rates than seeds harvested at
an immature stage. This is because of natural metabolic changes the seeds
undergo as they mature, in preparation for dormancy. Small or mis-shapen seeds
are shorter-lived under storage conditions than larger, better-formed seeds. Small
seeds also contain less stored food to help them emerge from the soil and produce
healthy seedlings.

Although variations in seed shape are classically interpreted almost wholly

as adaptations, some features of shape may be thrust upon a seed by the
conditions inside the ovary in which it develops. Much of the great variety in seed
shape is related to dispersal. Seed colours are generally adaptive in respect to
visually orienting seed predators (Cook et al., 1971). In some species the colour of
seeds is known to maintain their viability for longer period, which may be probably
due to seed coat which acts as a filter under sunlight for the enclosed embryo,
keeping it exposed to far-red effects (Pandey, 1965).

Great variations in seed size are also found in the same genus and family.
In general, lighter seeds are found smaller in size, and heavier seeds are the
largest. As a seed on the surface of the soil exchanges water with the atmosphere,
and to absorb sufficient water to germinate, it must absorb water across the soil-
seed interface faster than it loses it across the seed-atmosphere interface. Large
seeds make poor seed-water contact and water uptake is slow as compared with
water loss. Large seeds may have difficulty in obtaining sufficient water because of
their low ratio of surface to volume. The ratio declines as seed size increases.

Germination and vigor are at their highest when seed is at its maximum dry
weight, a stage known as physiological maturity in most species (Tekrony and Egli,
1997); however, like any other form of life, they can not retain this identity
indefinitely. Seed is seldom planted immediately after harvesting, it is stored for
few days, weeks, months or years during which the seeds deteriorate, moving
inexorably towards death (Gregg et al., 1994). On account of biological activities
taking place in seed during storage, the seed deteriorates in quality resulting in
impairment of germination and vigour. The factors responsible for deterioration are
many. Seed viability in storage is determined not only by the period of storage but
also by the type of storage, storage environment and seed treatment if any
(Selvaraju and Krishnasamy, 2005). During storage viability is also affected by
various factors such as preharvest climatic conditions, temperature and relative
humidity of atmosphere (Harrington, 1963), seed moisture content (Vadivelu et al.,
1985), and packaging material (Katiyar and Dubey, 1992; Vadivelu et al., 1985).

During deterioration, vigor is the first component of seed quality, which is

lost, followed by a loss of germination capacity and viability (Trawatha et al., 1995).
Deterioration of seeds is an inexorable, continuous and irreversible process. Its
rate is largely determined by genetic factors, prestorage history of the seeds and
the storage environment in which they are kept. The rate of deterioration depends
upon various factors including both pre and post storage conditions. Since in many
cases there is a long gap between the time of planting and the time of seed
production, it is essential to store seeds under proper conditions to ensure high
germination and vigor upto its next planting season. The most widely accepted and
useful index of seed deterioration is reduction in viability. The germination
percentage of seed is maximum at the time of physiological maturity and later on
as the age increases, the germinability of seed declines. Viable seeds continue to
respire and metabolize during storage, leading to loss of viability.

The dry weather that prevails during seed maturation and harvest makes it
possible to harvest seeds, not only with low moisture but also with high initial
viability. There is a need to pay attention to the pre-storage, storage, and post-
storage phases to lower the risks of losses in viability during storage (Cabrera and
Lansakara, 1995).

The environmental conditions during seed maturation, threshing and the

location where the seeds are produced have a high impact not only on yield, but
also in seed moisture management and overall quality in terms of viability,
germinability, seed health, vigor and even plant performance. The critical factors in
storage are the kind of seed, seed moisture content, initial viability of the seed,
storage temperature, relative humidity of the storage area, duration of storage.

Seeds of some species are hygroscopic due to some chemical differences,

or due to physical differences that present resistance to moisture inflow to the
seeds (example hard seeds), or simply present a physical resistance to air flow
through the seed mass. Moisture content in the seed influences many factors, by
increasing metabolic activities, higher respiration, fungal problems, heating,
weakening and finally ending in the death of the seed. Seed moisture is not only
one more factor, but it is a causal factor of many other problems.
 For practical purposes, a moisture level below 13% is safe for storage of
most seeds. The lower the moisture content (below 13%), the longer seeds can be
stored, provided that the moisture level can be controlled throughout the length of
storage. Seed moisture content fluctuates with the changes in relative humidity,
which changes from the summer months (low RH) to the winter months (high RH),
(Justice and Bass, 1978). The magnitude of these fluctuations can vary with the
type of storage, type of bags used, and the kind of seeds, which influence the
migration of moisture from the air to the seed and vice-versa.

The high initial viability of seeds is one of the key factors that contribute to a
successful storage of high quality seeds. Seeds that have high initial viability
maintain their quality for longer period than seeds with low viability. Seeds are
hygroscopic, and can take up moisture from the air and release it back. If the
seeds are full, inside sealed containers, there is little air space and the moisture
content of the seed determines the relative humidity. If the seeds are in bulk
storage or in bags that allow air movement, seed moisture is determined by the
relative humidity of the air. The seeds adjust their moisture by trying to reach
equilibrium with the relative humidity. Temperature influences the amount of
moisture air can hold. It has been suggested that the sum of the percentage of
relative humidity plus the temperature in degrees Fahrenheit should not exceed
100 for safe storage (example: 50% RH and 50°F temperature; or 40% RH and
60°F; or 60%RH and 40°F), (McDonald and Copeland, 1999). The longer the
storage time needed, the lower these two factors should be.

Prolonged storage can lead to a gradual loss of vigor and finally a loss of
viability. The longevity varies among species, varieties, seed lots, and even among
individual seeds inside the same bag. Storage fungi have the capacity to grow at
very low seed moisture content and cause seed deterioration by producing toxic
substances that destroy the cells of seeds, which create dead tissue to sustain the
saprophytic fungi. Insects such as weevils can cause substantial damage to stored
seeds. The best prevention to these problems is by storing seeds at low moisture
during storage period.

Because of the complexity of all these factors during storage of seeds, this
research experiment is planned to study the morphological features of the seeds
and practices that needed to reduce the effect of storage under different conditions
on the viability of the seeds.

MATERIALS AND METHODS

Plant material:
The study on seed morphology, viability and storage was conducted on all of the
selected multipurpose species (MPS), viz. Acacia nilotica, Acacia tortilis, Albizzia
lebbek, Butea monosperma, Cassia angustifolia, Cassia siamea, Casuarina
equisetifolia, Cenchrus ciliaris, Plantago ovata, Prosopis cineraria, Prosopis
juliflora, Stylosanthus hamatha and Tecomella undulata.

Morphological features of seeds:

The data on seed colour, seed shape, texture of seed coat, length, width and
weight of seeds and imbibition of water by seeds were recorded. Seed colour and
shape were observed visually, seed coat texture was recorded by personal
discretion of touch, length and width were measured with the help of thread and
scale, weight was recorded electronically on digital balance.

Imbibition of water:
Imbibition of water by seeds was recorded by immersing 100 seeds of each
species in distilled water for 30 minutes and gain in weight of seeds after
immersion was recorded.

Determination of moisture content:

The moisture content of seeds was calculated by drying seeds in an oven for 24
hours at 1050C and expressed on fresh weight basis.

Drying of seeds:

Seeds were dried by exposing them for a couple of days in a warm, well ventilated,
but shaded place, before exposing them to sun for complete drying. The seeds
were spread in thin layers, 2-3 seeds thick, on wooden trays for drying. The seeds
were stirred or over turned periodically to expose all the surfaces of seed and
prevent excessive heating of any side.

Topographical Tetrazolium Test:

The method described by Moore (1962, 1973) and as prescribed in ISTA (1976),
was used to determine the viability of seeds through tetrazolium staining, for which
the seeds were soaked in distilled water for 24 hours. The seed coat was then
removed and seeds were soaked in 0.5% solution of tetrazolium chloride followed
by incubation for 24 hours at 37 0C in dark. Topographical patterns of formazan
stain were studied on fresh seeds as well as stored seeds after regular duration of
storage, under different conditions. Seeds having pink embryos were counted as
viable.

Storage of seeds:
After checking the initial viability of all the species followed by drying, all the seed
lots were divided into three parts. The seed lots were stored under ambient
conditions in:
 Air tight glass bottles
 Sealed polythene bags
 Muslin cloth bags
Seeds from all the storage containers were tested for their viability at bimonthly
interval upto 24 months.

RESULTS AND DISCUSSION

 Seeds of woody plants exhibit a great range of variation in shape, size,
colour and seed coat surface. These are very often adapted to the condition in
which the various species evolved (Khullar et al., 1991). Germination is a
complicated phenomenon in which the features of seeds like texture of seed coat,
seed size, seed weight, imbibition of water by seeds, etc. holds a great
significance. The morphological features of the seeds of each species viz. color,
shape, texture of seed coat, length, width and seed weight are given in Table 3.1.
The data reveals significant variations in morphological features among the seeds
of different species. Seed colour varied from creamish white in P. ovata to blakish
brown in A. nilotica. Shape of seeds varied from boat shape in P. ovata to flat in B.
monosperma. Seed coat texture varied from Smooth in P. juliflora to rough and
grooved in C. angustifolia.

The seed characters of different species studied for germination, varied.

Vanitha et al. (2004), have reported that seed size and colour grading have
significant influence on germination of seeds. It is also evident from several studies
that seed viability and germinabilty increases with the increase in seed size
(Murthykarvy, 1973; Manonmani and Vanangamudi, 2003). The potential for
diversity and modification of shape is greatly increased by the incorporation of
extra ovular structures in the dispersal units, as testa may be expanded to form
wings (Tecomella undulata) Seeds with hooked projections on the testa
(Stylosanthus hamatha), which permit dispersal by animals are rare, and where
wings or plumes are developed as testa outgrowths, the seeds are usually small.

The seed coat of Acacia tortilis was very hard and smooth, where as that of
Butea monosperma, Cenchrus ciliaris and Stylosanthus hamatha was soft (Table
3.1). Seed coats of other species were moderately hard. The length and width of
seeds also varied among different species. The seeds of B. monosperma were the
biggest with length and width rages of 34-44mm and 22-30mm respectively (Table
3.1), where as those of P. ovata were the smallest, with length ranges of 1.5-2 mm
and width ranges of 0.5-1 mm. All the other species lay within these minimums to
maximum ranges. Seed weight of C. equisetifolia was the least (0.0014-0.0017
gram per seed), where as that of B. monosperma was the maximum (0.961-
1.134g, Table 3.1).

Seed moisture content is the very important factor, which determines the
viability of the seed in storage. Determination of seed moisture content is very
essential in order to decide the extent to which the seed needs to be dried for
storage. In India, the tropical sun and air can provide excellent conditions for
preparing seeds for short-term storage. In such conditions the efficient storage of
orthodox seeds depends on the extent to which they can be dried. Drying should
not be too rapid, however because, if moist seeds are subjected to high
temperatures, their viability may be seriously effected (Khullar et al., 1992).

The most important, urgent, and crucial requirement when the seed is
harvested is to measure the moisture content to see if it is at a safe level. When
seeds have high moisture it has to be dried. The data on the imbibition of water by
seeds in 30minutes and moisture content of seeds of selected species before and
after drying are presented in Table 3.2. Seeds stored with higher moisture content
exhibit increased respiration, which leads to fungal invasion and poor seed viability
and vigour.

It is evident from Table 3.2, that the moisture content of Butea monosperma,
Cassia angustifolia and Plantago ovata was very high (14.6, 14.02 and 15%
respectively), while, that of Albizzia lebbek and Stylosanthus hamatha was low
(8.23 and 8.68% respectively). After air-drying the moisture content of the seeds
reduced significantly in all the species. The reduction was more than 15 percent in
B. monosperma, C. angustifolia, C. siamea, P. cineraria, P. juliflora and T.
undulata. Where as, in A. tortilis, P. ovata and S. hamatha, the percent moisture
reduction is less than 8%. The imbibition of water per seed was minimum for T.
undulata (0.009g) and maximum for B. monosperma (10.28g). Seeds of rest of the
species gave imbibition within these minimum to maximum ranges.

Table 3.2: Imbibition of water by seeds and moisture content of seeds of MPS
before and after drying.

Species Imbibition of Moisture Moisture Percent

water per content content after reduction
seed (g) before drying drying (%) in seed
(%) moisture
content
A. nilotica - 11.05 09.88 10.58

A. tortilis 0.0230.0021 10.21 09.37 08.23

A. lebbek 0.2190.0151 08.23 07.35 10.69

B. monosperma 10.2760.143 14.60 13.01 17.74

C. angustifolia 0.0630.0079 14.02 11.06 17.26

C. siamea 0.0480.0022 12.84 10.22 20.40

C. equisetifolia - 13.94 11.85 14.99

C. ciliaris 0.00350.007 10.73 09.18 14.44

3
P. ovata - 15.00 13.91 07.26

P. cineraria 0.0530.0026 11.34 09.63 15.08

P. juliflora - 12.78 10.84 15.18

S. hamatha 0.0640.0029 08.68 07.99 07.95

T. undulata 0.0090.0015 11.63 09.41 19.09

Some seeds are viable for very short period under ambient conditions. Seed
viability and storability are predominantly dependent on seed moisture and storage
temperatures (King et al., 1981). High seed moisture and high storage temperature
promote the metabolism and microbial growth causing loss of viability (Dijode,
2003). Reduction in seed viability is associated with loss of vigour and changes at
cellular level causing excessive leaching of metabolites (Givelberg et al., 1984).
Seed storage may influence seed viability and reduces seed vigour. The extent of
the change depends on the time span and storage conditions i.e., temperature and
humidity (Dell”Aquilla, 1987). Type of container used for storage also influences
the seed viability.

Vanangamudi (2001) has emphasized that quick viability testing is very

important in forest seeds, which require longer time to germinate or may exhibit
varying types of dormancy or for seeds which lose their viability very quickly. For
rapidly obtaining an indication of germination potential and viability of seeds,
topographical tetrazolium test is very useful (Agrawal, 1985). When seeds are
incubated for 4-16 hours in tetrazolium chloride solution, dehydrogense enzyme, in
actively respiring areas of the embryo and endosperm, produce hydrogen ions,
which react with the colorless tetrazolium chloride to form the red colored non-
diffusable formazan. By evaluating the location and intensity of the formazon
stains, healthy and damaged parts of the seed can be differentiated (Khullar et al.,
1991). The data on initial viability of the seeds of all the selected species and their
viability at bimonthly interval upto 24 months of storage under different storage
conditions is presented in Table 3.3. The data reveals highest initial viability
(100%) in the seeds of A. lebbek and B. monosperma, and lowest initial viability in
the seeds of P. ovata. Initial viability of rest of the species lies between these
maximum to minimum ranges. The data on initial viability of seeds of all the
selected multi purpose species are represented in Figure 3a. Viability under
different storage conditions after 6 months of duration is presented in Figure 3b. It
reveals that seeds of all the multipurpose species, which were stored in muslin
cloth bags, exhibits maximum decline in seed viability. The data on viability of
seeds after 24 month of storage duration are presented in Figure 3c. After the
storage of 24 months, there was 100 percent decline in the viability in the seeds of
B. monosperma, C. equisetifolia, C. ciliaris, P. ovata, S. hamatha and T. undulata
under all storage conditions.
 Data indicates the declining pattern of viability with storage period in
different species under different storage conditions. The viability of seeds is
maximum at the time of physiological maturity and later on as the age increases,
the viability of seeds declines because viable seeds continue to respire and
metabolize during storage, leading to loss of viability (Tewari, 1976).

Seeds of A. nilotica, which initially had 91% viability (Table 3.3), retained
more than 79% viability after 24 months of storage in glass bottles and polythene
bags, while the seeds stored in muslin cloth bags showed 60% viability after 24
months of storage (Figure.3c and 3d). The seeds kept in glass bottle and polythene
bags retained the viability 1.3 times more than the seeds kept in muslin cloth bags.
Seeds stored in glass bottle showed nearly similar percentage viability as observed
in seeds stored in polythene bags. Puri and Gargya (1995) have reported that
seeds of A. nilotica are viable upto 12 months. Prakash et al. (2003), have
suggested that properly collected, dried, clean seeds of A. nilotica can be stored in
gunny bags, tin or baskets in a cool dry place. They reported that, if these seeds
are stored in airtight containers, there is a little loss in viability upto 3 years.
Vanangamudi (2001) has emphasized that for A.nilotica species, acid scarification
for 60min. followed by 12 hour water soaking and longitudinal splitting of
cotyledons is the best preconditioning method for viability assessment.

In case of A. tortilis initial viability was 78%(Table 3.3), which progressively

declined during storage. After 24 months of storage in glass bottle and polythene
bag, 67% and 65% viability was recorded respectively, while only 50% (Figure.3c
and 3e) viability was observed in seeds stored in muslin cloth bag. As mentioned in
the Information Brochure on A.tortilis species by ICFRE, Dehradun, A.tortilis seeds
retain their viability for 7-8 years and present few storage problems by virtue of
their hard seed coat which, restricts moisture exchange and loss of store reserves
through respiration. But contrary to this, Puri and Gargya (1995) have reported that
A.tortilis seeds are viable for 12-24 months. Seeds should be stored in dry and
moisture proof airtight containers like glass or plastic bottles. In the present study,
storage of seeds in glass bottles and polythene bags retained viability for longer
period than in muslin cloth bag.

Seeds of A. lebbek showed 90% viability after 24 months of storage in glass

bottles and 70% viability in polythene bags (Table 3.3; Figure 3f), while only 50%
viability was observed in seeds stored in muslin cloth bag for the same period.
Storage of seeds in glass bottle and polythene bag retained viability for longer
period than in cloth bag. Nipping off the seedcoat followed by 12hour soaking and
longitudinal splitting of cotyledons to be the best suited preconditioning method for
A. lebbek (Vanangamudi, 2001). Prakash (1998), reported that seeds of A. lebbek
can be stored for 4 to 5 years in sacks or polythene bags. Similar findings were
reported by Puri and Gargya (1995), who have reported that these seeds are
viable for 48-60 months.

The seeds of Butea monosperma had 100% viability (Table 3.3, Figure 3a
and 3g) when checked immediately after procurement, but the viability declined
sharply during the storage. The viability remained only 30%, 22% and 17% in glass
bottle, polythene bag and muslin cloth bag respectively after 12 months of storage
(Table 3.3) and not even a single seed remained viable upto 24 months of storage.
However, Prakash (1998), reported 90% viability in the fresh seed lots of Butea
monosperma and found that their pods can be stored for one year in sealed tins.

Initial viability of C. angustifolia seeds was 92%, which declined to 69%,

59% and 38% after 24 months of storage (Table 3.3, Figure 3c and 3h) in glass
bottle, polythene bag and muslin cloth bag respectively. In this species, although
viability declined with the storage period, the reduction was least in seeds packed
in glass bottle and it was maximum in seeds packed in muslin cloth bags.

In C. siamea, 84%, 80% and 41% viability was recorded after 24 months of
storage (Table 3.3, Figure 3c and 3i) in glass bottle, polythene bag and muslin
cloth bag respectively, while initial viability was 98% (Table 3.3, Figure 3a). The
viability reduction was 14%, 18% and 58% in all the respective containers. Prakash
(1998), reported that germination capacity of C. siamea seeds is 80% and these
can be stored for about 3 years in gunny bags or in sealed tins. Puri and Gargya
(1995) have also estimated their viability period to be 24-36 months.

Even though Casuarina equisetifolia is a very important multi purpose

species for arid and semi arid areas, its seeds are very sensitive and showed only
38% initial viability which declined rapidly and after 6 months of storage only 7%
and 3% (Figure 3b and 3j) viability was recorded in seeds stored in glass bottle and
polythene bag respectively. The findings of this study are supported by the findings
of Prakash (1998), that the seeds of Casuarina equisetifolia can not be stored for
long. Thus the seeds of this species must be sown as early as possible after seed
collection.

The viability of Cenchrus ciliaris was 48% at the start of study. But after only
6 months of storage the viability declined to 2% and 1% (Figure 3b and 3k), in
glass bottle and polythene bags respectively. Thereafter no seed remained viable
during further storage.

In Plantago ovata initial viability was only 40% (Table 3.3; Figure 3a and 3l),
which declined very rapidly in the seeds stored in muslin cloth bags. In glass
bottles only 9% seeds were found to be viable after 24 months of storage. Where
as in polythene bags 100% decline in viability was recorded after 10 months of
storage.
.
Initial viability of P. cineraria was 99%(Table 3.3, Figure 3a and 3m), which
was declined to 85% and 71% after 24 months of storage (Table 3.3, Figure 3c) in
glass bottle and polythene bag respectively, whereas, in muslin cloth bag it was
only 47% after 24 months of storage. Puri and Gargya (1995), found that P.
cineraria seeds are viable for 24 months and Prakash (1998), emphasized that dry
and clean seeds of P. cineraria can be stored in airtight containers in a dry place
for several years.

Seeds of P. juliflora, which initially had 85% viability (Table 3.3, Figure 3a
and 3n), retained 65% and 53% viability after 24 months of storage (Figure 3c) in
glass bottles and polythene bags respectively, while the seeds stored in muslin
cloth bags showed 37% viability after 24 months of storage. Prakash (1998) have
reported that viability of fresh seeds of P. juliflora is 80-90% and well dried seeds of
P. juliflora can be stored for about 2 years. Puri and Gargya (1995) have also
reported its viability period to be 18 months.

Stylosanthus hamatha is considered to be a boon to arid areas with initial

viability of 50% for freshly collected seeds (Table 3.3, Figure 3a and 3o). The
seeds are viable for very small time span. When stored in airtight containers like
glass bottle only 12% seeds were found to be viable after 8 months of storage.

Initial viability of Tecomella undulata seeds was 80% (Table 3.3, Figure 3a
and 3p), which declined very sharply. After 6 months of storage in glass bottle only
12% (fig 3b) seeds were found to be viable. Jindal and Pancholy (1994), have
worked on the effect of different seed containers on germination and seedling
characters in T. undulata. They reported that seed germination ranged from 27.1%
when seeds are stored in paper bags to 64.3% when stored in polythene bags.
Better germination was recorded in polythene bags and glass bottles than when
the seeds were stored in cloth bags, in the conventional way of seed storage.

The type of container had significantly influenced seed viability. The

superiority of glass bottles over polythene bags and again the superiority of
polythene bags over muslin cloth bags were evident at all intervals of storage.
Under ambient storage conditions in cloth bags a drastic reduction in viability was
observed in all the species after storage of 24 months. Seeds stored in cloth bags
lost their viability earlier than those stored in polythene bags and glass bottles due
to absorption of moisture. In case of airtight containers, i.e., polythene bags and
tin, there is no free exchange of gases, whereas in others, such as cloth bags or
paper bags, there is free exchange of gases (Jindal and Pancholy, 1994).

In glass bottles and polythene bags, the rate of metabolic activities during
storage may be reduced due to availability of limited air. It may be helpful in
delaying the aging process and ultimately maintenance of good viability,
germination and seed vigour in seeds stored in air limited conditions.