J. Env. Bio-Sci., 2015: Vol.

29 (2):429-435
(429) ISSN 0973-6913 (Print), ISSN 0976-3384 (On Line)

IMPACT OF SEED WEIGHT AND PRE-SOWING TREATMENTS ON GERMINATION AND

SEEDLING GROWTH OF CALOPHYLLUM INOPHYLLUM L.
Vikas Kumar1, Ajeesh, R. and Jijeesh, C.M.
1
Department of Silviculture and Agroforestry, College of Forestry, Kerala Agricultural University, KAU, Thrissur, Kerala (India)
[Corresponding author E-mail1: vkskumar49@gmail.com]

Received: 18-06-2015 Accepted: 21-10-2015

In the era of climate change, there is an increasing attention on alternate fuels especially towards biodiesel production.
Calophyllum inophyllum L. has been identified as the most suitable feedstock for future generation biodiesel. However, its large
scale cultivation is often constrained by the availability of good quality planting stock. The present study evaluated the response
of size graded C. inophyllum seeds to different pre-sowing treatment and to evaluate the initial nursery performance of seedlings
in the size graded seeds. The seeds were graded into size categories 2-5 (small), 5-8 (medium) and 8-11 g (large) based on their
weight. The seeds belonging to different weight classes were subjected five pre-sowing treatments. It indicated that the
germination parameters of the size graded seeds significantly influenced by the pre-sowing treatments and the seeds belonging
to medium weight class without hard seed coat recorded the highest germination (83.3 %) which was followed by the large
weight class. Size grading significantly influenced the seedling growth and biomass production and the performance of the
seedlings was in the order large> medium> small. The various physiological, growth analysis and quality indices also supported
the higher growth and biomass production of the large seedlings.

Many trees, shrubs and herbs available plentifully in India MATERIAL AND METHODS
can be successfully exploited for biodiesel production. A large
The present study was conducted at the College of Forestry,
number of native species have been assessed for growth on
Kerala Agricultural University, Thrissur, Kerala, India during
degraded land in India which produces biodiesel at appreciable
July 2013 to June 2014. The nursery area is located 40 meters
quantities. Among them, Calophyllum inophyllum L. has been
above MSL at 10032’N latitude and 76026’E longitude. The
identified as the most suitable feedstock for future generation
area experiences a warm and humid climate with distinct
biodiesel. C. inophyllum is also regarded as a medicinal plant
rainy season. C. inophyllum seeds were collected during May-
in many parts of the world and almost all parts of the tree is
June 2013 from the mature trees with well developed crowns
used as medicine1 and it is a suitable species for coastal
in the coastal areas of Thirupuram Panchayath (805’ N and
afforestation1-2.
7609’ E), Thiruvananthapuram, Kerala. Seed lots were
The seed germination in C. inophyllum is found to be sporadic
thoroughly hand mixed to improve the homogeneity and graded
and it may take longer period to achieve maximum germination
into weight classes 2-5 (small), 5-8 (medium) and 8-11 g
due to hard pericarp2. Many pre-sowing treatments used
(large). The graded seeds were subjected to pre-sowing
worldwide to improve and synchronize the germination.
treatments viz. T1: Intact seed coat (Control), T2: Seed without
Grading of seeds based upon their size and weight is a
hard coat, T3: Alternate wetting and drying in cow dung slurry
common practice in a majority of plant species as it has been
for six days, T4: Soaking in cold water for 24 h and T5:
found to regulate the germination and subsequent seedling
Scarification (damage to seed coat by beating). The pre-
growth in many species3-5. Hence, the present study was
treated seeds belonging to respective weight classes were
undertaken to evaluate effect of different pre-sowing treatments
sown (100x 3) in polythene bags (11.43x15.24 cm, gauge 75
to hasten germination in the size graded C. inophyllum seeds
micron) containing the medium soil, sand and FYM in the
and to evaluate the influence of seed size variation on initial
ratio 9:3:1 and daily germination counts was recorded. At the
nursery performance of seedlings.
end of the trial, the percentage and Peak Value of germination
and Germination Value (GV) were calculated6. Since, the
treatment T2 (seeds without hard coat) produced the highest
NAAS Rating (2016)-4.20
 IMPACT OF SEED WEIGHT AND PRE-SOWING TREATMENTS
seed germination and others were low, the seedlings
subjected to T2 belonging to different weight classes were
grown in nursery and 12 seedlings per weight class were
destructively sampled at fortnight intervals till 60th day of sowing
to record height, collar girth, number of leaves, tap root length,
and total biomass (dried to constant weight in oven at
700C±20C). Another set of 12 seedlings were destructively
sampled simultaneously to record the assimilatory area (leaf
area) using an area meter (Model LI 3100 LI-COR, Nebraska,
USA). Growth analysis indices like specific leaf area (SLA),
leaf area ratio (LAR), leaf weight ratio (LWR), and Root : Shoot
ratio were calculated from leaf area and dry weights of
harvested plants. The following formulae were used to calculate
those parameters7.
LAR= Leaf area per plant / Plant dry weight, LWR= Leaf dry
weight/ Plant dry weight, SLA= Leaf area/ Leaf weight and
SLW= Leaf weight/ Leaf area. Root: shoot ratio of the seedlings
was calculated (Hunt 1990) by dividing the average value of
root weight by shoot weight. Relative growth rate (RGR)
was calculated7.
RGR = (log e W2 − log e W1 ) /(t 2 − t1 )
where, W1 and W2 are the dry weight determined at time t1
and t2 respectively and it was expressed in g g-1 month-1.
Net assimilation rate (NAR) is an index of the productive
efficiency of plant calculated in relation to the total leaf area.
It was calculated from the formula given by Hunt (1990):
NAR= (W2 −W1 )(loge LA2 − loge LA1) /(LA2 − LA1)(t2 − t1)
where, W2 and W1 are the dry weights at time t2 and t1 respec-
tively, LA2 and LA1 are the leaf area at time t2 and t1 respec-
tively and expressed in g cm-2 month-1
The physiological characters like chlorophyll content of the
seedlings were estimated8. The chlorophyll stability index of
the seedlings was estimated9.
Quality index which is a measure to assess the quality of
seedling based on the height, stem diameter and dry biomass
was also calculated10.
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Biovolume index, which is a non-destructive quick method to
calculate the above-ground portion of the tree seedlings was
calculated using the formula suggested10.
The vigour index (VI) of the seedlings was calculated using
the formula: VI = Germination Percentage x (Shoot length +
Root length or total seedling length)11.
The data on germination were subjected to two way analysis
of variance and the treatment means were compared with lsd
(least significant difference) wherever necessary. The seedling
growth, biomass production and growth analysis and quality
indices were subjected to one way repeated ANOVA.
RESULTS AND DISCUSSION
Two way analysis of variance revealed significant variation
(p=0.01) in germination percentage, Peak Value and
Germination Value due to interaction effect of weight class
and pre-treatment, whereas, the Mean daily germination
(MDG) did not vary significantly among various weight classes
or pre-treatment. With regard to germination percentage, the
highest value was (83.33±3.33 %) recorded for seeds
belonging to medium class devoid of hard seed coat (T2)
followed by large size class seeds devoid of hard seed coat
(T2) (76.87±3.004) (Table-1). The least germination occurred
in the medium class seeds subjected to alternate wetting
and drying in cow dung slurry (T3) and large seeds kept as
control T1 (10.0±2.0). Although, MDG of the seeds did not
vary statistically, highest MDG (1.78±0.83) was recorded for
seeds belonging to large size class, those were mechanically
scarified (T5) and the lowest (0.6±0.20) MDG observed in seeds
devoid of seed coat (T2) in belonging to medium weight class.
Peak value of germination, which is the speed of germination
was highest (0.36±0.06) for seeds belonging to large weight
class, which were mechanically scarified (T5) followed by that
devoid of seed coat in medium weight class (T2) (0.35±0.30).
Meanwhile, Peak Value of germination was the lowest for
large seeds kept as control (0.04±0.02). The Germination
Value which is the product of final MDG and Peak Value of
germination was the highest (0.61±0.18) for the seeds
belonging to large class, those were mechanically scarified
(T5). Lowest (0.01±0.04) Germination Values were recorded
 (431) KUMAR, AJEESH AND JIJEESH

. Table-1. The germination parameters of Calophyllum inophyllum as influenced by seed weight

and pre-treatment

Table-2. Growth attributes and biomass production of Calophyllum inophyllum as influenced by seed
weight at different fortnight intervals

for the seed belongs to large weight class kept as control
(T1) and that devoid of seed coat (T2) in medium weight class.
The lowest (0.10±0.05) Germination Values was recorded for
the seed belongs to large weight class which were
mechanically scarified (T5).
Effect of seed weight variation on seedling performance:
Analysis of variance revealed significant difference in collar
girth, number of lateral roots and tap root length of the seedlings
due seed weight over time, whereas the shoot height did not
vary significantly among various weight classes. With regards
seedling height, the highest value was (22.07±1.90 cm) recorded
for seedling belonging to large weight class (Table- 2). The
least (14.73±0.64 cm) height was occurred in seedlings
belonging to small weight class. Meanwhile, highest collar
 IMPACT OF SEED WEIGHT AND PRE-SOWING TREATMENTS (432)

Table-3. Physiological parameters of Calophyllum inophyllum as influenced by seed weight

at different fortnights

girth (2.53±0.31 cm) was recorded for large weight class
seedlings and the lowest value (1.83±0.72 cm) was observed
in medium class. Tap root length was highest (34.67±3.06
cm) in seedlings belonging to large size class and it was the
lowest in small weight class (21.07±1.51).The number of lateral
roots was the highest (34.67±3.06) for the seedlings belonging
to large class and lowest (21.07±1.51) value was recorded for
the seedling belongs to small weight class. The total biomass,
Root: Shoot and leaf area of the seedlings varied significantly
among different size classes and the highest values were
obtained for seedlings obtained from large weight class
followed by those of medium weight and small weight class
seeds (Table-2).
Values with similar alphabet with in row at different
fortnight are homogenous: Analysis of variance revealed
significant difference in Net Assimilation Rate (NAR), Relative
Growth Rate (RGR), Chlorophyll a & b, total chlorophyll content
and chlorophyll stability index due to seed weight at different
growth intervals. Whereas, the Leaf Area Ratio (LAR), Leaf
Weight Ratio (LWR), Specific Leaf Area (SLA) and Specific
Leaf Weight (SLW) did not vary significantly among various
weight classes over time.
Initially, the LAR was the highest in seedlings of small and
medium weight class. At the end of the study the large size
class seedlings recorded the highest LAR (77.19±13.44).
(433) KUMAR, AJEESH AND JIJEESH
Table-4. Seedling quality indices of Calophyllum inophyllum as influenced by size class in
different fortnights
Similar trend was observed in LWR. At the end of the study
period, the seedlings obtained from large weight class seeds
recorded the largest LWR (0.66±0.02). The values of SLW
(0.01±0.00), NAR (0.04±0.00), RGR (0.02±0.00), Chlorophyll
a (1.82±0.02), Chlorophyll b (0.44±0.02), total chlorophyll
content (2.37±0.07) and Chlorophyll Stability Index
(66.76±1.79) also were the highest in seedlings belonging to
large weight class (Table-3). Whereas, the seedlings obtained
from medium size class recorded the highest Specific Leaf
Area (120.05±12.88) at the end of the study period. The lowest
LAR (66.22±15.00), NAR (0.02±0.01), RGR (0.01±0.00),
Chlorophyll a (1.58±0.03), chlorophyll b (0.25±0.02), total
chlorophyll content (1.57±0.30) and Chlorophyll Stability Index
(42.91±2.25) were recorded for small weight class and a
inverse trend were obtained in the SLA (116.22±16.92) with
lowest values for large weight class.
Analysis of variance revealed significant difference in vigour
index, Quality index and Bio-volume index due to weight class
at different stages of growth. The vigour index was 1.8 fold
greater (4361.09±311.43) for the large weight class seedlings
compared to (2386.79±127.20) small weight class seedlings
(Table-4). The similar trends were observed in Quality index
and Bio-volume index. Greater than 100 per cent increase of
Bio-volume index was observed in large size class seedlings
compared to small weight class. The highest quality index
(0.77±0.04) and Bio-volume index (204.72±29.89) were
recorded for the seedlings belonging to large weight class
and lowest for small weight class at the end of the study
period (Table-4).
Various pre-sowing treatments influence the rate of seed
germination, enabling the period for germination to be reduced
from several months to only a few weeks. It is observed that
the germination of seeds in C. inophyllum is sporadic and it
may take longer period to achieve maximum germination. Our
own nursery experiences indicated a long period for germination
of C. inophyllum. This might be due to hard pericarp or
presence of seed dormancy. Different pre-sowing treatments
to overcome dormancy have been tried in this species. The
effects of chemical scarification, mechanical scarification, hot
water and cold water treatments on germination of seeds of C.
inophyllum and found that that mechanical scarification had
recorded the highest germination percentage, followed by cold
water treatment12. The positive influence of the pre-sowing
treatments on germination of C. inophyllum13. We studied the
effect of pre-sowing treatments on germination of size graded
C. ionophyllum seeds. Size grading based on weight
significantly influenced the germination with seed belonging
to medium class followed by large class recording a higher
germination. Of the pre-sowing treatments, T2 (seed without
hard coat) emerged as the best in all size classes. Earlier
workers had reported a higher germination of over 95 % for
the same treatment. A number of mechanisms have been
proposed by which the seed coat can impose dormancy on a
seed which include the mechanical restriction of germination
of the embryo, prevention of the exit of inhibitors from the
embryo, presence of chemical inhibitors of germination,
restriction of water uptake and restriction of oxygen uptake14.
Generally, bigger seeds germinate quicker and would take
 IMPACT OF SEED WEIGHT AND PRE-SOWING TREATMENTS (434)

lesser duration when compared to that of smaller ones15-21. In
the present investigation the number of days taken for
germination in T2 was 21 days, whereas, control germinated
after 72 days.
In the present study, the initial nursery performance of the
seedlings as measured by height, collar girth, tap root length,
number of lateral roots and total biomass varied significantly
due to seed weight over different periods22-23. There exist
variation in individual seed weight within a species and it can
vary from nearly constant to as high as 16-fold due to several
factors like genotype variation, competition for the limited
resources, environmental history to which mother plants are
exposed, and tradeoff between seed mass and number of
seeds per fruit. Seed traits such as seed size, seed coat
thickness, shape and moisture content are already known to
affect seed germination, dormancy and early seedling
establishment24-32. In the present study, also the higher seed
weight favoured the growth of the seedlings with the seedlings
belonging to larger size grade recording higher performance
followed by medium and small weight class. The various growth
analysis and physiological parameters as well as the seedling
quality indices of the seedlings belonging to various weight
classes were compared in the present study. For instance,
the LAR which expresses leafiness of the plant or the leaf
area formed per unit of biomass and LWR which expresses
the dry weight of leaves to total dry weight were initially higher
in small weight class seedlings, however towards the end of
the study period a higher LAR and LWR was recorded in the
large class seedlings. Specific leaf area (SLA) is a measure
of the leaf area of the plant to leaf dry weight varied with seed
size and at the end of the study; the medium seedlings
recorded a higher SLA indicating a higher photosynthesizing
area. Whereas, SLW which is a measure of leaf weight per
unit leaf area didn’t vary considerably among different size
grades. The positive relation between seed weight and
seedling growth in our study might be ascribed to the higher
leaf area for the seedlings of larger weight class. The larger
might be due to the higher number of root surface area for the
absorption of water and mineral nutrients and a higher leaf
area with high chlorophyll content for greater biomass
accumulation.
Physiological parameters like chlorophyll content and
chlorophyll stability index may be taken as an indication of
the seedlings performance. Chlorophyll stability index (CSI)
is an indication of the stress tolerance capacity of plants.
Literature on the effect of seed size on physiological
parameters of the C. ionophyllum seedlings is scanty. The
drupe size did not influence the chlorophyll content of Ceiba
pentandra seedlings35. Whereas, the higher chlorophyll content
in Tectona grandis seedlings33. Similar to them our study
also recorded a higher chlorophyll content and stability in
large size seedlings. The NAR and RGR were also higher in
the seedlings from large and medium weight class seeds. A
higher RGR is an index of faster seedling growth. Seedling
quality indices also varied among the weight classes. The
vigour index, quality index and Bio-volume index of the
seedlings was in the order large> medium> small.
From the study it can be conducted that seed weight and pre
treatments significantly influenced the seed germination of
Calophyllum inophyllum and the seeds belonging to large
weight class without hard seed coat (T2) produced the highest
germination. The seed weight also influenced the initial seedling
performance with large weight class seedlings recording higher
growth and biomass production which was supported by the
physiological and growth analysis indices of those seedlings.
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