Fifteen kenaf lines collected from kenaf and Jute Improvement Programme of Institute of Agricultural Research and Training (I.A.R.& T.) were subjected to digital imaging analysis using USB microscope with digital imaging software (Veho™ UK) and Vernier caliper to study the seed morphometric of available kenaf accession and the possibility of using the morphometric data to determine variations between the accessions. Ten seeds in four replicates of each seed lot were randomly selected and measurement of the seed length, seed width, seed angle and seed thickness were taken. The measurements were inputted and saved into Microsoft excel from where the mean value of each parameters were calculated for each replicates. Data were subjected to Analysis of variance, correlation analysis, principal component analysis and clustering analysis. Variation exit among seed of kenaf accessions though they had similar microscopic appearance features. Seed area, which was a function of seed length and seed width contributed largely to the variation that exist between the seed of kenaf accessions. Accession HC - 583 - 31 2 , clearly distinguished itself from others and therefore can be used in parent selection during breeding programmes. The inclusion of this seed morphometrics trait in taxonomic description of kenaf is recommended to increase the accuracy of morphological classification of kenaf.

1. Seed morphometric studies of some Kenaf (Hibiscus canabinus) accessions
ABSTRACT:
Fifteen kenaf lines collected from kenaf and Jute Improvement Programme
of Institute of Agricultural Research and Training (I.A.R.& T.) were subjected to digital
imaging analysis using USB microscope with digital imaging software (Veho™ UK) and
Vernier caliper to study the seed morphometric of available kenaf accession and the
possibility of using the morphometric data to determine variations between the
accessions. Ten seeds in four replicates of each seed lot were randomly selected and
measurement of the seed length, seed width, seed angle and seed thickness were
taken. The measurements were inputted and saved into Microsoft excel from where
the mean value of each parameters were calculated for each replicates. Data were
subjected to Analysis of variance, correlation analysis, principal component analysis
and clustering analysis. Variation exit among seed of kenaf accessions though they
had similar microscopic appearance features. Seed area, which was a function of seed
length and seed width contributed largely to the variation that exist between the seed
of kenaf accessions. Accession HC-583-312
, clearly distinguished itself from others and
therefore can be used in parent selection during breeding programmes. The inclusion
of this seed morphometrics trait in taxonomic description of kenaf is recommended to
increase the accuracy of morphological classification of kenaf.
029-033 | JRA | 2012 | Vol 1 | No 1
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www.jagri.info
Journal of Research in
Agriculture
An International Scientific
Research Journal
Authors:
Adetumbi JA1
,
Adekoya MA1
,
Adeyeye OO1
and
Ogunbiyi B2
.
Institution:
1. Institute of Agricultural
Research and Training,
Obafemi Awolowo
University, P.M.B.5029,
Moor Plantation. Ibadan
Nigeria.
2. Federal College of
Agriculture, P.M.B.5029,
Moor Plantation, Ibadan.
Corresponding author:
Adetumbi JA
Email:
jaadetumbi@yahoo.com.
Web Address:
http://www.jagri.info
documents/AG0008.pdf.
Dates:
Received: 20 Nov 2011 Accepted: 13 Dec 2011 Published: 25 Jan 2012
Article Citation:
Adetumbi JA, Adekoya MA,Adeyeye OO and Ogunbiyi B.
Seed morphometric studies of some Kenaf (Hibiscus canabinus) accessions.
Journal of Research in Agriculture (2012) 1: 029-033
Original Research
Journal of Research in Agriculture
JournalofResearchinAgriculture An International Scientific Research Journal

2. INTRODUCTION
Cultivar identification for the attainment of
Plant Breeding Rights (PBR) requires taxonomic
and genetic approach to determine cultivar
distinctness. The most common methods used for
varietal identification are morphological
characterization which are often time consuming,
expensive, influenced by environment and therefore
lack precision. Morphometric variation has been
widely used in determining potential plant diversity
at the visible level in the sea grass Halodule
wrightii (Creed, 1997). Wyllie-echeverria et al.,
(2003) have also proved that metric measurements
on seeds can be very important in quantitative
variables for determining size and shape of seeds.
Kenaf is a member of the hibiscus family (Hibiscus
cannabinus.L) related to cotton and okra. It is an
annual or biennial herbaceous plant growing to 1.5
– 3.5m tall with a woody base. Kenaf are used by
small pulp mills primarily in countries like China,
India and Thailand. (Dempsey 1975). In Nigeria,
main uses of kenaf fibre are producing rope, twines,
coarse cloth (similar to that made from jute), and
paper. Other uses of kenaf fibre include engineered
wood insulation, various types of environmental
mats, packing materials and material that absorbs
oil and liquids. (Mabberly, 1987) Many varieties of
kenaf are often mentioned in literature based on
their performance in one character or the other but
concise varietal identification of kenaf is always
problematic and knowledge on genetic diversity of
kenaf varieties is limited. Since all forms of
breeder’s protection require some measure of
distinctness, cultivars identification is therefore a
pre- requisite for the effective provision of Plant
Breeding Rights (PBR). It is based on these facts
that this experiment set to study the seed
morphometric of available kenaf accession and the
possibility of using the morphometric data to
determine variations between the accessions.
MATERIALS AND METHODS:
Clean seed of fifteen kenaf lines (Table 1)
were collected from kenaf and Jute Improvement
Programme of Institute of Agricultural Research
and Training (I.A.R. & T). Ten seeds in four
replicates were randomly selected from each seed
lot of the 15 accessions, thereafter, each of the ten
seed samples were subjected to digital imaging by
placing each seed sample under the USB
microscope in such a way that the flat embryo axis
of the seed was facing the lens of the camera under
light and 40X magnification. The light and
magnification focus adjuster on the USB
microscope was adjusted until desired brightness
and magnification that gives clear image. The
seeds were then viewed on computer system before
the image of the seed was captured. The captured
image was then double clicked to be previewed and
displayed at the measurement window of the USB
microscope. At the display of the seed image,
magnification at which the seed image was captured
was inputted; thereafter measuring tools were used
to take various precise measurements of the features
on the seed in millimeters. Measurements taken
were:
i. Seed length - This is the greatest dimension on
the seed taken at base of the seed.
ii. Seed width – This is the cross section taken at
right angle to the length of the seed.
iii. Seed angle – This is the angle formed from the
tip of the seed to the base.
iv. Seed area – This is calculated from the seed
length and seed width using the formula: ½
base x heights.
v. Seed thickness was however obtained by
holding each kenaf seed in between the vernier
caliper to digitally measuring the thickness
The values of these parameters taken were
saved in Microsoft excel from where the mean
value of each parameters were obtained for each
replicates. Data collected were subjected to analysis
of variance and correlation analysis using SPSS.
Principal component procedure was used to
estimate the variability among the accessions while
the clustering analysis was used to group the
accessions based on variables with highest variation
based on morpho-metric similarities.
030 Journal of Research in Agriculture (2012) 1: 029-033
Adetumbi et al.,2012
Entry number Accessions
1 Cuba 108
2 Exshika24 4
3 Ifekan 100
4 AU-71-93
5 HC-583-312
6 AU-75- 413
7 V1-100-10 1
8 AC-313-302
9 2QQ-171
10 G45-23
11 AC-313-292
12 PURPLE FLOWER
13 AC-313-244
14 A-60 282-5
15 2QQ-13
Table 1: List of Kenaf Accessions used for
the experiment

3. RESULTS AND DISCUSSIONS
Variability among metric measurements on
kenaf seed
Mean squares from analysis of variance of
seed morphometrics parameters for fifteen kenaf
lines revealed significant differences at P<0.05 in
all the parameters except seed angle (Table 2).
From the result, it is clear that variation exist
among the seeds of fifteen kenaf accessions with
respect to some of the characters evaluated. This
indicates that descriptors of seed differences can be
effectively exploited in physiological studies as
stated by Dell’ Aquila (2004), and in cultivar and
genotype description.
Mean values of the morphometric measurements
of kenaf seed.
Mean morphometric data of fifteen kenaf
accessions are presented in Table 3. 2QQ-13
had
the longest seed length (5.42 mm). The shortest
seed length is recorded for HC-583-312
(4.15mm)
was significantly different from other accessions at
P<0.05. Similarly, HC-583-312
recorded the
shortest seed width (3.23mm) and seed area
(6.72mm), which were also significantly different
from other accessions. Thickness of Ex-Shika 24-4
recorded the highest value (2.51mm) which was not
significantly different from another eight
accessions. Also, Purple flower accession had the
lowest thickness value (2.03mm), which was not
significantly different from AU2452-4A
. There was
no significant difference in the seed angle of almost
all the accessions. Cuba 108 recorded the highest
seed angle (75.66 mm) which was not significantly
different from the value recorded for AU-75-413
(75.60mm) while AC-313-302 recorded
significantly lowest seed angle (69.81mm).
Intercharacter relationship among kenaf seed
Correlation coefficients among the
mormhometric data of the fifteen kenaf accessions
shows that seed width significantly and positively
correlated with seed length and seed area (r =
0.0738 and r = 0.0939 respectively).Table 4). Also
significant positive correlation exist between Seed
length and seed area (r =. 0.039). Though the
correlation between seed thickness with both seed
width and seed length are positive, they are not
significant at P<0.05. Seed angle is negatively
correlated with seed width and seed thickness and
they are not significant.
Journal of Research in Agriculture (2012) 1: 029-033 031
Adetumbi et al.,2012
Sv Df Seed length Seed width Seed thickness Seed angle Seed area
Accessions 14 0.40* 0.23* 0.06* 11.23 5.10*
Error 45 0.04 0.04 0.02 8.63 0.53
Total 59
Table 2: Mean square values for kenaf seed morphometrics.
Accessions Seed length Seed width Seed thickness Seed area Seed angle
Cuba 108 5.00bcdef 3.61cde 2.33abc 9.02def 75.66a
Exshika24 4 5.22abc 3.80abc 2.51a 9.97abcd 71.50ab
Ifeken 100 4.94cdef 3.61cde 2.39ab 8.90def 71.84ab
AU-71-93
5.15abcd 4.02ab 2,41ab 10.36abc 72.37ab
HC-583-312
4.15g 3.23f 2.29bcd 6.72g 71.50ab
AU2452 4A
4.70f 3.40ef 2.11de 7.80f 73.34ab
V1-100-10 1 4.72ef 3.46def 2.35ab 8.17ef 73.55ab
AC-313-302
5.18abcd 3.94abc 2.32abcd 10.19abc 69.89b
2QQ171 5.03bcdef 3.78abcd 2.27bcd 9.49bcd 71.99ab
G4523 4.80ef 3.70bcd 2.37ab 8.87def 70.99ab
CUBA OVATE 671 5.04bcde 3.66cde 2.28bcd 9.22cde 72.98ab
PURPLE FLOWER 4.88def 3.62cde 2.03e 8.81def 71.46ab
AC-313-244 5.28ab 4.00ab 2.32abcd 10.57ab 72.78ab
A-60 282-5 5.31ab 4.06a 2.34abc 10.79a 74.69ab
2QQ-13
5.42a 3.85abc 2.13cde 10.45ab 75.60a
Table 3: Means of Kenaf seed morphometrics
Mean followed by the same alphabet within the same column are not significantly different from each other.

4. The positive and significant correlation that
exists between seed width and the other two
characters (seed length and seed area) indicates that
improving on any of the parameter will
significantly increase other characters. Also, since
they are significantly correlated, each can be used
as substitute in any evaluation and classification
studies and as such these characters possessed
greater practical value for seed discrimination in
kenaf. Wyllie-echeverria et al. (2003) did show that
seed size and seed metrics were important
discriminators of Zostera marina. Similar result
was obtained on Jathropa by Kaushik et al.,(2007).
Seed angle had negative and non significant
relationship with seed width and seed thickness.
This denotes that as the width of the seed increases,
the angle reduces or improvement on the seed width
or thickness will significantly reduce the seed
angle, although the effect might not be significant.
Factors contributing to variations among kenaf
seeds
The result of the principal component
analysis showing the Eigen values and percentage
contribution are presented in Table 5. Principal
component 1 and 2 are of major importance. The
result shows that PC 1 and PC 2 accounted for
59.80% and 24.51% variance respectively.
Principal Component 1 was loaded largely by seed
area (0.994) indicating that seed area contributes the
largest variability while seed angle contributes the
least variability (0.229). However, with PC 2, seed
angle (0.799) contributes the largest variability
while seed width (-0.119) contributes the least
variability. The eigen values of the two principal
component are more than one.
The output of the principal component
analysis shows that different characters contributed
differently to the total variation as indicated by their
eigen values as well as loading on different
principal axis. Seed area contributed largely to the
variation that exists in PC 1 because the seed area is
the function of other two variables (seed length and
seed width). However, in PC 2, seed angle
contributed the largest to the variation that exist
between the accessions because the seed angle is
independent of other parameters
Degree of relationship among kenaf accessions
based on seed metrics
The result in Figure 1 shows the dendogram
032 Journal of Research in Agriculture (2012) 1: 029-033
Adetumbi et al.,2012
Table 4: Correlation coefficients of observed kenaf
seed mormphometrics
Seed
width
Seed
length
Seed
thickness
Seed
angle
Seed
area
Seed
width
1 0.738* 0.148* -0.102 0.939*
Seed
length
1 0.104 0.244 0.923*
Seed
thickness
1 -0.072 0.145
Seed
angle
1 0.069
Seed
area
1
Character PC 1 PC 2
Seed width 0.965 -0.119
Seed length 0.969 0.142
Seed thickness 0.284 -0.743
Seed angle 0.229 0.799
Seed area 0.984 0.005
Eigen value 2.99 1.23
% variance 59.79 24.51
Table 5: Principal Component analysis showing the
eigen value and percentage contributions of each
variable.
Case 1 Cuba 108 Case 9 2QQ171
Case 2 Exshika24 4 Case 10 G4523
Case 3 Ifekan 100 Case 11
CUBA OVATE
671
Case 4 AU-71-93
Case 12
PURPLE
FLOWER
Case 5 HC-583-312
Case 13 AC-313-244
Case 6 AU2452 4A
Case 14 A-60 282-5
Case 7 V1-100-10 1 Case 15 2QQ-13
Case 8 AC-313-302
Legend:
Figure 1: Dendogram showing the morphometric
similarity among fifteen kenaf accessions
Cluster Tree

5. from single linkage cluster analysis (SCLA) of the
fifteen Kenaf accessions. At a minimum distance of
0.00 level of similarity, all the accessions were
distinct from each other while at a distance of 1.00
level of similarity, only accession five (HC-583-
312
) can be distinguished from other accessions that
formed into a cluster.. However, at 0.68 level of
similarity, accessions Cuba 108(1), 2QQ-13
(15) and
A-60-282-5(14) were similar to one another but
clearly distinct from other accessions.
The dendogram generated from similarity
and distance matrices provided an overall pattern of
variation as well as degree of relationship among
the accessions. The result from this study has
provided two major clusters at the 1.00 level of
similarity, with fourteen out of the fifteen
accessions forming a cluster. This indicates that
only one accession (HC-583-312
) has distinct
features that can be used to distinguish it from
others. However, at a lesser level of similarity
(0.68), three clusters can be identified. It is
therefore clearly shown from this study that seed
morphometrics of kenaf could not provide
convincing discriminatory evidence in the
classification of kenaf genotypes.
CONCLUSION
This experiment has shown that this imaging
technique can be applied to the identification of
seeds and it has shown that seed of kenaf varieties
had similar microscopic appearance features. The
seed morphology of the kenaf varieties however
proved that there are differences in their length,
width and thickness. However, this distinction
cannot be used as a tool to identify kenaf varieties.
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Dempsey JM. 1975. Fibre crop. Univ of Florida
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Kaushik N, Kumar K, Kumar S, Kaushik N, and
Roy S. 2007. Genetic variability and divergence
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Journal of Research in Agriculture (2012) 1: 029-033 033
Adetumbi et al.,2012
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