Bio-Research Vol.21 No.1
pp.2003-2018 (2023)
2003
Original Research
Article
Journal of Biological
Research
& Biotechnology
Bio-Research Vol. 21 No.2; pp.
2003-2018 (2023). ISSN (print):1596-7409; eISSN (online):2705-3822
Effects of indole butyric acid
and coconut liquid endosperm
on rooting of Crateva adansonii
stem
1
Nwankwo Maryrose Chigozie,
§1
Ogwo Nnamdi,
1
Oti Valentine Ogbonna,
4
Nweze
Nkechinyere Onyekwere,
1
Ohabuiro Augustina
Akudo,
2,3
Ene Onuora Chikezie
1
Department of Plant Science and
Biotechnology, Michael Okpara University of Agriculture,
Umudike
Nigeria
2
Department of Agriculture Alex
Ekwueme Federal University Ndufu-Alike, Abakaliki
Nigeria
3
Department of Horticulture and
Plant science Jimma University, Jimma Ethopia
4
Department of Plant Science and
Biotechnology, University of Nigeria, Nsukka, Nigeria
§
Corresponding author: Ogwo
Nnamdi. Email., nnamdiogwo1@gmail.com
Abstract
Indole butyric acid (IBA) and
coconut liquid endosperm (CLE), two rooting agents, were tested in
three
concentrations to see how they
affected the rooting of Crateva adansonii DC stem cuttings. 300 mg/L,
200
mg/L, and 100 mg/L (IBA) of
rooting material and 100%, 80%, and 60% of CLE were the
concentrations
that were assessed. This was done
in three replications using completely randomized design (CRD).
The
setup, which lasted six months,
was on exhibit in the Botanic Garden at the Department of Plant
Science
and Biotechnology, University of
Nigeria, Nsukka. While control stem cuttings took the longest to start
and
finish bud break on the stem
cuttings, higher concentrations of the rooting chemical substances, 300
mg/L
IBA and 100 percent CLE
respectively, influenced the shortest times (initial and final). The
results showed
that the control had the fewest
buds that sprouted into leaves, whereas 300 mg/L IBA and 100
percent
CLE had the highest percentage of
buds that turned into leaves on the stem cuttings. Regardless of
the
treatment concentrations
utilized, a thin callus formed on the stem cuttings. Observations on
rooting
revealed that not all callused
stem cuttings subsequently took root. IBA and CLE were applied to
stem
cuttings in various
concentrations, and the results were seedlings with lateral and feeding
roots. The
maximum percentage of rooting
response was seen in stem cuttings treated with 300 mg/L IBA and
100
percent CLE, respectively, while
the control did not root. The findings of measuring the aerial and
sub
aerial growth characteristics of
seedlings revealed that 300 mg/L IBA and 100 percent CLE had a
greater
impact on the development of
higher values of the growth parameters examined than cuttings treated
with
lower concentrations of the two
rooting agents. Based on the findings of this study, it is possible to
draw
the conclusion that C. adansonii
can be multiplied vegetatively by using rooting agents to help
stem
cuttings take root. It is
recommended that more research be done using rooting medium with a
higher
concentration and rooting agents
like auxins at different concentrations.
Keywords: Crateva adansonii,
Indole buteric acid (IBA), Coconut liquid endosperm (CLE), Rooting and
callus
Received November 28,2022;
Revised June 10, 2023; Accepted June 21, 2023
https://dx.doi.org/10.4314/br.v21i2.7
This is an Open Access article distributed under the terms of the
Creative Commons License [CC
BY-NC-ND 4.0] http://creativecommons.org/licenses/by-nc-nd/4.0.
Journal
Homepage:
http://www.bioresearch.com.ng.
Publisher: Faculty of Biological
Sciences, University of Nigeria, Nsukka, Nigeria.
Bio-Research
Vol.21 No.1 pp.2003-2018 (2023)
2004
INTRODUCTION
There are roughly 70 species in
the genus
Crataeva (Capparaceae), most of
which are
found in the tropics and
subtropics (Kumar et
al., 2020). One of these, Crateva
adonsinii,
exhibits remarkable health
advantages. It is a
medium-sized deciduous tree that
bears the
name of the Greek botanist
Cratevas (Kumar et
al., 2020). Due to poor seed
germination and
poor seedling establishment, this
tree is rarely
widely planted (Igoli et al.,
2014). In nature, the
tree only generates a small
number of root
suckers, which limits its range
(Kher et al.,
2016). In the savanna and forest
regions, it
grows as a little tree and is
extensively
dispersed. They are frequently
observed along
riverbanks throughout Africa and
resemble the
Asian Crateva religoasa (Igoli et
al., 2014).
Although Crateva adansonii is
consumed as
food and is significant in
medicine and
commerce, very little is known
about its
nutritional worth. According to
Agbankpe et al.
(2015), who established their
nutritional
benefits, they are high in
calcium, zinc, and
proteins. When it comes to
treating ear
infections, the leaves are highly
sought after in
Eastern Nigeria, whilst the
Yoruba ethnic group
in Western Nigeria uses the same
leaves as a
mild headache counter irritant
(Agbankpe et al.,
2015). Rubefacient substances
used on cysts
include the ground leaves and
bark (Kabore et
al., 2015). The bark of the plant
has been used
to cure ailments like syphilis,
jaundice, yellow
fever, mycotic infections, and to
hasten the
healing of wounds, according to
Ajali et al.
(2010), Adjabga et al. (2017),
and Kumar et al.
(2020). The long list of
successes recorded
from the use of the extracts of
the various
tissues of the species in
treatment of different
ailments and hence, the need to
make available
large quantity of the plant
materials, which can
sustain biotechnological
exploitation excited
interest in selecting rooting of
the stem cuttings
which is an aspect of vegetative
propagation for
this study.
Among all seedling multiplication
approaches,
rooting of stem cuttings has
become the fastest
method used to produce large
quantity of
uniform seedlings. It contributes
significantly to
the conservation of the species
both for present
as well as future
biotechnological exploitation of
its medicinal properties and
sufficiently ensures
their availability for research
activities
(Yeshiwas et al., 2015). Above
all, the present
study will provide baseline
information on the
methods for the vegetative
propagation of the
species, and suggest which can be
adopted by
local nursery men and farmers for
best result.
Although, some researchers have
attempted
studying the impact of IBA on the
propagation
of plants, such as Bambusa
arundinacea
(Venkatachalam et al., 2015),
Rosa rubiginosa
(Yeshiwas et al., 2015), Duranta
erecta (Shiri et
al., 2019), Ocimum gratissimum
(scent leaf)
and Pterocarpus mildebraedii
(Okafor et al.,
2020), and Cornus sericea (Inoue
et al., 2022),
there is paucity of information
on the possible
application of IBA and CLE in
propagating
Crateva adansonii. Several
reports have shown
that seedlings produced by seeds
take longer
time to yield, probably because
of the effect of
prolonged seed dormancy, hence,
delaying
biotechnological exploitation of
their resources
(Nzekwe, 2006; Igoli et al.,
2014; Adjagba et al.,
2017). Based on insufficient
knowledge on how
to solve the problem of delayed
seed
germination of Crateva adansonii,
coupled with
numerous untapped health benefits
derivable
from its seeds, leaves, wood,
root and bark
which of course has skyrocketed
its demands,
it has become necessary to
develop other
methods of propagating this
important plant
species so as to enable full
exploitation of its
medicinal attributes. The
efficiency of two
rooting agents on the rooting and
seedling
development of mature stem
cuttings of
Crateva adansonii DC
(Capparaceae) was thus
compared.
MATERIALS AND METHODS
Site description for the
experiment
The research was done in the
botanic garden of
the University of Nigeria, Nsukka
Department of
Plant Science and Biotechnology,
during the
rainy season under cover of
shade. Nsukka is
located on latitude 6
o
51’56” N, longitude
7
o
24’22” E and 1,410ft above sea
level. The
approach involved planting of the
stem cuttings
in poly pot and mechanically
irrigating the
setup, avoiding the use of mist
drip irrigation or
bottom heat technique (Griffith,
1940). The
Centre for Basic Space Science at
the
University of Nigeria, Nsukka,
provided
meteorological information such
as average
monthly rainfall, temperature,
and humidity
during the course of the
study.
Materials description
Mature coconut fruits (Ghana
variety, because
of its high liquid endosperm
content) bought
from local market (Nsukka main
market) were
extracted of the liquid endosperm
content.
Bio-Research Vol.21 No.1
pp.2003-2018 (2023)
2005
Indole-3-butyric acid and
branches of Crateva
adansonii trees were obtained
from the
departmental laboratory and
botanic garden,
respectively. Uniform stem
cuttings were
derived from the Crateva
adansonii branches.
Top soil and poultry droppings
were collected
from uncultivated land from the
departmental
botanic garden and poultry farm
of the Faculty
of Agriculture,
respectively.
Plate 1: the image of a typical
Crateva adansonii plant.
Preparation of media and
rooting
substances
The growing substrate was
prepared from a 2:1
mixture of top soil and poultry
manure by
thoroughly mixing 40kg of the
former with 20kg
of the latter. Polypots measuring
(12 x 25 cm)
in size were used for the
experiment. The
polypots which contained 0.25kg
of the media
were perforated at the base and
sides to allow
for adequate drainage of excess
moisture. The
choice of the growing medium
which is
otherwise referred to as the
amended media
was based on the reported
qualities of
mixed/amended media over single
medium.
Nzekwe (2006) projected such
media as the
best being well-aerated and could
retain
enough moisture for plant growth
over a given
period.
The concentrations of IBA (SimSon
Chemtech,
India) used were prepared by
dissolving it into
three levels viz;300mg, 200mg and
100mg
separately in one litre of
distilled water. Coconut
liquid endosperm 100ml (100%) was
the
undiluted stock, 80 ml was
diluted with 20ml
water to get 80%, this was
followed by dilution
of 60 ml with 40ml water to get
60%.
Preparation of stem
cuttings
Defoliated C. adansonii branches,
each of
which carried 4-6 buds, were
trimmed to length
(30 cm) from trees in the
Department's botanic
garden. 42 stem cuttings of
C.adansonii in total,
divided into 3 replicates, were
utilized for the
experiment's 7 treatments (3
levels of IBA, 3
levels of coconut liquid
endosperm, and control)
(2 cuttings per treatment x 7
treatments x 3
replicates).
Before planting, each cutting lot
was tied with
twine and submerged for an hour
in the
appropriate rooting chemical
substance
concentration (300mg/L, 200mg/L,
100mg/L
IBA, and 100%, 80%, and 60%
CLE).
According to Hartmann and
Kerster's (2002)
advice, the upper cut end of the
stem cuttings
was sealed with paraffin wax
before planting to
reduce water loss, particularly
from the pith.
Cuttings that had not been bathed
in rooting
agent served as the control for
the experiment.
Bio-Research
Vol.21 No.1 pp.2003-2018 (2023)
2006
Stem cuttings were planted in a
slanting
orientation, one cutting per
polypot.
Experimental design
Complete randomization design
(CRD) was
used in the experimental design.
The setup
was on exhibit in a shaded area
of the
Department's botanic garden,
which received
daily mechanical irrigation and
manual weed
removal. Observations were made
on sign of
bud breaking which represented
growth
activities on the
cuttings.
Data collection
Daily checks were done to see if
the stem
cuttings' metabolic processes
started up again
during each watering cycle. The
parameters
tracked included initial and
final bud sprouting
times, the quantity of sprouted
buds produced
by each stem cutting, and the
times when buds
developed into
photosynthesis-producing
leaves. After the development of
the leaves,
stem cuttings were randomly
selected at four-
week intervals to examine callus
and root
development. Cuttings that had
developed
calluses and roots were labeled
and replanted
following each sampling.
Aerial growth and rooting
parameters
The following aerial growth
parameters such
as: number of sprouted buds on
stem cutting
with emphasis on the times of the
first and last
bud sprouting as well as the
quantity of buds
that developed into leaves were
recorded
likewise, the following rooting
parameters such
as: number of callused and rooted
stem
cuttings, type of callus
developed, types and
length of five longest roots in
each treatment
were also observed and recorded.
For each
parameter, mean of ten stem
cuttings were
determined. Leaf area was
estimated by adding
a constant to the product of the
leaf’s length and
width, whereas using a meter
rule, the root
length was calculated and
recorded in
centimeters, the rest parameters
were counted.
Data analysis
Using SPSS, the data were
subjected to an
analysis of variance (ANOVA), and
Duncan's
New Multiple Range Test (DNMRT)
was used
to differentiate the means at P
≤0.05. Plates,
figures, and tables are used to
show pertinent
data and observations.
RESULTS
Meteorological results comprising
of
rainfall, relative humidity and
temperature
The result for the mean monthly
rainfall
throughout the duration of the
trial is presented
in Figure 1. The results showed
that mean
monthly rainfall ranged from 120
to
440±12.73mm, with
September
(440±12.73mm) as the wettest
month. This was
followed by October (400±12.73mm)
while the
least was recorded in August
(120±12.73mm).
The mean monthly relative
humidity as
presented in Figure 2 showed that
relative
humidity ranged from 48 to
75±9.64% and that
the most humid month was recorded
in
September (75±9.64%) also. This
was followed
by October (70±9.64%) and July
(60±9.64%),
while August (48±9.64%) was the
least. The
results of the determination of
mean monthly
temperature are summarized in
Figure 3. There
were variations in the mean
monthly
temperature. The results showed
that the mean
monthly temperature had a range
of
28.4±0.29
o
C to 29.8±0.29
o
C and the hottest
month was August
(29.8±0.29
o
C). This was
followed by May
(29.6±0.29
o
C) and October
(29.6±0.29
o
C). The months of July
and
September (28.4±0.29
o
C) each had the coolest
temperature
comparatively.
Periods of bud breakage (initial
and final)
The results to determine the
initial and final
period of bud breakage on the
stem cuttings are
presented in Table 1. The
outcomes revealed
variation among the varying
levels of each
treatment in both bud breakage
periods.
Irrespective of rooting substance
and their
concentrations all the stem
cuttings including
the control (untreated stem
cuttings) had bud
flush as indicated in Plates 2a
and b. The initial
period of bud break ranged from
25.50±0.65 to
37.50±0.65 days after planting
(DAP). Stem
cuttings that have been treated
with 300mg/L of
IBA had the earliest period of
bud break
(25.50±0.65) DAP. It was followed
by cuttings
of stems that have been treated
with 100%
coconut liquid endosperm
(26.50±0.65) DAP
while the control took the
longest period
(37.50±0.65) DAP to experience
sprouting. This
make stem cuttings treated with
IBA to have
taken shorter period (25.50±
0.65) DAP to
sprout comparatively. Sprouting
observed in
stem cuttings treated with CLE
varied from
26.50±0.65 to 37.50±0.65
DAP.
Bio-Research Vol.20 No.3
pp.2003-2018 (2022)
2007
Figure 1:Mean monthly rainfall
variation
Figure 2:The average monthly
change in relative humidity
Figure 3: Mean monthly
temperature variation
Bio-Research
Vol.20 No.3 pp.2003-2018 (2022)
2008
The period of finial bud break
(termination of
bud sprouting) ranged from
33.75±0.48 to
48.75±0.85 DAP. The stem cuttings
treated
with 300mg/L IBA were also the
earliest to
terminate bud sprouting which was
recorded
from 33.75±0.48 to 42.00±0.91days
after
planting whereas the controls
took the longest
periods to terminate their bud
sprouting
(47.50±1.04 to 48.75±0.85)
DAP.
Comparatively, the initial period
DAP to bud
sprout termination at stem
cuttings treated with
300mg/L of IBA was the shortest
at 25.50±0.65
DAP. In CLE, 100%-treated stem
cuttings had
the shortest beginning time
(26.50±0.65 DAP),
whereas 60%-treated stem cuttings
had the
longest (37.50±0.65
DAP).
Percentage bud breakage
The effect of rooting substance
on percentage
of bud breakage at the end of bud
sprouting is
presented in Table 2. The results
showed
variation among the treatment
means and the
values ranged from 67.25±0.85%
to
88.50±0.65%. Stem cuttings
immersed in 200
mg/L of IBA produced the maximum
bud sprout
(88.50±0.65%), followed by those
treated with
100 percent CLE (88.00±0.71%).
Stem cuttings
that had been bathed in 300mg/L
of IBA
(83.50±0.65%) came next. The
results also
demonstrated that the control
(67.25± 0.85 %)
and stem cuttings soaked in 60
percent CLE
had the least. As seen in Plates
2a and b,
several cuttings treated with
Indole Butyric Acid
(IBA) and Coconut Liquid
Endosperm (CLE)
had multiple buds flush.
Comparatively, bud
break percentage was higher in
stem cuttings
soaked in three different
concentrations of IBA
than it was in three different
concentrations of
CLE.
Percentage of sprouting buds that
grew into
leaves
The percentage of buds that
eventually turned
into photosynthesis leaves is
shown in Table 3.
According to the findings, not
all of the buds that
emerged on the stem cuttings
survived and
turned into leaves (Plate 3).
Between
59.62±1.07% to 84.25±2.17% of
sprouting buds
turned into leaves. The buds that
developed on
stem cuttings treated with 100
percent CLE had
the maximum leaf development,
measuring
84.25±2.17%. Buds that emerged on
stem
cuttings treated with 300mg/L and
200mg/L IBA
had values of (82.75±1.11%) and
(79.75±0.85%), respectively. The
next to
appear on stem cuttings was from
the control,
which had a bud development into
leaves
percentage of 59.62±1.07%., this
was the
lowest.
Comparatively, the findings
demonstrated that
stem cuttings treated with CLE
had lower
percentages of buds developing
into leaves
than those that sprouted on stem
cuttings
treated with the three dosages of
IBA.
Additionally, regardless of the
rooting chemical
concentration, the data revealed
that roughly
25% of the stem cuttings had
numerous buds
flush that turned into leaves
(Plate 4).
Development of Callus on the stem
cuttings
of Crateva adansonii as influence
by IBA
and CLE
The observations made on the stem
cuttings'
basal region one month after the
growth of the
leaves revealed that not all of
the randomly
picked stem cuttings had
calluses. Additionally,
the stem cuttings generated light
calluses, as
seen in plates 5a and b,
regardless of the type
of rooting material and its
concentration.
Additionally, stem cuttings of
the control that
survived did not exhibit callus
development,
regardless of the growth of
leaves. Further
investigation revealed that some
callused stem
cuttings had sprouted among the
cuttings
treated with coconut liquid
endosperm and
indole butyric acid (IBA)
(CLE).
Callus percentage affected by
Indole Butyric
Acid (IBA) and Coconut Liquid
Endosperm
(CLE)
The percentage callus production
ranged from
28.00±2.16% to 81.75±1.18%,
according to the
results shown in Table 4. On the
stem cuttings
treated with 300mg/L of IBA, the
highest
percentage callus production,
81.75±1.18 %
was noted. Cuttings treated with
100% CLE
came in second (81.25±1.49%),
while those
treated with 60% of a similar
chemical had the
lowest value (28.00±2.16%).
Comparatively,
the results showed that
CLE-treated stem
cuttings exhibited lower
percentage callus
formation than those treated with
the three
levels of IBA concentrations. The
outcomes
also demonstrated that greater
concentrations
of CLE (100%) and IBA (300 mg/L)
significantly
influenced very high percentage
callus
formation on the stem cuttings,
with respective
values of 81.25±1.49% and
81.75±1.18%.
Bio-Research Vol.20 No.3
pp.2003-2018 (2022)
2009
Table 1: Rooting agents Indole
Butyric Acid (IBA) and Coconut Liquid Endosperm (CLE) effects on the
times of bud breakage on C.
adansonii stem cuttings
Periods of bud breakage
(Days)
Treatments
Initial (DAP)
Final (DAP)
IBA (mg/L)
100
34.50 ± 1.04
ab
42.00 ± 0.91
b
200
33.50 ± 1.04
b
38.50 ± 0.65
c
300
25.50 ± 0.65
c
33.75 ± 0.48
d
Control
37.00 ± 0.91
a
48.75 ± 0.85
a
LSD(𝑃 < 0.05)
2.85
2.29
CLE (%)
60
37.50 ± 0.65
a
46.00 ± 1.08
a
80
30.50 ± 0.65
b
38.50 ± 0.65
b
100
26.50 ± 0.65
c
35.75 ± 0.85
b
Control
37.50 ± 0.65
a
47.50 ± 1.04
a
LSD(𝑃 < 0.05)
1.99
2.84
In the vertical column, values
with the same alphabet(s) do not significantly differ from one another
(𝑃 <
0.05).
Table 2: Coconut liquid endosperm
(CLE) and indole butyric acid (IBA) effects on the
percentage of bud breaking on
Crateva adansonii stem cuttings
Treatments
Bud breakage (%)
IBA (mg/L)
100
83.50 ± 0.65
b
200
88.50 ± 0.65
a
300
83.50 ± 0.65
b
Control
78.50 ± 0.65
c
LSD (P<0.05)
1.98
CLE (%)
60
67.25 ± 0.85
c
80
76.50 ± 0.65
b
100
88.00 ± 0.71
a
Control
67.25 ± 0.85
c
LSD (P<0.05)
2.35
Values in the vertical column
that share the same alphabet(s) do not significantly differ
from one another (P
0.05).
Bio-Research Vol.20 No.3
pp.2003-2018 (2022)
2010
Plates 2a:Indole butyric acid
influences the bud
sprouts on C. adansonii stem
cuttings (IBA)
Plates 2b: C. adansonii stem
cuttings' bud sprouts are
impacted by coconut liquid
endosperm (CLE)
Plate 3:On Crateva adansonii stem
cuttings treated
with 100% Coconut Liquid
Endosperm, leaves have
developed (CLE)
Plate 4: A 300mg/L (IBA)-treated
stem cutting of the
Crateva adansonii plant
displaying several buds that
eventually turned into
leaves.
Bio-Research Vol.21 No.1
pp.2003-2018 (2023)
2011
Table 3: Effects of coconut
liquid endosperm (CLE) and indole butyric acid (IBA) on the percentage
of
sprouting buds that grow into
leaves on Crateva adansonii stem cuttings
Treatments
Percentage development of
sprouted bud into leaves (%)
IBA (mg/L)
100
73.25 ± 1.36
b
200
79.75 ± 0.85
a
300
82.75 ± 1.11
a
Control
59.75 ± 1.55
c
LSD (P<0.05)
3.84
CLE (%)
60
65.50 ± 0.65
c
80
72.50 ± 1.04
b
100
84.25 ± 2.17
a
Control
59.62 ± 1.07
d
LSD (P<0.05)
4.18
Values in the vertical column
that share the same alphabet (or alphabets) do not substantially
differ
from one another (P
≤0.05).
Table 4: Crateva adansonii stem
cuttings' percentage callus formation in response to indole butyric
acid (IBA) and coconut liquid
endosperm (CLE)
Treatments
Percentage callus (%)
IBA (mg/L)
100
46.50 ± 1.71
c
200
65.25 ± 1.25
b
300
81.75 ± 1.18
a
Control
0
LSD (P<0.05)
3.73
CLE (%)
60
28.00 ± 2.16
c
80
65.00 ± 1.22
b
100
81.25 ± 1.49
a
Control
0
LSD (P<0.05)
4.47
Values in the vertical column
that share the same alphabet(s) do not significantly differ
from one another (P
0.05).
Bio-Research
Vol.21 No.1 pp.2003-2018 (2023)
2012
Indole Butyric Acid (IBA) and
Coconut
Liquid Endosperm (CLE) impact on
root
growth of Crateva adansonii
Following studies on how the stem
cuttings
formed their roots, it was
discovered that the
stem cuttings developed two
different kinds of
roots: lateral roots and feeding
roots. The
results also showed that the stem
cuttings'
sides had few lateral roots and
only a few light
clustered feeding roots at the
cut-end.
Irrespective of the rooting
material and its
concentrations, the observations
were the
same on all rooted stem cuttings.
Plate 6
displays the outcome of the
rooted stem cutting
of Crateva adansonii treated with
300mg/L IBA
and displaying the main, lateral,
and feeder
roots.
Counting and measuring the length
of roots
on stem cuttings as influenced by
different
concentrations of IBA and
CLE
The number of roots, including
lateral and
feeder roots, per stem cutting
and the average
length of the five longest roots
per cutting are
determined. The results are
provided (Plate 6
and Table 5). In general, the
results revealed
diversity between the various
treatments,
including the different IBA and
CLE
concentration levels. Per stem
cutting, there
were anywhere between 15.25±0.48
to
25.50±1.32 roots. The stem
cuttings soaked in
300 mg/L IBA produced the most
roots
(25.50±1.32), followed by those
soaked in 200
mg/L IBA and 100 percent CLE
(19.25±0.48),
and the stem cuttings soaked in
60 percent CLE
(15.25±0.48) produced the fewest
roots. The
longest five roots on each stem
cutting ranged
in length from 8.13±0.05 cm to
8.60±0.04cm on
average. The results also showed
that stem
cuttings soaked in 200mg/L IBA
and 80% CLE
produced the longest roots
(8.60±0.04 cm).
Stem cuttings soaked in 300 mg/L
IBA
8.53±0.03cm followed, while stem
cuttings
soaked in 100 mg/L IBA
8.13±0.05cm had
shortest roots.
Comparatively, the findings
showed that stem
cuttings treated with different
concentrations of
IBA developed more roots on the
stem cuttings
than stem cuttings treated with
different
concentrations of CLE. Further
analysis of the
results revealed that CLE
concentrations of
100%, 80%, and 60%, which
produced
19.25±0.48, 16.00±0.41 and
15.25±0.48
correspondingly, had a greater
influence on the
development of roots than higher
IBA
concentrations (300mg/L and
200mg/L) did.
According to the findings, stem
cuttings treated
with IBA concentrations produced
longer roots
than stem cuttings soaked with
CLE.
Aerial growth parameters of
rooted stem
cuttings
Table 6 shows the results of
calculating the
mean values of some aerial growth
metrics of
the rooted stem cuttings,
including branch
count, branch length, and mean
leaf area.
On the stem cuttings, there
appeared
somewhere between 5.25±0.48 to
12.75±0.85
branches. The stem cuttings that
had been
treated in 300mg/L of IBA had the
most
branches on them, which was
followed by stem
cuttings soaked in 100 percent
CLE
(8.75±0.48). Stem cuttings bathed
in 100mg/L
of IBA had the least. The results
also
demonstrated that the branches'
lengths varied
from 3.75±0.48 cm to 8.75±0.48cm.
The
longest branches were formed by
stem cuttings
that had been immersed in 100
mg/L
(8±0.75cm) of IBA. Stem cuttings
bathed in
200mg/L IBA and 100% CLE followed
after that.
The seedlings' mean leaf area was
calculated,
and the results showed that it
ranged from
23.50±0.65 cm
2
to 26.75±0.48 cm
2
. The
broadest leaves, measuring
26.75±0.48, were
produced by stem cuttings
immersed in 100
mg/L IBA and 60 percent CLE. Stem
cuttings
immersed in 100 percent CLE
generated the
narrowest leaves
(23.50±0.65cm
2
), followed by
cuttings soaked in 300 mg/L IBA,
200 mg/L IBA,
and 80 percent CLE, which all had
leaves with
an area of 24.25±0.48cm
2
.
DISCUSSION
The results from the
meteorological data within
the period of the trial which
included: mean
monthly rainfall, humidity,
temperature from
May to October showed high
rainfall and
relative humidity incidence
alongside low
temperature. The results agree
with the earlier
reports of Araya et al. (2007)
and Sani et al.
(2016) who reported May to
October as the
period with best result in their
various trials.
Studies on rooting of stem
cuttings by (Sani et
al.,2016) were favoured by the
wet season,
characterized by high rainfall,
high humidity and
low
temperature.
Bio-Research Vol.21 No.1
pp.2003-2018 (2023)
2013
The first indication of life
renewal that might
lead to further development was
seen in the bud
breaking. Buds sprouted on all
stem cuttings,
including the control, regardless
of the rooting
chemicals used or their
concentrations
(untreated stem cuttings). On
stem cuttings,
the time between the first and
final bud
sprouting varied. This supports
past studies
that described the steps involved
in the
production of roots on stem
cuttings treated
with rooting agents, including
bud break, leaf
formation, callus formation, and
rooting (Gilani
et al., 2019). The development of
adventitious
roots is the primary process in
asexual
propagation. Adventitious root
growth is
influenced by both internal and
external
influences (Cibele et al., 2013).
The most
significant impact on internal
factors is
attributed to phyto-hormones,
with preference
to auxins but IBA has been
reported to have
greater ability to induce
adventitious roots
formation than indole acetic
acid-IAA (Ludwig-
Muller, 2000; Hakan and Kerim,
2013) and this
may be due to increased stability
of IBA. Similar
report was also upheld by
Ludwig-Muller (2000)
and Fattorini et al. (2017).
Therefore, indole-
butyric acid (IBA) has been
advocated as a
better hormone to encourage the
development
of roots in plant stem
cuttings.
According to the current study,
the great
susceptibility of the species to
bud sprouting
under any degree of concentration
of any
known rooting medium, maybe
related to the
species' genetic makeup, could be
the cause of
the bud breakage as observed on
all of the stem
cuttings, including the
control.
a b
Plates 5: The stem cuttings
treated with coconut liquid endosperm (CLE) in (a) and indole butyric
acid
(IBA) in (b) developed calluses
and roots.
Plate 6: Rooted stem cutting of
Crateva adansonii with main, lateral, and feeder roots after
treatment with 300mg/L indole
butyric acid IBA.
Bio-Research Vol.21 No.1
pp.2003-2018 (2023)
2014
Table 5: Effect of coconut liquid
endosperm (CLE) and indole butyric acid (IBA) on the quantity of
roots and average length per stem
cutting of Crateva adansonii
Treatments
Number of roots per
cutting
Mean length of five
longest
roots per stem cutting
(cm)
IBA (mg/L)
100
16.50 ± 0.65
c
8.13 ± 0.05
b
200
19.25 ± 0.48
b
8.60 ± 0.04
a
300
25.50 ± 1.32
a
8.53 ± 0.03
a
Control
0.00 ± 0.00
d
0
LSD (P<0.05)
2.38
0.01
CLE (%)
60
15.25 ± 0.48
b
8.43 ± 0.05
b
80
16.00 ± 0.41
b
8.60 ± 0.04
a
100
19.25 ± 0.48
a
8.40 ± 0.06
b
Control
0.00 ± 0.00
c
0
LSD (P<0.05)
1.24
0.131
Values in the vertical column
that share the same alphabet (or alphabets) do not substantially
differ
from one another (P
≤0.05).
Table 6: Some Crateva adansonii
aerial growth metrics after treatment with coconut liquid endosperm
(CLE) and indole butyric acid
(IBA)
Treatments
Number of branches
Length of branches
(cm)
Mean leaf area
(cm)
IBA (Mg/L)
100
4.75 ± 0.48
b
8.75 ± 0.48
a
26.75 ± 0.48
a
200
7.00 ± 0.71
b
8.00 ± 1.29
a
24.25 ± 0.48
b
300
12.75 ± 0.85
a
5.25 ± 0.75
b
23.50 ± 0.65
b
Control
0.00 ± 0.00
c
0.00 ± 0.00
c
0
LSD
1.85
2.42
1.28
CLE (%)
60
5.25 ± 0.48
b
4.00 ± 0.41
b
26.75 ± 0.48
a
80
5.75 ± 0.63
b
3.75 ± 0.48
b
24.25 ± 0.48
b
100
8.75 ± 0.48
a
6.25 ± 0.75
a
23.50 ± 0.65
b
Control
0.00 ± 0.00
c
0.00 ± 0.00
c
0
LSD
1.42
1.50
1.44
Values in the vertical column
that share the same alphabet (or alphabets) do not substantially
differ
from one another (P
≤0.05).
Bio-Research
Vol.21 No.1 pp.2003-2018 (2023)
2015
Since plant meristematic tissues
produce
hormones including cytokinin and
IAA, Azizi and
Sahebi (2015) suggested that
these hormones
may be to blame for the early bud
sprouting
observed on all of the stem
cuttings. The
discrepancy in the times that the
stem cuttings'
buds sprouted could be explained
by the
interaction between the hormones
that the plant
naturally produces and the
rooting agents (IBA
and CLE) that were administered.
A lack of
intrinsic hormone concentration
in the cutting
may have contributed to the
lengthier time it
took the control group to start
early bud
sprouting. This may be the real
cause of the
high rate of single and multiple
bud sprouting
seen in stem cuttings treated
with additional and
variable concentrations of CLE
and IBA.
There aren't many comprehensive
studies on
the growth of buds that emerged
from cuttings
to become photosynthesizing
leaves. In the
current investigation, it was
found that the
untreated stem cuttings had a
higher rate of
none sprouting into genuine
leaves than the
stem cuttings treated with low
quantities of IBA.
This may be because the hormones
supplied in
lesser concentrations were
insufficient to
encourage the production of
leaves. According
to Nzekwe (2002) and Sani et al.
(2016), not all
buds that emerged on stem
cuttings lasted
through leaf
development.
The findings demonstrated that
higher
concentrations of IBA (300 mg/L)
and Coconut
liquid endosperm (CLE) had a
greater influence
on the proportion of buds that
developed into
leaves, up to 83 mg/L for IBA and
84 mg/L for
CLE, respectively. The higher
concentrations of
rooting components that provided
the stem
cuttings with additional rooting
substance for
subsequent developmental
activities appear to
have had an impact on the higher
percentages
of bud development into leaves.
The 83 percent
successful bud development into
leaves on
300mg/L IBA as opposed to 84
percent
impacted by 100 percent CLE
reveals that CLE
performs better than the former
and points to
the need for future studies to
use IBA
concentrations above 300mg/L for
better
results. Low percentage of
untreated stem
cuttings' buds developing into
leaves could be a
result of depletion of the
naturally produced
auxin in the stem cuttings, thus
the use of
untreated stem cuttings of
Crateva adansonii in
the vegetative propagation of the
species may
not be hoped upon for the routine
generation of
the species seedlings. The
transformation of
buds into leaves suggested that
additional
nutrients were being produced
by
photosynthesis in the leaves.
According to
Nzekwe (2006), a plant's capacity
to absorb
nutrients through photosynthesis
increases in direct
proportion to the number of
healthy leaves it
produces. This rise is mostly
attributable to auxins,
which promote photosynthesis,
participate in the
light reaction, and activate the
enzyme rubulose-
bisphosphate carboxylase,
increasing leaf nutrition
(Al-hasnawi , 2012). This is
consistent with research
done by Amrut and Rajput (2013)
on fenugreek and
Ullah et al. (2013) on marigold
(Tagetes erecta L.).
The observations made on the stem
cuttings
demonstrated that callus
production was not implied
by the appearance of bud rupture
and eventual leaf
development seen in all of the
stem cuttings. These
results agree with those of Puffy
et al (2008). The
development of two
morphologically different types
of calluses on the stem cutting
of the same plant
species has been documented by a
number of
authors (Nzekwe, 2006; Puffy et
al.,2008; Sani et
al., 2016). On stem cuttings
treated with rooting
agents prior to rooting, they
also observed the
development of two
physiologically distinct types of
calluses. They classified the
first callus type as
"heavy calluses,"
characterized by heavy swelling of
the stem cuttings and basal
region, associated with
vertical swelling. According to
Norhayati et al.
(2013), strong callus
proliferations were a reliable
indicator of how quickly stem
cuttings will root. The
second callus type was also
referred to as "light
calluses" by the authors and
was capable of
producing seedlings (rooted stem
cuttings) with just
lateral and feeder roots and no
tap root system.
The findings of the current
investigation
demonstrated that stem cuttings
associated with
seedling formation with just
lateral and feeder roots,
without a tap root system, were
only mildly callused,
regardless of the concentration
of rooting agent.
This is also consistent with what
Puffy et al.(2008)
observed. Despite light calluses
noticed on stem
cuttings, irrespective of the
concentrations of the
rooting substance, cuttings
treated with higher
concentrations of IBA (300mg/L,
200mg/L) and CLE
(100%, 80%) had higher
percentages of callusing
than those treated with lower
concentrations of IBA
(100mg/L) and CLE (60%). The
production of light
calluses observed in this study
could be an
indication that further
investigation involving the use
of higher concentrations of IBA
or other rooting
substance should be encouraged to
find out if there
could be changes in aerial and
basal growth
parameters. The higher
concentrations of IBA (300
mg/L) and CLE (100%) is believed
to be responsible
for the highest percentage
rooting observed on the
stem cuttings up to 80% out of
all the already
callused stem cuttings leaving
the control with no
root
development.
Bio-Research
Vol.21 No.1 pp.2003-2018 (2023)
2016
The inability of the control to
develop root could
possibly have been because of the
insufficiency
of the available hormone to have
initiated root
formation. The breaking of
hydrogen bonds
between cellulose microfibrils by
proteins from
IBA is what causes the trend in
root length,
encouraging cell wall thinning
and eventual cell
enlargement (Kumar et al., 2015;
Qu Yang et
al., 2015). The rate of cambium
dedifferentiation
is raised, hydrolytic activity is
accelerated, and
callus production is boosted at
optimal
exogenous IBA, all of which
contribute to better
root length (Gilani et al.,
2019). The action of
the cytokinins and auxin in
coconut water may
be to blame for this. Thus, it is
clear that the
presence of cytokinins and IBA in
the coconut
water encouraged the growth of
large numbers
of undifferentiated cells
(callus). After being
exposed to certain hormones, such
as auxins
(found in coconut water), which
formed roots,
the callus cells are then
encouraged to grow
roots. The influence of growth
regulators IBA on
the translocation of metabolites
and metabolism
of carbohydrates may also be
responsible for
the increase in root length. When
compared to
the other auxines, Hakan and
Kerim (2013) saw
that IBA produced a larger output
of roots in
Melissa officinalis. The outcome
is in line with
Okunlola (2016) investigations,
which found that
rooting hormones applied to
Bougainvillea
spectabilis wood cuttings had a
substantial
impact on the length of roots
compared to the
control. These findings concur
with those of
Oluwagbenga (2016) who discovered
that
coconut water treated Parkia
biglobosa plant
had more roots per plant than the
control. Given
the reduced time needed for
callus formation
and the increased cambium
dedifferentiation,
Singh et al. (2014) noted that
there are more
roots at the optimal IBA
concentration. These
numerous cells will differentiate
into root cells.
The pace of disappearance of
amyloplasts is
accelerated by optimal IBA.
During rooting,
amyloplast levels naturally
decrease (Singh et
al., 2014). Our results show that
at optimal
indole-3-butyric acid, amyloplast
decline can be
improved and cambium activities
are stimulated,
which will mobilize stored food
supplies to the
root initiation site and
encourage the production
of many roots (Gilani et al.,
2019).
From the study’s findings which
revealed that
rooted stem cuttings did not
produce taproot
system implied that either the
concentrations
of the chemical substances were
not sufficient
to have an impact on seedling
production that
resembles the parent stock, or
the seedlings
require more growth in the
nursery. Despite
producing seedlings that had no
taproot
system, the understudy species
can be multiplied
using stem cuttings and the
vegetative propagation of
the species can immensely
contribute to the
production of large quantity of
seedlings which can
help in the future exploitation
of the species medicinal
potentials. Such seedlings would
be uniform and from
that resources can be exploited
from the species as
noted by Lodama and Robbertse
(2016). Based on the
high percentage callusing of the
cuttings treated with
300mg/L IBA, which had no
taproots, there is need to
evaluate the effects of other
rooting substances at
higher concentrations on
production of species
seedlings that resembles the
parent stock. The
finding is in conformation with
the report of Sani et al.
(2016) on the investigation on
Moringa oliefera that
showed that IBA and IAA
respectively influenced the
development of aerial and sub
aerial parameters of
the plant.The same concentrations
of the treatments
in the current study produced
larger values of plant
aerial growth characteristics
like number of leaves,
length of branches, mean leaf
area, and sub-aerial
growth parameters like number of
roots and root
length than their lower
concentrations did. The
development of higher values of
the aerial and sub-
aerial growth parameters appears
related to the early
periods of bud breakage and leaf
development.
CONCLUSION AND
RECOMMENDATIONS
This study’s findings allow us to
draw the conclusion
that Crateva adansonii can be
multiplied by treating
the stem cuttings with the
appropriate concentration
of IBA preferably above 300 mg/L.
The high
percentage of rooting response of
cuttings treated
with 300mg/L IBA and 100% CLE
concentrations can
be dependable for a high volume
of seedling
production required for the
species genetic resource
conservation in large proportion
needed for future
biotechnological exploitation for
its medicinal
bioactive potentials. However,
further studies could
be encouraged especially with the
use of other
rooting substances. Low
percentage development of
buds on low concentration treated
stem cuttings of
the respective rooting substances
implied that the
concentrations of the rooting
chemicals were not
adequate for obtaining good
responses by the stem
cuttings, hence higher
concentrations are
recommended for further studies.
Based on the high
percentage callusing of the
cuttings treated with 300
mg/L IBA, which had no tap roots,
there is need to
evaluate other rooting substances
at higher
concentrations on production of
species seedlings
that resemble the parent stock.
The production of
light calluses observed in this
study implied that
further studies involving the use
of higher
concentrations of IBA or other
rooting substance can
be investigated to find out if
there could be changes in
aerial and basal growth
parameters as well as heavy
callus
formation.
Bio-Research
Vol.21 No.1 pp.2003-2018 (2023)
2017
Conflict of interest
The authors have no conflict of
interest to
declare
Author Contribution
Study design – NMC, ON, NNO, OVO,
OAA,
EOC, data collection – NMC, ON,
NNO, data
analysis – NMC, OVO, manuscript
writing –
NMC, ON, EOC, OVO, OAA, critical
revision –
NMC, ON, NNO, EOC, study
supervision –
NMC, ON, NNO, OVO
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