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Rooting Ability of Cocoa (Theobroma cacao L.) Stem Cuttings: Effect of

Genotype, Cutting Type, Hormone Concentration and Their Interactions

Article in Asian Journal of Agricultural and Horticultural Research · March 2018

DOI: 10.9734/AJAHR/2018/40168

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 Asian Journal of Agricultural and Horticultural Research

1(2): 1-10, 2018; Article no.AJAHR.40168

Rooting Ability of Cocoa (Theobroma cacao L.) Stem

Cuttings: Effect of Genotype, Cutting Type,
Hormone Concentration and Their Interactions
K. M. D. Kamga1*, D. T. Tchatchoua1, R. G. Caspa2, G. Yombo A. Bessa3
and L. J. Baleba3
1
Department of Agriculture, Animal Husbandry and Derived Products, National Advanced School of
Engineering, University of Maroua, B.P.46, Maroua, Cameroon.
2
Institute of Agricultural Research for Development (IRAD), Forest and Wood Program,
P.O.Box 2123, Yaounde, Cameroon.
3
Institute of Agricultural Research for Development (IRAD), Nkoemvone, P.O.Box 65, Ebolowa,
South Region, Cameroon.

Authors’ contributions

This work was carried out in collaboration between all authors. Author KMDK designed the
experiment, carried out the research and wrote the first draft of the manuscript. Author DTT initiated
the concept, designed the experiment, performed the statistical analysis and wrote the article. Author
RGC wrote the article with literature searches. Authors GYAB and LJB designed the experiment,
wrote the protocol and managed the analysis of the study. All authors read and approved the final
manuscript.

Article Information

DOI: 10.9734/AJAHR/2018/40168
Editor(s):
(1) Fatemeh Nejatzadeh-Barandozi, Department of Horticulture, Faculty of Agriculture, Azad Islamiz University of Khoy, West
Azarbayjan, Khoy, Iran.
Reviewers:
(1) Bado Souleymane, University of Natural Resources and Life Sciences, Austria.
(2) R. Mahalakshmi, India.
Complete Peer review History: http://www.sciencedomain.org/review-history/23865

Received 11th January 2018

th
Accepted 20 March 2018
Short Research Article th
Published 28 March 2018

ABSTRACT
Aims: This study aimed to evaluate the rooting ability of stem cuttings of eight-year-old cocoa
th
hybrids disseminated in the 5 agroecological zone of Cameroon.
Study Design: The experiment was a 6 x 2 x 3 randomised complete block design with three
replications.
Place and Duration of Study: The study was conducted at the multipurpose agricultural research
station Nkoemvone – Ebolowa from February to September 2017.
_____________________________________________________________________________________________________

*Corresponding author: Email: kadimitri02@yahoo.fr;

 Kamga et al.; AJAHR, 1(2): 1-10, 2018; Article no.AJAHR.40168

Methodology: The studied factors were six genotypes (G1 = IMC67 x SNK 64, G2 = SNK 620 x
MA 12, G3 = T79/501 x SNK 109, G4 = T79/501 x SNK 64, G5 = SNK 413 x UPA 143 and G6 =
UPA 143 x SNK 64), two Cutting types (C1 = Orthotropic and C2 = Plagiotropic) and three rooting
hormone concentrations (H1 = 2 tablets per liter of water, H2 = ½ tablet per liter of water and H3 =
1 tablet per liter of water). Rooting ability was evaluated by assessing percentage of rooted
cuttings, number of root per rooted cutting and the length of the longest root per rooted cutting.
Results: Analysis of variance results indicated that genotype, hormone concentration and most of
their interactions were highly significant while cutting type was not significant on the measured
traits. The best rooting was obtained with genotype G4 for all the measured parameters. It was
shown in this experiment that genotype G4 could be propagated using Rhizopon hormone, with a
concentration of ½ tablet per litre of water and plagiotropic cutting type. This is also noticed in their
interaction terms, where genotype G4/Hormone concentration 2 and 3 were the best combinations
for all parameters.
Conclusion: Significant differences among genotypes are an indication that different genotypes
may require different conditions for their propagation. As such investigations into the requirement
for the propagation of the other genotypes should be considered in the future.

Keywords: Theobroma cacao; vegetative propagation; hybrid; cutting type; rooting; hormone
concentration; 5th agroecological zone.

1. INTRODUCTION banks in which clones from diverse origins and

characteristics are planted. These living
Cocoa (Theobroma cacao L.) is a neotropical genebanks are found primarily at the Institute
perennial plant belonging to the family of Agricultural Research and Development
Malvaceae [1] which develops naturally around (IRAD) station of Nkoemvone in the South
the equator between latitudes 20o North and Region and maintained by vegetation
South [2]. The principal objective of cocoa propagation. Despite the fact that Cameroon
cultivation in all countries is to obtain cocoa makes a considerable contribution to
beans which constitute a significant raw material global cocoa production, mean yields (300 kg)
for food, pharmaceutical and cosmetic industries per hectare remain low [5]. This corresponds
[3,4]. Cocoa is cultivated in America, Asia and to ten times less its standard potential
Africa where it contributes significantly to their when cocoa is cultivated under optimum
economies, especially those of West African conditions [12]. Factors responsible for the poor
countries [5]. The latter sub-region contributes yields include:- cocoa brown pod rot disease
about 70% of global production of cocoa beans caused by the fungus (Phytophthora megakarya)
[6,7]. and mites,Sahlbergella singularis and Distantiella
theobromae [13,14] ageing of cocoa plantations
Cameroon is the fifth world producer of cocoa and farmers as well as bad agricultural practices
beans after Ivory Coast, Ghana, Indonesia, and [15-18] have also contributed to reduced
Nigeria [7]. The cocoa chain represents about production. According to Mahob [19], lack of
3% of Gross Domestic Product, 6% of the improved planting material contributes to low
primary GDP and about 30% of the agricultural cocoa production.
sub-sector destined for exportation and
transformation [8]. Cocoa is cultivated on an area The future of cocoa in Cameroon as inscribed
of about 400.000 to 600.000 hectares spread in the cocoa and coffee development plan by
within the humid forest zone [9]. Annual 2020 can only be attained through increase
production during the 2014/2015 and 2015/2016 of seed banks, replanting of old plantations,
seasons were respectively, 232.530 and 269.495 establishment of new plantations, which
tons of marketable cocoa beans [10,11]. The necessitate an increase in demand for
monetary value of exported cocoa beans in improved planting materials. Many structures and
Cameroon is estimated at 200 billion francs CFA projects have been put in place by the
and is a source of sustenance for about 2 million government such as project for the selection and
people distributed among about 400.000 diffusion of improved cocoa and coffee planting
producer families [8]. material (SDMVCC), Fund for the development
of cocoa and coffee chains (FODECC) and
The maintenance of cocoa genetic diversity in Cocoa development company (SODECAO).
Cameroon is done by the establishment of gene They are responsible for the production and

2
 Kamga et al.; AJAHR, 1(2): 1-10, 2018; Article no.AJAHR.40168

dissemination of disease resistant and improved 2. MATERIALS AND METHODS

planting material such as hybrids. This is
achieved in IRAD through controlled manual 2.1 Experimental Site and Establishment
pollination and the establishment of multi-clonal of Cuttings
seed orchards from improved local clones
belonging to different genetic groups [20]. The The experimental site and propagation procedure
first flowering of such hybrids occurs 5 to 7 have been previously reported by [24]. However,
years after planting [14]. Multi-clonal seed the genotypes used in this experiment consisted
orchards produce hybrids in a heterogenous of six hybrids (genotypes) G1 = IMC67 x SNK
manner with an uncertainty of individuals' 64, G2 = SNK 620 x MA 12, G3 = T79/501 x
identities since pollination takes place naturally. SNK 109, G4 = T79/501 x SNK 64, G5 = SNK
Studies in multi-clonal seed orchards show 413 x UPA 143 and G6 = UPA 143 x SNK 64 of
that about 97% of cocoa beans obtained results age 8 years old. These genotypes were chosen
from self-pollination [21,22]. According to the based on their good performance concerning
latter authors, manual pollination requires yield, growth, resistance to pests and diseases
qualified personnel and sophisticated equipment and other morphological characteristics (Fig. 1).
but gives a low rate of success (7 to 29.2 %).
The same difficulties encountered in the The trial was a 6 x 2 x 3 randomized complete
development of hybrids are sometimes block design with three replications. Cuttings
responsible for the high prices charged by were collected early in the morning (before 7 am)
producers during sale [23]. There is a necessity from afore mentioned six cocoa hybrids, from C1
for a method of rapid production of these hybrids = orthotropic (upright stems) and C2 =
from improved cocoa planting material. The plagiotropic (side) branches. Cuttings had an
objective of this study was to evaluate the rooting average of four leaves and were about 30 cm
ability of cuttings of selected cocoa hybrids used long. These were quickly dipped into three
in the humid forest zone of Cameroon. different concentrations of indolebutyric acid
Specifically, the study aimed at assessing the (IBA) (H1: two tablets per litre of water; H2: ½
effect of genotype, hormone concentration, tablet per litre of water and H3: 1 tablet per litre
cutting type and their interactions on rooting of of water) before setting in, decomposed saw dust
stem cuttings. disinfected three days before. Alkatene plastic

G1 = IMC67 x SNK 64 G2 = SNK 620 x MA 12 G3 = T79/501 x SNK 109

G4 = T79/501 x SNK 64 G5 = SNK 413 x UPA 143 G6 = UPA 143 x SNK 64

Fig. 1. Observed characteristics of the six genotypes

3
 Kamga et al.; AJAHR, 1(2): 1-10, 2018; Article no.AJAHR.40168

pots (24 x 14 cm) containing cuttings were then 3. RESULTS

placed in propagators containing trays filled with
disinfected wood chips to provide stability. The 3.1 Effect of Genotype on the
experimental unit consisted of 20 cuttings each Percentage of Rooted Cuttings,
giving a total of 2160 cuttings (20 cuttings x 6
Number of Roots per Rooted Cutting
genotypes x 2 cutting types x 3 hormone
concentrations x 3 replications). The trial was and Length of the Longest Root
watered daily in the morning and evening
whereas fallen and dead leaves were removed The measured parameters varied from 45 to
as reported by [25]. 90% for the percentage of rooted cuttings, 1 to
16 for the number of roots per rooted cutting
2.2 Data Collection and Analyses and 3 to 33.6 cm for the length of the longest
root among the genotypes. Results of analysis
Percentage of rooted cutting (rate of survival) of variance show that genotype has significant
was determined by the proportion of life cuttings effect on the percentage of rooted cuttings,
at the end (after about 3 months) of the the number of roots and the length of the
experiment to the number of cuttings set. The longest root at 5% level with P < .000. The
number of roots per rooted cutting was counted DMRT test for mean comparison shows that
to determine root production, whereas length of genotype T79/501 x SNK 64 had highest values
the longest root per rooted cutting was measured in all the measured parameters with 85% ± 5.47
using a ruler. These data were entered into for the percentage of rooted cuttings, 10.05 ±
Microsoft Excel 2013 and analysed using SPSS 2.36 for the number of roots per rooted
IBM version 20. Normality test and homogeneity cutting and 13.84 cm ± 4.77 for the length of the
of variance test were conducted to confirm the longest roots and was significantly different
validity of tests of analysis of variance. Means from all the other genotypes used, whereas
were separated using Duncan Multiple range test SNK413 X UPA 143 showed the least
(DMRT). The univariate analysis of variance percentage of rooted cuttings, number of roots
(ANOVA) with principal factors: genotypes, per cutting and length of the longest root with
hormone concentrations, cutting types and 50.00 ± 6.32%, 3.28 ± 0.87 and 4.76 cm ± 1.10
interactions: genotype x hormone concentration respectively (Table 1).
and genotype x cutting type to test the
significance between different parameters at 5 % 3.2 Effect of Hormone Concentration on
level using the GLM type III model as follow: the Percentage of Rooted Cuttings,
Number of Roots per Rooted Cutting
Yijkl = µ + Gi + Hj + Tk + Rl + GiHj + GiTk + εijkl and Length of the Longest Root
Y = Observation for the ijklth individual, µ = Descriptive statistics of the mean data obtained
overall mean of the experiment, Gi = the random among the hormone concentrations show that it
th
effect of the i genotype, Hj = the fixed effect of varies from 61.25 to 73.61% for the percentage
th
the j hormone concentration, Tk = the fixed of rooted cuttings, 5.17 to 6.53 for the number of
th
effect of the k cutting type, Rl = the random root and 7.76 to 9.19 cm for the length of the
effect of lth replication, GiHj = interaction effect of longest root. Result indicated that the percentage
th th
i genotype x j hormone concentration, GiTk = of rooted cuttings was best (73.61% ± 12.30) in
interaction effect of ith genotype x kth cutting type, hormone concentration H2 (½ tablet per litre of
εijkl = the sampling error water) which was not significantly different from

Table 1. Mean values of percentage of rooted cuttings, number of root per rooted cuttings and
the length of the longest root per cutting among genotypes

Genotype Number of Percentage of Number of root Length of the

cuttings rooted cuttings per rooted cutting longest root
e d e
G1 (IMC67 x SNK 64) 360 60.00 ± 8.94 4.30 ± 1.47 5.57 ± 1.50
G2 (SNK 620x MA 12) 360 65.00d ± 10.95 4.88c ± 1.38 6.61d ± 1.48
G3 (T79/501x SNK 109) 360 80.00b ± 10.48 6.97b ± 2.14 11.71b ± 2.29
a a a
G4 (T79/501x SNK 64) 360 85.00 ± 5.47 10.05 ± 2.36 13.84 ± 4.77
G5 (SNK 413x UPA 143) 360 50.00f ± 6.32 e
3.28 ± 0.87 f
4.76 ± 1.10
c b c
G6 (UPA 143x SNK 64) 360 75.00 ± 10.85 6.56 ± 1.40 9.79 ± 1.40
P - value P < .000 P < .000 P < .000

4
 Kamga et al.; AJAHR, 1(2): 1-10, 2018; Article no.AJAHR.40168

hormone concentration H3 (1 tablet per litre of cutting and the length of the longest root with
water). Results of ANOVA revealed that both orthotropic and plagiotrophic cuttings each
hormone concentration had a highly significant producing 6 roots per cutting and 8.1 cm for the
effect on number of roots with P = 0.000. There length of the longest root at 5%, with P = 0.87
was no significant difference in root number (Table 3).
between ½ and 1 hormone tablet per litre of
water, which induced the greatest number of 3.4 Interaction Effect between
roots with mean values of 6.53 ± 2.83 and 6.33 ± Genotype/Hormone Concentration on
2.86 respectively. The least number of roots was the Percentage of Rooted Cuttings,
produced by hormone concentration H1 with 2 the Number of Roots and Length of
tablets per litre of water (Table 2).
the Longest Root
ANOVA results reveal that hormone
concentration had a significant effect on length of Significant interaction effect was found in
the longest root at the level of 5 % with P = 0.000 genotype/hormone concentration on the number
(Table 2). It was observed that the longest root of rooted cuttings (P = 0.012) and the length of
length was induced by ½ and 1 hormone tablet the longest root (P = 0.00). There was no
per litre of water, which showed no significant significant effect on the percentage of rooted
difference at the 5% level, with mean root lengths cutting and the results could not be reported
of 9.19 ± 4.17cm and 9.18 ± 4.41 cm. further. The highest mean number of root per
rooted cutting was obtained from the interactions
3.3 Effect of Cutting Type on the between genotype 4 and hormone
Percentage of Rooted Cuttings, concentrations 2 and 3, which showed mean root
Number of Root and Length of the per cutting as 10.7 and 10.5 respectively (Fig. 2,
Longest Root 3). This was followed by genotype 4/ two tablets
per litre of water (8.8 mean roots). For the mean
According to the ANOVA results, cutting type had number of roots genotype, 4 was highest in all
significant influence on the percentage of rooted the concentration applied. Genotype 5/hormone
cutting at 5 %, with P = 0.025 with plagiotrophic concentration 1 had the lowest mean number of
cuttings showing the highest (70.37 ± 15.55) roots with 2.9. It was shown that hormone
percentage of rooted cuttings. Cutting type had concentration 1 recorded the lowest mean
no influence on the number of roots per rooted number of roots for all the genotypes.

Table 2. Mean values of percentage of rooted cuttings, number of root per rooted cuttings and
the length of the longest root per cutting among hormone concentrations

Hormone Number of percentage (%) of Number of Length of the

concentration cuttings rooted cuttings roots per longest root
rooted cutting (cm)
b b b
2 tablets per liter 720 61.25 ± 12.67 5.17 ± 2.39 7.76 ± 3.27
of water
½ tablet per liter 720 73.61a ± 12.30 6.53a ± 2.83 9.19a ± 4.17
of water
a a a
1 tablet per liter of 720 72.63 ± 15.20 6.33 ± 2.86 9.18 ± 4.41
water
P - value P <.000 P <.000 P <.000

Table 3. Mean values of percentage of rooted cuttings, number of root per rooted cuttings and
the length of the longest root per cutting among cutting types

Cutting type Number of percentage of Number of root per Length of the

cuttings rooted cuttings rooted cutting longest root
Orthotropic 1080 67.963 b ± 14.16 6.006 a ± 2.73 8.731 a ± 4.06
Plagiotropic 1080 70.370 a ± 15.55 6.020 a ± 2.8 8.700 a ± 4.01
P - value P = .025 P= .873 P = .817

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 Kamga et al.; AJAHR, 1(2): 1-10, 2018; Article no.AJAHR.40168

The longest roots were induced by interactions and Lass [3], reported that differences in cocoa
between genotype 4/concentration 2 and 3, with genotypes lead to differences in their rooting
mean root lengths of 14.6 and 14.8 cm abilities. The fact of having obtained the best
respectively (Fig. 4, 5). Other interesting results rooting parameters with hybrid T79 / 501 x
came from interactions between genotype SNK64, T79 / 501 x SNK 109, UPA 143 x SNK
3/concentration 3 (12.4 cm), genotype 64, could be explained by the fact that in these
3/concentration 2 (12.2 cm), genotype hybridizations the African clones are involved
4/concentration 1(12cm). On their part, (SNK : Cameroonian clone, and T79 / 501:
interactions between genotype 6/concentrations Ghanaian clone), thus corroborating with the
3, 2 and genotype 3/concentration 1 each observations of [26], who reported that
produced longest roots of about 10 cm. Amazonian high clones such as UPA 143, and
those belonging to the Trinitario group such as
4. DISCUSSION T79 / 501 and the SNK, are more efficient during
cuttings, and therefore root better. For the effect
The results obtained on the genetic potential give of hormone concentration, it was also observed
us information on the effectiveness of the that the higher hormone concentration had a
differences in the rooting of the cuttings from the negative impact on the percentage of rooted
difference in the genetic potential of the hybrids. cuttings whereby hormone concentration H1 (2
Current literature on cocoa propagation does not tablet per litre of water) had the lowest (61.25 ±
provide information on the rooting of cuttings 12.67) mean percentage of rooted cuttings. In
taken from hybrids, as cuttings are often the test of the effect of concentration on the
collected mostly from clones. Similar results were rooting of cuttings, the results showed that the
obtained by Hall [26]. Toxopeus [27], and Wood concentration of the rhizogenic hormone solution

Fig. 2. An interaction effect between genotype/Hormone concentrations on number of roots

Fig. 3. Age of rooted cuttings, a) Genotype 3/Two Rhizopon tablets per litre of water b)
Genotype 3/½ Rhizopon tablet per litre of water c) Genotype 4/1Rhizopon tablet per litre of
water

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 Kamga et al.; AJAHR, 1(2): 1-10, 2018; Article no.AJAHR.40168

Fig. 4. An interaction effect between genotype/Hormone concentration on length of the longest

roots

a b c

Fig. 5. Length of the longest root a) Genotype 4/ hormone concentration 2 b) Genotype 4/

hormone concentration 3 c) Genotype 4/ hormone concentration 1

has indeed a consequence on the rooting; these economically important, but also reduce plant
results corroborate those from the [28, 27]. Wood production failures that sometimes occur when
and Lass [3] and Essola et al. [24] experiments, the hormone is missing or being not easily
which reported that hormones are one of the accessible.
important factors influencing the rooting of
cuttings; having obtained better rooting results at The observation of the non-significance of the
the lower concentrations of 1 and 0.5 tablets per rooting differences of cuttings according to
liter rather than the highest concentration of 2 whether they are orthotropic or plagiotropic, is in
tablets per liter, may be due to the fact that the line with the results obtained by Murray [30] cited
hormone used is based on auxin, slowing the by Lockwood [31], who reported that the root
activity of stimulation of rhizogenesis, due to an system of cocoa plants grown from rooted
excessive concentration of auxin, as underlined plagiotropic cuttings is similar to that of seedlings
by Charvet-Candela [29]. However, the non- grown from seeds. Several authors [32, 33, 8],
significance of the rooting results recorded for report that the plants resulting from the rooting of
hormone solution concentrations of ½ tablet per plagiotropic cuttings adopt a bulky canopy in
liter and 1 tablet per liter both gave better results, culture and have a low resistance to lodging and
it would be sensible and recommendable for water stress, this being explained in that these
users of Rhizopon, opting for the lowest plants do not develop a taproot, unlike plants
concentration, ½ tablet, which could not only be resulting from the cutting of orthotropic branches;

7
 Kamga et al.; AJAHR, 1(2): 1-10, 2018; Article no.AJAHR.40168

in the case of this study, these pivotal roots have for all the measured parameters, while no
not been observed on orthotropic cuttings that significant effect between genotype and cutting
have been experimented; this could be explained type. With the increasing demand of planting
by the stage of root development, perhaps not materials in this zone of Cameroon it is
advanced enough to be able to distinguish recommended on the basis of this work carried
between lateral roots and pivoting roots. These out to evaluate the cutting ability of cocoa hybrids
same authors as well as Lee [34] emphasized to clone the cocoa hybrids by cuttings. This will
that this lack of taproot is a disadvantage for not only reduce the time it takes for stock plants
cultivation, especially during periods when to flower but also reduce the problems with
rainfall is not sufficient; However, Boulay [35] sophisticated equipment required by manual
considers that this lack of root actually affects pollination.
little the mineral nutrition of the plant, because
the cocoa tree is a humicole species, which have ACKNOWLEDGEMENTS
nearly 85% of its roots grouped in the surface
horizon in the first 25 cm approximately. The first author is grateful to IRAD Nkoemvone
Interaction effects have never been studied in for permitting him assessed their structure used
propagation techniques in cocoa and comparison their facilities and work staff during the internship
can only be done using some tropical forest and period.
fruit trees. Significant interaction effect between
genotype and hormone (NAA) in rooting ability of COMPETING INTERESTS
P. santalinoides was reported by Dembele et al.
[36]. Authors have declared that no competing
interests exist.
5. CONCLUSION
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