Improving propagation methods of Ricinodendron heudelotti Baill. from cuttings

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Improving propagation methods of Ricinodendron heudelotti Baill.from cuttings

Nehemie Donfagsiteli Tchinda a,⁎, Hubert Jean Claude Mbita Messi a, Fotso b, Germo Nzweundji a,Nole Tsabang a, Bernard Dongmo a, Doungous Oumar c, Protus Arrey Tarkang a,Aaron Caver a, Denis Omokolo Ndoumou d

a Medicinal Plants and Traditional Medicine Research Centre, Institute of Medical Research and Medicinal Plants Studies (IMPM), P.O. Box 6163, Yaounde, Cameroonb Laboratory of General Biology, Higher Teacher's Training College, P.O. Box 39, Bambili, Cameroonc Jay P. J. Biotechnology Laboratory, Institute of Agricultural Research for Development, P.O. Box 25, Buea, Ekona, Cameroond Laboratory of Biochemical and Plant Physiology, Department of Biological Sciences, Higher Teacher's Training College, P.O. Box 42, Yaounde, Cameroon

a b s t r a c ta r t i c l e i n f o

Article history:Received 14 January 2012Received in revised form 22 February 2013Accepted 30 April 2013Available online xxxx

Edited by NJ Taylor

Keywords:Cutting typeGrowth regulators and honeyPropagationRicinodendron heudelotii

Ricinodendron heudelotii is a valuable multipurpose tree from Africa that is lacking an efficient and inexpen-sive vegetative propagation method. To improve multiplication, a series of nursery experiments wereconducted to assess the effect of propagation media, plant growth regulators (NAA and IBA), accession sourceharvest timing, cutting type, and pre-treatment with honey. Maximum survival and rooting percentages of90.2 and 93.7% respectively were achieved in all experiments. The maximum number of leaves and rootswere respectively, 7.4 and 8.7; maximum root length was 15.6 cm. Fine sand was superior to sawdust orfine sand:sawdust mixture. IBA, at 100 μg/l was more effective than NAA for rooting and growth. Principalcomponent analysis showed that, independent of the accession source, cuttings harvested from April toMay were more suitable for propagation, giving the highest rates of rooting and growth. In contrast, variableresults were achieved with apical cuttings (highest rooting), basal single-node leafy cuttings (highest num-ber of leaves and length of root) and basal two-node leafy cuttings (highest number of roots). Cuttings weremore successful when they were soaked in honey for 60 min. After acclimatization, maximum survival rate ofplants was 67.6%. This improved protocol can be incorporated into agro forestry system in which propagationof R. heudelotii is a component.

© 2013 SAAB. Published by Elsevier B.V. All rights reserved.

1. Introduction

Ricinodendron heudelotii Baill. (Euphorbiaceae), is a tree distribut-ed from Guinea to Angola and in East Africa; heights range from 12 to40 m. The species grows in secondary forest between 130 and1030 m above sea level (Vivien and Faure, 1996). It provides food,medicines and other various products, including constructionand building materials. The kernels are used as a flavoring agent,and are also a good source of oil used to make soap and varnish(Plenderleith, 2004; Tchiegang et al., 2005). The wood is used formaking household implements, drums and carvings (Plenderleith,1997); the bark is used to treat elephantiasis (Plenderleith, 2004).

Natural regeneration rates of R. heudelotii is low and is exasperat-ed by the negative impact of over exploitation has on natural popula-tion levels. This plant has therefore been classified as one of the topten species to be domesticated in Central and West Africa by the In-ternational Centre of Research in Agro Forestry (Franzel et al.,1996). In the case of R. heudelotii, the Institute of Medical Research

and Medicinal Plants Studies in Cameroon (IMPM) worked withfarmers to identify priorities and traits preferences for the develop-ment of vegetative propagation techniques.

Efficient propagation techniques will greatly facilitate the domes-tication and the improvement of the species. In previous studies onregeneration of R. heudelotii by cuttings, Shiembo et al. (1997a) usedseedlings or coppice shoots as stockmaterial in a non-mist propagationsystem to study the effect of indol-3-butyric acid (IBA), leaf area andplanting media on rooting. This low-technology system is designed foruse in rural areas and has successfully been used for the propagationof a large number of other trees species such as Irvingia gabonensis(Shiembo et al., 1997b), Prunus africana (Tchoundjeu et al., 2002a,2002b) and Vitex madiensis (Mapongmetsem, 2006). Mapongmetsemet al. (1999) used sprouts from stump of adult trees of R. heudelotii toinvestigate the effect of wood type and the cutting orientation onrooting. In both of these studies, rooting percentage was low. However,very little further research has been done on vegetative propagation ofR. heudelotii. In an attempt to further define appropriatematerial for usein the development of an efficient regeneration system, the effect of thephysiological status and origin of the cutting material on rooting andsubsequent plant regeneration of R. heudelotii was investigated, with

South African Journal of Botany 88 (2013) 3–9

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the ultimate objective of determining themost successful treatment formass production of clonal planting stock. The effects of propagationmedia, auxin concentration and pre-treatment with honey were alsoassessed.

2. Materials and methods

2.1. Study site and stock plant production

This study was conducted from September 2008 to December2009 in a propagation unit established in the nursery at IMPM atYaounde in the Centre region of Cameroon, located at latitude 3° 51′N and longitude 11° 25′ E., approximately 813 m above sea level.Average rainfall is 1692 mm and is bimodally distributed (Segalen,1967).

Three sites were chosen for seed collection: Yaounde (site de-scribed above) and Mbalmayo (latitude 3° 31′ N, longitude 11° 30′E, rainfall 1650 mm, altitude 600 m) in the Centre region and;Santchou (latitude 4° 37′ N, longitude 9° 50′ E, rainfall 1500 mm, al-titude 320 m) in the West region of Cameroon. Mature seeds werecollected from at least 30 randomly selected trees per site. Seedsfrom each site were bulked before sowing at the IMPM nursery. Theresulting seedlings were planted near the nursery. These growingseedlings aged one year old were used as cutting source for the pres-ent trial.

2.2. Preparation of cuttings and propagation system

All cuttings taken for the various experimentswere placed in a prop-agation system consisting of poly-propagators (1.5 m × 0.5 m × 1 m)divided into three compartments each. They were constructed ac-cording to the method of Leakey et al. (1990b) and placed in a shadehouse roofed with transparent polyethylene. This provided an irradi-ance level of 20–32% that was received outside the unit.

2.3. Propagation media

The propagation media were composed of fine sand (FS), sawdust(SD) or a fine sand:sawdust (1:1) mixture (FD:SD); all were treatedwith a systemic fungicide (Benlate) prior to use. The sawdust wasobtained from male palm inflorescences. The three media were placedaccording to a randomized block design in the poly-propagators. Basalsingle-node leafy cuttings were collected from Yaounde stock plantsbetween August to September and inserted into a rooting bed in a ran-domized complete block design with four replications. Each treatmentcontained 30 cuttings, giving a total of 360 cuttings for thewhole exper-iment (30 cuttings × 3 treatments × 4 replications). The base of eachcutting was treated with 50 μg IBA. The effect of propagation mediaon cuttings was assessed at three phases: root induction, growth anddevelopment (of aerial parts). Survival and rooting rate, number andlength of roots and number of leaves were measured.

2.4. Growth regulators: IBA and NAA

Four concentrations of IBA and NAA were tested, (control), 50,100, 200 and 300 μg/l. Basal single-node leafy cuttings were collectedfrom Yaounde stock plants in August and September and then,inserted in fine sand in a rooting bed in a randomized completeblock design with four replications. There were 20 cuttings per treat-ment, so the total for the experiment was 720 (20 cuttings × 9 treat-ments × 4 replications). Cuttings were assessed every 2 weeks forsurvival and rooting rate, number and length of roots and numberof leaves.

2.5. Accession site and harvesting period

Single-node cuttings were harvested from each accession plantstock over at three time periods: April to May (CP1); August toSeptember (CP2) and November to December (CP3). The base ofeach cutting was treated with the best growth regulator, as deter-mined from the results of the previous experiment. The cuttingswere inserted in a rooting bed in a randomized complete block designwith four replications. There were 20 cuttings per treatment giving atotal of 720 for the experiment (20 cuttings × 9 treatments × 4 rep-lications). Cuttings were assessed after four months for survival androoting rate, number and length of rooting and number of leaves.

2.6. Accession site and cutting type

For each accession, three types of cutting were produced toevaluate the effect on rooting success: apical cutting (C1), basalsingle-node leafy cutting (C2) and basal two-node leafy cutting(C3). The most appropriate harvest timing, media type and growthregulator concentration, as determined by previous experiment sta-tion were used. Cuttings of different types were inserted in a rootingbed in a randomized complete block design with four replications.Twenty cuttings were used for each treatment; thus the total for theexperiment was 720 (20 cuttings × 9 treatments × 4 replications).Cuttings were assessed after four months for survival, rooting rate,number and length of root and number of leaves.

2.7. Pre-treatment with honey

Basal two-node leafy cutting from Yaounde stock plant was usedto assess the influence of honey on cutting development. Cuttingswere soaked in liquid honey (bought in local market) for a time of 0(control), 30, 60, 90, 120 min. The most appropriate media and phy-tohormone concentration of the previous experiments were equallyused. Cuttings were inserted in a rooting bed in a randomized com-plete block design with four replications. The experimental unit wasmade up of 15 cuttings with a total of 300 (15 cuttings × 5 treat-ments × 4 replications). Cuttings were assessed after two monthsfor survival and rooting rate.

2.8. Substrate used for plant acclimatization

Three types of substrates were used, namely: black soil, fine sand,a mixture of black soil and fine sand (1:1). Plantlets of R. heudelotiiwere placed in polythene pots (10 cm diameter × 18 cm depth).After 2 months of acclimatization, survival rate and plantlet heightwere assessed.

2.9. Data collection and analysis

Data collected were: rate of rooting, number and length of rootsper cutting, number of leaves per cutting, and survival rate. Thesedata were subjected to tests of analysis of variance, correlation andprincipal component analysis. After rejecting null hypothesis ofequal means using ANOVA F-test, Fisher's LSD (Least SignificantDifference) was used for comparing treatment group means at 0.05.Statistical software packages used were SPSS 17.0 and SPAD 4.01 forWindows. Values were means ± standard deviation.

3. Results

3.1. Effect of propagation media on rooting and growth

In the presence of 50 μg IBA, survival rate decreased with time andwas dependent upon media type. During the root induction phase, (0to 4 weeks), survival somewhat decreased, but the effect was not

4 N.D. Tchinda et al. / South African Journal of Botany 88 (2013) 3–9


significant (P b 0.05). In contrast, survival decreased rapidly and sig-nificantly during the growth phase. After 10 weeks of culture (the de-velopment phase), the survival rate stabilized at 54.6%, 45.2% and53.2% for fine sand, sawdust:fine sand and sawdust, respectively. Bysixteen weeks, the percent rooting (53.6; 42.5; 51.2) (Fig. 1A), leafnumber (3.3; 1.1; 5.2) (Fig. 1B), root number (3.5; 0.5; 3.1)(Fig. 1C) and root length (14; 5; 10) (Fig. 1D) were obtained for re-spectively fine sand, sawdust:fine sand and sawdust media. Signifi-cant interaction between sand and sawdust was noticed with leafnumber.

3.2. Effect of IBA and NAA on rooting and growth

The effect of different IBA and NAA concentrations was assessedtwelve weeks after treatment. Cutting survival rate was dependenton treatment. The control had the lowest survival (12.6%); maximumsurvival rates were seen with 100 μg/l NAA (66.5%) and 200 μg/l IBA(61.4%) treatments. Sixteen weeks after culture, the maximum andsignificant (p b 0.05) rooting rate (59.6%) (Fig. 2A), number of leavesper cutting (4.1; 3.9) (Fig. 2B), number of roots (6.4; 4.4) (Fig. 2C) andlength of roots (14.5; 10.9) (Fig. 2D) were obtained, respectively,with 50 or 100 μg/l IBA and NAA. There was a strong positive correla-tion between both survival and rooting rate (R = 0.997) and rootingrate and root number (R = 0.996). Apart from 100 μg/l, IBA and NAAconcentrations did not significantly differ (P b 0.05) between rootingrate and number of leaves per cutting.

3.3. Effect of accessions and harvesting period on rooting and growth

For each accession tested, CP1 (the April to May harvest period)was generally more favorable for rooting and development than ei-ther CP2 (August to September) or CP3 (November to December).The survival rate ranged from 73.2% for cuttings harvested in April–May from the Yaounde accession (A1CP1) to 21.8% for cuttingsharvested in November–December from the Santchou accession(A3CP3). The maximum rate of rooting (68.4%), number of roots(8.6), number of leaves (7.3) and length of roots (14.6 cm) were

obtained for the A1CP1 treatment (Table 1). In general, harvesting pe-riods were significantly different (P b 0.05) from each other andthere was also significant interaction between accessions andharvesting period. Principal component (Jeffer, 1967) showed thatthe first and second components represented 96.94% of the total var-iability. The first axis, which describes 89.5% of total variation, was de-fined by rooting rate. The second axis accounting for 7.45% of totalvariation was defined by leaf number, root number, root length andsurvival rate. Dendrogram produced using average linkage andrescaled distance methods, revealed 3 clusters (Fig. 3):

– cluster I contained cuttings harvested in August–September fromthe Mbalmayo (A2CP2) and Santchou (A3CP2) accessions andwas characterized by the highest survival rate;

– cluster II, which comprised cuttings harvested in August–Septemberfrom the Yaounde accession (A1CP2), and in April to May from theSantchou (A3CP1). Mbalmayo (A2CP1) and Yaounde (A1CP1), wascharacterized by the highest rate of rooting, number of leaves, rootnumber and root length.

– cluster III contained cuttings harvested in November to Decemberfrom the Mbalmayo (A2CP3), Yaounde (A1CP3) and Santchou(A3CP3) accessions; it was not influenced by any variablemeasured.

3.4. Effect of accession and cutting type on rooting and growth

In general, the C1 (apical, single node) cutting type had thehighest survival rate while the C2 (basal, single node) cutting typegave more favorable rooting and development.

Cutting survival rate ranged from 68.9% for basal two-node leafycuttings from the Santchou accession (A3C3) to 86% for apical,single-node leafy cuttings from the Yaounde accession (A1C1). Themaximum rate of rooting (90.6%), root number (8.7), leaf number(7.4) and root length (15.6 cm) were obtained, respectively, for thebasal, single-node leafy cuttings from the Mbalmayo accession(A2C2), basal, single-node leafy cuttings from the Yaounde accession(A1C2), apical single-node leafy cuttings from the Yaounde accession(A1C2) and basal, two-node leafy cuttings from the Santchou

A B

C D

Fig. 1. Effect of propagation media on R. heudelotii cuttings: A) Rooting rate, B) leaf number/cutting, C) root number, D) root length. Values are means ± standard deviation. Valueshaving the same letters are not significantly different according to Fisher's LSD test (P b 0.05).

5N.D. Tchinda et al. / South African Journal of Botany 88 (2013) 3–9


accession (A3C3) (Table 2). In general, accessions and cutting typeand their interaction were significantly different (P b 0.05) fromother treatment; principal component analysis showed that the firstand second components represented 87.47% of the total variabilityobserved. The first, which describes 69.38% of a total variation, wasdefined by the rooting rate, leaf number and, root length variables.The second, 18.09% of total variation comprised root number and sur-vival rate. Average linkage and rescaled distance, techniques pro-duced a dendrogram composed of 4 clusters (Fig. 4):

– cluster I contained apical, single-node leafy cuttings from theMbalmayo and Yaounde accessions (A2C1 and A1C1), basal,single-node leafy cuttings from the Santchou accession (A3C2);it was characterized by the highest survival rooting rates (Fig. 5A);

– cluster II was composed of basal, single-nodes cuttings from theYaounde and Mbalmayo accessions. These materials showed max-imum leaves and longest roots (Fig. 5B);

– cluster III contained basal, two-node leafy cuttings from theMbalmayo (A2C3), Santchou (A3C3) and Yaounde (A1C3) acces-sions; it was characterized by the highest root number (Fig. 5C);

– cluster IV contained apical, single-node leafy cuttings from theSantchou accession (A3C1), was not influenced by any variablemeasured.

3.5. Effect of honey on survival rate and rooting

In general, soaking cutting in honey improved survival and rootingand, with the exception of the longest period, the effect wastime-dependent. The lowest survival (75.9%) and rooting (80%)rates were obtained in the longest time (120 min) treatment, whilethe maximum rate (90.2% and 93.7%, respectively) were obtainedwith 60 min of soaking (Fig. 6).

3.6. Effect of substrate on plant acclimatization

Plants derived from cuttings were potted on substrate containingblack soil, fine sand or a 50:50 mixture of black soil:fine sand. After6 weeks of acclimatization, the 50:50 mixture showed a success rate(67.8%) which was significantly different (P b 0.05) from that of theother substrates (49.9%). Plantlets had a mean height of 24.7, 19.4and 16.2 cm respectively; in the 50:50 mixture, fine sand and blacksoil.

4. Discussion

The most effective means of improving R. heudelotii for on-farmuse is by vegetative propagation, which can rapidly overcome the

C D

A B

0

Fig. 2. Effect of IBA ■ or NAA □ concentration on R. heudelotii cuttings. A) Rooting rate, B) leaf number, C) root number, D) root length. Values are means ± standard deviation.Values having the same letters are not significantly different according to Fisher's LSD test (P b 0.05).

Table 1Effect of accession and harvest time on rooting and growth of Dacryodes edulis after 16 weeks of culture.

Accessions Harvest period Survival rate Rooting rate Number of roots/cutting Number of leaves/cutting Length of roots/cutting

Yaounde (A1) CP1 73.2 ± 4.2 d 68.4 ± 3.4 f 8.6 ± 0.8 e 7.3 ± 0.3 d 14.6 ± 1.8 eCP2 66.5 ± 3.1 b 59.6 ± 2.3 d 8.4 ± 0.6 d 6.1 ± 0.8 bc 14.3 ± 1.5 eCP3 24.6 ± 0.9 a 22.2 ± 1.0 b 5.2 ± 0.4 ab 5.2 ± 0.5 b 10.9 ± 1.0 bc

Mbalmayo (A2) CP1 70.9 ± 3.9 c 63.5 ± 1.5 e 6.8 ± 0.6 c 6.5 ± 0.5 c 12.1 ± 0.9 cdCP2 65.6 ± 2.8 b 52.0 ± 2.0 c 6.1 ± 0.9 b 6.9 ± 0.9 c 10.2 ± 0.8 bCP3 22.0 ± 0.7 a 22.0 ± 0.6 b 4.2 ± 0.3 a 4.2 ± 0.3 a 7.0 ± 0.8 a

Santchou (A3) CP1 68.9 ± 1.9 c 58.9 ± 1.4 d 7.2 ± 1.0 c 6.3 ± 0.5 c 11.0 ± 1.1 cCP2 65.8 ± 2.3 b 50.0 ± 1.1 c 6.3 ± 0.6 bc 6.5 ± 0.8 c 11.5 ± 0.9 cCP3 21.8 ± 0.4 a 17.8 ± 0.4 a 4.0 ± 0.3 a 4.5 ± 0.2 a 8.6 ± 0.7 a

Values having the same letter(s) in the same column are not significantly different according to Fisher's LSD Test at P b 0.05.CP1: The April to May harvest period of cuttings; CP2: The August to September harvest period of cuttings; CP3: The November to December harvest period of cuttings



limitations imposed by long generation times, irregular fruiting/flowering and out breeding. In this work, multiplication was achievedin non-mist propagation system, confirming results of Shiembo et al.(1997a). The fact that maximum survival and rooting percentages of90.2 and 93.7%, respectively, were achieved in all five experimentssuggests that the technique described here could be used forlarge-scale multiplication of the species. Previous trials of Shiemboet al. (1997a) assessed the effect of propagation medium, auxin con-centration and leaf area on rooting of leafy stem cuttings; only 80%rooting was achieved, compared to over 90% in this study.

We observed that rooting is generally dependent upon pre-treatment of the cutting (with growth regulator or honey), pottingmedia, cutting type and harvesting time. The highest rooting and sur-vival percentage were observed in the fine sand medium. These re-sults correspond with those of Leakey et al.(1990b), who obtainedmaximum rooting from cuttings of Cordia alliodora and Gmeliaarborea by using fine sand. Shiembo et al. (1997a) showed that forthe propagation by cuttings, fine sand is superior to sawdust:finesand, sawdust:gravel or fine sand:gravel. In contrast, experimentswith Vitex madiensis (Mapongmetsem, 2006) or Vochysia hondurensis(Mesen, 1993) showed that a 50:50 mixture of sawdust:sand is favor-able for propagating these species by cuttings. Different species maydisplay contrasting rooting percentages in different rooting mediawhich may be attributed to variation in oxygen and water content,pH or porosity of the media while survival rate of cuttings may beinfluenced by media humidity and inherent rooting capacity(Leakey, 1990a). In general, sand is an appropriate rooting medium,with an optimal volume of gas-filled pore-space and oxygen diffusionrate adequate for the needs of respiration (Andersen, 1986). Also, itlimits development of microbial pathogens.

IBA had a stronger effect on survival and rooting rate than NAA.IBA has been used for rooting of many tropical trees such as Miliciaexcelsa (Ofori et al., 1996), C. alliodora (Mesen et al., 1997), Prunus

sp. (Esitken et al., 2003; Tchoundjeu et al., 2002a, 2002b); Shoreaparvifolia and Shorea macroptera (Aminah et al., 2006) and Dalbergiamelanoxylon (Amri et al., 2010). For Cordia alliodora, an IBA concen-tration of 1.6% produced the highest rooting percentage (Mesen etal., 1997) and cuttings without IBA failed to root (Mesen, 1993). How-ever, in M. excelsa, a decline in rooting percentage with IBA concen-trations greater than 0.2% was reported (Ofori et al., 1996).Successful rooting without applied auxin has also been reported in anumber of other tropical tree species, such as Shorea macrophylla(Lo, 1985), Nauclea diderrichii (Leakey, 1990a) and Vochysiahondurensis (Leakey et al., 1990b). It is known that concentrationsof auxin substantially higher than those normally found in plant tis-sues may cause cell death (Hartmann et al., 1990). In this study, eitherIBA or NAA concentrations greater than to 100 μg/l reduced rootingrates, as well as the number of leaves and roots and root-length.

Accession source had no significant effect on rooting and cuttinggrowth but harvesting period is of importance. In contrast,Somkuwar et al. (2011) described rootstock variability in the abilityfor rooting based on the biochemical constituents of the Vitis sp. Inthis experiment, the April–May period, which corresponds to the be-ginning of raining season, is generally more favorable for rooting anddevelopment of cutting than either theAugust–September orNovember–December period, which correspond to the peak rainy and end-of-rainyseason, respectively. Generally, each period is characterized by specificphysiological and biochemical stages. Cuttings taken in April–May andAugust–September were in a vegetative growth state while thoseharvested in November–December were dormant. In contrast, Auge(1989) demonstrated that dormant cuttings are more responsive thanactively-growing cuttings. In Hevea brasiliensis, which exhibits rythmicgrowth independent of seasonal cycle, cuttings harvested during the veg-etative or active phase are most suitable for rooting and development(Lardet et al., 1990). Harvest period had a strong effect (68.4% of total)

A1CP3

A3CP3

A2CP3

A1CP1

A2CP1

A3CP1

A3CP2

A2CP2

A1CP2

510152025 0 35 30

Fig. 3. Dendrogram of 3 accessions and cutting harvest period of R. heudelotii based onrescaled distance cluster combination of 5 morphological parameters.

Table 2Effect of accession and cutting type on rooting and growth of Dacryose edulis, after 16 weeks of culture.

Accessions Type of cutting Survival rate of cutting Rate of rooting Number of roots/cutting Number of leaves/cutting Length of roots/cutting

Yaounde (A1) C1 86.0 ± 3.4 d 80.0 ± 3.0 e 8.3 ± 0.6 c 7.1 ± 0.8 b 12.6 ± 1.2 bC2 80.3 ± 3.2 bc 89.6 ± 2.8 f 8.7 ± 0.7 c 7.4 ± 0.4 c 12.1 ± 0.9 bC3 73.2 ± 2.8 ab 68.4 ± 1.5 c 8.6 ± 0.9 bc 7.3 ± 0.6 c 14.6 ± 1.5 c

Mbalmayo (A2) C1 84.3 ± 3.8 d 80.1 ± 1.9 e 7.6 ± 0.7 b 7.0 ± 0.9 b 14.5 ± 0.8 cC2 75.6 ± 2.7 b 90.6 ± 2.9 f 8.1 ± 1.0 b 6.9 ± 0.3 b 11.0 ± 0.7 aC3 70.9 ± 2.6 a 63.5 ± 1.7 b 6.8 ± 0.5 a 6.5 ± 0.7 a 15.1 ± 1.3 d

Santchou (A3) C1 82.9 ± 3.1 c 65.1 ± 0.9 b 6.9 ± 0.6 a 6.8 ± 0.4 b 10.3 ± 1.0 aC2 78.3 ± 1.9 b 75.6 ± 1.4 d 6.8 ± 0.8 a 6.4 ± 0.8 a 10.9 ± 0.8 aC3 68.9 ± 1.5 a 58.9 ± 0.8 a 7.2 ± 0.6 a 6.3 ± 0.3 a 15.6 ± 1.6 d

Values having the same letter(s) in the same column are not significantly different according to Fisher's LSD Test at P b 0.05.C1: Apical cutting; C2: Basal single-node leafy cutting; C3: Basal two-node leafy cutting.

510152025 0 35 30

A1C1

A2C1

A3C2

A3C1

A1C3

A3C3

A2C3

A2C2

A1C2

Fig. 4. Dendrogram of 3 accessions and cutting type of R. heudelotii based on rescaleddistance cluster combination of 5 morphological parameters.



on rooting rate. Rooting being a prerequisite for continued growth anddevelopment of plants, this factor is then very important for propagationof R. heudelotii.

Cutting type also had significant effects on rooting independent ofthe accession. It is responsible for almost 70% of observed results forthree parameters measured: (rooting rate, leaf number and rootslength). Undifferentiated tissues of woody plants are generally moreresponsive than differentiated structures (Auge, 1989). Additionally,response was enhanced when the cuttings were soaked for 60 minin honey, possibly because the honey provides protection against des-iccation and rotting, which are the main causes of mortality (Shiemboet al., 1997a). Honey which is natural and non toxic compared toother substances like petroleum and silicon also used to prevent des-iccation could be suitable to treat cuttings. After acclimatization, max-imum survival rate of plants was 67.6%; these results are similar tothose obtained by Youmbi and Benbadis (2001) for D. edulis andFotso et al. (2004) for R. heudelotii, who used the same substrate foracclimatization as reported here. This work provides an improvedprotocol which can be used for propagation of R. heudelotii in agrofor-estry systems.

Acknowledgments

This research was supported by the International Foundationfor Sciences (IFS), Sweden, through Grant No.: D/4365 received by

Dr. Nehemie Donfagsiteli Tchinda. We thank Pamela Moon; TropicalResearch and Educational Center; University of Florida, for her assis-tance in the correcting this manuscript.

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A B C

Fig. 5. Morphology of R. heudelotii cuttings. A) Apical, single-node leafy cuttings with a well developed root system, B) basal, single-node leafy cuttings demonstrating the greatestnumber of leaves and roots, C) basal, two-node leafy cuttings, demonstrating greatest root length (lines = 1.5 cm).

Fig. 6. Variation survival and – – rooting rates of cuttings depending by honey timetreatment, after 8 weeks of culture.



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Improving propagation methods of Ricinodendron heudelotti Baill. from cuttings

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