Available online at http://www.ifgdg.org Int. J. Biol. Chem. Sci. 17(3): 996-1005, April 2023 ISSN 1997-342X (Online), ISSN 1991-8631 (Print) Original Paper http://ajol.info/index.php/ijbcs http://indexmedicus.afro.who.int Root segments cuttings of Vitex doniana Sweet: effect of origin and alignment Guidawa FAWA1*, Benjamine ANGUESSIN2, Jean Baptiste BINWE1, Zéphirin OUMAROU3 and Pierre Marie MAPONGMETSEM1 1 Department of Biological Sciences; University of Ngaoundéré, Faculty of Science, Laboratory of Biodiversity and Sustainable Development; P.O Box: 454 Ngaoundéré, Cameroon. 2 Department of Environmental Sciences, National Polytechnic School of Maroua University of Maroua; BP: 46 Maroua, Cameroon. 3 Department of Plant Sciences, Faculty of Science, The University of Bamenda; P.O Box: 39 Bambili, Cameroon. *Corresponding author; E-mail: fawaguidawa@gmail.com. Received: 02-01-2023 Accepted: 04-04-2023 Published: 30-04-2023 ABSTRACT Vitex doniana (Verbanaceae) is one of the most popular local species among farmers in the high Guinean savannah of Cameroon. Unfortunately, it is under anthropogenic pressure and this tures to reduce its population. The objective of this work was to study the effect of the origin and the alignment of the cuttings of the root segments on budding and rooting. The origin of the cuttings (Dang, Poli) constituted the main treatment and their alignment in the substrate represented the secondary treatment. The cutting of the root segments was carried out according to an experimental device in split-plot with three replications. The experimental unit consists of 10 cuttings. A total of 180 cuttings of root segments (2×3×3×10) were handled. It emerged from this study that cuttings from root segments of different origins reacted differently to vegetative propagation by cuttings. Cuttings from Poli budded better (67.77±26.82%) than those from Dang (36.66±12.24%). The analysis of variance showed a significant difference (0.0001˂0.001) with regard to the origin of the cuttings. The alignment of the cuttings in the substrate also affected budding. The vertical and oblique alignment had the best budding rates (63.33%). The buds appeared mainly at the proximal pole of the cuttings (44.44±26.6%). As for rooting, the cuttings from Poli took better root (57.41±9.32%) than those from Dang (26.35±6.2%). These results contribute to the optimization of the domestication strategies of this species. The domestication of this species of socio-economic interest could contribute to improving the living conditions of local populations. © 2023 International Formulae Group. All rights reserved. Keywords: Vitex doniana, Origin, alignment, Cuttings, Root, Substrate. INTRODUCTION The tropical dry forests constitute an exceptional reservoir of biodiversity. They provide ecosystem goods (fruit, fodder, medicine, etc.) and ecological services (Schreckenberg et al., 2006). The degradation of this biodiversity generated by anthropogenic activities gives rise to a feeling of concern at the environmental, social and © 2023 International Formulae Group. All rights reserved. DOI : https://dx.doi.org/10.4314/ijbcs.v17i3.19 economic consequences that could result from it (Balvanera et al., 2006; Cardinale, 2012). In Cameroon in general and in the Northern part in particular, ecosystems are diversified and rich in species of socioeconomic interest with plants such as Vitex doniana (Verbanaceae), which are among the most popular local species to peasants 9348-IJBCS G. FAWA et al. / Int. J. Biol. Chem. Sci. 17(3): 996-1005, 2023 (Tchiégang-Megueni et al., 2001; Mapongmetsem and Laissou, 2010). Indeed, V. doniana is a tree that is widely used in several regions of Africa for its multiple qualities. Unfortunately, this species still lives in the wild and is under various pressures (Mapongmetsem et al., 2000). Given its importance to rural communities, this plant deserves more attention in order to maximize the potential it represents. Therefore, the low cost vegetative propagation technique such as cuttings of root segments appears as one of the viable solutions for the multiplication of endangered species (Meunier et al., 2008, Mapongmetsem et al., 2012). It makes the exact copy of the parental characters and has advantages following the production of plants identical to the parental generation, the rapid production of plants, especially in species whose multiplication by seed is difficult or poor (Hannah and Jan, 2003). A perfect mastery of both exogenous and endogenous factors will facilitate the multiplication of this plant by cuttings under polypropagator. Several studies carried out with this in mind (Mapongmetsem et al., 2016; Fawa, 2015) have focused on various aspects. The lack of scientific information on the origin and alignment of cuttings of root segments in the substrate is a major handicap. Hence the work objective which was to study the effect of the origin and the alignment of the cuttings of root segments in the substrate on budding and rooting. Cameroon. Poli belongs to this zone, the prevailing climate is of the Sudanian type with a humid nuance. It is characterized by two distinct more or less contrasting seasons. A dry season ranging from 6 to 7 months from November to April, a rainy season which extends over 4 to 5 months from May to October. The average temperature ranges from 27°C for the coolest months to 32°C for the hottest months. The soil is made up of ferralitic structures, hydromorphs and planosols with thick horizons. The vegetation looks like a clear forest reminiscent of the plant formations of the Adamawa plateau. The most common ligneous species are: Isoberlinia doka, Anogeissus leiocarpa, Terminalia laxiflora, Piliostigma thonningii (PDC, 2014). Sampling design The removal of the Vitex doniana root fragments consisted in carefully excavating a few carefully chosen mother plants (healthy individuals, present tree), from the first hours of the morning to minimize cell dehydration. The fragments of lateral roots taken were cut using a 20cm cutting pruner, a dimension favorable for the budding of Vitex doniana root (Mapongmetsem et al., 2017). The root segment cuttings were brought to the nursery using a cooler containing pieces of ice. At the nursery, the root segment cuttings from Dang and Poli were cultivated in the polypropagagator, containing previously prepared substrate composed of the black earth/sawdust mixture (50/50) which is best for budding the cuttings of root segments of Vitex doniana (Mapongmetsem et al., 2012). These roots segments cuttings were arranged in a vertical, horizontal and oblique alignment. The test was watered daily morning and evening using a sprayer which delivered the water in very fine droplets. The evaluation of the test was carried out weekly as soon as the first buds appeared. MATERIALS AND METHODS Description of Study site Sampling of Vitex doniana root segment cuttings took place in two agroecological zones: the highland Guinean savannah zone more precisely in Dang and the Sudano-Sahelian zone more precisely in Poli. The Eastern Sudano-Sahelian zone includes between 8°36" to 12°54" north latitude, and 12°30" to 15°42" east longitude. It covers the North and Far North regions of 997 G. FAWA et al. / Int. J. Biol. Chem. Sci. 17(3): 996-1005, 2023 The experimental device used is splitplot with three replication, in which the origin of the cuttings (Dang, Poli) constitutes the main treatment and the alignment of the cuttings in the substrate represents the secondary treatment. The experimental unit consisted of 10 cuttings. A total of 180 roots segment cuttings (2×3×3×10) were handled. alignment at 63.33 ± 22.5% and 63.33 ± 28.04% in those inserted in a vertical and oblique alignment (Figure 2). The analysis of variance indicated a significant difference (0.0004 ˂ 0.001). The alignment of the cuttings of root segments influenced budding. Effect of the interaction between the origin * alignment of the cuttings of root segments Concerning the interaction between the origin*alignment of the root segment cuttings, at the end of the test, budding rate varied from 23.33 ± 5.65% in cuttings of root segments from Dang inserted in the substrate in alignment horizontal at 86.7 ± 15.27% in those of Poli, inserted into the substrate in an oblique alignment (Table 1). The analysis of variance did not indicate a significant difference (0.0855 ˃ 0.05). The origin and alignment of the combined cuttings did not influence the budding of the cuttings of root segments. Data collection and statistical analysis Data collection was done weekly when the first buds appeared. The number of cuttings having budded, pole of appearance of buds, the number of leaves, the number of aerial axes, the height of the aerial axes, the number of cuttings having emitted roots, the number of roots, the length of the root were evaluated. The data collected was analyzed using Statgraphic plus 5.0 software which was used to perform the analysis of variance. Duncan Multiple Range's Test was used to separate the significant means. Polarity of buds Origins effect on parts of bud appearance In general, the rate of appearance of buds on cuttings of root segments varied from 0.55 ± 0.39% at the distal pole to 44.44 ± 26.6% at the proximal pole (Table 2). The pole of appearance of the buds at the proximal pole obeys the budding of the cuttings of root segments. The analysis of variance showed a significant difference for the buds that appeared at the proximal pole (0.0039 ˂ 0.01) and on the middle part (0.0279 ˂ 0.05) of the cuttings from the two localities. Influence of the insertion of cuttings in the substrate on the appearance of buds Regarding the arrangement of the cuttings of root segments in the substrate, the rate of appearance of buds varied from 0.5 ± 0.32% at the distal pole to 44.22 ± 18.71% at the proximal pole (Table 3). The cuttings inserted vertically into the substrate had a budding rate of 61 ± 23.16% at the proximal pole. The analysis of variance presented a significant difference for the proximal pole RESULTS Budding of root segments cuttings The cuttings of Vitex doniana root segments were cultivated on August 01, 2018 and the first aerial shoots appeared on September 10, 2018, 6 weeks after their cultivation. Effect of origin of cuttings Eighteen weeks after the cuttings were cultivated, the budding rate varied from 36.66 ± 12.24% in root segment cuttings from Dang to 67.77 ± 26.82% in those from Poli (Figure 1). The analysis of variance indicated a significant difference between the origins (0.0001˂0.001). Effect of cuttings alignment Concerning the alignment of the root segment cuttings in the substrate, the budding rate oscillates between 30 ± 8.94% in roots segments cuttings inserted in horizontal 998 G. FAWA et al. / Int. J. Biol. Chem. Sci. 17(3): 996-1005, 2023 (0.0001 ˂ 0.001) and the middle part (0.023 ˂ 0.05) for the three types of insertion of cuttings into the substrate. alignment in the substrate to 53.35 ± 7.20% and 50. 31 ± 11.2% inserted respectively in an oblique and vertical alignment in the sand/sawdust substrate (Figure 5). The analysis of variance indicated a significant difference (0.002˂0.01). The oblique alignment in the substrate improved the rooting rate of the cuttings. Rooting cuttings from root segments At the end of the trial, at 18 weeks after their cultivation, several cuttings of root segments from all positions and from all origins had taken roots (Figure 3). Influence of the origin interaction * alignment of the cuttings in the substrate The rooting assessment occurred at the end of the trial that is to say at the 18th week. The percentage of cuttings rooted according to the source and the method of insertion in the sand/sawdust substrate varies from 21.34±5.77 in cuttings inserted in a horizontal alignment for cuttings from Dang to 83.61±11.27 in those inserted in an oblique alignment for cutting originating in Poli (Table 4). The analysis of variance shows no significant difference (0.085 ˃ 0.05). The alignment of the cuttings did not influence the rooting rate. Influence of the origin of the cuttings At the 18th week, the percentage of cuttings of root segments rooted according to origin varied from 26.35 ± 6.2% for cuttings of root segments from Dang to 57.41 ± 9.32% for those from Poli (Figure 4). The analysis of variance indicated a significant difference (0.026 ˂ 0.05). The origin of the cuttings influenced rooting. Effect of the alignment of the cuttings in the substrate At the end of the trial (18th week), the rooting rate fluctuated from 26 ± 8.94% in root segment cuttings inserted in a horizontal Figure 1: Variation of the percentage of budding according to the origin of the cuttings of the root segments. 999 G. FAWA et al. / Int. J. Biol. Chem. Sci. 17(3): 996-1005, 2023 Figure 2: Percentage of budding depending on the alignment of the root segment cuttings in the substrate. Table 1: Rate of budding as a function of the origin and alignment of the cuttings in the substrate. Origin of cuttings Poli Dang Vertical 80±12.70% 46.66±2.78% Alignment of cuttings in the substrate Horizontal Oblique 36.66±4.35% 86.66±15.27% 23.33±5.65% 40±9.10% Table 2: Rate of appearance of buds on the different parts of cuttings according to origins. Origin of cuttings Dang Poli Means Probability PP 32.22±17.15a 52.66±36.05b 44.44±26.6 0.0039 Part of appearance of buds on cuttings MP PPM 2.22±1.57a 2.22±1.21 7.77±6.66b 2.22±1.36 4.99±4.1 2.22±1.57 0.0279 1.000 DP 00±00 1.11±0.78 0.55±0.39 0.3370 Means followed by the same letter are statistically identical (p < 0.05). PP= Proximal pole, MP= Middle part, PPM= Proximal pole and Middle part, DP= Distal pole. Table 3: Rate of budding according to the polarity of the cuttings according to the alignment. Part of appearance of buds on cuttings MP PPM Alignment of the cuttings PP Horizontal 13.33±5.16a 10±6.32c 5±2.5 1.66±0.96 Oblique 58.33±27.83b 3.33±1.92b 1.66±1.66 00±00 Vertical 61±23.16c 1.66±1.92a 00±00 00±00 Means 44.22±18.71 4.99±3.38 2.22±2.22 0.5±0.32 Probability 0.0001 0.023 0.1393 0.3966 Means followed by the same letter are statistically identical (p < 0.05). PP= Proximal pole, MP= Middle part, PPM= Proximal pole and Middle part, DP= Distal pole. 1000 DP G. FAWA et al. / Int. J. Biol. Chem. Sci. 17(3): 996-1005, 2023 aa b Figure 3: Cuttings of root segments with adventitious roots depending on the alignment (vertical cutting (a); horizontal cutting (b)). b a Figure 4: Percentage of rooting of cuttings according to their origins. 1001 G. FAWA et al. / Int. J. Biol. Chem. Sci. 17(3): 996-1005, 2023 a a b Figure 5: Percentage of rooting according to the alignment of the cuttings in the substrate. Table 4: Rooting of cuttings depending on the origin and alignment of the cuttings. Origin of cuttings Alignment of cuttings in the substrate Vertical Horizontal Oblique Dang 38.66±5.32% 21.34±5.77% 37.39±6.29% Poli 70±2.46% 33.86±2.48% 83.61±11.27 DISCUSSION Budding of cuttings The budding time in this species was 6 weeks. Similar results are reported on Lophira lanceolata (Fawa, 2015) and on Vitex doniana (Mapongmetsem et al., 2016) in the same locality as ours. The origin significantly influenced the budding rate. Root segments tend to bud more depending on their ecological origin. This situation could be explained by the fact that budding is influenced by the interaction of endogenous and environmental factors (Tchoundjeu et al., 2002). Dang and Poli belong to different agroecological zones. Along the same line, Boutherin and Bron (2002) reported that budding depends on the genotype of each individual, the period of setting up the trial, the age and the physiological state of the individuals. This ability is believed to be related to the period of carbohydrate accumulation in Poli cuttings. These carbohydrates play an important role in the budding process of cuttings and the growth of newly formed adventitious buds as reported by Akouethe et al. (2014) on three species: Haematostaphis bateri, Lannea microcarpa and Sclerocarya birrea. These results also flow in favor of the ecological and soil conditions of each cutting area. 1002 G. FAWA et al. / Int. J. Biol. Chem. Sci. 17(3): 996-1005, 2023 The alignment of the cuttings of root segments influence the budding. The result obtained in this species was similar to that of Ghani and Calahan (1991), on Prunus avium. They reported that the vertical alignment is best for budding. In Burkina Faso, KyDembélé et al. (2010) reported similar results on Detarium microcarpum. Indeed, when the cutting is inserted vertically or obliquely in the substrate, 1 cm from the proximal end is exposed. In our case, exposure would be the exogenous factor that would have most influenced budding. The appearance of buds or roots is very often polarized, the pole located near the end of the roots will rather induce roots while the pole located in the direction of the collar will induce the buds (Boutherin and Bron, 2002). The pole of appearance of the buds in the proximal alignment obeys the budding of the cuttings of root segments. In the Guinean savannah highlands, Oumarou et al. (2018) reported similar results on Securidaca longepedunculata. In Burkina Faso, Harivel et al. (2006) showed that 90% of leafy shoots develop at the proximal end and adventitious roots at the distal end in Faidherbia albida. which by photosynthesis produce the carbohydrates which favors the initiation of the roots. Conclusion The aim of this work was to study the effect of the origin and alignment of cuttings from root segments on the ability to neoform buds and adventitious roots. It turned out that Vitex doniana has good new formation skills for buds and adventitious roots. The cuttings of root segments from different origins react differently to vegetative propagation. Cuttings from Poli buded better than those from Dang. The alignment of the cuttings of root segments in the substrate also affected budding and rooting. The oblique and vertical alignment are those which are best suited for testing cuttings. The buds appeared mainly at the proximal pole of the cuttings of root segments. Cuttings from Poli rooted better than those from Dang. The vertical and/or oblique alignment promoted the rooting of cuttings from root segments. COMPETING INTERESTS The authors confirm that there is no conflict of interest for the publication of this article. They all agree that it be published in this journal. Rooting cuttings The origin of the cuttings of root segments influenced the roots. This is also the result of the work of Hussain et al. (2018) on the cuttings of Rubus spp. collected at different times of the year. The development of adventitious roots is slow compared to the emission of buds. In fact, the buds produced the sugars and hormones responsible for the formation of adventitious roots through the process of photosynthesis (Mapongmetsem et al., 2017; Ky-Dembélé et al., 2010). The synthesized carbohydrates circulate from the leaves through the sap produced towards the distal pole of the cuttings where they induce the formation of adventitious roots. The oblique alignment in the substrate favors the rooting of the cuttings. The alignment of cuttings in the substrate influences the growth of adventitious roots. Wachowski et al. (2014) reported that the new formation of the roots is dependent on the emergence of leafy shoots AUTHORS’ CONTRIBUTIONS GF: Took part in the design of the study and the production; Analysis and interpretation of the results. BA: Took part in the field work and laboratory. JBB: Took part in the field work and laboratory. ZO: Took part in the field work and laboratory. PMM: Took part in the design of the study and the production; Analysis and interpretation of the results; Project designer. ACKNOWLEDGEMENTS The authors would like to thank the reviewers for their contributions which improved the quality of the article. REFERENCES Akouethe A, Bammite D, Tozo K, Akpagana K. 2014. Contribution a la Multiplication 1003 G. FAWA et al. / Int. J. Biol. Chem. Sci. 17(3): 996-1005, 2023 DOI: https://doi.org/10.1590/010029452017939 Ky-Dembélé C, Tigabu M, Bayala J, Savadogo P, Boussin IJ, Odem P. 2010. Clonal propagation of Detarium microcarpum from root cuttings. Silva Fennica, 44(5): 775-787. DOI: https://doi.org/10.14214/sf.452 PCD. 2014. Plan Communal de Développement de Ngaoundéré III, p. 249. Mapongmetsem PM, Kapchie VN, Tefempa BH. 2012. Diversity of Local Fruit Trees and their Contribution in Sustaining the Rural Livelihood in the Northern Cameroon. Ethiopian Journal of Environmental Studies and Management, 5(1): 32-46. DOI: 10.4314/ejesm.v5i1.5 Mapongmetsem PM, Mbofung G, Ibrahima A, Tchuenguem Fohouo FN, Alexandre DY, Mefeng O. 2000. Situation et Dynamique des Jardins de case dans les Savanes Soudano-guinéennes de l’Adamaoua : Cas de la Zone Périurbaine de Ngaoundéré (Adamaoua, Cameroun). Biosciences Proceed, 7: 403-412. DOI: https://doi.org/10.19044/esj.2022.v18n27 p227 Mapongmetsem PM, Sonou AP, Raouguedam J, Koye LB, Fawa G. 2016. Vegetative propagation of Sclerocarya birrea (A. Rich.) Hochst. From Root Segments Cuttings: Effect of Substrate and Root Diameter. Annals of Experimental Biology, 4(2): 23-32. Mapongmetsem PM, Laissou M. 2010. Contribution a la Domestication des Fruitiers Locaux : Influence des Substances de Croissance et des Substances de Croissance sur L’enracinement des Marcottes. Com. International Symposium, Indigenous Fruit Trees for Dryland Africa: Domestication for Use in a Changing Environment. Allada, Benin, p. 12. Mapongmetsem PM, Djomba E, Fawa G, Oumarou Z, Dangai Y, Bellefontaine R. 2017. Vegetative Propagation of Vitex doniana Sweet from Root Segments Cuttings: Effects of Substrate and Par Graines Et Par Bouturage de Segment de Tiges et de Racines, de Trois Fruitiers Spontanés de la Région des Savanes au Togo: Haematostaphis barteri hook. f., Lannea microcarpa Engl. & K. Krauss et Sclerocarya birrea (A. Rich) Hochst. European. Scientific Journal, 10(6): 195-211. DOI: https://doi.org/10.19044/esj.2014.v10n6p %25p Balvanera P, Pfisterer AB, Buchmann N, He JS, Nakashizuka T, Raffaelli D, Schmid B. 2006. Quantifying the Evidence for Biodiversity Effects Ecosystem Functioning and Services. Eco. Lett., 9: 1146-1156. DOI: 10.1111/j.14610248.2006.00963.x Boutherin D, Bron G, 2002. Multiplication des Plantes Horticole (2e edn). Edition Lavoisier, TEC et Doc ; p. 204. Cardinale BJ. 2012. Impacts of biodiversity loss. Science, 336(6081): 552-553. DOI:10.1126/science.1222102 Fawa G. 2015. Phénologie et mode de propagation des trois espèces agroforestiers locales des hautes savanes guinéennes (Adamaoua, Cameroun). Thèse de Doctorat /Ph.D. Uni Cameroun, p. 171. Ghani AKMO, Cahalan CM. 1991. Propagation of Prunus avium from root cuttings. Forestry, 64(4): 403-409. DOI: https://doi.org/10.1093/forestry/64.4.40 3 Hannah J, Jan B. 2003. Multiplication Végétative des Ligneux en Agroforesterie Manuel de Formation et Bibliographie. World Agroforestry Center (WAC), Naïrobi, p. 162. Harivel A, Bellefontaine R, Ousmane B. 2006. Aptitude à la Multiplication Végétative de Huit Espèces Forestières d’intérêt au Burkina Faso. Bois et Forêts des Tropiques, 288(2): 39-50. Hussain H, Ruffo RS, Carlos Colombo R, Marinhode Assis AR. 2018: Cutting types Collected at Different Season on blackberry multiplication. Rev. Bras. Fructic., 39(3): 29-39. 1004 G. FAWA et al. / Int. J. Biol. Chem. Sci. 17(3): 996-1005, 2023 Length of Cuttings on the Rooting Ability. Annals of Experimental Biology, 5(1): 18-24. Meunier Q, Bellefontaine R, Monteuuis O. 2008. La Multiplication Végétative d’arbre et d’arbuste Médicinaux au Bénéfice des Communautés rurale d’Ouganda. Bois et Forêts des Tropiques, 295(2): 71-82. Oumarou ZH, Hamawa Y, Tsobou R, Abdoulaye H, Bellefontaine R, Mapongmetsem PM. 2018. Multiplication Végétative de Securidaca longepedunculata Fresen par Bouturage de Segments de Racine. Afrique Science 14(6): 388 – 399. Schreckenberg KE, Marshall A, Newton D, Velde WT, Rushton J, Edouard F. 2006. Commercialization of Non-Timber Forest Products: What Determines Success? ODI Forestry Briefing, 10: 1-6. DOI: https://odi.org/en/publications/commerci alisation-of-non-timber-forest-productswhat-determines-success/ Tchiégang-Megueni C, Mapongmetsem PM, Akagou Zedong C H, Kapseu C. 2001. An Ethnobotanical Study of Indigenous Fruit Trees in Nothern Cameroon. Forests, Trees and Livelihoods, 11: 149158. DOI: https://doi.org/10.1080/14728028.2001.9 752382 Tchoundjeu Z, Avana ML, Leakey RRB. 2002. Vegetative Propagation of Prunus africana: Effects Of Rooting Medium, Auxin Concentrations and Leaf Area, Agroforestry Systems, 54(3): 183–192. DOI: 10.1023/A:1016049004139 Wachowski J, Landhäusser SM, Lieffers VJ. 2014. Depth of Root Placement, Root Size and Carbon Reserves Determine Reproduction Success of Aspen Root Fragments. Forest Ecology and Management, 313: 83–90. DOI: http://dx.doi.org/10.1016/j.foreco.2013.1 1.003 1005