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Article

Effect of Propagation Techniques on Growth, Development, Oil Yield, and Quality of Medicinal Cannabis (Cannabis sativa) Found in Lusikisiki, Eastern Cape, South Africa

by
Azile Dumani
1,2,*,
Tembakazi Theodora Silwana
2,
Ifeanyi Moses Egbichi
1,*,
Adebola Omowunmi Oyedeji
3,
Babalwa Mpambani
2 and
Hlabana Alfred Seepe
4
1
Department of Biological and Environmental Sciences, Faculty of Natural Science, Walter Sisulu University, Mthatha 5117, Eastern Cape, South Africa
2
Department of Rural Development and Agrarian Reform, Döhne Agricultural Development Institute, Stutterheim 4930, Eastern Cape, South Africa
3
Department of Chemical and Physical Sciences, Faculty of Natural Science, Walter Sisulu University, Mthatha 5117, Eastern Cape, South Africa
4
Department of Agriculture, Land Reform & Rural Development, Quarantine Station, Polkadraai Road, Stellenbosch 7600, Western Cape, South Africa
*
Authors to whom correspondence should be addressed.
Horticulturae 2025, 11(12), 1428; https://doi.org/10.3390/horticulturae11121428
Submission received: 3 August 2025 / Revised: 10 October 2025 / Accepted: 18 October 2025 / Published: 26 November 2025
(This article belongs to the Section Medicinals, Herbs, and Specialty Crops)
Browse Figures
Versions Notes

Abstract

This study investigated the influence of cutting techniques on the growth, development, yield, and oil quality of Cannabis sativa found in the Eastern Cape Province. The greenhouse pot experiment was conducted at Dohne Agricultural Development Institute (DADI), Stutterheim, Eastern Cape, during the winter and summer growing seasons of 2024/25. It was laid out in a Randomized Complete Design (RCD) with three treatments replicated three times. The treatments used were herbaceous shoot cutting with two different leaf area (LA) trimming amounts and sexual propagation. The parameters measured were plant height, number of branches, stem girth, number of weeks to first flowering, number of flowers, flower sex, number of weeks to 50% embar colorations, plant fresh weight, leaf and flower weights, and dry leaf and flower weights. The flower oil yield and cannabinoid composition were determined using GC-MS. The results indicate that the sexually propagated plants were taller (p < 0.05) with vigorous growth; had the highest fresh plant, leaf, and dry leaf weights; and had a higher number of male flowers overall. Herbaceous shoot cutting without LA trimming showed a significantly higher numbers of branches and flowers, as well as more rapid flowering, fresh and dry flower weights, and physiological maturity. The highest number of female flowers was recorded from cuttings, irrespective of the cutting technique. Additionally, cannabinoid concentrations in Cannabis sativa oil were influenced by the propagation techniques. In the first growing season, herbaceous shoot cutting with 50% LA trimming had the highest CBD, while in the second growing season, the sexually propagated treatment had the highest CBD concentration. Additionally, herbaceous shoot cutting without LA trimming recorded the highest Δ9-THC concentration, followed by the treatment with 50% LA trimming during the first growing season. These findings indicate that asexual propagation through cuttings is a suitable propagation choice for flower production for pharmaceutical purposes, as female-only plants can be selected. However, sexual propagation should be used for fibre production.

1. Introduction

Cannabis sativa is a highly flexible plant, whose biomass production, secondary metabolite allocation (cannabinoids, terpenes, and other volatiles), and post-harvest essential oil quality are tightly controlled by both the genotype and propagation pathway used to produce the crop [1]. It produces considerable amounts of phytocannabinoids (a class of secondary metabolites), namely, cannabidiol (CBD) and tetrahydrocannabinol (THC), that are utilized in pharmaceutical applications [2,3]. Consequently, various laws have been established to support the policy of using cannabis species for pharmaceutical purposes, following established regulations, with a commitment to prioritizing the medicinal cannabis industry as a strategic path for the country’s economic development [4]. Phytocannabinoids are primarily concentrated in the essential oils extracted from the flowers of unfertilized female plants [5,6]. Conversely, fertilized female plants exhibit a decreased concentration and quality of phytocannabinoids, making male flowers less desirable within plantations [7]. Therefore, optimizing propagation methods tailored to specific end goals is essential for maximizing the crop growth and yield performance.
Cannabis is primarily propagated sexually from seed, which can lead to significant segregation due to genetic recombination [8]. This makes it challenging to maintain homogeneity in plants [6,7]. Its dioecious nature, heterozygosity, and inbreeding depression complicates seed production [9]. Consequently, the natural regeneration of this plant using seeds often leads to a high number of male flowering plants, which are undesirable for oil production, hindering the growth of the green economy [10].
The production of male populations leads to a decline in crop quality, particularly in locally produced cannabis landraces. This emphasizes the urgent need for alternative propagation methods, such as asexual propagation, to ensure superior produce for genetic conservation and pharmaceutical purposes. Asexual propagation through various cutting techniques is widely regarded as a crucial method for the rapid multiplication of true-to-type genotypes with high-quality genetic material [10]. It involves reproducing plant material from vegetative organs, ensuring that the progeny shares the exact characteristics of the parent plant in terms of their genotype and health status [11]. Propagation by cuttings ensures genetic uniformity and the production of female plants, making it a reliable and cost-effective method for cannabinoid production [12]. The literature suggests that plants produced through cuttings exhibit rapid development compared with those grown from seeds. This is because these cuttings consist of mature tissue, allowing them to bypass the seed germination phase [13]. Additionally, a single parent plant can produce many new plants, which is economically viable for nurseries and producers [14]. However, vegetative propagation through cuttings is often associated with production inefficiencies and a lack of scalability [15]. Thus, understanding how different propagation techniques affect the growth and development of the Cannabis sativa is critical for optimizing cultivation practices for commercial exploration, particularly in the Eastern Cape Province, which is known to harbour genetically diverse cannabis landraces that are locally adapted to the area’s ecological conditions [16]. This study seeks to identify the alternative and optimal propagation methods for the growth, development, yield, and oil quality of Cannabis sativa in the Eastern Cape Province.

2. Materials and Methods

2.1. Location of the Experiment

A pot experiment to determine the effect of the cutting (propagation) technique on the growth, development, oil yield, and quality of Cannabis sativa found in the Eastern Cape was conducted in a greenhouse under uncontrolled climatic conditions at DADI in Stutterheim, Eastern Cape Province (Figure 1). The study was conducted during the winter and summer growing seasons in 2024/25.

2.2. Planting Material and Treatments

This experiment used three propagation techniques (two asexual and one sexual), namely, rooted propagules of (i) herbaceous shoot cutting with 50% of the leaf area trimmed [expanded leaflet cut into half (HS50%LAT)], (ii) herbaceous shoot cutting with 100% leaf area [expanded leaflet not trimmed (AS100%LA)], and (iii) seedlings (sexual propagation) were used as treatments.

2.3. Experimental Design

The experiment was laid out in a Randomized Complete Design (RCD) (Figure 2). The three propagation techniques, i.e., two asexual cutting techniques and one sexual seedling technique, were used as treatments in this experiment and were replicated three times. Each treatment (propagation technique) consisted of ten (10) platelets per replicate, totaling thirty (30) per treatment. Therefore, the total sample size for the entire experiment was 90 plantlets. The treatment combination was as follows:
  • Herbaceous shoot with 50% leaf area trimmed (HS50%LAT);
  • Herbaceous shoot with 100% leaf area retained/not trimmed (HS100%LA);
  • Sexually produced seedlings (seedling).

2.4. Experimental Procedure

A 300 m2 (30 m × 10 m) greenhouse with a concrete surface and 20 L plastic pots was disinfected with spore-kill [(didecyldimethylammonium chloride 120 g/L), ICA International chemicals (Pty) LTD, Plankenburg Industrial, Stellenbosch, 7600, South Africa] before use in the experiment. The potting soil mixture, which comprised 2 parts Hygro-mix (Hygro-tech (Pty) LTD, Pretoria, South Africa) and 1 part mulch (commercial potting soils, Amalinda nursery, East London, South Africa), was prepared and used for experimental planting. Before planting, the mixture was analyzed for the basal nutrient composition at Dohne Analytical Laboratory (Table 1). Furthermore, 20 L pots were filled with prepared potting soil, following the planting principles in [17]. Uniform and healthy 20 cm tall seedlings and/or propagules were selected and transplanted into pots at one plant per pot. After transplanting (AT), the pots were kept in a greenhouse, watered to pot capacity, and monitored daily until the harvest was performed at 12 weeks after transplanting. Agronomic practices were adhered to as and when required.

2.5. Data Collection

(i) 
Vegetative growth and development: Plant height (cm), the number of lateral branches on one plant, and stem girth were measured using a flexible but non-elastic measuring tape, manual counting, and a Venier calliper (cm), respectively. Data were taken at two-week intervals after transplanting (AT) until the crop was harvested 12 weeks after transplanting. A total of five (5) plants per treatment were selected and tagged to measure the aforementioned vegetative growth parameters.
(ii) 
Flower yield:
  • The number of weeks to first flowering was determined;
  • The number of flowers and the flower sex determination per plant were performed through a physical count;
  • The number of weeks taken by plants to reach 50% amber coloration was recorded.
(iii) 
At harvest, five plants were destructively sampled, with plant stems cut at the first node above ground level, and the following biological yield parameters were obtained using an Adam ACBplus-6000g balanced scale (LBB6001e, Adam equipment, UK): (a) fresh plant weight (g) and (b) the leaves and flowers were removed from stems with the aid of secateurs and their fresh weight was measured. Samples were then dried and cured in dark-cool room conditions for 14 days, following the standards recommended by Jin et al. [18], and their dry weights were determined.
(iv) 
Oil yield and chemical composition: The oil yield and chemical composition were determined using gas chromatography (GC-MS) (Thermo Scientific, TriPlus RSH Smart, Switzerland) at Dohne Analytical Laboratory.
  • The oil yield was determined using an analytical balance (Mettler Toledo, MS104TS, Greifensee, Switzerland), which determined their mass with an accuracy of 0.001 g using the following formula (Demirel et al. [19]):
    O i l   y i e l d ( % ) = M a s s   o f   o i l M a s s   o f   a   s a m p l e × 100
  • The oil chemical composition was determined following the protocol by [20,21,22]. For instance, 310 grams (g) of dried flower was used for oil extraction through a hydro-distillation method using a Clevenger apparatus (WITEG, India). The vapour–oil mixture was passed through the condenser, where, after being condensed, was collected into a flask, and the oil was separated from water through a process known as decantation. The distillation period was 3 h, and the temperature of the heating mantle was 100 °C. Then, 10 µL of the sample was pipetted and diluted with 1 ml of ethanol. The sample was injected into the injector port. Gas chromatography–mass spectrometry (GC-MS) on a Thermo Fischer ISQ TM7610 single quadrupole with a fused silica polar capillary column (30 m × 0.25 mm × 0.25 µm film thickness) was used to analyze the essential oil. The oven temperature programming was from 50 to 250 °C and kept for 10 min at a rate of 2 °C per minute. The injector temperature of 250 °C and transfer line temperature of 280 °C were used. The mass spectrometer’s ion source and the analyzer were maintained at 280 °C and 100 °C, respectively. The mass spectrometer was conducted under full scan mode in electron impact ionisation (EI) positive mode, and the data was gathered from 40 to 600 m/z. The carrier gas was hydrogen at a rate of 1.2 mL/min. The chemical components were identified by comparing their relative retention times and mass spectra with data from the NIST library.

2.6. Data Analysis

The data was subjected to R-Studio statistical software version 4.2.2, United States (US), for analysis of variance (ANOVA). The comparison of means was performed using Fisher’s Least Significant Difference (LSD) (0.05), and the values were calculated at the p = 0.05 confidence level.

3. Results

3.1. Influence of Propagation Techniques on the Growth and Development of Cannabis sativa

The findings of the results, as highlighted in Table 2, showed that the propagation techniques significantly influenced the growth and development of Cannabis sativa in all the measured parameters, namely, plant height, number of branches per plant, stem girth, and number of weeks to flower amber coloration. Meanwhile, the results show that the number of weeks to first flowering was not significantly different. The tallest plants were obtained in plants produced from seedlings (sexual/seed), and the shortest were produced from propagules from the HS50%LAT treatment (73.3 cm) and were significantly different, whilst plants of HS100%LA (88.2 cm) were slightly different from both the seedlings and HS50%LAT. A similar trend was observed in the stem girth, where the highest and most significant number of branches was recorded in the HS100%LA treatment, with a total of 64 branches. This was followed by the HS50%LAT treatment, which produced 58 branches. In contrast, the plants grown from seedlings, which were produced sexually, exhibited the lowest branch count at 49. A similar trend was observed in the number of weeks until amber coloration. Although the differences were not statistically significant, plants treated with HS100%LA displayed the shortest duration to first flowering, taking only 4 weeks. This was followed by the HS50%LAT treatment, which required 5 weeks, while the seedlings took the longest at 9 weeks. A comparable trend was noted in season 2 (S2) throughout the experiment; however, the variations in weeks to first flowering and amber coloration were not statistically significant and were considered to be at par. Overall, the Cannabis sativa, when propagated from sexually produced seedlings, produced vigorous, tall plants that took longer to mature, whereas while not tall in terms of growth, the HS100%LA plants demonstrated the ability for rapid development and maturity, followed by HS50LAT.

3.2. Determination of Plant Gender as Influenced by Propagation Techniques Used in Cannabis sativa Production

The study demonstrated that the propagation techniques had a significant impact on the gender of Cannabis sativa plants in both seasons 1 and 2, respectively (Figure 3). In season 1, the plants produced through HS50%LAT (73.3%) recorded a significantly higher percentage (%) of female flowering plants, followed by HS100%LA (63.7%). However, sexually propagated seedling planting attained the highest male (50%) percentage and was insignificantly at par with the female (50%) flowering plants. In season 2, the HS100%LA treatment produced the highest percentage of female Cannabis sativa plants, with a significant rate of 83.3%. This was followed by the HS50%LAT treatment, but at a lower rate. In contrast, the seed treatment yielded the least, with only 33.3% of the plants being female. However, seed-produced plants attained the highest male flowering rate throughout the experiment.

3.3. Effect of Propagation Techniques on Biological Yield and Yield of Cannabis sativa

The findings in Table 3 indicate that the propagation techniques significantly influenced the biological yield and yield parameters measured in season 1, namely, the fresh plant weight (g), number of flowers per plant, fresh flower and leaf weights, and dry flower and leaf weights. The same occurred in season 2, except for the fresh plant and dry leaf weights, which were not significantly different, as highlighted in Table 4. The highest and significant fresh plant weight was found in the seedlings (236.1 g) and was significantly different from both HS100%LA and HS50%LAT, which were the same. The plants produced through the HS100%LA treatment demonstrated a significantly higher yield, producing the greatest number of flowers (202), as well as the highest fresh (99.5 g) and dry flower weights (45.3 g). This was followed by the HS50%LAT treatment, which yielded 163.5 flowers, with corresponding fresh and dry weights of 75.9 g and 25.8 g, respectively. In season 2, an insignificant difference was observed in the fresh plant weight, with seedlings recorded as the highest. Regarding the number of flowers, HS100%LA plants recorded the highest and were significantly different from sexually produced plants, followed by HS50%LAT, which was slightly significant relative to both the HS100%LA and sexually propagated plants. The fresh and dry flower weights were higher in HS100% LA, followed by HS50% LA, and the lowest were attained in seedlings, which were significantly different from both HS100% and HS50% LA, which were insignificantly the same. Sexually produced seedlings recorded the highest fresh and dry leaf weights, while the least was attained in HS50%LAT, which was significantly different from all the others.

3.4. Influence of Propagation Techniques on Oil Yield of Cannabis sativa

The results presented in Figure 4 indicate that plants propagated through the HS100%LA method exhibited the highest oil content during the winter cropping season, with a measured value of 0.6 g. This was followed by the HS50%LAT treatment, which yielded an oil content of 0.5 g. In contrast, the lowest oil content was observed in the seed-produced plants, which recorded only 0.34 g. A similar trend was observed in season two, where the HS100%LA treatment again yielded the highest oil content, consistent with previous findings. This was followed by HS50%LA, which also demonstrated a respectable oil yield. In contrast, the seed-produced plants recorded the lowest oil content at 0.3 g. Overall, the HS100%LA propagation technique produced a higher oil yield irrespective of the season.

3.5. Influence of Propagation Techniques on the Oil Quality of Cannabis sativa

Analysis of major cannabinoid concentrations revealed that propagation techniques influenced the oil quality of Cannabis sativa in both seasons. The oil analysis showed that in season one (S1), the oil extracted from cannabis flowers produced through HS50%LAT (28.0%) had the highest CBD compared with the HS100%LA and seed-derived plants, which had the same concentration (26.1%). In season two (S2), seed-derived plants (29.6%) had the highest CBD, followed by HS50%LAT (25.7%), and HS100%LA (24.5%) had the lowest concentration. A consistent trend was observed in the CBL, CBC, Δ8-THC, and Δ11-THC concentrations in both seasons. Except for Δ8-THC, which was higher in season 1, consistently higher concentrations of major constituents were obtained in season 2 compared with season 1, with seed-derived plants performing better, followed by HS50%LAT treatment. Vegetatively produced plants recorded the highest Δ9-THC (%) concentrations, with the highest attained in HS100%LA (15.8%), followed by HS50%LAT (11.9%) during season 1. Meanwhile, in season 2, the Δ9-THC (%) of HS50%LAT (11.7%) was higher, and HS100%LA had the lowest concentration. The findings further revealed that Δ11-THC followed a similar trend to Δ9-THC, where the 50% LAT treatments yielded higher concentrations irrespective of the season. The retention time (RT) was relatively stable across treatments, with 22.85 min in HS100%LA during season 1 and 24.40 min in the seedlings during season 2, with a slight increase in the seedlings.

4. Discussion

The results of this study indicate that Cannabis sativa was successfully propagated through vegetative cuttings. However, there were morphological differences between the seed-derived plants and those produced vegetatively [23]. Applying any of these asexual propagation methods is more important for increasing production [24]. The present study revealed that Cannabis sativa exhibited taller and more vigorous growth when propagated sexually from seeds compared with vegetative propagation. These findings align with those of Caplan [25], who reported the highest growth and vigour from cannabis propagated from seed rather than cuttings. Srikanth et al. [26] also noted that seedlings of Brassica species grow taller once the potential meristematic regions and root systems are established to support vegetative growth. Seed-cultivated plants develop a primary taproot system that offers better anchorage and access to soil water and nutrients [27]. This root system supports more substantial above-ground growth, resulting in taller and more vigorous plants [28]. Conversely, vegetatively propagated plants often develop fibrous root systems, which may not support as much vertical growth [29]. Additionally, vegetatively produced plants maintain uniformity but may lack the enhanced vigour sometimes seen in seed-grown plants [30].
The asexually propagated plants exhibited bushy growth with a high number of branches, as observed in HS100%LA, followed by HS50%LAT, compared with the seed-produced plants.
The results of this study align with the findings of Ioannidis et al. [31], who reported that vegetative cuttings tend to exhibit prolific lateral branch growth, leading to an increase in foliage compared with their parent plants. However, these results stand in contrast to the observations made by Jones et al. [32], who noted that vegetative propagation in cannabis often results in a high likelihood of apical dominance, which can limit branching and consequently reduce the overall foliage development. Formation of branches is attributable to the outbreak of axillary buds on the stem nodes [33]. In seed-grown plants, most sugars derived from photosynthesis are used to drive the growth of the shoot tip, whilst in asexually propagated plants, sugars rapidly accumulate in the axillary buds, triggering the cellular growth that results in branches [34]. The number of branches is determined by the number of nodes on the main stem [35]. The main stem, therefore, provides the basis of the development of all branches [30].
The vegetatively produced plants of HS100%LA, followed by HS50%LAT, demonstrated rapid flowering and physiological maturity compared with the seed-derived plants. Similar results were reported by Lazare et al. [36], where cannabis propagules with fully expanded leaves showed rapid maturation, which was attributable to low physiological stress, as well as the advanced growth stage during propagation. Cannabis cuttings with fully expanded leaf area may have the advantage during their early growth stages, as this enhances photosynthesis to provide sufficient carbohydrates and auxin for successful growth and development [27]. Vegetatively propagated plants result in faster establishment and earlier maturity since the propagated parts are already differentiated tissues [24]. Kurts et al. [37] concluded in their research that retip (leaf tip cut) propagules should be provided an extra week of vegetative growth to reach flowering to compensate for delays caused by initial physiological stress induced during propagation. However, while these plants may mature quicker, they do not always achieve the same ultimate height as seed-grown plants, which, despite a slower start, may continue growing for a more extended period [38].
The appearance of undifferentiated flower primordia in the stem at the nodes of the petiole and behind the stipule is the first indication of the flowering stage in the cannabis plant [39]. However, at this pre-floral stage, the cannabis flower genders cannot be identified [40]. Gender differences are primarily observable during the inflorescence stage, where males exhibit a branched inflorescence at the top of the plant with fewer leaves, while females possess their inflorescence at the apex of the stem [41]. Additionally, flowers on male cannabis plants typically emerge earlier than those on female plants, appearing approximately two weeks in advance [42]. Female cannabis plants serve as the primary source for medical cultivation, which is attributed to their elevated cannabinoid concentration found in the trichomes of the flowers [43]. The emerging results show that seed-derived plants exhibited the highest number of male plants, while the highest number of females was recorded from the vegetatively (cuttings) produced plants. Furthermore, a 50/50 trend was observed in seed-derived plants during the first season. These results concur with Trancoso et al. [44], who reported that seed propagation results in a roughly 1:1 ratio of male–female plants. Vegetative propagation guarantees genetic uniformity and increased production of female plants, an important attribute for dioecious essential oil hemp [45].
The study revealed that plants derived from sexual propagation demonstrated the highest fresh plant weight, as well as fresh and dry leaf weights, when compared with vegetatively produced plants, irrespective of the technique. This may be attributed to the fact that seed propagation supports the initial development of robust and well-established root systems, which are critical for effective nutrient and water absorption. These, in turn, promote enhanced plant growth and biomass accumulation [26]. Albrecht et al. [46] observed similar results, indicating that seed-derived citrus plants allocated a greater biomass to stems and leaves but had a poor fruit yield. Plants with a higher growth rate have an increased net photosynthesis that is converted and allocated to the total biomass of the plant [47].
Studies indicate that vegetatively propagated plants allocate more resources to reproductive structures, leading to enhanced flower production and a greater flower biomass [48,49,50,51]. This study demonstrated that cannabis plants propagated through cutting techniques often exhibit superior flowering characteristics compared with those grown from seeds, including a higher number of flowers and greater fresh and dry weights of flowers. These results concur with the findings of Stancheva et al. [52], who reported that Hyssopus officinale cuttings resulted in earlier flowering and higher flower biomass compared with the seed-grown counterparts. Since cuttings are genetically identical to the mother plant, they retain all the desirable traits and demonstrate consistent growth patterns, flowering times, and cannabinoid profiles [53,54].
In this study, vegetatively propagated plants subjected to 50% leaf area trimming (HS50% LAT) demonstrated higher CBD concentrations than their seed-derived counterparts during winter. For example, CBD synthesis is known to be influenced by photosynthetic activity, light exposure, and leaf physiology [55]. This may be attributed to the partial (50%) trimming of the leaf area that might have improved the light penetration into the plant canopy, reducing self-shading, and stimulating stress-responsive pathways, which are often linked to secondary metabolite production, including CBD. Vegetatively propagated plants (clonal cuttings) often exhibit enhanced uniformity, faster root establishment, and more robust secondary metabolite profiles due to their origin from mature, physiologically stable tissue [56]. Seed-derived plants consistently exhibited higher CBD concentrations than their vegetatively grown counterparts during the summer season. According to Morello et al. [57], longer photoperiods and a higher light intensity during the summer months enhance photosynthesis and secondary metabolite production, including cannabinoids. Accordingly, Arnao et al. [58] reported that plants grown from seed often develop more vigorous root systems and show stronger apical dominance, leading to a greater overall biomass and higher metabolic activity. These conditions aid robust CBD biosynthesis, especially when plants are not stressed by defoliation or restricted by clonal limitations.
The seed-produced plants, followed by those with 50% leaf area trimmed (HS50% LAT), revealed the highest concentrations of Cannabicyclol (CBL) and Cannabichromene (CBC), irrespective of the season. This might be because sexually propagated seedlings generally establish themselves more slowly than cuttings, allowing more time for CBC to accumulate. Over time this accumulation can convert to CBL through light-induced or oxidative processes. Subsequently, a diverse and open canopy may enhance light penetration and UV exposure, promoting CBC biosynthesis and CBL conversion [55].
In addition, seed-produced plants, followed by those with 50% leaf area trimmed (HS50% LAT), exhibited the highest concentrations of Δ8-tetrahydrocannabinol (Δ8-THC) and Δ11-tetrahydrocannabinol (Δ11-THC) across both the summer and winter seasons. Seed-derived cannabis plants are the most effective producers of Δ8-THC and Δ11-THC, likely due to their genetic and physiological advantages [39]. However, vegetatively propagated plants with 50% leaf area trimmed also perform well, likely due to the enhanced light exposure and stress signalling [59]. These findings emphasize the importance of propagation strategy and canopy management in maximizing the yield of THC isomers, regardless of the season.
The vegetatively propagated plants with no leaf area trimmed (HS100% LA) had the highest Δ9-THC concentrations, followed by the 50% leaf area trimmed (HS50% LAT) plants, whilst the sexually propagated plants consistently exhibited the lowest Δ9-THC levels across the seasons. Untrimmed plants maintain full foliage, which may create microenvironments, including higher humidity and lower UV exposure, that stabilize THC, reducing degradation into isomers like Δ8-THC or Δ11-THC [60]. Vegetative propagation produces genetically uniform plants with consistent metabolic programming, possibly favouring THC pathway dominance [61]. Moreover, trimming may have shifted some metabolic resources toward other cannabinoid branches, such as Δ8-THC or CBC, slightly lowering the Δ9-THC concentration compared with untrimmed cuttings [62]. Seedlings-derived (sexually propagated) plants with more open canopies might expose THC to light-driven degradation, converting it into Δ8-THC or Δ11-THC more readily [57,63].

5. Conclusions

The choice of propagation method significantly affects the morphological development and yield of Cannabis sativa. Plants produced asexually from cuttings showed the highest yield potential, regardless of the technique used, while those propagated from seed exhibited the greatest morphological growth and vigour. The study revealed that seed propagation results in many male plants with no economic value except for seed production. Seed propagation offers genetic diversity, whereas propagation by cuttings provides the genetic uniformity essential for pharmaceutical and commercial production. Seed-based propagation may be preferable for high CBD content, highlighting the sensitivity of CBD synthesis to leaf area and the importance of propagation technique to optimize cannabinoid yield. Additionally, the seed-derived cannabis plants had the highest concentrations of both CBL and CBC, demonstrating the benefits of genetic heterogeneity and natural growth patterns for minor cannabinoid production. However, 50% leaf area removal in the vegetatively propagated plants also proved positive, probably due to improved light exposure and stimulated metabolic activity. Remarkably, this trend was sustained across the seasons, strengthening the idea that propagation techniques and leaf area management, particularly partial trimming, have a greater influence on CBC and CBL concentrations than environmental factors alone. These findings suggest that strategic canopy management, combined with thoughtful propagation choices during seedling establishment stages, can significantly enhance minor cannabinoid yields in cannabis cultivation regardless of seasonal constraints. Unlike other cannabinoids, the Δ9-THC concentration was the highest in the vegetatively propagated plants with full leaf area (HS100% LA), demonstrating that maximum canopy preservation and genetic uniformity favour the accumulation of this main psychoactive compound. While partial trimming (HS50% LAT) still supports relatively high Δ9-THC production, seedlings underperform, possibly due to genetic variation, plant growth, and development dynamics that redirect cannabinoid synthesis toward alternative pathways. This insight is important for growers aiming for Δ9-THC-rich produce, as it highlights the value of vegetative propagation practices. Therefore, the study recommends asexual propagation through cuttings as the preferred method for flower cultivation, while seed should be used when plant height is desired in the fiber industry.

Author Contributions

A.D., Conceptualization, Writing—Initial Draft, Writing, Revisions, Data Curation, Investigation, and Visualization; I.M.E., Conceptualization, Writing, Review and Editing, and Supervision; T.T.S., Conceptualization, Writing, and Review and Editing; A.O.O., Data Curation and Reviewing, B.M., Conceptualization and Review and Editing; H.A.S., Reviewing and Editing. All authors have read and agreed to the published version of the manuscript.

Funding

This study received no external funding.

Institutional Review Board Statement

The study was conducted following the permit granted in terms of Section 22A(9)(a)(i) of the Medicines and Related Substances Act, 1965, to acquire, possess, and use Schedule 6 AND 7 substances for analytical and research purposes. Permit No: POS289/2025/2026. Granted by the South African Health Products Regulatory Authority (SAHPRA). The study was conducted according to ethical clearance as approved by the Ethics Committee of Walter Sisulu University (protocol number WSU/FNS-GREC/2024/03/11/G10 and date of approval 9 January 2024–16 January 2027).

Informed Consent Statement

Not applicable to the study.

Data Availability Statement

The original contributions presented in this study are included in the article/Supplementary Material. Further inquiries can be directed to the corresponding author(s).

Acknowledgments

The authors would like to express sincere gratitude to the Dohne Agricultural Research Development Institute and the Walter Sisulu University for providing the necessary resources and support during the study. Appreciation is also extended to all researchers and scientific technicians whose work contributed significantly to the foundation of this study. The authors extend their appreciation to Bulelwa Ngcangatha, Muzi Keswa, Thubela Tanki, Alina RamangoeleLungisa Joko and Olwethu Sindesi for their assistance with the GC-MS analysis, map construction, and statistical analysis.

Conflicts of Interest

The authors declare no conflicts of interest.

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Horticulturae 11 01428 g001
Figure 1. Map of Eastern Cape Province showing Döhne Agricultural Development Institute (DADI) where the experiment was conducted [source: Department of Agriculture (DOA, GIS)].
Figure 1. Map of Eastern Cape Province showing Döhne Agricultural Development Institute (DADI) where the experiment was conducted [source: Department of Agriculture (DOA, GIS)].
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Horticulturae 11 01428 g002
Figure 2. Schematic layout of the greenhouse experiment. HS50%LAT—herbaceous shoot with 50% leaf area trimmed, HS100%LA—herbaceous shoot with 100% leaf area retained/not trimmed, seedling—seed-produced seedlings.
Figure 2. Schematic layout of the greenhouse experiment. HS50%LAT—herbaceous shoot with 50% leaf area trimmed, HS100%LA—herbaceous shoot with 100% leaf area retained/not trimmed, seedling—seed-produced seedlings.
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Figure 3. Effect of propagation techniques on the percentage of male and female plants. S1: season 1, S2: season 2, HS50%LAT: herbaceous shoot with 50% leaf area trimmed, HS100%LA: herbaceous shoot with 100% leaf area retained/not trimmed, seedling: sexually produced plants from seed. Bars bearing the same letters indicate no significant differences (p = 0.05).
Figure 3. Effect of propagation techniques on the percentage of male and female plants. S1: season 1, S2: season 2, HS50%LAT: herbaceous shoot with 50% leaf area trimmed, HS100%LA: herbaceous shoot with 100% leaf area retained/not trimmed, seedling: sexually produced plants from seed. Bars bearing the same letters indicate no significant differences (p = 0.05).
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Figure 4. Effect of propagation techniques on the oil yield of Cannabis sativa. HS50%LAT: herbaceous shoot with 50% leaf area trimmed, HS100%LA: herbaceous shoot with 100% leaf area retained/not trimmed, seedling: sexually produced plants from seed. Bars bearing the same letters indicate no significant differences (p = 0.05).
Figure 4. Effect of propagation techniques on the oil yield of Cannabis sativa. HS50%LAT: herbaceous shoot with 50% leaf area trimmed, HS100%LA: herbaceous shoot with 100% leaf area retained/not trimmed, seedling: sexually produced plants from seed. Bars bearing the same letters indicate no significant differences (p = 0.05).
Horticulturae 11 01428 g004
Table 1. Nutrient composition of the prepared planting mixture before planting.
Table 1. Nutrient composition of the prepared planting mixture before planting.
Nutrient ElementValue
Phosphorus (P) (mg/L)514
Potassium (K) (mg/L)381
Calcium (Ca) (mg/L)2154
Magnesium (Mg) (mg/L)336
Zinc (Zn) (mg/L)17.8
pH (kcl)5.8
Table 2. Effects of propagation techniques on the growth and development of Cannabis sativa during seasons 1 and 2.
Table 2. Effects of propagation techniques on the growth and development of Cannabis sativa during seasons 1 and 2.
TreatmentPlant Height (cm)No. of Branches - 1Stem Girth (mm)No. of Weeks to FloweringNo. of Weeks to Flower Amber Coloration
S1
HS50%LAT73.3 (±16.4) 58 (±33.5) 11.8 (±1.53) 5 (±0.46) 8 (±0.57)
HS100%LA88.2 (±23.1) 64.1 (±29.1) 12.9 (± 1.81) 4 (±0.35) 6 (±0.69)
Seedling92.8 (±7.5) 49.3 (±14.6) 16.4 (±2.04) 9 (±0.81) 12 (±1.04)
Mean84.846.513.768.7
CV%26.649.113.89.48.5
p-Value0.05 *0.001 **0.05 *0.06 ns0.03 **
S2
HS50%LAT129.2 (±30.4) 53 (±8.1)10.9 (±5.03)8 (±0.57)9 (±1.4)
HS100%LA131.1 (±37.6) 71.3 (±17)11.4 (±4.9)8 (±0.57)9 (±1.4)
Seedling156.4 (±13.7) 58.2 (±8.6)12.7 (±5.4)10 (±1.2)12 (±2.23)
Mean138.960.811.78.610
CV%23.72415.29.79.4
p-Value0.05 *0.05 *0.001 **0.07 ns0.05 *
S1: season one, S2: season two, HS50%LAT: herbaceous shoot with 50% leaf area trimmed, HS100%LA: herbaceous shoot with 100% leaf area retained/not trimmed, seedling: seed-produced plants. ** Highly significantly different at p ≤ 0.05. * Significantly different at p ≤ 0.05. ns: not significantly different at the 5% probability level. p-Value: probability value. CV (%): coefficient of variation.
Table 3. Effect of propagation techniques on the biological yield of Cannabis sativa during seasons 1 and 2.
Table 3. Effect of propagation techniques on the biological yield of Cannabis sativa during seasons 1 and 2.
TreatmentMeans
Fresh Plant Weight (g)No of FlowersFresh Flower Weight (g)Fresh Leaf Weight (g)Dry Flower Weight (g)Dry Leaf Weight (g)
S1
HS50%LAT166.8 (±38)163.5 (±44.3)75.9 (±23)50.9 (±28)25.8 (±11)15.6 (±7.2)
HS100%LA167.5 (±37)202 (±81.2)99.5 (±22)53.8 (±24)45.3 (±7.4)18.3 (±7)
Seedling236.1 (±101)125 (±35)55.2 (±11)106.9 (±31)18.9 (±8)71.2 (±26)
Mean190.11637770.53035
Cv%35.3283337.54024.2
p-Value0.05 *0.01 **0.01 **0.05 *0.04 *0.05 *
S2
HS50%LAT415 (±92) 177.8 (±27)111.9 (±53)103 (±42)49 (±19)48 (±14.2)
HS100%LA469 (±57) 183.1 (±30.4)113.9 (±58.3)111 (±52)49.8 (±20)49.9 (±18)
Seedling476 (±91) 131.6 (±50)68.7 (±40.2)127.6 (±52)35.3 (±16.3)54.4 (±19)
Mean453.4164.298.2115.844.750.7
Cv%17.421.324.541.441.433
p-Value0.07 ns0.02 **0.05 *0.6 ns0.05 *0.57 ns
S1: season one, S2: season two, HS50%LAT: herbaceous shoot with 50% leaf area trimmed, HS100%LA: herbaceous shoot with 100% leaf area retained/not trimmed, seedling: sexually produced plants from seed. ** Highly significantly different at p ≤ 0.05. * Significantly different at p ≤ 0.05. ns: not significantly different at 5% probability level. p-Value: probability value. CV (%): coefficient of variation.
Table 4. Effect of propagation technique on the concentrations of major cannabinoids of Cannabis savita.
Table 4. Effect of propagation technique on the concentrations of major cannabinoids of Cannabis savita.
TreatmentCannabidiol (%)Cannabicyclol (%)Cannabichromene (%)Δ8-THC (%)Δ9-THC (%)Δ11-THC (%)
S1
HS50% LAT280.750.1343.311.98.4
 SH100% LA26.10.50.073515.86.26
 Seedling26.11.360.1853.87.67
S2
HS50% LAT25.71.20.154711.78.25
 HS100% LA24.30.70.0324.34.95.68
 Seedling29.620.2748.87.97.6
S1: season one, S2: season two, HS50%LAT: herbaceous shoot with 50% leaf area trimmed, HS100%LA: herbaceous shoot with 100% leaf area retained/not trimmed, seedling: sexually produced plants from seed. %: percentage, Δ8-THC: Delta 8-tetrahydrocannabinol, Δ9-THC: Delta 9-tetrahydrocannabinol, Δ11-THC: Delta 11-tetrahydrocannabinol.
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Dumani, A.; Silwana, T.T.; Egbichi, I.M.; Oyedeji, A.O.; Mpambani, B.; Seepe, H.A. Effect of Propagation Techniques on Growth, Development, Oil Yield, and Quality of Medicinal Cannabis (Cannabis sativa) Found in Lusikisiki, Eastern Cape, South Africa. Horticulturae 2025, 11, 1428. https://doi.org/10.3390/horticulturae11121428

AMA Style

Dumani A, Silwana TT, Egbichi IM, Oyedeji AO, Mpambani B, Seepe HA. Effect of Propagation Techniques on Growth, Development, Oil Yield, and Quality of Medicinal Cannabis (Cannabis sativa) Found in Lusikisiki, Eastern Cape, South Africa. Horticulturae. 2025; 11(12):1428. https://doi.org/10.3390/horticulturae11121428

Chicago/Turabian Style

Dumani, Azile, Tembakazi Theodora Silwana, Ifeanyi Moses Egbichi, Adebola Omowunmi Oyedeji, Babalwa Mpambani, and Hlabana Alfred Seepe. 2025. "Effect of Propagation Techniques on Growth, Development, Oil Yield, and Quality of Medicinal Cannabis (Cannabis sativa) Found in Lusikisiki, Eastern Cape, South Africa" Horticulturae 11, no. 12: 1428. https://doi.org/10.3390/horticulturae11121428

APA Style

Dumani, A., Silwana, T. T., Egbichi, I. M., Oyedeji, A. O., Mpambani, B., & Seepe, H. A. (2025). Effect of Propagation Techniques on Growth, Development, Oil Yield, and Quality of Medicinal Cannabis (Cannabis sativa) Found in Lusikisiki, Eastern Cape, South Africa. Horticulturae, 11(12), 1428. https://doi.org/10.3390/horticulturae11121428

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