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Medicinal Plant Research 2026, Vol.16, No.1 http://hortherbpublisher.com/index.php/mpr © 2026 HortHerb Publisher, registered at the publishing platform that is operated by Sophia Publishing Group, founded in British Columbia of Canada. All Rights Reserved. Publisher HortHerb Publisher Edited by Editorial Team of Medicinal Plant Research Email: edit@mpr.hortherbpublisher.com Website: http://hortherbpublisher.com/index.php/mpr Address: 11388 Stevenston Hwy, PO Box 96016, Richmond, V7A 5J5, British Columbia Canada Medicinal Plant Research (ISSN 1927-6508) is an open access, peer reviewed journal published online by HortHerb Publisher. The journal publishes all the latest and outstanding research articles, letters and reviews in all aspects of medicinal plant research, including plant growth and development, plant biology, plant nutrition, medicinal properties, phytochemical constituents, fitoterapia, pharmacognosy, essential oils, ethno- pharmacology agronomic management, and phytomedicine, as well as chemistry, pharmacology and use of medicinal plants and their derivatives. HortHerb Publisher is an international Open Access publisher specializing in horticulture, herbal sciences, and tea-related research registered at the publishing platform that is operated by Sophia Publishing Group (SPG), founded in British Columbia of Canada. All the articles published in Medicinal Plant Research are Open Access, and are distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. HortHerb Publisher uses CrossCheck service to identify academic plagiarism through the world’s leading plagiarism prevention tool, iParadigms, and to protect the original authors’ copyrights.
Medicinal Plant Research (online), 2026, Vol. 16, No.1 ISSN 1927-6508 http://hortherbpublisher.com/index.php/mpr © 2026 HortHerb Publisher, registered at the publishing platform that is operated by Sophia Publishing Group, founded in British Columbia of Canada. All Rights Reserved. Latest Content Bio Active Compounds and Antifungal Efficacy of Aqueous Extracts of Tridax procumbens Against Brown Rot of Amber Sweet Orange Fruits During Ambient Storage Oladele O.O., Oyedokun F.D., Adebomi D.M. Medicinal Plant Research, 2026, Vol. 16, No. 1, 1-10 Environmental Control and Standardized Production in Facility-Based Ganoderma lucidum Cultivation Xiaotong Wang Medicinal Plant Research, 2026, Vol. 16, No. 1, 11-30 Research on the Understory Ecological Cultivation Model of Tetrastigma hemsleyanum Weiduo Liu Medicinal Plant Research, 2026, Vol. 16, No. 1, 31-51 Hangbaiju Tea Beverages and Industrial Chain Extension Weiying Gao Medicinal Plant Research, 2026, Vol. 16, No. 1, 52-68 Effects of Cultivation Environment on the Yield of Fritillaria thunbergii and the Accumulation of Peimine Xiaoying Li, Jianhua Wang Medicinal Plant Research, 2026, Vol. 16, No. 1, 69-91
Medicinal Plant Research 2026, Vol.16, No.1, 1-10 http://hortherbpublisher.com/index.php/mpr 1 Research Report Open Access Bio Active Compounds and Antifungal Efficacy of Aqueous Extracts of Tridax procumbens Against Brown Rot of Amber Sweet Orange Fruits During Ambient Storage Oladele O.O. , Oyedokun F.D., Adebomi D.M. Department of Biology, School of Life Sciences, Federal University of Technology, PMB 704, Akure, Ondo State, Nigeria Corresponding author: kunle6634@gmail.com Medicinal Plant Research, 2026, Vol.16, No.1 doi: 10.5376/mpr.2026.16.0001 Received: 04 Feb., 2026 Accepted: 27 Feb., 2026 Published: 10 Mar., 2026 Copyright © 2026 Oladele et al., This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Preferred citation for this article: Oladele O.O., Oyedokun F.D., and Adebomi D.M., 2026, Bio active compounds and antifungal efficacy of aqueous extracts of Tridax procumbens against brown rot of amber sweet orange fruits during ambient storage, Medicinal Plant Research, 16(1): 1-10 (doi: 10.5376/mpr.2026.16.0001) Abstract This research evaluated the bio active compositions and efficacy of Tridax procumbens in controlling brown rot of orange fruits, stored at ambient temperature. Orange fruits infected with spore suspensions (6.50×104 cfu/ml) of brown rot pathogen (Lasiodiplodia sp) were dipped separately into varying concentrations (5, 10, 15 and 100%) of the aqueous extracts where they were assessed daily for disease severity. Each set up was in replicate of five orange fruits while the untreated fruits served as control. All the varying concentrations of the extracts were effective against Lasiodiplodia. However, 15 and 100 % extracts proved most effective against the brown rot pathogen, with none of the fruits showing any sign of infection by day 30 in storage when compared with the control fruits that had started showing rottenness since day 10. The number of bioactive compounds detected in the Tridax extract was 21. Consequently, the antifungal efficacy of Tridax procumbens could be attributed to these biochemical constituents. Hence, T. procumbens could be explored as natural, safe substitute in fruit preservation. Keywords Bioactive compounds; Disease severity; GC- MS; Lasiodiplodia; Preservatives Background Citrus fruits especially sweet orange is a good source of vitamin C, folic acid and fibres. In fact, sweet oranges are important exportable cash crops that serve as major source of foreign exchange to Nigeria prior to discovery of crude oil in 1951. In fact, it ranks as the most widely planted tree and earns substantial foreign exchange having produced 4.1 million tonnes in the year 2021 (FAOSTAT, 2023). The fruits are either eaten fresh or used for making canned orange juice. In Nigeria, the recommended varieties of sweet orange fruits include Hamlin, Valencia, Amber sweet, Agege Parson Brown, Umudike, Bende, Etir and Meran (Olaniyan et al., 2000). Nevertheless, the major cause of loss of orange fruits after harvest is pathological deteriorations. Thus, the potential foreign exchange earnings through the export of citrus fruits (sweet oranges) are under serious threats due to post-harvest infections, which could result in up to 40% yield loss. In fact, 40 to 50 percent of horticultural crops including fruits and vegetables are lost before they reach consumers while post-harvest losses in tropical fruits vary widely from 10 to 80 percent in both developed and developing countries. Damage so produced by diseases and pests is probably the major cause of loss in orange fruits. It was reported that Pencillium species, especially P. digitatum and P. italicum with Geotrichum citriaurantii are the major mould species causing post-harvest decay of sweet oranges (Eckert and Ogawa, 1985; Ohr and Eckert, 1985). However, Lasiodiplodia sp IMI50324 was reported as a major post-harvest pathogen of orange fruits in Akure, Nigeria, accounting for over 75 % disease incidence of the total rots in their survey study (Oladele and Aborisade, 2015). The fungus was identified by ITS Rdna sequence analysis using the FASTA algorithm with the fungus data base from European Molecular Biology Laboratory (EMBL) and the sequence showed 100% identity to numerous ITS sequences described from different Lasiodiplodia species or their Botryosphaeria teleomorphs. Even best matches with the fungus included sequences of Lasiodiplodia species reported in peer reviewed literature (Orlandelli et al., 2012).
Medicinal Plant Research 2026, Vol.16, No.1, 1-10 http://hortherbpublisher.com/index.php/mpr 2 Similarly, Lasiodiplodia was also reported as a frequent pathogen among sweet oranges in Brazil (Slippers and Wingfield, 2007), which is consistent with (Dantas et al., 2003) results for ‘Pera’ variety where the Lasiodiplodia rot was 53% of the total fruit rots while Wright and Harmon (2009) reported Lasiodiplodia rot as an important post-harvest disease in warm and humid growing regions such as Florida and Caribean. Twenty-four species of Lasiodiplodia are known to have been distinguished based on their DNA phylogeny together with their conidial morphology and morphology and size of their paraphyses (Burgess et al., 2006). Generally, all the known species of Lasiodiplodia are associated with various symptoms such as dieback, root rot, fruits rot and leaf spots among many others (Punithalingham, 1980). So, Lasiodiplodia sp IMI50324 is a common soil pathogen associated with woody hosts in the tropics and which causes brown rot in orange fruit (Oladele and Aborisade, 2015). Initial infection shows as light brown discolouration on any area of the fruit surface. As the decay develops, the lesion becomes more brown, firm and slippery (Ismail and Zhang, 2004). Currently, these pathogens are primarily controlled by application of fungicides either as dips, sprays, fumigants, treated wraps and box liners or in waxes and coatings. Citrus growers and sellers have routinely applied synthetic fungicides (chemicals) on their fruits for the management of post-harvest diseases. Numerous studies have been conducted to evaluate the effectiveness of various fungicides against common post-harvest pathogens affecting oranges, such as Penicillium digitatum and Penicillium italicum (Rosenberger et al., 2018), with none on Lasiodiplodia. Those studies provide important information for selecting appropriate fungicides and application protocols for combating post-harvest diseases in citrus fruits. Nonetheless, synthetic fungicides could result in health hazards during application process as well as residual accumulation in the fruits. Synthetic preservatives have raised concerns regarding their potential health risks and environmental impact, underscoring the need for innovative and eco-friendly solutions (Gupta et al., 2014). Besides, export markets are increasingly more sensitive to the use of chemicals for disease control coupled with the fact that most chemicals are expensive and inaccessible to local farmers who are the major bulk producers of this fruit in Nigeria. The escalating demand for safe and sustainable food preservation methods has prompted researchers to explore alternative solutions beyond conventional synthetic preservatives. Hence, the use of non-chemical ecofriendly means of control such as botanicals have emerged as viable alternatives. Besides, the rising awareness of health and environmental concerns associated with synthetic additives has led to a renewed interest in natural antimicrobial compounds derived from plant sources, thus making the utilization of plant-derived compounds as potential alternatives to synthetic additives to gain prominence, Therefore, the research investigated the antifungal efficacy of aqueous extract of Tridax procumbens against brown rot pathogen of orange fruits during ambient storage and its potential as a suitable alternative to synthetic preservatives. Phytochemical and bio active compositions of the aqueous extract was also investigated. 1 Materials and Methods 1.1 Source of fruits Mature, green healthy orange fruits were harvested from a commercial orchard in September 2023 from a citrus farm in Igbatoro, Akure North, Nigeria. Fruits of uniform size and colour were selected. Before treatment, the fruits were washed with clean water, disinfected for 10 min in 10 % sodium hypochlorite and allowed to air-dry at room temperature. 1.2 Preparation of spore suspension A ten-day old agar slant culture of Lasiodiplodia sp (IMI Number: 503248) on malt extract agar (MEA) was used to prepare spore suspension. Sterile water was poured into the slant and shaken vigorously to dislodge the spores from the vegetative hyphae. The wash water was collected in a sterilized beaker. One milliliter of the suspension was spread on an area of 1 cm2 and allowed to dry on a clean microscope slide before counting spores using the formula of Breed Direct Counting Technique (Ogundana, 1989) under the high dry ×40 objective microscope (Olympus).
Medicinal Plant Research 2026, Vol.16, No.1, 1-10 http://hortherbpublisher.com/index.php/mpr 3 1.3 Pathogenicity test Spore suspension of Lasiodiplodia was used to inoculate fresh fruits 1mm deep at the equator and incubated at 28±2℃ and 75% relative humidity inside sterilized desiccators. The disease symptoms were noted and re-isolation from infected fruit tissue was performed on fresh sterile MEA plate and its cultural characteristics were compared with the original isolate. 1.4 Preparation of extract of Tridax procumbens Detached Tridax procumbens leaves were air dried for 14 days. The dried leaves were pulverized with a blender to a smooth powdery form. About 28 g of the pulverized leaves were dissolved in 1 L of water, thoroughly shaken together and later filtered with a muslin cloth to obtain the crude /stock solution. Varying concentrations (5, 10, 15 and 100%) of the aqueous extract were then prepared from the stock / crude extract with appropriate volumes of water. 1.5 Inoculation and Treatment of orange fruits with Tridax procumbens extract The orange fruits were artificially infected with spores (6.50×104 cfu/ml) of Lasiodiplodia and later dipped separately into each concentration (5%, 10%, 15% and 100%) of the prepared extract for 5 min while benlated (fungicide) and untreated orange fruits served as positive and negative control respectively. Each set up consisted of 3 fruits. After treatment, the fruits were then placed inside a sterilized Petri dish and transferred into desiccators, and stored at 28± 2℃ and 75% relative humidity and assessed daily for disease severity 1.6 Assessment of rot severity Assessment of disease severity was done using the scale of (Arekemase and Oyeyiola, 2007) but with slight modification where 1=disease free, 2=slight rot /decay up to 10% of the fruit, 3=moderate rot /decay up to 25% of the fruit, and 4=severe rots/decay ≥35% of the fruit surface. Rot/decay was recognized by light brown discolouration on the fruit or by appearance of mycelium on the fruit surface. 1.7 Phytochemical screening of the aqueous extract of Tridax procumbens The phytochemical screening of the aqueous extract was done according to the method described by (Trease and Evans, 2004). The phytochemicals screened for were tannin, saponin, phlobatinnin, flavoniod, alkaloid and cardiac glycosides. 1.8 Gas chromatography-mass spectrophotometry (GC-MS) of Tridax procumbens extract The Tridax extract was subjected to chromatographic analysis using a Varian 3800/4000 gas chromatograph mass spectrometer equipped with an Agilent Technologies and a BP5 (30 m×0.25 mm×0.25 microns) capillary column. Nitrogen was used as a gas carrier. 1.0 µL volumes of the extract were injected using a splitless mode at an injector temperature of 270 ℃. The oven temperature was ramped from 80 to 200 ℃ (1 min hold) at a rate of 5 ℃/min. The oven temperature was held at 280 ℃ for 6 min following each analysis. The total run time for each sample was approximately 45 min. The GC-MS interface temperature was set to 280 ℃. The peaks of the organic compounds in the samples were identified in Wiley’s NIST 08 Mass Spectral Library and expressed in terms of its balance and retention time. 1.9 Statistical analysis The data obtained for the disease severity were subjected to analysis of variance and where significant, the means were compared at 5% level of probability using Tukey’s Test (SPSS version 20). 2 Results 2.1 Effects of different concentrations of Tridax procumbens extracts on disease severity in orange fruits pre- inoculated with Lasiodiplodia sp and stored at 28±2 ºC and 75 % relative humidity All treated and control fruits recorded mean disease severity values of 1.00±0.00 by day 5 of storage (Figure 1), indicating that they were all disease free. As storage duration progressed till day 10, except the control fruits that had shown slight rottenness as evident by their mean disease severity values of 2.00±0.52, all treated fruits were still disease free (healthy), though with mean disease severity values that were not significantly different (p≥0.05)
Medicinal Plant Research 2026, Vol.16, No.1, 1-10 http://hortherbpublisher.com/index.php/mpr 4 (Figure 2). For instance, fruits treated with both 15 and 100 % extracts of T. procumbens still maintained 1.00±0.00 as their disease severity values while fruits treated with 5, 10 % extracts of T. procumbens and fungicide had their severity values increase non-significantly (p ≥ 0.05) to 1.67±0.44, 1.33±0.38 and 1.33±0.38 respectively (Figure 2). Figure 1 Effect of different concentrations of Tridax procumbens on disease severity in orange fruits pre-inoculated with Lasiodiplodia sp and stored for 5 days Figure 2 Effect of different concentrations of Tridax procumbens on disease severity in orange fruits pre-inoculated with Lasiodiplodia sp and stored for 10 days Meanwhile by day 15 in storage, only fruits treated with 15 and 100% T. procumbens extracts maintained their diseased free status (1.00±0.00) when compared with control and other treated fruits. In fact, the severity values of control fruits had increased to 3.00±0.33 (moderate rottenness) while all the other treated fruits recorded same severity values (2.00±0.25) and had started showing slight rottenness (Figure 3). By day 20 in storage, only fruits treated with 15 and 100% T. procumbens extracts still maintained their diseased free status (1.00±0.00) when compared with control and other treated fruits (Figure 4). Unfortunately, the control fruits had become completely rotten (4.00±0.52) but fruits treated with 5 and 10% extracts of T. procumbens and fungicide still showed slight rottenness as evident by their respective severity values of 2.67±0.44, 2.50±0.50 and 3.00±0.33 (Figure 4).
Medicinal Plant Research 2026, Vol.16, No.1, 1-10 http://hortherbpublisher.com/index.php/mpr 5 Figure 3 Effect of different concentrations of Tridax procumbens on disease severity in orange fruits pre-inoculated with Lasiodiplodia sp and stored for 15 days Figure 4 Effect of different concentrations of Tridax procumbens on disease severity in orange fruits pre-inoculated with Lasiodiplodia sp and stored for 20 days Again by day 25 in storage, except fruits treated with 15 and 100% T. procumbens extracts that were still diseased free and control fruits that had become completely rotten, the severity values of all the other treated fruits with fungicide, 5 and 10% T. procumbens extracts had increased significantly to 3.67±0.67, 3.33±0.37 and 3.00±0.33 respectively, indicating moderate rottenness (Figure 5). The same trend of results was observed for all the fruits (control and treated) as the storage durations became further extended to day 30 (Figure 6). Figure 5 Effect of different concentrations of Tridax procumbens on disease severity in orange fruits pre-inoculated with Lasiodiplodia sp and stored for 25 days
Medicinal Plant Research 2026, Vol.16, No.1, 1-10 http://hortherbpublisher.com/index.php/mpr 6 Figure 6 Effect of different concentrations of Tridax procumbens on disease severity in orange fruits pre-inoculated with Lasiodiplodia sp and stored for 30 days 2.1 Phytochemical compositions of the aqueous extract of Tridax procumbens Tannins, alkaloids, cardiac glycosides, saponins and flavonoids were all detected in the Tridax extracts (Table 1). Table 1 Phytochemical constituents in Tridax procumbens Phytochemicals Aqueous extract Tannins + Alkaloids + Cardiac glycosides + Saponins + Flavonoids + 2.2 Bioactive compounds detected in aqueous Tridax procumbens extracts Table 2 shows the bioactive compounds with their peak area (%) detected in Tridax extract included 2,3-Butanediol (10.05), Hexanoic acid (11.77), Benzaldehyde,4-ethyl-(12.92), 4H-Pyran-4-one, 2,3 dihdro-3,5-dihydroxyl-6-methy-(6.89), Ethanone,1-(2-hydroxy-5-methylphenyl) (2.58), Methyl 4-(hydroxymethyl) benzoate (8.04), p-Menth-3-en-9-ol (3.73), Eugenol (1.44), Acetamide, N-tetrahydrofurfuryl-2-methoxy-(4.30), n-Hexadecanoic acid (12.64), 9,12-Octadecadienoic acid methy ester (1.03), Oleic acid (10.33), Octadecanoic acid (6.32) while the chromatograph of the aqueous extract of Tridax procumbens using GC-MS is shown in Figure 7. Table 2 Bioactive compounds detected in aqueous Tridax procumbens S/N Retention Time (min) Compound detected Chemical symbol Peak area (%) 1 8.00 2,3-Butanediol C4H10O2 10.05 2 9.46 Hexanoic acid C6H12O2 11.77 3 15.00 Benzaldehyde, 4-ethyl- C9H10O 12.92 4 16.62 4H-Pyran-4-one, 2,3-dihydro-3,5- dihydroxy-6-methyl- C6H8O4 6.89 5 21.00 Ethanone, 1-(2-hydroxy-5-methylphenyl)- C9H10O2 2.58 6 22.00 Methyl 4-(hydroxymethyl) Benzoate C9H10O3 8.04 7 22.50 p-Menth-3-en-9-ol C10H18O 3.73 8 24.51 Eugenol C10H12O2 1.44 9 26.00 Acetamide, N-tetrahydro furfuryl-2-methoxy- C8H15NO3 4.30 10 27.00 n-Hexadecanoic acid C16H32O2 12.64 11 29.57 9,12-Octadecadienoic acid methyl ester C19H34O2 1.03 12 30.84 Oleic acid C18H34O2 10.33 13 32.50 Octadecanoic acid C18H36O2 6.32
Medicinal Plant Research 2026, Vol.16, No.1, 1-10 http://hortherbpublisher.com/index.php/mpr 7 Figure 7 Chromatogram of aqueous extract of Tridax procumbens 3 Discussion This study explored the efficacy of aqueous leaf extracts of Tridax procumbens in controlling brown rot of orange fruits during ambient storage. In fact, the study showed that all the orange fruits treated with all the varying concentrations (5, 10, 15 and 100%) of the Tridax extracts were disease free (healthy), showing no sign of any infection by day 10 in storage, whereas the control fruits had started showing slight rottenness. Even when the storage durations became extended till day 30 and the control fruits had become completely rotten, fruits treated with 15 and 100% T. procumbens extracts were still disease free, without any sign of infection. Even performed better than the fungicide used (mancozeb) because all mancozeb treated fruits were moderately rotten by day 30. Though Rosenberger et al. (2018) demonstrated the effectiveness of various fungicides in controlling post-harvest rots of stone fruits, this work has further confirmed the use of botanicals in extending the shelf life of fruits better than fungicide because the aqueous leaf extracts of Tridax procumbens particularly 15 and 100% extracts, best protects the orange fruits for 30 days with no sign of brown rot/infection. This observation is also consistent with the works of Efunwoye et al. (2024) who observed that all the Padma tomato fruits treated with Tridax procumbens in their investigation exhibited significantly lower disease incidences (p ≤ 0.05) than the control fruits throughout the storage period. This showed the efficacy of botanicals in controlling fruit pathogens as equally buttressed by Zakawa et al. (2018) who investigated the use of neem leaf extracts in the control of post-harvest fungal pathogens on mango fruits in Yola, Adamawa state, Nigeria emphasizing the potential of the extract as a biofungicide and Udomlak et al. (2008) who demonstrated the antifungal efficacy of clove and cinnamon oil in preventing post-harvest decay in grape fruits, showing the synergistic potential of both oils in combating post-harvest fungi of grape fruits in vitro. Equally, Oladele (2019) investigated the antifungal activity of garlic extracts against three post-harvest pathogens that affect fruits, demonstrating garlic's potential as a natural preservative. In recent years, researchers have investigated the antimicrobial and antioxidant properties of Tridax procumbens extracts in fruit preservation. No wonder, Narendhirakannan and Subramanian (2005) reported that Tridax procumbens has been traditionally employed in various medicines for its pharmacological properties and antimicrobial activities in food preservation. Similarly, it has been reported that T. procumbens has played a major role in food preservation due to its rich phytochemical compositions with potential antimicrobial properties (Gupta et al., 2019). Notably, tannins,
Medicinal Plant Research 2026, Vol.16, No.1, 1-10 http://hortherbpublisher.com/index.php/mpr 8 alkaloids, cardiac glycosides, saponins and flavonoids were all detected as phytochemical constituents in the Tridax extracts in this work. This is in consonance with the findings of Jisha et al. (2016) and Palou et al. (2001) who reported a number of active chemical constituents such as alkaloids, flavonoids, carotenoids, β-sitosterol, fumeric acid, myristic, palmitic, arachidic, linoleic acid and tannin from botanicals and all these phytochemical composition, makes the plant a natural bio pesticide with potential antimicrobial efficacy against a broad spectrum of microorganisms (Nair et al. 2015; Mishra et al., 2018). Each of these phytochemical constituents as enunciated by others authors, is connected with significant antimicrobial properties. For instance, alkaloids and flavonoids when present exhibit antimicrobial properties. These compounds inhibit the growth of spoilage-causing microorganisms, extending the shelf life of fruits (Das, 2014). These compounds possess antioxidant properties, which play a crucial role in fruit preservation by inhibiting oxidative processes and prevent the degradation of fruits by scavenging free radicals that contribute to spoilage (Nair et al., 2015). This is further buttressed by the reports of Vasudevan et al. (2017) that alkaloids and flavonoids act as natural antimicrobial agents, inhibiting the growth of bacteria and fungi that contribute to fruit spoilage. Also, tannins act as natural preservatives by forming complexes with proteins and enzymes, thereby inhibiting microbial growth and enzymatic browning (Okigbo et al., 2010). Even the GC - MS analysis of the Tridax extract revealed a lot of bioactive compounds and these natural compounds exert their effects through various mechanisms, making them effective against different types of microorganisms. For instance, n-Hexadecanoic acid identified in the extract is reported to have pesticide activities (Rajasekaran and Cary, 2014). Also, 9-octadecenoic acid methyl ester possesses anti-microbial properties while stearic acid were also found to exhibit antibacterial and antifungal activities (Gehan et al., 2009). Remarkably, the presence of bioactive compounds in Tridax procumbens may contribute to natural pest resistance, presenting opportunities for sustainable pest management in agriculture. The method typically involves the application of plant extracts onto the fruit surface or as part of coatings, creating a protective barrier against fungal colonization. Besides, their anti-microbial properties inhibit the growth and development of post-harvest pathogens, thereby preserving the quality of the orange fruits. 4 Conclusions The study demonstrates antifungal efficacy of the aqueous extracts of Tridax procumbens against Lasiodiplodia sp (IMI: 50324), the brown rot pathogen of amber sweet orange fruits. The findings reveal the effectiveness of the aqueous extract at all tested concentrations against the brown rot pathogen by keeping the infected orange fruits healthy (disease free) for the first 10 days of storage under ambient temperature when compared with the untreated fruits. Further investigations even show that 15 and 100% concentrations of the aqueous extract still maintain the healthy status (wholesomeness) of the orange fruits for 30 days without any sign of rottenness as against the untreated and fungicide treated fruits that had already become unwholesome and diseased. Thus, we adjudge 15 and 100% of the Tridax aqueous extracts the most effective among the tested concentrations. They show potential for further development as safe, ecofriendly and effective natural substitutes against fruit pathogens. Author’s Contributions Oladele O.O. carried out the conceptualization, methodology, investigation, supervision, formal analysis and writing of original manuscript while Oyedokun F.D. and Adebomi D.M. were involved in the methodology, investigation, resources and data analysis. All authors read and approved the final manuscript. Acknowledgements The authors thank Life Technologies, UK for PCR purification and sequencing reactions of the fungi isolate. References Arekemase M.O., and Oyeyiola G.P., 2007, Fungi associated with spoiled citrus fruits obtained from Ilorin, Centre Point Science (Edition), 4: 138-149. Burgess T.I., Barber P.A., Mohali S., Pegg G., and Wingfield M.J., 2006, Three new Lasiodiplodia spp. from the tropics, recognized based on DNA sequence comparisons and morphology, Mycologia, 98(3): 423-435. https://doi.org/10.1080/15572536.2006.11832677
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Medicinal Plant Research 2026, Vol.16, No.1, 11-30 http://hortherbpublisher.com/index.php/mpr 11 Feature Review Open Access Environmental Control and Standardized Production in Facility-Based Ganoderma lucidum Cultivation Xiaotong Wang Jinhua Shouxiangu Pharmaceutical Co. Ltd., Jinhua, 321200, Zhejiang, China Corresponding email: 378968737@qq.com Medicinal Plant Research, 2026, Vol.16, No.1 doi: 10.5376/mpr.2026.16.0002 Received: 15 Jan., 2026 Accepted: 01 Mar., 2026 Published: 16 Mar., 2026 Copyright © 2026 Wang, This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Preferred citation for this article: Wang X.Y., 2026, Environmental control and standardized production in facility-based Ganoderma lucidum cultivation, Medicinal Plant Research, 16(1): 11-30 (doi: 10.5376/mpr.2026.16.0002) Abstract This study explores the key technical pathways for environmental control and standardized production in facility-based Ganoderma lucidum cultivation. As market demand for G. lucidum products continues to expand, the limitations of traditional cultivation methods in precise environmental regulation, product quality consistency, and production standardization have become increasingly evident. This paper analyzes the specific requirements of G. lucidum growth for temperature, humidity, light, CO2 concentration, substrate, and a clean cultivation environment. It summarizes facility-based production models, including greenhouse cultivation, intelligent mushroom houses, multi-layer bag cultivation, and liquid fermentation, with emphasis on temperature and humidity control, ventilation and light regulation, clean-environment management, and pest and disease prevention. Meanwhile, the standardized production process of G. lucidum is reviewed in terms of strain selection, substrate standardization, cultivation bag preparation, sterilization and inoculation, fruiting management, harvesting and processing, and quality evaluation. The study suggests that the application of IoT monitoring, automated regulation, data analysis, and intelligent management technologies can improve the precision, standardization, and traceability of G. lucidum production. In the future, regionalized cultivation technology systems, breeding and promotion of superior varieties, and branding and product traceability systems should be further strengthened to promote the high-quality, green, and intelligent development of the G. lucidum industry. Keywords Ganoderma lucidum; Facility-based cultivation; Environmental control; Standardized production; Intelligent management 1 Introduction Ganoderma lucidum is one of the traditional valuable medicinal fungi in China. It was classified as a “superior-grade” medicinal material in Shennong’s Classic of Materia Medica and has long been recognized for its medicinal value in tonifying qi, calming the mind, and strengthening healthy qi. Modern studies have shown that G. lucidum is rich in bioactive compounds such as polysaccharides and triterpenoids, which exhibit various pharmacological activities, including immunomodulatory, antioxidant, antitumor, and metabolic regulatory effects. Therefore, its application value in food, nutritional supplements, pharmaceuticals, functional products, and wellness markets has continued to increase (Thakur et al., 2024). In recent years, with the rapid development of the health industry and the increasing health awareness of consumers, market demand for G. lucidum-related products has continued to expand. The stable and large-scale supply of safe, high-quality G. lucidum raw materials and derivative products has become an important foundation for supporting industrial development (Wu et al., 2024). However, traditional G. lucidum cultivation systems still have certain limitations in yield stability, quality consistency, resource-use efficiency, and contamination control. Some production practices still rely on experience-based management, which may lead to imprecise environmental regulation, large variations in product quality, frequent occurrence of pests and diseases, and inconsistent production standards. These problems make it difficult to meet modern market demand for high-quality, safe, and stable G. lucidum products (Ghafoor et al., 2024). Therefore, promoting technological upgrading and the construction of standardized production systems has become a practical requirement for the high-quality development of the G. lucidum industry. G. lucidum is a typical wood-decaying fungus, and its mycelial growth and fruiting body formation are highly sensitive to environmental conditions such as temperature, humidity, light, air circulation, CO2 concentration, and
Medicinal Plant Research 2026, Vol.16, No.1, 11-30 http://hortherbpublisher.com/index.php/mpr 12 nutrient supply. Under traditional open-field cultivation or simple greenhouse cultivation, G. lucidum production is easily affected by seasonal changes, climatic fluctuations, and natural disasters, often resulting in slow mycelial growth, abnormal fruiting body morphology, unstable yield, and fluctuations in active component contents. Meanwhile, the scarcity and instability of wild resources, high dependence on wood-based substrates, and obvious batch-to-batch differences in active components further highlight the need to establish efficient, controllable, and sustainable production systems. With the development of modern agricultural facility technologies and bioprocessing technologies, facility-based cultivation has gradually become an important direction for the transformation and upgrading of the G. lucidum industry. Indoor mushroom houses, environmentally controllable “mushroom factories,” liquid spawn systems, and submerged fermentation technologies can precisely regulate temperature, humidity, gas composition, light, and nutritional conditions, thereby improving production stability and quality consistency (Feng et al., 2024; Ghafoor et al., 2024; Liu et al., 2024). In addition, facility-based cultivation can reduce the risks associated with heavy metals, toxic residues, and competitive microorganisms by separating substrates from soil, while also supporting standardized operations that comply with pharmacopoeial requirements and good manufacturing practices (Wu et al., 2024). In facility-based G. lucidum cultivation, environmental control technology is a core factor affecting production performance and product quality. Temperature and humidity are directly related to mycelial expansion, primordium formation, and fruiting body development; ventilation affects changes in CO2 concentration and plays an important role in pileus formation, stipe elongation, and morphogenesis; light conditions are closely associated with primordium differentiation, color formation, and the accumulation of bioactive substances. Studies have shown that controlled environments can optimize fruiting body morphology, yield, and the accumulation of bioactive components such as polysaccharides and triterpenoids by regulating key factors such as light spectrum, temperature profiles, and oxygen supply. Improper environmental regulation can easily lead to contamination, excessive stipe elongation, malformed pilei, and insufficient accumulation of active components. Therefore, establishing a scientific, stable, and reproducible environmental control system is an important basis for improving the level of facility-based G. lucidum cultivation. Meanwhile, the application of low-cost IoT-based temperature and humidity sensing, automatic control, and real-time monitoring technologies has further enhanced environmental management during G. lucidum production, helping to achieve contaminant control, compliance with quality indicators, and standardized whole-process management (Nguyen et al., 2023; Liu et al., 2024). This study will explore the key technical pathways for environmental control and standardized production in facility-based G. lucidum cultivation. It will focus on the environmental regulation points, key production procedures, and quality control systems of facility-based G. lucidum cultivation; examine the application value of intelligent and digital technologies in G. lucidum production; and propose development pathways for standardized production in light of the current industrial context. The study aims to integrate the development needs of the G. lucidum industry with modern controllable cultivation technologies. By summarizing research progress in facility design, temperature, humidity, CO2, light quality, ventilation regulation, liquid spawn, and fermentation processes, it seeks to provide technical references for stable and high-quality G. lucidum production. This study will also examine how precise environmental regulation can support emerging quality standards, increase the yield of fruiting bodies and bioactive metabolites, reduce contamination and resource consumption, and thereby promote G. lucidum production toward precision, standardization, and intelligentization, enhance product quality and industrial competitiveness, and facilitate the sustainable and high-quality development of the G. lucidum industry. 2 Basic Characteristics of Facility-Based Ganoderma lucidum Cultivation 2.1 Specific environmental requirements for Ganoderma lucidum growth Ganoderma lucidum is a thermophilic wood-decaying fungus. Its mycelial growth, primordium differentiation, and fruiting body formation show distinct stage-specific requirements for temperature, humidity, light, CO2 concentration, pH, and substrate conditions. Studies have shown that G. lucidum mycelia generally grow well at 25℃~32℃, under near-neutral to slightly acidic pH and relatively high humidity, whereas fruiting body formation depends more on stable temperature and humidity, appropriate light, and favorable gas exchange
Medicinal Plant Research 2026, Vol.16, No.1, 11-30 http://hortherbpublisher.com/index.php/mpr 13 conditions (Al-Kaabi and Hussien, 2025). For example, Magday (2014) found that a wild Philippine strain of G. lucidum showed better mycelial growth at pH 6.0, 32℃, and under light conditions; Lengare et al. (2023) reported that vegetative growth performed well at 25℃, 95%-100% relative humidity, and a 12 h light/12 h dark cycle. Therefore, facility-based G. lucidum cultivation is not merely the control of a single environmental parameter, but requires integrated regulation according to different growth stages. During fruiting body formation, G. lucidum has stricter requirements for air humidity, light, and ventilation. Appropriate humidity promotes primordium formation and pileus expansion, whereas insufficient ventilation may lead to elevated CO2 concentration in the facility, resulting in elongated stipes, poor pileus development, or abnormal morphology. A Korean bed-cultivation study showed that 28℃~30℃, light intensity of 1 000~1 500 lux, and approximately 1% CO2 were suitable for fruiting body formation. Recent studies on light quality regulation further indicate that green light can promote stipe elongation, pileus expansion, fresh weight increase, and the contents of crude protein, polysaccharides, triterpenoids, and extracellular enzyme activity, whereas red light may inhibit fruiting body differentiation. Therefore, spectral selection and lighting regime design should be emphasized in factory cultivation (Liu et al., 2024). This suggests that light not only affects the appearance quality of G. lucidum, but is also closely related to the accumulation of active components. In addition to physical environmental factors, substrate chemical properties and facility cleanliness are also important factors affecting stable G. lucidum production. Continuous cultivation can alter organic matter, pH, salinity, and enzyme activities in soil or substrates, leading to changes in the metabolite composition of fruiting bodies (Wang et al., 2022). Meanwhile, competitive fungi such as Trichoderma can proliferate readily in G. lucidum cultivation environments and inhibit G. lucidum growth through volatile and non-volatile metabolites, causing contamination and yield loss (Lim et al., 2024). Therefore, facility-based G. lucidum production should integrate temperature and humidity regulation, light management, CO2 control, pH and salinity adjustment, substrate disinfection, and spatial sanitation management to establish a stable and reproducible environmental control system. 2.2 Main models of facility-based Ganoderma lucidum cultivation With the development of modern agricultural facility technologies, G. lucidum production has gradually shifted from traditional open-field cultivation and simple greenhouse cultivation toward facility-based, controllable, and standardized models. At present, greenhouses and indoor mushroom houses are common solid-state cultivation models for fruiting body production. They usually use sawdust, agricultural residues, or logs as substrates, and are equipped with functional areas such as inoculation rooms, incubation rooms, sterilization rooms, fruiting rooms, packaging rooms, and waste disposal areas (Thakur et al., 2024; Akçay et al., 2025). Standardized bed cultivation generally uses formulated sawdust-based substrates and promotes fruiting body formation under controlled temperature, light, and CO2 conditions. Compared with traditional open-field cultivation, greenhouse and mushroom-house cultivation can reduce the influence of external climate fluctuations and improve production stability and year-round supply capacity. In solid-state facility cultivation, bag cultivation and multi-layer rack cultivation are important forms of current large-scale production. Bag cultivation uses lignocellulosic wastes such as sawdust, rice straw, wheat straw, and hazelnut shells as main raw materials. By optimizing substrate formulations, it can shorten mycelial colonization time and improve yield and the number of fruiting flushes (Akçay et al., 2025). Multi-layer rack cultivation improves space utilization by vertically arranging cultivation bags, facilitating unified sterilization, inoculation, incubation, and fruiting management. Some regions have also developed permanent greenhouse sand-bed cultivation models, in which cultivation substrates are separated from soil using plastic membranes and combined with sprinkler and shading systems. This can reduce the risks of heavy metals, pesticide residues, and competitive microbial contamination, and increase yield by approximately 30% compared with traditional open-field methods. This type of model combines environmental controllability with quality and safety advantages, making it suitable for standardized medicinal G. lucidum production.
Medicinal Plant Research 2026, Vol.16, No.1, 11-30 http://hortherbpublisher.com/index.php/mpr 14 In addition to solid-state fruiting body production, liquid cultivation and bioreactor systems have gradually become important components of facility-based G. lucidum production. Liquid facility-based models mainly target mycelial biomass, extracellular polysaccharides (EPS), triterpenoids, and other metabolites, and generally use shake flasks, stirred-tank reactors, or specially designed bioreactors for closed cultivation (Alsaheb et al., 2020). For example, new reactors such as the Air-L-Shaped Bioreactor (ALSB) can reduce mycelial adhesion to the reactor wall and clumping, thereby improving mass transfer efficiency and process economics (Supramani et al., 2023). In addition, two-stage liquid static cultivation and bag-type static bioreactors can be used for efficient accumulation of triterpenoids, indicating that facility-based G. lucidum cultivation has expanded from simple fruiting body production to targeted production of mycelia and functional metabolites. Overall, facility-based G. lucidum cultivation is forming a diversified system ranging from greenhouse cultivation, intelligent mushroom houses, and multi-layer bag cultivation to liquid fermentation and bioreactor production. 2.3 Development characteristics of large-scale Ganoderma lucidum production In recent years, the G. lucidum industry has increasingly shown trends toward scaling-up, intensification, engineering, and standardization. Traditional solid-state cultivation based on logs or beds has a relatively long production cycle and high labor intensity, and is vulnerable to contamination, continuous cropping obstacles, and environmental fluctuations. These limitations make it difficult to meet global market demand for stable supply and quality consistency (Araque et al., 2020; Wu et al., 2024). As G. lucidum is increasingly used in dietary supplements, pharmaceuticals, functional foods, and health products, large-scale production requires not only higher yield, but also unified standards for active components, safety, and batch consistency. Through unified strain supply, standardized substrate preparation, centralized cultivation management, and batch-based production processes, production costs per unit can be effectively reduced, production efficiency can be improved, and product quality stability can be enhanced. During large-scale development, facility-based cultivation and process engineering have become important technical supports. Engineering simulation studies have shown that tools such as SuperPro Designer can be used to simulate industrial-scale submerged cultivation of G. lucidum, predict equipment requirements, energy consumption, and operating costs, and evaluate the economic effects of bioreactor scale-up. For example, increasing bioreactor volume from 2 m3 to 20 m3 can significantly reduce the unit production cost of extracellular polysaccharides and other products, demonstrating the importance of economies of scale and process optimization (Araque et al., 2020). Semi-industrial and pilot-scale studies have also shown that controlled pH, optimized carbon and nitrogen sources, and appropriate reactor configurations can significantly improve the yields of extracellular polysaccharides and other metabolites (Alsaheb et al., 2020; Supramani et al., 2023). This indicates that large-scale G. lucidum production is no longer limited to traditional cultivation experience, but is gradually moving toward process design, parameter optimization, and engineering scale-up. Standardized management is an important approach to enhancing the competitiveness of the large-scale G. lucidum industry. At present, G. lucidum production has shifted from merely pursuing yield to placing greater emphasis on quality, safety, traceability, and market standardization. Relevant reviews have indicated that industrial standardization of G. lucidum is promoting the standardization of raw material sources, processing procedures, quality control, and product circulation, while emphasizing the importance of active component content, hygiene and safety, and production consistency in industrial development. Meanwhile, facility-based cultivation, non-soil-contact systems, and clean-environment management can help reduce heavy metal contamination and the hazards of competitive fungi, thereby improving production reproducibility and quality stability (Lim et al., 2024; Wu et al., 2024). In the future, with the further application of environmental monitoring, intelligent ventilation, automatic spraying, data analysis platforms, and bioreactor technologies, large-scale G. lucidum production will place greater emphasis on precise regulation, green production, brand-oriented operation, and alignment with international quality standards, thereby promoting the industry toward high-quality development.
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