First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm (Elaeis guineensis Jacq.) in Ghana

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Osekre" }, { "@type": "Person", "name": "Frank K. Ackah" } ], "publisher": { "@type": "Organization", "name": "F1000Research", "logo": { "@type": "ImageObject", "url": "https://f1000research.com/img/AMP/F1000Research_image.png", "height": 480, "width": 60 } }, "image": { "@type": "ImageObject", "url": "https://f1000research.com/img/AMP/F1000Research_image.png", "height": 1200, "width": 150 }, "description": " Backgrounds Oil palm (Elaeis guineensis Jacq.), is the most significant and highest-yielding crop among oil-producing crops worldwide. In 2020/2022, Basal stem rot (BSR) disease was observed in six oil palm growing Districts in Ghana. Methods Field study and laboratory analysis were conducted. A random sampling technique was used to select five plantation blocks from each District. Single-point disease assessments were done using Standard Operating Procedure (SOP) with a severity scale of 0-4. Molecular assays were performed on each sample using nucleic acid as a template. ITS and GanET sequence analysis were performed along with the formation of a phylogenetic tree using the FASTA algorithm with the Fungus database from EBI and NCBI GenBank. Koch’s postulate was followed to confirm the disease. Results The disease incidence was 11.3 % with the highest severity score of 4. BSR is characterised by stem decay large-perennial, woody brackets basidiocarps of average measurement of 2-65 cm in diameter on infected palms. Culture colonies were white, striated, undulating, woolly-cottony, and creamish pigment on the reverse depicting attributes of Ganoderma fungus. Molecular confirmation was done by combining ITS sequence of top matches of >97% to members of the genus Ganoderma, >98% and 99.3% identity to three sequences of Ganoderma sp. (HM138671; HM138670 and HM138672) generated from strains assigned to Ganoderma ryvardenii and compared with 132 published sequences of Ganoderma isolates. Conclusion This is the first report of Ganoderma ryvardenii causing BSR disease on oil palm in Ghana and possibly the second report in Africa. However, the pathogen was first reported to cause similar diseases in oil palm in Cameroon. " } { "@context": "http://schema.org", "@type": "BreadcrumbList", "itemListElement": [ { "@type": "ListItem", "position": "1", "item": { "@id": "https://f1000research.com/", "name": "Home" } }, { "@type": "ListItem", "position": "2", "item": { "@id": "https://f1000research.com/browse/articles", "name": "Browse" } }, { "@type": "ListItem", "position": "3", "item": { "@id": "https://f1000research.com/articles/14-413/v1", "name": "First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR)..." } } ] } Home Browse First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR)... ALL Metrics - Views Downloads Get PDF Get XML Cite How to cite this article Lekete-Lawson E, van der Puije GC, Osekre EA and Ackah FK. First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved, 2 approved with reservations] . F1000Research 2025, 14 :413 ( https://doi.org/10.12688/f1000research.161972.1 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. Close Copy Citation Details Export Export Citation Sciwheel EndNote Ref. Manager Bibtex ProCite Sente EXPORT Select a format first Track Share ▬ ✚ Research Article First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved, 2 approved with reservations] Emmanuellah Lekete-Lawson https://orcid.org/0000-0002-9478-8833 1-3 , Grace C. van der Puije 2 , Enoch A. Osekre 4 , Frank K. Ackah https://orcid.org/0000-0001-9188-7476 2 Emmanuellah Lekete-Lawson https://orcid.org/0000-0002-9478-8833 1-3 , Grace C. van der Puije 2 , Enoch A. Osekre 4 , Frank K. Ackah https://orcid.org/0000-0001-9188-7476 2 PUBLISHED 08 Apr 2025 Author details Author details 1 CSIR-Oil Palm Research Institute, Kade, Eastern Region, Ghana 2 Department of Crop Science, University of Cape Coast College of Agriculture and Natural Sciences, Cape Coast, Ghana 3 Department of Microbiology, Nutrition and Dietetics, Czech University of Life Sciences, Faculty of Agrobiology Food and Natural Resources, Prague, Czech Republic 4 Department of Crop and Soil Sciences, Kwame Nkrumah University of Science and Technology College of Agriculture and Natural Resources, Kumasi, Ghana Emmanuellah Lekete-Lawson Roles: Investigation, Methodology, Resources, Writing – Original Draft Preparation Grace C. van der Puije Roles: Supervision Enoch A. Osekre Roles: Supervision, Writing – Review & Editing Frank K. Ackah Roles: Supervision OPEN PEER REVIEW DETAILS REVIEWER STATUS This article is included in the Plant Science gateway. Abstract Backgrounds Oil palm ( Elaeis guineensis Jacq.), is the most significant and highest-yielding crop among oil-producing crops worldwide. In 2020/2022, Basal stem rot (BSR) disease was observed in six oil palm growing Districts in Ghana. Methods Field study and laboratory analysis were conducted. A random sampling technique was used to select five plantation blocks from each District. Single-point disease assessments were done using Standard Operating Procedure (SOP) with a severity scale of 0-4. Molecular assays were performed on each sample using nucleic acid as a template. ITS and GanET sequence analysis were performed along with the formation of a phylogenetic tree using the FASTA algorithm with the Fungus database from EBI and NCBI GenBank. Koch’s postulate was followed to confirm the disease. Results The disease incidence was 11.3 % with the highest severity score of 4. BSR is characterised by stem decay large-perennial, woody brackets basidiocarps of average measurement of 2-65 cm in diameter on infected palms. Culture colonies were white, striated, undulating, woolly-cottony, and creamish pigment on the reverse depicting attributes of Ganoderma fungus. Molecular confirmation was done by combining ITS sequence of top matches of >97% to members of the genus Ganoderma , >98% and 99.3% identity to three sequences of Ganoderma sp. (HM138671; HM138670 and HM138672) generated from strains assigned to Ganoderma ryvardenii and compared with 132 published sequences of Ganoderma isolates. Conclusion This is the first report of Ganoderma ryvardenii causing BSR disease on oil palm in Ghana and possibly the second report in Africa. However, the pathogen was first reported to cause similar diseases in oil palm in Cameroon. READ ALL READ LESS Keywords Ganoderma disease, Basal Stem Rot, oil palm, Ghana, phylogenetic Corresponding Author(s) Emmanuellah Lekete-Lawson ( [email protected] ) Close Corresponding author: Emmanuellah Lekete-Lawson Competing interests: No competing interests were disclosed. Grant information: The author(s) declared that no grants were involved in supporting this work. Copyright: © 2025 Lekete-Lawson E et al . This is an open access article 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. How to cite: Lekete-Lawson E, van der Puije GC, Osekre EA and Ackah FK. First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved, 2 approved with reservations] . F1000Research 2025, 14 :413 ( https://doi.org/10.12688/f1000research.161972.1 ) First published: 08 Apr 2025, 14 :413 ( https://doi.org/10.12688/f1000research.161972.1 ) Latest published: 07 Jul 2025, 14 :413 ( https://doi.org/10.12688/f1000research.161972.2 )  There is a newer version of this article available. Suppress this message for one day. 1. Introduction Oil palm ( Elaeis guineensis Jacq), is considered a worldwide pre-eminent and highest-yielding edible oil crop among other oil-producing crops and has since inspired many economically emergent nations including Malaysia, Indonesia, India, who engaged in oil palm production ( Ghulam, 2022 ). Although oil palm is known to have originated from Africa, only 73.8 million metric tons, representing 3%, was produced by the continent in 2021/22 ( Biney et al., 2024 ), with West Africa making 4% ( FAOSTAT, 2022 ). Ghana's oil palm production is about 0.5 million metric tons annually ( Biney et al., 2024 ; Sasu, 2023 ), representing a tiny portion of the global total despite being endowed with high-yielding varieties. Although Ghana can produce more oil palm, challenges including pests and diseases as well as climate variability prevent it from expanding ( Zutah et al., 2024 ). Nonetheless, by addressing these challenges, Ghana will improve its oil palm production and ensure less dependency on food imports. Among the diseases reported on oil palms in Ghana is Basal Stem Rot (BSR), which is suspected to be caused by the white rot fungus called Ganoderma . Ganoderma sp. is a basidiomycetes fungus and has been reported to be the primary cause of BSR disease. It is the most devastating and destructive disease of oil palm in tropical regions and causes substantial economic losses up to 43% biannually to oil palm producers ( Khoo & Chong, 2023 ). According to Karunarathna et al. (2024) , BSR disease can cause yield reductions of up to 80% in severely affected plantations, with an estimated annual loss exceeding 50 million metric tons. It is, therefore, projected that if the necessary actions are not taken, BSR disease can wipe out close to 860,610 ha of mature oil palm plantations in Malaysia by 2030 ( Olaniyi & Szulczyk, 2020 ). Several Ganoderma spp. viz. G. applanatum, G. boninense , G. zonatum , G. chalceum , G. lucidum , G. miniatocinctum, G. ryvardenii , G. pseudoferreu and G. tornatum have been linked to BSR disease on oil palm by early researchers ( Midot et al., 2019 ; Kumar et al., 2022 ; Kinge & Mih, 2011 ; Turner, 1981 ). Studies conducted by Chakrabarti et al. (2023) and Mohd Rakib et al. (2014) confirmed G. zonatum as the most prevalent fungal pathogen responsible for BSR and upper stem rot (USR) disease in oil palms, accounting for 71.7% of the total infected samples collected. Moncalvo (2000) also reported that G. boninense is the most virulent species, causing a high incidence of BSR disease in oil palm. Other researchers also confirmed this ( Bharudin et al., 2022 ; Khoo & Chong, 2023 ; and Susanto et al., 2005 ). All these reports indicate that BSR disease in oil palm is associated with varied pathogenic species of Ganoderma fungi. Therefore, to develop effective management strategies to protect Ghana's oil palms against BSR disease, further studies were needed to identify and confirm the actual Ganoderma species causing BSR disease on oil palms in Ghana. 2. Materials and methods 2.1 Study area Disease assessment was conducted in 30 oil palm growing communities in six districts in Central, Eastern and Western Regions of Ghana, two districts per region and five farming communities per district. For the purpose of this study, the selected districts surveyed were renamed as K29B, K37-1, K31-1, K37-2, K30, and K4. The K29B and K37-1 represent Denkyembour and Fateakwa districts from the Eastern region, K31-1 and K37-2 represent Nzema West and Mpohor districts from the Western region, while K30 and K4 represent Twifo-Atti Morkwa and Gomoa East districts from Central Region. 2.2 Climate of the study area The districts visited in Eastern region, lie within latitudes 6°.10871′ N to 6°.38431′ N and longitudes 0°.35047′ W - 0°.30′ W and 0°10′E to 0°30 E. They cover land areas ranging from 1500 km 2 to 2050 km 2 of the region. In the Western region, the two districts lie within latitudes 4.8520°N to 5.2 609°N and -2.2377° W to 1.5021°W and covered land areas ranging between 1,200 km 2 to 6,231 km 2 . Twifo-Atti Morkwa (K30) and Gomoa East (K4) districts from the Central region lie within latitudes 5°3.209′ N to 5°7.891′ N and longitudes 1°23.200′ W - 1° 54.360′ W. They cover land areas ranging from 1,160 km 2 and 5,231 km 2 of the region. All these regions are located within the semi-deciduous forest zone in Ghana. The areas are characterized by a double maxima rainfall pattern followed by a prolonged dry season. The minimum temperature during the study period (October 2022 to February 2024) ranged between 22.6°C and 25.2°C and the maximum varied between 31.4°C and 36.0°C. The relative humidity varied from 45% to 75%. 2.3 Field observation and assessment Intensive BSR disease scouting was carried out based on the single-point disease assessment using the standard operating procedure (SOP) of the Ganoderma census published by the Malaysian Palm Oil Board (MPOB, 2014). The process was done by counting the number of infected trees using a block of a hundred trees along a diagonal on each plantation for proper disease representation. A total of 3000 palms were assessed, 100 palms per plantation. 2.4 Percentage disease intensity Disease incidence was computed using modified version of Vicent’s formula ( Lekete-Lawson et al ., 2024 ). % BSR Disease Incidence ( DI ) on the field = Number of BSR infected oil palm per Plantation Total number of selected oil palm per Plantation × 100 Disease severity was scored on a scale of 0-4 ( Lekete-Lawson et al ., 2024 ) where, 0 = no symptom, and 4 ≥ 9 Maximum number of fruiting bodies per palm or foliar symptoms > 4 fronds/palm 2.5 Sampling and isolation Ninety symptomatic samples, including fruiting bodies (Basidiocarps) and Ganoderma sp. (Dikaryotic), were collected from infected life palms across all three regions ( Figure 1 ). Figure 1. Prevalence of BSR disease in selected Districts in Ghana. 2.6 Fungal culturing and isolation Media compositions were as follows: PDA for one litre of distilled water included: 39 g of Pre-mixed, Dehydrated PDA powder (Supplier: Thermo Fisher Scientific, Catalog code: CM0139B amended with 3 g of peptone (supplier: Clinichem Ltd, catalogue code: 70161), pH: 5. Malt extract agar (MEA: supplier: Thermo Fisher Scientific, Catalogue code: CM0059B); composition per liter of distilled water: 30 g of malt extract, 3 g of peptone, and 15 g of agar (Supplier: Millipore Sigma, Catalogue number: 05040); pH: 5.6. The diseased samples collected from both the infected oil palm trunk and the Basidiocarps were cut into approximately 1 cm pieces with a scalpel blade and rinsed three times in sterilized distilled water, surface sterilized in a 10% sodium hypochlorite (solution, and blotting on tissue paper ( Goh et al., 2014 ). The samples that were sterilized were aseptically inoculated onto PDA (powder (Supplier: Thermo Fisher Scientific, Catalog code: CM0139B) and incubated at 28±1 °C for four days and later sub-cultured on Malt Malt extract agar (MEA: supplier: Thermo Fisher Scientific, Catalogue code: CM0059B) till pure cultures of isolated fungi were obtained. The pure fungal cultures were maintained on Malt Extract agar (MEA) Malt extract agar (MEA: supplier: Thermo Fisher Scientific, Catalogue code: CM0059B) as stock cultures in Petri dishes in dark conditions at room temperature (28 ± 1°C) for further analysis. 2.7 Morphological identification Macro-morphological identification and confirmation were done using colony characteristics (shape, colour and texture of the mycelia) and micrograph description. Cultures from various plantations were grouped based on their morphological resemblances. There were three replications per isolate. Sporulation was assessed on glass slides by mounting a small portion of mycelia in sterilized distilled water with a blue stain and observed under a Leizer microscope of lens magnification of 40X. Mycelia, obtained from the various samples, were preserved in the biological incubation chamber (Thermostatic Cabinet ST 1 POL-EKO2; ST 1/1/1, Manufacturer: POL-EKO . Registered trademark ( ® , ™): POL-EKO ® sp.k of POL-EKO-APARATURA sp.j.) at 20 °C for 12 days, for pathogenicity tests and other clinical analyses. 2.8 Molecular identification and confirmation Infected BSR samples, fruiting bodies and isolated fungi were taken to CABI Microbial Identification Service, Bakeham Lane, UK for Molecular identification and confirmation. 2.8.1 Procedure as follows All original samples ( Ganoderma fruiting bodies and isolated fungi) were tested for purity. Molecular assays were conducted on all samples with nucleic acid as the template. A total of 29 basidiocarps and 24 Ganoderma pure cultures were investigated. Microzone, UK, proprietary formulation microLYSIS ® -PLUS (MLP) underwent rapid thermal cycling for cell lysis and release of DNA. Polymerase Chain Reaction (PCR) was used to amplify the rDNA in vitro after DNA extraction. The DNA quality of the PCR products was then evaluated. The samples were electrophoresis using a 2% agarose gel (Supplier of agarose gel: Thermo Fisher Scientific, Catalogue Number: UltraPure™ Agarose, 500 g – 165005001) and GelRed-stained (supplier: Biotium, Inc., Catalogue Number: GelRed ® Nucleic Acid Stain, 10,000X in water - 41003/41003-1). An additional PCR purification step was performed to eliminate excess dNTPs, primers, polymerase, and other constituents of the PCR mix, in order to obtain an extremely pure DNA template for sequencing. PCR amplicons were later characterized by sequencing with the BigDye ® Terminator v3.1 kit from Applied Biosystems (Life Technologies, UK,) and a pair of primers: ITS1 (TCC GTA GGT GAA CCT GCG G) and ITS4 (TCC GCT TAT TGA TAT GC GTAC) ( Gašparcová et al., 2017 ). The basidiomycete-selective primer GanET (GCGTTACAT GAG CGCAATACAA) was also used to prevent the potential co-amplification of contaminant non-basidiomycetous fungi. Sequencing reactions were carried out with the use of fluorescently tagged chain terminator dNTPs. Sample processing was executed with the AB 3130 Genetic Analyzer for the examination of the sequence of nucleotide bases (adenine, guanine, cytosine, and thymine) in the DNA oligonucleotide. Preliminary identification was made by matching the sequenced data with the sequences in the European Molecular Biology Laboratory (EMBL) database via the European Bioinformatics Institute (EBI). For verification, the samples were re-identified by ITS sequence analysis using the FASTA algorithm and the Fungus database from EBI and the BLAST algorithm with the NCBI standard database (Report on molecular work: https://figshare.com/s/86864d9d08393658fb20 ) (Accessed: 07.02.2025). 2.8.2.0 Phylogenetic analysis 2.8.2.1 Sequence alignment Sequences of newly identified species were analysed using standard BLAST searches in GenBank: [ Lekete-Lawson, (2025a) , ( https://doi.org/10.6084/m9.figshare.28389083 )] to identify more similar sequences. In addition to sequences obtained in this study, all sequences used for phylogenetic analysis were retrieved from GenBank: ( ftp://ftp.ncbi.nlm.nih.gov/pub/agarwala/indexed_megablast ). The sequence multiple alignments were done using the online version of MAFFT (v7.511) https://mafft.cbrc.jp/alignment/software/ and open access online version of Geneious Bioinformatics Software for sequence Data analysis R9 v. 2019.2.3 ( https://www.geneious.com/ ): Rozewicki et al. (2019) manually formatted with Clustal-Omega 1.2.4 (open access available: http://www.clustal.org/omega/ ) pairwise sequence alignment tools to minimise gaps and ensure proper alignment. 2.9 Pathogenicity test (Koch’s postulate) For the purpose of this study, three-weeks-old healthy germinated palm seeds of commercial standard crosses of (Dura × Pisifera, (D × P)) obtained from Ghana Sumatra Ltd were used for the pathogenicity test. A volume of 250 ml Beatson glass jars (R3/83 mm 4 Doz) containing a mixture of planting media (black soil, bio-cha, and treated plant biostimulant) at 2:2:1 100 g per glass was used. The jars were autoclaved for 45 minutes at 121°C. The mixtures were left to cool for three days. After three days, healthy germinated seeds with uniform growth were sown into the glass jar containing the sterilized growth medium for artificial inoculation ( Goh et al., 2014 ). A total of Hundred and forty-four (144) germinated palm seeds germinated palm seeds (three treatment and control, 6 seeds per replicate) were arranged in a randomised complete block design (RCBD) in the growth chamber and maintained for one week under laboratory conditions at CSIR-OPRI Plant Pathology laboratory, (6°.10871′N, 0°.35047′W and 0°10′E) with relative humidity and temperature regulated at 70 % and 28±1°C, respectively. 2.9.1 Inoculum preparation Research has shown that Ganoderma species are wood-decomposing fungi that thrive on decaying wood, which provides essential nutrients and moisture ( Kumar et al., 2022 ; Mangaiha et al., 2019 ; Chang et al., 2002 ). Thus, techniques by Angel et al. (2021) and Bivi et al. (2016) , with slight modifications, were used in Ganoderma inoculum preparation using Woodblocks made from Triplochitan scleroxylon (Wawa), which was previously tested in preliminary studies (Anonymous, 2024-unpublished) to be an effective growth medium for Ganoderma fungi. The Wawa Woodblock (WWBs) incubation time was 15 days ( Breton et al., 2005 , 2006 ). 2.9.2 Artificial inoculation of germinated oil palm seeds Inoculation of germinated seeds was performed in 250 ml Beatson glass jars (R3/83 mm 4 Doz) with WWBs previously colonized by Ganoderma isolates. Glasses (components) were incubated according to the method described by Lo et al. (2023) with slight modification. The set-up was maintained under lab conditions with relative humidity and temperature regulated at 70% and 28±1°C, respectively. The infection process was monitored and recorded daily. Germinated seed nuts with WWBs without inoculum served as a negative control. Re-isolation of inoculated fungi from the destructive samples was performed following the techniques by Agrios (2005) and Idris et al. (2006) . The roots were treated and plated on Malt extract agar and subcultured until a pure culture was obtained and identified by Lo et al. (2023) 2.9.3 Symptoms recording Disease development based on external and internal symptoms was recorded every three days. External symptoms were based on visual estimation of the proportion of tissues damaged by inoculated fungi, using the scale established by Breton et al. (2006) . Internal symptoms were recorded by splitting the inoculated seed nuts in two cross-sections and through the root. Other symptoms which were not visible were confirmed using molecular tools. All the signs and symptoms recorded were used for disease rating. 2.9.4 Disease rating Disease rating in vitro was computed as: % BSR Disease Incidence ( DI ) per germinated seed nut = Number of infected oil palm seed nuts Total number of inoculated seed nuts × 100 2.10 Data analysis The results for interpreting pathogenicity tests were based on the percentage of infection on test seed nuts. These were subjected to an ANOVA, and treatment means were separated with the least significant difference (LSD) at 5% after rating (R-Software data analysis: Lekete-Lawson, E. (2025a) https://figshare.com/authors/Emmanuellah_Lekete-Lawson/20618918 ). The data on different parameters were also compiled and analysed using the R-Stat version 2021 statistical package ( R Core team (2021) https://www.R-project.org , accessed 10/8/2024). The LSD at 1% was also used to separate the means of treatment fungi and the control. 3. Results 3.1 Field observation and percentage infection Out of 3000 palm trees assessed, 341 palms were identified as Ganoderma- infected palms representing 11.3% of disease incidence. However, the level of infection differs. The number of fruiting bodies (basidiocarps) counted on each infected palm varied between 2 and 274, and severity scores were between 2 and 4, with 4 being the highest recorded on the 9-year-old palm. District K29-B in the Eastern region showed the highest percentage of infection, with 161 infected palms and 274 basidiocarps. District K4 has the least, with 2 infected palms and 4 basidiocarps representing a severity score of one. This study revealed a higher infection rate in the Eastern region compared to the other two regions surveyed. Figure 1 shows the total number of palms affected and the severity score range of BSR disease in the selected oil palm growing Districts in Ghana. 3.2 Physiological characteristics of Ganoderma symptoms Basal stem rot disease observed on the fields depicts the basidiocarps which first appeared as small white buttons ( Figure 2B ) of tissue on the stem and later developed into a bracket shape mainly on the base ( Figure 2Dii ) or at the upper side ( Figure 2Di ) causing Basal Stem Rot (BSR) or Upper Stem Rot (USR) disease. These basidiocarps are woody, perennial and dimidiate; their cap (pileus) is concave, and some are circular with rough and irregular margins/blades (lamellae/gills). Pileus measured from 4-17 cm in diameter ( Figure 2Di ); and 7 cm to 19 cm in length. Surface glabrous; crisped; shiny; dry and smooth. Their upper surfaces also took on a variety of white to yellowish-brown colours and was with concentric zonations ( Figure 2Dii ). Also, the underside/gills are white ( Figure 2 ), gills adnexed, crowded (8-14 attached lamellae) with 8 series of lamellae, narrow and crisped. Most of the BSR basidiocarps lacked a stipe. However, in some upper-stem rot mushrooms, a short, wide, and noticeable stipe was present ( Figure 2Di ). This stipe was lateral, compressed, and uniform in width, with a bulbous base and distinct longitudinal striations. The hymenophore is woody and dark-brown with no volva. No basidiospores, basidia, and basidioles were observed. The substrates were observed in live mature oil palms ( Figure 2 ). Figure 2. Ganoderma-infected palms: (A) symptomatic samples for isolation, (B) Initial stage of infection and pinhead of the Ganoderma mushroom, (C) Fractures and collapsed Ganoderma infected palm, (Di) Upper Stem Rot infection (Dii) Basal Stem Rot infection. Many of the heavily infected palms were seen full of basidiocarps growing from the base ( Figure 2 A, B, C & Dii ; some deteriorated and others fractured and collapsed ( Figure 2C ). 3.3 Macromorphological Characteristics of culture of Ganoderma fungi 3.3.1 Isolation and identification of causal agent(s) of the BSR disease on oil palm Out of 90 symptomatic samples collected from the field, 60 of them were screened for fungal isolation. 29 categories of fungal cultures were obtained and identified, 24 of them were identified as suspected Ganoderma fungi, and grouped into three (GA, GD2B and TD2B) ( Figure 4 ). Other fungi isolated and identified include Trichoderma, Xylaria, Fusarium , Phytophthora , and others (moulds; Rhizopus , Aspergillus spp.). These fungal isolates were grouped based on their macromorphological characteristics and micrograph features. They were further ranked based on their taxonomy and percentage of occurrences ( Figure 3 ). Figure 3. Common fungal species associated with Ganoderma-infected palms. Overall, suspected Ganoderma fungi were identified 24 times, representing 66% of the total isolates. This was followed by Trichoderma sp. (13%) with saprophytic fungi (others) recording the least (2%). These suspected Ganoderma fungi were also categorised into three based on colony similarities in culture media, growth rate and their place of origin. Isolates GA from the Eastern, GD 2 B from the Central and TD 2 B from the Western regions ( Figure 4 ). Figure 4. A) Pure culture of suspected Ganoderma sp. B) Micrograph of the isolates (image magnified 40X). All cultures in Figure 4A were produced simultaneously. GA had the fastest growth rate, followed by TD 2 B and GD 2 B. These three groups of the Ganoderma isolates produced white, velvety, fluffy and cottony colonies on Malt extract agar medium ( Figure 4A ). GA colonies appeared white, stranded, cottony and fluffy ( Table 1 ). GD 2 B produced white and stranded mycelium, velvety with concentric cottony and hyphal-knot structure ( Table 1 ). TD 2 B appeared white, fluffy, cottony and velvety but turned yellowish-brown as it progressed ( Figure 4 ). Their mycelium micrograph also shows hollow, septate and clamp connections ( Figure 4B ) when viewed under a compound microscope with a magnification of 40X. No spore was seen on the artificial medium. Table 1 shows macromorphological features of the three groups of Ganoderma fungi isolated. Table 1. Macromorphological features of pure cultures of suspected Ganoderma isolates. Isolate group Classification Colony characteristics GA Ganoderma sp. The colonies are whitish and radial, mycelium covers the entire Petri dish, creating dense and fluffy colonies in the centre. Colonies are continuous, radial, and have a cotton-like texture with snow-white (niveus) and later whitish (albidus) colours. They grow radially and form thinner and denser zones, with the centre becoming yellowish. GD2B Ganoderma sp. The colonies are whitish, have a cotton-like texture and a strong leather-like appearance, hard to cut with a scalpel. They have slightly fuzzy margins and a snow-white (niveus) colour similar to GA isolate. After 12 days of inoculation, the surface becomes floccose with flaky mycelial formations, turning snow powder-like snow-white (niveus) and whitish (albidus) colours. stranded mycelium, velvety with concentric cottony and hyphal-knot structure. TD2B Ganoderma sp. Colonies are whitish, dense, and thick after 8 days of inoculation. They take a yellowish shade, form radial depressions, and become dense and fluffy. Colony turns yellowish/brownish at the margin after covering the entire Petri dish. 3.4 Confirmation of pathogenicity test of isolated fungi 3.4.1 Source of inoculum The study showed that the three isolated Ganoderma fungi (GA, TD 2 B and GD 2 B) were able to colonise the inoculated WWBs. The first sign of growth was observed after seven days of inoculation with GA isolate, TD 2 B isolates after 12 days and GD 2 B appeared after 16 days of inoculation. The total incubation time of the WWB inoculum was 17 days. The growth chamber was maintained at a temperature of 28±1°C and relative humidity of 60-75% suitable for the fungal growth. 3.4.2 In vitro artificial inoculation This research presents an in vitro artificial inoculation test on oil palm germinated seed nuts, in a controlled environment resulting in the first symptoms development within 2-3 weeks. The first symptoms on inoculated seeds appeared after eight days of inoculation and differences between the inoculates and inoculated germinated seeds and control ( Figure 5dii ) became noticeable around 14 to 16 days ( Figure 5aii and Figure 5bii ). Figure 5. (i) Source of inoculum-Wawa woodblock (WWBs) (ii) Confirmation of pathogenicity of Ganoderma isolates (d ii) control seed nuts. After 21 days of artificial inoculation, germinated seed nuts inoculated with WWBs inoculum showed physiological symptoms of disease including root decay ( Figure 5(aii and cii) , necrosis ( Figure 5(aii , bii and cii )), leaf withering ( Figure 5(aii , and cii )), decaying bole tissue ( Figure 5bii ) and other lesions that were absent in the control group of seed nuts ( Figure 5dii ). 3.4.3 External and internal symptoms evaluation The percentage infection was determined by considering both external and internal disease indices ( Table 2 ). The external infections were recorded based on visual symptoms observed in the inoculated seedlings while internal infections were evaluated based on endophytic symptoms analysis and re-confirmation of pathogenicity using molecular tools. Table 2. Statistical analysis of pathogenicity of Ganoderma isolates on germinated oil palm seed nuts. Ganoderma isolates Estimate Std. Error t-value Pr(>|t|) p-value T1: GA 72.20000 8.221680 7.6219 0.0000004*** T2: GD 2 B 44.43333 5.059787 5.9914 0.0003237* T3: TD 2 B 61.10000 6.957682 9.3843 0.0000053** Control (no inoculum) 13.90000 2.238480 3.6722 2.51120 3.5 Analysis of pathogenicity Table 2 shows the results of the pathogenicity tests of different Ganoderma isolates (T1:GA, T2:GD 2 B, T3:TD 2 B) from BSR-infected oil palms in Ghana. Each isolate (GA, GD 2 B TD 2 B,) and the control group were analyzed with estimated effects, standard errors, t-values, and p-values to measure their relative impact on the test seed nuts. Out of 144 seedlings tested, 58.3 % were infected. Data showed that GA fungi from the Eastern Region exhibited severe pathogenic effects (t-value: 8.221680) on the germinated seed nuts and caused 26.1% of seedling deaths ( Figure 6 ). Its treatment against the control showed a highly significant (p<0.01) difference with adjusted p-value (p adj) of (0.0000004)***). Figure 6. Record of percentage seedlings death after artificial inoculation. 3.5.1 Pathogenicity variation and treatment comparisons The treatment GD 2 B isolates showed a significant increase of 44.433 units with a t-value; (5.059787) and a p-value of 0.0003237 compared to the control. Isolates from the Western region (TD 2 B) showed a relatively strong effect (t-value = 6.957682), on the treated seed nuts compared to the control group. GD 2 B shows a decrease of -27.77 difference with a weak pathogenicity effect (p-value: 0.0233249), confidence Interval (lower, upper): (-52.35, -3.19) and a low percentage (2 %) seedling death ( Figure 6 ) ( Table 3 ). Isolate TD 2 B shows a decrease in virulence effect (-11.1 units) compared to the GA isolates, with an adjusted p-value (p adj): of 0.5952326 and a confidence interval (lower and upper): (-35.68, 13.48) of which is not significant. Comparing the pathogenicity effect of TD 2 B and GD 2 B isolates, TD 2 B shows an increase of 16.67 difference with the confidence Interval (lower and upper): (-7.91, 41.25) compared to GD 2 B, but this difference is not significant (p-value = 0.2604245). Table 3. Pathogenicity variation among the Ganoderma fungi tested. Treatment diff lwr upr p adj T1(GA)-Control 72.2 47.6207 96.7793 0.00004 T2(GD 2 B)-Control 44.4333 19.854 69.0126 0.00032 T3(TD 2 B) -Control 61.1 36.5207 85.6793 5.3E-06 T2(GD 2 B)-T1(GA) -27.7767 -52.346 -3.18736 0.02332 T3(TD 2 B) -T1(GA) -11.1 -35.6793 13.4793 0.59523 T3(TD 2 B)-T2(GD 2 B) 16.6667 -7.91264 41.246 0.26042 3.5.2 Symptoms recording The highest external symptoms (62.21%) and seedling death were observed in the seed nuts inoculated with GA and TD 2 B fungi ( Figure 6 ) and the lowest severity (4.23 %) was observed in the GD 2 B treated seed nuts. None of the other isolated fungi ( Trichoderma , Xylaria , Fusarium and Phytophthora and saprophytes (moulds) produced similar BSR symptoms when tested under Koch’s postulate. 3.6 Molecular characterisation and confirmation 3.6.1 Real-time PCR assay A total of 53 samples (24 pure culture-based isolates and 29 Ganoderma Basidiocarps) were amplified. ITS/18rRNA gene sequencing and MLST (multilocus sequence typing) were subsequently used to further characterize and identify the Ganoderma fungi isolated from the initial infected samples. DNA extracted from the basidiocarps and the Ganoderma isolates from the infected oil palm were amplified using ITS/GanET gene sequencing primer pair. Multilocus sequence typing (MLST) produced a 320 bp fragment and sequenced as the DNA amplicon. A single separation line was observed in Temperature gradient gel Electrophoresis (TGGE) ( Figure 7 ) at the standard band corresponding to the ITS regions of Ganoderma sp. showing consistent results. There was no significant difference between the bands produced by the 29 genotypes of basidiocarps ( Figure 7B ) and those of the culture-based samples ( Figure 7A ) except for sample numbers (20) and (35) whose bands were very faint. Figure 7. (A) PCR amplification of selected Ganoderma basidiocarps and (B) PCR amplify from culture-based isolates. 3.6.2 Sequence results The ITS/ITS4/GanET gene sequence primer pair from the amplified samples showed top matches of >97% to sequences assigned to members of the genus Ganoderma. Fully (and validly) named matches of >98% included 99.3% identity to three sequences of Ganoderma sp. (HM138671; HM138670 and HM138672) generated from strains assigned to Ganoderma ryvardenii (syn. Ganoderma ryvardense ). Sequence HM138671 is from the type strain of G. ryvardenii. (HKAS58053). A match of 99% was also obtained to sequence MN809325 from Ganoderma wiiroense strain LDCMY02. Other best matches notified to published strains were only at 92.0-95.6% identity to sequences assigned to G . boninense. Out of total sample amplified, 89% of the sequence obtained matches G. ryvardenii, 7 % to G. wiiroense and 4 % matches G. boninense. 3.6.3 Proof of Koch’s postulate 3.6.3.1 Re-isolation and Molecular confirmation of Ganoderma fungi after pathogenicity test Amplification of the DNA extracted from the bole and root tissues of the artificially inoculated oil palm seedlings using the ITS1/ITS4 primer pair produced similar base pairs (320 bp) of DNA fragment, which was the same size sequenced as the amplicon of DNA ( Figure 7B ) from Ganoderma pure culture ( Figure 4 ). The identity of the fungi was confirmed again as Ganoderma ryvardenii via the sequencing of the ITS primer pair amplified products. All the isolates obtained after artificial inoculation had their ITS region sequences that were split into a single line and belonged to the taxon Ganoderma which has 99.3% of its gene sequence similarities with the strain of G. ryvardenii. 3.6.4 Phylogenetic results The standard band quality of the overall amplification most likely corresponds to a novel species of Ganoderma. The phylogenetic analyses using ITS sequences and their accession numbers confirmed the identification of a new Ganoderma species ( Figure 8B ). Additionally, the majority of the obtained sequences were associated with Ganoderma ryvadenii with their corresponding accession numbers as indicated by the arrows in Figure 8B . Figure 8. (A) Phylogenetic relationship between different species of Ganoderma fungi associated with BSR on oil palm (B) the accession numbers of the novel species (arrowed) of Ganoderma fungi in this study. 4. Discussion 4.1 Field observation and percentage infection Field observations and percentage infection rates of Ganoderma in the selected oil palm plantations provide important information about the disease's spread, effects, and mechanism of transmission. Although BSR was initially found in palms older than 25–30 years old, it has recently been reported that the disease can affect younger palms, including those as young as one year old ( Zakaria, 2023 ). This study showed a higher infection rate in the Eastern region than in the other two Regions surveyed. Despite the low disease incidence in the selected districts compared with the total number of palm trees analysed, the rate of disease spread was of grave concern. A similar observation was made by Lekete-Lawson et al. (2024) in a separate study on oil palm in Ghana. 4.2 Physiological characteristics of Ganoderma symptoms The physiological features portrayed by the pathogen in this study, shared similar characteristics of Ganoderma species described by Bharudin et al. (2022) and Chong et al. (2017) . According to these authors, basal stem rot (BSR) in oil palm is a disease caused by bracket fungi, producing basidiocarps which are perennial hard-knot and pinhead-like when immature; fan-shaped and large when mature; with irregular margins/blades (lamellae/gills). They are called Ganoderma fungi and are members of the phylum Basidiomycota and the family Ganodermataceae. Hushiarian et al. (2013) and Pilotti (2005) also reiterated that the basidiocarp is a typical characteristic feature of Ganoderma fungi. These basidiocarps are lignicolous and leathery and sometimes grow in a hoof-like form on the trunks of infected palms, just as observed in the present study ( Figure 2 ). Some of the heavily infected palms that were seen full of basidiocarps were rotten, fractured and collapsed as a result of the infection. Similarly, Siddiqui et al. (2021) and Josephin et al. (2024) observed that after the oil palm tree becomes infected, the fungus quickly spreads throughout the trunk tissues, causing the internal structures to deteriorate leading to the collapse and the death of the infected palms. 4.3 Isolation and identification of causal agent(s) of the BSR disease on oil palm 4.3.1 Macromorphological characteristic of culture of Ganoderma fungi The isolated fungi were grouped based on their macromorphological characteristics and micrograph features. They were further ranked based on their taxonomy and percentage of occurrences. Research has shown that most Ganoderma species identified in the past were based on morphological characteristics ( Latiffah & Ho, 2005 ). However, studies by Mohd Rakib et al. (2014) showed that Ganoderma species are genetically heterogeneous and different based on their geographical origin, thus resulting in genetic variations even within the same species ( Hong et al., 2001 ). This was evident in the present study when the macromorphological characteristics of colonies of Ganoderma fungi isolated from the three regions appeared different. The study showed slight differences in culture characteristics among the three Ganoderma isolates (GA, GD 2 B and TD 2 B) regarding their colony colour, texture and growth rate. The white and stranded mycelium, velvety with concentric cottony and hyphal-knot structure produced by GD 2 B isolates were similar to what was found in the studies conducted by Beck et al. (2018) and Badalyan et al. (2015) on several Ganoderma species, in which some species of Ganoderma appeared as cottony texture with powdery radiating parallel hyphae which were white and later changed to lemon yellowish. TD 2 B appeared white, fluffy, cottony and velvety but turned yellowish-brown as it progressed. Mohanty et al. (2011) discovered that some Ganoderma species age as their colonies turn yellowish to brown. The varied growth rate observed among the isolates with GA exhibiting the fastest growth rate could depend on several variables, including the culture's duration, the media's composition, the ambient temperature and the media composition. According to Gáperová and Petrýdesová (2009) several elements, including light, temperature, medium volume, nutritional component ratio, and others, might influence the properties of mycelia under study. Although our findings suggest that the macromorphological traits of cultures and the mycelium's growth rate may be utilised to identify specific Ganoderma species, more research is needed to distinguish individual species. 4.4 Confirmation of pathogenicity test of isolated fungi 4.4.1 Source of inoculum Triplochiton scleroxylon (Wawa woodblocks-(WWBs)) used for inoculum in this study helped to retain the survival of isolated fungi for the inoculation. The studied Ganoderma fungi (GA, TD 2 B and GD 2 B) were able to colonise the inoculated WWBs but at varied degrees of myceliation. According to Mangaiha et al. (2019) , woody debris (inoculum) plays an important role in the long-term survival of Ganoderma fungi. They enhance the resistance of the growth of Ganoderma fungi against environmental stress ( Loyd et al., 2018 ). Woodblocks (WWBs), as a source of inoculum, produced successful infection on oil palm germinated seeds. Although, research revealed that Rubberwood block (RWB) is an ideal substrate that improves the long-term survival, biological yield and productivity of Ganoderma fungi ( Breton et al. (2006) ; Alexander, (2017) and Nusaibah et al. (2016) ), however, an experiment conducted by Angel et al., (2021) showed that if RWB becomes scarce, other substrates, including sawdust and wood chippings, could be used as a source of inoculum, through non-invasive tissue culture techniques. This was confirmed in this study when Wawa woodblock (WWB) as a substrate and inoculum source, was able to establish Ganoderma infection on the oil palm germinated seed nuts marking the first successful attempt at using the tissue culture technique for establishing Ganoderma infections on oil palm germinated seed nuts in Ghana. Nonetheless, other isolated fungi such as Trichoderma , Phytophthora, Fusarium , Xylaria, and others (moulds) were not able to colonise the substrate (WWBs). 4.5 In vitro artificial inoculation Breton et al . (2005 , 2006 ) showed a positive correlation between germinated seeds and 3-month-old seedlings in the discrimination between progenies for their susceptibility to Ganoderma. Although the formation of Ganoderma basidiocarps was not observed in this experiment probably due to the short time allocation and the substrate used, the disease symptoms, displayed by GA, GD2B and TD2B isolates were similar to those of a typical BSR disease in young seedlings. Similar features were also observed by Angel et al. (2021) after post-inoculation of plantlets with sawdust inoculum which displayed physiological symptoms of the disease that resembled those of a typical BSR infection. Unlike inoculation tests on 3-month-old seedlings that had been conducted by previous researchers including Idris et al. (2004) and Rees et al . (2007) , the results of this study have shown that in vitro inoculation test can be used as an alternative to reduce the age needed for inoculation techniques, shortening inoculation duration, and ensure consistency in experimental results. 4.6 Analysis of Pathogenicity 4.6.1 Pathogenicity variation The results suggest robust evidence against the null hypothesis, indicating that all isolates tested in this study exhibit a higher degree of pathogenicity. However, some disparities were observed in the aggressiveness of isolated Ganoderma fungi from the Eastern, Western, and Central regions of Ghana comparing their pathogenicity against control. GA isolate has the most virulent pathogenic effect causing the highest disease infection and death of the test seed nuts. This points to the fact that GA is likely responsible for the majority of BSR infections in the selected oil palm plantations in Ghana. The variations in the level of pathogenicity identified among the Ganoderma isolates in this study correspond to findings by Lo et al. (2023) , Goh et al. (2014) , Mohd Rakib et al. (2014) and Breton et al. (2006) . These individual authors also observed some differences in the degree of pathogenicity of Ganoderma pathogens on oil palms from different geographical locations. For instance, Lo et al. (2023) and Mohd Rakib et al. (2014), observed that there were some variations in aggressiveness within the species of Ganoderma boninense obtained from infected oil palms at different estates in Sarawak. Breton et al. (2006) discovered that G. boninense isolates from different plantations in Indonesia had varying levels of aggressiveness. Likewise, in Malaysia, the transmission rate of Ganoderma disease varies by estate, potentially indicating differences in G. boninense aggressiveness across isolates from different regions ( Goh et al., 2014 ). Mohd Rakib et al. (2014) also revealed similar observations about the significant variation in the aggressiveness levels of Ganoderma species isolated from different areas. The authors observed that G. boninense and G. zonatum isolated from Betong and Miri differed in their aggressiveness. This was demonstrated when all the Ganoderma isolates obtained from various oil palm growing areas in Ghana showed varied responsiveness in their disease establishment. The gradual decline of growth among the germinated seed nuts after being inoculated with Ganoderma isolates was observed. From day 14, the virulence factor of GA isolates from the Eastern region could be clearly distinguished from the others (TD 2 B and GD 2 B). The effect of the isolated fungi appeared after 15 days of incubation with initial infection of lesions forming on the roots of the inoculated seed nuts. This corresponds with the characteristic infection pattern by Ganoderma spp. as reported by Rees et al. (2007) and Breton et al. (2006) . According to Rees et al. (2007) , Ganoderma infection usually starts from the root system of the infected palms when in direct contact with the inoculum. Isolate GA has the strongest pathogenic effect on the test seed nuts, compared to isolate TD 2 B. However, the pathogenic effect of TD 2 B isolate is still highly significant (p<0.05). The highest external symptoms and seedling death was observed in the seed nuts inoculated with GA and TD 2 B fungi whereas the lowest severity was observed in the GD 2 B treated seed nuts. Apparently, none of the other isolated fungi ( Trichoderma , Xylaria , Fusarium and Phytophthora and saprophytes (moulds) were able to produce similar BSR symptoms observed on the original samples when tested under Koch’s postulate. 4.7 Molecular characterisation and confirmation 4.7.1 Real-time PCR assay Despite the weak fragment shown in the band formation of samples 20 and 35, probably due to PCR inhibitors, the standard band quality of the overall amplification most likely corresponds to a novel species of Ganoderma. He et al ., (2022) predicted that approximately 1.4 to 4.2 million species of Basidiomycota will be discovered worldwide by 2030, as over 54,000 have already been reported. Since the coming of the molecular age, several species previously described morphologically are now redefined as new taxa in addition to the finding of new taxa. To this effect, there are now over 36,000 Basidiomycota species, according to a comprehensive systematic survey using molecular data ( Begerow et al. 2018 ). The molecular tools employed in this experiment allowed us to differentiate between numerous fungal species of Ganoderma disease that are present in Ghana's oil palm fields. Sequences of other phylogenetic markers useful in the phylogeny of Ganoderma fungi supported our hypothesis that the isolates most likely represent a novel species of Ganoderma. For instance, the majority of the ITS sequences of Ganoderma sp. [HM138671; HM138670 and HM138672] generated from strains assigned to Ganoderma ryvardenii (syn. Ganoderma ryvardense) HM138671; HM138670 and HM138672 published in Mycosphere 2 (2): 179–188. Of these, sequence HM138671 is from the type of strain of G. ryvardenii. (HKAS58053). There was a single match of 100% to sequence MN809325 from Ganoderma wiiroense strain LDCMY02 but this has not been published in a peer-reviewed publication and is quite distinct from other sequences of G. wiiroense in GenBank, so it is discounted here. Thereafter, the third-best matches to published strains were also identified to sequences assigned to G. boninense. Additionally, the sequence alignment in phylogenetic tree building also revealed high similarities among Ganoderma strains identified in Cameroon (HKAS58053) and the new strains identified in Ghana (HM138671; HM138670 and HM138672). As such the phylogenetic analyses based on ITS sequences and their accession numbers confirmed the novelty of this Ganoderma species on oil palm in Ghana. This finding enhances our understanding of the diversity and distribution of specific Ganoderma species within African mycobiota. This also confirms the initial discovery made by Kinge and Mih (2011) , who first reported the pathogen on oil palm in Cameroon and gave the species name as Ganoderma ryvardense with the specific epithet in honour of Lief Ryvarden, a distinguished mycologist who has contributed significantly to genus Ganoderma and African mycobiota. (Mycosphere 2 (2): 179–188). Evidently, this is the first report of G. ryvardenii causing BSR disease on oil palm in Ghana and possibly second in West Africa. 5. Conclusion The study uses molecular techniques to identify and confirm the Ganoderma species responsible for causing BSR disease in selected oil palm plantations in Ghana. The sequences of other phylogenetic markers useful in identifying and confirming Ganoderma fungi supported our hypothesis that the isolates most likely represent a novel species of Ganoderma. Thus, this study underscores the importance of using molecular techniques for accurate identification. It highlights the genetic diversity and morphological variability of Ganoderma species on oil palm in Ghana forming a crucial understanding of their role in causing BSR disease on oil palm. The fungus was first identified as Ganoderma sp. based on the physiological features (Fruiting bodies/basidiocarps), disease symptoms, culture morphological characteristics and later confirmed through Koch’s postulate, molecular amplification of internal transcribed spacer (ITS sequence) and basidiomycete-selective primer (GanET) to obviate the potential co-amplification of contaminant non-basidiomycetous fungi. As such, these findings provide baseline information for future studies regarding the emergence of new species of Ganoderma on oil palm in Ghana. Data availability statement Figshare: Report on Basal Stem Rot (BSR) disease on oil palm in Ghana, DOI: https://doi.org/10.6084/m9.figshare.28329830.v1 ( Lekete-Lawson, 2025b ) The project contains following underlying data: • Extracted files • Gano data analysis. 1R • Book 1 Gano. Raw. Data are available under the terms of the Creative Commons Attribution 4.0 International license (CC-BY 4.0). The data generated in this study have been deposited in the Figshare repository and can be accessed at [DOI: 10.6084/m9.figshare.28329830 ]. Some of the article’s data (Accession numbers) can also be found in the following Nucleotide database [NCBI Blast: gb/HM138670/ https://doi.org/10.6084/m9.figshare.28389083 ], others were retrieved from the following public domain resources: The dataset includes raw sequencing data, processed data and Metadata (Map of districts surveyed, Treatment data on pathogenicity, R-Software data analysis and Ganoderma molecular work). The data are licensed under a CC-BY 4.0 license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Further details are available from the corresponding author upon request through the link: https://figshare.com/authors/Emmanuellah_Lekete-Lawson/20618918 . https://doi.org/10.6084/m9.figshare.28329830.v1 Extended data Figshare: Additional dataset to help with the journal contribution, DOI: https://doi.org/10.6084/m9.figshare.28389083.v1 ( Lekete-Lawson, 2025a ) The project contains the following extended data: • jpg Districts surveyed in the course of study • sequences used 1 • Accession numbers retrieved • sequence blast doc. TGCGGAAGGATCATTATC • NCBI Blast_gb_HM138670_ Data are available under the terms of the Creative Commons Attribution 4.0 International license (CC-BY 4.0). References Agrios GN: Plant Pathology. 5th ed.Burlington, USA: Elsevier Academic Press; 2005; p. 922. 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Reader Comment 04 May 2025 Robert Paterson , Centre for Biological Engineering, University of Minho, Braga, Portugal 04 May 2025 Reader Comment Comment on Lekete-Lawson et al. (2025) By: Robert Russell Monteith Paterson University of Minho, Portugal [email protected] I was interested to read the pre-review paper on Ganoderma from oil ... Continue reading Comment on Lekete-Lawson et al. (2025) By: Robert Russell Monteith Paterson University of Minho, Portugal [email protected] I was interested to read the pre-review paper on Ganoderma from oil palm in Ghana (Lekete-Lawson et al. 2025) having published quiet extensively in the field in other countries. It is worth reviewing the current situation. G. boninense is considered the basal stem rot (BSR) fungus in Southeast (SE) Asia and Oceania where Indonesia and Malaysia are by far the largest producers of palm oil. G. zonatum is also considered to cause BSR and a paper recently claimed that G. zonatum is the cause of BSR in Colombia, South America. G.ryvardenii is related closely to, but distinct from, G boninense and G. zonatum . Clearly, G. zonatum and G. boninense are also closely related. My first concern when reading Lekete-Lawson et al. (2025) was the PCR analysis and subsequently I detected other problems. PCR In relation to the PCR, in the text it says Figure 7A is for the culture based samples but these are Figure 7B in the legend to the image of the gel. Figure 7 B is the text is for basidiocarps but these are 7A in the legend to the figure. Which is it? Also, there is no information about the strain numbers that represented tracks 1 to 24 in the figure of the gels. These should be provided. It is all very confused. Also, there appeared to be a faint band in the blanks for samples 1 to 24 which were at the same intensity for track 20. Presumably the blanks were contaminated. Were tracks 20 and 35 from the same specimen and strain? Was there a procedure for identifying inhibition of the PCR? I know that an internal amplification control (IAC) to assess inhibition is recommended for at least some PCRs. IAC would provide information on inhibition or could it be that the samples represented by 20 and 35 were not what you thought they were? Why did you not run known relevant Ganoderma species for comparison with the other specimens? This would provide information on the novelty of your specimens. Overall, there are considerable problems with the presentation of the PCR results. Other concerns are as follows: General A strain and specimen list is necessary with accession numbers. Are you saying G. ryvardenii is the only species involved in BSR in Ghana? What is the new species you say you have discovered? This is unclear. A considerable amount of this paper is taken from a previous paper (Lekete-Lawson et al. 2024). This paper should be discussed in the introduction and cited properly. And any duplication needs to be removed. Somebody who is excellent at writing scientific papers in English should be asked to rewrite the paper. Abstract Palm oil is not necessarily the foremost vegetable oil. For example, soy oil is arguably more important. The abstract is vague. What do you mean by using nucleic acid as a template? The English is poor, e.g. “BSR is characterised by stem decay large-perennial, woody brackets basidiocarps”. What does this mean? The conclusion does not make sense to me at all. Introduction India is only a small palm oil producing country in contradiction to what you have written. You say Africa produces 3% oil palm and then West Africa produces 4%. Which is it? G. boninense is the most serious disease of oil palm in SE Asia and Oceania. In Africa it is considered that fusarium wilt is the most serious fungal disease. G. boninense is accepted as the fungus responsible for BSR in SE Asia at least. You need to mention that G. ryvardenii was identified in Cameroon before and give the citation. Material and Methods Figure 1 should be in the results. In the sampling and isolation section, what does “including fruiting bodies (Basidiocarps) and Ganoderma sp. (Dikaryotic)” mean? How were specimens tested for purity? What does “Microzone, UK, proprietary formulation microLYSIS®-PLUS (MLP) underwent rapid thermal cycling for cell lysis and release of DNA” mean? GanET is G. boninense specific (or better, a palm clade specific) primer. It is not basidiomycete specific. Please correct this. Results Did you test the difference between upper stem rot specimens and basal stem rot specimens? These have been considered as different species before. Phytophthora is not considered to be a fungus. It is an oomycete. The Phylogenetic results section is poor with inadequate information. Discussion Cameroon is not in west Africa. References Lekete-Lawson E, van der Puije GC, Osekre EA and Ackah FK. First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved with reservations]. F1000Research 2025, 14 :413 ( https://doi.org/10.12688/f1000research.161972.1 ) Lekete-Lawson, E, Osekre, E.A. van der Puije, G., Swanzy, et al. 2024. Prevalence of basal stem rot disease caused by Ganoderma sp. on oil palm ( Elaeis guineensis Jacq) in Ghana. Journal of Science and Technology, Special Issue No 1, pp 47 – 61. Comment on Lekete-Lawson et al. (2025) By: Robert Russell Monteith Paterson University of Minho, Portugal [email protected] I was interested to read the pre-review paper on Ganoderma from oil palm in Ghana (Lekete-Lawson et al. 2025) having published quiet extensively in the field in other countries. It is worth reviewing the current situation. G. boninense is considered the basal stem rot (BSR) fungus in Southeast (SE) Asia and Oceania where Indonesia and Malaysia are by far the largest producers of palm oil. G. zonatum is also considered to cause BSR and a paper recently claimed that G. zonatum is the cause of BSR in Colombia, South America. G.ryvardenii is related closely to, but distinct from, G boninense and G. zonatum . Clearly, G. zonatum and G. boninense are also closely related. My first concern when reading Lekete-Lawson et al. (2025) was the PCR analysis and subsequently I detected other problems. PCR In relation to the PCR, in the text it says Figure 7A is for the culture based samples but these are Figure 7B in the legend to the image of the gel. Figure 7 B is the text is for basidiocarps but these are 7A in the legend to the figure. Which is it? Also, there is no information about the strain numbers that represented tracks 1 to 24 in the figure of the gels. These should be provided. It is all very confused. Also, there appeared to be a faint band in the blanks for samples 1 to 24 which were at the same intensity for track 20. Presumably the blanks were contaminated. Were tracks 20 and 35 from the same specimen and strain? Was there a procedure for identifying inhibition of the PCR? I know that an internal amplification control (IAC) to assess inhibition is recommended for at least some PCRs. IAC would provide information on inhibition or could it be that the samples represented by 20 and 35 were not what you thought they were? Why did you not run known relevant Ganoderma species for comparison with the other specimens? This would provide information on the novelty of your specimens. Overall, there are considerable problems with the presentation of the PCR results. Other concerns are as follows: General A strain and specimen list is necessary with accession numbers. Are you saying G. ryvardenii is the only species involved in BSR in Ghana? What is the new species you say you have discovered? This is unclear. A considerable amount of this paper is taken from a previous paper (Lekete-Lawson et al. 2024). This paper should be discussed in the introduction and cited properly. And any duplication needs to be removed. Somebody who is excellent at writing scientific papers in English should be asked to rewrite the paper. Abstract Palm oil is not necessarily the foremost vegetable oil. For example, soy oil is arguably more important. The abstract is vague. What do you mean by using nucleic acid as a template? The English is poor, e.g. “BSR is characterised by stem decay large-perennial, woody brackets basidiocarps”. What does this mean? The conclusion does not make sense to me at all. Introduction India is only a small palm oil producing country in contradiction to what you have written. You say Africa produces 3% oil palm and then West Africa produces 4%. Which is it? G. boninense is the most serious disease of oil palm in SE Asia and Oceania. In Africa it is considered that fusarium wilt is the most serious fungal disease. G. boninense is accepted as the fungus responsible for BSR in SE Asia at least. You need to mention that G. ryvardenii was identified in Cameroon before and give the citation. Material and Methods Figure 1 should be in the results. In the sampling and isolation section, what does “including fruiting bodies (Basidiocarps) and Ganoderma sp. (Dikaryotic)” mean? How were specimens tested for purity? What does “Microzone, UK, proprietary formulation microLYSIS®-PLUS (MLP) underwent rapid thermal cycling for cell lysis and release of DNA” mean? GanET is G. boninense specific (or better, a palm clade specific) primer. It is not basidiomycete specific. Please correct this. Results Did you test the difference between upper stem rot specimens and basal stem rot specimens? These have been considered as different species before. Phytophthora is not considered to be a fungus. It is an oomycete. The Phylogenetic results section is poor with inadequate information. Discussion Cameroon is not in west Africa. References Lekete-Lawson E, van der Puije GC, Osekre EA and Ackah FK. First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved with reservations]. F1000Research 2025, 14 :413 ( https://doi.org/10.12688/f1000research.161972.1 ) Lekete-Lawson, E, Osekre, E.A. van der Puije, G., Swanzy, et al. 2024. Prevalence of basal stem rot disease caused by Ganoderma sp. on oil palm ( Elaeis guineensis Jacq) in Ghana. Journal of Science and Technology, Special Issue No 1, pp 47 – 61. Competing Interests: None Close Report a concern Discussion is closed on this version, please comment on the latest version above. Author details Author details 1 CSIR-Oil Palm Research Institute, Kade, Eastern Region, Ghana 2 Department of Crop Science, University of Cape Coast College of Agriculture and Natural Sciences, Cape Coast, Ghana 3 Department of Microbiology, Nutrition and Dietetics, Czech University of Life Sciences, Faculty of Agrobiology Food and Natural Resources, Prague, Czech Republic 4 Department of Crop and Soil Sciences, Kwame Nkrumah University of Science and Technology College of Agriculture and Natural Resources, Kumasi, Ghana Emmanuellah Lekete-Lawson Roles: Investigation, Methodology, Resources, Writing – Original Draft Preparation Grace C. van der Puije Roles: Supervision Enoch A. Osekre Roles: Supervision, Writing – Review & Editing Frank K. Ackah Roles: Supervision Competing interests No competing interests were disclosed. Grant information The author(s) declared that no grants were involved in supporting this work. Article Versions (2) version 2 Revised Published: 07 Jul 2025, 14:413 https://doi.org/10.12688/f1000research.161972.2 version 1 Published: 08 Apr 2025, 14:413 https://doi.org/10.12688/f1000research.161972.1 Copyright © 2025 Lekete-Lawson E et al . This is an open access article 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. Download Export To Sciwheel Bibtex EndNote ProCite Ref. Manager (RIS) Sente metrics Views Downloads F1000Research - - PubMed Central info_outline Data from PMC are received and updated monthly. - - Citations open_in_new 0 open_in_new 0 open_in_new SEE MORE DETAILS CITE how to cite this article Lekete-Lawson E, van der Puije GC, Osekre EA and Ackah FK. First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved, 2 approved with reservations] . F1000Research 2025, 14 :413 ( https://doi.org/10.12688/f1000research.161972.1 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS track receive updates on this article Track an article to receive email alerts on any updates to this article. TRACK THIS ARTICLE Share Open Peer Review Current Reviewer Status: ? Key to Reviewer Statuses VIEW HIDE Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Version 1 VERSION 1 PUBLISHED 08 Apr 2025 Views 0 Cite How to cite this report: Lisnawita L. Reviewer Report For: First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved, 2 approved with reservations] . F1000Research 2025, 14 :413 ( https://doi.org/10.5256/f1000research.178088.r380554 ) The direct URL for this report is: https://f1000research.com/articles/14-413/v1#referee-response-380554 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 30 May 2025 Lisnawita Lisnawita , Universitas Sumatera Utara,, Padang Bulan, Indonesia Approved VIEWS 0 https://doi.org/10.5256/f1000research.178088.r380554 In the method, it is not written how many oil palm plants were observed in the field research, but in the result is written 3000 oil palm trees were assessed. Figure 2, 4, 5, 6 are not ... Continue reading READ ALL In the method, it is not written how many oil palm plants were observed in the field research, but in the result is written 3000 oil palm trees were assessed. Figure 2, 4, 5, 6 are not clear Microscopic observations should be accompanied by Ganoderma spore images Researchers only used ITS and GanET for molecular identification. It is recommended to add additional genetic markers such as TEF1α or RPB2 for deeper validation, because the genus Ganoderma is known to have taxonomic complexity Conclusions should be stated explicitly. Do not state that the isolate is “probably” a new species. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests: No competing interests were disclosed. Reviewer Expertise: Phytopathology, Nematology I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Lisnawita L. Reviewer Report For: First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved, 2 approved with reservations] . F1000Research 2025, 14 :413 ( https://doi.org/10.5256/f1000research.178088.r380554 ) The direct URL for this report is: https://f1000research.com/articles/14-413/v1#referee-response-380554 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Author Response 11 Sep 2025 Emmanuellah Lekete-Lawson , Dep. of Microbiology, Czech University of Life Sciences Prague, Prague, Czech Republic 11 Sep 2025 Author Response Reviewer 3: Comment1: In the method, it is not written how many oil palm plants were observed in the field research, but in the result is written 3000 oil palm ... Continue reading Reviewer 3: Comment1: In the method, it is not written how many oil palm plants were observed in the field research, but in the result is written 3000 oil palm trees were assessed. Response1: It was stated in the Materials and methods that 30 oil palm communities/plantation were assessed and 100 palms per plantation, hence the total palms assessed were 3000 palms Comment 2: Figure 2, 4, 5, 6 are not clear Response 2 :6 was deleted the rest have been worked on Comment 3: Microscopic observations should be accompanied by Ganoderma spore images Response 3: Conscious effort was made to get the spore to aid the identification, but proof futile because of constant contamination and other minor Ganoderma species however the project is ongoing and plan to raise the mushroom invitro to reduce contamination is underway. Comment 4: Researchers only used ITS and GanET for molecular identification. It is recommended to add additional genetic markers such as TEF1α or RPB2 for deeper validation, because the genus Ganoderma is known to have taxonomic complexity Response 4: ITS and GanET were primers used by previous researchers, and this is our first time of working on Ganoderma, so we tried to follow the existing protocols/steps of the previous researchers for easy comparison and validation of our results. Comment 5: Conclusions should be stated explicitly. Do not state that the isolate is “probably” a new species. Response 5: Have been corrected. Reviewer 3: Comment1: In the method, it is not written how many oil palm plants were observed in the field research, but in the result is written 3000 oil palm trees were assessed. Response1: It was stated in the Materials and methods that 30 oil palm communities/plantation were assessed and 100 palms per plantation, hence the total palms assessed were 3000 palms Comment 2: Figure 2, 4, 5, 6 are not clear Response 2 :6 was deleted the rest have been worked on Comment 3: Microscopic observations should be accompanied by Ganoderma spore images Response 3: Conscious effort was made to get the spore to aid the identification, but proof futile because of constant contamination and other minor Ganoderma species however the project is ongoing and plan to raise the mushroom invitro to reduce contamination is underway. Comment 4: Researchers only used ITS and GanET for molecular identification. It is recommended to add additional genetic markers such as TEF1α or RPB2 for deeper validation, because the genus Ganoderma is known to have taxonomic complexity Response 4: ITS and GanET were primers used by previous researchers, and this is our first time of working on Ganoderma, so we tried to follow the existing protocols/steps of the previous researchers for easy comparison and validation of our results. Comment 5: Conclusions should be stated explicitly. Do not state that the isolate is “probably” a new species. Response 5: Have been corrected. Competing Interests: Authors declare no competing interest Close Report a concern Respond or Comment COMMENTS ON THIS REPORT Author Response 11 Sep 2025 Emmanuellah Lekete-Lawson , Dep. of Microbiology, Czech University of Life Sciences Prague, Prague, Czech Republic 11 Sep 2025 Author Response Reviewer 3: Comment1: In the method, it is not written how many oil palm plants were observed in the field research, but in the result is written 3000 oil palm ... Continue reading Reviewer 3: Comment1: In the method, it is not written how many oil palm plants were observed in the field research, but in the result is written 3000 oil palm trees were assessed. Response1: It was stated in the Materials and methods that 30 oil palm communities/plantation were assessed and 100 palms per plantation, hence the total palms assessed were 3000 palms Comment 2: Figure 2, 4, 5, 6 are not clear Response 2 :6 was deleted the rest have been worked on Comment 3: Microscopic observations should be accompanied by Ganoderma spore images Response 3: Conscious effort was made to get the spore to aid the identification, but proof futile because of constant contamination and other minor Ganoderma species however the project is ongoing and plan to raise the mushroom invitro to reduce contamination is underway. Comment 4: Researchers only used ITS and GanET for molecular identification. It is recommended to add additional genetic markers such as TEF1α or RPB2 for deeper validation, because the genus Ganoderma is known to have taxonomic complexity Response 4: ITS and GanET were primers used by previous researchers, and this is our first time of working on Ganoderma, so we tried to follow the existing protocols/steps of the previous researchers for easy comparison and validation of our results. Comment 5: Conclusions should be stated explicitly. Do not state that the isolate is “probably” a new species. Response 5: Have been corrected. Reviewer 3: Comment1: In the method, it is not written how many oil palm plants were observed in the field research, but in the result is written 3000 oil palm trees were assessed. Response1: It was stated in the Materials and methods that 30 oil palm communities/plantation were assessed and 100 palms per plantation, hence the total palms assessed were 3000 palms Comment 2: Figure 2, 4, 5, 6 are not clear Response 2 :6 was deleted the rest have been worked on Comment 3: Microscopic observations should be accompanied by Ganoderma spore images Response 3: Conscious effort was made to get the spore to aid the identification, but proof futile because of constant contamination and other minor Ganoderma species however the project is ongoing and plan to raise the mushroom invitro to reduce contamination is underway. Comment 4: Researchers only used ITS and GanET for molecular identification. It is recommended to add additional genetic markers such as TEF1α or RPB2 for deeper validation, because the genus Ganoderma is known to have taxonomic complexity Response 4: ITS and GanET were primers used by previous researchers, and this is our first time of working on Ganoderma, so we tried to follow the existing protocols/steps of the previous researchers for easy comparison and validation of our results. Comment 5: Conclusions should be stated explicitly. Do not state that the isolate is “probably” a new species. Response 5: Have been corrected. Competing Interests: Authors declare no competing interest Close Report a concern COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Tondok ET. Reviewer Report For: First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved, 2 approved with reservations] . F1000Research 2025, 14 :413 ( https://doi.org/10.5256/f1000research.178088.r380559 ) The direct URL for this report is: https://f1000research.com/articles/14-413/v1#referee-response-380559 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 19 May 2025 Efi Toding Tondok , Plant Protection, IPB University, Bogor, West Java, Indonesia Approved with Reservations VIEWS 0 https://doi.org/10.5256/f1000research.178088.r380559 Method . 1)Scores on disease severity must be written clearly. 2) This is Basidiomycota, so you will never find spores by mounting mycelia on object glass. Result . 1) Please add basidiospores morphometrics as one of the important ... Continue reading READ ALL Method . 1)Scores on disease severity must be written clearly. 2) This is Basidiomycota, so you will never find spores by mounting mycelia on object glass. Result . 1) Please add basidiospores morphometrics as one of the important features for morphological identification. The 0.5 mm bar is too long for microscopic measurement. 2) Fig 5 and 6: please put the objects in a good arrangement, so the reader will be easy to understand. Don't forget the control. The 0.5 mm bar is very annoying. 3) It is very important to include pictures from the fields, the mild or early symptom to severe or advance symptom. Formation of basidiocarp indicating the advance symptom, plant almost die. 4) How many species of Ganoderma found in Ghana? 5) Please put the same codes on your isolates since morphological identification - pathogenicity tests - molecular identification: easy to track it, to prove that the tested isolate is the same as the isolate identified morphologically and molecularly Discussion. Please explain why disease incidence and disease severity are highest in K29-B, compared to other places Conclusion. To prove that the primers you use can distinguish Ganoderma well, you need to test them on several Ganoderma species of known species. So, please go only to your finding. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? Partly Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Partly Competing Interests: No competing interests were disclosed. Reviewer Expertise: Phytopathology, Mycology, Seed Pathology, Biological Control I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Tondok ET. Reviewer Report For: First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved, 2 approved with reservations] . F1000Research 2025, 14 :413 ( https://doi.org/10.5256/f1000research.178088.r380559 ) The direct URL for this report is: https://f1000research.com/articles/14-413/v1#referee-response-380559 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Author Response 11 Sep 2025 Emmanuellah Lekete-Lawson , Dep. of Microbiology, Czech University of Life Sciences Prague, Prague, Czech Republic 11 Sep 2025 Author Response Response to reviewer’s comment 2- Reviewer: Robert Paterson, Centre for Biological Engineering, University of Minho, Braga, Portugal Comment on Lekete-Lawson et al. (2025) By: Robert Russell Monteith Paterson University of ... Continue reading Response to reviewer’s comment 2- Reviewer: Robert Paterson, Centre for Biological Engineering, University of Minho, Braga, Portugal Comment on Lekete-Lawson et al. (2025) By: Robert Russell Monteith Paterson University of Minho, Portugal [email protected] #My first concern when reading Lekete-Lawson et al. (2025) was the PCR analysis and subsequently I detected other problems. PCR Comment1: In relation to the PCR, in the text it says Figure 7A is for the culture based samples but these are Figure 7B in the legend to the image of the gel. Figure 7 B in the text is for basidiocarps but these are 7A in the legend to the figure. Which is it? Response:1 It was a typing mistake and has been corrected see page 14, Figure 7. Comment 2. Also, there is no information about the strain numbers that represented tracks 1 to 24 in the figure of the gels. These should be provided. It is all very confused. Also, there appeared to be a faint band in the blanks for samples 1 to 24 which were at the same intensity for track 20. Presumably the blanks were contaminated. Were tracks 20 and 35 from the same specimen and strain? Response 2. Yes, they are from the same sample but cannot confirm the strain, identity until confirm molecularly, this is because we were dealing with several new strains the faintness could probably due to either contamination from the DNA extraction or poor gel loading or not what we were expecting. b . The strain number could be captured in the gel because the software we were using was not programmed in that order, however detail of the individual strain numbers have been provided in the datataset repository. Comment 3: Was there a procedure for identifying inhibition of the PCR? I know that an internal amplification control (IAC) to assess inhibition is recommended for at least some PCRs. IAC would provide information on inhibition or could it be that the samples represented by 20 and 35 were not what you thought they were? Response.3a. We first suspected inhibitors like EDTA/Salts or even phenol after thoroughly purification DNA products; we still had the same results, probably not what we were expecting. Why did you not run known relevant Ganoderma species for comparison with the other specimens? This would provide information on the novelty of your specimens. Overall, there are considerable problems with the presentation of the PCR results. Response 3b . We were running what was available, we tried to identify and confirm if what we have in our country could be the same elsewhere. Having gotten the sequence data (Accession numbers) we then compared them with the known and existing ones in the data base, which helped us confirm our findings. Other concerns are as follows: General A strain and specimen list is necessary with accession numbers. Response: Strain and accession numbers have been provided in the write-up, together with phylogenetic trees giving a detailed explanation. Moreover, other details explanations/documents can be found in the figshare repository Comment 4 Are you saying G. ryvardenii is the only species involved in BSR in Ghana? What is the new species you say you have discovered? This is unclear. Response 4: Please I have stated it with percentage occurrence of the Ryvadenii and other species identified, however, The occurrence of G. ryvadenii was higher 83% compared to the remaining ones identified, even there was 1% G. woerese which has never been identified anywhere hence was discarded. So in total four different Ganoderma species were identified including the famous G. boninnense . But there percentages were very minimal. Comment: A considerable amount of this paper is taken from a previous paper (Lekete-Lawson et al. 2024). This paper should be discussed in the introduction and cited properly. And any duplication needs to be removed. Response: The study is ongoing with three main objectives, each autonomous but with a common goal of solving a disease problem. For each objective achieved, we release a publication. There was no repetition of the writing; rather, we showed how one objective is linked to another by adopting some of the procedures used in a previous to achieve the current objective and referencing it appropriately, it is not repetition but continuation and updates. Comment: Somebody who is excellent at writing scientific papers in English should be asked to rewrite the paper. Response: Well noted Abstract Comment: Palm oil is not necessarily the foremost vegetable oil. For example, soy oil is arguably more important. Response: This is very true, but Several studies (Breton et al, 2006; Bharudan et al., 2022) have proven that oil palm is a naturally and high-yielding oil crop,13 times more than soy and any other oil crops combined. Oil palm does not need any artificial additives to generate its oil. Comment: The abstract is vague??? What do you mean by using nucleic acid as a template? Response: If we said we used nucleic acid as a template, it means we used DNA as the starting material in a molecular assay to amplify the genetic sequences. For example, we extracted DNA from the culture-based isolates and that of the Basidiocarps and in this context, nucleic acid as a template means DNA was used as an input material to generate measurable results in the experiment. Comment: The English is poor, e.g. “BSR is characterised by stem decay, large-perennial, woody brackets basidiocarps”. What does this mean? Response: This is rewritten as: “Basal Stem Rot (BSR) is characterised by progressive stem decay coupled with the formation of large, perennial, woody basidiocarps with the average measurement of 2-65 cm in diameter on infected palms”. Comment: The conclusion does not make sense to me at all. Response: Conclusion has been rewritten “This study presents the first report of Ganoderma ryvardenii causing BSR disease on oil palm in Ghana, potentially the first in West Africa, and second in Africa. Notably, the pathogen was previously first reported to cause similar disease on oil palm in Cameroon, highlighting its emerging threat to oil palm production in the Sub-Saharan African region”(See page 2 of the manuscript) Introduction India is only a small palm oil-producing country in contradiction to what you have written. Respponse: Okay, well noted. Comment: You say Africa produces 3% oil palm, and then West Africa produces 4%. Which is it? Response: Africa accounts for approximately 3% of global palm oil production, with West Africa contributing about 4% of the continent’s total output. This implies that West Africa contributes 4% of Africa’s 3% share of global production. For example, if Africa produces 100,000 metric tonnes of palm oil, representing 3% of the global total, then West Africa contributes approximately 4,000 metric tonnes." Comment: G. boninense is the most serious disease of oil palm in SE Asia and Oceania. In Africa, it is considered that Fusarium wilt is the most serious fungal disease. Response: Contrary to your assertion in Ghana, research has shown that Fusarium is no longer a serious disease in our plantation and certainly not Ganoderma too. the case about Ganoderma is an emergent one, though reported many decades ago by Turner (1967), but was regarded as not economically important till now. Comment: G. boninense is accepted as the fungus responsible for BSR in SE Asia at least. You need to mention that G. ryvardenii was identified in Cameroon before and give the citation. Response: This was corrected in the introduction (See page 3) Materials and Methods Figure 1 should be in the results. Response: That was how it was structured, in the original manuscript, but changed by the editor Comment: In the sampling and isolation section, what does “including fruiting bodies (Basidiocarps) and Ganoderma sp. (Dikaryotic)” mean? Response: This was rewritten for better clarity as: “A total of ninety (90) symptomatic samples comprising basidiocarps and infected rotten plant tissues were collected from oil palms across the three agro-ecological regions under investigation. Comment: How were specimens tested for purity? Response: This was done with the combination of visual inspection and DNA-based confirmation (advanced purity check) using Universal (ITS) primers, thus a single clear band is likely considered pure. Comment: What does “Microzone, UK, proprietary formulation microLYSIS®-PLUS (MLP) underwent rapid thermal cycling for cell lysis and release of DNA” mean? Response: For DNA extraction, a proprietary formulation, microLYSIS ® -PLUS (MLP) from Microzone, UK, was used, for rapid thermal cycling to ensure efficient cell lysis and DNA release. Comment: GanET is G. boninense specific (or better, a palm clade specific) primer. It is not basidiomycete-specific. Please correct this. Response: Well noted, however, according to Mei Lieng Lo (2023) and Mandal et al., (2014), GanET is a DNA primer specifically designed for detection of Ganoderma boninense from oil palm tissue. Their study highlights the effectiveness of this primer in early pathogen detection even before visible symptoms appear. Results Comment: Did you test the difference between upper stem rot specimens and basal stem rot specimens? These have been considered as different species before. Response: Yes, we did, and it is still under investigation. This is because we identified it on only one palm at one spot, so we put the entire plantation and the area under surveillance, looking for more evidence to justify its discovery. Upper Stem Rot is known to be caused by Ganoderma Zonatum , which is related to G. boninense . In our study, we did not identify G. zonatum, but rather we had 7 % matching G. wiiroense and 4 % matches G. boninense. Nonetheless, G. wiiroense appeared to be a new strain/ species which has never been identified or published before, hence placed under further investigation. Comment: Phytophthora is not considered to be a fungus. It is an oomycete. Response: Well noted The Phylogenetic results section is poor with inadequate information. Response : More detailed explanation has been added. Additional data were provided in the figshare repository. (Figshare repository and can be accessed at [DOI: 10.6084/m9.figshare.29382632). Discussion Comment: Cameroon is not in West Africa. Response: Cameroon is in Central Africa, not in West Africa, but it is part of Sub-Saharan African countries, including Ghana. Response to reviewer’s comment 2- Reviewer: Robert Paterson, Centre for Biological Engineering, University of Minho, Braga, Portugal Comment on Lekete-Lawson et al. (2025) By: Robert Russell Monteith Paterson University of Minho, Portugal [email protected] #My first concern when reading Lekete-Lawson et al. (2025) was the PCR analysis and subsequently I detected other problems. PCR Comment1: In relation to the PCR, in the text it says Figure 7A is for the culture based samples but these are Figure 7B in the legend to the image of the gel. Figure 7 B in the text is for basidiocarps but these are 7A in the legend to the figure. Which is it? Response:1 It was a typing mistake and has been corrected see page 14, Figure 7. Comment 2. Also, there is no information about the strain numbers that represented tracks 1 to 24 in the figure of the gels. These should be provided. It is all very confused. Also, there appeared to be a faint band in the blanks for samples 1 to 24 which were at the same intensity for track 20. Presumably the blanks were contaminated. Were tracks 20 and 35 from the same specimen and strain? Response 2. Yes, they are from the same sample but cannot confirm the strain, identity until confirm molecularly, this is because we were dealing with several new strains the faintness could probably due to either contamination from the DNA extraction or poor gel loading or not what we were expecting. b . The strain number could be captured in the gel because the software we were using was not programmed in that order, however detail of the individual strain numbers have been provided in the datataset repository. Comment 3: Was there a procedure for identifying inhibition of the PCR? I know that an internal amplification control (IAC) to assess inhibition is recommended for at least some PCRs. IAC would provide information on inhibition or could it be that the samples represented by 20 and 35 were not what you thought they were? Response.3a. We first suspected inhibitors like EDTA/Salts or even phenol after thoroughly purification DNA products; we still had the same results, probably not what we were expecting. Why did you not run known relevant Ganoderma species for comparison with the other specimens? This would provide information on the novelty of your specimens. Overall, there are considerable problems with the presentation of the PCR results. Response 3b . We were running what was available, we tried to identify and confirm if what we have in our country could be the same elsewhere. Having gotten the sequence data (Accession numbers) we then compared them with the known and existing ones in the data base, which helped us confirm our findings. Other concerns are as follows: General A strain and specimen list is necessary with accession numbers. Response: Strain and accession numbers have been provided in the write-up, together with phylogenetic trees giving a detailed explanation. Moreover, other details explanations/documents can be found in the figshare repository Comment 4 Are you saying G. ryvardenii is the only species involved in BSR in Ghana? What is the new species you say you have discovered? This is unclear. Response 4: Please I have stated it with percentage occurrence of the Ryvadenii and other species identified, however, The occurrence of G. ryvadenii was higher 83% compared to the remaining ones identified, even there was 1% G. woerese which has never been identified anywhere hence was discarded. So in total four different Ganoderma species were identified including the famous G. boninnense . But there percentages were very minimal. Comment: A considerable amount of this paper is taken from a previous paper (Lekete-Lawson et al. 2024). This paper should be discussed in the introduction and cited properly. And any duplication needs to be removed. Response: The study is ongoing with three main objectives, each autonomous but with a common goal of solving a disease problem. For each objective achieved, we release a publication. There was no repetition of the writing; rather, we showed how one objective is linked to another by adopting some of the procedures used in a previous to achieve the current objective and referencing it appropriately, it is not repetition but continuation and updates. Comment: Somebody who is excellent at writing scientific papers in English should be asked to rewrite the paper. Response: Well noted Abstract Comment: Palm oil is not necessarily the foremost vegetable oil. For example, soy oil is arguably more important. Response: This is very true, but Several studies (Breton et al, 2006; Bharudan et al., 2022) have proven that oil palm is a naturally and high-yielding oil crop,13 times more than soy and any other oil crops combined. Oil palm does not need any artificial additives to generate its oil. Comment: The abstract is vague??? What do you mean by using nucleic acid as a template? Response: If we said we used nucleic acid as a template, it means we used DNA as the starting material in a molecular assay to amplify the genetic sequences. For example, we extracted DNA from the culture-based isolates and that of the Basidiocarps and in this context, nucleic acid as a template means DNA was used as an input material to generate measurable results in the experiment. Comment: The English is poor, e.g. “BSR is characterised by stem decay, large-perennial, woody brackets basidiocarps”. What does this mean? Response: This is rewritten as: “Basal Stem Rot (BSR) is characterised by progressive stem decay coupled with the formation of large, perennial, woody basidiocarps with the average measurement of 2-65 cm in diameter on infected palms”. Comment: The conclusion does not make sense to me at all. Response: Conclusion has been rewritten “This study presents the first report of Ganoderma ryvardenii causing BSR disease on oil palm in Ghana, potentially the first in West Africa, and second in Africa. Notably, the pathogen was previously first reported to cause similar disease on oil palm in Cameroon, highlighting its emerging threat to oil palm production in the Sub-Saharan African region”(See page 2 of the manuscript) Introduction India is only a small palm oil-producing country in contradiction to what you have written. Respponse: Okay, well noted. Comment: You say Africa produces 3% oil palm, and then West Africa produces 4%. Which is it? Response: Africa accounts for approximately 3% of global palm oil production, with West Africa contributing about 4% of the continent’s total output. This implies that West Africa contributes 4% of Africa’s 3% share of global production. For example, if Africa produces 100,000 metric tonnes of palm oil, representing 3% of the global total, then West Africa contributes approximately 4,000 metric tonnes." Comment: G. boninense is the most serious disease of oil palm in SE Asia and Oceania. In Africa, it is considered that Fusarium wilt is the most serious fungal disease. Response: Contrary to your assertion in Ghana, research has shown that Fusarium is no longer a serious disease in our plantation and certainly not Ganoderma too. the case about Ganoderma is an emergent one, though reported many decades ago by Turner (1967), but was regarded as not economically important till now. Comment: G. boninense is accepted as the fungus responsible for BSR in SE Asia at least. You need to mention that G. ryvardenii was identified in Cameroon before and give the citation. Response: This was corrected in the introduction (See page 3) Materials and Methods Figure 1 should be in the results. Response: That was how it was structured, in the original manuscript, but changed by the editor Comment: In the sampling and isolation section, what does “including fruiting bodies (Basidiocarps) and Ganoderma sp. (Dikaryotic)” mean? Response: This was rewritten for better clarity as: “A total of ninety (90) symptomatic samples comprising basidiocarps and infected rotten plant tissues were collected from oil palms across the three agro-ecological regions under investigation. Comment: How were specimens tested for purity? Response: This was done with the combination of visual inspection and DNA-based confirmation (advanced purity check) using Universal (ITS) primers, thus a single clear band is likely considered pure. Comment: What does “Microzone, UK, proprietary formulation microLYSIS®-PLUS (MLP) underwent rapid thermal cycling for cell lysis and release of DNA” mean? Response: For DNA extraction, a proprietary formulation, microLYSIS ® -PLUS (MLP) from Microzone, UK, was used, for rapid thermal cycling to ensure efficient cell lysis and DNA release. Comment: GanET is G. boninense specific (or better, a palm clade specific) primer. It is not basidiomycete-specific. Please correct this. Response: Well noted, however, according to Mei Lieng Lo (2023) and Mandal et al., (2014), GanET is a DNA primer specifically designed for detection of Ganoderma boninense from oil palm tissue. Their study highlights the effectiveness of this primer in early pathogen detection even before visible symptoms appear. Results Comment: Did you test the difference between upper stem rot specimens and basal stem rot specimens? These have been considered as different species before. Response: Yes, we did, and it is still under investigation. This is because we identified it on only one palm at one spot, so we put the entire plantation and the area under surveillance, looking for more evidence to justify its discovery. Upper Stem Rot is known to be caused by Ganoderma Zonatum , which is related to G. boninense . In our study, we did not identify G. zonatum, but rather we had 7 % matching G. wiiroense and 4 % matches G. boninense. Nonetheless, G. wiiroense appeared to be a new strain/ species which has never been identified or published before, hence placed under further investigation. Comment: Phytophthora is not considered to be a fungus. It is an oomycete. Response: Well noted The Phylogenetic results section is poor with inadequate information. Response : More detailed explanation has been added. Additional data were provided in the figshare repository. (Figshare repository and can be accessed at [DOI: 10.6084/m9.figshare.29382632). Discussion Comment: Cameroon is not in West Africa. Response: Cameroon is in Central Africa, not in West Africa, but it is part of Sub-Saharan African countries, including Ghana. Competing Interests: No competing interest Close Report a concern Respond or Comment COMMENTS ON THIS REPORT Author Response 11 Sep 2025 Emmanuellah Lekete-Lawson , Dep. of Microbiology, Czech University of Life Sciences Prague, Prague, Czech Republic 11 Sep 2025 Author Response Response to reviewer’s comment 2- Reviewer: Robert Paterson, Centre for Biological Engineering, University of Minho, Braga, Portugal Comment on Lekete-Lawson et al. (2025) By: Robert Russell Monteith Paterson University of ... Continue reading Response to reviewer’s comment 2- Reviewer: Robert Paterson, Centre for Biological Engineering, University of Minho, Braga, Portugal Comment on Lekete-Lawson et al. (2025) By: Robert Russell Monteith Paterson University of Minho, Portugal [email protected] #My first concern when reading Lekete-Lawson et al. (2025) was the PCR analysis and subsequently I detected other problems. PCR Comment1: In relation to the PCR, in the text it says Figure 7A is for the culture based samples but these are Figure 7B in the legend to the image of the gel. Figure 7 B in the text is for basidiocarps but these are 7A in the legend to the figure. Which is it? Response:1 It was a typing mistake and has been corrected see page 14, Figure 7. Comment 2. Also, there is no information about the strain numbers that represented tracks 1 to 24 in the figure of the gels. These should be provided. It is all very confused. Also, there appeared to be a faint band in the blanks for samples 1 to 24 which were at the same intensity for track 20. Presumably the blanks were contaminated. Were tracks 20 and 35 from the same specimen and strain? Response 2. Yes, they are from the same sample but cannot confirm the strain, identity until confirm molecularly, this is because we were dealing with several new strains the faintness could probably due to either contamination from the DNA extraction or poor gel loading or not what we were expecting. b . The strain number could be captured in the gel because the software we were using was not programmed in that order, however detail of the individual strain numbers have been provided in the datataset repository. Comment 3: Was there a procedure for identifying inhibition of the PCR? I know that an internal amplification control (IAC) to assess inhibition is recommended for at least some PCRs. IAC would provide information on inhibition or could it be that the samples represented by 20 and 35 were not what you thought they were? Response.3a. We first suspected inhibitors like EDTA/Salts or even phenol after thoroughly purification DNA products; we still had the same results, probably not what we were expecting. Why did you not run known relevant Ganoderma species for comparison with the other specimens? This would provide information on the novelty of your specimens. Overall, there are considerable problems with the presentation of the PCR results. Response 3b . We were running what was available, we tried to identify and confirm if what we have in our country could be the same elsewhere. Having gotten the sequence data (Accession numbers) we then compared them with the known and existing ones in the data base, which helped us confirm our findings. Other concerns are as follows: General A strain and specimen list is necessary with accession numbers. Response: Strain and accession numbers have been provided in the write-up, together with phylogenetic trees giving a detailed explanation. Moreover, other details explanations/documents can be found in the figshare repository Comment 4 Are you saying G. ryvardenii is the only species involved in BSR in Ghana? What is the new species you say you have discovered? This is unclear. Response 4: Please I have stated it with percentage occurrence of the Ryvadenii and other species identified, however, The occurrence of G. ryvadenii was higher 83% compared to the remaining ones identified, even there was 1% G. woerese which has never been identified anywhere hence was discarded. So in total four different Ganoderma species were identified including the famous G. boninnense . But there percentages were very minimal. Comment: A considerable amount of this paper is taken from a previous paper (Lekete-Lawson et al. 2024). This paper should be discussed in the introduction and cited properly. And any duplication needs to be removed. Response: The study is ongoing with three main objectives, each autonomous but with a common goal of solving a disease problem. For each objective achieved, we release a publication. There was no repetition of the writing; rather, we showed how one objective is linked to another by adopting some of the procedures used in a previous to achieve the current objective and referencing it appropriately, it is not repetition but continuation and updates. Comment: Somebody who is excellent at writing scientific papers in English should be asked to rewrite the paper. Response: Well noted Abstract Comment: Palm oil is not necessarily the foremost vegetable oil. For example, soy oil is arguably more important. Response: This is very true, but Several studies (Breton et al, 2006; Bharudan et al., 2022) have proven that oil palm is a naturally and high-yielding oil crop,13 times more than soy and any other oil crops combined. Oil palm does not need any artificial additives to generate its oil. Comment: The abstract is vague??? What do you mean by using nucleic acid as a template? Response: If we said we used nucleic acid as a template, it means we used DNA as the starting material in a molecular assay to amplify the genetic sequences. For example, we extracted DNA from the culture-based isolates and that of the Basidiocarps and in this context, nucleic acid as a template means DNA was used as an input material to generate measurable results in the experiment. Comment: The English is poor, e.g. “BSR is characterised by stem decay, large-perennial, woody brackets basidiocarps”. What does this mean? Response: This is rewritten as: “Basal Stem Rot (BSR) is characterised by progressive stem decay coupled with the formation of large, perennial, woody basidiocarps with the average measurement of 2-65 cm in diameter on infected palms”. Comment: The conclusion does not make sense to me at all. Response: Conclusion has been rewritten “This study presents the first report of Ganoderma ryvardenii causing BSR disease on oil palm in Ghana, potentially the first in West Africa, and second in Africa. Notably, the pathogen was previously first reported to cause similar disease on oil palm in Cameroon, highlighting its emerging threat to oil palm production in the Sub-Saharan African region”(See page 2 of the manuscript) Introduction India is only a small palm oil-producing country in contradiction to what you have written. Respponse: Okay, well noted. Comment: You say Africa produces 3% oil palm, and then West Africa produces 4%. Which is it? Response: Africa accounts for approximately 3% of global palm oil production, with West Africa contributing about 4% of the continent’s total output. This implies that West Africa contributes 4% of Africa’s 3% share of global production. For example, if Africa produces 100,000 metric tonnes of palm oil, representing 3% of the global total, then West Africa contributes approximately 4,000 metric tonnes." Comment: G. boninense is the most serious disease of oil palm in SE Asia and Oceania. In Africa, it is considered that Fusarium wilt is the most serious fungal disease. Response: Contrary to your assertion in Ghana, research has shown that Fusarium is no longer a serious disease in our plantation and certainly not Ganoderma too. the case about Ganoderma is an emergent one, though reported many decades ago by Turner (1967), but was regarded as not economically important till now. Comment: G. boninense is accepted as the fungus responsible for BSR in SE Asia at least. You need to mention that G. ryvardenii was identified in Cameroon before and give the citation. Response: This was corrected in the introduction (See page 3) Materials and Methods Figure 1 should be in the results. Response: That was how it was structured, in the original manuscript, but changed by the editor Comment: In the sampling and isolation section, what does “including fruiting bodies (Basidiocarps) and Ganoderma sp. (Dikaryotic)” mean? Response: This was rewritten for better clarity as: “A total of ninety (90) symptomatic samples comprising basidiocarps and infected rotten plant tissues were collected from oil palms across the three agro-ecological regions under investigation. Comment: How were specimens tested for purity? Response: This was done with the combination of visual inspection and DNA-based confirmation (advanced purity check) using Universal (ITS) primers, thus a single clear band is likely considered pure. Comment: What does “Microzone, UK, proprietary formulation microLYSIS®-PLUS (MLP) underwent rapid thermal cycling for cell lysis and release of DNA” mean? Response: For DNA extraction, a proprietary formulation, microLYSIS ® -PLUS (MLP) from Microzone, UK, was used, for rapid thermal cycling to ensure efficient cell lysis and DNA release. Comment: GanET is G. boninense specific (or better, a palm clade specific) primer. It is not basidiomycete-specific. Please correct this. Response: Well noted, however, according to Mei Lieng Lo (2023) and Mandal et al., (2014), GanET is a DNA primer specifically designed for detection of Ganoderma boninense from oil palm tissue. Their study highlights the effectiveness of this primer in early pathogen detection even before visible symptoms appear. Results Comment: Did you test the difference between upper stem rot specimens and basal stem rot specimens? These have been considered as different species before. Response: Yes, we did, and it is still under investigation. This is because we identified it on only one palm at one spot, so we put the entire plantation and the area under surveillance, looking for more evidence to justify its discovery. Upper Stem Rot is known to be caused by Ganoderma Zonatum , which is related to G. boninense . In our study, we did not identify G. zonatum, but rather we had 7 % matching G. wiiroense and 4 % matches G. boninense. Nonetheless, G. wiiroense appeared to be a new strain/ species which has never been identified or published before, hence placed under further investigation. Comment: Phytophthora is not considered to be a fungus. It is an oomycete. Response: Well noted The Phylogenetic results section is poor with inadequate information. Response : More detailed explanation has been added. Additional data were provided in the figshare repository. (Figshare repository and can be accessed at [DOI: 10.6084/m9.figshare.29382632). Discussion Comment: Cameroon is not in West Africa. Response: Cameroon is in Central Africa, not in West Africa, but it is part of Sub-Saharan African countries, including Ghana. Response to reviewer’s comment 2- Reviewer: Robert Paterson, Centre for Biological Engineering, University of Minho, Braga, Portugal Comment on Lekete-Lawson et al. (2025) By: Robert Russell Monteith Paterson University of Minho, Portugal [email protected] #My first concern when reading Lekete-Lawson et al. (2025) was the PCR analysis and subsequently I detected other problems. PCR Comment1: In relation to the PCR, in the text it says Figure 7A is for the culture based samples but these are Figure 7B in the legend to the image of the gel. Figure 7 B in the text is for basidiocarps but these are 7A in the legend to the figure. Which is it? Response:1 It was a typing mistake and has been corrected see page 14, Figure 7. Comment 2. Also, there is no information about the strain numbers that represented tracks 1 to 24 in the figure of the gels. These should be provided. It is all very confused. Also, there appeared to be a faint band in the blanks for samples 1 to 24 which were at the same intensity for track 20. Presumably the blanks were contaminated. Were tracks 20 and 35 from the same specimen and strain? Response 2. Yes, they are from the same sample but cannot confirm the strain, identity until confirm molecularly, this is because we were dealing with several new strains the faintness could probably due to either contamination from the DNA extraction or poor gel loading or not what we were expecting. b . The strain number could be captured in the gel because the software we were using was not programmed in that order, however detail of the individual strain numbers have been provided in the datataset repository. Comment 3: Was there a procedure for identifying inhibition of the PCR? I know that an internal amplification control (IAC) to assess inhibition is recommended for at least some PCRs. IAC would provide information on inhibition or could it be that the samples represented by 20 and 35 were not what you thought they were? Response.3a. We first suspected inhibitors like EDTA/Salts or even phenol after thoroughly purification DNA products; we still had the same results, probably not what we were expecting. Why did you not run known relevant Ganoderma species for comparison with the other specimens? This would provide information on the novelty of your specimens. Overall, there are considerable problems with the presentation of the PCR results. Response 3b . We were running what was available, we tried to identify and confirm if what we have in our country could be the same elsewhere. Having gotten the sequence data (Accession numbers) we then compared them with the known and existing ones in the data base, which helped us confirm our findings. Other concerns are as follows: General A strain and specimen list is necessary with accession numbers. Response: Strain and accession numbers have been provided in the write-up, together with phylogenetic trees giving a detailed explanation. Moreover, other details explanations/documents can be found in the figshare repository Comment 4 Are you saying G. ryvardenii is the only species involved in BSR in Ghana? What is the new species you say you have discovered? This is unclear. Response 4: Please I have stated it with percentage occurrence of the Ryvadenii and other species identified, however, The occurrence of G. ryvadenii was higher 83% compared to the remaining ones identified, even there was 1% G. woerese which has never been identified anywhere hence was discarded. So in total four different Ganoderma species were identified including the famous G. boninnense . But there percentages were very minimal. Comment: A considerable amount of this paper is taken from a previous paper (Lekete-Lawson et al. 2024). This paper should be discussed in the introduction and cited properly. And any duplication needs to be removed. Response: The study is ongoing with three main objectives, each autonomous but with a common goal of solving a disease problem. For each objective achieved, we release a publication. There was no repetition of the writing; rather, we showed how one objective is linked to another by adopting some of the procedures used in a previous to achieve the current objective and referencing it appropriately, it is not repetition but continuation and updates. Comment: Somebody who is excellent at writing scientific papers in English should be asked to rewrite the paper. Response: Well noted Abstract Comment: Palm oil is not necessarily the foremost vegetable oil. For example, soy oil is arguably more important. Response: This is very true, but Several studies (Breton et al, 2006; Bharudan et al., 2022) have proven that oil palm is a naturally and high-yielding oil crop,13 times more than soy and any other oil crops combined. Oil palm does not need any artificial additives to generate its oil. Comment: The abstract is vague??? What do you mean by using nucleic acid as a template? Response: If we said we used nucleic acid as a template, it means we used DNA as the starting material in a molecular assay to amplify the genetic sequences. For example, we extracted DNA from the culture-based isolates and that of the Basidiocarps and in this context, nucleic acid as a template means DNA was used as an input material to generate measurable results in the experiment. Comment: The English is poor, e.g. “BSR is characterised by stem decay, large-perennial, woody brackets basidiocarps”. What does this mean? Response: This is rewritten as: “Basal Stem Rot (BSR) is characterised by progressive stem decay coupled with the formation of large, perennial, woody basidiocarps with the average measurement of 2-65 cm in diameter on infected palms”. Comment: The conclusion does not make sense to me at all. Response: Conclusion has been rewritten “This study presents the first report of Ganoderma ryvardenii causing BSR disease on oil palm in Ghana, potentially the first in West Africa, and second in Africa. Notably, the pathogen was previously first reported to cause similar disease on oil palm in Cameroon, highlighting its emerging threat to oil palm production in the Sub-Saharan African region”(See page 2 of the manuscript) Introduction India is only a small palm oil-producing country in contradiction to what you have written. Respponse: Okay, well noted. Comment: You say Africa produces 3% oil palm, and then West Africa produces 4%. Which is it? Response: Africa accounts for approximately 3% of global palm oil production, with West Africa contributing about 4% of the continent’s total output. This implies that West Africa contributes 4% of Africa’s 3% share of global production. For example, if Africa produces 100,000 metric tonnes of palm oil, representing 3% of the global total, then West Africa contributes approximately 4,000 metric tonnes." Comment: G. boninense is the most serious disease of oil palm in SE Asia and Oceania. In Africa, it is considered that Fusarium wilt is the most serious fungal disease. Response: Contrary to your assertion in Ghana, research has shown that Fusarium is no longer a serious disease in our plantation and certainly not Ganoderma too. the case about Ganoderma is an emergent one, though reported many decades ago by Turner (1967), but was regarded as not economically important till now. Comment: G. boninense is accepted as the fungus responsible for BSR in SE Asia at least. You need to mention that G. ryvardenii was identified in Cameroon before and give the citation. Response: This was corrected in the introduction (See page 3) Materials and Methods Figure 1 should be in the results. Response: That was how it was structured, in the original manuscript, but changed by the editor Comment: In the sampling and isolation section, what does “including fruiting bodies (Basidiocarps) and Ganoderma sp. (Dikaryotic)” mean? Response: This was rewritten for better clarity as: “A total of ninety (90) symptomatic samples comprising basidiocarps and infected rotten plant tissues were collected from oil palms across the three agro-ecological regions under investigation. Comment: How were specimens tested for purity? Response: This was done with the combination of visual inspection and DNA-based confirmation (advanced purity check) using Universal (ITS) primers, thus a single clear band is likely considered pure. Comment: What does “Microzone, UK, proprietary formulation microLYSIS®-PLUS (MLP) underwent rapid thermal cycling for cell lysis and release of DNA” mean? Response: For DNA extraction, a proprietary formulation, microLYSIS ® -PLUS (MLP) from Microzone, UK, was used, for rapid thermal cycling to ensure efficient cell lysis and DNA release. Comment: GanET is G. boninense specific (or better, a palm clade specific) primer. It is not basidiomycete-specific. Please correct this. Response: Well noted, however, according to Mei Lieng Lo (2023) and Mandal et al., (2014), GanET is a DNA primer specifically designed for detection of Ganoderma boninense from oil palm tissue. Their study highlights the effectiveness of this primer in early pathogen detection even before visible symptoms appear. Results Comment: Did you test the difference between upper stem rot specimens and basal stem rot specimens? These have been considered as different species before. Response: Yes, we did, and it is still under investigation. This is because we identified it on only one palm at one spot, so we put the entire plantation and the area under surveillance, looking for more evidence to justify its discovery. Upper Stem Rot is known to be caused by Ganoderma Zonatum , which is related to G. boninense . In our study, we did not identify G. zonatum, but rather we had 7 % matching G. wiiroense and 4 % matches G. boninense. Nonetheless, G. wiiroense appeared to be a new strain/ species which has never been identified or published before, hence placed under further investigation. Comment: Phytophthora is not considered to be a fungus. It is an oomycete. Response: Well noted The Phylogenetic results section is poor with inadequate information. Response : More detailed explanation has been added. Additional data were provided in the figshare repository. (Figshare repository and can be accessed at [DOI: 10.6084/m9.figshare.29382632). Discussion Comment: Cameroon is not in West Africa. Response: Cameroon is in Central Africa, not in West Africa, but it is part of Sub-Saharan African countries, including Ghana. Competing Interests: No competing interest Close Report a concern COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Supriyanto S. Reviewer Report For: First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved, 2 approved with reservations] . F1000Research 2025, 14 :413 ( https://doi.org/10.5256/f1000research.178088.r377597 ) The direct URL for this report is: https://f1000research.com/articles/14-413/v1#referee-response-377597 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 30 Apr 2025 Supriyanto Supriyanto , Tanjungpura University, Jl. Prof. Dr. H. Hadari Nawawi, Pontianak, Indonesia Approved with Reservations VIEWS 0 https://doi.org/10.5256/f1000research.178088.r377597 Overall, this article is well written. However, some parts need to be strengthened to make it a reliable reference. 1. In the last paragraph of the introduction, considering the author's claim about the discovery of a new species of ... Continue reading READ ALL Overall, this article is well written. However, some parts need to be strengthened to make it a reliable reference. 1. In the last paragraph of the introduction, considering the author's claim about the discovery of a new species of Ganoderma that causes Basal Stem Rot disease in oil palm, it is better to add a new, more complete reference about reports related to the types of Ganoderma that previous researchers have reported. 2. Method. Observation of disease severity in oil palm in the field uses a score of 0 to 4. The scoring criteria should be displayed in full. In addition, the use of the number of fruiting bodies as an indicator of severity seems rather risky because there has been no report on this. Based on experience in Indonesia, the number of fruiting bodies that can be found is not directly related to the severity of the disease. 3. Method, pathogenicity test. To be clearer, it is necessary to further describe how to arrange RWB, oil palm seeds, and the media used in the container. Is RWb mixed with the media, or how? 4. Results, point 3.2. This subtitle is a bit inappropriate, because the content is a description of the morphology of the Ganoderma fruit body found. It would be better if the title were "Morphological characteristic......" 5. Results, Figure 2. In part (B), it is stated as the Initial stage of infection and the pinhead of the Ganoderma mushroom. This is a less appropriate statement, because the initial infection of Ganoderma on oil palm is not related to the appearance of pinheads. The appearance of pinheads is a sign that the infection is advanced and the disease is severe. 6. Figure 5. The image is not clear. The 0.5 mm line mark does not seem to match the original condition. In part (ii), root decay, necrosis, and decaying bole tissue are not visible. If possible, the image can be displayed at a closer up, so that the message to be conveyed can be better received by the reader. 7. Table 2. If possible, it would be better if the disease severity data could be displayed. 8. Figure 6. Not necessary. This image shows nothing. 9. It would be better if a general discussion were added that supports the main findings. For example, the condition of the environmental vegetation around the sample plantation is suspected of causing G. ryvardenii to be there and attack oil palms. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? Partly Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests: No competing interests were disclosed. Reviewer Expertise: Plantation crop cultivation, plant disease, oil palm disease, and biological control of plant disease. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Supriyanto S. Reviewer Report For: First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved, 2 approved with reservations] . F1000Research 2025, 14 :413 ( https://doi.org/10.5256/f1000research.178088.r377597 ) The direct URL for this report is: https://f1000research.com/articles/14-413/v1#referee-response-377597 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Author Response 11 Sep 2025 Emmanuellah Lekete-Lawson , Dep. of Microbiology, Czech University of Life Sciences Prague, Prague, Czech Republic 11 Sep 2025 Author Response Response to Reviewer’s comment Reviewer 1 Reviewer 1: Overall, this article is well written. However, some parts need to be strengthened to make it a reliable reference. Comment ... Continue reading Response to Reviewer’s comment Reviewer 1 Reviewer 1: Overall, this article is well written. However, some parts need to be strengthened to make it a reliable reference. Comment 1. In the last paragraph of the introduction, considering the author's claim about the discovery of a new species of Ganoderma that causes Basal Stem Rot disease in oil palm, it is better to add a new, more complete reference about reports related to the types of Ganoderma that previous researchers have reported. Response1: If I understand your comment, I believe you want us to elaborate on the discovery work conducted by previous researchers concerning the species identified in the current study. If so, here is the reference to the work done by: Researchers (Kinge TR and Mih AM), In 2010 and 2011 these researchers conducted a comprehensive study in the littoral and southwestern regions of Cameroon to identify Ganoderma species associated with basal stem rot (BSR) disease of oil palm. During the survey, these researchers encountered a previously unclassified Ganoderma species, which morphological and molecular analyses revealed to be distinct from all known species ever reported. To determine its identity, a morphological and molecular characterisation of the ITS rDNA locus was performed, which revealed variation in ellipsoidal basidiospores with narrowly truncate apices, thereby differentiating it from its close relatives like G. steyaertanum and the G. boninense clade, but it stands as a strongly supported standalone lineage. Expressing their admiration for its distinctiveness, the researchers named it Ganoderma ryvardense, in appreciation of the extraordinary mycologist Leif Ryvarden, whose outstanding work in African fungal taxonomy cannot be ignored. (This is inserted on page 3 of the revised manuscript.) Comment-2. Method . Observation of disease severity in oil palm in the field uses a score of 0 to 4 . The scoring criteria should be displayed in full . In addition , the use of the number of fruiting bodies as an indicator of severity seems rather risky because there has been no report on this. Based on experience in Indonesia, the number of fruiting bodies that can be found is not directly related to the severity of the disease. Response 2a: The Disease Scoring Criteria is displayed on page 5 of the manuscript as seen in the table below: Disease severity score Disease severity was scored using a scale of 0 to 4 (Lekete-Lawson et al. , (2024); Guide Disease severity index for Ganoderma infection (BSR) of oil palm Severity score No. of BSR fruiting bodies/foliar symptoms /plant Percent diseased palm 0 No BSR fruiting body/no foliar symptoms Healthy plant or no disease 1 1– 3 BSR fruiting body/foliar symptoms on two fronds 1– 25 % 2 4– 6 (BSR) fruiting body/foliar symptoms on three fronds 26– 50% 3 7– 9 (BSR) fruiting body/foliar symptoms on four fronds 51– 75% 4 > 9 (BSR) fruiting body/foliar symptoms on > four fronds 76 –100% Response 2b. Reasons of using fruiting bodies as an indicator of severity: The use of fruiting bodies as a severity indicator was adopted for the purpose of this study and was derived from the idea in the mushroom production technique. Ganoderma pathogen is a white rot fungal disease and difficult to diagnose at early stage and can also be misdiagnosed if using only foliar symptoms of the infected plant alone, less you see the mushroom which is the sign depicting the surest presence of the disease. Disease severity measures how badly the plant is affected, not just whether it is infected or how many of the plants are infected (incidence). The amount of mushrooms present per spot gives us an idea of how severely a plant is damaged as a result of the infection and the presence of the inoculum load. Also, disease severity focuses on the Intensity of the damage (e.g., % of area covered with lesions) or the percentage plant part (stem) covered with the mushroom; hence, the amount of mushroom coming out from the stem gives us an idea of how severely that plant is infected. How intense is the infection? This is the first time an intensive study has been conducted since the outbreak of Ganoderma Basal stem Rot disease in Ghana; we do not have sophisticated instruments/tools to detect how severely the plant is affected than to use the visual sign of the disease to show. Comment 3: Method : pathogenicity test. To be clearer, it is necessary to further describe how to arrange RWB, oil palm seeds, and the media used in the container. Is RWb mixed with the media, or how ? Response 3: In this experiment, we have stated that due to the scarcity of the Rubber Wood Block (RWB), we improvised RWB with Wawa wood block (WWB), not RWB. We adopted techniques by Angel et al. (2021) and Bivi et al., ( 2016 ), and modified to suit our study and in each treatment bottle, the mixture of the growing media in the bottle were also clearly stated under the methodology. (A volume of 250 ml Beatson glass jars (R3/83 mm 4 Doz) containing a mixture of planting media (black soil, bio-cha, and treated plant biostimulant) at the ratio of 2:2:1 100 g per glass was used. The jars were autoclaved for 45 minutes at 121°C. The mixtures were left to cool for three days) Comment 4 : Results , point 3.2. This subtitle is a bit inappropriate, because the content is a description of the morphology of the Ganoderma fruit body found. It would be better if the title were " Morphological characteristic......" Response 4: The title Physiological characteristics is replaced with morphological characteristics as suggested on page 8. Thanks. Comment 5. Results, Figure 2. In part (B), it is stated as the Initial stage of infection and the pinhead of the Ganoderma mushroom. This is a less appropriate statement, because the initial infection of Ganoderma on oil palm is not related to the appearance of pinheads. The appearance of pinheads is a sign that the infection is advanced and the disease is severe. Response 5: The pinhead is the first visible sign of the mushroom or the initial sign of the infection. Basal stem rot disease is often accompanied by the formation of the fruiting bodies on the lower/upper trunk and exposed root area as depicted in the picture (Fig. 2), In our case foliar symptoms were not prominent hence the appearance of the mushroom (pinhead was the only indication of the presence of the pathogen showing initial sign of the disease) This could probably be unique with this particular species “no foliar symptom but presence of the mushroom”. Comment 6. Figure 5. The image is not clear. The 0.5 mm line mark does not seem to match the original condition . In part (ii), root decay, necrosis, and decaying bole tissue are not visible. If possible, the image can be displayed closer up, so that the reader can better receive the message to be conveyed. Res: 6 The image has been replaced on page 11. However, the experiment was done on the two-week-old germinated seed nuts; hence, the bole of the seed nut was not visibly pronounced as compared to if the experiment were to be done on one or two-month-old seedlings. Comment 7 . Table 2. If possible, it would be better if the disease severity data could be displayed. Response 7: The disease severity data was provided in the raw dataset repository at (figshare.com repository: Lekete-Lawson E: Report on Basal Stem Rot (BSR) disease on oil palm in Ghana. figshare. Journal contribution. 2025b. 10.6084/m9.figshare.28329830.v1); for further explanation and understanding, however, to avoid overcrowding we chose to present the ANOVA table in the results section. Comment 8 . Figure 6. Not necessary. This image shows nothing. Response 8: Figure 6 is removed (page 12) Comment 9. It would be better if a general discussion were added that supports the main findings. For example, the environmental vegetation around the sample plantation is suspected of causing G. ryvardenii to be there and attack oil palms. Response 9: This is the initial study and the first report; the study is ongoing, although there have never been any report of G. ryvadenii associated with the type vegetation of vegetation we have or any tree crops or oil palm in Ghana in the past, however subsequent studies into other factors including the environmental and host factors that could lead to the presence of G. ryvadenii is ongoing. Meanwhile, the climatic conditions of the individual study areas are well stated under the climate of the study areas in the methodology Page 4. Response to Reviewer’s comment Reviewer 1 Reviewer 1: Overall, this article is well written. However, some parts need to be strengthened to make it a reliable reference. Comment 1. In the last paragraph of the introduction, considering the author's claim about the discovery of a new species of Ganoderma that causes Basal Stem Rot disease in oil palm, it is better to add a new, more complete reference about reports related to the types of Ganoderma that previous researchers have reported. Response1: If I understand your comment, I believe you want us to elaborate on the discovery work conducted by previous researchers concerning the species identified in the current study. If so, here is the reference to the work done by: Researchers (Kinge TR and Mih AM), In 2010 and 2011 these researchers conducted a comprehensive study in the littoral and southwestern regions of Cameroon to identify Ganoderma species associated with basal stem rot (BSR) disease of oil palm. During the survey, these researchers encountered a previously unclassified Ganoderma species, which morphological and molecular analyses revealed to be distinct from all known species ever reported. To determine its identity, a morphological and molecular characterisation of the ITS rDNA locus was performed, which revealed variation in ellipsoidal basidiospores with narrowly truncate apices, thereby differentiating it from its close relatives like G. steyaertanum and the G. boninense clade, but it stands as a strongly supported standalone lineage. Expressing their admiration for its distinctiveness, the researchers named it Ganoderma ryvardense, in appreciation of the extraordinary mycologist Leif Ryvarden, whose outstanding work in African fungal taxonomy cannot be ignored. (This is inserted on page 3 of the revised manuscript.) Comment-2. Method . Observation of disease severity in oil palm in the field uses a score of 0 to 4 . The scoring criteria should be displayed in full . In addition , the use of the number of fruiting bodies as an indicator of severity seems rather risky because there has been no report on this. Based on experience in Indonesia, the number of fruiting bodies that can be found is not directly related to the severity of the disease. Response 2a: The Disease Scoring Criteria is displayed on page 5 of the manuscript as seen in the table below: Disease severity score Disease severity was scored using a scale of 0 to 4 (Lekete-Lawson et al. , (2024); Guide Disease severity index for Ganoderma infection (BSR) of oil palm Severity score No. of BSR fruiting bodies/foliar symptoms /plant Percent diseased palm 0 No BSR fruiting body/no foliar symptoms Healthy plant or no disease 1 1– 3 BSR fruiting body/foliar symptoms on two fronds 1– 25 % 2 4– 6 (BSR) fruiting body/foliar symptoms on three fronds 26– 50% 3 7– 9 (BSR) fruiting body/foliar symptoms on four fronds 51– 75% 4 > 9 (BSR) fruiting body/foliar symptoms on > four fronds 76 –100% Response 2b. Reasons of using fruiting bodies as an indicator of severity: The use of fruiting bodies as a severity indicator was adopted for the purpose of this study and was derived from the idea in the mushroom production technique. Ganoderma pathogen is a white rot fungal disease and difficult to diagnose at early stage and can also be misdiagnosed if using only foliar symptoms of the infected plant alone, less you see the mushroom which is the sign depicting the surest presence of the disease. Disease severity measures how badly the plant is affected, not just whether it is infected or how many of the plants are infected (incidence). The amount of mushrooms present per spot gives us an idea of how severely a plant is damaged as a result of the infection and the presence of the inoculum load. Also, disease severity focuses on the Intensity of the damage (e.g., % of area covered with lesions) or the percentage plant part (stem) covered with the mushroom; hence, the amount of mushroom coming out from the stem gives us an idea of how severely that plant is infected. How intense is the infection? This is the first time an intensive study has been conducted since the outbreak of Ganoderma Basal stem Rot disease in Ghana; we do not have sophisticated instruments/tools to detect how severely the plant is affected than to use the visual sign of the disease to show. Comment 3: Method : pathogenicity test. To be clearer, it is necessary to further describe how to arrange RWB, oil palm seeds, and the media used in the container. Is RWb mixed with the media, or how ? Response 3: In this experiment, we have stated that due to the scarcity of the Rubber Wood Block (RWB), we improvised RWB with Wawa wood block (WWB), not RWB. We adopted techniques by Angel et al. (2021) and Bivi et al., ( 2016 ), and modified to suit our study and in each treatment bottle, the mixture of the growing media in the bottle were also clearly stated under the methodology. (A volume of 250 ml Beatson glass jars (R3/83 mm 4 Doz) containing a mixture of planting media (black soil, bio-cha, and treated plant biostimulant) at the ratio of 2:2:1 100 g per glass was used. The jars were autoclaved for 45 minutes at 121°C. The mixtures were left to cool for three days) Comment 4 : Results , point 3.2. This subtitle is a bit inappropriate, because the content is a description of the morphology of the Ganoderma fruit body found. It would be better if the title were " Morphological characteristic......" Response 4: The title Physiological characteristics is replaced with morphological characteristics as suggested on page 8. Thanks. Comment 5. Results, Figure 2. In part (B), it is stated as the Initial stage of infection and the pinhead of the Ganoderma mushroom. This is a less appropriate statement, because the initial infection of Ganoderma on oil palm is not related to the appearance of pinheads. The appearance of pinheads is a sign that the infection is advanced and the disease is severe. Response 5: The pinhead is the first visible sign of the mushroom or the initial sign of the infection. Basal stem rot disease is often accompanied by the formation of the fruiting bodies on the lower/upper trunk and exposed root area as depicted in the picture (Fig. 2), In our case foliar symptoms were not prominent hence the appearance of the mushroom (pinhead was the only indication of the presence of the pathogen showing initial sign of the disease) This could probably be unique with this particular species “no foliar symptom but presence of the mushroom”. Comment 6. Figure 5. The image is not clear. The 0.5 mm line mark does not seem to match the original condition . In part (ii), root decay, necrosis, and decaying bole tissue are not visible. If possible, the image can be displayed closer up, so that the reader can better receive the message to be conveyed. Res: 6 The image has been replaced on page 11. However, the experiment was done on the two-week-old germinated seed nuts; hence, the bole of the seed nut was not visibly pronounced as compared to if the experiment were to be done on one or two-month-old seedlings. Comment 7 . Table 2. If possible, it would be better if the disease severity data could be displayed. Response 7: The disease severity data was provided in the raw dataset repository at (figshare.com repository: Lekete-Lawson E: Report on Basal Stem Rot (BSR) disease on oil palm in Ghana. figshare. Journal contribution. 2025b. 10.6084/m9.figshare.28329830.v1); for further explanation and understanding, however, to avoid overcrowding we chose to present the ANOVA table in the results section. Comment 8 . Figure 6. Not necessary. This image shows nothing. Response 8: Figure 6 is removed (page 12) Comment 9. It would be better if a general discussion were added that supports the main findings. For example, the environmental vegetation around the sample plantation is suspected of causing G. ryvardenii to be there and attack oil palms. Response 9: This is the initial study and the first report; the study is ongoing, although there have never been any report of G. ryvadenii associated with the type vegetation of vegetation we have or any tree crops or oil palm in Ghana in the past, however subsequent studies into other factors including the environmental and host factors that could lead to the presence of G. ryvadenii is ongoing. Meanwhile, the climatic conditions of the individual study areas are well stated under the climate of the study areas in the methodology Page 4. Competing Interests: Authors declare no competing interest Close Report a concern Respond or Comment COMMENTS ON THIS REPORT Author Response 11 Sep 2025 Emmanuellah Lekete-Lawson , Dep. of Microbiology, Czech University of Life Sciences Prague, Prague, Czech Republic 11 Sep 2025 Author Response Response to Reviewer’s comment Reviewer 1 Reviewer 1: Overall, this article is well written. However, some parts need to be strengthened to make it a reliable reference. Comment ... Continue reading Response to Reviewer’s comment Reviewer 1 Reviewer 1: Overall, this article is well written. However, some parts need to be strengthened to make it a reliable reference. Comment 1. In the last paragraph of the introduction, considering the author's claim about the discovery of a new species of Ganoderma that causes Basal Stem Rot disease in oil palm, it is better to add a new, more complete reference about reports related to the types of Ganoderma that previous researchers have reported. Response1: If I understand your comment, I believe you want us to elaborate on the discovery work conducted by previous researchers concerning the species identified in the current study. If so, here is the reference to the work done by: Researchers (Kinge TR and Mih AM), In 2010 and 2011 these researchers conducted a comprehensive study in the littoral and southwestern regions of Cameroon to identify Ganoderma species associated with basal stem rot (BSR) disease of oil palm. During the survey, these researchers encountered a previously unclassified Ganoderma species, which morphological and molecular analyses revealed to be distinct from all known species ever reported. To determine its identity, a morphological and molecular characterisation of the ITS rDNA locus was performed, which revealed variation in ellipsoidal basidiospores with narrowly truncate apices, thereby differentiating it from its close relatives like G. steyaertanum and the G. boninense clade, but it stands as a strongly supported standalone lineage. Expressing their admiration for its distinctiveness, the researchers named it Ganoderma ryvardense, in appreciation of the extraordinary mycologist Leif Ryvarden, whose outstanding work in African fungal taxonomy cannot be ignored. (This is inserted on page 3 of the revised manuscript.) Comment-2. Method . Observation of disease severity in oil palm in the field uses a score of 0 to 4 . The scoring criteria should be displayed in full . In addition , the use of the number of fruiting bodies as an indicator of severity seems rather risky because there has been no report on this. Based on experience in Indonesia, the number of fruiting bodies that can be found is not directly related to the severity of the disease. Response 2a: The Disease Scoring Criteria is displayed on page 5 of the manuscript as seen in the table below: Disease severity score Disease severity was scored using a scale of 0 to 4 (Lekete-Lawson et al. , (2024); Guide Disease severity index for Ganoderma infection (BSR) of oil palm Severity score No. of BSR fruiting bodies/foliar symptoms /plant Percent diseased palm 0 No BSR fruiting body/no foliar symptoms Healthy plant or no disease 1 1– 3 BSR fruiting body/foliar symptoms on two fronds 1– 25 % 2 4– 6 (BSR) fruiting body/foliar symptoms on three fronds 26– 50% 3 7– 9 (BSR) fruiting body/foliar symptoms on four fronds 51– 75% 4 > 9 (BSR) fruiting body/foliar symptoms on > four fronds 76 –100% Response 2b. Reasons of using fruiting bodies as an indicator of severity: The use of fruiting bodies as a severity indicator was adopted for the purpose of this study and was derived from the idea in the mushroom production technique. Ganoderma pathogen is a white rot fungal disease and difficult to diagnose at early stage and can also be misdiagnosed if using only foliar symptoms of the infected plant alone, less you see the mushroom which is the sign depicting the surest presence of the disease. Disease severity measures how badly the plant is affected, not just whether it is infected or how many of the plants are infected (incidence). The amount of mushrooms present per spot gives us an idea of how severely a plant is damaged as a result of the infection and the presence of the inoculum load. Also, disease severity focuses on the Intensity of the damage (e.g., % of area covered with lesions) or the percentage plant part (stem) covered with the mushroom; hence, the amount of mushroom coming out from the stem gives us an idea of how severely that plant is infected. How intense is the infection? This is the first time an intensive study has been conducted since the outbreak of Ganoderma Basal stem Rot disease in Ghana; we do not have sophisticated instruments/tools to detect how severely the plant is affected than to use the visual sign of the disease to show. Comment 3: Method : pathogenicity test. To be clearer, it is necessary to further describe how to arrange RWB, oil palm seeds, and the media used in the container. Is RWb mixed with the media, or how ? Response 3: In this experiment, we have stated that due to the scarcity of the Rubber Wood Block (RWB), we improvised RWB with Wawa wood block (WWB), not RWB. We adopted techniques by Angel et al. (2021) and Bivi et al., ( 2016 ), and modified to suit our study and in each treatment bottle, the mixture of the growing media in the bottle were also clearly stated under the methodology. (A volume of 250 ml Beatson glass jars (R3/83 mm 4 Doz) containing a mixture of planting media (black soil, bio-cha, and treated plant biostimulant) at the ratio of 2:2:1 100 g per glass was used. The jars were autoclaved for 45 minutes at 121°C. The mixtures were left to cool for three days) Comment 4 : Results , point 3.2. This subtitle is a bit inappropriate, because the content is a description of the morphology of the Ganoderma fruit body found. It would be better if the title were " Morphological characteristic......" Response 4: The title Physiological characteristics is replaced with morphological characteristics as suggested on page 8. Thanks. Comment 5. Results, Figure 2. In part (B), it is stated as the Initial stage of infection and the pinhead of the Ganoderma mushroom. This is a less appropriate statement, because the initial infection of Ganoderma on oil palm is not related to the appearance of pinheads. The appearance of pinheads is a sign that the infection is advanced and the disease is severe. Response 5: The pinhead is the first visible sign of the mushroom or the initial sign of the infection. Basal stem rot disease is often accompanied by the formation of the fruiting bodies on the lower/upper trunk and exposed root area as depicted in the picture (Fig. 2), In our case foliar symptoms were not prominent hence the appearance of the mushroom (pinhead was the only indication of the presence of the pathogen showing initial sign of the disease) This could probably be unique with this particular species “no foliar symptom but presence of the mushroom”. Comment 6. Figure 5. The image is not clear. The 0.5 mm line mark does not seem to match the original condition . In part (ii), root decay, necrosis, and decaying bole tissue are not visible. If possible, the image can be displayed closer up, so that the reader can better receive the message to be conveyed. Res: 6 The image has been replaced on page 11. However, the experiment was done on the two-week-old germinated seed nuts; hence, the bole of the seed nut was not visibly pronounced as compared to if the experiment were to be done on one or two-month-old seedlings. Comment 7 . Table 2. If possible, it would be better if the disease severity data could be displayed. Response 7: The disease severity data was provided in the raw dataset repository at (figshare.com repository: Lekete-Lawson E: Report on Basal Stem Rot (BSR) disease on oil palm in Ghana. figshare. Journal contribution. 2025b. 10.6084/m9.figshare.28329830.v1); for further explanation and understanding, however, to avoid overcrowding we chose to present the ANOVA table in the results section. Comment 8 . Figure 6. Not necessary. This image shows nothing. Response 8: Figure 6 is removed (page 12) Comment 9. It would be better if a general discussion were added that supports the main findings. For example, the environmental vegetation around the sample plantation is suspected of causing G. ryvardenii to be there and attack oil palms. Response 9: This is the initial study and the first report; the study is ongoing, although there have never been any report of G. ryvadenii associated with the type vegetation of vegetation we have or any tree crops or oil palm in Ghana in the past, however subsequent studies into other factors including the environmental and host factors that could lead to the presence of G. ryvadenii is ongoing. Meanwhile, the climatic conditions of the individual study areas are well stated under the climate of the study areas in the methodology Page 4. Response to Reviewer’s comment Reviewer 1 Reviewer 1: Overall, this article is well written. However, some parts need to be strengthened to make it a reliable reference. Comment 1. In the last paragraph of the introduction, considering the author's claim about the discovery of a new species of Ganoderma that causes Basal Stem Rot disease in oil palm, it is better to add a new, more complete reference about reports related to the types of Ganoderma that previous researchers have reported. Response1: If I understand your comment, I believe you want us to elaborate on the discovery work conducted by previous researchers concerning the species identified in the current study. If so, here is the reference to the work done by: Researchers (Kinge TR and Mih AM), In 2010 and 2011 these researchers conducted a comprehensive study in the littoral and southwestern regions of Cameroon to identify Ganoderma species associated with basal stem rot (BSR) disease of oil palm. During the survey, these researchers encountered a previously unclassified Ganoderma species, which morphological and molecular analyses revealed to be distinct from all known species ever reported. To determine its identity, a morphological and molecular characterisation of the ITS rDNA locus was performed, which revealed variation in ellipsoidal basidiospores with narrowly truncate apices, thereby differentiating it from its close relatives like G. steyaertanum and the G. boninense clade, but it stands as a strongly supported standalone lineage. Expressing their admiration for its distinctiveness, the researchers named it Ganoderma ryvardense, in appreciation of the extraordinary mycologist Leif Ryvarden, whose outstanding work in African fungal taxonomy cannot be ignored. (This is inserted on page 3 of the revised manuscript.) Comment-2. Method . Observation of disease severity in oil palm in the field uses a score of 0 to 4 . The scoring criteria should be displayed in full . In addition , the use of the number of fruiting bodies as an indicator of severity seems rather risky because there has been no report on this. Based on experience in Indonesia, the number of fruiting bodies that can be found is not directly related to the severity of the disease. Response 2a: The Disease Scoring Criteria is displayed on page 5 of the manuscript as seen in the table below: Disease severity score Disease severity was scored using a scale of 0 to 4 (Lekete-Lawson et al. , (2024); Guide Disease severity index for Ganoderma infection (BSR) of oil palm Severity score No. of BSR fruiting bodies/foliar symptoms /plant Percent diseased palm 0 No BSR fruiting body/no foliar symptoms Healthy plant or no disease 1 1– 3 BSR fruiting body/foliar symptoms on two fronds 1– 25 % 2 4– 6 (BSR) fruiting body/foliar symptoms on three fronds 26– 50% 3 7– 9 (BSR) fruiting body/foliar symptoms on four fronds 51– 75% 4 > 9 (BSR) fruiting body/foliar symptoms on > four fronds 76 –100% Response 2b. Reasons of using fruiting bodies as an indicator of severity: The use of fruiting bodies as a severity indicator was adopted for the purpose of this study and was derived from the idea in the mushroom production technique. Ganoderma pathogen is a white rot fungal disease and difficult to diagnose at early stage and can also be misdiagnosed if using only foliar symptoms of the infected plant alone, less you see the mushroom which is the sign depicting the surest presence of the disease. Disease severity measures how badly the plant is affected, not just whether it is infected or how many of the plants are infected (incidence). The amount of mushrooms present per spot gives us an idea of how severely a plant is damaged as a result of the infection and the presence of the inoculum load. Also, disease severity focuses on the Intensity of the damage (e.g., % of area covered with lesions) or the percentage plant part (stem) covered with the mushroom; hence, the amount of mushroom coming out from the stem gives us an idea of how severely that plant is infected. How intense is the infection? This is the first time an intensive study has been conducted since the outbreak of Ganoderma Basal stem Rot disease in Ghana; we do not have sophisticated instruments/tools to detect how severely the plant is affected than to use the visual sign of the disease to show. Comment 3: Method : pathogenicity test. To be clearer, it is necessary to further describe how to arrange RWB, oil palm seeds, and the media used in the container. Is RWb mixed with the media, or how ? Response 3: In this experiment, we have stated that due to the scarcity of the Rubber Wood Block (RWB), we improvised RWB with Wawa wood block (WWB), not RWB. We adopted techniques by Angel et al. (2021) and Bivi et al., ( 2016 ), and modified to suit our study and in each treatment bottle, the mixture of the growing media in the bottle were also clearly stated under the methodology. (A volume of 250 ml Beatson glass jars (R3/83 mm 4 Doz) containing a mixture of planting media (black soil, bio-cha, and treated plant biostimulant) at the ratio of 2:2:1 100 g per glass was used. The jars were autoclaved for 45 minutes at 121°C. The mixtures were left to cool for three days) Comment 4 : Results , point 3.2. This subtitle is a bit inappropriate, because the content is a description of the morphology of the Ganoderma fruit body found. It would be better if the title were " Morphological characteristic......" Response 4: The title Physiological characteristics is replaced with morphological characteristics as suggested on page 8. Thanks. Comment 5. Results, Figure 2. In part (B), it is stated as the Initial stage of infection and the pinhead of the Ganoderma mushroom. This is a less appropriate statement, because the initial infection of Ganoderma on oil palm is not related to the appearance of pinheads. The appearance of pinheads is a sign that the infection is advanced and the disease is severe. Response 5: The pinhead is the first visible sign of the mushroom or the initial sign of the infection. Basal stem rot disease is often accompanied by the formation of the fruiting bodies on the lower/upper trunk and exposed root area as depicted in the picture (Fig. 2), In our case foliar symptoms were not prominent hence the appearance of the mushroom (pinhead was the only indication of the presence of the pathogen showing initial sign of the disease) This could probably be unique with this particular species “no foliar symptom but presence of the mushroom”. Comment 6. Figure 5. The image is not clear. The 0.5 mm line mark does not seem to match the original condition . In part (ii), root decay, necrosis, and decaying bole tissue are not visible. If possible, the image can be displayed closer up, so that the reader can better receive the message to be conveyed. Res: 6 The image has been replaced on page 11. However, the experiment was done on the two-week-old germinated seed nuts; hence, the bole of the seed nut was not visibly pronounced as compared to if the experiment were to be done on one or two-month-old seedlings. Comment 7 . Table 2. If possible, it would be better if the disease severity data could be displayed. Response 7: The disease severity data was provided in the raw dataset repository at (figshare.com repository: Lekete-Lawson E: Report on Basal Stem Rot (BSR) disease on oil palm in Ghana. figshare. Journal contribution. 2025b. 10.6084/m9.figshare.28329830.v1); for further explanation and understanding, however, to avoid overcrowding we chose to present the ANOVA table in the results section. Comment 8 . Figure 6. Not necessary. This image shows nothing. Response 8: Figure 6 is removed (page 12) Comment 9. It would be better if a general discussion were added that supports the main findings. For example, the environmental vegetation around the sample plantation is suspected of causing G. ryvardenii to be there and attack oil palms. Response 9: This is the initial study and the first report; the study is ongoing, although there have never been any report of G. ryvadenii associated with the type vegetation of vegetation we have or any tree crops or oil palm in Ghana in the past, however subsequent studies into other factors including the environmental and host factors that could lead to the presence of G. ryvadenii is ongoing. Meanwhile, the climatic conditions of the individual study areas are well stated under the climate of the study areas in the methodology Page 4. Competing Interests: Authors declare no competing interest Close Report a concern COMMENT ON THIS REPORT Comments on this article Comments (1) Version 2 VERSION 2 PUBLISHED 07 Jul 2025 Revised Comment ADD YOUR COMMENT Version 1 VERSION 1 PUBLISHED 08 Apr 2025 Discussion is closed on this version, please comment on the latest version above. Reader Comment 04 May 2025 Robert Paterson , Centre for Biological Engineering, University of Minho, Braga, Portugal 04 May 2025 Reader Comment Comment on Lekete-Lawson et al. (2025) By: Robert Russell Monteith Paterson University of Minho, Portugal [email protected] I was interested to read the pre-review paper on Ganoderma from oil ... Continue reading Comment on Lekete-Lawson et al. (2025) By: Robert Russell Monteith Paterson University of Minho, Portugal [email protected] I was interested to read the pre-review paper on Ganoderma from oil palm in Ghana (Lekete-Lawson et al. 2025) having published quiet extensively in the field in other countries. It is worth reviewing the current situation. G. boninense is considered the basal stem rot (BSR) fungus in Southeast (SE) Asia and Oceania where Indonesia and Malaysia are by far the largest producers of palm oil. G. zonatum is also considered to cause BSR and a paper recently claimed that G. zonatum is the cause of BSR in Colombia, South America. G.ryvardenii is related closely to, but distinct from, G boninense and G. zonatum . Clearly, G. zonatum and G. boninense are also closely related. My first concern when reading Lekete-Lawson et al. (2025) was the PCR analysis and subsequently I detected other problems. PCR In relation to the PCR, in the text it says Figure 7A is for the culture based samples but these are Figure 7B in the legend to the image of the gel. Figure 7 B is the text is for basidiocarps but these are 7A in the legend to the figure. Which is it? Also, there is no information about the strain numbers that represented tracks 1 to 24 in the figure of the gels. These should be provided. It is all very confused. Also, there appeared to be a faint band in the blanks for samples 1 to 24 which were at the same intensity for track 20. Presumably the blanks were contaminated. Were tracks 20 and 35 from the same specimen and strain? Was there a procedure for identifying inhibition of the PCR? I know that an internal amplification control (IAC) to assess inhibition is recommended for at least some PCRs. IAC would provide information on inhibition or could it be that the samples represented by 20 and 35 were not what you thought they were? Why did you not run known relevant Ganoderma species for comparison with the other specimens? This would provide information on the novelty of your specimens. Overall, there are considerable problems with the presentation of the PCR results. Other concerns are as follows: General A strain and specimen list is necessary with accession numbers. Are you saying G. ryvardenii is the only species involved in BSR in Ghana? What is the new species you say you have discovered? This is unclear. A considerable amount of this paper is taken from a previous paper (Lekete-Lawson et al. 2024). This paper should be discussed in the introduction and cited properly. And any duplication needs to be removed. Somebody who is excellent at writing scientific papers in English should be asked to rewrite the paper. Abstract Palm oil is not necessarily the foremost vegetable oil. For example, soy oil is arguably more important. The abstract is vague. What do you mean by using nucleic acid as a template? The English is poor, e.g. “BSR is characterised by stem decay large-perennial, woody brackets basidiocarps”. What does this mean? The conclusion does not make sense to me at all. Introduction India is only a small palm oil producing country in contradiction to what you have written. You say Africa produces 3% oil palm and then West Africa produces 4%. Which is it? G. boninense is the most serious disease of oil palm in SE Asia and Oceania. In Africa it is considered that fusarium wilt is the most serious fungal disease. G. boninense is accepted as the fungus responsible for BSR in SE Asia at least. You need to mention that G. ryvardenii was identified in Cameroon before and give the citation. Material and Methods Figure 1 should be in the results. In the sampling and isolation section, what does “including fruiting bodies (Basidiocarps) and Ganoderma sp. (Dikaryotic)” mean? How were specimens tested for purity? What does “Microzone, UK, proprietary formulation microLYSIS®-PLUS (MLP) underwent rapid thermal cycling for cell lysis and release of DNA” mean? GanET is G. boninense specific (or better, a palm clade specific) primer. It is not basidiomycete specific. Please correct this. Results Did you test the difference between upper stem rot specimens and basal stem rot specimens? These have been considered as different species before. Phytophthora is not considered to be a fungus. It is an oomycete. The Phylogenetic results section is poor with inadequate information. Discussion Cameroon is not in west Africa. References Lekete-Lawson E, van der Puije GC, Osekre EA and Ackah FK. First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved with reservations]. F1000Research 2025, 14 :413 ( https://doi.org/10.12688/f1000research.161972.1 ) Lekete-Lawson, E, Osekre, E.A. van der Puije, G., Swanzy, et al. 2024. Prevalence of basal stem rot disease caused by Ganoderma sp. on oil palm ( Elaeis guineensis Jacq) in Ghana. Journal of Science and Technology, Special Issue No 1, pp 47 – 61. Comment on Lekete-Lawson et al. (2025) By: Robert Russell Monteith Paterson University of Minho, Portugal [email protected] I was interested to read the pre-review paper on Ganoderma from oil palm in Ghana (Lekete-Lawson et al. 2025) having published quiet extensively in the field in other countries. It is worth reviewing the current situation. G. boninense is considered the basal stem rot (BSR) fungus in Southeast (SE) Asia and Oceania where Indonesia and Malaysia are by far the largest producers of palm oil. G. zonatum is also considered to cause BSR and a paper recently claimed that G. zonatum is the cause of BSR in Colombia, South America. G.ryvardenii is related closely to, but distinct from, G boninense and G. zonatum . Clearly, G. zonatum and G. boninense are also closely related. My first concern when reading Lekete-Lawson et al. (2025) was the PCR analysis and subsequently I detected other problems. PCR In relation to the PCR, in the text it says Figure 7A is for the culture based samples but these are Figure 7B in the legend to the image of the gel. Figure 7 B is the text is for basidiocarps but these are 7A in the legend to the figure. Which is it? Also, there is no information about the strain numbers that represented tracks 1 to 24 in the figure of the gels. These should be provided. It is all very confused. Also, there appeared to be a faint band in the blanks for samples 1 to 24 which were at the same intensity for track 20. Presumably the blanks were contaminated. Were tracks 20 and 35 from the same specimen and strain? Was there a procedure for identifying inhibition of the PCR? I know that an internal amplification control (IAC) to assess inhibition is recommended for at least some PCRs. IAC would provide information on inhibition or could it be that the samples represented by 20 and 35 were not what you thought they were? Why did you not run known relevant Ganoderma species for comparison with the other specimens? This would provide information on the novelty of your specimens. Overall, there are considerable problems with the presentation of the PCR results. Other concerns are as follows: General A strain and specimen list is necessary with accession numbers. Are you saying G. ryvardenii is the only species involved in BSR in Ghana? What is the new species you say you have discovered? This is unclear. A considerable amount of this paper is taken from a previous paper (Lekete-Lawson et al. 2024). This paper should be discussed in the introduction and cited properly. And any duplication needs to be removed. Somebody who is excellent at writing scientific papers in English should be asked to rewrite the paper. Abstract Palm oil is not necessarily the foremost vegetable oil. For example, soy oil is arguably more important. The abstract is vague. What do you mean by using nucleic acid as a template? The English is poor, e.g. “BSR is characterised by stem decay large-perennial, woody brackets basidiocarps”. What does this mean? The conclusion does not make sense to me at all. Introduction India is only a small palm oil producing country in contradiction to what you have written. You say Africa produces 3% oil palm and then West Africa produces 4%. Which is it? G. boninense is the most serious disease of oil palm in SE Asia and Oceania. In Africa it is considered that fusarium wilt is the most serious fungal disease. G. boninense is accepted as the fungus responsible for BSR in SE Asia at least. You need to mention that G. ryvardenii was identified in Cameroon before and give the citation. Material and Methods Figure 1 should be in the results. In the sampling and isolation section, what does “including fruiting bodies (Basidiocarps) and Ganoderma sp. (Dikaryotic)” mean? How were specimens tested for purity? What does “Microzone, UK, proprietary formulation microLYSIS®-PLUS (MLP) underwent rapid thermal cycling for cell lysis and release of DNA” mean? GanET is G. boninense specific (or better, a palm clade specific) primer. It is not basidiomycete specific. Please correct this. Results Did you test the difference between upper stem rot specimens and basal stem rot specimens? These have been considered as different species before. Phytophthora is not considered to be a fungus. It is an oomycete. The Phylogenetic results section is poor with inadequate information. Discussion Cameroon is not in west Africa. References Lekete-Lawson E, van der Puije GC, Osekre EA and Ackah FK. First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved with reservations]. F1000Research 2025, 14 :413 ( https://doi.org/10.12688/f1000research.161972.1 ) Lekete-Lawson, E, Osekre, E.A. van der Puije, G., Swanzy, et al. 2024. Prevalence of basal stem rot disease caused by Ganoderma sp. on oil palm ( Elaeis guineensis Jacq) in Ghana. Journal of Science and Technology, Special Issue No 1, pp 47 – 61. Competing Interests: None Close Report a concern Discussion is closed on this version, please comment on the latest version above. keyboard_arrow_left keyboard_arrow_right Open Peer Review Reviewer Status info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Reviewer Reports Invited Reviewers 1 2 3 Version 2 (revision) 07 Jul 25 read Version 1 08 Apr 25 read read read Supriyanto Supriyanto , Tanjungpura University, Jl. Prof. Dr. H. Hadari Nawawi, Pontianak, Indonesia Efi Toding Tondok , IPB University, Bogor, Indonesia Lisnawita Lisnawita , Universitas Sumatera Utara,, Padang Bulan, Indonesia Comments on this article All Comments (1) Add a comment Sign up for content alerts Sign Up You are now signed up to receive this alert Browse by related subjects keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2025 Supriyanto S. This is an open access peer review report 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. 25 Jul 2025 | for Version 2 Supriyanto Supriyanto , Tanjungpura University, Jl. Prof. Dr. H. Hadari Nawawi, Pontianak, Indonesia 0 Views copyright © 2025 Supriyanto S. This is an open access peer review report 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. format_quote Cite this report speaker_notes Responses (0) Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions The manuscript has been well revised, but there are still several sections that have not been adjusted. These sections are: 1. Method (3.2). The authors have been changed to "Morphological characteristics....", but the results and discussion section (4.2) still uses the phrase "Physiological characteristics...."; this should be adjusted. 2. In the results and discussion. In Figure 2. "(B) Initial stage of infection and pinhead of the Ganoderma mushroom" is an inappropriate statement. It would be better to delete the phrase "Initial stage of infection" and use just the phrase "pinhead of the Ganoderma mushroom" because in this article, there is no evidence that the pinhead is the initial infection of Ganoderma. Researchers all state that the initial infection is more often through root contact. Furthermore, it would be better not to use the term "Ganoderma mushroom"; it would be more appropriate to use the term Ganoderma fruiting body or carpophore, which researchers commonly use. Competing Interests No competing interests were disclosed. Reviewer Expertise Plantation crop cultivation, plant disease, oil palm disease, and biological control of plant disease. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. reply Respond to this report Responses (0) Supriyanto S. Peer Review Report For: First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved, 2 approved with reservations] . F1000Research 2025, 14 :413 ( https://doi.org/10.5256/f1000research.184059.r397250) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/14-413/v2#referee-response-397250 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2025 Lisnawita L. This is an open access peer review report 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. 30 May 2025 | for Version 1 Lisnawita Lisnawita , Universitas Sumatera Utara,, Padang Bulan, Indonesia 0 Views copyright © 2025 Lisnawita L. This is an open access peer review report 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. format_quote Cite this report speaker_notes Responses (1) Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions In the method, it is not written how many oil palm plants were observed in the field research, but in the result is written 3000 oil palm trees were assessed. Figure 2, 4, 5, 6 are not clear Microscopic observations should be accompanied by Ganoderma spore images Researchers only used ITS and GanET for molecular identification. It is recommended to add additional genetic markers such as TEF1α or RPB2 for deeper validation, because the genus Ganoderma is known to have taxonomic complexity Conclusions should be stated explicitly. Do not state that the isolate is “probably” a new species. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise Phytopathology, Nematology I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. reply Respond to this report Responses (1) Author Response 11 Sep 2025 Emmanuellah Lekete-Lawson, Dep. of Microbiology, Czech University of Life Sciences Prague, Prague, Czech Republic Reviewer 3: Comment1: In the method, it is not written how many oil palm plants were observed in the field research, but in the result is written 3000 oil palm trees were assessed. Response1: It was stated in the Materials and methods that 30 oil palm communities/plantation were assessed and 100 palms per plantation, hence the total palms assessed were 3000 palms Comment 2: Figure 2, 4, 5, 6 are not clear Response 2 :6 was deleted the rest have been worked on Comment 3: Microscopic observations should be accompanied by Ganoderma spore images Response 3: Conscious effort was made to get the spore to aid the identification, but proof futile because of constant contamination and other minor Ganoderma species however the project is ongoing and plan to raise the mushroom invitro to reduce contamination is underway. Comment 4: Researchers only used ITS and GanET for molecular identification. It is recommended to add additional genetic markers such as TEF1α or RPB2 for deeper validation, because the genus Ganoderma is known to have taxonomic complexity Response 4: ITS and GanET were primers used by previous researchers, and this is our first time of working on Ganoderma, so we tried to follow the existing protocols/steps of the previous researchers for easy comparison and validation of our results. Comment 5: Conclusions should be stated explicitly. Do not state that the isolate is “probably” a new species. Response 5: Have been corrected. View more View less Competing Interests Authors declare no competing interest reply Respond Report a concern Lisnawita L. Peer Review Report For: First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved, 2 approved with reservations] . F1000Research 2025, 14 :413 ( https://doi.org/10.5256/f1000research.178088.r380554) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/14-413/v1#referee-response-380554 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2025 Tondok E. This is an open access peer review report 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. 19 May 2025 | for Version 1 Efi Toding Tondok , Plant Protection, IPB University, Bogor, West Java, Indonesia 0 Views copyright © 2025 Tondok E. This is an open access peer review report 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. format_quote Cite this report speaker_notes Responses (1) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Method . 1)Scores on disease severity must be written clearly. 2) This is Basidiomycota, so you will never find spores by mounting mycelia on object glass. Result . 1) Please add basidiospores morphometrics as one of the important features for morphological identification. The 0.5 mm bar is too long for microscopic measurement. 2) Fig 5 and 6: please put the objects in a good arrangement, so the reader will be easy to understand. Don't forget the control. The 0.5 mm bar is very annoying. 3) It is very important to include pictures from the fields, the mild or early symptom to severe or advance symptom. Formation of basidiocarp indicating the advance symptom, plant almost die. 4) How many species of Ganoderma found in Ghana? 5) Please put the same codes on your isolates since morphological identification - pathogenicity tests - molecular identification: easy to track it, to prove that the tested isolate is the same as the isolate identified morphologically and molecularly Discussion. Please explain why disease incidence and disease severity are highest in K29-B, compared to other places Conclusion. To prove that the primers you use can distinguish Ganoderma well, you need to test them on several Ganoderma species of known species. So, please go only to your finding. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? Partly Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Partly Competing Interests No competing interests were disclosed. Reviewer Expertise Phytopathology, Mycology, Seed Pathology, Biological Control I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (1) Author Response 11 Sep 2025 Emmanuellah Lekete-Lawson, Dep. of Microbiology, Czech University of Life Sciences Prague, Prague, Czech Republic Response to reviewer’s comment 2- Reviewer: Robert Paterson, Centre for Biological Engineering, University of Minho, Braga, Portugal Comment on Lekete-Lawson et al. (2025) By: Robert Russell Monteith Paterson University of Minho, Portugal [email protected] #My first concern when reading Lekete-Lawson et al. (2025) was the PCR analysis and subsequently I detected other problems. PCR Comment1: In relation to the PCR, in the text it says Figure 7A is for the culture based samples but these are Figure 7B in the legend to the image of the gel. Figure 7 B in the text is for basidiocarps but these are 7A in the legend to the figure. Which is it? Response:1 It was a typing mistake and has been corrected see page 14, Figure 7. Comment 2. Also, there is no information about the strain numbers that represented tracks 1 to 24 in the figure of the gels. These should be provided. It is all very confused. Also, there appeared to be a faint band in the blanks for samples 1 to 24 which were at the same intensity for track 20. Presumably the blanks were contaminated. Were tracks 20 and 35 from the same specimen and strain? Response 2. Yes, they are from the same sample but cannot confirm the strain, identity until confirm molecularly, this is because we were dealing with several new strains the faintness could probably due to either contamination from the DNA extraction or poor gel loading or not what we were expecting. b . The strain number could be captured in the gel because the software we were using was not programmed in that order, however detail of the individual strain numbers have been provided in the datataset repository. Comment 3: Was there a procedure for identifying inhibition of the PCR? I know that an internal amplification control (IAC) to assess inhibition is recommended for at least some PCRs. IAC would provide information on inhibition or could it be that the samples represented by 20 and 35 were not what you thought they were? Response.3a. We first suspected inhibitors like EDTA/Salts or even phenol after thoroughly purification DNA products; we still had the same results, probably not what we were expecting. Why did you not run known relevant Ganoderma species for comparison with the other specimens? This would provide information on the novelty of your specimens. Overall, there are considerable problems with the presentation of the PCR results. Response 3b . We were running what was available, we tried to identify and confirm if what we have in our country could be the same elsewhere. Having gotten the sequence data (Accession numbers) we then compared them with the known and existing ones in the data base, which helped us confirm our findings. Other concerns are as follows: General A strain and specimen list is necessary with accession numbers. Response: Strain and accession numbers have been provided in the write-up, together with phylogenetic trees giving a detailed explanation. Moreover, other details explanations/documents can be found in the figshare repository Comment 4 Are you saying G. ryvardenii is the only species involved in BSR in Ghana? What is the new species you say you have discovered? This is unclear. Response 4: Please I have stated it with percentage occurrence of the Ryvadenii and other species identified, however, The occurrence of G. ryvadenii was higher 83% compared to the remaining ones identified, even there was 1% G. woerese which has never been identified anywhere hence was discarded. So in total four different Ganoderma species were identified including the famous G. boninnense . But there percentages were very minimal. Comment: A considerable amount of this paper is taken from a previous paper (Lekete-Lawson et al. 2024). This paper should be discussed in the introduction and cited properly. And any duplication needs to be removed. Response: The study is ongoing with three main objectives, each autonomous but with a common goal of solving a disease problem. For each objective achieved, we release a publication. There was no repetition of the writing; rather, we showed how one objective is linked to another by adopting some of the procedures used in a previous to achieve the current objective and referencing it appropriately, it is not repetition but continuation and updates. Comment: Somebody who is excellent at writing scientific papers in English should be asked to rewrite the paper. Response: Well noted Abstract Comment: Palm oil is not necessarily the foremost vegetable oil. For example, soy oil is arguably more important. Response: This is very true, but Several studies (Breton et al, 2006; Bharudan et al., 2022) have proven that oil palm is a naturally and high-yielding oil crop,13 times more than soy and any other oil crops combined. Oil palm does not need any artificial additives to generate its oil. Comment: The abstract is vague??? What do you mean by using nucleic acid as a template? Response: If we said we used nucleic acid as a template, it means we used DNA as the starting material in a molecular assay to amplify the genetic sequences. For example, we extracted DNA from the culture-based isolates and that of the Basidiocarps and in this context, nucleic acid as a template means DNA was used as an input material to generate measurable results in the experiment. Comment: The English is poor, e.g. “BSR is characterised by stem decay, large-perennial, woody brackets basidiocarps”. What does this mean? Response: This is rewritten as: “Basal Stem Rot (BSR) is characterised by progressive stem decay coupled with the formation of large, perennial, woody basidiocarps with the average measurement of 2-65 cm in diameter on infected palms”. Comment: The conclusion does not make sense to me at all. Response: Conclusion has been rewritten “This study presents the first report of Ganoderma ryvardenii causing BSR disease on oil palm in Ghana, potentially the first in West Africa, and second in Africa. Notably, the pathogen was previously first reported to cause similar disease on oil palm in Cameroon, highlighting its emerging threat to oil palm production in the Sub-Saharan African region”(See page 2 of the manuscript) Introduction India is only a small palm oil-producing country in contradiction to what you have written. Respponse: Okay, well noted. Comment: You say Africa produces 3% oil palm, and then West Africa produces 4%. Which is it? Response: Africa accounts for approximately 3% of global palm oil production, with West Africa contributing about 4% of the continent’s total output. This implies that West Africa contributes 4% of Africa’s 3% share of global production. For example, if Africa produces 100,000 metric tonnes of palm oil, representing 3% of the global total, then West Africa contributes approximately 4,000 metric tonnes." Comment: G. boninense is the most serious disease of oil palm in SE Asia and Oceania. In Africa, it is considered that Fusarium wilt is the most serious fungal disease. Response: Contrary to your assertion in Ghana, research has shown that Fusarium is no longer a serious disease in our plantation and certainly not Ganoderma too. the case about Ganoderma is an emergent one, though reported many decades ago by Turner (1967), but was regarded as not economically important till now. Comment: G. boninense is accepted as the fungus responsible for BSR in SE Asia at least. You need to mention that G. ryvardenii was identified in Cameroon before and give the citation. Response: This was corrected in the introduction (See page 3) Materials and Methods Figure 1 should be in the results. Response: That was how it was structured, in the original manuscript, but changed by the editor Comment: In the sampling and isolation section, what does “including fruiting bodies (Basidiocarps) and Ganoderma sp. (Dikaryotic)” mean? Response: This was rewritten for better clarity as: “A total of ninety (90) symptomatic samples comprising basidiocarps and infected rotten plant tissues were collected from oil palms across the three agro-ecological regions under investigation. Comment: How were specimens tested for purity? Response: This was done with the combination of visual inspection and DNA-based confirmation (advanced purity check) using Universal (ITS) primers, thus a single clear band is likely considered pure. Comment: What does “Microzone, UK, proprietary formulation microLYSIS®-PLUS (MLP) underwent rapid thermal cycling for cell lysis and release of DNA” mean? Response: For DNA extraction, a proprietary formulation, microLYSIS ® -PLUS (MLP) from Microzone, UK, was used, for rapid thermal cycling to ensure efficient cell lysis and DNA release. Comment: GanET is G. boninense specific (or better, a palm clade specific) primer. It is not basidiomycete-specific. Please correct this. Response: Well noted, however, according to Mei Lieng Lo (2023) and Mandal et al., (2014), GanET is a DNA primer specifically designed for detection of Ganoderma boninense from oil palm tissue. Their study highlights the effectiveness of this primer in early pathogen detection even before visible symptoms appear. Results Comment: Did you test the difference between upper stem rot specimens and basal stem rot specimens? These have been considered as different species before. Response: Yes, we did, and it is still under investigation. This is because we identified it on only one palm at one spot, so we put the entire plantation and the area under surveillance, looking for more evidence to justify its discovery. Upper Stem Rot is known to be caused by Ganoderma Zonatum , which is related to G. boninense . In our study, we did not identify G. zonatum, but rather we had 7 % matching G. wiiroense and 4 % matches G. boninense. Nonetheless, G. wiiroense appeared to be a new strain/ species which has never been identified or published before, hence placed under further investigation. Comment: Phytophthora is not considered to be a fungus. It is an oomycete. Response: Well noted The Phylogenetic results section is poor with inadequate information. Response : More detailed explanation has been added. Additional data were provided in the figshare repository. (Figshare repository and can be accessed at [DOI: 10.6084/m9.figshare.29382632). Discussion Comment: Cameroon is not in West Africa. Response: Cameroon is in Central Africa, not in West Africa, but it is part of Sub-Saharan African countries, including Ghana. View more View less Competing Interests No competing interest reply Respond Report a concern Tondok ET. Peer Review Report For: First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved, 2 approved with reservations] . F1000Research 2025, 14 :413 ( https://doi.org/10.5256/f1000research.178088.r380559) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/14-413/v1#referee-response-380559 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2025 Supriyanto S. This is an open access peer review report 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. 30 Apr 2025 | for Version 1 Supriyanto Supriyanto , Tanjungpura University, Jl. Prof. Dr. H. Hadari Nawawi, Pontianak, Indonesia 0 Views copyright © 2025 Supriyanto S. This is an open access peer review report 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. format_quote Cite this report speaker_notes Responses (1) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Overall, this article is well written. However, some parts need to be strengthened to make it a reliable reference. 1. In the last paragraph of the introduction, considering the author's claim about the discovery of a new species of Ganoderma that causes Basal Stem Rot disease in oil palm, it is better to add a new, more complete reference about reports related to the types of Ganoderma that previous researchers have reported. 2. Method. Observation of disease severity in oil palm in the field uses a score of 0 to 4. The scoring criteria should be displayed in full. In addition, the use of the number of fruiting bodies as an indicator of severity seems rather risky because there has been no report on this. Based on experience in Indonesia, the number of fruiting bodies that can be found is not directly related to the severity of the disease. 3. Method, pathogenicity test. To be clearer, it is necessary to further describe how to arrange RWB, oil palm seeds, and the media used in the container. Is RWb mixed with the media, or how? 4. Results, point 3.2. This subtitle is a bit inappropriate, because the content is a description of the morphology of the Ganoderma fruit body found. It would be better if the title were "Morphological characteristic......" 5. Results, Figure 2. In part (B), it is stated as the Initial stage of infection and the pinhead of the Ganoderma mushroom. This is a less appropriate statement, because the initial infection of Ganoderma on oil palm is not related to the appearance of pinheads. The appearance of pinheads is a sign that the infection is advanced and the disease is severe. 6. Figure 5. The image is not clear. The 0.5 mm line mark does not seem to match the original condition. In part (ii), root decay, necrosis, and decaying bole tissue are not visible. If possible, the image can be displayed at a closer up, so that the message to be conveyed can be better received by the reader. 7. Table 2. If possible, it would be better if the disease severity data could be displayed. 8. Figure 6. Not necessary. This image shows nothing. 9. It would be better if a general discussion were added that supports the main findings. For example, the condition of the environmental vegetation around the sample plantation is suspected of causing G. ryvardenii to be there and attack oil palms. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? Partly Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise Plantation crop cultivation, plant disease, oil palm disease, and biological control of plant disease. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (1) Author Response 11 Sep 2025 Emmanuellah Lekete-Lawson, Dep. of Microbiology, Czech University of Life Sciences Prague, Prague, Czech Republic Response to Reviewer’s comment Reviewer 1 Reviewer 1: Overall, this article is well written. However, some parts need to be strengthened to make it a reliable reference. Comment 1. In the last paragraph of the introduction, considering the author's claim about the discovery of a new species of Ganoderma that causes Basal Stem Rot disease in oil palm, it is better to add a new, more complete reference about reports related to the types of Ganoderma that previous researchers have reported. Response1: If I understand your comment, I believe you want us to elaborate on the discovery work conducted by previous researchers concerning the species identified in the current study. If so, here is the reference to the work done by: Researchers (Kinge TR and Mih AM), In 2010 and 2011 these researchers conducted a comprehensive study in the littoral and southwestern regions of Cameroon to identify Ganoderma species associated with basal stem rot (BSR) disease of oil palm. During the survey, these researchers encountered a previously unclassified Ganoderma species, which morphological and molecular analyses revealed to be distinct from all known species ever reported. To determine its identity, a morphological and molecular characterisation of the ITS rDNA locus was performed, which revealed variation in ellipsoidal basidiospores with narrowly truncate apices, thereby differentiating it from its close relatives like G. steyaertanum and the G. boninense clade, but it stands as a strongly supported standalone lineage. Expressing their admiration for its distinctiveness, the researchers named it Ganoderma ryvardense, in appreciation of the extraordinary mycologist Leif Ryvarden, whose outstanding work in African fungal taxonomy cannot be ignored. (This is inserted on page 3 of the revised manuscript.) Comment-2. Method . Observation of disease severity in oil palm in the field uses a score of 0 to 4 . The scoring criteria should be displayed in full . In addition , the use of the number of fruiting bodies as an indicator of severity seems rather risky because there has been no report on this. Based on experience in Indonesia, the number of fruiting bodies that can be found is not directly related to the severity of the disease. Response 2a: The Disease Scoring Criteria is displayed on page 5 of the manuscript as seen in the table below: Disease severity score Disease severity was scored using a scale of 0 to 4 (Lekete-Lawson et al. , (2024); Guide Disease severity index for Ganoderma infection (BSR) of oil palm Severity score No. of BSR fruiting bodies/foliar symptoms /plant Percent diseased palm 0 No BSR fruiting body/no foliar symptoms Healthy plant or no disease 1 1– 3 BSR fruiting body/foliar symptoms on two fronds 1– 25 % 2 4– 6 (BSR) fruiting body/foliar symptoms on three fronds 26– 50% 3 7– 9 (BSR) fruiting body/foliar symptoms on four fronds 51– 75% 4 > 9 (BSR) fruiting body/foliar symptoms on > four fronds 76 –100% Response 2b. Reasons of using fruiting bodies as an indicator of severity: The use of fruiting bodies as a severity indicator was adopted for the purpose of this study and was derived from the idea in the mushroom production technique. Ganoderma pathogen is a white rot fungal disease and difficult to diagnose at early stage and can also be misdiagnosed if using only foliar symptoms of the infected plant alone, less you see the mushroom which is the sign depicting the surest presence of the disease. Disease severity measures how badly the plant is affected, not just whether it is infected or how many of the plants are infected (incidence). The amount of mushrooms present per spot gives us an idea of how severely a plant is damaged as a result of the infection and the presence of the inoculum load. Also, disease severity focuses on the Intensity of the damage (e.g., % of area covered with lesions) or the percentage plant part (stem) covered with the mushroom; hence, the amount of mushroom coming out from the stem gives us an idea of how severely that plant is infected. How intense is the infection? This is the first time an intensive study has been conducted since the outbreak of Ganoderma Basal stem Rot disease in Ghana; we do not have sophisticated instruments/tools to detect how severely the plant is affected than to use the visual sign of the disease to show. Comment 3: Method : pathogenicity test. To be clearer, it is necessary to further describe how to arrange RWB, oil palm seeds, and the media used in the container. Is RWb mixed with the media, or how ? Response 3: In this experiment, we have stated that due to the scarcity of the Rubber Wood Block (RWB), we improvised RWB with Wawa wood block (WWB), not RWB. We adopted techniques by Angel et al. (2021) and Bivi et al., ( 2016 ), and modified to suit our study and in each treatment bottle, the mixture of the growing media in the bottle were also clearly stated under the methodology. (A volume of 250 ml Beatson glass jars (R3/83 mm 4 Doz) containing a mixture of planting media (black soil, bio-cha, and treated plant biostimulant) at the ratio of 2:2:1 100 g per glass was used. The jars were autoclaved for 45 minutes at 121°C. The mixtures were left to cool for three days) Comment 4 : Results , point 3.2. This subtitle is a bit inappropriate, because the content is a description of the morphology of the Ganoderma fruit body found. It would be better if the title were " Morphological characteristic......" Response 4: The title Physiological characteristics is replaced with morphological characteristics as suggested on page 8. Thanks. Comment 5. Results, Figure 2. In part (B), it is stated as the Initial stage of infection and the pinhead of the Ganoderma mushroom. This is a less appropriate statement, because the initial infection of Ganoderma on oil palm is not related to the appearance of pinheads. The appearance of pinheads is a sign that the infection is advanced and the disease is severe. Response 5: The pinhead is the first visible sign of the mushroom or the initial sign of the infection. Basal stem rot disease is often accompanied by the formation of the fruiting bodies on the lower/upper trunk and exposed root area as depicted in the picture (Fig. 2), In our case foliar symptoms were not prominent hence the appearance of the mushroom (pinhead was the only indication of the presence of the pathogen showing initial sign of the disease) This could probably be unique with this particular species “no foliar symptom but presence of the mushroom”. Comment 6. Figure 5. The image is not clear. The 0.5 mm line mark does not seem to match the original condition . In part (ii), root decay, necrosis, and decaying bole tissue are not visible. If possible, the image can be displayed closer up, so that the reader can better receive the message to be conveyed. Res: 6 The image has been replaced on page 11. However, the experiment was done on the two-week-old germinated seed nuts; hence, the bole of the seed nut was not visibly pronounced as compared to if the experiment were to be done on one or two-month-old seedlings. Comment 7 . Table 2. If possible, it would be better if the disease severity data could be displayed. Response 7: The disease severity data was provided in the raw dataset repository at (figshare.com repository: Lekete-Lawson E: Report on Basal Stem Rot (BSR) disease on oil palm in Ghana. figshare. Journal contribution. 2025b. 10.6084/m9.figshare.28329830.v1); for further explanation and understanding, however, to avoid overcrowding we chose to present the ANOVA table in the results section. Comment 8 . Figure 6. Not necessary. This image shows nothing. Response 8: Figure 6 is removed (page 12) Comment 9. It would be better if a general discussion were added that supports the main findings. For example, the environmental vegetation around the sample plantation is suspected of causing G. ryvardenii to be there and attack oil palms. Response 9: This is the initial study and the first report; the study is ongoing, although there have never been any report of G. ryvadenii associated with the type vegetation of vegetation we have or any tree crops or oil palm in Ghana in the past, however subsequent studies into other factors including the environmental and host factors that could lead to the presence of G. ryvadenii is ongoing. Meanwhile, the climatic conditions of the individual study areas are well stated under the climate of the study areas in the methodology Page 4. View more View less Competing Interests Authors declare no competing interest reply Respond Report a concern Supriyanto S. Peer Review Report For: First Report of Ganoderma ryvardenii causing Basal Stem Rot (BSR) disease on oil palm ( Elaeis guineensis Jacq.) in Ghana [version 1; peer review: 1 approved, 2 approved with reservations] . F1000Research 2025, 14 :413 ( https://doi.org/10.5256/f1000research.178088.r377597) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/14-413/v1#referee-response-377597 Alongside their report, reviewers assign a status to the article: Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. 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