Mor. J. Agri. Sci. 2 (1): 9-13, March 2021 9 Inocybaceae and affiliated taxa from West Africa Hyppolite L. AIGNON1*, Arooj NASEER2, Azize A. BOUKARY1, Nourou S. YOROU1 Abstract 1 Research Unit Tropical Mycology and Plant-Soil Fungi Interactions, Faculty of Agronomy, University of Parakou, Benin 2 Department of Botany, University of the Punjab, Quaid-e-Azam Campus, Lahore, Pakistan * Corresponding author hyppoliteaignon@yahoo.com Received 26/01/2021 Accepted 24/02/2021 Inocybaceae and its affiliated taxa recorded in West Africa were examined by bibliographic survey through available publications coupled with field data collections in Benin, Burkina Faso, Côte d’Ivoire, Guinea, Mali and Togo. Twenty-one Inocybaceae and affiliated taxa have been reported in the region. All taxonomic names have been checked in Index Fungorum and the distribution of Inocybe beninensis, I. flavipes, I. fuscobrunnea, I. pallidiangulata, Inosperma africanum, I. bulbomarginatum, I. flavobrunneum, Mallocybe africana, Pseudosperma afrofibrosum, P. beninense and P. fragilipes in West Africa were reported. In addition, results of Basic Local Alignment Search Tool search including some undescribed collections from Zambia, such as PC96082, PC 96204 with PC:0088767 and L4512 Inoc Zam05 that showed similarity respectively with the collections of Inosperma bulbomarginatum, Mallocybe africana and Inocybe flavipes from West Africa with >97% similarity. Keywords: Checklist, Ectomycorrhizal fungi, Inocybe, Distribution, Pseudosperma, West Africa INTRODUCTION Inocybaceae Julich, is monophyletic family with a worldwide distribution. It encompasses nearly 1,050 species (Matheny et al., 2020) found mainly in forests (from woodland, gallery or dense forests) and rarely in grasslands, wetlands or agricultural lands (Ov, 2015). The species of Inocybaceae are ectomycorrhizal and associate with 23 families of vascular plants (Matheny et al., 2020). Recently, Matheny et al. (2020) suggested a revised phylogeny for the family Inocybaceae. Today, Inocybaceae contains seven genera, Auritella Matheny & Bougher, Inocybe (Fr.) Fr., Inosperma (Kühner) Matheny & Esteve-Rav., Mallocybe (Kuyper) Matheny, Vizzini & Esteve-Rav, Nothocybe Matheny & K.P.D. Latha, Pseudosperma Matheny & Esteve-Rav and Tubariomyces Esteve-Rav. & Matheny (Matheny et al., 2020). However, due to similarity and lack of molecular analyses, a large number of Inocybaceae taxa were classified in Cortinariaceae Singer (Horak, 1978; Singer, 1986, 1975) and probably all that were recognized then, which may include taxa that are not recognized now such as Astrosporina Schröt. In the literature, most of the premolecular classifications merged not only Inocybaceae with affiliated taxa, Crepidotaceae Singer and Tubariaceae Vizzini. The first taxonomic evaluation within Inocybaceae was based on morphological and microscopic characters for the distinction of genera earlier (Kuyper, 1986). Beforehand, Jülich (1982) placed Inocybe (Fr.) Fr. in Inocybaceae together with the genus Astrosporina which is now known to be polyphyletic and nested within Inocybe (Matheny, 2005; Matheny et al., 2002). Except some scarce publications (Aïgnon et al., 2021a; b; Buyck and Eyssartier, 1999; Matheny and Watling, 2004; Piepenbring et al., 2020) taxonomic documenta© Moroccan Journal of Agricultural Sciences • e-ISSN: 2550-553X tion of the family Inocybaceae from tropical Africa is scant. But, recent mycological sampling in West Africa reported eleven new species to science that some of which like Inosperma africanum, I. bulbomarginatum, I. flavobrunneum and Mallocybe africana have been published (Aïgnon et al., 2021a;b). Though taxonomic works are still progressing, it is of paramount importance to provide a state-of-the-art related to the diversity known in West Africa, in order to establish a reference guide to detect new species. In this paper, we provide a checklist of known species of Inocybaceae and affiliated taxa from West Africa based on the literature, observation and field data collection between 2013 and 2018. MATERIALS AND METHODS Study area West Africa is defined here to include 15 countries (Benin, Burkina Faso, Côte d’Ivoire, Cape Verde, Gambia, Ghana, Guinea, Guinea-Bissau, Liberia, Mali, Niger, Nigeria, Senegal, Sierra Leone and Togo). The recently undertaken sampling trips and collection sites in Benin, Burkina Faso, Côte d’Ivoire, Guinea, Mali and Togo are indicated in Figure 1. Data compilation The data were assembled from literature related to Inocybaceae from West Africa. All scientific names have been cross-checked against Index Fungorum (IF) and synonymous names have been separated. Data from literature were coupled with field observations. We made field surveys from 2013 to 2018. Specimens were collected in Benin (Wari Maro forest www.techagro.org Aignon et al.: Inocybaceae from West Africa 10 reserve, Ouémé Supérieur forest reserve, Okpara forest and Toui-Kilibo forest reserve). Additional surveys were carried out in Burkina Faso (Toussianbandougou gallery forest, Dan gallery forest and Niangoloko forest reserve), Mali (Farako forest reserve), Guinea (Kouraouletediene forest reserve, Levari forest, Baroforest reserve, Moussaya forest reserve, Haut Niger National Park, Telaya forest, Tindo forest and Côte d’Ivoire (Kekrekouakoukro forest and Kouadianikro forest). The sampling sites are presented in Figure 1. DNA extraction, PCR and sequencing On the recent collection of Inocybaceae from Benin, Burkina Faso, Côte d’Ivoire, Guinea, Mali and Togo, DNA was extracted from dried badisiomata using a QIAGEN® plant mini kit and for the Polymerase chain reaction (PCR) amplification, three nuclear gene regions such as Internal Transcribed Spacer (ITS), portions of the large subunit ribosomal DNA gene (28S) and RNA polymerase II subunit (RPB2), were amplified using the following primers, ITS1F and ITS4 for the ITS region (Gardes and Bruns, 1993; White et al., 1990), LR0R, LR7, LR5 and LR3R for 28S (Cubeta et al., 1991; Rehner and Samuels 1995 Vilgalys and Hester, 1990,) and bRPB2-6F, bRPB2-7.1R for the most variable region of RPB2 (Matheny 2005). PCR products were cleaned and sequenced at Macrogen Inc. (Macrogen Europe B.V., Amsterdam, Netherlands) using the same primers as those used for PCR. All sequences of the new species are deposited on GenBank (Aïgnon et al., 2021a;b) and the new sequences were subjected to a BLAST search (97% similarity) and relevant related sequences retrieved from GenBank. RESULTS Inocybaceae and affiliated/Allied taxa in west Africa Twenty-one Inocybaceae and affiliated taxa have been reported from West Africa. Table 1 summarizes the different species of Inocybaceae as well as the affiliated taxa of West Africa. Existing data show that Inocybaceae and affiliated taxa have been recorded in four countries from West Africa i.e., Ghana, Nigeria, Senegal and Sierra Leone and collected in forests dominated by Adenia lobata Eng, Afzelia africana Sm. ex Pers., Afzelia bella Harms, Afzelia bracteata T. Vogel ex Benth, Anthonotha crassifolia Baill. and Uapaca chevalieri Beille. But, recent sampling in Benin, Burkina Faso, Côte d’Ivoire, Guinea, Mali and Togo has shown that the species of Inocybaceae are found in the vegetations dominated by Isoberlinia doka Craib & Stapf, I. tomentosa (Harms) Craib & Stapf, Uapaca togoensis Pax, U. guineensis Müll. Arg., Monotes kerstingii Gilg and Berlinia grandiflora Hutch. & Dalziel. At the 97% threshold, after BLAST searched at NCBI (www.blast.ncbi.nlm.nih.org), Inocybe sp. PC96082 from Zambia present high similarity with the collections Côte d’Ivoire Figure 1: Map of West Africa showing the studied areas and recent sampling sites Mor. J. Agri. Sci. 2 (1): 9-13, March 2021 11 of Inosperma bulbomarginatum, same for the collections PC 96204 and PC:0088767 from Zambia with the collections of Mallocybe africana from West Africa, as well as the collection L4512 Inoc Zam05 from Zambia with the collections of Inocybe flavipes from West Africa which appear to be the same species. DISCUSSION Uptill now, only five species of Inocybaceae, Inocybe ghanaensis (Pegler 1969; Piepenbring et al., 2020), Mallocybe africana (Aïgnon et al., 2021b), Inosperma africanum, I. bulbomarginatum, and I. flavobrunneum (Aïgnon et al., 2021a) are reported and well described from West Africa to our knowledge, but many taxa are still without description and wait to be published Inocybe beninensis, I. flavipes, I. fuscobrunnea, I. pallidiangulata, Pseudosperma afrofibrosum, P. beninense, P. fragilipes. However, some name is not validated such as Inocybe sp. “gbadjii” registered in index fungorum but it is not consequently described due to lack of diagnosis and type (Boa, 2004). In addition to these species there are many undescribed collections described as Inocybe spp. from Burkina-Faso, Guinea, Nigeria and Senegal (Redhead, 1968; Thoen and Ducousso, 1989; Bâ et al., 2012; Piepenbring et al., 2020). Recent findings (Aïgnon et al., 2021a;b) has increased the diversity of Inocybaceae in West Africa to more than eighteen species (Table 1). However, the Central and East Africa regions contain Table 1: Checklist of Inocybaceae and affiliated taxa from West Africa Family Species Crepidotus applanatus Authority (Pers.) P. Kumm. Host Plant Distribution Sierra Leone Field observation: Collected from dead trunks of Adenia lobata (Beeli, 1938) - Sierra Leone - (Holden, 1970) Nigeria Dead decaying wood (Tree stump and fallen logs), Decay trees, Dead decaying wood (Tree stump and fallen logs) (Beeli, 1938) (Osemwegie and Okhuoya, 2011) (Osemwegie et al., 2006) (Osemwegie et al., 2010) Benin Field observations: on soil In press Crepidotus mollis (Schaeff.) Staude - Inocybe beninensis Aïgnon, Yorou & Isoberlinia doka Ryberg and I. tomentosa I. flavipes Aïgnon, Yorou & Isoberlinia doka and Benin, Togo Ryberg I. tomentosa Field observations: on soil In press I. fuscobrunnea Aïgnon, Yorou & Burkina Faso, Berlinia grandiflora Côte d’Ivoire Ryberg In press I. ghanaensis Pegler Field observations: on soil Field observations: on bare soil Field observations: on soil - (Boa, 2004) - Ghana Burkina Faso, I. pallidiangulata Aïgnon, Yorou & Berlinia grandiflora Côte d’Ivoire Ryberg Inocybaceae References Adenia lobata Crepidotaceae Crepidotus caspari Velen. Observations Inocybe sp. - Benin Inocybe sp. Afzelia africana Senegal Inocybe sp. Afzelia bella Senegal Inocybe sp. Anthonotha crassifolia, Uapaca chevalieri Senegal Inocybe sp. Afzelia africana Senegal Inocybe sp. Afzelia africana Senegal Burkina Aïgnon, Yorou & Isoberlinia doka and Benin, Faso, Guinea, Ryberg Berlina grandiflora. Côte d’Ivoire, Togo Aïgnon, Yorou & Isoberlinia doka, I. Benin, I. bulbomarginatum Ryberg tomentosa Aïgnon, Yorou & Isoberlinia doka and Benin, I. flavobrunneum Ryberg I. tomentosa Togo, Aïgnon, Yorou & Isoberlinia doka and Benin, Mallocybe Burkina Faso, Ryberg africana I. tomentosa Côte d’Ivoire Inosperma africanum P. afrofibrosum P. beninense P. fragilipes Aïgnon, Yorou & Isoberlina doka Ryberg Aïgnon, Yorou & Isoberlina doka Ryberg Aïgnon, Yorou & Isoberlina doka Ryberg Benin Benin Benin (Pegler, 1969) (Holden, 1970) In press Field observations: Appeared on lateral (Bâ et al., 2012) roots of Afzelia africana Field observation: Appeared near of (Redhead, 1968) Afzelia bella Test synthesis mycorrhizae in vitro or in semi-axenic condition Rootlets fixation in formaldehyde-acetic acid Rootlets fixation in formaldehyde-acetic acid Field observations: on soil Field observations: on soil Field observations: on soil Field observations: on soil Field observations: on soil Field observations: on soil Field observations: on soil (Thoen and Ducousso, 1989) (Thoen and Bâ, 1989) (Thoen and Bâ, 1989) (Aïgnon et al., 2021a) (Aïgnon et al., 2021a) (Aïgnon et al., 2021a) (Aïgnon et al., 2021b) In press In press In press Aignon et al.: Inocybaceae from West Africa 12 high diversity of Inocybaceae. The diversity of Inocybaceae is also high in the Central and East Africa regions where Auritella encompasses three species such as Auritella hispida Matheny & T.W. Henkel, Auritella spiculosa Matheny & T.W. Henkel and Auritella aureoplumosa (Watling) Matheny & Bougher from Cameroon, as well as two undescribed species Inocybe sp. TU112047 and Inocybe sp. TU112061 from Gabon (Matheny et al., 2017, 2012). In Tubariomyces, there is an undescribed species Tubariomyces sp. 2 BB6018 (PC), from Zambia (Vizzini et al., 2013). Inosperma misakaense (Matheny & Watling) Matheny & Esteve-Rav. is known from Zambia (Aïgnon et al., 2021; Buyck and Eyssartier, 1999; Gardens, 2017; Matheny and Watling, 2004). As well as, there are several undescribed species from Zambia including Inocybe sp. PC 96042, Inocybe sp. PC 96039, Inocybe sp. PC 96081, Inocybe sp. PC 96095, Inocybe sp. PC 96204, Inocybe sp. PC 96111, Inocybe sp. PC 96013, Inocybe sp. PC 96083, Inocybe sp. BB3233, Inocybe sp. BB6018 and Inocybe sp. PC 96073 (Matheny et al., 2009). Ectomycorrhizal symbiosis is proven between some species of Inocybaceae and the roots of native trees. For example, Anthonotha crassifolia and Uapaca chevalieri are partner trees of some unidentified Inocybaceae species in the semi-deciduous forest of Senegal (Thoen and Ducousso, 1989), likewise for Afzelia bella in the tropical forest of Nigeria (Redhead, 1968) and also Afzelia africana in wooded areas and gallery forest in Senegal (Thoen and Bâ, 1989). In addition, these ectomycorrhizal host trees are found in various vegetations such as Zambesian and Sudanian woodlands, semi-deciduous forests, Guineo-Congolean dense forests and gallery forests. The presence of Inocybaceae species in these vegetations is remarkable, and there is little doubt that other African species of Afzelia, Anthonatha and Uapaca are also ectomycorrhizal associated with Inocybe but it is difficult to confirm without consequent analysis. Analysis of the recent collections (Aïgnon et al., 2021a,b) have shown a wide distribution of Inocybaceae species in vegetations dominated by Uapaca spp., Isoberlinia spp. and Berlinia grandiflora with large distributed in Benin, Burkina Faso, Guinea, Senegal and Togo (Thoen and Ducousso, 1989; Moyersoen and Fitter, 1999; Newbery and Stoll, 2013). The same is true for Monotes kerstingii often mixed with stands of Isoberlinia spp. (Sanon et al., 1997). It is then clear that Inocybaceae have not been sufficiently evaluated in West Africa and so there is need to study the biology and distribution of these species to determine specific host trees for conservation interventions. Acknowledgements We are highly indebted to National Geographic Society (grant No. CP 126R-17) for financial assistance for collection of many of the specimens analyzed in this paper. 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