The Genus Ravenelia (Pucciniales) in South Africa
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Mycological Progress (2020) 19:259–290 https://doi.org/10.1007/s11557-020-01556-w ORIGINAL ARTICLE The genus Ravenelia (Pucciniales) in South Africa Malte Ebinghaus1 & M. J. Wingfield2 & D. Begerow1 & W. Maier3 Received: 23 August 2019 /Revised: 30 December 2019 /Accepted: 2 January 2020 # The Author(s) 2020 Abstract The genus Ravenelia represents the third largest genus of rust fungi and parasitizes a great number of leguminous shrubs and trees, mainly in the subtropics and tropics. Molecular phylogenetic analyses of this genus using nc 28S rDNA and CO3 sequences are presented with a special focus on South African representatives of Ravenelia. Many of the specimens had been collected by us in recent years, mainly from acacia species of the genera Vachellia and Senegalia. Morphological characters were extensively studied using light microscopy and scanning electron microscopy. The analyses resolved several well-supported phylogenetic groups. By linking these groups to their morphology and life cycle characteristics, it was possible to interpret the outcomes in terms of their evolutionary ecology and biogeography. Several characters previously used to define subgeneric groups within Ravenelia were found to be misleading because of assumed convergent evolution. However, host associations, the ability to induce aecial galls as well as the development of two-layered probasidial cells emerged as useful criteria for inferring mono- phyletic groups. Six novel Ravenelia species were discovered and described. Furthermore, five species represent new reports for South Africa, species descriptions were emended for two taxa, and a new host report emerged for R. inornata. Keywords Pucciniales . Raveneliaceae . Ravenelia moloto sp. nov. Ravenelia molopa sp. nov. Ravenelia modjadji sp. nov . Ravenelia doidgeae sp. nov. Ravenelia spinifera sp. nov. Ravenelia dumeti sp. nov. Phylogeny . Diversity study Introduction Sphaeria epiphylla Schwein. collected on Tephrosia virginiana (L.) Pers. in South Carolina and the newly described R. indica In 1853,BerkeleyintroducedthegenusRavenelia within the rust Berk. found on pods of an unidentified acacia. Later then Dietel fungi (Pucciniales). The genus initially comprised two species: correctly recombined the type species R. glandulosa to R. R. glandulosa Berk. & M.A. Curtis, which was transferred from epiphylla (Schwein.) Dietel (Dietel 1894). In subsequent years, many additional Ravenelia species were found throughout the tropics and subtropics and today, some 200 species are described This article is part of the topical collection on Basidiomycote Mycology in (Hernández and Hennen 2002,CumminsandHiratsuka2003). honor of Franz Oberwinkler who passed away in March 2018 Ravenelia thus became the third most species-rich rust fungal Section Editor: Meike Piepenbring genus after Puccinia and Uromyces. All known species of Ravenelia are confined to a great diver- Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11557-020-01556-w) contains supplementary sity of Fabaceae residing in all three traditionally recognized material, which is available to authorized users. subfamilies (Mimosoideae, Faboideae, Caesalpinioideae) (Hennen et al. 2005). The most prominent morphological fea- * Malte Ebinghaus tures that are shared by all species of Ravenelia are the multicel- [email protected] lular teliospores, which are borne on compound pedicels com- posed of two to several hyphae. These spores have an ellipsoidal, 1 AG Geobotanik, Ruhr-Universität Bochum, Bochum, Germany reniform, or almost hemispherical shape in side view and bear a 2 FABI Forestry and Agricultural Biotechnology Institute, University variable number of pendent hygroscopic cysts. Other characters of Pretoria, Pretoria, South Africa include spermogonia of type 5 and 7 (Cummins and Hiratsuka 3 Institute for Epidemiology and Pathogen Diagnostics, Federal 2003)butthesearecommonlyabsent. Research Centre for Cultivated Plants, Julius Kühn-Institut, All species of Ravenelia are autoecious and their life cycles Braunschweig, Germany range from macro- and demi to hemi-, and more rarely to 260 Mycol Progress (2020) 19:259–290 microcyclic (Cummins and Hiratsuka 2003). The aecial stage conflicts when applying the taxonomic system for of several macrocyclic Ravenelia spp. can easily be recog- Ravenelia proposed by Sydow (1921), we mapped his sug- nized in the field by their ability to induce galls and witches’ gested nomenclatural system to the phylogenetic recon- brooms in host tissues. Another special feature is the produc- structions and discussed these outcomes. tion of uredinoid aecia in numerous species of Ravenelia (Cummins 1959; Cummins and Hiratsuka 2003). The morphological diversity and the variability of Material and methods Ravenelia life cycles prompted mycologists early in the twen- tieth century to establish sections (Long 1903; Dietel 1906) or Specimens examined even to split this genus into several distinct genera (Sydow and Sydow 1915; Sydow 1921). The most sophisticated tax- The specimens used for the molecular phylogenetic and onomic system for Ravenelia was proposed by Sydow (1921) morphological analyses were collected during several field distinguishing eight genera based on teliospore traits in com- surveys from 2004 to 2015 in South Africa. In addition, we bination with observed life cycles. Details of the competing considered 13 herbarium specimens from BPI originating taxonomic systems are summarized in Table S1. However, a from North and South America as well as DNA sequences broad genus concept of Ravenelia comprising all these sug- downloaded from GenBank (see Table 1). Freshly collect- gested genera or sections (within one genus) remains most ed material was immediately dried between paper sheets in widely accepted (compare Cummins and Hiratsuka 2003). aplantpressanddepositedafterdeterminationatthe More than 500 rust species are known from South Africa National Collections of Fungi (PREM) in Roodeplaat, (Crous et al. 2006), making this country relatively well explored South Africa, and the herbarium of the Natural History for these fungi in comparison with other countries in Africa. Museum in Karlsruhe (KR), Germany. In total, 91 speci- Most contributions to the collection and description of rust fungi mens representing 44 Ravenelia species and three outgroup in South Africa are attributed to the investigations of Ethel M. species were included in the molecular phylogenetic anal- Doidge during the first half of the twentieth century. In her last yses and all of them were examined microscopically. For comprehensive species list of southern African rust fungi, she comparative purposes, additional 32 specimens comprising mentioned 24 Ravenelia species eight of which she described 15 type specimens deposited at PREM were examined only (Doidge 1927, 1939, 1950). The most recent species list was microscopically. All specimens investigated in this study published by van Reenen (1995)butnearlyexclusivelyrelied are listed in Table 1. on literature data provided by Doidge. Due to changes in political borders, two species each now only occur in Mozambique (R. Light- and electron microscopic investigations deformans and R. le-testui)andZimbabwe(R. indigoferae and R. bottomleyae)respectively,whileR. baumiana was recorded only The spores from plant material were scraped from the infected from Angola. Two rusts, R. atrides and R. bottomleyae,were tissues using sterile insect needles and mounted in lactophenol transferred to the genera Uredopeltis (Wood 2007)and solution on microscope slides. Light microscopic examina- Spumula (Thirumalachar 1946), respectively. Wood (2006)re- tions were made using a Zeiss Axioplan light microscope corded R. ornata for the first time in South Africa and Ebinghaus (Carl Zeiss Microscopy, Jena, Germany) with a Color View et al. (2018) described R. xanthophloeae on the Vachellia microscope camera (Olympus Soft Imaging System, Münster, xanthophloea.Thus,19Ravenelia species are currently known Germany) and a Zeiss Axio Imager M2 microscope with an for South Africa. Axiocam 506 camera (both Carl Zeiss Microscopy, Jena, During the course of recent surveys, aiming at re- Germany). Morphological characteristics were measured collecting the majority of Ravenelia species from South using Cell^D v. 3.1 imaging software (Olympus Soft Africa and especially at investigating all known Acacia Imaging Solutions GmbH, Münster, Germany) and Zen2 s.l. for potential rust infections, we have collected numer- Blue Edition v.2.3 (Carl Zeiss Microscopy GmbH, Jena, ous specimens from acacias and fabaceous plants. The Germany). The specimens comprising Ravenelia albizziicola overarching aims were to re-evaluate the species diversity (PREM40295), R. baumiana (PREM50553, PREM29870, and systematics of Ravenelia rusts in South Africa by PREM6886), R. elephantorhizae (PREM8955), R. using microscopic investigations and molecular phyloge- escharoides (PREM534), R. glabra (PREM2375, netic techniques. For a better understanding of the phylog- PREM10698), R. halsei (PREM30117, PREM50751), eny of the genus as a whole also species mainly from the R. inornata (PREM2368, PREM2541, PREM20734), Neotropics were investigated. The emerging phylogenetic R. minima (PREM30779, PREM10697), R. modesta clades were interpreted using aspects of biogeographical (PREM34572, PREM30110), R. natalensis (PREM2514, distributions, life cycle traits,andhostassociationsaswell PREM1935), R. peglerae (PREM2544, PREM5626, as morphological