Ophiostoma gemellus and Sporothrix variecibatus from mites infesting Protea infructescences in South Africa

Ophiostoma (Ophiostomatales) represents a large genus of fungi mainly known from associations with bark beetles (Curculionidae: Scolytinae) infesting conifers in the northern hemisphere. Few southern hemisphere native species are known, and the five species that consistently occur in the infructescences of Protea spp. in South Africa are ecologically unusual. Little is known about the vectors of Ophiostoma spp. from Protea infructescences, however recent studies have considered the possible role of insects and mites in the distribution of these exceptional fungi. In this study we describe a new species of Ophiostoma and a new Sporothrix spp. with affinities to Ophiostoma, both initially isolated from mites associated with Protea spp. They are described as Ophiostoma gemellus sp. nov. and Sporothrix variecibatus sp. nov. based on their morphology and comparisons of DNA sequence data of the 28S ribosomal, ß-tubulin and internal transcribed spacer (ITS1, 5.8S, ITS2) regions. DNA sequences of S. variecibatus were identical to those of a Sporothrix isolate obtained from Eucalyptus leaf litter in the same area in which S. variecibatus occurs in Protea infructescences. Results of this study add evidence to the view that mites are the vectors of Ophiostoma spp. that colonize Protea infructescences. They also show that DNA sequence comparisons are likely to reveal additional cryptic species of Ophiostoma in this unusual niche

Thecaphora capensis sp. nov., an unusual new anther smut on Oxalis in South Africa

The smut genus Thecaphora contains plant parasitic microfungi that typically infect very specific plant organs. In this study, we describe a new species of Thecaphora from Oxalis lanata var. rosea (Oxalidaceae) in the Cape Floristic Region of South Africa. Molecular phylogenetic reconstructions based on large subunit ribosomal DNA sequence data confirmed the generic placement of the fungus and confirmed that it represents an undescribed species for which the name T. capensis sp. nov. is provided. The closest known sister species of the new taxon is T. oxalidis that infects the fruits of Oxalis spp. in Europe, Asia and the Americas. In contrast, T. capensis produces teliospores within the anthers of its host. This is the first documented case of an anther-smut from an African species of Oxalis and the first Thecaphora species described from Africa

Genetic differentiation in Oxalis (Oxalidaceae): A tale of rarity and abundance in the Cape Floristic Region

AbstractOxalis L. is the largest and most diverse genus in the family Oxalidaceae. Within southern Africa, Oxalis is represented by ca. 270 taxa, the majority occurring in the Cape Region. Although many of the species are widespread, ca. 25% are considered rare. The aim of this paper is to assess the degree of genetic differentiation between two rare and highly localized species (Oxalis hygrophila Dreyer and Oxalis oligophylla Salter) and the more widespread Oxalis tomentosa L.f. For comparative purposes, we also include Oxalis purpurea L., one of the most widely distributed species in South Africa. Chloroplast sequences of the trnH/psbA spacer revealed low genetic diversity for O. oligophylla and O. tomentosa compared to the widespread O. purpurea. High genetic diversity in O. purpurea might, in combination with other ecological and reproductive factors, account for the success of this species. In contrast, low variation might contribute to rarity in O. oligophylla and ultimately ground O. tomentosa to become rare. The latter two species were not monophyletic with a shared haplotype. Coalescent modelling revealed low levels of gene flow (<1 migrant per generation) between them and we argue that the genetic pattern is the result of the retention of ancestral polymorphism following a recent divergence

Two new Ophiostoma species from Protea caffra in Zambia

The genus Ophiostoma (Ophiostomatales) has a global distribution and species are best known for their association with bark beetles (Curculionidae: Scolytinae) on conifers. An unusual assemblage of these fungi is closely associated with the African endemic plant genus Protea (Proteaceae). Protea-associated Ophiostoma species are ecologically atypical as they colonise the fruiting structures of various serotinous Protea species. Seven species have been described from this niche in South Africa. It has been speculated that novel species may be present in other African countries where these host plants also occur. This view was corroborated by recent collections of two unknown species from Protea caffra trees in Zambia. In the present study we evaluate the species delineation of these isolates using morphological comparisons with other Protea-associated species, differential growth studies and analyses of DNA sequence data for the β-tubulin and internal transcribed spacer (ITS1, 5.8S, ITS2) regions. As a result, the species O. protea-sedis sp. nov., and O. zambiensis sp. nov. are described here as new. This study brings the number of Protea-associated Ophiostoma species to nine and highlights the need for more inclusive surveys, including additional African countries and hosts, to elucidate species diversity in this uncharacteristic niche

Effect of soil type and climatic conditions on the growth and flowering phenology of three Oxalis species in the Western Cape, South Africa

Generally, invasive plant species havewide environmental tolerance that enables them to exploit various climatic conditions and soil types, allowing them to invade new habitats easily. In the Cape Floristic Region, South Africa, the diversity of flowering plants can in part be ascribed to variation in these habitat characteristics that limit their distribution. Climate change has been predicted to bring warmer and drier conditions to the region, with possible influences on the climatic barriers that influence species distributions. We tested the effects of soil type and climate on the growth and flowering of the non-weedy Oxalis tomentosa, and the two weedy species Oxalis purpurea and Oxalis pes-caprae on an altitudinal gradient. The three species, all native to the region, exhibit a range of tolerances to environmental conditions: the first is habitat specific whereas the others are both well known indigenous weeds that have wider tolerance. The results showed that O. purpurea tolerates a variety of conditions well, whereas O. pes-caprae is more restricted by soil type, but would potentially profit from future climatic changes. O. tomentosa, when removed from its native habitat, was stressed under all conditions. These results suggest that habitat-restricted species will be threatened if the predicted level of climate change occurs, while invasive weeds will profit. Studying species responses to different environmental conditions is essential in determining future distributions

Future danger posed by fungi in the Ophiostomatales when encountering new hosts

The Ophiostomatales contain pathogens that threaten forests world-wide. Global trade increases encounters with new hosts, with potential devastating consequences. We assessed the danger posed by the movement of Ophiostomatales between different host trees in South Africa. We tested the pathogenicity of five fungal species from native South African trees, and three from exotic trees, on various native and exotic trees. To evaluate the potential of fungi to move to new hosts, we investigated the strength of their associations with arthropod vectors. Results indicate that many fungal species are pathogens of newly encountered and distantly-related hosts. Encounters of pathogens with new hosts are less likely when host plants are distantly related, and outside the host range of boring beetle vectors, which also reduces the chances of vectoring by phoretic mite associates. However, pathogens associated with numerous mite species and wounds are more likely to encounter new hosts and pose future threats.The authors thank the DST/NRF Centre of Excellence in Tree Health Biotechnology (CHTB) for financial support and the South African National Parks Board (SANPARKS) and Western Cape Nature Conservation Board for issuing the necessary collecting permits.http://www.journals.elsevier.com/fungal-ecology2017-08-31hb2016Microbiology and Plant Patholog

Bark and ambrosia beetles (Curculionidae : Scolytinae), their phoretic mites (Acari) and associated Geosmithia species (Ascomycota : Hypocreales) from Virgilia trees in South Africa

Bark and ambrosia beetles are ecologically and economically important phloeophagous insects that often have complex symbiotic relationships with fungi and mites. These systems are greatly understudied in Africa. In the present study we identified bark and ambrosia beetles, their phoretic mites and their main fungal associates from native Virgilia trees in the Cape Floristic Region (CFR) of South Africa. In addition, we tested the ability of mites to feed on the associated fungi. Four species of scolytine beetles were collected from various Virgilia hosts and from across the CFR. All were consistently associated with various Geosmithia species, fungi known from phloeophagous beetles in many parts of the world, but not yet reported as Scolytinae associates in South Africa. Four beetle species, a single mite species and five Geosmithia species were recovered. The beetles, Hapalogenius fuscipennis, Cryphalini sp. 1 and Scolytoplatypus fasciatus were associated with a single species of Elattoma phoretic mite that commonly carried spores of Geosmithia species. Liparthrum sp. 1 did not carry phoretic mites. Similar to European studies, Geosmithia associates of beetles from Virgilia were constant over extended geographic ranges, and species that share the same host plant individual had similar Geosmithia communities. Phoretic mites were unable to feed on their Geosmithia associates, but were observed to feed on bark-beetle larvae within tunnels. This study forms the first African-centred base for ongoing global studies on the associations between arthropods and Geosmithia species. It strengthens hypotheses that the association between Scolytinae beetles and dry-spored Geosmithia species may be more ubiquitous than commonly recognised.DST⁄NRF Centre of Excellence in Tree Health Biotechnology (CHTB)http://www.elsevier.com/locate/funbiohb201

Death of endemic Virgilia oroboides trees in South Africa caused by Diaporthe virgiliae sp. nov.

Numerous dead and dying individuals of the Western Cape endemic tree Virgilia oroboides (Fabaceae) were recently observed within a South African national botanical garden. Root-rot fungi and fungi symbiotic with bark beetles (Curculionidae; Scolytinae) from diseased trees were assessed for their respective roles in V. oroboides mortality. Disease progression was also monitored over 1 year. Fungi were isolated from surface sterilized bark and root samples from diseased trees and provisionally identified using data from the internal transcribed spacer regions (ITS1, ITS2), including the 5 8S rRNA gene (ITS). Pathogenicity of selected fungi towards V. oroboides was tested under field conditions. The pathogenicity of various bark beetle-associated Geosmithia (Hypocreales: Hypocreomycetidae) spp. from V. oroboides were similarly assessed. The only fungus consistently isolated from lesions on the roots and bark of declining V. oroboides, and never from healthy individuals, represented an undescribed Diaporthe (Diaporthales, Diaporthaceae) species that was characterized using molecular (using data from the ITS marker and part of the b-tubulin gene, TUB), cultural and morphological characters. It is an aggressive pathogen of V. oroboides, newly described here as Diaporthe virgiliae sp. nov. Trees of all ages are susceptible to this pathogen with subsequent bark beetle attack of mature trees only. All Geosmithia spp. from beetles and/or infected trees were nonpathogenic towards V. oroboides. Diaporthe virgiliae caused a severe decline in the health of the monitored V. oroboides population over a period of only 1 year and should be considered as a significant threat to these trees.DST/NRF Centre of Excellence in Tree Health Biotechnology (CHTB).http://onlinelibrary.wiley.com/journal/10.1111/(ISSN)1365-3059hb201

Development of polymorphic microsatellite markers for the genetic characterisation of Knoxdaviesia proteae (Ascomycota: Microascales) using ISSR-PCR and pyrosequencing

Knoxdaviesia proteae is one of the first native ophiostomatoid fungi discovered in South Africa, where it consistently occurs in the infructescences of the iconic Cape Biome plant, Protea repens. Although numerous studies have been undertaken to better understand the ecology of K. proteae, many questions remain to be answered, particularly given its unique niche and association with arthropods for dispersal.We describe the development and distribution of microsatellite markers in K. proteae through Interspersed Simple Sequence Repeat-Polymerase Chain Reaction (ISSR-PCR) enrichment and pyrosequencing. A large proportion of the 31492 sequences obtained from sequencing the enriched genomic DNA were characterised by microsatellites consisting of short tandem repeats and di- and tri-nucleotide motifs. Seventeen percent of these microsatellites contained flanking regions sufficient for primer design. Twenty-three primer pairs were tested, of which 12 amplified and 10 generated polymorphic fragments in K. proteae. Half of these could be transferred to the sister species, K. capensis. The developedmarkers will be used to investigate the reproductive system, genetic diversity and dispersal strategies of K. proteae.National Research Foundation (NRF) and the Department of Science and Technology (DST)/NRF Centre of Excellence in Tree Health Biotechnology (CTHB).http://link.springer.com/journal/11557hb201