Random amplified microsatellites (RAMS) analysis ascertains genetic variation of Alternaria alternata causing black spot disease on Carya illinoinensis in South Africa.

Limited information regarding the occurrence of black spot disease of pecan (Carya illinoinensis), caused by A. alternata, in South Africa is known. The pecan industry is growing rapidly, so it is essential to understand the impact of the fungal pathogen to pecan health. In this study, the genetic variation of 364 A. alternata isolates was investigated by two RAMS primers (CCA5 and CGA5). In total, 6,525 alleles were produced, with a minimum of 3,182 alleles on the CGA5 primer and maximum of 3,343 alleles for CCA5 primer. Further analysis of the primers showed relatively low genetic diversity of A. alternata isolate populations, with mean values; (H = 0.12) and Shannon's information index (I = 0.20). The analysis of molecular variance (AMOVA) revealed significant differences between populations, with 88% of the genetic variation was found within populations (Nm = 3.59, PhiPT = 0.12), and were not significantly different (p > 0.001). While 12% variation was observed among populations (Nm = 2.89, PhiPT = 0.08) and the estimates were statistically significant (p < 0.001). STRUCTURE HARVESTER output showed that K value is K = 8, where ΔK cannot find the true number of populations because of less variation. The dendrogram cluster tree generated by Ward's analysis unveiled two main distinct clades and 10 sub-clades, revealing similar findings as those of PCoA analysis clusters. Therefore, it was evident that these analyses depicted no distinct relationship between the A. alternata isolates and their geographic locations or the prevalence of distribution among the populations.

In Vitro Evaluation of Azoxystrobin, Boscalid, Fentin-Hydroxide, Propiconazole, Pyraclostrobin Fungicides against Alternaria alternata Pathogen Isolated from Carya illinoinensis in South Africa.

Black spot disease or Alternaria black spot (ABS) of pecan (Carya illinoinensis) in South Africa is caused by Alternaria alternata. This fungal pathogen impedes the development of pecan trees and leads to low yield in pecan nut production. The present study investigated the in vitro effect of six fungicides against the mycelial growth of A. alternata isolates from ABS symptoms. Fungicides tested include Tilt (propiconazole), Ortiva (azoxystrobin), AgTin (fentin hydroxide), and Bellis (boscalid + pyraclostrobin). All fungicides were applied in 3 concentrations (0.2, 1, and 5 µg mL-1). Tilt and Bumper 250 EC containing propiconazole active ingredient (demethylation Inhibitors) were the most effective and inhibited all mycelial growth from up to 6 days post-incubation. The other active ingredients (succinate dehydrogenase inhibitors, organotin compounds, and quinone outside inhibitors) showed 75-85% mycelial growth inhibition. The effective concentration to inhibit mycelial growth by 50% (EC50) was estimated for each isolate and fungicide. The overall mean EC50 values for each fungicide on the six isolates were 1.90 µg mL-1 (Tilt), 1.86 µg mL-1 (Ortiva), 1.53 µg mL-1 (AgTin), and 1.57 µg mL-1 for (Bellis). This initial screening suggested that propiconazole fungicide was the most effective for future field trials test and how these fungicides could be used in controlling ABS disease.

Global species diversity and distribution of the psychedelic fungal genus Panaeolus.

Psychedelic fungi have received considerable attention recently due to their promising treatment potential of several psychiatric disorders and medical conditions, both in clinical settings but also as a nutraceutical. Besides research, a growing number of companies are developing capacity to conduct research and clinical trials where these fungi and their products can be used, and to provide these fungi to the public market that are rapidly becoming legal across the world. Whereas Psilocybe species are better known as psychedelic fungi, species in Panaeolus are also reputed to contain the psychedelic compound psilocybin and used recreationally. For the novice, there is no contemporary scientific summary of all the species in this genus that are known to be psychedelic, compared to those that are not. The global distribution and species diversity of these brown to white, often inconspicuous mushrooms are also not summarised, nor is it known to what extent DNA sequence data that are needed for identification have been generated for all of the species in this genus. However, psychedelic Panaeolus species are used and moved across the world. This lack of data makes it difficult to regulate bioexploitation and apply law enforcement of these fungi and the compounds they contain, especially seen in the light of the rapid development of the related markets. The aim of this review is to summarise current scientific data and knowledge on the species biodiversity, geographical distribution, extent of sequence data for identification purposes, and the psychedelic potential of species, based on published results. The review revealed where species are mostly known from, while also indicating areas seriously lacking such biodiversity data. A significant degree of study across the world is still needed to confirm which of these species are truly psychedelic and exactly what compounds they can produce.

Differential Detection of Alternaria alternata Haplotypes Isolated from Carya illinoinensis Using PCR-RFLP Analysis of Alt a1 Gene Region.

Alternaria black spot disease on pecan is caused by the opportunistic pathogen Alternaria alternata and poses a serious threat to the local South African and global pecan industry. Several diagnostic molecular marker applications have been established and used in the screening of various fungal diseases worldwide. The present study investigated the potential for polymorphism within samples of A. alternata isolates obtained from eight different geographical locations in South Africa. Pecan (Carya illinoinensis) leaves, shoots, and nuts-in-shuck with Alternaria black spot disease were sampled, and 222 A. alternata isolates were retrieved. For rapid screening to identify Alternaria black spot pathogens, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the Alternaria major allergen (Alt a1) gene region was used, followed by the digestion of the amplicons with HaeIII and HinfI endonucleases. The assay resulted in five (HaeIII) and two (HinfI) band patterns. Unique banding patterns from the two endonucleases showed the best profile and isolates were grouped into six clusters using a UPGMA (unweighted pair group method with arithmetic averages) distance matrix (Euclidean) dendrogram method on R-Studio. The analysis confirmed that the genetic diversity of A. alternata does not depend on host tissues or the pecan cultivation region. The grouping of selected isolates was confirmed by DNA sequence analysis. The Alt a1 phylogeny corroborated no speciation within the dendrogram groups and showed 98-100% bootstrap similarity. This study reports the first documented rapid and reliable technique for routine screening identification of pathogens causing Alternaria black spot in South Africa.

Multigene Phylogeny and Pathogenicity Trials Revealed Alternaria alternata as the Causal Agent of Black Spot Disease and Seedling Wilt of Pecan (Carya illinoinensis) in South Africa.

The pecan (Carya illinoinensis) industry in South Africa is growing rapidly, and it is becoming increasingly crucial to understand the risks posed to pecans by fungal pathogens. Black spots on leaves, shoots, and nuts in shucks caused by Alternaria species have been observed since 2014 in the Hartswater region of the Northern Cape Province of South Africa. Species of Alternaria include some of the most ubiquitous plant pathogens on earth. The aim of this study was to use molecular techniques to identify the causative agents of Alternaria black spot and seedling wilt isolated from major South African pecan-production areas. Symptomatic and non-symptomatic pecan plant organs (leaves, shoots, and nuts-in-shucks) were collected from pecan orchards, representing the six major production regions in South Africa. Thirty Alternaria isolates were retrieved from the sampled tissues using Potato Dextrose Agar (PDA) culture media and molecular identification was conducted. The phylogeny of multi-locus DNA sequences of Gapdh, Rpb2, Tef1, and Alt a 1 genes revealed that the isolates were all members of Alternaria alternata sensu stricto, forming part of the Alternaria alternata species complex. The virulence of six A. alternata isolates were tested on detached nuts of Wichita and Ukulinga cultivars, respectively, as well as detached leaves of Wichita. The A. alternata isolates were also evaluated for their ability to cause seedling wilt in Wichita. The results differed significantly between wounded and unwounded nuts of both cultivars, but not between the cultivars. Similarly, the disease lesions on the wounded detached leaves were significantly different in size from the unwounded leaves. The seedling tests confirmed that A. alternata is pathogenic and that A. alternata causes black spot disease and seedling wilt of pecans. This study is one of the first documentations of Alternaria black spot disease of pecan trees and its widespread occurrence in South Africa.

Filamentous fungi for sustainable remediation of pharmaceutical compounds, heavy metal and oil hydrocarbons.

This review presents a comprehensive summary of the latest research in the field of bioremediation with filamentous fungi. The main focus is on the issue of recent progress in remediation of pharmaceutical compounds, heavy metal treatment and oil hydrocarbons mycoremediation that are usually insufficiently represented in other reviews. It encompasses a variety of cellular mechanisms involved in bioremediation used by filamentous fungi, including bio-adsorption, bio-surfactant production, bio-mineralization, bio-precipitation, as well as extracellular and intracellular enzymatic processes. Processes for wastewater treatment accomplished through physical, biological, and chemical processes are briefly described. The species diversity of filamentous fungi used in pollutant removal, including widely studied species of Aspergillus, Penicillium, Fusarium, Verticillium, Phanerochaete and other species of Basidiomycota and Zygomycota are summarized. The removal efficiency of filamentous fungi and time of elimination of a wide variety of pollutant compounds and their easy handling make them excellent tools for the bioremediation of emerging contaminants. Various types of beneficial byproducts made by filamentous fungi, such as raw material for feed and food production, chitosan, ethanol, lignocellulolytic enzymes, organic acids, as well as nanoparticles, are discussed. Finally, challenges faced, future prospects, and how innovative technologies can be used to further exploit and enhance the abilities of fungi in wastewater remediation, are mentioned.

Psilocybin containing mushrooms: a rapidly developing biotechnology industry in the psychiatry, biomedical and nutraceutical fields.

Humans have collected and used hallucinogenic mushrooms for ethnic medicinal, recreational, and religious purposes since before recorded history. Currently, the use of these mushrooms is illegal in most countries, but where their use is legal they are applied as self medication. Psilocybin and psilocin, two psychoactive alkaloids, are naturally synthesized by hallucinogenic mushrooms. The chemical structure of these compounds are similar to the neurotransmitter serotonin. Activation of this system by psilocybin and psilocin may produce temporary changes in the brain that induce hallucinations and feelings of euphoria. Adjustment of the serotonin system in this way can moderate symptoms of related mental disorders. This review summarizes relevant and current information regarding the discovery of hallucinogenic mushrooms and their contained psychoactive compounds, the events that lead to their criminalization and decriminilization, and the state of knowledge of psilocybin, psilocin, and derivatives. Last, research on the psychoactive properties of these mushrooms is placed in perspective to possible applications for human dysfunctions.

Plant and Soil Core Mycobiomes in a Two-Year Sorghum-Legume Intercropping System of Underutilized Crops in South Africa.

Fungal communities form close beneficial (mutualists) or detrimental (pathogens) associations with their plant hosts. Their diversity and abundance can be affected by agricultural practices which include cropping systems such as rotations and intercropping. Despite the importance of cropping systems in increasing productivity, knowledge of the fungal mycobiome and the core inhabitants for under-utilised cereal and legume crops, particularly over a period, is still limited. The core mycobiomes in plant tissues and bulk soils of a cereal-legume intercrop were characterized over two years using high-throughput sequencing. The intercropping trial consisted of sorghum, Bambara groundnut, cowpea, dry bean, and soybean. A greater number of molecular operational taxonomic units (MOTUs) were found in plant tissues compared to those from the soils and between year one and year two. Principal coordinate analyses revealed that fungal communities for each year were relatively distinct, particularly for the soils. The core mycobiome was dominated by a Davidiellaceae sp. (Cladosporium), Didymellaceae sp. 1 (Phoma), Didymellaceae sp. 2 (Epicoccum), Fusarium sp. 2, Unidentified (Ascomycota), and Cryptococcus MOTUs that were present in all plant tissues and soils of year one and two. Other key MOTUs were only specific to a year, substrate, or crop. Although the mycobiome of sorghum were more distinct than the cores of the legumes, there were still MOTUs dominant across all of the crops. Characterization of this baseline core across two years provides insight into those fungi that are always present in these crops, and that could be utilized in improving crop performance and productivity.

Baseline Data of the Fungal Phytobiome of Three Sorghum (Sorghum bicolor) Cultivars in South Africa using Targeted Environmental Sequencing.

Plant-associated fungi, or the mycobiome, inhabit plant surfaces above ground, reside in plant tissues as endophytes, or are rhizosphere in the narrow zone of soil surrounding plant roots. Studies have characterized mycobiomes of various plant species, but little is known about the sorghum mycobiome, especially in Africa, despite sorghum being one of the most important indigenous and commercial cereals in Africa. In this study, the mycobiome associated with above- and below-ground tissues of three commercial sorghum cultivars, as well as from rhizosphere and surrounding bulk soil samples, were sequenced using targeted sequencing with the Illumina MiSeq platform. Relative abundance differences between fungal communities were found between above-ground and below-ground niches, with most differences mostly in the dominant MOTUs, such as Davidiellaceae sp. (Cladosporium), Didymellaceae sp. 1 (Phoma), Fusarium, Cryptococcus and Mucor. Above-ground communities also appeared to be more diverse than below-ground communities, and plants harboured the most diversity. A considerable number of MOTUs were shared between the cultivars although, especially for NS5511, their abundances often differed. Several of the detected fungal groups include species that are plant pathogens of sorghum, such as Fusarium, and, at low levels, Alternaria and the Ustilaginomycetes. Findings from this study illustrate the usefulness of targeted sequencing of the ITS rDNA gene region (ITS2) to survey and monitor sorghum fungal communities and those from associated soils. This knowledge may provide tools for disease management and crop production and improvement.

One-pot synthesis of zinc oxide nanoparticles via chemical precipitation for bromophenol blue adsorption and the antifungal activity against filamentous fungi.

In this research, zinc oxide nanoparticles (ZnONPs) were prepared via a facile one-pot chemical precipitation approach and applied in the adsorption of bromophenol blue (BRB) and as antifungal agents against the filamentous fungi and plant pathogens; Alternaria alternata CGJM3078, Alternaria alternata CGJM3006 and Fusarium verticilliodes CGJM3823. The ZnONPs were characterized by the UV-Vis, FTIR, XRD, TGA, BET, SEM, TEM, and EDX techniques, which showed efficient synthesis. The characteristics ZnO UV-Vis absorption band was observed at 375 nm, while the XRD showed an average ZnONPs crystalline size of 47.2 nm. The SEM and TEM images showed an irregular shaped and aggregated porous structure of 65.3 nm average-sized ZnONPs. The TGA showed 22.9% weight loss at 800 °C indicating the high thermal stability of ZnONPs, while BET analysis revealed a surface area, pore volume and pore diameter of 9.259 m2/g, 0.03745 cm3/g and 9.87 nm respectively. The Freundlich, pseudo-second-order, and intra-particle diffusion models showed R2 > 0.9494 and SSE < 0.7412, thus, exhibited the best fit to the isotherm and kinetics models. Thermodynamics revealed feasible, endothermic, random, and spontaneous adsorption of BRB onto the synthesized ZnONPs. The antifungal assay conducted depicts strong antifungal activities against all three tested fungi. Noticeably, ZnONPs (0.002-5 mg/mL) showed maximum activities with the largest zone of inhibition against A. alternata CGJM 3006 from 25.09 to 36.28 mm. This was followed by the strain F. verticilliodes CGJM 3823 (range from 23.77 to 34.77 mm) > A. alternata CGJM3078 (range from 22.73 to 30.63 mm) in comparison to Bleach 5% (positive control). Additionally a model was proposed based on the possible underlying mechanisms for the antifungal effect. This research demonstrated the potent use of ZnONPs for the adsorption of BRB and as effective antifungal agents.

High genetic diversity of spider species in a mosaic montane grassland landscape.

Gene flow and genetic variation were examined within and among populations of five of the most common spider species in shrublands of the mountainous Golden Gate Highlands National Park (GGHNP), South Africa. These species included three active hunters, Dendryphantes purcelli Peckham & Peckham, 1903 (Salticidae), Pherecydes tuberculatus O.P.-Cambridge, 1883 (Thomisidae) and Philodromus browningi Lawrence, 1952 (Philodromidae), and two web-builders, Neoscona subfusca (C.L. Koch, 1837) (Araneidae) and a Theridion Walckenaer, 1802 species (Theridiidae). A total of 249 spiders (57 D. purcelli, 69 N. subfusca, 34 P. browningi, 56 P. tuberculatus and 33 Theridion sp.) were collected and analysed from six shrubland localities in the park. Analyses of sequence variation of the mitochondrial cytochrome oxidase c subunit I (COI) gene for each species revealed relatively low nucleotide diversity (π < 0.0420) but high genetic diversity (Hd > 0.6500) within populations for all species, except P. tuberculatus. Genetic differentiation was also noted to differ between species, with only P. tuberculatus indicating very large divergence (Fst > 0.2500). These results were reflected by gene flow, with D. purcelli, N. subfusca and the Theridion sp. estimated as experiencing more than one disperser per generation. Overall, highest gene flow was found in the two web-building species, indicating possible high dispersal ability of these spiders in the GGHNP. Additionally, constructed phylogenies indicated possible cryptic speciation occurring in the majority of the investigated species. Our current results indicate that the five investigated spider species were able to maintain gene flow between shrubland populations within the GGHNP to some degree, despite the mountainous landscape. However, further analyses incorporating additional molecular markers are needed to properly determine the extent of genetic diversity and gene flow of these species within the GGHNP.

A first checklist of macrofungi for South Africa.

Macrofungi are considered as organisms that form large fruiting bodies above or below the ground that are visible without the aid of a microscope. These fungi include most basidiomycetes and a small number of ascomycetes. Macrofungi have different ecological roles and uses, where some are edible, medicinal, poisonous, decomposers, saprotrophs, predators and pathogens, and they are often used for innovative biotechnological, medicinal and ecological applications. However, comprehensive checklists, and compilations on the diversity and distribution of mushrooms are lacking for South Africa, which makes regulation, conservation and inclusion in national biodiversity initiatives difficult. In this review, we compiled a checklist of macrofungi for the first time (excluding lichens). Data were compiled based on available literature in journals, books and fungorium records from the National Collection of Fungi. Even if the list is not complete due to numerous unreported species present in South Africa, it still represents an overview of the current knowledge of the macromycetes of South Africa. The list of names enables the assessment of gaps in collections and knowledge on the fungal biodiversity of South Africa, and downstream applications such as defining residency status of species. It provides a foundation for new names to be added in future towards developing a list that will be as complete as possible, and that can be used by a wide audience including scientists, authorities and the public.

Bioactive compounds from the endophytic fungus Fusarium proliferatum.

The crude extract of an endophytic fungus isolated from Syzygium cordatum and identified as Fusarium proliferatum showed 100% cytotoxicity against the brine shrimp Artemia salina at 100 µg/mL. Seven coloured, biologically active metabolites - including ergosta-5,7,22-trien-3ß-ol, nectriafurone-8-methyl ether, 9-O-methyl fusarubin, bostrycoidin, bostrycoidin-9-methyl ether and 8-hydroxy-5,6-dimethoxy-2-methyl-3-(2-oxo-propyl)-1,4-naphthoquinone- were isolated from the extract.

Multiple Phytophthora species associated with a single riparian ecosystem in South Africa.

The diversity of Phytophthora spp. in rivers and riparian ecosystems has received considerable international attention, although little such research has been conducted in South Africa. This study determined the diversity of Phytophthora spp. within a single river in Gauteng province of South Africa. Samples were collected over 1 y including biweekly river baiting with Rhododendron indicum leaves. Phytophthora isolates were identified with phylogenetic analyses of sequences for the internal transcribed spacer (ITS) region of the ribosomal DNA and the mitochondrial cytochrome oxidase c subunit I (coxI) gene. Eight Phytophthora spp. were identified, including a new taxon, P. taxon Sisulu-river, and two hybrid species from Cooke's ITS clade 6. Of these, species from Clade 6 were the most abundant, including P. chlamydospora and P. lacustris. Species residing in Clade 2 also were encountered, including P. multivora, P. plurivora and P. citrophthora. The detection of eight species in this investigation of Phytophthora diversity in a single riparian river ecosystem in northern South Africa adds to the known diversity of this genus in South Africa and globally.

Recommendations of generic names in Diaporthales competing for protection or use.

In advancing to one name for fungi, this paper treats generic names competing for use in the order Diaporthales (Ascomycota, Sordariomycetes) and makes a recommendation for the use or protection of one generic name among synonymous names that may be either sexually or asexually typified. A table is presented that summarizes these recommendations. Among the genera most commonly encountered in this order, Cytospora is recommended over Valsa and Diaporthe over Phomopsis. New combinations are introduced for the oldest epithet of important species in the recommended genus. These include Amphiporthe tiliae, Coryneum lanciforme, Cytospora brevispora, C. ceratosperma, C. cinereostroma, C. eugeniae, C. fallax, C. myrtagena, Diaporthe amaranthophila, D. annonacearum, D. bougainvilleicola, D. caricae-papayae, D. cocoina, D. cucurbitae, D. juniperivora, D. leptostromiformis, D. pterophila, D. theae, D. vitimegaspora, Mastigosporella georgiana, Pilidiella angustispora, P. calamicola, P. pseudogranati, P. stromatica, and P. terminaliae.

Finding needles in haystacks: linking scientific names, reference specimens and molecular data for Fungi.

DNA phylogenetic comparisons have shown that morphology-based species recognition often underestimates fungal diversity. Therefore, the need for accurate DNA sequence data, tied to both correct taxonomic names and clearly annotated specimen data, has never been greater. Furthermore, the growing number of molecular ecology and microbiome projects using high-throughput sequencing require fast and effective methods for en masse species assignments. In this article, we focus on selecting and re-annotating a set of marker reference sequences that represent each currently accepted order of Fungi. The particular focus is on sequences from the internal transcribed spacer region in the nuclear ribosomal cistron, derived from type specimens and/or ex-type cultures. Re-annotated and verified sequences were deposited in a curated public database at the National Center for Biotechnology Information (NCBI), namely the RefSeq Targeted Loci (RTL) database, and will be visible during routine sequence similarity searches with NR_prefixed accession numbers. A set of standards and protocols is proposed to improve the data quality of new sequences, and we suggest how type and other reference sequences can be used to improve identification of Fungi. Database URL: http://www.ncbi.nlm.nih.gov/bioproject/PRJNA177353.

Botryosphaeriaceae species overlap on four unrelated, native South African hosts.

Botryosphaeriaceae represents an important and diverse family of latent fungal pathogens of woody plants. We address the question of host range of these fungi by sampling leaves and branches of four native South African trees, including Acacia karroo (Fabaceae), Celtis africana (Cannabaceae), Searsia lancea (Anacardiaceae), and Gymnosporia buxifolia (Celastraceae). Two new species of the Botryosphaeriaceae, namely Tiarosporella africana sp. nov. and Aplosporella javeedii sp. nov. were identified, together with five known species, including Neofusicoccum parvum, Neofusicoccum kwambonambiense, Spencermartinsia viticola, Diplodia pseudoseriata, and Botryosphaeria dothidea. Most Botryosphaeriaceae occurred on more than one host. With the exception of S. lancea, which was infected by A. javeedii all the hosts were infected by more than one Botryosphaeriaceae species. Collectively, the results suggest that some intrinsic host factors, possibly combined with local environmental conditions, affect the distribution and co-infectivity of various hosts by the Botryosphaeriaceae. This would counteract the general ability of a species in the Botryosphaeriaceae to infect a broad range of plants. The combination of host and environmental factors might also explain why some Botryosphaeriaceae with apparently broad host ranges, are found on different suites of hosts in different areas of the world.

Ion Torrent PGM as tool for fungal community analysis: a case study of endophytes in Eucalyptus grandis reveals high taxonomic diversity.

The Kingdom Fungi adds substantially to the diversity of life, but due to their cryptic morphology and lifestyle, tremendous diversity, paucity of formally described specimens, and the difficulty in isolating environmental strains into culture, fungal communities are difficult to characterize. This is especially true for endophytic communities of fungi living in healthy plant tissue. The developments in next generation sequencing technologies are, however, starting to reveal the true extent of fungal diversity. One of the promising new technologies, namely semiconductor sequencing, has thus far not been used in fungal diversity assessments. In this study we sequenced the internal transcribed spacer 1 (ITS1) nuclear encoded ribosomal RNA of the endophytic community of the economically important tree, Eucalyptus grandis, from South Africa using the Ion Torrent Personal Genome Machine (PGM). We determined the impact of various analysis parameters on the interpretation of the results, namely different sequence quality parameter settings, different sequence similarity cutoffs for clustering and filtering of databases for removal of sequences with incomplete taxonomy. Sequence similarity cutoff values only had a marginal effect on the identified family numbers, whereas different sequence quality filters had a large effect (89 vs. 48 families between least and most stringent filters). Database filtering had a small, but statistically significant, effect on the assignment of sequences to reference sequences. The community was dominated by Ascomycota, and particularly by families in the Dothidiomycetes that harbor well-known plant pathogens. The study demonstrates that semiconductor sequencing is an ideal strategy for environmental sequencing of fungal communities. It also highlights some potential pitfalls in subsequent data analyses when using a technology with relatively short read lengths.

Surveys of soil and water reveal a goldmine of Phytophthora diversity in South African natural ecosystems.

Phytophthora species are well-known as destructive plant pathogens, especially in natural ecosystems. It is ironic, therefore, how little is known regarding the Phytophthora diversity in South African natural woody ecosystems. In this study, Phytophthora species were isolated using standard baiting techniques from 182 soil and water samples and these were identified based on ITS and coxI sequence data. The 171 resulting Phytophthora isolates resided in 14 taxa including six known species (P. multivora, P. capensis, P. cryptogea, P. frigida, P. cinnamomi, P. cinnamomi var. parvispora), the known but as yet unnamed Phytophthora sp. PgChlamydo, P. sp. emzansi, and P. sp. Kununurra and five novel taxa referred to as P. sp. stellaris, P. sp. Umtamvuna P. sp. canthium, P. sp. xWS, P. sp. xHennops. Four of the new taxa were found exclusively in water and two of these are hybrids. The most commonly isolated species from soil was P. multivora, a species recently described from Western Australia. Phytophthora frigida was isolated for the first time from stream water. With the exception of P. cinnamomi, very little is known regarding the biology, epidemiology or origin of Phytophthora in South Africa.

Characterization of Phytophthora hybrids from ITS clade 6 associated with riparian ecosystems in South Africa and Australia.

Surveys of Australian and South African rivers revealed numerous Phytophthora isolates residing in clade 6 of the genus, with internal transcribed spacer (ITS) gene regions that were either highly polymorphic or unsequenceable. These isolates were suspected to be hybrids. Three nuclear loci, the ITS region, two single copy loci (antisilencing factor (ASF) and G protein alpha subunit (GPA)), and one mitochondrial locus (cytochrome oxidase c subunit I (coxI)) were amplified and sequenced to test this hypothesis. Abundant recombination within the ITS region was observed. This, combined with phylogenetic comparisons of the other three loci, confirmed the presence of four different hybrid types involving the three described parent species Phytophthora amnicola, Phytophthora thermophila, and Phytophthora taxon PgChlamydo. In all cases, only a single coxI allele was detected, suggesting that hybrids arose from sexual recombination. All the hybrid isolates were sterile in culture and all their physiological traits tended to resemble those of the maternal parents. Nothing is known regarding their host range or pathogenicity. Nonetheless, as several isolates from Western Australia were obtained from the rhizosphere soil of dying plants, they should be regarded as potential threats to plant health. The frequent occurrence of the hybrids and their parent species in Australia strongly suggests an Australian origin and a subsequent introduction into South Africa.