Longevity, population stage and size structures, morphology and reproduction of four long-lived grassland suffrutices

MSc., Faculty of Science, University of the Witwatersrand, 2011Little is known about the longevity of grassland suffrutex plants and the relationship between longevity and plant morphological structures and seedling growth strategies. The aim of this study was to determine the longevity of four grassland suffrutex plant species, namely Berkheya insignis, Callilepis laureola, Protea insignis and Tephrosia kraussiana. Seed viability, seedling growth, morphology, habitat and population structure and demography were also assessed and related to plant age. The rhizomes, seeds (if available) and canopies of B. insignis, C. laureola, P. simplex and T. kraussiana were removed from a site near Port Edward in the Pondoland region, KwaZulu-Natal, South Africa in April 2008. Seeds were available for P. simplex and T. kraussiana only. Seed germination and viability were tested in the field and laboratory using germination trials and tetrazolium tests. Greenhouse and field grown seedlings were used to monitor seedling growth and to record seedling morphology. The aerial and rhizome morphologies of adult plants excavated from the field were also recorded. The largest of these rhizomes were aged using radiocarbon dating. Attempts were made to develop morphological surrogates for plant age as no method currently exists. Plant density, demographies, number of inflorescences and various environmental variables of wild populations of B. insignis, C. laureola, P. simplex and T. kraussiana were sampled from the Red Desert Nature Reserve and the Umtamvuna Nature Reserve. Species morphologies varied, however important similarities suggest convergent evolution. These included a single vertical main stem rhizome with multiple side branches, early rhizome development in seedlings, and high root: shoot ratios. Seed viability was high in the laboratory but low seedling emergence was observed in the field. Seedling growth was rapid for T. kraussiana and slow for P. simplex. The population stage structure for all four species comprised primarily adults with few or no juveniles. Wild population plant height and main stem diameter followed normal distributions. Number of stems, basal area, canopy area and the number of floral structures had distributions favouring the smaller size classes. Soil P, N, K and organic carbon were important soil nutrients in a PCA analysis of the habitats of the four species. Radiocarbon dating yielded the following ages: B. insignis: 49-51 years, C. laureola: 49-50 years, P. simplex: 49-51 years and T. kraussiana: 51 years. There was a significant relationship between rhizome mass and canopy area, basal area, height, number of stems and main stem diameter. Since rhizome mass had a positive relationship with age- a relationship between age and aerial structures is likely. Therefore, creating surrogates for age may be possible. Overall, these species have moderate longevity, are poor seed producers with possibly slow population growth and are closely associated with soil nutrients. Therefore, these and other suffrutex species are particularly vulnerable to habitat destruction and climate change. The results of this study indicate that there needs to be a greater focus on below ground growth during ecological assessments in order to better understand the ecology of our diverse grassland biome plants

Long term stability and infectivity of herpesviruses in water

For viruses to utilize environmental vectors (hard surfaces, soil, water) for transmission, physical and chemical stability is a prerequisite. There are many factors including pH, salinity, temperature, and turbidity that are known to contribute to the ability of viruses to persist in water. Equine herpesvirus type-1 (EHV-1) is a pathogenic alphaherpesvirus associated with domestic horses and wild equids. EHV-1 and recombinants of EHV-1 and EHV-9 are able to cause infections in non-equid animal species, particularly in captive settings. Many of the captive non-equid mammals are not naturally sympatric with equids and do not share enclosures, however, in many cases water sources may overlap. Similarly, in the wild, equids encounter many species at waterholes in times of seasonal drought. Therefore, we hypothesized that EHV-1 is stable in water and that water may act as a vector for EHV-1. In order to establish the conditions promoting or hindering EHV-1 longevity, infectivity and genomic stability in water; we exposed EHV-1 to varied water environments (pH, salinity, temperature, and turbidity) in controlled experiments over 21 days. The presence and infectivity of the virus was confirmed by both qPCR and cell culture experiments. Our results show that EHV-1 remains stable and infectious under many conditions in water for up to three weeks

Degradation and remobilization of endogenous retroviruses by recombination during the earliest stages of a germ-line invasion

Endogenous retroviruses (ERVs) are proviral sequences that result from colonization of the host germ line by exogenous retroviruses. The majority of ERVs represent defective retroviral copies. However, for most ERVs, endogenization occurred millions of years ago, obscuring the stages by which ERVs become defective and the changes in both virus and host important to the process. The koala retrovirus, KoRV, only recently began invading the germ line of the koala (Phascolarctos cinereus), permitting analysis of retroviral endogenization on a prospective basis. Here, we report that recombination with host genomic elements disrupts retroviruses during the earliest stages of germ-line invasion. One type of recombinant, designated recKoRV1, was formed by recombination of KoRV with an older degraded retroelement. Many genomic copies of recKoRV1 were detected across koalas. The prevalence of recKoRV1 was higher in northern than in southern Australian koalas, as is the case for KoRV, with differences in recKoRV1 prevalence, but not KoRV prevalence, between inland and coastal New South Wales. At least 15 additional different recombination events between KoRV and the older endogenous retroelement generated distinct recKoRVs with different geographic distributions. All of the identified recombinant viruses appear to have arisen independently and have highly disrupted ORFs, which suggests that recombination with existing degraded endogenous retroelements may be a means by which replication-competent ERVs that enter the germ line are degraded

Seasonal host and ecological drivers may promote restricted water as a viral vector

In climates with seasonally limited precipitation, terrestrial animals congregate at high densities at scarce water sources. We hypothesize that viruses can exploit the recurrence of these diverse animal congrega- tions to spread. In this study, we test the central prediction of this hypothesis — that viruses employing this transmission strategy remain stable and infectious in water. Equid herpesviruses (EHVs) were cho- sen as a model as they have been shown to remain stable and infectious in water for weeks under labo- ratory conditions. Using fecal data from wild equids from a previous study, we establish that EHVs are shed more frequently by their hosts during the dry season, increasing the probability of water source contamination with EHV. We document the presence of several strains of EHVs present in high genome copy number from the surface water and sediments of waterholes sampled across a variety of mamma- lian assemblages, locations, temperatures and pH. Phylogenetic analysis reveals that the different EHV strains found exhibit little divergence despite representing ancient lineages. We employed molecular approaches to show that EHVs shed remain stable in waterholes with detection decreasing with increas- ing temperature in sediments. Infectivity experiments using cell culture reveals that EHVs remain infectious in water derived from waterholes. The results are supportive of water as an abiotic viral vector for EHVacceptedVersio

Retroviral integrations contribute to elevated host cancer rates during germline invasion

© 2021, The Author(s). Repeated retroviral infections of vertebrate germlines have made endogenous retroviruses ubiquitous features of mammalian genomes. However, millions of years of evolution obscure many of the immediate repercussions of retroviral endogenisation on host health. Here we examine retroviral endogenisation during its earliest stages in the koala (Phascolarctos cinereus), a species undergoing germline invasion by koala retrovirus (KoRV) and affected by highcancerprevalence. We characterise KoRV integration sites (IS) in tumour and healthy tissues from 10 koalas, detecting 1002 unique IS, with hotspots of integration occurring in the vicinity of known cancer genes. We find that tumours accumulate novel IS, with proximate genes over-represented for cancer associations. We detect dysregulation of genes containing IS and identify a highly-expressed transduced oncogene. Our data provide insights into the tremendous mutational load suffered by the host during active retroviral germline invasion, a process repeatedly experienced and overcome during the evolution of vertebrate lineages

Discovery of novel circular replication-associated protein encoding single-stranded DNA viruses in ecosystems using viral metagenomic approaches

The introduction of next-generation sequencing (NGS) technologies has dramatically changed the field of virology, with many significant discoveries of novel circular replication-associated protein (Rep) encoding single-stranded (CRESS) DNA viruses. Traditionally, most research into CRESS DNA viruses has often focused on investigating plant and animal pathogens that are of significant economic importance. This research has led to the discovery and establishment of three different CRESS DNA families including Geminiviridae, Nanoviridae and Circoviridae, which infect eukaryotes. CRESS DNA viruses can have single or multicomponent genomes, with the latter requiring all components for infection. CRESS DNA viruses have circular single-stranded DNA (ssDNA) genomes with at least one protein encoding a Rep which is responsible for viral replication. It has been shown that CRESS DNA viruses are able to evolve rapidly with nucleotide substitution rates that are similar to those observed in RNA viruses. The Rep gene has conserved regions known as motifs which are often used to determine relatedness between CRESS DNA virus. NGS has expanded our knowledge on the diversity of novel CRESS DNA viruses. Viral genomes are now routinely recovered from different sample types without any prior knowledge of the viral sequence. This has led to the development of the field of viral ecology. This field places an emphasis on viruses being one of the most abundant organisms on earth, and are therefore likely to play a major role in ecosystems. Environmental metagenomic studies have isolated CRESS DNA viruses from sea water, freshwater, faecal matter from various animals, soil, the atmosphere, sediments and sewage; dramatically increasing the known CRESS DNA viral genomes in the public domain. These studies are shedding light on the distribution of CRESS DNA viruses, as well as providing baseline data for future studies to examine virus-host interactions, community structure and ultimately viral evolution. Vector enable metagenomics (VEM) is another novel approach utilising NGS techniques for discovering CRESS DNA viruses. As many plant-infecting CRESS DNA viruses such as geminiviruses and nanoviruses are vectored by insects, this approach exploits this mechanism by using insect vectors as a surveillance tool to monitor and survey these viruses circulating in ecosystems. Recent studies have used these methods to identify known viral plant pathogens as well as novel viruses circulating in insect vectors such as whiteflies and other higher order insects such a mosquitoes and dragonflies. These approaches successfully demonstrated that VEM can be used as a unique method, with the first mastrevirus discovered in the new world being recovered from dragonfly species Erythrodiplax fusca using this approach. The research in this thesis uses metagenomics to survey CRESS DNA viral diversity in different organisms and environments. Two hundred and sixty eight novel CRESS DNA viruses were recovered and verified in this study from a range of sample types (adult Odonata, Odonata larvae, Mollusca, benthic sediment, water, Oligochaeta and Chironomidae) collected in the United States of America, Australia and New Zealand. All viral genomes isolated had two major proteins encoding for a putative Rep and coat protein (CP), with major Rep motifs identified in most Reps. Phylogenetic analysis of the Reps encoded by the viral genomes highlighted that most were extremely diverse falling outside of the previously described ssDNA viral families. A top-down approach was implemented to recover CRESS DNA viruses and possible viral pathogens from Odonata and their larvae. Thirty six viral genomes were recovered from terrestrial adult dragonflies as well as the twenty four from aquatic larvae. Dragonfly cycloviruses were isolated from the some adult Odonata species which were closely related to the isolates previously described by Rosario et al. (2012). The viruses isolated in the aquatic and terrestrial ecosystems differed substantially indicating that different CRESS DNA viromes exist in both land and water. The diversity of CRESS DNA viruses in seven different mollusc species (Amphibola crenata, Austrolvenus stutchburyi, Paphies subtriangulata, Musculium novazelandiae, Potamopyrgus antipodarum, Physella acuta and Echyridella menziesi) from Lake Sarah and the Avon-Heathcote estuary both in New Zealand, were also investigated. One hundred and forty nine novel viral genomes were recovered. Two CRESS DNA genomes were recovered from molluscs which have Rep-like sequences most closely related to those found in some bacterial genomes. Sclerotinia sclerotiorum hypovirulence-associated DNA virus 1 (SsHADV-1) was originally isolated from fungal species Sclerotinia sclerotiorum in china and was later found in benthic sediments in New Zealand. As part of this study, SsHADV-1 was recovered from dragonflies (Erythemis simplicicollis, Ischnura ramburii and Pantala hymenaea) collected in Arizona and Oklahoma, USA suggesting a larger distribution of these viruses and not surprising given the near global distribution of S. sclerotiorum. Dragonfly larvae-associated circular DNA viruses (DflaCVs) that were originally isolated in Odonata larvae samples from three New Zealand lakes were later recovered from water, benthic sediment, worms and molluscs from one of the lakes initially sampled, suggesting that these viruses are ubiquitous in freshwater environments. This study has attempted to generate baseline data of CRESS DNA viruses in certain environments using NGS-informed approaches. This data was used to try and establish whether viral distribution in different samples types can potentially be explained by the food web interactions between different samples types. Although the analysis did not show any significant relationships between sample type interactions and viral distribution a few common associations between Odonata larvae and benthic sediment were evident. This was expected as the larvae live within the sediment so it could be assumed that they potentially have similar CRESS DNA viral distribution. Although the distribution of viruses varied across sample types, molluscs proved the best sampling tool for isolating largest numbers of CRESS DNA viruses in an ecosystem with extensive diversity. Overall, this research demonstrates the applications of NGS for investigating the diversity of CRESS DNA viruses. It demonstrates that some sample types such as Odonata in terrestrial systems and molluscs in aquatic environments, can be used as effective sampling tool to determine the diversity of CRESS DNA viruses in different environments as well as detecting previously isolated viruses. The CRESS DNA viruses isolated in this body of work provides baseline data that can potentially be used in future research to investigate hosts of these viruses and their interactions with hosts and potential flow in their environments

Taxonomic research priorities for the conservation of the South African flora

Taxonomic revisions, monographs and floras are the most important, and often the only source of data for assessing the extinction risk of plants, with recent revisions contributing to more accurate assessments. The recently completed Red List of South African plants involved an overview of the taxonomic literature pertaining to the South African flora, providing an opportunity to identify critical gaps in taxonomic coverage. In this study we identified taxonomic research priorities for effective conservation of South African plants. Priorities were identified at genus level, according to time since last revision, level of endemism, collecting effort, proportion of taxa included in revisions, and specimen identification confidence. Although the results indicate that 62% of the flora has been recently revised, revisionary taxonomic output has declined drastically, particularly in the past 10 years. This decline is a result of a decrease in revisionary productivity per taxonomist and not a result of a decline in the number of working taxonomists. The family Aizoaceae is the top priority for taxonomic research with 55% of taxa in need of revision, followed by Hyacinthaceae with 34% of taxa not yet revised. Ericaceae, Euphorbiaceae, Rutaceae, Malvaceae, Asteraceae and Acanthaceae are also priorities with over 30% of taxa last revised before 1970. We recommend the reinstatement of the Flora of Southern Africa project in an online format in order to centralise South Africa’s existing taxonomic information and reinvigorate revisionary taxonomic study. This project will allow South Africa to fulfil its commitments to the Convention on Biodiversity by achieving Target 1 of the Global Strategy for Plant Conservation

Vegetation Map of South Africa, Lesotho and Swaziland 2009 and 2012: A description of changes from 2006

Background: The variety of applications in which the Vegetation Map of South Africa, Lesotho and Swaziland (VEGMAP) is used requires the map to be continually updated and refined to reflect the latest available information. The VEGMAP has been updated twice, in 2009 and 2012, since its first release in 2006. Objectives: The first objective is to report on the motivations for changes in the 2009 and 2012 versions. The second objective is to describe new vegetation types and subtypes included in these versions. Method: Changes to the VEGMAP are implemented after a peer-review process that is managed by the National Vegetation Map Committee. Accepted changes are then incorporated into the VEGMAP using GIS software. Results: Seventy-one of the 449 vegetation types were affected by updates. Changes included the addition of new vegetation types and subtypes, modifications to the boundaries of types present in the 2006 VEGMAP and changes to the names of vegetation types. Conclusion: The updates have affected a small portion of the map but have reflected a progressive refinement in quality. Regions that are still mapped at a coarse scale, especially those earmarked for land-use development, should be prioritised for improved map accuracy and classification through a more proactive approach towards vegetation mapping, using guidelines that are under development

Environmental Detection and Potential Transmission of Equine Herpesviruses

Equine herpesviruses (EHV) are a major health concern for domestic and wild equids and represent one of the most economically important disease agents of horses. Most known EHVs are transmitted directly between individuals as a result of direct exposure to exudates and aerosols. However, accumulating evidence suggests that environmental transmission may play a role including air, water, and fomites. Here, we reviewed studies on environmental stability and transmission of EHVs, which may influence viral dynamics and the use of environmental samples for monitoring EHV shedding.publishe

Evolutionary history of ssDNA bacilladnaviruses features horizontal acquisition of the capsid gene from ssRNA nodaviruses

International audienceBacilladnaviruses have single-stranded (ss) DNA genomes and infect diatoms, a major group of unicellular algae widespread in aquatic habitats. Despite their ecological importance, the provenance and relationships of bacilladnaviruses to other eukaryotic viruses remain unclear. Accordingly, they are currently classified into the 'floating' genus Bacilladnavirus. Here we present three new bacilladnavirus genomes recovered from a mollusc Amphibola crenata and benthic sediments from the Avon-Heathcote estuary in New Zealand. Our analysis shows that the rolling-circle replication-initiation proteins of bacilladnaviruses display unique conserved motifs and in phylogenetic trees form a monophyletic clade separated from other groups of ssDNA viruses. Unexpectedly, distant homology detection combined with structural modeling indicates that bacilladnavirus capsid proteins are homologous to those of ssRNA viruses from the Nodaviridae family. Considering the sequence diversity within the expanding Bacilladnavirus genus, we argue that classification of these viruses has to be revised and the current genus upgraded to the family level