Showing papers by "Michael J. Wingfield published in 2012"

Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi

Abstract: Six DNA regions were evaluated as potential DNA barcodes for Fungi, the second largest kingdom of eukaryotic life, by a multinational, multilaboratory consortium. The region of the mitochondrial cytochrome c oxidase subunit 1 used as the animal barcode was excluded as a potential marker, because it is difficult to amplify in fungi, often includes large introns, and can be insufficiently variable. Three subunits from the nuclear ribosomal RNA cistron were compared together with regions of three representative protein-coding genes (largest subunit of RNA polymerase II, second largest subunit of RNA polymerase II, and minichromosome maintenance protein). Although the protein-coding gene regions often had a higher percent of correct identification compared with ribosomal markers, low PCR amplification and sequencing success eliminated them as candidates for a universal fungal barcode. Among the regions of the ribosomal cistron, the internal transcribed spacer (ITS) region has the highest probability of successful identification for the broadest range of fungi, with the most clearly defined barcode gap between inter- and intraspecific variation. The nuclear ribosomal large subunit, a popular phylogenetic marker in certain groups, had superior species resolution in some taxonomic groups, such as the early diverging lineages and the ascomycete yeasts, but was otherwise slightly inferior to the ITS. The nuclear ribosomal small subunit has poor species-level resolution in fungi. ITS will be formally proposed for adoption as the primary fungal barcode marker to the Consortium for the Barcode of Life, with the possibility that supplementary barcodes may be developed for particular narrowly circumscribed taxonomic groups.

One fungus, one name promotes progressive plant pathology.

Abstract: The robust and reliable identification of fungi underpins virtually every element of plant pathology, from disease diagnosis to studies of biology, management/control, quarantine and, even more recently, comparative genomics. Most plant diseases are caused by fungi, typically pleomorphic organisms, for which the taxonomy and, in particular, a dual nomenclature system have frustrated and confused practitioners of plant pathology. The emergence of DNA sequencing has revealed cryptic taxa and revolutionized our understanding of relationships in the fungi. The impacts on plant pathology at every level are already immense and will continue to grow rapidly as new DNA sequencing technologies continue to emerge. DNA sequence comparisons, used to resolve a dual nomenclature problem for the first time only 19 years ago, have made it possible to approach a natural classification for the fungi and to abandon the confusing dual nomenclature system. The journey to a one fungus, one name taxonomic reality has been long and arduous, but its time has come. This will inevitably have a positive impact on plant pathology, plant pathologists and future students of this hugely important discipline on which the world depends for food security and plant health in general. This contemporary review highlights the problems of a dual nomenclature, especially its impact on plant pathogenic fungi, and charts the road to a one fungus, one name system that is rapidly drawing near.

Retracing the routes of introduction of invasive species: the case of the Sirex noctilio woodwasp

Abstract: Understanding the evolutionary histories of invasive species is critical to adopt appropriate management strategies, but this process can be exceedingly complex to unravel. As illustrated in thisstudy ofthe worldwide invasion of thewoodwaspSirex noctilio, population genetic analyses using coalescent-based scenario testing together with Bayesian clustering and historical records provide opportunities to address this problem. The pest spread from its native Eurasian range to the Southern Hemisphere in the 1900s and recently to Northern America, where it poses economic and potentially ecological threats to planted and native Pinus spp. To investigate the origins and pathways of invasion, samples from five continents were analysed using microsatellite and sequence data. The results of clustering analysis and scenario testing suggest that the invasion history is much more complex than previously believed, with most of the populations being admixtures resulting from independent introductions from Europe and subsequent spread among the invaded areas. Clustering analyses revealed two major source gene pools, one of which the scenario testing suggests is an as yet unsampled source. Results also shed light on the microevolutionary processes occurring during introductions, and showed that only few specimens gave rise to some of the populations. Analyses of microsatellites using clustering and scenario testing considered against historical data drastically altered our understanding of the invasion history of S. noctilio and will have important implications for the strategies employed to fight its spread. This study illustrates the value of combining clustering and ABC methods in a comprehensive framework to dissect the complex patterns of spread of global invaders.

Fungal Planet description sheets: 128-153

Abstract: Novel species of microfungi described in the present study include the following from Australia: Catenulostroma corymbiae from Corymbia, Devriesia stirlingiae from Stirlingia, Penidiella carpentariae from Carpentaria, Phaeococcomyces eucalypti from Eucalyptus, Phialophora livistonae from Livistona, Phyllosticta aristolochiicola from Aristolochia, Clitopilus austroprunulus on sclerophyll forest litter of Eucalyptus regnans and Toxicocladosporium posoqueriae from Posoqueria. Several species are also described from South Africa, namely: Ceramothyrium podocarpi from Podocarpus, Cercospora chrysanthemoides from Chrysanthemoides, Devriesia shakazului from Aloe, Penidiella drakensbergensis from Protea, Strelitziana cliviae from Clivia and Zasmidium syzygii from Syzygium. Other species include Bipolaris microstegii from Microstegium and Synchaetomella acerina from Acer (USA), Brunneiapiospora austropalmicola from Rhopalostylis (New Zealand), Calonectria pentaseptata from Eucalyptus and Macadamia (Vietnam), Ceramothyrium melastoma from Melastoma (Indonesia), Collembolispora aristata from stream foam (Czech Republic), Devriesia imbrexigena from glazed decorative tiles (Portugal), Microcyclospora rhoicola from Rhus (Canada), Seiridium phylicae from Phylica (Tristan de Cunha, Inaccessible Island), Passalora lobeliaefistulosis from Lobelia (Brazil) and Zymoseptoria verkleyi from Poa (The Netherlands). Valsalnicola represents a new ascomycete genus from Alnus (Austria) and Parapenidiella a new hyphomycete genus from Eucalyptus (Australia). Morphological and culture characteristics along with ITS DNA barcodes are also provided.

The sirex woodwasp and its fungal symbiont : research and management of a worldwide invasive pest

Abstract: Preface Dedication 1.0 Sirex systematics problems and solutions Henri Goulet 1.1 Introduction 1.2 Important issues for solving identification problems 1.2.1 Ovipositor morphology 1.2.2 Mitochondrial DNA 1.2.3 Collections 1.3 Diversity and biogeographic coverage 1.4 Morphological characters used to recognize species 1.4.1 Ovipositor characters 1.4.2 Other characters 2.0 Life history and biology of Sirex noctilio Kathleen Ryan and Brett P Hurley 2.1 Introduction 2.2 Insect-fungus complex 2.3 The life history of Sirex noctilio 2.4 Host selection and susceptibility 2.5 Host Records 2.6 Factors influencing population dynamics 2.6.1 Potential fecundity 2.6.2 Reproductive potential 2.6.3 Adult size 2.6.4 Sex-ratio 2.6.5 Natural enemies 2.7 Conclusions 3.0 Susceptibility and response of pines to Sirex noctilio John M Bordeaux and Jeffrey FD Dean 3.1 Introduction 3.2 Factors contributing to tree susceptibility 3.3 Tree responses to Sirex noctilio mucus 3.4 Tree responses to Amylostereum areolatum 3.5 General responses to Sirex attack 3.6 Reducing tree susceptibility 4.0 The population dynamics of Sirex noctilio F.: the influence of diapause, spatial aggregation and flight potential on woodwasp outbreaks and spread Juan C Corley and Jose M Villacide 4.1 Introduction 4.2 Pine plantations and Sirex noctilio in Patagonia 4.3 Life cycle and adult emergence patterns 4.4 Spatial dynamics of woodwasp populations at a stand scale 4.5 Flight potential of Sirex noctilio males and females 4.6 Conclusions 5.0 The woodwasp Sirex noctilio and its associated fungus Amylostereum areolatum in Europe Beat Wermelinger and Iben M Thomsen 5.1 Introduction 5.2 Biology of Sirex noctilio 5.3 Natural enemies 5.4 Ecology of the Amylostereum fungal symbionts 5.5 Relationships between S. noctilio attack and host quality 5.6 Economic significance of S. noctilio in Europe 6.0 The genetics of Amylostereum species associated with Siricids Magriet van der Nest, Rimvys Vasaitis, Jan Stenlid, Brenda D Wingfield, Michael J Wingfield and Bernard Slippers 6.1 Introduction 6.2 Systematics and identification 6.3 Symbiotic specificity 6.4 Mode of reproduction 6.5 Vegetative incompatibility and population diversity studies 6.6 Conclusions and future prospects 7.0 Siricid woodwasps and their fungal symbionts in Asia, specifically those occurring in Japan Masanobu Tabata, Hiroaki Miyata and Kaoru Maeto 7.1 Introduction 7.2 Symbiosis between woodwasps and fungi 7.3 Molecular phylogeny of fungal symbionts 7.4 Life-cycle and host interaction of woodwasps 8.0 Parasitoids in the management of Sirex: Looking back and looking ahead

Biological control of forest plantation pests in an interconnected world requires greater international focus

Abstract: The worldwide homogenization of genetic resources used in plantation forestry (primarily Pinus, Eucalypus, Populus and Acacia spp.) together with accelerating rates of human-aided dispersal of exotic pests, is resulting in plantation pests becoming broadly distributed extremely quickly, sometimes reaching a global distribution within a decade. This unprecedented rate of establishment and spread means that the risk associated with new and emerging pests is shared globally. Biological control represents a major component of the strategy to mitigate such risk, but the current efforts and scope for developing such controls are woefully inadequate for dealing with the increasing rates of pest spread. Given the global nature of the problem, biological control would benefit enormously from an international, collaborative focus. Though inherent difficulties and potential pitfalls exist, opportunities for cost-sharing, growth and maintenance of resources and capacity, and more comprehensive research programmes are...

Phylogeny and taxonomy of species in the Grosmannia serpens complex.

Abstract: Grosmannia serpens was first described from pine in Italy in 1936 and it has been recorded subsequently from many countries in both the northern and southern hemispheres. The fungus is vectored primarily by root-infesting bark beetles and has been reported to contribute to pine-root diseases in Italy and South Africa. The objective of this study was to consider the identity of a global collection of isolates not previously available and using DNA sequence-based comparisons not previously applied to most of these isolates. Phylogenetic analyses of the ITS2-LSU, actin, beta-tubulin, calmodulin and translation elongation factor-1 alpha sequences revealed that these morphologically similar isolates represent a complex of five cryptic species. Grosmannia serpens sensu stricto thus is redefined and comprises only isolates from Italy including the ex-type isolate. The ex-type isolate of Verticicladiella alacris was shown to be distinct from G. serpens, and a new holomorphic species, G. alacris, is described. The teleomorph state of G. alacris was obtained through mating studies in the laboratory, confirming that this species is heterothallic. Most of the available isolates, including those from South Africa, USA, France, Portugal and some from Spain, represent G. alacris. The remaining three taxa, known only in their anamorph states, are described as the new species Leptographium gibbsii for isolates from the UK, L. yamaokae for isolates from Japan and L. castellanum for isolates from Spain and the Dominican Republic.

Associations of Conifer-Infesting Bark Beetles and Fungi in Fennoscandia.

Abstract: Bark beetles (Coleoptera, Scolytinae) have a widespread association with fungi, especially with ophiostomatoid fungi (Ascomycota) that cause blue staining of wood, and in some cases, serious tree diseases. In Fennoscandia, most studies of these fungi have focused on economically important bark beetle species and this is likely to have led to a biased view of the fungal biodiversity in the region. Recently, the associations between fungi and bark beetles in Fennoscandia have been shown to be more diverse than previously thought. Furthermore, they form complex and dynamic associations that are only now beginning to emerge. This review examines the current knowledge of the rather poorly known interactions between bark beetles, fungi and their conifer host trees in Fennoscandia. The diversity of ophiostomatoid species is discussed and the possible factors that influence the assemblages of fungal associates are considered for all species that are known to occur in the region. For many ophiostomatoid species found in Fennoscandia, little or nothing is known regarding their pathogenicity, particularly if they were to be transferred to new environments. We, therefore, draw attention to the possible threats of timber trade and climate change-induced invasions of new habitats by bark beetles and the fungi that can be moved along with them.

Diversity in Eucalyptus susceptibility to the gall‐forming wasp Leptocybe invasa

Abstract: Extensive variation to damage by the invasive gall-forming wasp Leptocybe invasa Fisher & LaSalle (Hymenoptera: Eulophidae) is known to exist amongst Eucalyptus genotypes. 2 In the present study, 30 of the 50 tested genotypes were susceptible to gall formation and development of the wasp. Gall development on the petiole and leaves of plants was compared to calculate the percentage of infestation per plant and per genotype. 3 A positive correlation between galls on petioles and leaves indicated an absence of specificity at this level, and also that either leaves or petioles could be used to obtain an accurate estimate of the level of infestation. 4 Genotypes of E. nitens × E. grandis and E. grandis × E. camaldulensis were most susceptible, with a maximum damage index value for leaves and petioles of 0.52 and 0.39, respectively. Eucalyptus dunii, E. nitens, E. smithii, E. urophylla and E. saligna × E. urophylla showed little or no infestation. 5 The results obtained in the present study suggest that the selection and planting of resistant/less susceptible genotypes will be an important aid in managing damage from L. invasa invasion.

First fungal genome sequence from Africa: A preliminary analysis

Abstract: Some of the most significant breakthroughs in the biological sciences this century will emerge from the development of next generation sequencing technologies. The ease of availability of DNA sequence made possible through these new technologies has given researchers opportunities to study organisms in a manner that was not possible with Sanger sequencing. Scientists will, therefore, need to embrace genomics, as well as develop and nurture the human capacity to sequence genomes and utilise the ’tsunami‘ of data that emerge from genome sequencing. In response to these challenges, we sequenced the genome of Fusarium circinatum, a fungal pathogen of pine that causes pitch canker, a disease of great concern to the South African forestry industry. The sequencing work was conducted in South Africa, making F. circinatum the first eukaryotic organism for which the complete genome has been sequenced locally. Here we report on the process that was followed to sequence, assemble and perform a preliminary characterisation of the genome. Furthermore, details of the computer annotation and manual curation of this genome are presented. The F. circinatum genome was found to be nearly 44 million bases in size, which is similar to that of four other Fusarium genomes that have been sequenced elsewhere. The genome contains just over 15 000 open reading frames, which is less than that of the related species, Fusarium oxysporum, but more than that for Fusarium verticillioides. Amongst the various putative gene clusters identified in F. circinatum, those encoding the secondary metabolites fumosin and fusarin appeared to harbour evidence of gene translocation. It is anticipated that similar comparisons of other loci will provide insights into the genetic basis for pathogenicity of the pitch canker pathogen. Perhaps more importantly, this project has engaged a relatively large group of scientists including students in a significant genome project that is certain to provide a platform for growth in this important area of research in the future.

Grosmannia and Leptographium spp. associated with conifer-infesting bark beetles in Finland and Russia, including Leptographium taigense sp. nov.

Abstract: Species of Grosmannia with Leptographium anamorphs include important forest pathogens and agents of blue stain in timber. They are commonly found in association with forest pests, such as bark beetles. During a survey of ophiostomatoid fungi in eastern parts of Finland and neighboring Russia, species belonging to the genus Grosmannia were isolated from 12 different bark beetle species infesting Picea abies and Pinus sylvestris, the most economically important conifers in the region. Identification of these fungi was based on morphology, DNA sequence comparisons for three gene regions and phylogenetic analyses. A total of ten taxa were identified. These belonged to six different species complexes in Grosmannia. The phylogenetic analyses provided an opportunity to redefine the G. galeiformis-, L. procerum-, L. lundbergii-, G. piceiperda-, G. olivacea- and G. penicillata-complexes, and to consider the species emerging from the survey within the context of these complexes. The species included G. galeiformis, G. olivacea, L. chlamydatum, L. lundbergii, L. truncatum and a novel taxon, described here as L. taigense sp. nov. In addition, species closely related to G. cucullata, G. olivaceapini comb. nov., G. piceiperda and L. procerum were isolated but their identity could not be resolved. The overall results indicate that the diversity of Grosmannia species in the boreal forests remains poorly understood and that further studies are needed to clarify the status of several species or species complexes.

First fungal genome sequence from Africa : a preliminary analysis : research article

Abstract: Some of the most significant breakthroughs in the biological sciences this century will emerge from the development of next generation sequencing technologies. The ease of availability of DNA sequence made possible through these new technologies has given researchers opportunities to study organisms in a manner that was not possible with Sanger sequencing. Scientists will, therefore, need to embrace genomics, as well as develop and nurture the human capacity to sequence genomes and utilise the 'tsunami' of data that emerge from genome sequencing. In response to these challenges, we sequenced the genome of Fusarium circinatum, a fungal pathogen of pine that causes pitch canker, a disease of great concern to the South African forestry industry. The sequencing work was conducted in South Africa, making F. circinatum the first eukaryotic organism for which the complete genome has been sequenced locally. Here we report on the process that was followed to sequence, assemble and perform a preliminary characterisation of the genome. Furthermore, details of the computer annotation and manual curation of this genome are presented. The F. circinatum genome was found to be nearly 44 million bases in size, which is similar to that of four other Fusarium genomes that have been sequenced elsewhere. The genome contains just over 15 000 open reading frames, which is less than that of the related species, Fusarium oxysporum, but more than that for Fusarium verticillioides. Amongst the various putative gene clusters identified in F. circinatum, those encoding the secondary metabolites fumosin and fusarin appeared to harbour evidence of gene translocation. It is anticipated that similar comparisons of other loci will provide insights into the genetic basis for pathogenicity of the pitch canker pathogen. Perhaps more importantly, this project has engaged a relatively large group of scientists including students in a significant genome project that is certain to provide a platform for growth in this important area of research in the future.

Fusarium circinatum and pitch canker of Pinus in Colombia

Abstract: Pitch canker, caused by the ascomycete fungus Fusarium circinatum, infects a wide range of Pinus species. The pathogen has a global distribution and limits plantation productivity wherever susceptible Pinus species are commercially cultivated. During 2005–2007, symptoms typical of those associated with F. circinatum were observed in Colombia on nursery seedlings of P. maximinoi, P. tecunumanii and P. patula, as well as established P. patula and P. kesiya trees in plantations. Symptoms on seedlings included collar and root disease while shoot dieback and resinous stem cankers were found on trees in plantations. The aim of this study was to isolate and identify the causal agent of these symptoms and to evaluate the relative tolerance of various families of Pinus species commonly grown in Colombia. By making use of morphology and DNA-based methods, as well as pathogenicity tests on P. patula seedlings, it was possible to show that the symptoms observed in the nursery and field were caused by F. circinatum. Furthermore, the results of pathogenicity tests with two virulent isolates of the pathogen indicated that P. tecunumanii from low-elevation sources and P. maximinoi are significantly more tolerant to infection by F. circinatum than P. tecunumanii from high-elevation sources and P. patula. These results show that there is substantial opportunity to avoid losses due to infection by F. circinatum through deployment of resistant planting stock.

Ceratocystis eucalypticola sp. nov. from Eucalyptus in South Africa and comparison to global isolates from this tree

Abstract: Eucalyptus trees, mostly native to Australia, are widely planted in the tropics and Southern Hemisphere for the production of wood and pulp. Worldwide surveys of diseases on these trees have yielded a large collection of Ceratocystis isolates from dying trees or from wounds on their stems. The aim of this study was to characterise these isolates and to consider their relatedness to each other. Culture appearance, morphological features and a distinctive fruity odour in all cultures were typical of species in the Ceratocystis fimbriata sensu lato (s. lat.) complex. Phylogenetic analyses of sequences for the combined ITS, βt-1 and TEF1-α gene regions revealed a genetically diverse group of isolates residing in a single large clade, that were distinct from all other species in the C. fimbriata s. lat. complex. Based on morphology and phylogenetic inference, the Eucalyptus isolates are recognised as closely related. The South African isolates are described here as a new species, C. eucalypticola.

Ceratocystis species, including two new species associated with nitidulid beetles, on eucalypts in Australia

Abstract: The genus Ceratocystis includes important fungal pathogens of trees, including Eucalyptus spp Ironically, very little is known regarding the diversity or biology of Ceratocystis species on Eucalyptus species in Australia, where most of these trees are native The aim of this study was to survey for Ceratocystis spp, and their possible insect associates, on eucalypts in Australia and thus to establish a foundation of knowledge regarding these fungi on the continent Collections were made in three states of Australia from wounds on trees, as well as from nitidulid beetles associated with these wounds Ceratocystis spp were identified based on morphology and multigene sequence comparisons Of the 54 isolates obtained, two previously unknown species of Ceratocystis were found and these are described here as Ceratocystis corymbiicola sp nov and Ceratocystis tyalla sp nov Furthermore, the distribution of Ceratocystis pirilliformis is expanded to include Eucalyptus spp in Tasmania

Cryptic species, native populations and biological invasions by a eucalypt forest pathogen

Abstract: Human-associated introduction of pathogens and consequent invasions is very evident in areas where no related organisms existed before. In areas where related but distinct populations or closely related cryptic species already exist, the invasion process is much harder to unravel. In this study, the population structure of the Eucalyptus leaf pathogen Teratosphaeria nubilosa was studied within its native range in Australia, including both commercial plantations and native forests. A collection of 521 isolates from across its distribution was characterized using eight microsatellite loci, resulting in 112 multilocus haplotypes (MLHs). Multivariate and Bayesian analyses of the population conducted in structure revealed three genetically isolated groups (A, B and C), with no evidence for recombination or hybridization among groups, even when they co-occur in the same plantation. DNA sequence data of the ITS (n = 32), β-tubulin (n = 32) and 27 anonymous loci (n = 16) were consistent with microsatellite data in suggesting that T. nubilosa should be considered as a species complex. Patterns of genetic diversity provided evidence of biological invasions by the pathogen within Australia in the states of Western Australia and New South Wales and helped unravel the pattern of invasion beyond Australia into New Zealand, Brazil and Uruguay. No significant genetic differences in pathogen populations collected in native forests and commercial plantations were observed. This emphasizes the importance of sanitation in the acquisition of nursery stock for the establishment of commercial plantations.

Factors Affecting the Efficacy of Deladenus siricidicola in Biological Control Systems

Abstract: The nematode, Deladenus (=Beddingia) siricidicola, represents the cornerstone of Sirex noctilio biological control programs across the Southern Hemisphere. There is, however, significant variation in its efficacy in different regions. In this review, we consider emerging issues related to the biology and handling of the nematode that might influence its efficacy in biological control systems. Most practical aspects concerning the handling of D. siricidicola have been streamlined over the past half-century and these appear to be very efficient. However, large gaps remain in our knowledge about some key aspects of the biology of D. siricidicola. For example, very little is known regarding the evolution of virulence in the nematode populations, and the consequent evolution of resistance in S. noctilio populations. Furthermore, the levels of diversity in D. siricidicola and its ability to adapt to fungal, wasp and environmental variation are poorly understood. In this regard, new collections and storage of native populations of the Deladenus spp. are critical for the future research and management of this key biological control agent of S. noctilio.

Selection of Pinus spp. in South Africa for tolerance to infection by the pitch canker fungus

Abstract: The increasing threats from pests and diseases demand that the South African forest industry explores options to deploy alternative pine species in plantation development. This is especially true for species, such as Pinus patula Schiede and Deppe ex Schltdl. and Cham., which are highly susceptible to the pitch canker fungus Fusarium circinatum. Losses due to F. circinatum have been confined mostly to nurseries and at field establishment resulting in a significant cost to the industry. Although, the fungus has not as yet resulted in stem and branch infections on established P. patula in South Africa, it has caused pitch canker on other, more susceptible species such as P. radiata D. Don., and P. greggii Engelm. ex Parl. As alternatives to P. patula, on the warmer and cooler sites in South Africa, families of P. elliottii Engelm var. elliottii, P. tecunumanii (Schw.) Eguiluz and Perry, P. maximinoi H. E. Moore and P. pseudostrobus Lindl. were screened for tolerance to infection by F. circinatum in greenhouse studies. Seedlings were wounded and inoculated with spores of F. circinatum. Lesion development following inoculation was used to differentiate the levels of tolerance between families. The results showed that P. maximinoi, P. pseudostrobus, and the low elevation variety of P. tecunumanii are highly tolerant to infection with very little family variation. The narrow sense heritability estimates for these species were less than 0.06. In contrast, P. elliottii showed good tolerance with some family variation and a heritability of 0.22, while the high elevation source of P. tecunumanii showed a high degree of family variation and a heritability of 0.59. These results provide the industry with valuable information on pine species tolerant to F. circinatum that could be used as alternatives to P. patula in South Africa.

DNA extraction techniques for DNA barcoding of minute gall-inhabiting wasps.

Abstract: DNA extraction from minute hymenopterans and their larvae is difficult and challenging because of their small size indicating a low amount of starting material. Hence, 11 DNA extraction methods were compared to determine their efficacy in isolating DNA. Success of each method was scored on a 2% agarose gel after PCR of the cox 1 mitochondrial locus. A silica-membrane-based approach was the most successful, followed by a method using a combination of incubation buffers and a method using magnetic beads. The method using buffers was the most cost- and time effective. Using this method, larvae from Eucalyptus seed capsule galls could be assigned a role (parasitoid, gall former or inquiline) in the gall-inhabiting complex.

The Control of the Sirex Woodwasp in Diverse Environments: The South African Experience

Abstract: The woodwasp Sirex noctilio was detected in South Africa in 1994, in Cape Town. Within the following 15 years, S. noctilio had spread across four of the five main forestry regions. Losses incurred from S. noctilio damage increased dramatically as it moved into the Eastern Cape and KwaZulu-Natal provinces, where densely stocked pulpwood stands provided ideal conditions for the population of the wasp to increase. Control strategies developed in Australasia were adopted, including the introduction of the parasitic nematode Deladenus (=Beddingia) siricidicola and the parasitic wasp Ibalia leucospoides. Adaptations were required for the techniques used to release D. siricidicola due to the poor success obtained in the summer rainfall areas. These adaptations increased the effectiveness of the inoculations and subsequent natural spread, although overall inoculation success remained poor. Ibalia leucospoides was successfully released in the Western Cape in 1998, but only recently (since 2006) have there been efforts to release this parasitoid in other infested areas. Monitoring has been crucial for the early detection of infestations and release of biological control agents. Plantation management strategies to combat S. noctilio have been used to some extent, but are restricted by economic considerations. Although infestations of S. noctilio have recently decreased, the movement of S. noctilio into the largest pine-growing province and the current inability to quickly establish biological control due to poor inoculation success emphasizes the fact that S. noctilio continues to pose a serious threat to the pine industry in South Africa.

The potential for monitoring and control of insect pests in Southern Hemisphere forestry plantations using semiochemicals

Abstract: Southern Hemisphere plantation forestry has grown substantially over the past few decades and will play an increasing role in fibre production and carbon sequestration in future. The sustainability of these plantations is, however, increasingly under pressure from introduced pests. This pressure requires an urgent and matching increase in the speed and efficiency at which tools are developed to monitor and control these pests. To consider the potential role of semiochemicals to address the need for more efficient pest control in Southern Hemisphere plantations, particularly by drawing from research in other parts of the world. Semiochemical research in forestry has grown exponentially over the last 40 years but has been almost exclusively focussed on Northern Hemisphere forests. In these forests, semiochemicals have played an important role to enhance the efficiency of integrated pest management programmes. An analysis of semiochemical research from 1970 to 2010 showed a rapid increase over time. It also indicated that pheromones have been the most extensively studied type of semiochemical in forestry, contributing to 92% of the semiochemical literature over this period, compared with research on plant kairomones. This research has led to numerous applications in detection of new invasions, monitoring population levels and spread, in addition to controlling pests by mass trapping or disrupting of aggregation and mating signals. The value of semiochemicals as an environmentally benign and efficient approach to managing forest plantation pests in the Southern Hemisphere seems obvious. There is, however, a lack of research capacity and focus to optimally capture this opportunity. Given the pressure from increasing numbers of pests and reduced opportunities to use pesticides, there is some urgency to develop semiochemical research capacity.

A diverse assemblage of Ophiostoma species, including two new taxa on eucalypt trees in South Africa

Abstract: Fungi in the Ophiostomatales include important pathogens of trees as well as agents of wood stain, reducing the economic value of timber. They rely on insects, such as bark beetles, for dispersal and are commonly associated with wounds on trees. Although Ophiostoma spp. have been reported from eucalypt wood chips in South Africa, very little is known about the diversity of the Ophiostomatales, or their insect associates, on plantation-grown Eucalyptus spp. The aim of this study was to consider the diversity and distribution of the Ophiostomatales infecting fresh wounds on Eucalyptus trees in the country. Additionally, knowledge regarding their association with nitidulid beetles, which have previously been shown to carry Ophiostoma spp., was sought. Surveys were conducted in five provinces where Eucalyptus spp. are commonly grown, and the fungi collected were identified using morphological comparisons and multigene sequence phylogenies. Of the 139 isolates collected, five Ophiostoma spp. were identified including O. quercus, O. tsotsi and O. tasmaniense. These were from cut stumps as well as from the nitidulid beetles Brachypeplus depressus and Carpophilus spp. In addition, two new taxa in the O. stenoceras–Sporothrix schenckii complex were identified from Eucalyptus trees infested by Phoracantha semipunctata. The two new taxa are described as O. candidum sp. nov., and O. fumeum sp. nov., respectively. The results of this study clearly show that the diversity and ecology of Ophiostoma spp. on Eucalyptus trees in South Africa is poorly understood and that further studies are required to determine the possible economic relevance of these fungi.

Future outlook for Pinus patula in South Africa in the presence of the pitch canker fungus (Fusarium circinatum)

Abstract: Approximately 50% of the area planted to softwood trees in South Africa has been established with Pinus patula, making it the most important pine species in the country. More effort has gone into developing this species for improved growth, tree form and wood properties than with any other species. This substantial investment has been threatened in the last 10 years by the pitch canker fungus, Fusarium circinatum. The fungus infects and contaminates nursery plants and, once transferred to the field, causes severe mortality of young trees in the first year after establishment. Although nurserymen have some control of the disease, it is recognised that the best long-term solution to mitigate damage because of F. circinatum infection is to identify tolerant species, clones and hybrids for deployment in plantations in the future. Research has shown that alternative species such as P. tecunumanii, P. maximinoi and P. elliottii are suitable for warm sites. Pine hybrids, particularly between P. patula and the hi...

High levels of genetic diversity and cryptic recombination is widespread in introduced Diplodia pinea populations

Abstract: Introduced populations of organisms typically have reduced diversity compared to those that are native. It is, therefore, unusual that introduced populations of the fungal tree pathogen Diplodia pinea have been shown to have high levels of genetic diversity, even surpassing diversity in some native regions. This is thought to be due to multiple introductions over time or the existence of a cryptic and yet undiscovered sexual cycle. In this study, we consider whether populations of D. pinea in Southern Hemisphere countries have similar patterns of diversity, share some level of genetic identity and how they might be influenced by sexual recombination. A total of 173 isolates from Argentina, Australia, Ethiopia and South Africa were characterized using 12 microsatellite markers. The results show that all these populations have high gene and genotype diversities, with the Australian population having the lowest diversity. Very few private alleles were found, suggesting that isolates from different countries might share a source of introduction. However, based on allele distribution and frequency, each of the populations appeared to be evolving independently. The results showed that in all but the Australian population, alleles are randomly associated, suggesting that widespread sexual recombination has influenced population structure.