Taxonomy and ecology of Botryosphaeria species and their anamorphs from Venezuela

Abstract

The fungal genus Botryosphaeria including its anamorphs has a cosmopolitan distribution and occurs on a wide range of monocotyledonous, dicotyledonous and gymnospermous hosts, including woody twigs and branches, herbaceous leaves, stems of grasses, and even lichen thalli. These fungi give rise to a wide variety of symptoms such as shoot blights, stem cankers, fruit rots, die-back and gummosis. They are also known as saprophytes on dead or dying stems, branches or leaves of plants. In Venezuela, the following species have been reported: Lasiodiplodia theobromae (the anamorph of Botryosphaeria rhodina), Diplodia pinea. D. mutila and Dothiorella spp. However, their characterization has been based only on morphological descriptions. The most common and well characterized species. L. theobromae, is associated with pines and other hosts. The focus of the studies presented in this thesis as to survey Eucalyptus and Acacia plantations in Venezuela for the presence and influence of Botryosphaeria spp., and to characterize these fungi using morphological characteristics and DNA sequence data. I also evaluated the pathogenicity and population biology of Botryosphaeria species present in the plantations. It was anticipated that the various studies would characterize a relatively large number of Botryosphaeria species, potentially recognize new species and provide some perspective of their relative importance to the Eucalyptus growing industry. The thesis has been divided into chapters that reflect discrete units prepared for future publication. The first chapter presents a review of the relevant literature. Two chapters treat the taxonomy of the Botryosphaeria spp. collected in this study. An additional two chapters consider the population biology of the most commonly encountered species and the remaining chapter deals with the pathogenicity of these fungi to Eucalyptus. In the literature review I treat the taxonomy of Botyosphaeria, which has been the subject of much uncertainty for many years. I also briefly review recent findings related to their molecular characterization. A focus is also placed on the pathogenicity of Botyosphaeria species on various host plants and their known importance in Venezuela. The focus concerns mainly Eucalyptus spp. Surveys that formed part of this study logically gave rise to a large collection of Botryosphaeria isolates. An important component of this thesis was to characterize these fungi. This was done based on morphological characteristics and also comparisons or DNA sequence data for various gene regions known to be informative for these fungi. Lasiodiplodia theobromae (anamorph of B.rhodina), B. ribis and B. parva were studied from a population biology perspective. These fungi were chosen for study because they were the most common species encountered on Eucalyptus in Venezuela. To study populations, I made use of simple sequence repeat markers (SS R). Three populations of each of L. theobromae (Venezuela, Mexico and South Africa) and B.ribis-B. parva complex (Venezuela, Colombia and Hawaii) were analysed and the data considered in terms of population differentiation, gene now, mode of reproduction, gene and genotype diversity. In the final chapter of this thesis, I evaluated the pathogenicity or the seven Botryosphaeria species identified from Venezuela. These seven species were thus inoculated on Eucalyptus urophylla x E. grandis hybrid clones. A second series of inoculations was then carried out with the two most pathogenic fungi to determine the relative tolerance or the most important clones to infection. This thesis was conducted over a period of four years. Work was undertaken both in Venezuela and South Africa and this necessitated long periods of time away from my home University and family. Surveys were conducted in many parts of Venezuela to collect the Botryospaeria spp. of interest for latter study in South Africa. The research chapters have been completed systematically and they evolved over time. Each represents a discrete unit implying that there is some overlap, at least in the references, between them. I would like to believe that the thesis will provide a firm foundation for further studies of Botryospaeria spp. and prove valuable to the small but important Eucalyptus growing industry in Venezuela.