Phytophthora nicotianae on tobacco and its control in South Africa

Abstract

As the causative agent of black shank, Phytophthora nicotianae is a serious threat to tobacco cultivation in South Africa. Research presented in this dissertation describes pathogenicity studies and control measures for P. nicotianae on tobacco. Special attention is given to the population structure of P. nicotianae in South Africa. The implications of these genetic studies in breeding and selection programs against P. nicotianae were also evaluated. The first chapter of this dissertation represents a literature review on black shank and available control measures for P. nicotianae on tobacco. The mechanisms of pathogenicity and the life cycle of P. nicotianae are also treated in detail. Special reference is made to the maintenance of genetic diversity in Phytophthora species and particularly P. nicotianae. This literature review also highlights the fact that very few studies have been conducted to determine the genetic structure of P. nicotianae populations. The success of South African breeding programs for tobacco cultivars with P. nicotianae resistance is to some degree dependent on the selection of isolates with high levels of aggressiveness. The research presented in chapter two provides information on cultivar resistance and selection of P. nicotianae isolates for future breeding programs. Significant differences in levels of aggressiveness were found between P. nicotianae isolates. Furthermore, race 0 and 1 of P. nicotianae occurred in most of the tobacco growing regions in South Africa. Selected Race 0 and 1 isolates were thus used to evaluate black shank resistance of 11 commercially planted tobacco cultivars. Commercially planted cultivars differed significantly in their resistance to race 0 and 1. Cultivars LK33/60 and OD1 were highly resistant to race 0 but susceptible to race 1 while cultivars Vuma/3/46 and LK3/46 were highly resistant to both race 0 and 1. Chapter three reports on the use of metalaxyl treatments combined with resistance in tobacco cultivars for control of P. nicotianae. One hundred and thirty two isolates of P. nicotianae were screened for sensitivity to metalaxyl. P. nicotianae isolates from most tobacco farms were metalaxyl sensitive. The results further indicated that the use of metalaxyl in combination with moderately resistant cultivars effectively reduced black shank in the field. The outcome of this study provided useful information for the implementation of an economically viable combination of disease resistance and metalaxyl as the basis for a P. nicotianae management program in South Africa. Chapter four of this dissertation deals with the development of a rapid seedling-' based screening technique to assay tobacco for resistance to P. nicotianae. This technique was validated by comparing it to a stem inoculation technique commonly used on adult plants. A strong positive correlation was found between results of the seedling assay and adult plant trials for all isolates and cultivars tested. P. nicotianae isolates could also be characterized as race 0 or I using both stem inoculation and the rapid seedling assay. The ability to screen large numbers of tobacco plants rapidly at the seedling stage allows for the testing of large germplasm resources in a systematic manner and under standard conditions. This may help in the timely development and release of more black shank resistant cultivars. In chapter five, a population study on P. nicotianae in South Africa is presented. One hundred and five P. nicotianae isolates were collected from the Northern Highveld and Lowveld regions, as well as from both citrus and tobacco hosts in South Africa. Levels of phenotypic diversity were determined in populations of P. nicotianae using RAPD markers. Among the 105 P. nicotianae isolates analysed 79 different RAPD phenotypes were found, where 35 of the isolates were found to be clonal. The high number of RAPD phenotypes (79) in relation to the sample size (105), the presence of both the Al and A2 mating type and high levels of phenotypic diversity in the P. nicotianae population indicate a sexually outcrossing P. nicotianae population in South Africa. This sexual outcrossing may mean that P. nicotianae is likely to remain a constant threat to tobacco and citrus cultivation, since new genotypes with the potential to overcome resistance genes in commercial cultivars are likely to emerge. All chapters of this dissertation deal with some aspects of black shank control and breeding for resistance to P. nicotianae. This dissertation provides new knowledge on variation in levels of aggressiveness, race distribution and the development of metalaxyl resistance in the South African P. nicotianae populations. This also represents the first study on the genetic diversity of P. nicotianae populations in South Africa. The results presented here not only show the possible occurrence of sexual reproduction, but also indicate the presence of clones and discreet phenotypic groups of P. nicotianae. This information will be applied in future tobacco breeding programs to select breeding lines with resistance against a number of specific P. nicotianae races and phenotypic groups.