Student Number : 0413249D - MSc research report - Faculty of Science / Cassava is one of the most important staple crops in the world and is consumed by over 700 million people around the globe and is a profitable product commercially due to the high starch content of its tubers. One of the future aims is to produce cassava that is high yielding, resistant to cassava mosaic geminiviruses (CMGs) and high in starch content. To be able to achieve commercially attractive cassava varieties, research need to be carried out to investigate the virus resistance status of different cassava cultivars, which can later be used in the future breeding programme. In South Africa, cassava is used for commercial starch manufacturing purposes, as a cash crop and a food source by small-scale farmers. Cassava Mosaic Disease (CMD) is having a negative impact on yield of the crop globally and therefore dropping profitability of cassava on a commercial scale. The aims of this research were to propagate thirteen cassava cultivars and then to test them for virus susceptibility or resistance. Eleven cassava cultivars received from the International Institute of Tropical Agriculture (IITA) were tested for resistance or susceptibility against South African cassava mosaic virus (SACMV). Two local, commercial cultivars T200 and T400, were tested for East African cassava mosaic virus (EACMV) and African cassava mosaic virus (ACMV) resistance. Cassava cultivars were successfully propagated in vitro and thereafter transferred into soil and acclimatized to adapt to environmental conditions. When the plantlets were three weeks old, the plantlets were infected with cassava mosaic viruses. Plants were infected with SACMV via Agrobacterium-mediated transfer and infectious EACMV and ACMV monomers were used to biolistically bombard the plantlets. Resistance/susceptibility results of seven of the thirteen cultivars were obtained with SACMV, these cultivars being T200 (susceptible), T400 (susceptible), TME3 (highly resistant), I30572 (susceptible), I420251 (highly susceptible), I60506 (susceptible) and TMS60444 (susceptible). Due to destruction by fungal gnats eating the roots of the plants, acclimatization of the remaining six cultivars was not possible. Also, due to the nature of the biolistic equipment, infection of the cultivars with EACMV and ACMV was not achieved as the plantlets were not robust enough to survive the pressure.

Cassava (Manihot esculenta Crantz) is a basic staple food crop in Tanzania. Cassava mosaic disease (CMD) caused by cassava mosaic geminiviruses (CMGs) constitutes a major limiting factor to cassava production in the country. This study was undertaken to characterize the CMGs occurring in Tanzania using molecular techniques and to map their geographical distribution to generate information on which the formulation of control measures can be based. Using Polymerase Chain Reaction (PCR) and Restriction Fragment Length Polymorphism (RFLP) for analysis of CMGs DNA-A genomes, different CMGs were found to be associated with CMD. Higher molecular diversity was observed among East African cassava mosaic viruses (EACMVs) than African cassava mosaic viruses (ACMVs), which was confirmed later by complete nucleotide sequence analysis. In addition to EACMV and ACMV isolates, two isolates of EACMV Cameroon virus (EACMCV) were found in Tanzania. These were confirmed to be strains of EACMCV Cameroon, originally described in Cameroon, West Africa and here named EACMCV- [TZ1] and EACMCV-[TZ7]. They had high (92%) overall DNA-A nucleotide sequence identity and EACMCV-[TZ1] was widespread in the southern part of the country. A subgenomic DNA form of CMG that appeared to be truncated was identified in a CMD-infected cassava plant. It was confirmed upon sequence analysis to be a defect of EACMV DNA-A and had a capacity of attenuating symptoms when coinoculated with wild-type EACMV. In addition, this study revealed for the first time the presence of two novel non-geminivirus single-stranded DNA (ssDNA) sub-genomic molecules associated with CMG infection. They were shown to be dependent on CMG for replication and movement within the plants, confirming their status as satellite molecules named here as satDNA-II and satDNA-III. When present in coinfection with CMGs, they enhance symptoms and can break high levels of resistance in a cassava landrace. Finally a simple, inexpensive technique is described of archiving, transporting and recovering plant DNA for downstream geminivirus characterisation. / Thesis (PhD)--University of Pretoria, 2007. / Microbiology and Plant Pathology / Unrestricted