Call number: LD2668 .T4 1965 R662 / Master of Science

Call number: LD2668 .T4 1955 K35 / Master of Science

Call number: LD2668 .T4 1966 R161 / Master of Science

A dissertation submitted in partial compliance with the requirements for the Masters Degree in Technology: Homoeopathy, Technikon Natal, 1997. / Most economically important crop plants may become infected with viruses. Several of these virus diseases are limiting factors in agricultural production and have contributed to serious economic and social hardship in many countries, especially in tropical and subtropical regions. Homoeopathic microdoses have been investigated for their role in the control of virus diseases, with good results. However, few of the studies contain statistical analyses. The object of this study was to assess the effect of a homoeopathic preparation of a leaf infected with tobacco mosaic virus (TM Viricum) in the contol of tobacco mosaic virus (TMV). The potencies used were 6CH, 12CH, 30CH and 200CH. iv Trays of 24 tomato seedlings per tray were the subjects of this study. Tomato plants were systemically infected with TMV. Four trays were used per treatment. There was an uninoculated and an inoculated control group. The rest of the test population was divided into two groups. The / M

Call number: LD2668 .T4 1957 H31 / Master of Science

Geminiviruses causes diseases to many staple food and cash crops of great economic importance worldwide. Currently eight species of Begomoviruses belonging to the Geminivirus family exist, of which South African cassava mosaic virus SACMV-[ZA:99] is a member, and is known to cause cassava mosaic disease (CMD). Cassava (Manihot esculenta, Crantz) is considered to be an important food crop consumed in many tropical, sub-tropical and African countries, and is increasingly becoming well-known for its ethanol production on a global a scale. Various strategies to control CMD are currently being implemented, one of which is to elucidate mechanisms involved in host-virus interactions with the aim of identifying defence-related genes involved in the disease process. Many defence genes within the plant kingdom are evolutionary conserved, potentially providing methods of control not only to CMD but to other diseases as well. The research outlined in this thesis aimed to identify networks and pathways involved in disease susceptibility between the model plant host system, Arabidopsis thaliana and cassava T200 upon SACMV-[ZA:99] infection. Conclusions were also drawn from within host comparisons between susceptible cassava T200 and resistant cassava TME3 cultivars in order to explore if similarities, differences or common patterns of expression existed between genes governing resistance and susceptibility. Before transcriptomic profiling studies were carried out, it was important to improve South African cassava mosaic virus (SACMV-[ZA:99]) and African cassava mosaic virus (ACMV-[NG:Ogo:90]) infection efficiencies in recalcitrant crop systems such as cassava. Susceptible cassava cultivars T200, TMS60444, and SM14334 were tested for these purposes following infection with three different Agrobacterium strains (C58C1; AGL1; LBA4404). Results demonstrated that an overall increase in infection efficiency was achieved for each genotype and virus tested, although with varying infectivity levels, suggesting that although an improved method was established, basal levels of susceptibility differed between genotypes and therefore it was not possible to achieve 100% infection efficiencies for agroinfection methods. A 4 x 44k microarray whole genome study was then conducted to identify susceptible host genes involved in the interaction between the model plant system Arabidopsis thaliana and SACMV-[ZA:99]. An infectivity assay was carried out across three time points (14, 24, and 36 dpi), confirming that disease symptoms and virus infectivity levels correlated with an increase in differentially expressed transcripts across time points, with SACMV-[ZA:99] predominantly causing host-gene suppression. Many complex genes and pathways were disrupted and were shown to be involved in categories pertaining to stress and defence responses, phytohormone signalling pathways, cellular transport, metabolism and cell-cycle regulation strongly suggesting an attempt made by SACMV-[ZA:99] to affect homeostasis and antagonize host defence responses. This was the first geminivirus study identifying differentially expressed transcripts across 3 time points. Next generation sequencing (NGS) using the ABI Solid platform was then carried out on SACMV-[ZA:99] – infected susceptible cassava T200 cultivar at 3 time points (12, 32, and 67 dpi), comparing infection responses to mock-inoculated healthy controls. Similarly to the Arabidopsis microarray study, findings from this analysis also revealed a shift from up-regulated to down-regulated genes across time points, once again reflecting virus-specific suppression on host genes suggesting SACMV-[ZA:99] specific alterations were induced in the host, regardless of the host (Arabidopsis and cassava T200) or platform (microarray and NGS) used. Genes identified pertaining particularly to the susceptible cassava T200 - SACMV-[ZA:99] interaction such as the disease resistance protein families (TIR-NBS-LRR), RPP1, RPM1, and NHO1 were showing down-regulation demonstrating that SACMV-[ZA:99] pathogenicity proteins may be causing this suppression leading to inactivation of basal immunity. Comparisons between tolerant cassava TME3 and susceptible T200 showed similarities and differences in responses between the cultivars. Many similarities such as cell wall precursor proteins and glutathione-S-transferases were up-regulated in both cultivars, which may be due to the host attempting to mount appropriate defences. Opposite patterns of expression was observed for genes in categories involved in transcription and phytohormone signalling such as WRKY‘s, NAC, JAZ, and ERF where suppression was evident in susceptible cassava T200, confirming the suppressive nature of SACMV-[ZA:99] to establish a replication-competent environment. Findings in this study contributed to the little that is known about geminivirus disease progression within a previously uncharacterised susceptible host such as cassava.

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iv, 114 leaves : ill. ; 25 cm / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Plant Pathology, Waite Agricultural Research Institute, 1972

Some positive-strand RNA plant viruses possess a transfer RNA-like structure (TLS) at the 3'-terminus of their genomic RNAs. The closest mimicry to tRNA is exhibited by the valylatable TLSs from tymoviruses and furo-like viruses, which are able to interact with key cellular tRNA enzymes: [CTP, ATP]:tRNA nucleotidyltransferase (CCA NTase), valyl-tRNA synthetase (ValRS), and translation elongation factor 1A (eEF1A). In this thesis, I report the discovery of two new roles of the Turnip yellow mosaic tymovirus TLS, in translation enhancement (Chapter 2) and repression of minus strand initiation (Chapter 4). Placement of the 3'-terminal 109 nts of TYMV RNA in a luciferase reporter RNA with a generic 5'-UTR enhanced translation by about 20-fold in cowpea protoplasts. Exhibiting a synergistic relationship with the 5'-cap, the 3'-translation enhancement was largely dependent on the aminoacylatability of the TLS and apparently on eEF1A interaction. In the presence of the 5'-UTR from genomic TYMV RNA, translation of both the overlapping proteins p69 and p206 was strongly dependent on a 5'-cap structure, and was enhanced by the 3'-enhancer. These in vivo results contradict the proposed model in which translation initiation of p206, but not p69, is cap-independent and TLS-dependent (Barends et al. Cell 112(2003):123-9). In vitro experiments with a partially purified preparation of TYMV replicase have investigated the phenomenon of minus strand repression. Interaction of purified eEF1A���GTP specifically with the valylated TLS decreased the template activity for minus strand to near-background levels. eEF1A���GTP acts by making the 3'-CCA minus strand initiation site unavailable to the replicase. The influence of eEF1A in simultaneously enhancing translation and repressing minus strand synthesis can be considered a regulation that ensures robust translation early in the infection and that offers a coordinated transition from translation to replication. Previously shown to be critical for TYMV infectivity, a valylatable TLS was investigated for its role in the replication and infectivity of the bipartite Peanut clump pecluvirus. A valylatable TLS provided a small competitive advantage in protoplasts and whole plants. The advantage was more apparent in protoplasts than in whole plants, and more so in the replication protein-encoding RNA1 than in the trans-replicating RNA2. / Graduation date: 2004