Thesis (MSc (Genetics))--University of Stellenbosch, 2005. / An infectious clone of a viral RNA genome is one that can be used, either as an in vitro transcript or as cDNA, to produce an infection in a susceptible plant. Infectious clones serve as a tool to study viral RNA genomes at a molecular level to gain deeper insight into genome organization, viral gene function, presence of regulatory sequences and gene expression. In the Western Cape (and elsewhere) a new crippling grapevine disease, known as Shiraz disease, is emerging of which the aetiology and pathogenic agents involved are not yet fully understood. Grapevine virus A (GVA), genus Vitivirus, family Flexiviridae, is thought to be the associated with this disease. The aim of this study was to construct a full-length infectious cDNA clone of GVA, which will aid in the molecular study of the viral genome. This clone could ultimately be used to investigate GVA’s involvement in Shiraz disease, which could lead to the unravelling of the aetiology and control of the disease. A full-length clone of GVA, named GVA-IC2/T7-2972-3, was constructed in several steps using restriction digestion/ligation and primer overlap extension PCR. Grapevine virus A cDNA fragments were obtained from GVAinfected Nicotiana benthamiana and Vitis vinifera plants using three different techniques, of which the Rapid direct-one-tube RT-PCR was most successful. A 5’ T7 promoter and a 3’ poly-A tail were incorporated and the full-length clone was cloned into pBluescript II SK (+). Full-length sequencing of the clone, revealed two significant frameshift mutations. The first mutation was a single base pair insertion (one G) in a slippery site of 6 G’s at position 1380 – 1385 in open reading frame one (ORF 1) of the viral genome. This mutation was corrected by PCR-based site-directed mutagenesis, which resulted in pSK-GVA-mutagen-3 and pSK-GVA-mutagen-4. The second mutation was a single base pair deletion (one G) at position 6959 in ORF4, which coded for the coat protein (CP). Several techniques were attempted to correct this mutation, but none were successful. Even though the second mutation could not be corrected, in vitro transcriptions were performed on three clones followed by subsequent infections of N. benthamiana plants. The three clones included pSK-GVA-mutagen-3, pSKGVA- mutagen-4 (both hosting the mutation at position 6959) and GVA-IC2/T7-2972-3 (hosting both mutations). At 21 days post-inoculation no significant visual symptoms were observed in plants infected with in vitro RNA or in plants infected with wild type GVA. Rapid direct-one-tube RT-PCR results revealed the presence of viral RNA in infected leaves and apical leaves of infected plants, and provided preliminary evidence that the mutated clones were still capable of systemic infection and viral movement. These results are still inconclusive, and several post-infection studies will have to be performed to confirm these findings. Koch's postulates will also have to be proved in order to confirm the infectious nature of the clones. The effect of the two mutations in the constructed clones will be investigated further and post-infection analysis performed to deduce whether the viral progeny are devoid of the mutations. Three full-length GVA cDNA clones (hosting mutations) seemingly capable of systemic infection in N. benthamiana plants were constructed in this study and have laid the foundation for molecular and mutational analysis of the GVA genome. This could lead to the study of pathogen-host interactions in order to unravel the aetiology of Shiraz disease in the future.

Thesis (MScAgric)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: Leaf blackening, a postharvest disorder which is characterized by a dark brown to black discoloration, is found in most commercially important Protea cut flower species and cultivars. As this disorder is known to increase with storage time, it is a major concern to the South African industry as the use of sea freight is increasingly preferred due to lower transport costs and a more favourable carbon footprint. The cause of leaf blackening has been strongly linked to a carbohydrate stress exerted by the large inflorescence, thus requiring the utilization of sugar bound polyphenols in the foliage, which when removed, can oxidize enzymatically or non-enzymatically. A study where harvesting was done throughout the season as well as on selected days at 08:00, 12:00, 15:00 and 17:00, concluded that leaf blackening incidences in Protea cv. Sylvia stems varies significantly throughout the season, between years and even with the harvest time of day. Leaf blackening incidences increased from October onwards and remained high until February, before decreasing to acceptably lower levels towards March to May. Carbohydrate- and phenolic content together with water status of leaves at harvest was not able to accurately predict incidence of the associated leaf blackening. However, irrespective of the season of harvesting, leaf blackening was significantly lower when stems were harvested later in the day than compared to stems harvested in the morning. Low sucrose and high water content at these harvest times was positively correlated to high incidences of leaf blackening. In a next study where uptake dynamics of glucose pulsing was investigated, Protea cv. Sylvia was harvested at different times throughout the day, dehydrated to various levels and pulsed with an increasing range of glucose concentrations. Pulsing solution uptake per stem was found to be highly influenced by these factors, as dehydration of stems and a harvest time later during the day both decreased stem water potential, which then increased pulse-solution uptake within a certain time period. The daily harvest time influenced transpiration, whilst pulse-solution uptake decreased with an increase in glucose pulse concentration. When stems were pulsed pre-storage with an increasing range of glucose concentrations, not only did pulses of between 4.7 – 13.7% glucose significantly delayed the incidence of leaf blackening, but it also maintained a positive water balance longer in stems during vase life. Ethanol or acetaldehyde vapour did not provide a viable alternative for reducing leaf blackening incidence in Protea cv. Sylvia, although a synergistic effect was found when ethanol vapour or pulsing was used in combination with glucose. A commercial verification trial disclosed that Protea magnifica and Protea ‘Pink Ice’ reacted more beneficial to ethanol vapour than was observed in ‘Sylvia’. This study confirms that carbohydrate availability within the Protea cut stem remains a key factor in the control of leaf blackening. Factors which assist in maintaining high internal carbohydrate levels, such as enhanced glucose pulse uptake or effective vase solution utilization will contribute to providing an optimum control of leaf blackening during vase life following long-term cold storage. / AFRIKAANSE OPSOMMING: Loofblaarverbruining is ‘n na-oes defek wat gekarakteriseer word deur ‘n donker bruin na swart verkleuring wat voorkom in meeste kommersieël belangrike Protea snyblom spesies en kultivars. Hierdie defek is bekend daarvoor dat dit toeneem met stoortyd, dus is dit ‘n groot kommer vir die Suid-Afrikaanse industrie, met toenemende gebruik van seevrag as vervoer keuse wat laer vervoer kostes en meer gunstige ‘koolstof voetspoor’ bevoordeel. Die oorsaak van loofblaarverbruining word sterk gekoppel aan ‘n koolhidraat stres wat uitgeoefen word deur die groot bloeiwyse op die loofblare, waar suiker-gebonde polifenoliese verbindings ensiematies of nieensiematies geoksideer word met die verwydering van die suiker verbinding. 'n Studie waar geoes was regdeur die seisoen, sowel as op geselekteerde dae om 08:00, 12:00, 15:00 en 17:00, het bevind dat die voorkoms van loofblaarverbruining in stele van Protea kv. Sylvia aansienlik geskil regdeur die seisoen, tussen jare en selfs met die oes tyd gedurende die dag. Die voorkoms van loofblaarverbruining het toegeneem vanaf Oktober en het hoog gebly tot en met Februarie, voordat dit gedaal het tot aanvaarbare laer vlakke teen Maart, tot en met Mei. Koolhidraat-en fenoliese inhoud sowel as die water status van die blare by oes was onsuksesvol om die voorkoms van die gepaardgaande loofblaarverbruining akkuraat te voorspel. Loofblaarverbruining was egter aansienlik laer as stele geoes later in die dag teenoor stele geoes in die oggend, ongeag die seisoen van oes. Lae sukrose en 'n hoë water inhoud geassosieer met hierdie oes-tye was positief gekorreleerd met ‘n hoë voorkoms van loofblaarverbruining. In 'n volgende studie waar die opname dinamika van glukose pulsing ondersoek was, is Protea kv. Sylvia stele geoes op verskillende tye dwarsdeur die dag, gedehidreer tot verskillende vlakke en met 'n toenemende reeks van glukose konsentrasies gepuls. Pulsoplossing opname per steel is sterk beïnvloed deur hierdie faktore, aangesien dehidrasie van die stele asook stele geoes later gedurende die dag die afname van steel waterpotensiaal veroorsaak het, terwyl die puls-oplossing opname versnel het binne ‘n bepaalde tyd. Die tyd van oes beïnvloed ook transpirasie, terwyl vaas oplossing opname afgeneem met 'n toename in glukose puls konsentrasie. Wanneer ‘Sylvia’ stele gepuls was voor stoor met 'n reeks van toenemende glukose konsentrasies, het nie net die puls van tussen 4.7 – 13.7% glukose aansienlik die voorkoms van loofblaarverbruining vertraag nie, maar dit het ook ‘n positiewe water balans langer in stele gedurende die vaas lewe behou. Nie etanol of asetaldehied dampe is bevind as geskikte alternatief vir glukose pulsing om die voorkoms van loofblaarverbruining in Protea kv. Sylvia te verlaag nie, alhoewel ‘n sinergistiese effek waargeneem was wanneer etanol in kombinasie met glukose gebruik was. ‘n Kommersieële bevestigingstoetsing het bevind dat Protea magnifica en ‘Pink Ice’ meer voordeel uit ‘n ethanoldamp behandeling kon trek teenoor ‘Sylvia’. Hierdie studie het bevestig die belangrikheid van koolhidraat beskikbaarheid in die Protea snyblom, vir beheer van loofblaarverbruining. Faktore wat die handhawing van hoë interne koolhidrate vlakke, soos bevorderde glukose puls opname of effektiewe vaas oplossing benutting sal bydra tot ‘n optimal beheer van loofblaarverbruining tydens vaas lewe na langtermyn koueopberging. / National Research Fund (NRF) for their financial support in 2009; Protea Producers of South Africa (PPSA) and Productschap Tuinbouw (PT) as well as the Frank Batchelor Will Trust Grant for the financial support.

False codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is considered the most important indigenous pest of citrus in southern Africa. Major concerns such as progressive insecticidal resistance, the negative impact of insecticides on the environment, as well as the influence of consumers opposed to chemical residues on fruit, created opportunities for biological control methods such as Sterile Insect Technology (SIT). This technology is now established in the Western and Eastern Cape provinces of South Africa as an effective, sustainable alternative to conventional FCM control methods. Due to the prevalence of the pest in all citrus producing areas of South Africa, potential for SIT to expand is enormous. Success of an SIT programme is highly dependent on efficient application of the technology to achieve its objectives in a timeous manner. The aim of this study was to advance the application of SIT for control of FCM on citrus in South Africa, by investigating the effect of certain critical stages in the process. The effect of long-distance transportation on fitness of irradiated FCM was determined, showing reduced performance with cold-immobilized transport. A significant decrease in flight ability and longevity of irradiated FCM was found, although critically, realized fecundity was not affected. The effect of two different insecticides in the pyrethroid and organophosphate chemical groups were investigated for their residual effect on mortality of released irradiated FCM, to determine if these pest control programmes could be integrated. Both chlorpyrifos and tau-fluvalinate were effective in killing irradiated FCM for a number of days after application, after which degradation of the active ingredient rendered it harmless. This effect was found to be similar for irradiated and non-irradiated males, consequently ratios of sterile : wild male FCM should be retained regardless of whether sprays are applied or not. The modes for release of sterile FCM in an SIT programme were investigated. Efficacy of ground and aerial release platforms were tested by evaluating the recovery of released irradiated male FCM in these orchards. More irradiated FCM were recovered in orchards released from the ground compared to air. However, an economic analysis of both methods shows application of irradiated insects over a large geographical area is more cost-effective by air. Depending on the terrain and size of the target area, a combination of both methods is ideal for application of SIT for control of FCM in citrus. Development of application technology for advance of the programme is discussed and recommendations for future research and development are offered.

Made available in DSpace on 2014-06-11T19:34:59Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-05-29Bitstream added on 2014-06-13T19:04:59Z : No. of bitstreams: 1 prado_ep_dr_botfca.pdf: 729371 bytes, checksum: 43b42061e06f38fc114ff3ebf2484262 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Com a finalidade de comparar diferentes tecnologias de aplicação em pulverizações terrestre no controle da ferrugem asiática da soja (FAS), o trabalho objetivou construir um dispositivo abridor do dossel da soja e comparar diferentes tecnologias de aplicação de fungicida sobre os depósitos, controle da FAS e produtividade da soja. A confecção do dispositivo Canopy opener (CO) foi baseada no protótipo desenvolvido por pesquisadores do Departamento de Agricultura dos Estados Unidos da América (USDA). Em laboratório verificou-se a eficiência da pulverização com CO e a pulverização convencional (Experimento 1). Os ensaios de campo foram conduzidos na área experimental da FCA/UNESP - Campus de Botucatu nos anos agrícola 2009/10 (cultivar FTS Campo Mourão RR) e repetidos no ano agrícola 2010/11 (cultivar BRS Valiosa RR). No campo foram comparadas as pulverizações realizadas com auxílio da assistência de ar (AA) junto à barra de pulverização, pulverização feita com CO e a pulverização associada com AA e o disposivo CO (Experimento 2). Estudou-se o efeito dos volumes de aplicação de 110, 160 e 210 L ha-1 com e sem o uso da AA (Experimento 4) e os volumes de 60, 110 e 160 L ha-1 na presença e ausência do surfatante siliconado (Experimento 4). Em todos os ensaios foram mensurados os depósitos da pulverização em diferentes partes da planta. Para os ensaios a campo, além da deposição, foram avaliadas a severidade da FAS, mensurado o peso de mil grãos (PMG) e a produtividade. O dispositivo CO construído, além de apresentar viabilidade para realização dos estudos, proporcionou maiores valores de depósitos nas folhas da parte inferior, e menores nas folhas da parte superior das plantas de soja comparada com... / With purpose to compare different ground spray technology of application on Asian soybean rust (ASR) control, this research aimed to build an equipment that facility the spray penetration into soybean canopy and compare different fungicide application technologies on spray deposition, ASR control, and soybean yield. The equipment called canopy opener (CO) was built based in a prototype developed by United States Department of Agriculture researchers. In laboratory was checked the spraying efficiency with CO and compared to conventional spraying (Experiment 1). The field trials were carried out at experimental farm located in Sao Paulo State University (UNESP), Botucatu Campus - Brazil in 2009/10 agricultural season (FTS Campo Mourão RR variety) and replicated in 2010/11 agricultural season (BRS Valiosa RR variety). In field trials were compared spraying using sleeve boom sprayer with air-assistance (AA), spraying using CO on spray boom and spraying with association between CO and AA (Experiment 2). The effects of different spray rates (110, 200 and 160 L ha-1) with and without AA (Experiment 4) and spray rates of 60, 110 and 160 L ha-1 with and without silicone surfactant (Experiment 4) were studied. In all of trials were measured the spray deposits on different canopy portion. In addition to the spray deposition were assessed the ASR severity, thousand seed weight (TSW) and the productivity in yield trials. The CO device showed to be viable for this research, as well as provided greater deposits values on canopy leaves of the bottom part of the plants and lower deposit levels on top canopy leaves when compared with conventional spraying in laboratory. The CO showed equivalent results to the air-assisted spraying on spray deposition, soybean rust control represented by the area under disease progress... (Complete abstract click electronic access below)

Orientador: Carlos Gilberto Raetano / Banca: Vilson Luis Kunz / Banca: Ulisses Rocha Antuniassi / Banca: Marco Antonio Gandolfo / Banca: Edson Luiz Furtado / Resumo: Com a finalidade de comparar diferentes tecnologias de aplicação em pulverizações terrestre no controle da ferrugem asiática da soja (FAS), o trabalho objetivou construir um dispositivo abridor do dossel da soja e comparar diferentes tecnologias de aplicação de fungicida sobre os depósitos, controle da FAS e produtividade da soja. A confecção do dispositivo Canopy opener (CO) foi baseada no protótipo desenvolvido por pesquisadores do Departamento de Agricultura dos Estados Unidos da América (USDA). Em laboratório verificou-se a eficiência da pulverização com CO e a pulverização convencional (Experimento 1). Os ensaios de campo foram conduzidos na área experimental da FCA/UNESP - Campus de Botucatu nos anos agrícola 2009/10 (cultivar FTS Campo Mourão RR) e repetidos no ano agrícola 2010/11 (cultivar BRS Valiosa RR). No campo foram comparadas as pulverizações realizadas com auxílio da assistência de ar (AA) junto à barra de pulverização, pulverização feita com CO e a pulverização associada com AA e o disposivo CO (Experimento 2). Estudou-se o efeito dos volumes de aplicação de 110, 160 e 210 L ha-1 com e sem o uso da AA (Experimento 4) e os volumes de 60, 110 e 160 L ha-1 na presença e ausência do surfatante siliconado (Experimento 4). Em todos os ensaios foram mensurados os depósitos da pulverização em diferentes partes da planta. Para os ensaios a campo, além da deposição, foram avaliadas a severidade da FAS, mensurado o peso de mil grãos (PMG) e a produtividade. O dispositivo CO construído, além de apresentar viabilidade para realização dos estudos, proporcionou maiores valores de depósitos nas folhas da parte inferior, e menores nas folhas da parte superior das plantas de soja comparada com... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: With purpose to compare different ground spray technology of application on Asian soybean rust (ASR) control, this research aimed to build an equipment that facility the spray penetration into soybean canopy and compare different fungicide application technologies on spray deposition, ASR control, and soybean yield. The equipment called canopy opener (CO) was built based in a prototype developed by United States Department of Agriculture researchers. In laboratory was checked the spraying efficiency with CO and compared to conventional spraying (Experiment 1). The field trials were carried out at experimental farm located in Sao Paulo State University (UNESP), Botucatu Campus - Brazil in 2009/10 agricultural season (FTS Campo Mourão RR variety) and replicated in 2010/11 agricultural season (BRS Valiosa RR variety). In field trials were compared spraying using sleeve boom sprayer with air-assistance (AA), spraying using CO on spray boom and spraying with association between CO and AA (Experiment 2). The effects of different spray rates (110, 200 and 160 L ha-1) with and without AA (Experiment 4) and spray rates of 60, 110 and 160 L ha-1 with and without silicone surfactant (Experiment 4) were studied. In all of trials were measured the spray deposits on different canopy portion. In addition to the spray deposition were assessed the ASR severity, thousand seed weight (TSW) and the productivity in yield trials. The CO device showed to be viable for this research, as well as provided greater deposits values on canopy leaves of the bottom part of the plants and lower deposit levels on top canopy leaves when compared with conventional spraying in laboratory. The CO showed equivalent results to the air-assisted spraying on spray deposition, soybean rust control represented by the area under disease progress... (Complete abstract click electronic access below) / Doutor

M.Sc. / Plant disease can be naturally suppressed by plant growth promoting rhizobacteria and endophytic / endorhizosphere bacteria. Apart from direct antagonism against pathogenic organisms, these plant growth promoting bacteria and endophytes can induce a form of systemic resistance (ISR) in plants. The main bacterial inducing component has been suggested to be the outer membrane lipopolysaccharides (LPS), found in the cell walls of Gramnegative bacteria. Burkholderia cepacia (Pseudomonas cepacia) is a bacterial endophyte that has potential as a biocontrol agent. Although a few studies have indicated that LPS from, certain Pseudorrionads has a protective effect in plants against disease, a controlled investigation has not been attempted previously with a purified preparation of LPS. LPS was isolated from the bacterial cell wall, prepared and characterized by denaturing electrophoresis. Characterization of the LPS also included the determination of 2-keto-3-deoxyoctonate, carbohydrate —, as well as the protein content. The purified LPS was found to possess activity as an elicitor of plant defence responses in tobacco where the induction of pathogenesisrelated (PR) proteins were investigated and electrophoretically analysed. An optimum LPS concentration range of 50-150 14/m1 was determined by studying cell death using the Evans blue procedure. Time and concentration ranges for LPS induced responses were established in cell suspensions, leaf discs, whole leaves and whole plants. It was determined that the PR-protein response could be optimally induced after four days following elicitation with 100 fag/ml LPS. Systemic induction of resistance was tested by treatment of the lower leaves and following the response in the upper leaves; as well as bacterial inoculation of the plant roots followed by PR-protein extraction of the leaves. Treatment of tobacco plants with LPS protected the plants against subsequent infection by the pathogen Phytophthora nicotianae, thereby suggesting a role for LPS as activators of systemic acquired resistance (SAR). It can be concluded from this study that the lipopolysaccharides from Burkholderia cepacia, that were used in this study, are effective local as well as systemic inducers of the defense PR-proteins in Nicotianae tabacum cv Samsun NN. The fact that protection is associated with PR-protein induction distinguishes it from the protection induced by rhizobacteria.

A dissertation submitted in partial compliance with the requirements for the Masters Degree in technology: Homoeopathy, Technikon Natal, 1999. / The most common and difficult insect to control in the greenhouse is the whitefly. Sweetpotato whitefly (Bemisia tabaci Genn.) was the species of whitefly selected to be investigated in this trial. Sweetpotato whitefly is a major pest on a variety of plants, ranging from ornamental flowers to agricultural crops such as tomato and cotton, resulting in important financial losses. Effective control options of sweetpotato whitefly are limited. / M

M.Sc. / in the current study the effect of the addition of methyl Jasmonate (MeJA), chitosan, a cell wall elicitor prepared from Phytophthora nicotlanae to tobacco cells and the subsequent defense responses elicited in these cells were Investigated. The defense responses investigated can be divided into three categories according to the time scale whereby resistance responses in plant cells are induced: early events which included the analysis of lipid peroxidation, the induction of lipoxygenase (L0)0 enzyme activity as well as the changes in phosphoprotein profiles; intermediate to later responses which included investigations of peroxidase (POD) activity, lignin content, phytoalexin content and phenolic content and also late responses which included studies of pathogenesis-related proteins (PR) and 13-1,3-giucanase activity. An approach also followed in this study was the addition of MeJA to tobacco cells for 24 h followed by the addition of either the cell wall elicitor or chitosan as a secondary elicitors, to investigate possible preconditioning or sensitisation by MeJA. Results obtained in this study revealed the time and concentration dependent accumulation of phytoalexins (secondary metabolites) when MeJA was added to tobacco cells and an optimal concentration of MeJA to use in further studies was determined as 1 mM. MeJA was the most effective inducer of lipid peroxidation (22 fold induction), a response observable after 2 h of exposure to MeJA. Conditioning with MeJA, followed by both chitosan (19 fold induction) and elicitor (25 fold induction) led to an earlier accumulation as well as significant increases in the levels of malondialdehyde, the product of lipid peroxidation. LOX enzyme activity was significantly increased by the addition of MeJA (6 fold Induction), chitosan (4 fold induction) and elicitor (3.8 fold induction). Conditioning with .MeJA, followed by both chitosan (3.3 fold induction) and elicitor (3.9 fold Induction) also led to noteworthy increases in enzyme activity. Analysis of the phosphoprotein profiles do not reveal the accumulation of phosphorylated proteins when MeJA was added to cells and very little accumulation of such proteins when chitosan was added. Phosphorylated proteins could be observed in cells treated with elicitor and In the cases where conditioning with MeJA, followed by secondary elicitation with either chitosan or elicitor, was studied, the differential induction of phosphorylated cellular proteins could also be observed. No significant induction of POD activity could be observed under any of the conditions, except for a possible slight increase in POD activity starting at 16 - 24 h after the elicitor had been added and a more definite increase after 24 h which was sustained up to 48 h after the addition of MeJA. PAGE of peroxidase, followed by activity staining revealed the presence of a slow migrating anionic peroxidase as well as a fast migrating peroxidase. Conditioning with MeJA, followed by secondary elicitation with both chitosan and elicitor revealed enhanced POD activity as well increased induction of a fast migrating anionic peroxidase on PAGE gels. MeJA was a more effective inducer of elevated levels of lignin content than the elicitor or chitosan and the addition of MeJA to tobacco cells led to a 2.2 fold increase in the lignin content, a response observed after 24 h and sustained up to 48 h. Chitosan as secondary elicitor did not lead to any increase in lignin content, but the cell wall elicitor as secondary agent significantly increased the lignin content after 40 - 48 h. Analysis of phenolic content did not show any significant increases In the total soluble phenolics when the agents were used on their own and only the phenolic content of the MeJA-conditioned cells, followed by the addition of chitosan showed a slight increase. In this case, the HPLC analysis of the phenolics also revealed a shift In the profiles for phenolics. SDS-PAGE of PR proteins revealed the induction of constitutive as well as new proteins when MeJA and elicitor, but not chitosan were used as elicitation agents. However, In the MeJA-pretreated cells addition of both chitosan and elicitor led to increased accumulation of PR proteins with molecular masses ranging from 6 - 70 kDa. Results from the i3-1,3-glucanase activity assay indicate a strong induction (4-5 fold) when MeJA and elicitor (4 fold), but not when chitosan was added to cells. Conditioning effects were revealed when both chitosan (3 fold induction) and elicitor (2.5 fold induction) were used as secondary elicitors. The increases in intensities of bands with molecular masses ranging from 31- 35 kDa observed on SOS-PAGE gels where chitosan and elicitor were added as secondary agents corresponded in a time dependent manner with the increased levels obtained in thep-1,3-glucanase activity assay.

Please read the abstract in the section 00front of this document. Also note that an abstract is provided for each chapter as well / Thesis (DPhil (Entomology))--University of Pretoria, 2005. / Zoology and Entomology / unrestricted

During the initiation and execution of a rootstock breeding programme to overcome the financially crippling disease, Phytophthora root rot of avocado, various constraints have been identified for both the breeding as well as the screening aspect of the programme. A review of the literature revealed a complex host-pathogen interaction that should be taken into account in the recombination and screening of genetic material. With the detection of beneficial genotypes being the crux of a breeding programme, this dissertation was focused on the screening of rootstock material for tolerance to Phytophthora cinnamomi. Screening should be scientific but at the same time also be time and cost effective. Specific attention was given to (i) the correct medium for screening mass numbers of seedlings, (ii) fast and effective cloning of single selections, and (iii) evaluation of clonal material for tolerance to P. cinnamomi. Soil as a screening medium was compared with three inert hydroponic media as well as one aeroponic system. Only soil was found to be ineffective due to its properties. The other media tested, namely, sand, vermiculite, water and the aeroponic system were equal in performance. The medium to be used will depend on the preference of the breeder as each medium has its own pro's and con's. It was, however, found that the evaluation criterion to be applied depends on the medium that is used. With regard to cloning of single selections, a definite difference with regard to the cloning ability of the different selections was found. An inability to be etiolated was displayed by some of the selections and these could thus not be vegetatively propagated and were not further tested. One of the tolerance mechanisms in the standard cultivar Duke 7, is root regeneration. It was thus expected that this characteristic cloning would give an indication of the rootstock's ability to tolerate P. cinnamomi. This could not be confirmed, but most of the selections did, however, perform better than Duke 7. Comparison of feeder root percentage in non-inoculated and inoculated treatments was not sufficient for facilitating the final selection of candidate rootstocks from a large number of potential clonal selections. Four selections were made, based on the hypothesis that a larger root system will be a better forager and thus enhance the horticultural aspects of the rootstock-scion combination. Valuable information was obtained with regard to various mediums and criteria to be used during mass screening and final screening of clonal selections. This knowledge must be taken into account in the planning of future breeding projects. During this project a total of 38 984 seedlings were screened and four selections were made. For both the nursery and the producer, knowledge of the clonal ability of a potential new rootstock is important from a financial point of view. / Dissertation (MSc (Horticultural Science))--University of Pretoria, 2006. / Plant Production and Soil Science / unrestricted