El presente trabajo de investigación es el resultado del trabajo continuo y perseverante en el estudio de la producción de uvas de mesa en la región de Ica. Para ello, se focalizó el estudio en las mypes, ya que uno de los objetivos de este trabajo es contribuir con el conocimiento adquirido para organizar estas empresas. Por esta razón, se utilizó la gestión por procesos para ordenar la empresa desde el punto de vista del valor agregado que le podemos ofrecer al cliente. Este trabajo está enfocado en el proceso de mantenimiento en producción, cuyo principal objetivo es mitigar todos aquellos factores que impidan el crecimiento de la uva de mesa. Es decir, tomando como base el mantenimiento autónomo, se proponen actividades de mantenimiento para tener en óptimas condiciones el terreno de cultivo, el sistema de irrigación y la estructura del parral. Esto permitirá que la uva pueda tener las características físicas necesarias para la venta a grandes acopiadores. Cabe resaltar que este es el medio por el cual los agricultores pueden adquirir mayor margen de ganancia. Finalmente, todas las propuestas en esta tesis han sido realizadas para las personas que administran estas mypes. En su mayoría, son personas con estudios secundarios y no cuentan con especializaciones para poder dirigir sus empresas. / The present research work is the result of continuous and persevering work in the study of grape production in the Ica region. For this, the study was focused on the mypes, since one of the objectives of this work is to contribute with the knowledge acquired to organize these companies. For this reason, process management was used to order the company from the point of view of the added value that we can offer to the client. This work is focused on the maintenance process in production, whose main objective is to mitigate all those factors that impede the growth of grapes. Based on autonomous maintenance, related maintenance activities were proposed to keep the cultivation land, the irrigation system and the structure of the vine in optimal conditions. This will allow the grape to have the physical characteristics necessary for sale to large collectors. It should be noted that this is the means by which farmers can acquire a higher profit margin. Finally, all the proposals in this thesis have been made for the people who administer these mypes. For the most part, they are people with secondary education and do not have specializations to be able to run their companies. / Tesis

Vermicompost transforms agricultural waste into a high quality soil amendment, though market acceptance remains in its infancy. This study examines how growers’ willingness to pay for vermicompost is affected by grower’s crop, region, income per acre, knowledge level of vermicompost and compost, previous use of compost, and the willingness to pay for compost. The survey results pulled together 223 responses from California growers. It was discovered growers’ had less knowledge of vermicompost than compost but were willing to pay more for vermicompost. There was statistical difference amongst the responses. It was shown the market value per ton of vermicompost lies between $20 and $30; and feasibility of a potential vermicompost facility depends on transportation costs of the finished product. A vermicompost company should focus on North Coast grape growers.

Drosophila suzukii (Matsumura) (Diptera: Drosophila) is an invasive frugivore and has become a significant pest of small fruit, cherry and grape throughout the United States. It may be possible to determine if a Virginia vineyard is at risk of D. suzukii infestation by analyzing the biotic and abiotic factors around each vineyard. This pest is known to utilize a wide range of cultivated and wild host plants. A host plant survey was conducted at four vineyards in the Piedmont Region of Virginia to identify hosts used by D. suzukii around vineyards. The seasonal availability of host plants and adult emergence from them were tracked. Six host plant species of D. suzukii were identified, some available season-long. Monitoring D. suzukii in cultivated crops is crucial for the timing of spray applications. Homemade and commercially available baits and traps were deployed in two vineyards to determine the efficacy and selectivity towards D. suzukii. The homemade and commercially available baits that contained red wine caught the most D. suzukii, but none were exclusively attractive to D. suzukii. Wine grape susceptibility was assessed in laboratory choice and no-choice ovipositional bioassays. Ovipositional susceptibility was determined by measuring the physiological and morphological parameters using six wine grape varieties. More eggs were laid in grapes as penetration force decreased. Penetration force and not skin thickness was the limiting factor for oviposition. Survivorship of eggs laid in intact grapes was analyzed and survivorship to adulthood was dependent upon variety and survivorship usually exceeded 9% survival seen in previous studies. Larval developmental parameters of D. suzukii were affected by grape variety and the density of Z. indianus. D. suzukii mortality was increased in most cases when in competition with Z. indianus, but was less pronounced when reared in Viognier grapes. My Z. indianus oviposition study demonstrated that they will follow injury created by D. suzukii, and then the Z. indianus larvae may outcompete D. suzukii within the berries. These studies greatly improved our understanding of D. suzukii biology and ecology in Virginia vineyards. / Ph. D. / Drosophila suzukii (Matsumura) (Diptera: Drosophila), henceforth referred to as spotted wing drosophila (SWD) is an invasive pest of small fruits, cherries and grapes grown throughout the United States. This pest has a wide host range including cultivated and wild host plants. A host plant survey was conducted at four vineyards in the Piedmont Region of Virginia to identify those used by SWD. The seasonal availability of host plants and adult emergence from their fruit were tracked throughout the growing season. Six host plant species were found and these host plants were available to SWD throughout the season. Monitoring SWD in cultivated crops is crucial for the timing of spray applications. Homemade and commercially available baits and traps were deployed in two vineyards to determine the efficacy and selectivity towards SWD in the vineyard. Baits containing red wine, whether homemade or commercially available caught the most SWD, but none were exclusively attractive to SWD. Wine grape susceptibility to SWD oviposition was assessed in laboratory no-choice and choice ovipositional bioassays using six wine grape varieties; physiological and morphological parameters were considered. More eggs were laid in grapes as penetration force decreased. Penetration force and not skin thickness was the limiting factor for oviposition. Survivorship of eggs laid in intact grapes was analyzed and up to 50% of the eggs laid in larger grapes survived to adulthood. Larval interactions between SWD and Z. indianus, African fig fly (AFF), were also analyzed based on competition intensity and grape variety in which they were reared. SWD mortality, developmental parameters and pupal volume were impacted when in competition with AFF. SWD mortality was less pronounced, even when in competition with AFF, when reared in Viognier grapes compared to any other grape variety tested. My Z. indianus oviposition study demonstrated that they will flow injury created by D. suzukii and lay oviposit eggs into those wounds. These studies greatly improved our understanding of SWD biology and ecology in Virginia vineyards.

Nitrogen is requisite for yeast growth and metabolism during wine fermentation, and contributes to wine flavor development. Understanding how yeast assimilable nitrogen (YAN) impacts wine chemistry and sensory properties helps inform nitrogen management decisions in both the vineyard and winery. This study used Virginia-grown Chardonel, obtained from a multi-year viticulture study evaluating the impact of nitrogen fertilizer on grapevines. The objective of this work is to determine if vineyard treatments resulted in sufficient juice YAN profile differences to impart chemical and/or sensory differences in resulting wines. Grapes were combined from each field replication of six vineyard nitrogen treatments and pressed to generate six juice lots. Each juice was divided in two for triplicate fermentation using one of two Saccharomyces cerevisiae x bayanus yeast strains: EC1118 or QA23 to investigate the interactive effects of YAN and yeast strain. Juice and wine samples were frozen until chemical and UPLC-PDA amino acid analysis. Fermentation replications of each wine were combined, bottled, and stored at 4ºC for four months until evaluation via sensory sorting task with untrained panelists (n=63). Juice YAN concentrations ranged from 157-201mg N/L, a small variation in terms of commercial winemaking. Chemical differences between wines produced with the same juice but different yeast strains were larger in wines that had lower juice YAN concentrations. DISTATIS analysis of sorting results showed differences but no pattern in sensory perceptions of wines based on nitrogen profile or yeast strain. This range of nitrogen profiles shows limited impact of YAN profile on final wine quality. / Master of Science in Life Sciences / Nitrogen is an essential nutrient in grape growing and winemaking. Grapevines use nitrogen as a nutrient to survive and produce grapes for making wine. In winemaking, yeast turn the sugar and nitrogen in grape juice into alcohol via fermentation, using nitrogen to grow and produce other compounds that contribute to the final flavor of the wine. Understanding how nitrogen qualities and quantities in grape juice impact final wine quality is important so vineyard managers can apply nitrogen fertilizer to meet their needs. This thesis evaluates how different nitrogen quantities and qualities in the initial juice and during white wine fermentation impact the chemical and sensory properties of a final wine product. A Chardonel vineyard is being maintained for a multi-year study where the vineyard is divided for grapes in different areas of the plot to receive one of six different nitrogen fertilizer treatments. Grapes were harvested from each treatment and pressed into juice. Each lot of juice was then split in two for fermentation by one of two yeast strains for a total of 12 wines. Wines were bottled and stored for four months until sensory evaluation. Samples of juice and wine were collected and frozen before chemical analysis for nitrogen and other standard wine industry parameters. For sensory analysis, consumers were asked to taste each wine then sort them into groups based on how similar the wines were perceived to be. Chemical differences were found between wines, but they did not translate into sensory differences. Vineyard managers and winemakers can be aware that in Virginia-grown Chardonel, with modest differences in nitrogen from the vineyard treatments, differences in juice and wine chemical parameters did not impact the final wine product.

Thesis (MScAgric)--University of Stellenbosch, 2004. / ENGLISH ABSTRACT: Soilborne diseases of grapevines represent a complex problem with limited information available, both locally and internationally. Previous research in South Africa indicated that Phytophthora and Pythium spp. were the most widespread and devastating pathogens in grapevine nurseries and vineyards in the Western Cape province. The local grapevine industry is currently expanding; new cultivars, methods and agricultural chemicals are being used which can affect soilborne pathogens. It has therefore become necessary to reassess the status of soilborne pathogens in nurseries, since information in this regard is crucial for the development of disease management practices for the expanding local grapevine industry. Soilborne fungal genera associated with roots and crowns of declining nursery grapevines were assessed in surveys conducted at three different grapevine nurseries in the Western Cape province. Cylindrocarpon, Fusarium, Pythium, and Rhizoctonia spp. were consistently isolated from roots and crowns of declining nursery grapevines. Cylindrocladiella spp. and Phytophthora cinnamomi were infrequently isolated from diseased roots, crowns and soil whereas Pythium spp. were abundant in most of the soils. Results suggest that the status of soilborne fungal pathogens in grapevine nurseries in the Western Cape province has changed over the last 30 years. The DNA phylogeny and pathogenicity of the isolates of Cylindrocladiella were determined. Four species of Cylindrocladiella occur on grapevines in South Africa, namely C. lageniformis, C. parva, C. peruviana, as well as a new species, described in this study as C. viticola, which forms part of the C. infestans species complex. Pathogenicity trials were inconclusive. Ten Fusarium spp. were isolated from roots and crowns of declining nursery grapevines, namely F. acuminatum, F. anthophilum, F. chlamydosporum, F. equiseti, F. nygamai, F. oxysporum, F. proliferatum, F. scirpi, F. semitectum and F. solani. The dominant species was F. oxysporum, followed by F. proliferatum and F. solani. In pathogenicity trials F. oxysporum and F. solani significantly reduced root volume, root dry mass, length of new shoots, stem diameter and number of leaves, but increased the percentage of chlorotic leaves and root rot severity. Fusarium proliferatum also caused a significant reduction in new shoot growth, number of leaves and increased root rot severity compared to the controls. Fusarium so/ani seems to be more virulent than F. oxysporum, followed by F. pro/iferatum. This is the first report of F. oxysporum, F. pro/iferatum and F. so/ani as pathogens of grapevines in South Africa, and the first report of F. proliferatum as a pathogen of grapevines in the world. Phytophthora cinnamomi was isolated at low frequencies from declined grapevines, although present in the rhizosphere soil. It is possible that the extensive use of downy mildew chemicals in grapevine nurseries may protect grapevines from infection by P. cinnamomi. The effect of chemicals used to combat downy mildew on Phytophthora root rot of nursery grapevines was evaluated in a glasshouse. There was very little discernable effect of the chemicals tested relative to the control plants for the parameters measured and it was concluded that the inoculation technique needed refinement. However, plants treated with phosphorous acid tended to be taller and have more leaves, greater stem diameter and root volume than controls or plants treated with the other chemicals. The data obtained in this study are not conclusive, but indicated certain trends that more glasshouse trials and field trials would resolve. Results presented in this thesis indicate that a major shift has occurred in the status of soilborne fungi associated with roots and crowns of grapevines in nurseries in the Western Cape since the 1970s when Phytophthora and Pythium were predominant. The prevalence and role of soilborne fungi need to be determined so that new appropriate disease management strategies can be developed to limit losses in grapevine nurseries and ensure the sustainable production of healthy plants for the grapevine industry. / AFRIKAANSE OPSOMMING: 'N ONDERSOEK NA GRONDGEDRAAGDE SWAMME GEASSOSIEER MET WORTELS EN KRONE VAN WINGERD IN KWEKERYE Grondgedraagde siektes van wingerd is 'n komplekse probleem waaroor min inligting, beide plaaslik en internasionaal, beskikbaar is. Vorige navorsing in Suid-Afrika het aangedui dat swamme van die genera Phytophthora en Pythium die mees algemene en vernietigende grondgedraagde patogene in kwekerye en wingerde in die Wes-Kaap provinsie is. Die plaaslike wingerdbedryf brei huidiglik uit; nuwe kultivars, metodes en landbouchemikalieë word gebruik wat 'n invloed kan hê op grondgedraagde patogene. Gevolglik het dit noodsaaklik geword om die status van grondgedraagde patogene in wingerdkwekerye weer te bepaal, aangesien inligting in hierdie verband noodsaaklik is vir die ontwikkeling van siekte bestuurspraktyke vir die ontwikkelende plaaslike wingerdbedryf. Grondgedraagde swamgenera geassosieer met wortels en krone van terugsterwende wingerd in kwekerye is bepaal in opnames wat by drie verskillende wingerdkwekerye in die Wes-Kaap provinsie uitgevoer is. Cylindrocarpon, Fusarium, Pythium, en Rhizoctonia spp. is konstant vanuit wortels en krone van terugsterwende wingerdplante in kwekery geïsoleer, Cylindrocladiella spp. en Phytophthora cinnamomi is ongereeld vanuit siek wortels, krone en grond geïsoleer, terwyl Pythium spp. algemeen in meeste gronde voorgekom het. Resultate dui daarop dat die status van grondgedraagde swampatogene in wingerdkwekerye in die Wes- Kaap provinsie oor die laaste 30 jaar verander het. Die DNA filogenie en patogenisiteit van die isolate van Cylindrocladiella is bepaal. Vier spesies van Cylindrocladiella kom voor op wingerd in Suid-Afrika, naamlik C. lageniformis, C. parva, C. peruviana, sowel as 'n nuwe spesie, wat in hierdie studie as C. viticola aangedui is en wat deel is van die C. infestans spesie kompleks. Patogenisiteits proewe was onvoldoende om die patogeniese status van die swam me te bepaal. Tien Fusarium spp. is vanuit wortels en krone van terugsterwende wingerdplante in kwekery geïsoleer, naamlik F. acuminatum, F. anthophilum, F. chlamydosporum, F. equiseti, F. nygamai, F. oxysporum, F. proliferatum, F. scirpi, F. semitectum en F. solani. Die dominante spesies was F. oxysporum, gevolg deur F. proliferatum en F. solani. In pathogenisteitsproewe het F. oxysporum en F. solani gelei tot 'n betekenisvolle laer wortelvolume, droë massa van wortels, lengte en droë massa van nuwe groei en aantal blare, maar het die persentasie chlorotiese blare en graad van wortelvrot verhoog. Fusarium proliferatum het ook gelei tot 'n betekenisvolle afname in lengte en massa van nuwe groei, aantal blare en 'n verhoogde graad van wortelvrot in vergelyking met die kontrole behandelings. Dit wil voorkom asof Fusarium solani meer virulent is as F. oxysporum, gevolg deur F. proliferatum. Hierdie is die eerste aanmelding van F. oxysporum, F. proliferatum en F. solani as patogene van wingerd in Suid-Afrika, en die eerste aanmelding van F. proliferatum as 'n patogeen van wingerd in die wêreld. Phytophthora cinnamomi is konstant teen lae frekwensies vanuit terugsterwende wingerd in kwekerye geïsoleer, alhoewel dit in risosfeer gronde teenwoordig was. Dit is moontlik dat die ekstensiewe gebruik van chemikalieë teen donsskimmel in wingerdkwekerye die wingerdplante kan beskerm teen infeksie deur P. cinnamomi. Die effek van chemikalieë wat gebruik word teen donsskimmel op Phytophthora wortelverrotting van wingerd in kwekerye, is 'n glashuis geëvalueer. Die chemikalieë wat gestoets is, het vir die gemete parameters, tot baie min onderskeibare effek gelei relatief tot die kontrole plante, en daar is afgelei dat die inokulasie tegniek verbetering benodig. Plante wat met fosforiensuur behandel is, het egter geneig om langer te wees met meer blare, 'n groter stamdeursnee en wortelvolume as kontrole plante of plante behandel met ander chemikalieë. Data verkry vanuit die hierdie studie was onvoldoende, maar sekere neigings is aangedui wat deur verdere glashuis- en veldproewe verklaar sal word. Resultate wat in hierdie tesis weergegee is, het aangedui dat 'n algehele verskuiwing in die status van grondgedraagde swamme geassosieer met wortels en krone van wingerd in kwekerye vanaf die 1970s, toe Phytophthora en Pythium die dominante genera was, plaasgevind het. Die voorkoms en rol van grondgedraagde swamme moet bepaal word, sodat nuwe voldoende siektebestuurspraktyke ontwikkel kan word om verliese in wingerdkwekerye te beperk en sodoende die volhoubare produksie van gesonde plante vir die wingerdbedryf te verseker.

Dissertation (PhD)--University of Stellenbosch, 2007. / ENGLISH ABSTRACT: Plants have developed sophisticated means of combating plant diseases. The events that prepare the plant for, and follow plant-pathogenic interactions, are extremely complex and have been the topic of intensive investigation in recent years. These interactions involve a plethora of genes and proteins, and intricate regulation thereof; from the host and pathogen alike. Studying the contribution of single genes and their encoded proteins to the molecular dialogue between plant and pathogen has been a focus of plant molecular biologists. To this end, a gene encoding a polygalacturonase-inhibiting protein (PGIP) was recently cloned from Vitis vinifera. These proteins have the ability to inhibit fungal endopolygalacturonases (ePGs), enzymes which have been shown to be required for the full virulence of several fungi on their respective plant hosts. The activity of PGIP in inhibiting fungal macerating enzymes is particularly attractive for the improvement of disease tolerance of crop species. The VvPGIP-encoding gene was subsequently transferred to Nicotiana tabacum for high-level expression of VvPGIP. These transgenic plants were found to be less susceptible to infection by Botrytis cinerea in an initial detached leaf assay. Also, it was shown that ePG inhibition by protein extracts from these lines correlated to the observed decrease in susceptibility to B. cinerea. This study expands on previous findings by corroborating the antifungal nature of the introduced PGIP by whole-plant, timecourse infection assays. Six transgenic tobacco lines and an untransformed wildtype (WT) were infected and the lesions measured daily from day three to seven, and again at day 15. The transgenic lines exhibited smaller lesions sizes from three to seven days post-inoculation, although these differences only became statistically significant following seven days of incubation. At this point, four of the six lines exhibited significantly smaller lesions than the WT, with reductions in disease susceptibility ranging between 46 and 69% as compared to the WT. Two of the lines exhibited disease susceptibility comparable to the WT. In these resistant plant lines, a correlation could be drawn between Vvpgip1 expression, PGIP activity and ePG inhibition. These lines were therefore considered to be PGIP-specific resistant lines, and provided ideal resources to further study the possible in planta roles of PGIP in plant defense. The current hypothesis regarding the role(s) of PGIP in plant defense is twofold. Firstly, PGIPs have the ability to specifically and effectively inhibit fungal ePGs. This direct inhibition results in reduced fungal pathogenicity. Alternatively, unhindered action of these enzymes results in maceration of plant tissue and ultimately, tissue necrosis. Subsequently, it could be shown that, in vitro, the inhibition of ePGs prolongs the existence of oligogalacturonides, molecules with the ability to activate plant defense responses. Thus, PGIPs limit tissue damage by inhibition of ePG; this inhibition results in activation of plant defense responses aimed at limiting pathogen ingress. Several publications reported reduced susceptibility to Botrytis in transgenic plant lines overexpressing PGIP-encoding genes. However, none of these publications could expand on the current hypotheses regarding the possible in planta roles of PGIP in plant defense. In this study we used transgenic tobacco lines overexpressing Vvpgip1 as resources to study the in planta roles for PGIP. Transcriptomic and hormonal analyses were performed on these lines and a WT line, both before and following inoculation with Botrytis cinerea. Transcriptomic analysis was performed on uninfected as well as infected tobacco leaf material utilizing a Solanum tuberosum microarray. From the analysis with healthy, uninfected plant material, it became clear that genes involved in cell wall metabolism were differentially expressed between the transgenic lines and the WT. Under these conditions, it could be shown and confirmed that the gene encoding tobacco xyloglucan endotransglycosylase (XET/XTH) was downregulated in the transgenic lines. Additionally, genes involved in the lignin biosynthetic pathway were affected in the individual transgenic lines. Biochemical evidence corroborated the indication of increased lignin deposition in their cell walls. Additionally, phytohormone profiling revealed an increased indole-acetic acid content in the transgenic lines. These results show that constitutive levels of PGIP may affect cell wall metabolism in the Vvpgip1-transgenic lines which may have a positive impact on the observed reduced susceptibilities of these plants. An additional role for PGIP in the contribution to plant defenses is therefore proposed. PGIP may directly influence defense responses in the plant leading to the strengthening of cell walls. This might occur by virtue of its structural features or its integration in the cell wall. These reinforced cell walls are thus “primed” before pathogen ingress and contribute to the decrease in disease susceptibility observed in lines accumulating high levels of PGIP. Transcriptional and hormonal analyses, at the localized response, were performed on Botrytis-infected leaf tissue of the transgenic lines and a WT line. Several Botrytis responsive genes were found to be upregulated in both the WT and the transgenic lines. Although limited differential expression was observed between the two genotypes, the analyses identified a gene which was upregulated two-fold in the transgenic lines, as compared to WT. This was confirmed by quantitative Real-Time PCR. This gene is involved in the lipoxygenase pathway, specifically the 9-LOX branch, leading to the synthesis of the divinyl ether oxylipins colneleic and colnelenic acid, which show inhibitory effects on Botrytis spore germination. Phytohormone profiling revealed that the transgenic lines accumulated more of the defense-related hormone pool of jasmonates. These are formed via the 13-LOX pathway and have been shown to be important for the restriction of Botrytis growth at the site of infection. Collectively, the results from the infection analyses indicate that in these transgenic lines, both branches of the lipoxygenase pathway are differentially induced at the level of the localized response to Botrytis infection. Similarly, an increased induction of the synthesis of the defense-related hormone salicylic acid could be observed, although this hormone did not accumulate to significantly higher levels. These results are the first report of differential induction of a defense-related pathway in pgip-overexpressing lines and substantiate the proposal that following ePG inhibition by PGIP, signaling which activates plant defense responses, takes place. Taken together, these results significantly contribute to our understanding of the in planta role of PGIP in plant defense responses. / AFRIKAANSE OPSOMMING: Plante het deur evolusie gesofistikeerde meganismes teen die aanslag van plantsiektes ontwikkel. Die gebeure wat die plant voorberei, asook dié wat op plant-patogeen interaksies volg, is uiters kompleks en vorm die kern van verskeie navorsingstemas die afgelope paar jaar. Etlike plant- én patogeengene en proteïene is by hierdie interaksies betrokke en aan komplekse reguleringsprosesse onderworpe. Die bestudering van die bydrae van enkelgene en hul gekodeerde proteïene tot die molekulêre interaksie tussen ‘n plant en patogeen is ‘n sterk fokus van plant-molekulêre bioloë. Met hierdie doel as fokus, is ‘n geen wat vir ‘n poligalakturonaseinhiberende proteïen (PGIP) kodeer, van Vitis vinifera gekloneer. Hierdie proteïene beskik oor die vermoë om fungiese endopoligalakturonases (ePG's), ensieme wat benodig word vir die virulensie van verskeie fungi op hul gasheerplante, te inhibeer. Die inhibisie van ePG's deur PGIP en die gepaardgaande verminderde weefseldegradasie is ‘n baie belowende strategie vir die verbetering van verboude gewasse se patogeentoleransie. Die VvPGIPenkoderende geen is gevolglik na Nicotiana tabacum oorgedra vir hoëvlakuitdrukking van VvPGIP. Daar is gevind dat hierdie transgeniese plante minder vatbaar vir Botrytis cinerea-infeksies was in ‘n inisiële antifungiese toets wat gebruik gemaak het van blaarweefsel wat van die moederplant verwyder is. Daar is ook ‘n korrelasie gevind tussen B. cinerea-siekteweerstand en ePG-inhibisie deur proteïenekstrakte van die transgeniese populasie. Die huidige studie bou voort op en bevestig vorige bevindinge betreffende die antfungiese aard van die heteroloë PGIP in die heelplant en oor tyd. Ses transgeniese tabaklyne en 'n ongetransformeerde wilde-tipe (WT) is geïnfekteer en die lesies is vanaf dag drie tot sewe, en weer op dag 15, gemeet. Die transgeniese lyne het in die tydperk van drie tot sewe dae ná-inokulasie kleiner lesies as die WT getoon, alhoewel hierdie verskille slegs statisties beduidend geword het na sewe dae van inkubasie. Op daardie tydstip het vier van die ses lyne aansienlik kleiner lesies as die WT getoon, en verlagings in siektevatbaarheid het, in vergelyking met die WT, van 46% tot 69% gewissel. Twee van die lyne het siektevatbaarheid getoon wat vergelykbaar was met dié van die WT. In die siekteweerstandbiedende plantlyne was daar 'n verband tussen Vvpgip1-ekspressie, PGIP-aktiwiteit en ePG-inhibisie. Hierdie plantlyne is dus as PGIP-spesifieke siekteweerstandslyne beskou en dien dus as ideale eksperimentele bronne vir die ontleding van die moontlike in plantafunksies van PGIP in plantsiekteweerstandbiedendheid. Die huidige hipotese betreffende die funksie(s) van PGIP in plantsiekteweerstand is tweeledig. Eerstens het PGIP die vermoë om fungusePG's spesifiek en doeltreffend te inhibeer. Hierdie direkte inhibisie veroorsaak ‘n vermindering in patogenisiteit van die fungus op die gasheer. Indien ePG's egter hulle ensimatiese aksie onverstoord voortsit, sal weefseldegradasie en uiteindelik weefselnekrose die gevolg wees. Daar kon ook bewys word dat die in vitroinhibisie van ePG's deur PGIP die leeftyd van oligogalakturoniede, molekules wat die vermoë het om die plantweerstandsrespons aan te skakel, kan verleng. PGIP het dus nie net die vermoë om ePG's, en dus weefseldegradasie, te inhibeer nie; maar hierdie inhibisie lei ook daartoe dat plantweerstandsresponse aangeskakel word met die oog op die vermindering van patogeenindringing. Verskeie publikasies het reeds gerapporteer oor verminderde Botrytisvatbaarheid in PGIP transgeniese plantlyne. Geeneen van hierdie publikasies kon egter uitbrei op die huidige hipotese aangaande die moontlike in planta-funksie van PGIP in plantsiekteweerstand nie. In hierdie studie is transgeniese tabaklyne wat PGIP ooruitgedruk gebruik om hierdie moontlike in planta-funksies vir PGIP uit te klaar. Transkriptoom- en hormonale analises is op hierdie plantlyne en ‘n WT voor en ná inokulasie met die nekrotroof Botrytis cinerea uitgevoer,. Transkriptoomanalises is uitgevoer op ongeïnfekteerde, sowel as geïnfekteerde tabakblaarmateriaal deur gebruik te maak van ‘n Solanum tuberosum-mikroraster. Die analises met gesonde, ongeïnfekteerde plantmateriaal het daarop gewys dat gene betrokke by selwandmetabolisme tussen die transgeniese lyne en die WT verskillend uitgedruk was. Dit kon bewys word dat, sonder infeksiedruk, die geen wat xiloglukaan-endotransglikosilase (XET) kodeer, in die transgeniese lyne afgereguleer was. Gene wat betrokke is in die lignien-biosintetiese pad was ook in die individuele transgeniese lyne beïnvloed. Biochemiese toetse het ook die aanduiding van verhoogde ligniendeposisie in die transgeniese lyne se selwande bevestig. Addisionele fitohormoonprofiele het getoon dat hierdie lyne ook beskik oor verhoogde vlakke van indoolasynsuur (IAA). Hierdie resultate wys daarop dat konstitutiewe vlakke van PGIP selwandmetabolisme in die Vvpgip1-transgeniese lyne moontlik kan beïnvloed, wat plantsiekteweerstand in dié lyne positief kan beïnvloed. Dit wil dus voorkom asof PGIP 'n bykomende funksie in plantsiekteweerstand het. Plantweerstandsreponse kan direk deur PGIP beïnvloed word, wat tot die versterking van plantselwande kan lei; dit kan geskied by wyse van die strukturele eienskappe van die proteïen of die integrasie daarvan in die selwand. Hierdie selwande is dus “voorberei” alvorens patogeenindringing plaasvind en kon bydra tot die verminderde siektevatbaarheid wat waargeneem is in lyne wat hoë vlakke van PGIP akkumuleer. Transkriptoom- en hormonale analises is ook uitgevoer op Botrytisgeïnfekteerde blaarmateriaal van beide die transgeniese lyne en ‘n WT. Verskeie Botrytis-responsgene is in beide die transgeniese lyne en die WT opgereguleer. Differensïele geenekspressie tussen die twee genotipes was taamlik beperk, maar in die analises kon ‘n geen geïdentifiseer word wat tweevoudig in die transgeniese lyne opgereguleer was in vergelyking met die WT. Hierdie resultaat is ook bevestig met behulp van die “Real-Time” Polimerasekettingreaksie (PKR). Hierdie geen is betrokke in die lipoksigenase (LOX) -pad (spesifiek die 9-LOXarm), wat tot die sintese van die diviniel-eter oksilipiene “colneleic-” en “colnelenic”-suur lei. Daar is al bewys dat hierdie twee verbindings Botrytisspoorontkieming kan inhibeer. Fitohormoonprofiele van die geïnfekteerde plante het gewys dat die transgeniese lyne verhoogde vlakke van die poel van jasmonate wat plantsiekteweerstands-hormone is, ná inokulasie akkumuleer. Hierdie hormone word in die 13-LOX-arm van die lipoksigenase pad gevorm en is belangrik vir die beperking van Botrytis by die infeksiesetel. Die resultate van die analises wat op Botrytis-infeksie volg, dui daarop dat beide arms van die lipoksigenasepad in die transgeniese lyne verskillend by die lokale respons geïnduseer word. ‘n Verhoogde induksie van ‘n ander plantsiekteweerstandshormoon, salisielsuur, kon ook opgemerk word, alhoewel die totaal geakkumuleerde vlakke nie beduidend hoër was as dié van die WT nie. Hierdie resultate is die eerste wat onderskeidende induksie van ‘n siekteweerstandspad in enige van die pgip-ooruitgedrukte plantlyne rapporteer. Daarmee ondersteun dit ook die hipotese dat, seintransduksie wat plantweerstandsresponse aanskakel, ná inhibisie van ePG deur PGIP plaasvind. Die resultate wat met hierdie studie verkry is, dra dus beduidend by tot die huidige kennis van die in planta-funksie van PGIP in plantsiekteweerstandsresponse.

Thesis (MScAgric (Soil Science))--University of Stellenbosch, 2005. / This thesis forms part of a research program that is investigated by ARC Infruitec- Nietvoorbij and the University of Stellenbosch. The aim of this program is to quantify the effect of soil and climate on wine quality and character as a basis for scientific demarcation of production regions. Natural factors are first quantified and characterised so that relative homogeneous natural terroir units (NTU) can be identified. Thereafter wines made from these different localities are assessed for similarities which permit grouping of localities into viticultural terroirs. The present study involved seven research plots in the Robertson district, making use of initial measurements by ARC Infruitec-Nietvoorbij in the 2000/01 and 2001/02 seasons together with more intensive measurements in the 2002/03 and 2003/04. The seven plots are located in two wards, Le Chasseur and Vinkrivier. Sauvignon blanc plots were marked at two localities, each with two sub-plots occurring in one block. Three Cabernet Sauvignon plots were marked out. Two plots occurred in one block in Le Chasseur and one plot in Vinkrivier. Plots within the same block were located on different soil types so that the effect of soil type on wine quality and character could be monitored. Morphological examination of soil profiles at each plot was carried out together with physical and chemical analysis of samples taken at 30cm depth intervals. Soil water content was measured at regular intervals to assess soil water tension. Vineyard performance was assessed in terms of phenology, root system studies, leafwaterpotential measurements, canopy quality assessment, leaf analysis, shoot growth, and data relating to ripening, harvest and pruning. Wines were made from each plot by ARC Infruitec- Nietvoorbij and evaluated by a tasting panel after six months and then again after eighteen months. Meteorological data were collected in each ward.

Thesis (MscAgric (Viticulture and Oenology))--University of Stellenbosch, 2009. / ENGLISH ABSTRACT: Little scientific information regarding the effect of different geological parent materials on grapevine performance is currently available in South Africa. This aspect is of special significance for the Helderberg area, where parent material may change from granite to shale over a short distance. This results in shale- as well as granite-derived soils often occurring within the same vineyard. The objectives of this study were to (i) quantify the nutritional status and other soil properties of different parent materials (shale and granite) and overlying soils (ii) investigate the impact of geological differences in the soil on the vine nutritional status and certain vine parameters. This study was done over two seasons (2006/2007 and 2007/2008). Two Sauvignon blanc and two Cabernet Sauvignon vineyard blocks were selected at two different localities for each cultivar in the Helderberg area, South Africa. Shale- and granitederived soils were identified within each block. Kaolinite was the dominant mineral, whereas quartz and feldspar were sub-dominant. Traces of mica were also present in some shale- and granite-derived soils. Granite- contained significantly higher contents of coarse sand than shale-derived soils, whilst the opposite occurred in terms of fine sand. These differences affected the water holding capacity, in general making it higher in shale- than granite-derived soils. Shale-derived soils had higher concentrations of total K but granite-derived soils had a higher ability to release K as they contained higher concentrations of soluble K. The Q/I parameters, potential buffering capacity of K (PBCK) and equilibrium activity ratio of K (ARK) showed no consistent responses to geological differences. Potassium concentrations were higher in the leaf blades (obtained before harvest in 2007) from Sauvignon blanc grapevines on granite- than on shale-derived soils. Potassium concentrations in the Cabernet Sauvignon juice (obtained in 2007) tended higher in juice from granite- than from shale-derived soils. In 2008, K concentrations tended higher in juice from shale- than from granite-derived soils for both cultivars. The pH of the Sauvignon blanc juice (obtained in 2008) tended higher in juice from shale-than from granite-derived soils, thus corresponding with the K concentrations in the juice in this season. Nitrogen concentrations were higher in Cabernet Sauvignon juice (obtained in 2007) and Sauvignon blanc juice (obtained in 2008) from shalethan from granite-derived soils. In terms of vine water status, vines on granite-derived soils appeared more stressed than those on shale-derived soils in both seasons at one of the vineyards. In these Sauvignon blanc and Cabernet Sauvignon vineyards, the K nutritional status was not affected by geology in a consistent manner but there were some noticeable tendencies for a specific cultivar during certain seasons. On account of vines being planted on shale- and granite-derived soils within the same block, soil preparation was done similarly for both soils, and they were exposed to similar irrigation schedules, canopy management strategies and climatic conditions. Therefore, there is a high probability that all these practices may have negated the effect of geology on the K status of soils and especially on juice K concentration at the time of harvest. It was clear that seasonal differences and fertilisation affected the nutritional status of the vines more than geology. / AFRIKAANSE OPSOMMING: In Suid-Afrika is daar tans min wetenskaplike inligting oor die effek van verskillende geologiese moedermateriale op die prestasie van wingerd beskikbaar. Hierdie aspek is veral van belang in die Helderberg-area, waar moedermateriaal oor ‘n baie kort afstand van graniet na skalie kan wissel. Dit lei daartoe dat skalie-, sowel as granietgronde, dikwels binne dieselfde wingerd voorkom. Die doelwitte van die studie was om: (i) die voedingstatus en ander grondkundige eienskappe van die verskillende moedermateriale (skalie en graniet) en oorliggende gronde te kwantifiseer (ii) die impak van geologiese verskille in die grond op wingerd se voedingstatus en sekere wingerdkundige parameters, te ondersoek. Hierdie studie is oor twee seisoene (2006/2007 en 2007/2008) gedoen. Twee Sauvignon blanc en twee Cabernet Sauvignon wingerdblokke is geselekteer by twee verskillende lokaliteite vir elke kultivar in die Helderbergarea, Suid-Afrika. Beide skalie- en granietgrond is binne elke blok geïdentifiseer. Kaoliniet was die dominante mineraal, met kwarts en veldspaat sub-dominant, terwyl spore van mika ook in beide skalie- en granietgronde gevind is. Granietgronde het betekenisvol hoër hoeveelhede growwe sand bevat, terwyl skaliegronde meer fyn sand bevat het. Hierdie verskille het ‘n effek op waterhouvermoë gehad en daartoe gelei dat waterinhoude oor die algemeen hoër was vir skaliegronde. Skaliegronde het groter hoeveelhede totale K bevat, maar granietgronde se vermoë om K vry te stel was hoër, omdat hulle ‘n hoër konsentrasie oplosbare K bevat het. Die Q/I parameters, potensiële buffervermoë vir K (PBCK) en ewewig aktiwiteitsverhouding vir K (ARK), is nie op ‘n konsekwente wyse deur geologiese verskille beïnvloed nie. Vir die Sauvignon blanc wingerde was kalium konsentrasies in blaarskywe (gemonster voor oes in 2007) hoër vir graniet- as vir skaliegronde. Kalium konsentrasies in die sap vanaf Cabernet Sauvignon (gemonster in 2007) het hoër geneig vir granietgronde. In 2008 het die kalium konsentrasies, vir beide kultivars, hoër geneig in sap vanaf skaliegronde. Gedurende dié seisoen het die pH van sap ook hoër geneig vir Sauvignon blanc vanaf skaliegronde, wat dus ooreenstem met die K inhoud van die sap. Stikstof konsentrasies was hoër in sap vanaf skaliegronde vir Cabernet Sauvignon (2007) en vir Sauvignon blanc (2008). In terme van die wingerde se waterstatus, het stokke op die granietgrond, by een van die lokaliteite, geneig om gedurende beide seisoene onder groter stremming te wees op graniet as op skaliegrond. In hierdie Sauvignon blanc en Cabernet Sauvignon wingerde, is K voedingstatus nie op ‘n konsekwente wyse deur geologie geaffekteer nie, maar gedurende sommige seisoene was daar wel duidelike tendense vir ‘n spesifieke kultivar. Omdat die stokke binne dieselfde blok op skalie- en graniet gronde geplant is, was grondvoorbereiding eenders vir die twee grondtipes terwyl besproeiingskedule, lowerbestuur en klimaatstoestande ook identies was. Daar is dus ‘n hoë waarskynlikheid dat al hierdie faktore daartoe kon bygedra het dat die effek van geologie op die K status van van gronde versluier is, veral die effek op die K inhoud van sap teen oestyd. Dit was duidelik dat seisoenale klimaatsverskile en bemestingspraktyke ’n groter effek as geologie op die voedingstatus van die wingerd gehad het.

Thesis (PhD (Agric) (Conservation Ecology and Entomology))--University of Stellenbosch, 2006. / A study was performed to develop a generic pest monitoring system for sampling the main table grape pests in vineyards in the Hex River Valley, Western Cape Province of South Africa. The presence of phytophagous and predatory mites on cover crop plants was also investigated as this may contribute to biological control of the phytophagous mites in vines. Life table studies for Epichoristodes acerbella (Walker), an important phytosanitary pest, were conducted to determine whether or not this pest was sensitive to high temperatures. Information gained from the latter can also be used for breeding purposes in the possible future development of a sterile insect technique (SIT) programme to control this pest. The sampling system consisted of inspecting 20 plots of five vines per plot per one to two hectares. The top fork of each of the five vines per plot was examined for Planococcus ficus (Signoret) to a distance of within 30 cm of the stem, as well as the distal 15 cm of one cane per vine for the presence of P. ficus and damage caused by Phlyctinus callosus Boh. One bunch per vine was examined for insect damage or presence, and one leaf per vine for the presence of leaf infesting arthropods, such as Tetranychus urticae Koch, P. ficus and Frankliniella occidentalis (Pergande). Corrugated cardboard bands, tied around the stem of one vine per plot, were used to monitor activity of P. callosus. Blue sticky traps, at a density of four to five traps per one to two hectares, were used to monitor activity of F. occidentalis. Pheromone traps, at a density of one trap per one to two hectares, were used to monitor activity of P. ficus, E. acerbella and Helicoverpa armigera (Hübner). All the above-mentioned inspections were done at two-weekly intervals, except traps for E. acerbella and H. armigera, which were inspected weekly. In each of the rows in which the sample plots were situated, one leaf of each of the cover crop plant species was examined for the presence of phytophagous mites and their predators. The abundance and distribution of cover crop plants were determined using a co-ordinate sampling system. Cover crop sampling was done at monthly intervals. The current threshold for P. ficus is 2% stem infestation, which is reached when more than 65 males per pheromone trap are recorded. Counting mealybugs on the sticky pads in the pheromone traps is time consuming. However, the number of grid blocks on the sticky pad with males present can be counted. When P. ficus males are found in 27 blocks on the sticky pad, stem inspections should commence. Due to the spatial association between P. ficus bunch and stem infestation, stem infestation could give an indication of where bunch infestation could be expected. The use of blue sticky traps for predicting halo spot damage, caused by F. occidentalis, is not recommended. The presence of thrips on the vine leaves could not give an indication of where to expect bunch damage, since thrips on the leaves and halo spot damage were not spatially associated. A suitable sampling method for F. occidentalis still needs to be developed. The monitoring system described here can only provide information on the infestation status of the vineyard. For E. acerbella, H. armigera and P. callosus, the traps and cardboard bands could be used to identify vineyards where these pests are present and therefore, where phytosanitary problems may arise. The presence of P. callosus under the bands was spatially associated with P. callosus damage and could be used as an indicator of the latter. The presence of drosophilid flies in the bunches could not be used as an indicator of the presence of E. acerbella in the bunches. If 5% bunch damage is used as an economic threshold for E. acerbella and P. callosus, there will be a good chance of not under spraying if control measures are applied at 1% bunch damage. Epichoristodes acerbella favoured more moderate constant temperatures, with constant temperatures of 28°C and above being unfavourable for development. The economic threshold for Tetranychus urticae Koch is six mites per leaf, or if presence-absence sampling is used, 11 to 29% leaf infestation. Three important predatory mites, that kept T. urticae under control, were found in the Hex River Valley, namely Euseius addoensis (Van der Merwe & Ryke), Neoseiulus californicus (McGregor) and an undescribed phytoseiid in the genus Typhlodromus. Various cover crop plants served as hosts for T. urticae and predatory mites. The presence of these plants created suitable conditions for the survival of these mites and may have influenced their presence on the vine leaves. In the case of phytosanitary pests, both field and pack shed inspections can be used to conclude with a 99% degree of certainty that infestation levels in the pack shed will be 10% or less, since similar results for both methods were obtained. However, more than 20 plots will have to be inspected.

Thesis (MSc (Genetics))--Stellenbosch University, 2008. / South Africa is one of the top ten wine producing countries in the world. The South African wine industry contributes approximately R16.3 billion to South Africa’s annual gross domestic product with 42.8% of wine being exported. To compete with the top wine producing countries and to ensure a viable export market, South Africa needs to ensure that healthy, virus free propagation material is produced and sold. One of the viruses that need to be tested for is Grapevine fanleaf virus (GFLV). Grapevine fanleaf virus causes degeneration and malformation of berries, leaves and canes and is responsible for significant economic losses by reducing crop yields by as much as 80%, reducing the longevity of the vines and affecting fruit quality. It is widespread in the Breede River Valley of the Western Cape where the nematode vector, Xiphinema index, is prevalent. The Breede River Valley contributes approximately 30% of the total production of the local wine industry, and severe losses in this region could threaten the viticulture. The Plant Improvement Act states that all propagation material sold must be tested for GFLV by a reputable scientific technique. The technique commonly used in South Africa is the Double Antibody Sandwich - Enzyme-linked Immunosorbent Assay (DAS-ELISA) and the kits are imported from Europe at a significant cost to the South African viticulture industry. The objective of this study was to produce a reliable and sensitive diagnostic assay specific for the South African strains of GFLV. This project aimed to develop and optimize a DAS-ELISA, by using recombinant DNA technology to produce antibodies against bacterially expressed viral coat protein. Total RNA was extracted from GFLV infected grapevine material and the viral coat protein (CP) amplified. The CP was cloned into the pGex-6P-2 expression vector, fusing a Glutathione STransferase (GST) partner to the viral coat protein enhancing solubility and protein purification. Insufficient amounts of the soluble protein were expressed and purified, preventing the production of antibodies and thus the development of the DAS-ELISA. An alternative diagnostic rapid-direct-one-tube-RT-PCR assay was developed. This rapid-directone- tube-RT-PCR assay was compared to commercially available DAS-ELISA and ImmunoStrip tests (Agdia) to assess the reliability, sensitivity and specificity of the rapid-direct-one-tube-RTPCR assay. Twelve GFLV isolates from South Africa were sequenced to investigate the variability between the isolates as well as the variability between the South African isolates and GFLV sequences available in Genbank. Sequence identities between clones from different GFLV isolates from South Africa were between 86-99% and 94-99% at the nucleotide and amino acid levels, respectively. Phylogenetic analysis based on the coat protein gene sequences showed that the South African isolates form two distinct clades or sub-populations. No significant correlation was found between geographical origin and symptoms, nor between geographical origin and sequence variability or between grapevine cultivar and symptom expression. Of the 23 samples tested with all three tests, 21 tested positive with rapid-direct-one-tube-RT-PCR, 19 with the ImmunoStrips and 17 with an imported DAS-ELISA kit (Agdia). Rapid-direct-one-tube-RT-PCR was found to be the most reliable technique for GFLV detection. Although the establishment of a DAS-ELISA directed to the South African strain(s) of GFLV was not successful, an alternative PCR based diagnostic system was developed, and proved to be sensitive and reliable. RT-PCR based diagnostic assays are generally accepted to be more sensitive than DAS-ELISA, but the latter is still used as the diagnostic assay of choice for routine testing due to ease of use. This rapid-direct-one-tube-RT-PCR assay is a rapid, sensitive and reliable diagnostic test, reducing the prevalence of false negatives, contributing to a virus free viticulture industry. The rapid-direct-one-tube-RT-PCR assay is as easy to use as DAS-ELISA, faster and can be performed by semi skilled workers, thus providing all the advantages associated with DAS-ELISA.