Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: Within-vineyard variability in vigour and water status commonly occurs in South African vineyards. Different soil types found over short distances are probably the main cause of vigour variability, while differences in grapevine water status are commonly induced by lateral water flow in the vineyard, blocked irrigation emitters and differences in soil water-holding capacity. These factors can cause heterogeneous ripening and differences in fruit quality between different parts of the vineyard, an aspect that needs to be avoided as far as possible in order to produce quality wines. Measurements of carbon isotope discrimination (CID) have proved to be a tool to assess grapevine physiology in order to study the effects of environmental parameters on leaf carbon dioxide (CO2) gas exchange and stomatal conductance (gs). Grapevine water deficit stress/strain in reaction to these environmental conditions can then be determined by observing the amount of 13C absorbed by plant material after discrimination of 13C has taken place, and this is influenced by the grapevine stress condition and can indicate water-use efficiency. In this study, the variability of grapevine water status and vigour was determined in order to quantify these parameters in different parts of the vineyard. Two separate trials were conducted, the first at Wellington, South Africa, where different irrigation regimes resulted in variability in grapevine water status between plots. The second trial was at Stellenbosch, South Africa, where plots were divided among different vigour classes and irrigation was applied in different quantities for different irrigation treatments. Within-vineyard variability in water status (Wellington and Stellenbosch) and vigour (Stellenbosch) were then quantified and the effects on some grapevine physiological parameters and berry composition were measured. The treatments in the Wellington trial led to differences in grapevine water status, which could be quantified by measurements of stem water potential (SWP) and leaf water potential (LWP). Soil variability also led to differences in grapevine vigour, which were quantified by measurements of pruning mass, leaf area and shoot length. The effect of the variability in grapevine water status on grapevine physiology was assessed by measuring CID, which was the main focus of the study. Other physiological measurements, such as gs and leaf and canopy temperature, were also conducted. The effect of these conditions on grape berry composition was also studied. In the Stellenbosch trial, soil water content, plant water status measurements (SWP, predawn LWP and LWP), physiological measurements (CID and gs) and berry size measurements were used to classify plots into water status treatments (“wet” and “dry” treatments). The effect of vigour differences was analysed separately from these treatments by using pruning mass as a covariate in the statistical analyses. The effect of vigour variability on the measurements was studied by looking at the effect of the covariate on the measurements, while shoot growth rate, shoot length and leaf area measurements were conducted as vegetative growth measurements. Differences in measurements were then studied between the treatments and between the vigour levels of the different plots. In the Wellington trial, plant water status was determined by irrigation, showing increased stress for treatments that received less irrigation. The differences in plant water status then caused differences in grapevine physiology between the treatments, leading to increased gs for increased irrigation. This of course influenced leaf internal CO2 and therefore CID, although CID was also clearly influenced by berry development. Berry size was influenced by irrigation, with larger berries found in wetter treatments, while berry chemical composition was influenced by the irrigation regime, with increased irrigation leading to increased pH and leading to trends showing increased total soluble solids and malic acid, and reduced total and tartaric acid and colour intensity. In the Stellenbosch trial, plots with higher vigour had increased shoot growth rate, longer shoots and increased leaf area, although topping influenced this. Wet treatment vines also showed slightly longer shoots and larger leaf areas. There were differences in soil water content between the wet and dry treatments, and this led to differences in plant water status. Vigour also influenced pre-dawn LWP, especially in the 2007 season, as higher-vigour vines struggled more to rehydrate through the night. Differences in plant water potential led to differences in grapevine physiology, with increased gs for vines from the wet treatment, while higher-vigour vines had slightly increased gs. The differences in gs led to gas exchange differences and therefore differences in CID, meaning that water status and vigour influenced CID. CID measurements illustrated the long term effect of water status on plant physiology, while measurements such as SWP illustrated the short term effects. CID measurements therefore proved to be accumulative over the season, in contrast to SWP measurements that were much more dependent on the current state of grapevine water status. Other physiological measurements showed that wet-treatment vines had higher photosynthetic rates and evapotranspiration and lower leaf temperatures, while higher-vigour vines had slightly increased evapotranspiration and decreased leaf temperatures. Wet-treatment vines had larger berries, while a higher vigour also led to slightly larger berries. Berry composition was influenced by treatment, where wet-treatment vines had increased pH and total soluble solids, while higher-vigour vines had increased juice pH and, in the 2008 season, decreased total soluble solids. Extremely stressed conditions did not show significant effects on plant water potential, but SWP measurements indicated slightly higher stress for the extremely stressed vines and LWP showed slightly less stressed conditions for these vines. Measurements of gs showed slightly lower values for the extremely stressed vines, while measurements of CID showed large significant differences, with the extremely stressed vines having measurements showing high stress. The measurement therefore indicated highly stressed conditions accurately, while other physiological measurements, such as photosynthetic rate, evapotranspiration and leaf temperatures, only showed trends and no significant differences. Measurements of stomatal conductance reacted to plant water status measurements throughout the diurnal measurement days, while CID only reacted slightly with gs changes during these days and was perhaps influenced more by berry chemical composition and development at this early stage of the season. Vigour and water status therefore influenced grapevine physiology, with a more direct effect by water status and an indirect effect by vigour due to microclimatic differences. This also influenced berry composition and therefore quality. In future studies, CID measurements should be done on juice from which organic acids have been removed in order to eliminate the effect of seasonal berry composition on the measurement. Measurements of CID proved to be an integrative, but sensitive, indicator of grapevine stress, especially at the end of the season. It might at best be useful as a post-harvest management tool for producers or grape buyers, especially for irrigation control, as has also been stated by Van Leeuwen et al. (2007). / AFRIKAANSE OPSOMMING: Binne-wingerd variasie in groeikrag en waterstatus is algemeen in Suid-Afrikaanse wingerde. Verskillende grondsoorte wat na aan mekaar voorkom, is seker een van die vernaamste oorsake van variasie in groeikrag, terwyl verskille in wingerdwaterstatus algemeen deur laterale watervloei in die wingerd, verstopte besproeiingspuite en verskille in grond waterhouvermoë geïnduseer word. Hierdie faktore kan aanleiding gee tot heterogene rypwording en verskille in vrugkwaliteit tussen verskillende dele van die wingerd, ‘n aspek wat so ver moontlik vermy moet word om kwaliteitwyne te kan produseer. Die meting van koolstof-isotoopdiskriminasie (KID) is bewys om as gereedskap te kan dien vir die assessering van wingerdfisiologie om die effekte van omgewingsparameters op blaar koolstofdioksied (CO2) - gasuitruiling en stomatale geleiding (gs) te bestudeer. Die stres/stremming as gevolg van ‘n watertekort in die wingerd in reaksie op hierdie omgewingstoestande kan dan bepaal word deur te kyk na hoeveel 13C deur die plantmateriaal geabsorbeer word ná 13C-diskriminasie plaasgevind het, en dít word deur die wingerdstrestoestande beïnvloed en kan ‘n aanduiding verskaf van die doeltreffendheid van waterverbruik. In hierdie studie is die variasie in wingerdwaterstatus en groeikrag bepaal om hierdie parameters in verskillende dele van die wingerd te kwantifiseer. Twee afsonderlike proewe is uitgevoer, die eerste by Wellington, Suid-Afrika, waar verskillende besproeiingsregimes gelei het tot verskille in die wingerdwaterstatus tussen persele. Die tweede proef was by Stellenbosch, Suid-Afrika, waar persele tussen verskillende groeikragklasse verdeel is en besproeiing in verskillende hoeveelhede vir verskillende besproeiingsbehandelings toegepas is. Binne-wingerd variasie in waterstatus (Wellington en Stellenbosch) en groeikrag (Stellenbosch) is toe gekwantifiseer en die effekte op sekere wingerd-fisiologiese parameters en korrelsamestelling is gemeet. Die behandelings in die Wellington-proef het gelei tot verskille in wingerdwaterstatus, wat deur metings van stamwaterpotensiaal (SWP) en blaarwaterpotensiaal (BWP) gekwantifiseer kon word. Grondverskille het ook gelei tot verskille in wingerdgroeikrag, wat deur metings van snoeimassa, blaaroppervlak en lootlengte gekwantifiseer is. Die effek van die variasie in wingerdwaterstatus op wingerdfisiologie is deur metings van KID bepaal wat die hooffokus van hierdie studie was. Ander fisiologiese metings, soos gs en blaar- en lowertemperatuur, is ook gedoen. Die effekte van hierdie toestande op die samestelling van die druiwekorrels is ook bestudeer. In die Stellenbosch-proef is grondwaterinhoud, metings van plantwaterstatus (SWP, voorsonopgang SWP en BWP), fisiologiese metings (KID en gs) en metings van korrelgrootte gebruik om die persele in waterstatusbehandelings (“nat” en “droë” behandelings) te verdeel. Die effek van verskille in groeikrag is apart van hierdie behandelings geanaliseer deur snoeimassa as ‘n kovariaat in die statistiese analises te gebruik. Die effek van groeikragvariasie op die metings is bestudeer deur ondersoek in te stel na die effek van die kovariaat op die metings, terwyl lootgroeitempo-, lootlengte- en blaaroppervlakmetings as metings van vegetatiewe groei uitgevoer is. Verskille in metings tussen die behandelings en tussen die groeikragvlakke van die verskillende persele is toe bestudeer. In die Wellington-proef is plantwaterstatus deur besproeiing bepaal, met verhoogde stres in behandelings waar daar minder besproeiing toegedien is. Die verskille in plantwaterstatus het dan verskille in wingerdfisiologie tussen die behandelings veroorsaak, wat gelei het tot ‘n verhoogde gs in die geval van verhoogde besproeiing. Dit het natuurlik ‘n effek op die interne CO2 van die blaar en dus op KID gehad, hoewel KID ook duidelik deur korrelontwikkeling beïnvloed is. Korrelgrootte is deur besproeiing beïnvloed, met groter korrels in die natter behandelings, terwyl die chemiese samestelling van die korrel deur besproeiingsregime beïnvloed is. Verhoogde besproeiing het pH verhoog en gelei na tendense wat verhoogde totale oplosbare vaste stowwe en appelsuur, en verminderde totale suur, wynsteensuur en kleurintensiteit getoon het. In die Stellenbosch-proef het persele met hoër groeikrag ook verhoogde lootgroeitempo, langer lote en verhoogde blaaroppervlak getoon, hoewel dit deur top beïnvloed is. Wingerdstokke van die nat behandeling het ook effe langer lote en groter blaaroppervlakke getoon. Daar was verskille in grondwaterinhoud tussen die nat en droë behandelings en dit het verskille in plantwaterstatus veroorsaak. Groeikrag is ook deur voor-sonopgang BWP beïnvloed, veral in die 2007-seisoen, aangesien stokke met hoër groeikrag meer gesukkel het om in die nag te rehidreer. Verskille in plantwaterpotensiaal het gelei tot verskille in wingerdfisiologie, met ‘n verhoogde gs vir stokke in die nat behandeling, terwyl stokke met hoër groeikrag ‘n effens verhoogde gs getoon het. Die verskille in gs het gelei tot verskille in gasuitruiling en dus verskille in KID, wat beteken dat waterstatus en groeikrag ‘n invloed op KID het. KID was meer verteenwoordigend van die langtermyneffekte van water status op plantfisiologie, terwyl metings soos SWP die korttermyneffekte weerspieël het. KID metings was dus akkumalatief oor die seisoen, terwyl SWP metings meer ‘n weerspieëling was van die huidige toestand van plantwaterpotensiaal. Ander fisiologiese metings het getoon dat stokke in die nat behandeling ‘n hoër fotosintesetempo en evapotranspirasie sowel as laer blaartemperature ondervind het, terwyl die stokke met hoër groeikrag effe verhoogde evapotranspirasie en verminderde blaartemperature getoon het. Stokke in die nat behandeling het groter korrels gehad, terwyl hoër groeikrag ook effens groter korrels veroorsaak het. Korrelsamestelling is deur die behandelings beïnvloed, met stokke in die nat behandeling wat verhoogde pH en totale oplosbare vaste stowwe getoon het, terwyl stokke met hoër groeikrag verhoogde pH van die sap en verminderde totale oplosbare vaste stowwe (laasgenoemde in die 2008-seisoen) gehad het. Uitermate toestande van stres het geen beduidende effekte op plantwaterpotensiaal getoon nie, hoewel SWP-metings effens hoër stres vir die uitermate gestresde wingerde getoon het en BWP effens minder gestresde toestande vir hierdie stokke getoon het. Metings van gs het effens laer waardes vir die uitermate gestresde stokke getoon, terwyl metings van KID groot noemenswaardige verskille getoon het, met die metings vir die uitermate gestresde wingerde wat hoër stres aangedui het. Dié meting het dus hoogs gestresde toestande akkuraat aangedui, terwyl ander fisiologiese metings, soos tempo van fotosintese, evapotranspirasie en blaartemperature net tendense en nie beduidende verskille aangedui het nie. Metings van stomatale geleiding het dwarsdeur die dae waarop daaglikse metings gedoen is op plantwaterstatusmetings gereageer, terwyl KID net effens met gs-veranderinge op hierdie dae gereageer het en moontlik meer deur die chemiese samestelling en ontwikkeling van die korrel in hierdie vroeë stadium van die seisoen beïnvloed is. Groeikrag en waterstatus het dus wingerdfisiologie beïnvloed, met ‘n meer direkte effek deur waterstatus en ‘n indirekte effek deur groeikrag as gevolg van mikroklimaatsverskille. Dit het ook korrelsamestelling en dus kwaliteit beïnvloed. In toekomstige studies moet KID-metings gedoen word op sap waarvan die organiese sure verwyder is om die effek van seisoenale korrelsamestelling op die meting uit te sluit. Metings van KID is getoon om ‘n integrerende, maar gevoelige, aanduider van wingerdstres te wees, veral aan die einde van die seisoen. Dit is ten beste miskien bruikbaar as naoesbestuursgereedskap vir produsente of druiwekopers, veral vir besproeiingsbeheer, soos ook reeds deur Van Leeuwen et al. (2007) aangedui is.

Thesis (MScAgric (Viticulture and Oenology. Wine Biotechnology))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: Agriculture worldwide is under great pressure to produce enough food in order to sustain the ever-growing world population. Among the many challenges faced by food producers, crop losses and damage caused by fungal plant pathogens is a major problem. The study of fungal pathogens and the interaction between plants and fungi is therefore essential, and has been carried out for many years. Much has been learned in this time, but the full mechanisms of the various modes of fungal attack and plant defence have still not been elucidated. Many fungi rely on the action of cell-wall degrading enzymes (CWDEs) to breach the plant cell wall and facilitate access to the nutrients within. CWDEs are among the very first enzymes to be secreted at the start of fungal attack, and many of them are considered to be essential pathogenesis factors. Endopolygalacturonases (ePGs) are CWDEs that cleave the homogalacturonan stretches of the plant cell wall and are vital virulence factors for a number of fungi, including Botrytis cinerea. An important defence mechanism of plants involves the inhibition of CWDEs in order to halt or slow down the fungal attack. Plant polygalacturonaseinhibiting proteins (PGIPs) are cell wall associated CWDE-inhibiting proteins that specifically act on fungal ePGs. Many different PGIPs from a number of diverse plant species have been described to date. They are known to have differential inhibition capabilities that often result from only a few key amino acid changes within the leucine-rich repeat (LRR) active domains. Previously, the first grapevine PGIP was isolated and characterised from Vitis vinifera cultivar Pinotage (Vvpgip1). This Vvpgip1 gene was overexpressed in the tobacco species Nicotiana tabacum, and was shown to be very effective in reducing the susceptibility of tobacco towards B. cinerea. The combined results confirmed transgene overexpression, increased PGIP activity and a strong resistance response against Botrytis, leading to the characterisation of these lines as having PGIP-specific resistance phenotypes. In a subsequent transcriptomic analysis of these lines it was found that they display differential expression of cell wall metabolism genes and biochemical characteristics that might indicate possible cell wall strengthening compared to wild-type tobacco under uninfecting conditions. The V. vinifera cultivars are all very susceptible to fungal attack, whereas other grapevine species, specifically the North American Vitis species, are known for their strong resistance and even immunity against many fungal pathogens. Thirty seven PGIPs have previously been isolated from these more resistant species. The amino acid sequences of the active domains of these PGIPs were previously aligned with that of VvPGIP1, and the proteins were found to be highly homologous with each other and with VvPGIP1. The different nonvinifera PGIPs separated into 14 subgroups based on their active domain sequences. For this study, one PGIP from each group was selected for functional analysis in tobacco. The selected PGIP-encoding genes were transformed into tobacco by means of Agrobacterium tumefaciens. Analyses of the putatively transformed plantlets were performed to test for transgene presence, transgene expression, and PGIP activity: final transgenic tobacco populations consisting of three to twelve individually transformed lines of nine different nonvinifera PGIPs were obtained. A subset of the resultant transgenic lines was infected with B. cinerea in two independent whole plant infections over 11-14 days in order to investigate the disease resistance afforded by the various PGIPs towards this fungus. A line from the previously characterised VvPGIP1 population was included as reference; all the infections were contrasted to the WT tobacco. All the infected lines overexpressing the non-vinifera PGIPs displayed very strong disease reduction in comparison to the WT control: after initial primary lesion formation, the spread of fungal infection was contained and halted in these lines, while wild-type tobacco plants were severely affected. Although the VvPGIP1 line displayed the characteristic PGIP-defense response, the non-vinifera PGIP plants displayed smaller lesions, indicating very strong resistance phenotypes. The characterised non-vinifera PGIP overexpressing lines, together with the VvPGIP1 line and the WT control were also used to further evaluate the previous observation that overexpression might lead to changes in expression of cell wall genes. Analysis of the expression of a xyloglucan endotransglycosylase (xth) gene in the transgenic population showed that this gene was down-regulated in healthy uninfected tissue from all the transgenic lines tested. This confirmed previous results and have confirmed in all grapevine PGIP overexpressing lines tested so far that this gene is downregulated. XTH is typically involved in cell wall metabolism and specifically in controlling the strength and elasticity of the plant cell wall. From previous work it is known that downregulation of this gene leads to strengthening of the wall. The results obtained in this study showed that the PGIP-specific resistance phenotype seen for VvPGIP1-overexpressing tobacco could be confirmed in transgenic tobacco overexpressing non-vinifera PGIPs from more resistant grapevine species as well. The fact that these PGIPs lines all performed even better than the VvPGIP1 lines in conferring resistance towards B. cinerea provides an interesting angle for further investigation into the structural differences between the non-vinifera PGIPs and VvPGIP1. The transgenic lines are also excellent material to study the in vivo functions of PGIPs further in the context of plant-pathogen interactions. / AFRIKAANSE OPSOMMING: Die landboubedryf is wêreldwyd onder groot druk om genoeg voedsel te produseer vir die groeiende wêreldbevolking. Een van die grootste probleme wat die bedryf ondervind, is die groot skade wat aan gewasse aangerig word deur patogeniese swamme. Dit is dus noodsaaklik om swamme en die interaksie tussen plante en swamme te bestudeer, en dit word al vir jare gedoen. Hoewel daar al baie geleer is in hierdie tydperk, is die volle meganismes van die verskeie maniere hoe swamme aanval en hoe plante hulleself verdedig, nog nie bekend nie. Verskeie swamme maak staat op die aktiwiteit van selwand-afbrekende ensieme (SWAEe) om deur die plantselwand te breek en sodoende toegang tot voedingstowwe in die plantsel te fasiliteer. SWAEe is van die eerste ensieme wat tydens die begin van patogeniese aanval deur swamme afgeskei word en verskeie SWAEe word as noodsaaklike patogeniese faktore beskou. Endopoligalakturonases (ePGs) is SWAEe wat die homogalakturoniese dele van die plantselwand verteer en is noodsaaklike virulensie faktore vir ‘n aantal swamme, onder andere Botrytis cinerea. ‘n Belangrike weerstandsmeganisme van plante behels die inhibering van swam SWAEe om sodoende die patogeen-aanval te stop of te vertraag. Die poligalakturonase-inhiberende proteïne (PGIPs) van plante is selwand-geassosieerde SWAEinhiberende proteïne wat spesifiek teen swam ePGs optree. Verskeie verskillende PGIPs vanuit verskillende plantspesies is tot dusver beskryf. Dit is bekend dat hulle differensiële inhiberende vermoëns het wat dikwels toegeskryf kan word aan slegs ‘n paar belangrike aminosuurvolgordeverskille in die leusien-ryke herhalende (LRH) aktiewe areas. Die eerste wingerd PGIP is vantevore geïsoleer vanuit Vitis vinifera kultivar Pinotage (Vvpgip1) en gekarakteriseer. Hierdie Vvpgip1 geen is ooruitgedruk in die tabakspesie Nicotiana tabacum en was baie effektief om die weerstand van tabak teen die swam Botrytis cinerea te verhoog. Die ooruitdrukking van die transgeen, verhoogde PGIP aktiwiteit en goeie weerstand teen Botrytis cinerea is bevestig, en het gelei daartoe dat die transgeniese VvPGIP1 plantlyne geklassifiseer is as lyne met PGIP-spesifieke weerstandsfenotipes. ‘n Daaropvolgende transkriptomiese analise van die plantlyne het gewys dat hulle differensiële uitdrukking van selwand-geassosieerde gene het, asook biochemiese eienskappe, wat ‘n moontlike selwandversterking aandui in vergelyking met wilde-tipe tabak in die afwesigheid van infeksie. Die V. vinifera kultivars is hoogs vatbaar vir swamme, terwyl ander wingerdspesies, spesifiek die Noord-Amerikaanse spesies, bekend is vir hoë weerstand en selfs immuniteit teenoor verskeie patogeniese swamme. Sewe-en-dertig PGIPs is vantevore geïsoleer vanuit hierdie meer weerstandbiedende spesies. Die aminosuurvolgordes van die aktiewe areas van hierdie PGIPs is vantevore vergelyk met die van VvPGIP1 en dit is gevind dat hierdie proteïne hoogs homoloog is aan mekaar, sowel as aan VvPGIP1. Die verskillende nie-vinifera PGIPs het in 14 groepe verdeel na aanleiding van die homologie van hulle aktiewe areas. Vir hierdie studie is een PGIP vanuit elkeen van hierdie groepe gekies vir verdere funksionele analise in tabak. Die 14 nie-vinifera PGIP-koderende gene is stabiel oorgedra na tabak deur middel van Agrobacterium tumefaciens. Die vermeende transgeniese plante is geanaliseer vir die teenwoordigheid van die transgeen, die uitdrukking daarvan en PGIP aktiwiteit: bevestigde transgeniese tabak populasies wat wissel van drie tot 12 individuele getransformeerde lyne kon verkry word vir nege van die verskillende nie-vinifera PGIPs. ‘n Aantal van die transgeniese lyne is geïnfekteer met B. cinerea in twee onafhanklike heelplantinfeksies vir 11-14 dae om die siekteweerstand van hierdie PGIPs teenoor die swam te evalueer. ‘n Plantlyn van die VvPGIP1-populasie is as ‘n verwysing ingesluit en al die infeksies is vergelyk met die wilde-tipe tabak. Al die geïnfekteerde lyne wat die nie-vinifera PGIPs ooruitdruk het ‘n baie sterk afname in siektesimptome getoon in vergelyking met die wilde-tipe kontrole: na aanvanklikle primêre lesies gevorm het, is die verspreiding van die infeksie ingeperk en gestop in hierdie lyne, terwyl die wilde-tipe plante baie erg geaffekteer is. Terwyl die VvPGIP1 lyn ook die tipiese PGIPweerstandsrespons getoon het, het die nie-vinifera PGIPe kleiner lesies ontwikkel, wat dui op baie sterk weerstandsfenotipes. Die gekarakteriseerde nie-vinifera PGIP ooruitdrukkende lyne, asook die VvPGIP1 lyn en die wilde-tipe kontrole, is gebruik om die vorige waarneming dat die ooruitdrukking kan lei tot veranderinge in selwandgeen-uitdrukking verder te ondersoek. Analise van die uitdrukking van ‘n xiloglukaan-endotransglikosilase (xth) geen in die transgeniese populasie het getoon dat hierdie geen afgereguleer is in gesonde, oninfekteerde weefsel van al die transgeniese lyne wat getoets is. Dit het vorige resultate bevestig en het ook bevestig dat hierdie geen afgereguleer is in alle wingerd PGIP-ooruitdrukkende lyne wat tot dusver getoets is. XTH is tipies betrokke by selwandmetabolisme, spesifiek by die beheer van selwandsterkte en selwandelastisiteit. Dit is uit vorige werk bekend dat die afregulering van hierdie geen lei tot versterking van die plantselwand. Die resultate verkry tydens hierdie studie het gewys dat die PGIP-spesifieke weerstand fenotipe van VvPGIP1-ooruitdrukkende tabak ook bevestig kon word in transgeniese tabak wat nie-vinifera PGIPs vanuit meer weerstandbiedende wingerdspesies ooruitdruk. Die feit dat hierdie PGIP lyne almal selfs beter weerstand teen B. cinerea bied as VvPGIP1 lyne is ‘n interessante invalshoek vir opvolgende ondersoeke na die belang van strukturele verskille tussen die nie-vinifera PGIPs en VvPGIP1. Hierdie transgeniese lyne is ook uitstekende hulpbronne om die in vivo funksies van PGIPs verder te bestudeer in die konteks van plantpatogeen interaksies.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosach, 2010. / ENGLISH ABSTRACT: Cabernet Sauvignon, the most planted red wine cultivar in South Africa, is prone to vigorous growth with low yields. The aim of the study was to describe how Cabernet Sauvignon grapevines react to climate and irrigation within the Swartland region. Such knowledge would assist growers in decisions regarding long term as well as short term cultivation practices. This study was part of a larger project carried out by the Infruitec-Nietvoorbij institute of the Agricultural Research Council at Stellenbosch to determine effects of soil type and climate on yield and wine quality of Cabernet Sauvignon. The larger project was carried out in selected grape growing regions, i.e. Stellenbosch, Swartland, Lower Olifants River and Lower Orange River. Due to the proximity to the Atlantic Ocean, the study area in the Swartland region could be divided into two climatic regions for viticulture. Grapevines near Philadelphia closer to the ocean experienced less water constraints compared to those further inland near Wellington. Variation in stem water potential could also be related to soil water matric potential. Climate tended to have a more pronounced effect on the grapevine response to water constraints further inland than closer to the ocean. Vegetative growth, berry size and yield depended on water constraints experienced by the grapevines. In the warmer climate, severe constraints reduced yield. In the warmer climate, grapes started to ripen earlier than those in the cooler climate. Sugar concentration (mg/mL) was highest where grapevines experienced moderate water constraints. These seemingly balanced grapevines had the highest sugar accumulation, probably due to optimum photosynthesis and carbohydrate utilization. Low water constraints increased vegetative growth which could have been a sink for sugar loading. In addition to sugar loading, degree Balling (°B) increases could also have been due to a concentration effect where water constraints reduced berry volume. Therefore, °B is probably not a representative indicator of grapevine functioning. Anthocyanin biosynthesis, as quantified on a per berry basis, showed that sugar and anthocyanin could be co-regulated, with anthocyanin biosynthesis reaching a plateau when the sugar content per berry reached 200 mg/mL to 220 mg/mL. At véraison, the most intense grape colour occurred where grapevines experienced moderate water constraints, i.e. single drip line at Wellington and no irrigation at Philadelphia. However, at harvest grapes from the cooler climate tended to have more intense colour and higher phenolics, indicating that lower temperatures favoured anthocyanin biosynthesis. These results supported earlier findings that grapevine water status influences berry volume and dynamics of berry ripening. Water constraints tended to increase sensorial wine colour intensity, as well as wine fullness. Moderate water constraints at both localities resulted in the best sensorial wine quality. Yet there were indications that too severe water constraints could be detrimental to wine quality. Irrigation can be used to manipulate grapevine growth in warmer climates, but might be less effective in cooler climates. In warmer climates, moderate water constraints required to achieve balanced grapevine functioning can be obtained with single drip irrigation, but this might not be the case in cooler climates. / AFRIKAANSE OPSOMMING: Cabernet Sauvignon, die mees aangeplante rooiwynkultivar in Suid-Afrika, is geneig tot kragtige groei met lae opbrengste. Die doelwit van hierdie studie was om te beskryf hoe Cabernet Sauvignon-wingerdstokke reageer op klimaat en besproeiing in die Swartland-streek. Hierdie kennis sal kwekers help wanneer hulle besluite moet neem oor langtermyn sowel as korttermyn verbouingspraktyke. Hierdie studie was deel van ‘n groter projek deur die Infruitec-Nietvoorbij Instituut van die Landbounavorsingsraad op Stellenbosch om die effekte van grondtipe en klimaat op die opbrengs en wynkwaliteit van Cabernet Sauvignon te bepaal. Die groter projek is in geselekteerde wingerdverbouingstreke uitgevoer, nl. Stellenbosch, Swartland, Benede Olifantsrivier en Benede Oranjerivier. As gevolg van die nabyheid daarvan aan die Atlantiese Oseaan kon die studiegebied in die Swartland-streek in twee klimaatstreke vir wingerdbou verdeel word. Wingerdstokke by Philadelphia, wat nader aan die oseaan is, het minder waterstremming ervaar as dié verder binnelands naby Wellington. Veranderinge in stamwaterpotensiaal hou moontlik ook verband met die grondwater- matrikspotensiaal. Klimaat het ‘n groter effek op die reaksie van die wingerdstok op waterstremming verder binnelands as nader aan die oseaan. Vegetatiewe groei, korrelgrootte en opbrengs was afhanklik van die waterstremminge wat deur die wingerdstokke ervaar is. In die warmer klimaat het die ernstige stremminge opbrengs verminder. In die warmer klimaat begin druiwe vroeër ryp word as in die koeler klimaat. Suikerkonsentrasie (mg/ml) was die hoogste waar wingerde matige waterstremming ervaar het. Hierdie skynbaar gebalanseerde wingerdstokke het die hoogste suikerakkumulasie vertoon, moontlik as gevolg van optimum fotosintese en koolhidraatverbruik. Lae waterstremming het vegetatiewe groei verhoog, wat ook ‘n vraagpunt vir suikerlading kon wees. Benewens suikerlading kon verhogings in grade Balling (°B) ook moontlik aan ‘n konsentrasie-effek toegeskryf word in terme waarvan waterstremming die korrelvolume verminder het. °B is dus moontlik nie ‘n verteenwoordigende indikator van wingerdstokfunksionering nie. Antosianienbiosintese, soos gekwantifiseer op ‘n per-korrel basis, het getoon dat suiker en antosianien saam gereguleer kon word, en dat antosianienbiosintese ‘n plato bereik het wanneer die suikerinhoud per korrel 200 mg/mL tot 220 mg/mL was. By deurslaan het die mees intense druifkleur voorgekom waar die wingerdstokke matige waterstremming ervaar het, d.w.s. enkel druplyn op Wellington en geen besproeiing op Philadelphia. Teen oes was die druiwe in die koeler klimate egter geneig om meer intense kleur en meer fenole te bevat, wat aandui dat laer temperature antosianienbiosintese bevoordeel. Hierdie resultate ondersteun vroeër bevindings dat die waterstatus van die wingerdstok ‘n invloed op korrelvolume en die dinamika van korrelrypwording het. Waterstremming neig om die sensoriese wynkleurintensiteit te verhoog, asook die volheid van die wyn. Matige waterstremming op beide liggings het aanleiding gegee tot die beste sensoriese wynkwaliteit. Tog was daar aanduidings dat waterstremming wat te straf was, nadelig kon wees vir wynkwaliteit. Besproeiing kan gebruik word om wingerdgroei in warmer klimate te manipuleer, maar is moontlik minder effektief in koeler klimate. In warmer klimate kan die matige waterstremming wat benodig is vir gebalanseerde wingerdstokfunksionering, verkry word deur enkel drupbesproeiing, maar dit is moontlik nie die geval in koeler klimate nie.

Thesis (MScAgric)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: In recent years South African wines have been under the spotlight due to references in the international wine media to a distinctive ‘burnt rubber’ character. Many winemakers and wine experts argued that the peculiar character could be ascribed to winemaking errors linked to mismanaged fermentation. An alternative possible source of the taint was identified in the coal tar creosote used as a wood preservative in vineyard trellis systems. South African regulations allow for the use of creosoted utility poles in agricultural land, but in Europe and USA their usage has been highly regulated and even banned for certain applications. Contamination of foodstuff by polycyclic aromatic hydrocarbons (PAHs) is one of the major motives for the banning of creosote in Europe and USA. Some of the compounds in the creosote mixture display very strong odour characteristics and for this reason it became the focus of attention for the present study. The overall aim of this study was to determine if vines trellised with creosoted posts could accumulate or absorb the various malodorous compounds deriving from the wood treatment onto the grape berries. These compounds could then be extracted from the grape berries into the wine during alcoholic fermentation, creating quality and sensory problems. Chapter 2 of this thesis gives an overview of the extensive literature that deals with methods of analysis of PAHs and malodorous phenols using both Gas Chromatography (GC), as well as High Performance Liquid Chromatography (HPLC). New methods for sample preparation, as well as analysis of PAHs using HPLC-Diode Array Detector (DAD), were developed and the results reported in Chapter 3. It was demonstrated that Sauvignon blanc experimental wines contained only chrysene at very low levels. The concentrations of chrysene found in the experimental wines are within the prescribed parameters as established by The Commission of the European Communities. Since no other PAH compounds were found in the samples analysed, it was concluded that the experimental wines were safe for human consumption and complied with EU regulations. The effect of vineyard posts on the sensorial characteristics of wine is discussed in Chapter 4. Creosoted poles were found to be responsible for an off-flavour described as ‘burnt rubber’ and ‘tarry’ in Merlot wines produced from grapes grown in close proximity to the posts. Following some of the reported findings, new guidelines have been introduced in the Integrated Production of Wine certification, which advise against the use of creosoted poles for vineyard trellising. This preliminary but important guideline will bring the South African wine industry a step closer to the fulfilling the obligations for food safety as required by the legislation of our major export partners. Future investigations are recommended to completely understand and evaluate the cumulative effect of creosoted posts in a fully trellised vineyard. / AFRIKAANSE OPSOMMING: Oor die afgelope paar jaar is Suid-Afrikaanse wyne onder die soeklig geplaas as gevolg van verwysings in die internasionale wynmedia na ‘n duidelike ‘gebrande rubber’-karakter. Baie wynmakers en wyndeskundiges het aangedui dat hierdie besonderse karakter toegeskryf kan word aan wynbereidingsfoute wat verband hou met gisting wat wanbestuur is. ‘n Alternatiewe moontlike oorsprong van die smaak is geïdentifiseer in die koolteer wat as ‘n houtpreserveermiddel in wingerdopleistelsels gebruik word. Suid-Afrikaanse regulasies maak voorsiening vir die gebruik van kreosoteerde nutspale in landbougrond, hoewel hulle gebruik in Europa en die VSA hoogs gereguleerd en in sommige gevalle selfs verbied is. Die besmetting van kossoorte deur polisikliese aromatiese koolwaterstowwe (polycyclic aromatic hydrocarbons (PAHs)) is een van die vernaamste redes vir die verbanning van kreosoot in Europa en die VSA. Sommige van die verbindings in die kreosootmengsel het baie sterk geurkenmerke en daarom is dit die fokus van die huidige studie. Die oorhoofse doelwit van hierdie studie was om te bepaal of wingerde wat op kreosoteerde pale opgelei is, die verskillende onwelriekende verbindings afkomstig van die houtbehandeling in die druiwekorrels kan akkumuleer of absorbeer. Hierdie verbindings sou dan tydens alkoholiese gisting uit die druiwekorrels in die wyn geëkstraheer kon word, wat aanleiding sou gee tot kwaliteits- en sensoriese probleme. Hoofstuk 2 van hierdie tesis verskaf ‘n oorsig van die breedvoerige literatuur wat handel oor metodes om PAH’s en onwelriekende fenole met behulp van beide gaschromatografie (GC) en hoëdrukvloeistofchromatografie (HPLC) te analiseer. Nuwe metodes is ontwikkel om monsters voor te berei en om PAH’s met behulp van ‘n HPLC-diode array detector (DAD) te analiseer. Die resultate word in Hoofstuk 3 gerapporteer. Daar is aangetoon dat die eksperimentele Sauvignon blanc-wyne chriseen teen baie lae vlakke bevat het. Die konsentrasies van chriseen wat in die eksperimentele wyne gevind is, is binne die voorgeskrewe parameters van die Kommissie van die Europese Gemeenskap. Aangesien daar nie ander PAHverbindings in die geanaliseerde monsters gevind kon word nie, is daar tot die gevolgtrekking gekom dat die eksperimentele wyne veilig is vir menslike verbruik en aan die EG-regulasies voldoen. Die effek van wingerdpale op die sensoriese kenmerke van wyn word in Hoofstuk 4 bespreek. Kreosoteerde pale is gevind wat verantwoordelik is vir ‘n wangeur in Merlot-wyne afkomstig van druiwe wat naby die pale gegroei het en wat as ‘gebrande rubber’ en ‘teeragtig’ beskryf is. Op grond van sommige van die gerapporteerde bevindings, is nuwe riglyne ingesluit in die sertifisering vir die Geïntegreerde Produksie van Wyn, wat aanbeveel dat kreosoteerde pale nie vir wingerdoplei gebruik word nie. Hierdie voorlopige, maar belangrike riglyn sal die Suid-Afrikaanse wynbedryf al beter in staat stel om te voldoen aan die voedselveiligheid regulasies wat vereis word deur die wetgewing van ons belangrikste uitvoervennote. Toekomstige ondersoeke moet aangewend word om die kumulatiewe effek van kreosoteerde pale in volledig opgeleide wingerde ten volle te verstaan en te evalueer. / Indaba Scholarship and Winetech for their financial support

Dissertation (PhD)--University of Stellenbosch, 2003. / ENGLISH ABSTRACT: A survey was conducted in the Western Cape Province during the 1999/2000 and 2000/2001 seasons on mealybugs occurring in vineyards. P/anococcus ficus (Signoret) was the dominant mealybug in vineyards during this time. During this study P. ficus was recorded for the first time on roots of grapevines, which has far reaching implications for the control of this important vine leafroll virus vector as control actions were focused on above ground control. Other mealybugs presently recorded in local vineyards included Pseudococcus /ongispinus (Targioni) and Ferrisia ma/vastra (McDaniel). Pseudococcus viburni (Maskell) and Ps. so/ani Ferris were found on weeds in vineyards. Natural enemies of P. ficus recorded most frequently were species of Nephus predatory beetles, and the parasitaids Coccidoxenoides peregrinus (Timberlake), Anagyrus sp. and Leptomastix dacty/opii (Howard). Developmental studies on P. ficus and C. peregrinus indicated that the intrinsic rate of increase (rm) was similar, peaking at 25°C (rm = 0.169 for P. ficus; rm = 0.149 for C. peregrinus). The net replacement rate (Ra) was higher for P. ficus than for C. peregrinus at all five temperatures tested. The Ra for P. ficus reached a maximum at 21°C (308.87) and C. peregrinus at 25°C for C. peregrinus (69.94). The lower and upper thresholds for development of P. ficus were estimated at 16.59 and 35.61°C respectively. The lower threshold for development of C. peregrinus was 8.85°C. These parameters indicated that both insects were well adapted to temperatures in the Western Cape Province. The lower minimum threshold temperature of C. peregrinus in relation to that of P. ficus suggests that C. peregrinus should be more active during winter and early spring than P. ficus. A central systematic presence-absence sampling system was developed for P. ficus. Monitoring three different plant parts on the vine indicated that new growth areas on vines adjacent to the main stem could serve as an early warning system for pending P. ficus bunch infestations. Intervention should be planned when 2 % of the stems are infested with P. ficus when using this system. Seasonal population studies of P. ficus and its natural enemies showed that stem infestation by P. ficus reached peak levels during January in Robertson and Stellenbosch and during February in the Hex River Valley. Vine mealybugs colonised new growth early in the season, followed by the leaves and eventually the bunches towards the end of the season. High stem infestations early in the season resulted in high bunch infestation levels at harvest. A density dependent relationship was evident between P. ficus populations and parasitoid populations, suggesting that the parasitoids played a mayor role in the biological control of P. ficus populations. Biological control was however only achieved towards the end of the season when damage to the crop had already occurred. Mass releases of C. peregrinus on P. ficus populations were done in order to augment biological control as an alternative to chemical control. Between five and six releases of 20 000 C. peregrinus per release were done at monthly intervals in three grapegrowing areas. Mass released C. peregrinus controlled P. ficus adequately in the Hex River Valley. Control of P. ficus using this approach was no worse than using chemical control in Robertson and Stellenbosch. C. peregrinus is commercially available and can therefore be used as an alternative to chemical control by producers. Degree day estimation was used to predict development of P. ficus populations. This information was used as an input in a P. ficus pest management model. Data acquired from P. ficus and ant monitoring were used as components to construct a decision chart. This chart can be used by producers to optimise the control of P. ficus populations using either chemical control or mass releases of C. peregrinus. / AFRIKAANSE OPSOMMING: "n Studie is gedurende die 1999/2000 en 2000/2001 seisoene gedoen met die doelom die witluisspesies wat in wingerde voorkom, te identifiseer. Planococcus ficus (Signoret) is tans die dominante witluisspesie in wingerde in die Wes Kaap Provinsie. P. ficus kolonies is op wingerdwortels gevind. Dié bevinding kan verreikende gevolge hê vir die beheer van dié plaag as "n belangrike rolbladvirus vektor aangesien beheer tot dusver gefokus het op bogrondse gedeeltes. Ander witluisspesies wat in wingerde gevind is, sluit in Pseudococcus /ongispinus (Targioni) en Ferrisia malvastra (McDaniel). Pseudococcus vibumi (Maskell) en Ps. so/ani Ferris is op onkruide in wingerde gevind. Dominante natuurlike vyande van P. ficus sluit predatoriese kewertjies van verskeie Nephus spp. en die parasitoïede Coccidoxenoides peregrinus (Timberlake), Anagyrus sp. en Leptomastix dacty/opii (Howard) in. Ontwikkelingstudies op P. ficus en C. peregrinus het aangetoon dat die inhirente voortplantingstempo (rm) soortgelyk was vir beide insekte met "n maksimum by 25°C (0.169 vir P. ficus, 0.149 vir C. peregrinus). Die netto vervangingstempo (Ra) was in vergelyking met C. peregrinus hoër vir P. ficus by al vyf temperature getoets. Die Ra van P. ficus het "n maksimum bereik teen 21°C (308.87) en die van e. peregrinus by 25°C (69.94). Die teoretiese hoër en laer drempels vir ontwikkeling van P. ficus was onderskeidelik 16.59 en 35.61 oe. Die teoretiese laer drempelwaarde van ontwikkeling vir e. peregrinus was 8.85°e. Hierdie parameters dui aan dat beide insekte goed aangepas is by temperature in die Wes Kaap Provinsie. Die laer minimum drempel vir ontwikkeling van C. peregrinus in verhouding tot P. ficus impliseer dat C. peregrinus in die winter en vroeë lente meer aktief sal wees as P. ficus. 'n Sentrale sistematiese aan-afwesig moniteringsisteem met bekende vlakke van steekproefnemingsfout is ontwikkel in kommersiële wingerde vir P. ficus. Monitering van drie verskillende dele op die wingerdstok het aangedui dat die nuwe groei areas kan dien as 'n vroeë waarskuwing vir latere P. ficus trosinfestasies. Dié sisteem sal produsente in staat stelom te bepaal wanneer optrede noodsaaklik is. Daar word voorgestel dat optrede noodsaaklik is by 'n P. ficus besmettingsvlak van 2 % op die nuwe groei areas op stokke. Stambesmetting deur P. ficus het in Januarie piekvlakke bereik in Stellenbosch en Robertson, en in Februarie in die Hex Rivier Vallei. P. ficus koloniseer nuwe groei vroeg in die seisoen waarna blare en trosse aan die einde van die seisoen gekoloniseer word. Dié data dui aan dat P. ficus besmetting op nuwe groei vroeg in die seisoen 'n aanduiding kan gee van hoë trosbesmetting aan die einde van die seisoen. 'n Digtheidsafhanklike verwantskap was waarneembaar tussen P. ficus plaagpopulasies en parasitoïed populasies. Dié verwantskap dui aan dat parasitoïede die belangrikste rol speel in biologiese beheer van P. ficus populasies. Biologiese beheer van witluis is egter eers aan die einde van die seisoen bereik toe die oes reeds beskadig was. Massavrylatings van C. peregrinus is in P. ficus besmette blokke gedoen om biologiese beheer aan te help en sodoende as alternatief tot chemiese beheer te dien. Tussen vyf en ses vrylatings met 20 000 C. peregrinus is een keer per maand gedurende die seisoen gedoen. Die vrygelate C. peregrinus het P. ficus populasies voldoende beheer in die Hex Rivier Vallei. Beheer van P. ficus deur massavrylatings van C. peregrinus was soortgelyk as chemiese beheer in Robertson en Stellenbosch. C. peregrinus is kommersieel beskikbaar en kan om hierdie rede as alternatief tot chemiese beheer gebruik word. Graaddag bepaling is gebruik om die ontwikkeling van P. ficus populasies te voorspel. Hierdie inligting is gebruik as 'n verdere hulpmiddel in die P. ficus plaagbeheermodel. Inligting verkry vanuit P. ficus en mier monitering is gebruik as komponente in die opstel van 'n besluitnemingstabel. Hierdie tabel kan gebruik word deur produsente om beheer van P. ficus plaagpopulasies te optimaliseer deur chemiese beheer of massavrylatings van C. peregrinus.

Thesis (MSc)--Stellenbosch University, 2003. / ENGLISH ABSTRACT: Paecilomyces Ii/acinus, ras 251 (geregistreer in terme van wet 36 van 1947 as Suid-Afrika se eerste natuurlike nematisiede en kommersieel beskikbaar as PI Plus) is as biologiese beheer agent getoets by aartappels en in geïntegreerde beheer programme by sitrus en wingerd teen respektiewelik Me/oidogyne species, Ty/enchu/us semipenetrans en verskeie ektoparasitiese nematodes. Die swam toon belofte vir die beheer van hierdie nematodes en het terselfdertyd nie 'n nadelige effek op nie-teiken, voordelige organismes in die grond nie. Veral in kombinasie met chemiese middels, as deel van geïntegreerde programme, kan dit lei tot verminderde gebruik van hoogs toksiese middels en dus meer omgewingsvriendelike landboupraktyke. Biological control of plant parasitic nematodes on potatoes, citrus and grapevine with the fungus, Paecilomyces liIacinus. Paecilomyces liIacinus, race 251 (registered in terms of act 36 of 1947 as South Africa's first natural nematicide, commercially available as PI Plus) was tested as a biological control agent on potatoes and in integrated control programs on citrus and grapevine against Me/oidogyne species, Ty/enchu/us semipenetrans and various ectoparasitic nematodes respectively. The fungus shows promise for the control of these nematodes, without having a harmful effect on non-target, beneficial organisms in the soil. Especially in combination with chemical products, as part of integrated programs, this can lead to less use of highly toxic compounds and thus to more environmentally friendly agricultural practices. / AFRIKAANSE OPSOMMING: Sedert die ontdekking van die swam, Paeci/omyces Ii/acinus (Thom.) Samson as 'n effektiewe eierparasiet van Meloidogyne incognita acrita en Globodera pal/ida (Jatala et al., 1979) het verdere veldproewe in Peru tot die effektiewe beheer van M. incognita en Tylenchulus semipenetrans gelei. Na verskeie suksesse in Peru is die swam onder verskillende klimaat- en grondkondisies in verskeie ander lande beproef. Die sukses behaal in die Filippyne het gelei tot die kommersiële produksie van die swam onder die handelsnaam Biocon. Anders as met chemiese middels vind die werking van biologiese agente stadig oor tyd plaas. Biologiese beheer sal nie chemiese beheer sonder meer kan vervang nie. Dit behoort egter deel te vorm van geïntegreerde nematode bestuur. Inkorporering van die natuurlike organismes, die oordeelkundige gebruik van chemiese nematisiedes, moontlik in kombinasie met die biobeheer agente, weerstand, en ander kulturele praktyke moet ernstig oorweeg word as ons hoop om die steeds groeiende wêreldbevolking te voed (Jatala, 1986). Paecilomyces liIacinus, ras 251, Suid-Afrika se eerste geregistreerde natuurlike nematisiede, kommersieel beskikbaar as PI Plus, is in die Olifantsrivier besproeiingsgebied geëvalueer vir die bestuur van ekonomies belangrike plantparasitiese nematodes by aartappels, sitrus en wingerd. Hierdie gewasse is belangrike bedryfstakke van die streek en is onderhewig aan skade deur nematodes wat die opbrengs nadelig beïnvloed. Chemiese beheer bied slegs 'n korttermyn oplossing vir nematode probleme en skadelike getalle word in 'n kort tyd weer opgebou. Boonop lei dié hoogs toksiese middels tot agteruitgang van die omgewing en sy waterbronne. Die toenemende besorgdheid hieroor en die groot potensiaal van biologiese beheer agente (Jatala, 1986) was die hoofrede vir die werk waaroor hier gerapporteer word.

Heat induced protein haze is a common problem in white wine. Grape derived pathogenesis related proteins slowly denature and aggregate during wine storage and this gives rise to light dispersing haze. Protein haze formation is currently prevented by removing proteins using bentonite, an aluminium silicate clay, but this method has drawbacks. A potential alternative or complementary method is the use of haze protective factors ( HPF ), specific mannoproteins from Saccharomyces cerevisiae that visually reduce protein haze. Hpf1p was originally isolated from Muscat Gordo Blanco wine and Hpf2p from a synthetic grape juice ferment. Based on partial amino acid sequences, putative structural genes, HPF1 and HPF2, for these proteins were identified. HPF1 has a homologue, HPF1 ', ( 71 % similarity ) in S. cerevisiae. Sequence analysis suggests that Hpf1p, Hpf1 ' p and Hpf2p are localised to the cell wall or plasma membrane. This study aimed to determine the biological function of the HPF genes in S. cerevisiae. HPF overexpression and deletion strains were constructed and analysed for cell wall related phenotypes. Under a number of conditions, including cold temperature and ethanol stress, the hpf1 Δ hpf1 ' Δ strain was more tolerant than the wild type strain. However, mating efficiency of the hpf1 Δ hpf1 ' Δ strain was significantly less than the wild type strain and this was found to be correlated with the persistence of a septum between the mating partners. The decreased mating efficiency was also mating type specific, only occurring in MAT α cells. This study also aimed to establish conclusively that the HPF genes do indeed encode proteins with haze protective properties. Haze protective activity of the material from ferment supernatants was assessed. Material from the HPF deletion strains exhibited significantly less haze protective activity than the wild type. Moreover, material derived from HPF1 and HPF1 ' overexpressors was more active than material from the wild type. A 6xHis - tagged Hpf2p was expressed and purified using immobilised metal affinity chromatography. This Hpf2p had significant haze protective activity. Modification of N - glycans of 6xHis - Hpf2p by Endoglycosidase H decreased its haze protective activity.visually reduce protein haze. Hpf1p was originally isolated from Muscat Gordo Blanco wine and Hpf2p from a synthetic grape juice ferment. Based on partial amino acid sequences, putative structural genes, HPF1 and HPF2, for these proteins were identified. HPF1 has a homologue, HPF1 ', ( 71 % similarity ) in S. cerevisiae. Sequence analysis suggests that Hpf1p, Hpf1 ' p and Hpf2p are localised to the cell wall or plasma membrane. This study aimed to determine the biological function of the HPF genes in S. cerevisiae. HPF overexpression and deletion strains were constructed and analysed for cell wall related phenotypes. Under a number of conditions, including cold temperature and ethanol stress, the hpf1 Δ hpf1 ' Δ strain was more tolerant than the wild type strain. However, mating efficiency of the hpf1 Δ hpf1 ' Δ strain was significantly less than the wild type strain and this was found to be correlated with the persistence of a septum between the mating partners. The decreased mating efficiency was also mating type specific, only occurring in MAT α cells. This study also aimed to establish conclusively that the HPF genes do indeed encode proteins with haze protective properties. Haze protective activity of the material from ferment supernatants was assessed. Material from the HPF deletion strains exhibited significantly less haze protective activity than the wild type. Moreover, material derived from HPF1 and HPF1 ' overexpressors was more active than material from the wild type. A 6xHis - tagged Hpf2p was expressed and purified using immobilised metal affinity chromatography. This Hpf2p had significant haze protective activity. Modification of N - glycans of 6xHis - Hpf2p by Endoglycosidase H decreased its haze protective activity. / Thesis (Ph.D.)--School of Agriculture and Wine, 2003.

Disputats. Universität Hohenheim, 2002. / Haves kun i elektronisk udg. Thesis (Ph.D.). 41 ill., 5 tables; approx. 200 ref. Summaries (En, Ge).

This thesis presents results from two separate studies. First, the impact of bearer length on yield components within the canopy was investigated in season 2005/06, on a commercially-managed, mechanically-pruned vineyard of Vitis vinifera L. Cabernet Sauvignon in Coonawarra, South Australia. Pruning resulted in the retention of bearers with 1-7 nodes, with the weighted average bearer length being two nodes for the canopy. As bearers of one to five nodes in length were the most common, these were studied. Yield components (on a per shoot basis) were analysed according to the node position on the bearer at which the shoot arose. Both budburst and inflorescence number per node were highest at the distal node positions on each length bearer, even if the nodes were at the same positions from the base of the bearer and would normally be expected to have similar fertility. Budburst appeared to act by modifying inflorescence number per node based on the relative location of each node from the apex of the bearer. Shoots that arose from the most distal node positions had the highest flower number per inflorescence and berry number per bunch. Flower number per inflorescence was significantly higher on two-inflorescence shoots than single-inflorescence shoots. The relationship between bunch size and node position, unlike that between inflorescence number and node position, was dependent on bearer length. The relative size of the inflorescence appeared to be affected more so by the node pOSition at which the shoot occurred on the bearer, as opposed to the actual node position on the shoot at which the inflorescence occurred. There was a positive, non-linear relationship between average fruit yield per bearer and bearer length. Although yield was highest from the bearer with the highest node number (five nodes), there was no significant difference in yield per bearer for the bearers of three to five nodes in length. If average bearer length was increased from two to three nodes, the potential yield gain per bearer is estimated at 38 per cent. The second study presents results of correlations between bunch number and components of bunch weight (flower number and berry number) to investigate co-development of bunch number and bunch size. These data were collected from 4 vineyards in the Limestone Coast Zone of South Australia from Vilis vinifera L. Chardonnay, Shiraz and Cabemet Sauvignon during seasons 2002/03 to 2006/07. The significant correlations found between fertility and both bunch weight and flower number per inflorescence suggest that the same factors that affect bunch number in a particular season will also affect bunch size. When inflorescence primordia were initiated and differentiated under cool conditions, actual bunches per node and flowers per inflorescence were low. Differences in climate between the vineyard sites were found to be minimal and therefore did not strongly affect the magnitude of the yield components at the vineyard sites. Cultural practices at each vineyard site were sufficiently variable to affect fertility levels. Genotype is thought to determine the range of flowers per inflorescence that a variety can potentially carry, whereas actual flower number per inflorescence is thought to be determined by inflorescence primordium initiation and differentiation temperatures, as well as temperatures during budburst. Despite significant correlations between flower number per inflorescence and berry number per bunch, flower number per inflorescence preflowering for Cabemet Sauvignon, Shiraz and Chardonnay is inversely related to actual percentage fruit set. This is possibly a survival mechanism for the grapevine as it allows the vine to maximise yield each season without detriment to its longevity. Bunches per vine accounted for the majority of the seasonal variation in yield per vine. Fluctuations in bunch number per vine (and therefore yield) are likely to be reduced by varying the number of nodes retained per vine according to the relative fruitfulness per node present pre-pruning. This practice is therefore likely to result in the seasonal variation of berries per bunch becoming a stronger driver of yield. The commercial impacts of these studies are two-fold. Data presented will assist growers to understand the reasons for which their pruning regimes are affecting yield production and how these pruning regimes may be modified to achieve a target yield-particularly when growers are faced with seasons of low predicted fertility. In addition, data presented will allow growers to improve their crop forecasting accuracy, with a greater understanding of the link between bunch number and bunch size. In the current situation of oversupply in the wine industry, wineries are adopting a tough stance towards growers over-delivering on their grape contracts. Therefore, any assistance that can be provided to growers on improving accuracy of yield estimates will be beneficial both to the grower and winery. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1352744 / Thesis (M.Ag.Sc.) -- University of Adelaide, School of Agriculture, Food and Wine, 2009

This thesis presents results from two separate studies. First, the impact of bearer length on yield components within the canopy was investigated in season 2005/06, on a commercially-managed, mechanically-pruned vineyard of Vitis vinifera L. Cabernet Sauvignon in Coonawarra, South Australia. Pruning resulted in the retention of bearers with 1-7 nodes, with the weighted average bearer length being two nodes for the canopy. As bearers of one to five nodes in length were the most common, these were studied. Yield components (on a per shoot basis) were analysed according to the node position on the bearer at which the shoot arose. Both budburst and inflorescence number per node were highest at the distal node positions on each length bearer, even if the nodes were at the same positions from the base of the bearer and would normally be expected to have similar fertility. Budburst appeared to act by modifying inflorescence number per node based on the relative location of each node from the apex of the bearer. Shoots that arose from the most distal node positions had the highest flower number per inflorescence and berry number per bunch. Flower number per inflorescence was significantly higher on two-inflorescence shoots than single-inflorescence shoots. The relationship between bunch size and node position, unlike that between inflorescence number and node position, was dependent on bearer length. The relative size of the inflorescence appeared to be affected more so by the node pOSition at which the shoot occurred on the bearer, as opposed to the actual node position on the shoot at which the inflorescence occurred. There was a positive, non-linear relationship between average fruit yield per bearer and bearer length. Although yield was highest from the bearer with the highest node number (five nodes), there was no significant difference in yield per bearer for the bearers of three to five nodes in length. If average bearer length was increased from two to three nodes, the potential yield gain per bearer is estimated at 38 per cent. The second study presents results of correlations between bunch number and components of bunch weight (flower number and berry number) to investigate co-development of bunch number and bunch size. These data were collected from 4 vineyards in the Limestone Coast Zone of South Australia from Vilis vinifera L. Chardonnay, Shiraz and Cabemet Sauvignon during seasons 2002/03 to 2006/07. The significant correlations found between fertility and both bunch weight and flower number per inflorescence suggest that the same factors that affect bunch number in a particular season will also affect bunch size. When inflorescence primordia were initiated and differentiated under cool conditions, actual bunches per node and flowers per inflorescence were low. Differences in climate between the vineyard sites were found to be minimal and therefore did not strongly affect the magnitude of the yield components at the vineyard sites. Cultural practices at each vineyard site were sufficiently variable to affect fertility levels. Genotype is thought to determine the range of flowers per inflorescence that a variety can potentially carry, whereas actual flower number per inflorescence is thought to be determined by inflorescence primordium initiation and differentiation temperatures, as well as temperatures during budburst. Despite significant correlations between flower number per inflorescence and berry number per bunch, flower number per inflorescence preflowering for Cabemet Sauvignon, Shiraz and Chardonnay is inversely related to actual percentage fruit set. This is possibly a survival mechanism for the grapevine as it allows the vine to maximise yield each season without detriment to its longevity. Bunches per vine accounted for the majority of the seasonal variation in yield per vine. Fluctuations in bunch number per vine (and therefore yield) are likely to be reduced by varying the number of nodes retained per vine according to the relative fruitfulness per node present pre-pruning. This practice is therefore likely to result in the seasonal variation of berries per bunch becoming a stronger driver of yield. The commercial impacts of these studies are two-fold. Data presented will assist growers to understand the reasons for which their pruning regimes are affecting yield production and how these pruning regimes may be modified to achieve a target yield-particularly when growers are faced with seasons of low predicted fertility. In addition, data presented will allow growers to improve their crop forecasting accuracy, with a greater understanding of the link between bunch number and bunch size. In the current situation of oversupply in the wine industry, wineries are adopting a tough stance towards growers over-delivering on their grape contracts. Therefore, any assistance that can be provided to growers on improving accuracy of yield estimates will be beneficial both to the grower and winery. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1352744 / Thesis (M.Ag.Sc.) -- University of Adelaide, School of Agriculture, Food and Wine, 2009