Thesis (PhD(Agric))--Stellenbosch University, 2012. / ENGLISH ABSTRACT: A lack of defects is required for successful table grape marketing, which pre-suppose optimal vine performance, berry development and post-harvest quality. The supply of mineral nutrients affects vine development, physiology and berry quality. Despite a vast amount of research conducted over decades, there remain many unresolved issues regarding table grape vine nutrition to ensure optimal table grape quality and shelve-life. Unjustified fertilisation practices often include excessive applications of nitrogen (N), potassium (K) and calcium (Ca). A four-year field trial was therefore conducted on a sandy soil in the Paarl district of South Africa, using grafted on Ramsey, and trained to a gable trellis system. Nitrogen, potassium and calcium were applied, singular or in combination, at rates up to 300% the calculated annual nutritional requirement. The effect of these excessive applications on table grape performance under typical South African cultivation conditions was investigated for Vitis vinifera L. cv. Prime Seedless, a very early seedless table cultivar that is produced with minimum berry diameter of 18mm, with special reference to 1) vegetative growth, 2) expression of grapevine nutrient availability through foliar analyses, 3) berry nutrient accumulation patterns of this early cultivar, 4) manipulation of berry nutrient content through soil and bunch directed applications and 5) the effect of berry nutrient content on its quality. No definite vegetative growth responses (expressed as shoot length, leaf surface area and shoot mass) and leaf chlorophyll content differences were obtained for all the treatments. These results were obtained in a vineyard on a sandy soil where excessive N fertilisation caused a reduction of soil pH to detrimentally low levels and where the excessive N, K and Ca applications reduced mutual concentrations and that of Mg, in the soil. A lack of stimulation in vegetative growth may therefore be ascribed to the combined negative effect of these excessive applications on soil pH and vine nutrition. Although the N content of petioles was higher for treatments where N was applied, consistent significant increases in petiole N with N fertilisation were not observed. Petiole N concentration showed a decreasing trend throughout the season. Petiole K concentrations were significantly increased by the K fertilisation at all phenological stages. None of the K fertilisation treatments, however, succeeded to raise petiole K concentrations above the accepted maximum norms and petiole K concentration at a specific sampling stage varied significantly between the four seasons. A general decrease in petiole K concentration was found for all seasons. Calcium fertilisation did not increase soil Ca content, resulting in a lack of differences in petiole Ca concentrations between treatments. An increase in petiole Ca concentration towards harvest was obtained. Correlations between petiole nutrient concentration and berry mineral content at harvest were poor. The only way of knowing the mineral content of berries would seem to be by measuring it directly instead of deducing it from the results of leaf or petiole analyses. The dynamics of berry growth impacted on berry nutrient concentration. Early rapid berry growth, predominantly due to cell division and cell growth, was associated with the most rapid decreases in N, P and Ca concentration. Due to mobility of K and Mg in the plant, that exceeds other nutrients, the decrease in concentration of these two mineral elements was not as pronounced as that of the others. Nutrient accumulation was most rapid during the pre-véraison period, but only Ca showed a definite termination during the early ripening period. The continued inflow of N, P, K and Mg, albeit at slower rates immediately after véraison, should be taken into consideration when fertilisation is applied. As a table grape, total accumulation of each nutrient in Prime Seedless berries also far exceeded that of other cultivars studied thus far. A particular difference is that the berry flesh:skin ratio is much higher than that of previously studied cultivars, leading to higher levels of nutrient accumulation in the flesh. Slightly larger berry size was obtained for N applications and is ascribed to slight increases in early vegetative growth, allowing a better response to GA3 treatments. The use of GA3 for berry enlargement is also considered the reason why K fertilisation, resulting in increased berry K levels, did not affect berry size, as is often found for wine grapes. Higher available NO3 - in the soil on account of excessive N applications resulted in higher levels of berry N, despite sub-optimal soil pH regimes that were created by these treatments. Berry K concentration and content were increased by K fertilisation. Rapid vine K uptake and translocation to the berries seem to negate the reduced vine nutritional status as observed in petioles for situations of over-fertilisation with N. Berry Ca levels were not increased by Ca fertilisation or by bunch applied Ca. The rapid rates of berry growth, together with low rates of berry Ca uptake and Ca uptake that terminates at the onset of ripening, are assumed to be the main reasons for this result. Low levels of decay as well as a lack of consistently increased decay were obtained for N containing treatments. Nitrogen levels in the berries above which their susceptibility to fungal infection is increased, should be established. Information on specific N compounds that may lead to more susceptibility is required. Potentially increased berry browning on account of high rates of K fertilisation needs to be further investigated; indications that this may occur were observed. Neither soil applied Ca nor bunch applied Ca improved berry quality, although Ca treatments seemed to reduce decay during the only season that significant differences were obtained. The negative effect of excessive fertilisation on soil chemistry of sandy soils has again been highlighted by this study. This annuls the fertilisation, leading to inefficient fertilisation and a lack of the desired responses. As indicator of vine nutrient availability, petiole analysis, was proven unreliable and should be evaluated in parallel with soil analyses, taking seasonal variation into consideration. The danger of being only guided by published norms for leaf nutrient concentrations when establishing fertilisation practices has again been highlighted by this study. This research indicated that for a very early cultivar like Prime Seedless, nutrient accumulation dynamics can already start to change during the pre-véraison period in some seasons. This is due to different edaphic and climatic conditions as well as berry size, which leads to much higher flesh:skin ratios. Future research on table grapes would need to develop an understanding of the various factors and dynamics that determine berry nutrient concentration and accumulation of early ripening, large berry sized, seedless table grape cultivars. / AFRIKAANSE OPSOMMING: Suksesvolle bemarking van tafeldruiwe is ten nouste afhanklik van die beskikbaarheid van druiwe sonder defekte, wat ‘n direkte verband met optimale wingerdprestasie, korrelontwikkeling en na-oes kwaliteit inhou. Voorsiening van minerale voedingstowwe beïnvloed die stok se groei, fisiologie en korrelgehalte. Ten spyte van ‘n oorweldigende hoeveelheid navorsing wat oor dekades reeds gedoen is, is daar steeds onopgeloste kwessies aangaande bemesting van tafeldruiwe vir optimale druifgehalte en houvermoë. Die gevolg is onoordeelkundige bemestingspraktyke wat o.a. aanleiding gee tot oorbemesting met stikstof (N), kalium (K) en kalsium (Ca). ‘n Vier-jaar-lange veldproef is gevolglik op ‘n sandgrond in die Paarl distrik (Suid-Afrika) onderneem deur gebruik te maak van Vitis vinifera L. cv. Prime Seedless geënt op Ramsey en op ‘n dubbel-gewel prieelstelsel opgelei is. Stikstof, K en Ca is alleen, of in kombinasie, toegedien teen hoeveelhede gelykstaande aan 300% van die wingerd se jaarlikse behoefte. Die effek van hierdie oormatige toedienings op tafeldruif prestasie onder Suid-Afrikaanse verbouingstoestande is ondersoek, met spesiale verwysing na 1) vegetatiewe groei, 2) uitdrukking van voedingstofbeskikbaarheid deur blaarontledings, 3) die voedingstof akkumulasie patrone van korrels van hierdie vroeë kultivar, 4) manipulasie van korrel voedingstofinhoud deur grond en trosgerigte toedienings en 5) die effek van korrel voedingstofinhoud op kwaliteit. Die doel van die proef was om bemestinspraktyke van Prime Seedless, ‘n baie vroeë pitlose tafeldruifkultivar met ‘n minimum korrelgrootte van 18 mm, te verfyn. Deur die akkumulasie patrone van die druiwe uit te klaar is daar ook ondersoek ingestel of oestyd en na-oes gehalte deur oormatige toediening van voedingstowwe affekteer word. Geen duidelike verskille betreffende vegetatiewe groeireaksies (uitgedruk as lootlengte, blaaroppervlaktes en lootmassas) asook verskille in blaar chlorofilinhoud is vir die behandelings verkry nie. Hierdie resultate is verkry in ‘n wingerd op ‘n sandgrond, waar oormatige N-bemesting aanleiding gegee het tot grond pH verlagings tot die peil van nadelige vlakke. Verder het die oormatige N, K en Ca toedienings wederkerige verlagings in konsentrasies, asook op dié van Mg, in die grond teweeggebring. Die tekort aan vegetatiewe groeiresponse op die behandelings kon dus toegeskryf word aan ‘n gekombineerde effek van die oormatige toedienings op grond pH en voedingstofbalanse. Hoewel die N-inhoud van bladstele hoër was vir behandelings wat N toediening ingesluit het, was daar nie konstante toenames in die vlakke verkry nie. Bladskyf N-konsentrasie het afgeneem deur die loop van die groeiseisoen. Vir alle fenologiese stadiums was bladskyf Kkonsentrasies betekenisvol verhoog deur K-bemesting. Nie een van die Kbemestingsbehandelings het egter daarin geslaag om bladskyf K inhoud vir enige monstertyd bo die algemeen aanvaarde maksimum norms te lig nie. Verder het bladskyf K inhoud by ‘n spesifieke fenologiese stadium ook betekenisvol tussen seisoene verskil. Die K-inhoud van bladskywe het afgeneem met verloop van die seisoen. Kalsiumbemesting het nie die grond se Ca inhoud deurgans verhoog nie, wat dus die tekort aan verskille in Ca konsentrasies tussen die behandelings verklaar. ‘n Toename in Ca konsentrasie en korrel Ca inhoud is vanaf set tot oes waargeneem. Swak korrelasies tussen bladskywe se voedingstofinhoude en korrels se voedingstofinhoude is verkry. Die enigste manier waarop korrels se voedingstofinhoude dus afgelei kan word, blyk te wees deur direkte bepaling daarvan. Voedingstofinhoude van korrels is deur groeipatrone daarvan beïnvloed. Vroeë korrelgroei, hoofsaaklik a.g.v. seldeling en selgroei, het met die vinnigste afnametempo van N, P en Ca gepaard gegaan. As gevolg van die hoër beweeglikheid van K en Mg in die plant in vergelyking met ander voedingstowwe, was die afname in konsentrasie van hierdie twee elemente nie so groot soos vir die ander nie. Voedingstofakkumulasie was die vinnigste in die periode voor deurslaan. Slegs Ca het ‘n beeïndiging van opname aan die einde van hierdie periode getoon. Die voortgesette opname van N, P, K en Mg, alhoewel stadiger kort na deurslaan, moet in ag geneem word wanneer bemesting toegedien word. Vir hierdie kultivar het die totale opname van elke bemestingstof dié van die ander kultivars wat tot hede bestudeer is, ver oorskry. ‘n Spesifieke verskil is ‘n baie hoër vleis:dop verhouding as wat vir ander kultivars verkry is. Dit gee aanleiding tot baie hoër vlakke van voedingstofakkumulasie in die vleis. Effens groter korrelgroottes is verkry waar N toedienings gemaak is. Dit word toegeskryf aan klein toenames in vroeë vegetatiewe groei, wat dus beter reaksie op GA3 behandelings tot gevolg gehad het. Die gebruik van GA3 vir korrelvergroting word ook beskou as die rede waarom K-bemesting, wat tot hoër vlakke van K in die korrels aanleiding gegee het, nie korrelgrootte, soos by wyndruiwe, bevorder het nie. Hoër NO3 - in die grond (water), na aanleiding van N toedienings, het aanleiding gegee tot hoer vlakke van N in die korrels. Dit het plaasgevind ten spyte van sub-optimale grond pH wat deur die oormatige N toedienings veroorsaak is. Korrel K konsentrasie en -inhoud is deur K-bemesting verhoog. Vinnige opname en translokasie van K na die korrels het ook geblyk die rede te wees waarom die verlaagde voedingstatus van die stokke a.g.v. oorbemesting met N nie die korrels se K inhoud geaffekteer het nie. Die vinnige groeitempo van die korrels, tesame met lae vlakke van Ca opname, asook korrels se Ca opname wat tydens rypwording ophou, word as die redes vir die tekorte aan behandelingseffekte beskou. Lae vlakke van bederf, asook ‘n tekort aan betroubare tendense dat bederf deur Nbemesting verhoog word, is verkry. Daar moet vasgestel word of daar N vlakke in die korrels is waarbo hul vatbaarheid vir swaminfeksies verhoog word, en of daar spesifieke N verbindings is wat die korrels meer vatbaar maak vir bederf. Indikasies dat K-bemesting interne verbruiningsvlakke verhoog het, regverdig verdere ondersoek. Korrelkwaliteit is nie deur grond- of trosgerigte toedienings bevoordeel nie. Die negatiewe effek van oormatige bemesting op die chemiese samestelling van sandgronde is weer deur hierdie navorsing uitgelig. Dit lei tot oneffektiewe bemesting en ‘n tekort aan die verlangde effekte. Blaarontledings blyk onbetroubaar te wees as aanduiding van voedingstof beskikbaarheid. Dit moet evalueer word saam met grondontledings en ook seisoenale variasie in ag neem. Die gevaar om slegs deur gepubliseerde norme gelei te word wanneer bemestingspraktyke bepaal word, is weer deur hierdie navorsing uitgelig. Voorst is daar in hierdie navorsing gevind dat voedingstof akkumulasiepatrone van ‘n baie vroeë kultivar soos Prime Seedless alreeds voor deurslaan begin verander a.g.v. omgewingstoestande en korrelgroei wat tot ‘n veel hoër vleis:dop verhouding aanleiding gee. Toekomstige navorsing op tafeldruiwe behoort die faktore en dinamika wat voedingstofkonsentrasie en -akkumulasie in korrels van vroeë, groot korrel, pitlose tafeldruifkultivars beïnvloed verder te ondersoek.

Thesis (MScAgric)--University of Stellenbosch, 2004. / ENGLISH ABSTRACT: The production of quality wine is influenced by numerous factors of which grape quality is one of the most important factors. The production of quality wine, however, is not possible without good winemaking techniques and effective quality control. Critical control points (CCP) during the winemaking process must be identified to ensure optimum wine quality. Grape must is a complex medium that contains different micro-organisms which can be either beneficial or negative to wine quality, depending on the physical and chemical conditions that prevail in the must. Yeasts are responsible for alcoholic fermentation, lactic acid bacteria (LAB) for malolactic fermentation (MLF) and acetic acid bacteria (AAB) for the production acetic acid from ethanol. Yeasts and certain LAB can also produce acetic acid and thereby increasing the volatile acidity (VA) of wine. These micro-organisms can influence each other in complex fashions by competing for growth nutrients and by producing inhibitory substances. Most winemakers nowadays use commercial yeast strains to inoculate wine fermentations. This, however, does not assure a problem-free fermentation and cases of stuck and sluggish fermentations are annually reported worldwide. In these or most cases fermentation takes longer than 21 days to complete and the wine contains a residual sugar concentration of more than 4 g/L, which can be utilised by wine spoilage micro-organisms such as certain bacteria and other wild yeasts. Stuck and sluggish fermentations also increase the chances of oxidation due to the absence of the protective CO2 layer on the surface of the wine, which is formed during alcoholic fermentation. Another effect of stuck and sluggish fermentations is that valuable tank space is wasted due to the unexpected time consumption of these fermentation problems. Many factors during the winemaking process can be responsible for stuck and sluggish fermentations. In this thesis the different factors is discussed with the emphasis on the effect of the yeast strain. The way that certain yeast strains influence AAB and LAB numbers during fermentation and MLF through the production of inhibiting by-products such as medium chain fatty acids has not been investigated in detail in the past. Certain fungicides and pesticides that are used in vineyards to control pests (e.g. mildew) contain copper which can be inhibiting to yeast growth and alcoholic fermentation. Legal limits and withholding periods on these sprays are not always strictly obeyed and can lead to stuck and sluggish fermentations. This motivated us to evaluate the growth and fermentation activities of a selection of commercial wine yeasts in the presence of copper levels in the range of maximum legal limits. The effect of these commercial strains on the LAB and AAB numbers during alcoholic fermentation and MLF were also investigated. Our results showed that there was no significant difference on numbers of the AAB obtained from fermentations inoculated with different commercial wine yeast strains. However, with regards to the LAB numbers, one of the strains produced significantly more sulphur dioxide (SO2), which led to the inhibition of MLF in that wine. Our results further indicated which commercial yeast strains were capable of effectively fermenting high sugar musts and which strains were less effective. From the strains tested VIN13, N96 & L2056 were able to utilize fructose more effectively than NT50, RJ11 & D80. We could further distinguish between yeast strains that produced the lowest (VIN13 & RJ11) and the highest (WE372, NT50 & L2056) VA concentrations in must containing high sugar levels. Strains that were more tolerant against high copper levels were also identified. We tested six yeast strains in must with added copper (0.25 mM cu2+) in the form of CuSO4 .H2O. Three Cu2+-tolerant (D80, Collection Cepage Cabernet & NT50) yeast strains were distinguished from three less Cu2+-tolerant yeast strains (VIN13, NT112 & RJ11). This study made a valuable contribution in knowledge gained about commercially available wine yeast strains that can ferment effectively under certain stress conditions. Research such as this, where wine yeasts are evaluated to ferment more effectively during strenuous winemaking conditions, will be very beneficial to winemakers. / AFRIKAANSE OPSOMMING: Die produksie van gehalte wyn word deur verskillende faktore beïnvloed waarvan druifkwaliteit seker die belangrikste is. Die produksie van gehalte wyn is egter nie moontlik sonder goeie wynmaaktegnieke en effektiewe kwaliteitsbeheer nie. Kritieke kontrole punte (KKP) tydens die wynmaakproses moet dus geïdentifiseer word om sodoende ‘n verlaging in wynkwaliteit te vermy. Druiwemos het ‘n komplekse mikrobiologiese samestelling en bestaan uit verskillende mikroörganismes wat vooren nadelig vir wynkwaliteit kan wees, afhangende van die fisiese en chemiese toestande wat in die mos bestaan. Giste is verantwoordelik vir alkoholiese fermentasie, melksuurbakterieë (MSB) vir appelmelksuurgisting (AMG) en asynsuurbakterieë (ASB) vir die produksie van asynsuur vanaf etanol. Asynsuur word egter ook deur giste en MSB geproduseer en dra so by tot die vlugtige suurheid (VS) van ‘n wyn. Hierdie mikroörganismes kan mekaar op komplekse wyses beïnvloed deur o.a. te kompeteer vir voedingstowwe asook deur die produksie van inhiberende verbindings. Die meeste wynmakers maak gebruik van kommersiële gisrasse om alkoholiese fermentasies mee uit te voer. Gevalle van sogenaamde slepende en gestaakte alkoholiese fermentasies, waar suiker nie volledig na etanol en CO2 omgeskakel word nie, kom egter nog gereeld in die wynbedryf voor. In sulke gevalle neem die fermentasie gewoonlik langer as 21 dae om te voltooi met ‘n suiker konsentrasie van meer as 4 g/L wat in die wyn oorbly. Dit is nadelig vir wynkwaliteit aangesien dit nie net die kanse vir bederf deur bakterieë en giste verhoog nie, maar ook die kanse vir oksidasie verhoog a.g.v. die verlies van die beskermende CO2 lagie bo-oor die wyn. Hoe sekere gisrasse, ASB en MSB getalle gedurende fermentasie en AMG beïnvloed deur die produksie van inhiberende verbindings soos medium ketting vetsure en SO2, is ook nie baie in die verlede ondersoek nie. Sommige spuitstowwe wat gebruik word in die bekamping van swamsiektes bevat koper wat inhiberend kan wees vir gisgroei en alkoholiese fermentasie. Wetlike maksimum limiete en onthoudingsperiodes op spuitstofresidue word egter nie altyd gehoorsaam nie en kan lei tot slepende en gestaakte fermentasies. Dit het ons gemotiveer om ‘n seleksie van kommersiële gisrasse te evalueer in terme van gisgroei en fermentasie in die teenwoordigheid van kopervlakke naby die maksimum limiet. Ons resultate het gewys dat daar nie noemenswaardige verskille in AAB getalle tydens alkoholiese fermentasie tussen behandelings met verskillende kommersiële gisrasse was nie. Een van die gisrasse het wel noemenswaardig meer SO2 geproduseer wat gelei het tot inhibering van AMG in hierdie wyn. Ons het verder uitgewys watter kommersiële gisrasse instaat is om meer effektief in hoër suiker mos te fermenteer en watter van die rasse minder suksesvol was. Ons het ook rasse geïdentifiseer wat meer weerstandbiedend is teen hoë kopervlakke in mos en sodoende groter kans op ‘n suksesvolle fermentasie sal hê in mos wat koperresidue bevat wat afkomstig is van sekere spuitstowwe. Die effek van die ASB en MSB getalle gedurende fermentasie en AMG is ook ondersoek. Ons resultate het verder gewys watter kommersiële gisrasse instaat was om mos met hoë suikervlakke meer effektief te fermenteer. Vam die gisrasse wat getoets was het VIN13, N96 & L2056 fruktose meer effektief benut as NT50, RJ11 & D80. Ons kon verder onderskei tussen gisrasse wat die laagste (VIN13 & RJ11) en die hoogste (WE372, NT50 & L2056) vlakke van VS produseer in mos met hoë inisiële suikervlakke. Gisrasse wat meer tolerant was teen koperresidue in mos is ook geidentifiseer. Ons het ses gisrasse getoets in mos met bygevoegde koper (0.25 mM Cu2+) in die vorm van CuSO4 .5H2O. Daar is onderskei tussen drie Cu2+-tolerante (D80, Collection Cepage Cabernet & NT50) en drie minder Cu2+-tolerante gisrasse (VIN13, NT112 & RJ11). Hierdie studie lewer ‘n waardevolle bydrae in die invordering van kennis oor kommersieel beskikbare wyngisrasse wat meer effektief sal fermenteer onder sekere streskondisies wat in mos voorkom. Inligting soos hierdie is belangrik om die wynmaker se keuse uit die reeks bestaande kommersiële gisrasse te vergemaklik.

Thesis (MScAgric)--University of Stellenbosch, 2004. / ENGLISH ABSTRACT: Due to the extraordinary drought resistance of the grapevine, viticulture without irrigation in the winter rainfall coastal areas of South Africa is a feasible and commonly used practice. Wine quality is largely determined by the quality of the grapes from which it is made. Grapevine physiology is affected both directly and indirectly by water stress, which may vary according to soil type and prevailing atmospheric conditions. The water status of the grapevine can affect grape composition profoundly, either directly or indirectly, in either a positive or negative way, depending on the degree as well as the duration of water stress. There are three important factors involved in the development of water stress, namely the transpiration rate, the rate of water movement from the soil to the roots, and the relationship of soil water potential to leaf water potential. All three these factors are affected by atmospheric and/or soil conditions. In warm winelands such as South Africa (Western Cape), with a mediterranean climate which is characterised by a hot, dry summer period, the most important characteristic of soil is its ability to supply sufficient water to the grapevine during the entire growing season. Leaf water potential (Ψl) has gained wide acceptance as a fundamental measure of grapevine water status, and has been widely applied in viticultural research. Shortly before dawn, Ψl approaches equilibrium with soil water potential and reaches a maximum daily value. The study formed an integral part of a comprehensive, multi-disciplinary research project (ARC Infruitec-Nietvoorbij Project No. WW13/01) on the effect of soil and climate on wine quality, which commenced in 1993 and will be completed in 2004. This study was conducted during the 2002/03 growing season in two Sauvignon blanc vineyards situated at Helshoogte and Papegaaiberg, both in the Stellenbosch district, approximately nine kilometres apart. Two experiment plots, representing contrasting soil types in terms of soil water regime, were selected in each vineyard. At Helshoogte the two soils represented the Tukulu and Hutton forms, and the soils at Papegaaiberg were of the Avalon and Tukulu forms. The aim of this study was to determine the effect of atmospheric conditions and soil water status on the level of water stress in the grapevines for each soil at each locality, as well as the effect of grapevine water stress on yield and wine quality. This was done by determining and comparing the soil water status, soil water holding capacity of the soils and the evapotranspiration of the grapevines on the two different soils, at each of the two localities differing in mesoclimate and topography. The atmospheric conditions at the two localities during the 2002/03 season were also determined and compared to the long-term average atmospheric conditions, and the level of water stress of grapevines on each soil at each locality was measured. During the 2002/03 growing season, atmospheric conditions were relatively warm and dry in comparison to the long-term averages of previous seasons. These conditions accentuated the effects of certain soil properties that may not come forward during wetter, normal seasons. The usually wet Tukulu soil at Helshoogte was drier than expected during the 2002/03 season compared to the Hutton soil. Due to more vigorous growth on the Tukulu soil, grapevines extracted more soil water early in the season, leading to a low soil water matric potential and more water stress in the grapevines. Due to the higher vigour, resulting in more canopy shading, and more water stress, the dominant aroma in wines from the Tukulu soil was fresh vegetative. The Hutton soil maintained consistency with regards to both yield and wine quality compared to previous seasons. On the other hand the Tukulu soil supported a higher yield, but with lower than normal wine quality. The Avalon soil at Papegaaiberg maintained the highest soil water potential towards the end of the season, probably due to capillary supplementation from the sub-soil. Grapevines on the Tukulu soil at Papegaaiberg experienced much higher water stress than ones on the other three soils, especially during the later part of the season. This could be ascribed to a combination of factors, the most important being the severe soil compaction at a shallow depth, seriously limiting rooting depth and root distribution, which is detrimental to grapevine performance. Both the soil water status and atmospheric conditions played important roles in determining the amount of water stress that the grapevines experienced at different stages. The air temperature and vapour pressure deficit throughout the season were consistently lower at Helshoogte, the cooler terroir, compared to Papegaaiberg, the warmer terroir. At flowering, Ψl was lower for grapevines at Helshoogte than at Papegaaiberg, showing that diurnal grapevine water status was primarily controlled by soil water content. The difference in grapevine water status between the two terroirs gradually diminished until it was reversed during the post harvest period when Ψl in grapevines at Papegaaiberg tended to be lower compared to those at Helshoogte. The relatively low pre-dawn Ψl at Helshoogte indicated that the grapevines were subjected to excessive water stress resulting from the low soil water content. However, grapevines at Helshoogte did not suffer material water stress (i.e. Ψl < -1.20 MPa) during the warmest part of the day, suggesting that partial stomatal closure prevented the development of excessive water stress in the grapevines. This suggests that low pre-dawn Ψl values do not necessarily imply that grapevines will experience more water stress over the warmer part of the day, or visa versa. This does not rule out the possibility that side-effects of partial stomatal closure, such as reduced photosynthesis, can have negative effects on grapevine functioning in general. These results also suggest that measurement of diurnal Ψl cycles at various phenological stages is required to understand and quantify terroir effects on grapevine water status. / AFRIKAANSE OPSOMMING: Danksy die droogteweerstand van die wingerdstok is die verbouing van wingerde sonder besproeiing ‘n praktiese en algemene verskynsel in die winterreënval-areas van Suid-Afrika. Wynkwaliteit word grootliks bepaal deur die kwaliteit van die druiwe waarvan dit gemaak word. Wingerdfisiologie word direk en indirek beïnvloed deur waterstres, wat kan varieer volgens die grondtipe en die heersende atmosferiese toestande. Die waterstatus van die wingerdstok beïnvloed druifsamestelling, direk of indirek, en positief of negatief, afhangend van die graad en tydsduur van die waterstres. Daar is drie belangrike faktore betrokke by die ontwikkeling van waterstres, naamlik die transpirasietempo, die tempo van waterbeweging vanaf die grond na die wortels, en die verhouding tussen die grondwatermatrikspotensiaal tot blaarwaterpotensiaal. Al drie die faktore word beïnvloed deur die atmosferiese en/of grondtoestande. In warm wynboulande soos Suid-Afrika (Weskaap), met ‘n meditereense klimaat wat gekarakteriseer word deur ‘n warm, droë somerperiode, is die belangrikste eienskap van grond die vermoë om voldoende water aan die wingerdstok te verskaf gedurende die hele seisoen. Blaarwaterpotensiaal (Ψl) het wye aanvaarding bekom as die fundamentele meting van wingerstokwaterstatus, en word wyd toegepas in wingerdkundige navorsing. Kort voor sonsopkoms, nader Ψl ‘n ewewig met die grondwatermatrikspotensiaal en bereik ‘n maksimum daaglikse waarde. Die studie vorm ‘n integrale deel van ‘n omvattende, multi-dissiplinêre navorsingsprojek (ARC Infruitec-Nietvoorbij Projek No. WW13/01) op die effek van grond en klimaat op wynkwaliteit, wat in 1993 in aanvang geneem het en in 2004 afgehandel sal word. Hierdie studie is uitgevoer gedurende die 2002/03 seisoen in twee Sauvignon blanc wingerde geleë by Helshoogte en Papegaaiberg, beide in die Stellenbosch distrik, ongeveer nege kilometer van mekaar. Twee eksperimentele persele, elkeen verteenwoordigend van kontrasterende grondtipes in terme van grondwaterregime, is geselekteer in elke wingerd. By Helshoogte word die twee gronde verteenwoordig deur die Tukulu en Hutton grondvorms, en die gronde by Papegaaiberg is van die Avalon en Tukulu vorms. Die doel van die studie was om die effek van atmosferiese toestande en grondwaterstatus op die wingerdstok se waterstatus vir elke grond by die twee lokaliteite te bepaal, sowel as die effek van die wingerdstok se waterstatus op die opbrengs en wynkwaliteit. Dit is gedoen deur die grondwaterstatus, die grondwaterhouvermoë, sowel as die evapotranspirasie van die wingerdstokke op die twee verskillende gronde by elk van die twee lokaliteite, wat verskil in mesoklimaat en topografie, te bepaal en vergelyk. Die atmosferiese toestande by die twee lokaliteite gedurende die 2002/03 seisoen is ook bepaal en vergelyk met die langtermyn gemiddelde atmosferiese toestande. Die vlakke van waterstres in wingerdstokke op elke grond by elke lokaliteit is ook gemeet. Gedurende die 2002/03 groeiseisoen, was die atmosferiese toestande relatief warm en droog in vergelyking met die langtermyn gemiddeldes van vorige seisoene. Hierdie kondisies aksentueer die effek van sekere grondeienskappe wat nie noodwendig na vore kom gedurende normale, natter seisoene nie. Die gewoonlike nat Tukulu grond by Helshoogte was droër as verwag gedurende 2002/03 in vergelyking met die Hutton grond. As gevolg van sterker groekrag op die Tukulu grond, het wingerdstokke meer grondwater onttrek vroeg in die seisoen, wat gelei het tot ‘n lae grondwatermatrikspotensiaal en meer waterstres in die wingerdstokke. Die sterker groeikrag het meer beskaduwing van die lower asook meer waterstres veroorsaak, wat gelei het daartoe dat die dominante aroma in wyne vanaf druiwe op die Tukulu grond vars vegetatief was. Die Hutton grond het bestendig gebly in terme van opbrengs en wynkwaliteit in vergelyking met vorige seisoene. Daarteenoor het die Tukulu grond ‘n hoër opbrengs gelewer, maar met laer as gewoonlike wynkwaliteit. Die Avalon grond by Papegaaiberg het die hoogste grondwatermatrikspotensiaal behou tot die einde van die seisoen, heelwaarskynlik a.g.v. kapillêre aanvulling vanuit die ondergrond. Wingerdstokke op die Tukulu grond by Papegaaiberg het heelwat meer waterstres ondervind as op die ander drie gronde, veral later in die seisoen. Dit kan toegeskryf word aan ‘n kombinasie van faktore, die belangrikse daarvan die erge grondkompaksie vlak in die grond, wat worteldiepte en -verspreiding ernstig beperk het, wat op sy beurt nadelig is vir wingerdprestasie. Beide die grondwaterstatus en atmosferiese toestande het ‘n belangrike rol gespeel in die bepaling van die hoeveelheid waterstres wat die wingerdstok op verskillende stadiums ondervind het. Die lugtemperatuur en waterdampdruktekort was regdeur die seisoen laer by Helshoogte, die koeler terroir, as by Papegaaiberg, die warmer terroir. Gedurende blom was die Ψl laer vir wingerdstokke by Helshoogte as by Papegaaiberg, wat daarop wys dat daaglikse wingerdstok waterstatus hoofsaaklik deur die grondwaterinhoud bepaal was. Die verskil in wingerdstok waterstatus tussen die twee terroirs het geleidelik verminder totdat dit omgekeer was gedurende die na-oes periode toe Ψl in wingerdstokke by Papegaaiberg geneig het om laer te wees in vergelyking met die by Helshoogte. Die relatiewe lae voorsonop Ψl by Helshoogte het daarop gedui dat die wingerdstokke aan oormatige waterstres onderwerp was. Die wingerdstokke by Helshoogte het egter nie materiële waterstres (i.e. Ψl < -1.20 MPa) gedurende die warmste gedeelte van die dag ondervind nie, wat aandui dat gedeeltelike huidmondjiesluiting plaasgevind het om die ontwikkeling van oormatige waterstres te verhoed.Dit dui aan dat lae voorsonop Ψl waardes nie noodwendig impliseer dat wingerdstokke meer waterstres gedurende die warmste gedeelte van die dag sal ondervind nie, of visa versa. Dit sluit nie die moontlikheid uit dat negatiewe neweeffekte van gedeeltelike huidmondjiesluiting, soos ‘n vermindering in fotosintese, ‘n negatiewe effek kan hê op die wingerdstok se funksionering in die algemeen nie. Hierdie resultate stel voor dat die meting van daaglikse Ψl siklusse gedurende verskeie fenologiese stadia benodig word om die effek van terroir op die wingerdstok se waterstatus te verstaan en te kwantifiseer.

Thesis (MScAgric)--University of Stellenbosch, 2004. / ENGLISH ABSTRACT: The distinctive varietal flavour of wines is a combination of absolute and relative concentrations of chemical compounds. Volatile compounds are responsible for the odour of wine and non-volatiles cause the sensation of flavour. Accompanying these senses, a third, tactile, sense of ‘mouth-feel’ is recognizable. This forms the complete organoleptic quality of wine. Several hundred different compounds are simultaneously responsible for the odour release in wine, and since there is no real character impact compound, the aroma of wine can be described as a delicate balance of all these compounds. One of the most important groups of volatiles is the monoterpenes, which play a role in both aroma and flavour. This is especially significant for the Muscat varieties, but these flavour compounds are also present in other non-muscat grape varieties, where they supplement the varietal aroma. Monoterpenes occur in wine as free, volatile and odorous molecules, as well as flavourless non-volatile glycosidic complexes. The latter slowly releases monoterpenes by acidic hydrolysis, but the impact on varietal aroma is considered insufficient for wines that are consumed young. It is therefore important to supplement the release mechanism, in order to enhance the varietal aroma of the wine. The enzymatic hydrolysis mechanism functions in two successive steps: firstly, depending on the precursor, the glycosidic linkage is cleaved by α-L-arabinofuranosidase, α-L-rhamnosidase, β-D-xylosidase or β-D-apiosidase. The second step involves the liberation of the monoterpene alcohol by a β-glucosidase. This enzymatic hydrolysis does not influence the intrinsic aromatic characteristics of the wine, as opposed to acid hydrolysis. Pectolytic enzymes play an important role in cell elongation, softening of tissue and decomposition of plant material. These enzymes are used to improve juice yields, release colour and flavour compounds from grape skins, as well as improve clarification and filterability. Pectolytic enzymes work synergistically to break down pectins in wine. Protopectinase produce water-soluble and highly polymerised pectin substances from protopectin, it acts on non-methylated galacturonic acid units. Pectin methylesterase split methyl ester groups from the polygalacturonic chain. Polygalacturonase break down the glycosidic links between galacturonic acid units. Pectin and pectate lyases have a β-eliminative attack on the chain and it results in the formation of a double bond between C4 and C5 in the terminal residues. From the above it can be seen that enzymes play a pivotal role in the winemaking process. Unfortunately, in winemaking a lot of factors can influence the effects of enzymes. One possible factor in the wine medium is the presence of acidprotease, from yeast and/or fungal origin. This type of enzyme utilizes other enzymes as substrates and renders them useless. Pure enzyme preparations were used to study the interactions of a yeast acid-protease and a report activity (β-glucosidase) in vitro. A bottled wine and a buffer were used as in vitro conditions. Enzyme assays were performed to determine the relative activity over a number of days. The results indicated that even though both enzymes showed activity in both the media, the yeast protease did not have any significantly affect on the report activity. Subsequently wine was made from Sauvignon blanc grapes, with varying enzyme preparation additions. Enzyme assays were performed during the fermentation; and chemical, as well as sensory analysis were done on the stabilized wine. The results confirmed that the yeast protease did not have any significant affect on the report activity in these conditions. The protease’s inability to affect the report activity seems unlikely due to the fact that it is active at a low pH range and has been suggested as the only protease to survive the fermentation process. It seems possible that a winerelated factor, possibly ethanol, is responsible. Thus it seems that yeast protease does not threaten the use of commercial enzymes in the winemaking process in any significant way. Future work would entail more detailed enzyme studies of interactions between protease, both from yeast and fungal origin, and other report activities in specified conditions. The degradation capability could be directed towards unwanted enzyme activities that cause oxidation and browning of the must. The characterization of interactions between protease and β-glucosidase activities may hold key to producing wines with enhanced aroma and colour potential, as well as the elimination of unwanted enzyme activities. / AFRIKAANSE OPSOMMING: Die herkenbare kultivar karakter van wyn is ‘n kombinasie van absolute en relatiewe konsentrasies van verskeie chemiese komponente. Vlugtige komponente is verantwoordelik vir die geur, of aroma, van wyn en die nie-vlugtige komponente veroorsaak die sensasie van smaak. ‘n Derde, fisiese sensasie, die ‘mondgevoel’, is ook herkenbaar. Dit vorm die omvattende organoleptiese kwaliteit van die wyn. ‘n Paar honderd verskillende komponente is gelyktydig verantwoordelik vir die aroma vrystelling in wyn en omdat daar geen werklike karakter ‘impak’ komponent is nie, kan die aroma van wyn beskryf word as ‘n delikate balans van al die betrokke komponente. Een van die mees belangrike groepe vlugtige komponente is die monoterpene wat ‘n rol speel in beide aroma en smaak. Dit is veral belangrik by Muskaat kultivars, maar hierdie aroma komponente is ook teenwoordig in niemuskaat druif kultivars, waar hulle bydra tot die kultivar karakter en aroma. Monoterpene kom in wyn voor as vry, vlugtige en aromatiese molekules en in geurlose, nie-vlugtige glikosidies-gebonde komplekse. Die gebonde vorm word stadig vrygestel deur ‘n suurhidrolise, maar dit word as onvoldoende beskou vir wyne wat vroeg gedrink word. Dit is dus belangrik dat die vrystelling van geurstowwe verhoog word om die kultivar karakter van die wyn te versterk. Die ensiematiese hidrolise proses behels twee opeenvolgende stappe: eerstens, afhangende van die aard van die voorloper, word die glikosidiese verbinding deur α-L-arabinofuranosidase, α-Lramnosidase, β-D-xilosidase, of β-D-apiosidase gebreek. In die tweede stap word die monoterpeen-alkohol deur β-glukosidase vrygestel. Hierdie ensiematiese afbraak proses verander nie die intrinsieke aromatiese kenmerke van die wyn, soos met suurhidrolise die geval is nie. Pektolitiese ensieme speel ‘n fundamentele rol in selverlenging, sagwording en afbraak van plant materiaal. Hierdie ensieme word gebruik om sap opbrengs te verhoog, aroma en smaak komponente vry te stel uit die doppe, asook om sapverheldering en filtrasie te verbeter. Die pektolitiese ensieme werk op ‘n sinergistiese wyse om pektien in wyn af te breek. Protopektinase produseer wateroplosbare en hoogs gepolimeriseerde pektien uit protopektien, slegs uit niegemetileerde galakturoonsuur eenhede. Pektien metielesterase verwyder metielester groepe van die poligalakturoonsuurketting. Die glikosidiese bindings tussen galakturoonsuur eenhede word deur poligalakturonase afgebreek. Pektien- en pektaat-liase het ‘n β-eliminasie aanslag op die ketting en as gevolg daarvan word dubbelbindings tussen C4 en C5 in die terminale residue gevorm. Vanuit bogenoemde is dit dus duidelik dat ensieme ‘n kardinale rol speel in die wynbereidingsproses. Ongelukkig is daar ‘n verskeidenhied van faktore wat die werking van ensieme in die wynbereidingsproses kan beïnvloed. Een moontlike faktor is die teenwoordigheid van ‘n suur-protease, van fungisidiese en/of gis oorsprong, in die wynmedium, omdat dit ander ensieme as substraat kan benut en degradeer. Suiwer ensiem preparate is gebruik om die ensiem interaksie tussen ‘n gis suur-protease en ‘n verslag aktiwiteit (β-glukosidase) in vitro te ondersoek. ‘n Gebotteleerde wyn en ‘n buffer is gebruik om die in vitro kondisies na te boots. Relatiewe ensiem aktiwiteit is ontleed oor ‘n aantal dae. Beide die ensieme het aktiwiteit getoon in die media, maar gis protease het geen statisties beduidende invloed gehad op die aktiwiteit van die verslag ensiem nie. Daaropvolgend is wyn berei van Sauvignon blanc druiwe, met verskillende ensiempreparaat toevoegings. Die ensiemaktiwiteit is deurlopend tydens fermentasie gemeet. Na afloop van stabilisasie is chemiese, sowel as sensoriese ontledings op die wyn gedoen. Die resultate het bevestig dat gis protease, onder hierdie kondisies, geen beduidende invloed op die verslag aktiwiteit gehad het nie. Die protease se onvermoë om die verslag aktiwiteit beduidend te beinvloed blyk onwaarskynlik aangesien die suurprotease aktief is by lae pH vlakke en dit as die enigste protease voorgestel is wat die fermentasie proses kan oorleef. Dit blyk asof ‘n wyn-verwante faktor, moontlik etanol, hiervoor verantwoordelik kan wees. Dus hou protease geen gevaar in vir die gebruik van kommersiële ensieme in wynbereiding nie. Navorsing kan in die toekoms fokus op meer gedetailleerde ensiem interaksie studies tussen protease en ander ensiem aktiwiteite, in gespesifiseerde kondisies. Die degradasie kapasiteit kan moontlik aangewend word om ongewenste ensiem aktiwiteite, wat byvoorbeeld oksidasie en verbruining veroorsaak, te verminder. Die karakterisering van die interaksies tussen protease en β-glukosidase kan dus die sleutel wees tot die produksie van wyne met verhoogde aroma potensiaal, asook die eliminasie van ongewenste ensiematiese aktiwiteite.

Thesis (MSc)--University of Stellenbosch, 2005. / ENGLISH ABSTRACT: In the field of agriculture, plant pathogens are a major concern because of the severe damage these organisms cause to crops yearly. Fundamental studies regarding plant pathogens and their modes of action made it possible for researchers in the field of molecular biology to investigate pathogens further on a molecular level. Botrytis cinerea, has been used to great effect as a model system to investigate various aspects regarding pathogenesis, also on a molecular level. Molecular research done on B. cinerea over the last few years has shown that the endopolygalacturonases (EPGs) of this fungus are key role players in pathogenesis. This hydrolytic enzyme family of six members, encoded by the Bcpg1-6 genes, are important in breaking down the complex cell wall polymers of host plants, enabling the fungus to penetrate its host sufficiently. It has been shown that both BcPG1 and 2 are crucial for virulence of B. cinerea. A leucine-rich repeat inhibitor protein situated in the cell wall of various plant species, the polygalacturonase-inhibiting protein (PGIP), has been proven to interact with and inhibit EPGs, and thus the necrotic actions of B. cinerea. From literature it was clear that specific data regarding individual interactions of fungal EPGs with PGIPs are lacking currently. Furthermore, most experiments regarding the effects of EPG as well as interaction and inhibition studies of EPGs and PGIPs, rely on in vitro methods, without the possibility to contextualize the results on an in vivo or in planta level. The scope of this study was to specifically address the issues of individual EPG:PGIP interactions and the use of possible in vivo methodology by using EPGs from a highly virulent South African strain of B. cinerea and the grapevine VvPGIP1 that has been previously isolated in our laboratory. This PGIP, originally isolated from Vitis vinifera cv Pinotage, has been shown to inhibit a crude EPG extract from this strain with great efficiency. The approach taken relied on heterologous over-expression of the individual Bcpg genes and the isolation of pure and active enzymes to evaluate the inhibition of the EPGs with VvPGIP1. The genes were all successfully over-expressed in Saccharomyces cerevisiae with a strong and inducible promoter, but active enzyme preparations have been obtained only for the encoding Bcpg2 gene, as measured with an agarose diffusion assay. The in vitro PGIP inhibition assay is also based on the agarose diffusion assay and relies on activity of the EPGs to visualize the inhibiting effect of the PGIP being tested. The active EPG2, however, was not inhibited by VvPGIP1 when tested with this assay. The EPG encoding genes from B. cinerea were transiently over-expressed also in Nicotiana benthamiana by using the Agrobacterium-infiltration technique. Transgene expression was confirmed by Northern blot analysis and EPG-related symptoms were observed five to eight days post-infiltration. Differential symptoms appeared with the various EPGs, providing some evidence that the symptoms were not random events due to the infiltration or a hypersensitive response. Moreover, the symptoms observed for EPG2 was similar to those that were reported recently by another group on the same host. In spite of the expression data and the clear symptoms that developed, active preparations, as measured with the agarose diffusion plate asay, could only be obtained for EPG2 again. In our search for a possible in vivo method to detect and quantify EPG activity and inhibition by PGIPs, we tested and evaluated a technique based on chlorophyll fluorescence to detect the effect of EPGs on the rate of photosynthesis. Our results showed that the over-expression of these genes reduced the rate of electrons flowing through photosystem II, indicating metabolic stress occurring in the plant. We used the same technique to evaluate possible interaction between VvPGIP1 respectively with BcPG1 and 2 and found that the co-expressing of the Vvpgip1 gene caused protection of the infiltrated tissue, indicating inhibition of EPG1 and 2 by VvPGIP1. For EPG2, the observed interaction and possible inhibition by VvPGIP1 is the first report to our knowledge of an interaction between this specific EPG2 and a PGIP. Moreover, to further elucidate the in planta interaction between VvPGIP1 and the EPGs from the South African B. cinerea strain, we tested for possible interactions by making use of a plant two-hybrid fusion assay, but the results are inconclusive at this stage. Previous studies in our laboratory have shown that several natural mutations exist between PGIP encoding genes from different V. vinifera cultivars. Based on this finding and the fact that these natural mutations could result in changes with regard to EPG inhibition and ultimately disease susceptibility, we isolated an additional 37 PGIP encoding genes from various grapevine genotypes, some of which are known for their resistance to pathogens. Combined, these results make a valuable contribution to understand plant pathogen interactions, specifically in this case by modeling the interactions of pathogen and plant derived proteins. The possibility to use in vivo methods such as chlorophyll fluorescence to follow these interactions on an in planta level, provides exciting possibilities to strenghten and contextualize in vitro results. / AFRIKAANSE OPSOMMING: Plantpatogene organismes veroorsaak jaarliks erge skade aan landbougewasse en word dus as ’n ernstige probleem in die landbousektor beskou. Diepgaande studies wat handel oor plantpatogene en hul metodes van infeksie het dit vir molekulêre bioloë moontlik gemaak om patogene nou ook op molekulêre vlak verder te bestudeer. Botrytis cinerea is baie effektief as modelsisteem gebruik om verskeie aspekte van patogenese verder te bestudeer, ook op ‘n molekulêre vlak. Molekulêre navorsing op B. cinerea, het getoon dat die endopoligalakturonases (EPGs) van dié swam kernrolbelangrik in patogenese is. Hierdie sesledige hidrolitiese ensiemfamilie word gekodeer deur die Bcpg1-6 gene en is belangrik vir die afbraak van die komplekse selwandpolimere van plantgashere, om suksesvolle gasheerpenetrasie te veroorsaak. Daar is aangetoon dat beide BcPG1 en 2 essensieël vir virulensie van die patogeen is. ’n Leusienryke-herhalings inhibitorproteïen wat in die selwand van verskeie plantspesies voorkom, die poligalakturonase-inhiberende proteïen (PGIP), het interaksie met en inhibeer EPGs en gevolglik ook die nekrotiserende aksies van B. cinerea. Uit die literatuur is dit duidelik dat spesifieke inligting aangaande individuele interaksies van fungiese EPGs met PGIPs tans nog ontbreek. Verder word daar op in vitro metodologie staatgemaak wannneer die effekte van EPGs asook die interaksie en inhibisie met PGIPs bestudeer word, sonder om die konteks van die in vivo- of in planta-omgewing in ag te neem. Die fokus van hierdie studie was om aspekte van individuele EPG:PGIP interaksies, asook die moontlike gebruik van in vivo metodologie te bestudeer deur EPGs, afkomstig van ’n hoogs virulente Suid-Afrikaanse ras van B. cinerea en die wingerd VvPGIP1, wat vroeër in ons laboratorium geïsoleer is, te gebrruik. Hierdie PGIP wat uit Vitis vinifera cv Pinotage geïsoleer is, inhibeer ’n kru EPG-ekstrak van bogenoemde ras baie effektief. Die benadering wat gevolg is het op die ooruitdrukking van die individuele Bcpg-gene in heteroloë sisteme staatgemaak en die gevolglike isolering van suiwer en aktiewe ensieme om EPG-inhibisie deur VvPGIP1 te beoordeel. Al die gene is suksesvol in Saccharomyces cerevisiae ooruitgedruk onder ’n sterk induseerbare promotor, maar volgens ’n agarose-diffundeerbare toets kon aktiewe ensiempreparate slegs vir die enkoderende Bcpg2 verkry word. Die in vitro PGIP-inhibisie toets is ook op die gemelde toets gebasseer en vereis EPG-aktiwiteit om die inhiberende effek van die PGIP, te visualiseer. Die aktiewe EPG2 is egter nie deur VvPGIP1 geïnhibeer met die aanleg van hierdie toets nie. Die EPG-enkoderende gene van B. cinerea is ook tydelik in Nicotiana benthamiana ooruitgedruk deur gebruik te maak van ’n Agrobacterium-infiltrasietegniek. Transgeenuitdrukking kon met die Noordelike kladtegniek bevestig word en EPG-verwante simptome is vyf tot agt dae na infiltrasie waargeneem. Verskillende simptome vir die verskillende EPGs is waargeneem, wat aanduidend is dat die simptome nie lukrake gevolge van die infiltrasies, of ’n hipersensitiewe respons is nie. Verder kon die simptome wat EPG2 vertoon het, gekorreleer word met dié wat onlangs deur ’n ander groep op dieselfde gasheer waargeneem is. Ten spyte van die ekspressiedata en die waargenome simptome, kon aktiewe ensiempreparate op die agarose-diffundeerbare toets, weereens slegs vir EPG2 waargeneem word. ’n Metode wat gebasseer is op chlorofilfluoressensie is getoets en geëvalueer as ’n moontlike in vivo metode om EPG aktiwiteit en inhibisie deur PGIPs waar te neem en te kwantifiseer. Die resultate het bevestig dat die ooruitdrukking van hierdie gene die elektronvloeitempo deur fotosisteem II verminder het wat ’n aanduiding is dat metaboliese stres in die plant heers. Dieselfde tegniek is gebruik om die moontlike interaksies tussen BcPG1 en 2 en VvPGIP1 te bestudeer en het aangetoon dat die mede-uitdrukking van die Vvpgip1-geen aanleiding gee tot ’n beskermende effek van die geinfiltreerde weefsel, wat aanduidend is van inhibisie van EPG1 en 2 deur VvPGIP1. In die geval van EPG2 is hierdie interaksie en moontlike inhibisie met ’n PGIP die eerste waarneming in die verband. In ’n verdere poging om die in planta-interaksie tussen VvPGIP1 en die EPGs van die Suid-Afrikaanse B. cinerea ras uit te klaar, is ’n plantgebasseerde twee-hibriede toets aangelê, maar geen klinkklare resultate kon verkry word nie. Vorige werk het bevestig dat verskeie natuurlike mutasies in PGIP-enkoderende gene, afkomstig van verskillende V. vinifera kultivars, voorkom. Hierdie resultaat en die feit dat hierdie mutasies verskille in EPG inhibisie en uiteindelik vatbaarheid vir siektes kan beïnvloed, het aanleiding gegee tot die isolering van ’n verdere 37 PGIP-enkoderende gene uit ‘n verskeidenheid druifplantgenotipes, sommige waarvan juis bekend vir hul weerstand teen patogene is. Die gekombineerde resultate wat in dié studie verkry is, maak ’n waardevolle bydrae tot die verstaan van plant-patogeeninteraksies, spesifiek met die modelering van interaksies van patogeen- en plantgebasseerde proteïene. Die moontlikheid om in vivo-metodes soos chlorofilfluoressensie te gebruik in in planta-analises, is besonder bemoedigend om in vitro-resultate te versterk en ook in konteks te plaas.

Thesis (MscAgric (Viticulture and Oenology))--University of Stellenbosch, 2007. / A common phenomenon in most South African vineyards, especially in the Western Cape region, is that of within vineyard variation. This variation phenomenon is caused by an array of controllable and non-controllable factors that interact with each other to affect vine vigour. Controllable factors can be managed by the grape grower, while the non-controllable factors have to be managed in the planning process in order not to negatively affect productivity or product quality. The main goal of any grape grower is to optimise vine performance in an attempt to achieve the best possible yield while at the same time allowing vines to optimally ripen grapes towards optimal wine quality. A grape grower has to use every possible means and technique available to him in order to manage his vineyards in such a manner as to achieve this goal. In the past, it was difficult to visualize the extent and distribution of vigour variation in vineyards, but with modern technological improvements in the field of remote sensing, grape growers are able to identify and specify different vigour levels within a vineyard. When remote sensing is applied in a vineyard, the grape grower can identify certain areas that may need more specific attention than others. Consequently, managerial decisions based on detailed information can be made in an attempt to improve the general condition and performance of a vine. Not only can the acquired information be used to plan managerial actions throughout the season, but it can also be used to plan and devise harvest strategies. Some areas in a vineyard may be at a certain point in the ripening process and need to be harvested, while grapes from other areas still need to develop the wanted flavours. One managerial action applied at véraison by some grape growers, is that of crop thinning. Different vigour areas can now be subjected to various crop thinning actions in an attempt to determine the best crop load for a vigour level. With this in mind, two studies were launched to firstly investigate the interaction between vine vigour and harvest dates; and secondly to investigate the interaction between vine vigour and crop load and how their combined interaction might influence a vine’s characteristics, grape composition and wine quality. Vigour variation was firstly identified through multispectral aerial imagery, and then visually verified by visits to the experimental vineyards. The multispectral aerial image was then “orthorectified” in order to produce a classified multispectral image. The image was classified through different colour codes that were assigned to the different vigour levels to clearly distinguish between them. A series of vegetative and reproductive measurements were conducted to try and establish if any correlations could be obtained of the interaction between vine vigour, different harvest dates and crop loads. In order to verify differences in vine vigour, underlying causes were also determined through soil analyses of which chemical analysis, bulk density, porosity, as well as root penetration and distribution were determined. Vegetative measurements that were conducted for both studies indicated good correlations between the different vigour levels and the image classifications. The results also identified the effect that topping (mechanical or manual) had on the main and lateral leaf areas. Reproductive measurements throughout the season, in the form of berry sampling, showed changes in berry composition and accentuated the effects of the different treatments, which could also be confirmed through sensorial analysis of the wines. The results also emphasized the need to not only make use of one of two chemical parameters to identify grape ripeness, but to incorporate a number of parameters, such as sugar, pH and acid levels. From the varying grape chemical characteristics, a wine style can be identified that might carry the approval of the winemaker for the production of a specific type of wine. Soil studies of both vineyards also gave important evidence for the causes of vigour variation. The data collected will hopefully provide grape growers with information that will enable them to make educated decisions concerning grape production and how vigour, in conjunction with different harvest dates and crop loads, will enable them to produce fruit of good quality and, so doing, improve their financial position.

Thesis (MscAgric (Viticulture and Oenology))--Stellenbosch University, 2008. / The effect of various irrigation strategies on grapevine water relations during the berry ripening period was investigated in a Shiraz/Richter 99 vineyard. Comparisons between different irrigation strategies (full/seasonal, véraison+post véraison, post véraison and no irrigation) were made. During the day, the seasonally irrigated vines experienced less water stress than the deficit treatments. Non-irrigated vines seemed to maintain higher diurnal leaf water potentials. Lower leaf water potentials indicated lower water contents in the vegetative and reproductive tissue. Full irrigation seemed to stimulate primary shoot length. Longer water deficit induced earlier and more complete shoot maturation (reserve accumulation). Re-distribution of leaf area on the shoot may occur when vines are subjected to water deficit. Extended water deficit seemed to induce earlier and restricted water loss from vegetative tissue. The water relations were reflected in the berry size. Irrigation during ripening seemed to induce a continuation of berry water loss. Transpiration losses were apparently much higher in fully irrigated vines whereas stomatal control efficiently maintained water relations in non-irrigated vines. Water deficit seemed to have enhanced the soluble solid accumulation. Irrigation treatments did not seem to affect the titratable acid and pH. The post véraison irrigation in particular seemed to favour a wide window for harvesting. Irrigation at post véraison and especially véraison+post veraison seemed to have a greater effect on the synthesis and extraction of phenolics, anthocyanins and tannins in the berry skins. Different irrigation strategies may affect grapes in such a way that different wine styles are obtained.

Thesis (MscAgric (Viticulture and Oenology))--Stellenbosch University, 2008. / Over the years, different techniques have been used to legally reduce the ethanol content of wines. Several physical processes are available for producing wines with less alcohol. Despite their efficacy, these treatments have a capital and operational cost influence. They can also affect the concentration of other wine components. On the other hand, vast amount of research has been conducted through genetic modification of wine yeast strains in order to reduce the ethanol yield of Saccharomyces cerevisiae by diverting sugar metabolism towards various byproducts. However, genetically modified yeasts are not currently accepted in most wine industries worldwide, including South Africa. Therefore, other approaches need to be envisaged. Commercial enzymes are commonly added during winemaking. Most enzymes essential for vinification naturally occur in grapes, but are inefficient under pH and sulphur levels associated with winemaking. Enzymes of fungal origin are resistant to such conditions. The most widely used commercial enzymes include pectinases, hemicellulases, glucanases and glycosidases. With the exception of glucanases, produced by Trichoderma harzianium, all the other enzymes are produced by Aspergillus niger. In this study, the possibility of using Gluzyme Mono® 10.000 BG (Gluzyme) (Novozymes, South Africa) to reduce the glucose content of synthetic grape must and grape must before fermentation in order to produce wine with a reduced alcohol content was investigated. Gluzyme is a glucose oxidase preparation from Aspergillus oryzae, currently being used in the baking industry. Glucose oxidase catalyses the oxidation of glucose to gluconic acid and hydrogen peroxide in the presence of molecular oxygen. Gluzyme was initially used in synthetic grape must where different enzyme concentrations and factors influencing its activity were investigated for its use in winemaking. The results showed that up to 0.5% v/v less alcohol were obtained using an enzyme concentration of 20 kU compared to the control. This reduction in alcohol was increased to 1 and 1.3% v/v alcohol at pH 3.5 and pH 5.5 respectively in aerated synthetic grape must using 30 kU enzyme. Secondly, Gluzyme trials were carried out using Pinotage grape must. Gluzyme treated wines after fermentation contained 0.68% v/v less alcohol than the control samples at 30 kU enzyme. Colour and volatile flavour compounds of treated wine did not differ significantly from the untreated samples. Lower free anthocyanin and total phenol concentrations in treated than control samples were observed, possibly due to the hydrogen peroxide oxidation which could have led to polymerisation. The present study has clearly demonstrated that Gluzyme may be used in winemaking to produce reduced-alcohol wine without affecting its colour and aroma compounds. The enzyme in its current form is however, not ideal for winemaking; other forms such as liquid or powder form should be considered if the enzyme is to be used under winemaking conditions. Future work should focus on evaluating the potential new form of the enzyme and studying the effects of Gluzyme in various grape must in semi-industrial scale. A tasting panel should also evaluate its impact on the organoleptic properties and the overall quality of the resulting wines.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: Grapevine water status is considered to be the most important factor limiting plant growth and production in the Mediterranean zones. In these regions with limited summer rainfall and limited water resources for irrigation grapevines may experience water deficits for an extended period of time. The demand of water for agriculture is constantly increasing, and will continue to do so due to the rise in the world population and to the effects of climate change on rainfall and evaporative demand in these regions. The Western Cape wine region is also classified as Mediterranean and grapevines grown in this region are often exposed to water “stress” conditions due to high evaporative demand and low water availability in the soil. Plant water status of grapevines may dependent on, amongst other factors, the water potential of soil layers close to the root system, canopy size and evaporative demand. The canopy size of a grapevine can inherently be seen as a measure of grapevine vigour, and vigour variation among grapevines within a vineyard is a common phenomenon in the Western Cape. The importance of the contributions from several factors causing vigour variation within vineyards is still a subject of debate. This may be largely ascribed to the significant amount of variability in vineyards that researchers have to deal with during viticultural studies. However, the recent advances in remote sensing technology have established new methods to assess grapevine vigour variability. In the face of the recognized variation within vineyards and the importance of a sustained grapevine water status, for wine grape productivity and -quality, it is alarming to think that a vineyard block is generally managed as a homogeneous entity when it comes to irrigation scheduling. What is more alarming is the assumption that grape, juice and wine quality will be homogeneous throughout a vineyard block – even without irrigation. With this in mind, a study was conducted to study the interaction between grapevine vigour and grapevine water status within a commercial vineyard with variable vigour by implementing various irrigation regimes. Vigour variation was identified through multispectral aerial imagery and plant-based water status determinants were used to assess grapevine water status in plots of differing vigour within the vineyard. Soil water status was also assessed, and vegetative growth quantified to ultimately determine the variability in vigour and its possible contribution to the variability through the water status of the plant. Reproductive growth was monitored continually before evaluating the effect of water status and grapevine vigour on grape composition and subsequent wine quality. The various methods used to evaluate grapevine vigour showed good correspondence. Pruning mass measured at the end of the season confirmed leaf area measurement (main leaves and lateral leaves) during vegetative growth, and corresponded well, in terms of main vigour classifications with the NDVI images collected. Berry weight and volume responded to the various classifications, with a decrease in water deficits from one classification to the next accompanying an increase in berry weight and volume. Analyses of the berry composition and wines showed statistically significant differences between the classifications. This was found for sugar content per berry, total phenols, total red pigment, malic acid, nitrogen and pH for the grape juice analyses. Wine pH and total acidity also differed significantly. / AFRIKAANSE OPSOMMING: In die Mediterreense sones word plantwaterstatus beskou as `n hooffaktor wat groei en produksie van `n wingerdstok negatief beinvloed. In hierdie sones kan wingerdstokke vir lang periodes `n tekort aan water ervaar a.g.v `n tekort aan reënwater gedurende die somer en lae beskikbaarheid van besproeingswater. Die vraag na water vir landbou is ook konstant besig om toe te neem in dié sones en die tendens sal voorduur a.g.v die groei in die wêreldbevolking, die effek van klimaatsveranderig op reënvalpatrone en die hoë verdampingsfaktor. Die wingerd- en wynstreek van die Wes-Kaap word ook geklassifiseer as Mediterreens en wingerdstokke in hierdie streek ervaar dikwels waterspanning wat deur hoë evapotranspirasie en min beskikbare grondwater veroorsaak word. Van die faktore wat die waterstatus van `n wingerdstok bepaal is onder andere die waterpotensiaal van die grondlae rondom die wortelstelsel, die grootte van die wingerdlowerraamwerk en die evapotranspirasiebehoefte. Die omvang van `n wingerdstok se lower binne die prieel word beskou as `n aanduiding van wingerdstokgroeikrag en variasie in groeikrag tussen wingerdstokke is `n algemene verskynsel in die Wes-Kaap. Die rangorde, wat die effek van die verskeie faktore wat groeikragvariasie tussen wingerdstokke bepaal, word steeds gedebatteer. Die debat kan groottendeels toegeskryf word aan die beduidende hoeveelheid variasie tussen wingerde waarmee navorsers te doen kry in wingerdkundige studies. Hoewel, met onlangse vordering aangaande afstandswaarnemingstegnologie is daar nou nuwe metodes beskikbaar om wingerdgroeikrag te evalueer. Dit is kommerwekend om te dink dat `n wyndruifwingerd normaalweg as `n homogene eenheid bestuur word as dit kom by besproeiing. Veral met die wete dat groeikragvariasie tussen wingerde algemeen erken en aangeteken word, en dat volhoubare waterstatus van `n wingerdstok van kardinale belang is vir produksie en kwaliteit van wyndruiwe. Die aanname dat wyndruiwe, die sap- en ook wynkwaliteit homogeen sal wees regdeur `n wingerdblok is egter meer kommerwekkend. Na aanvang van dié denke is daar `n studie geloods om die interaksie tussen wingerdstokgroeikrag en wingerdstokwaterstatus te evalueer. Met die studie is verskeie besproeiingsregimes aangebring binne `n kommersiële wingerd wat interne groeikragvariasie tentoonstel. Groeikragvariasie was geïdentifiseer deur middel van multispektrale lugfotos terwyl die wingerdstok se waterstatus geëvalueer is met behulp van plantgebaseerde metings in die verskillende groeikragareas. Die waterstatus van die grond is geëvalueer tesame met die vegetatiewe groei van die wingerd sodat die groeikragvariasie en die invloed van die plantwaterstatus op die groeikrag bepaal kon word. Die reproduktiewe groei is deurlopend gemonitor voor die effek van wingerdstokwaterstatus en wingerdstokgroeikrag op druifsamestelling en wynkwaliteit bepaal is. Daar was `n goeie ooreenkoms tussen die verskeie metodes wat gebruik is om wingerdgroeikrag te bepaal. Snoeimassa aan die einde van die seisoen was ooreenkomstig met die blaaroppervakte (hooflootblare en sylootblare) wat tydens vegetatiewe groei gemeet is, en het ook goed korreleer, met die multispektrale lugfotos se hoof groeikragklassifikasie. Korrelgewig en -volume het reageer op die verskeie besproeiingsregimes, en daar was `n toename in korrelgewig en -volume saam met die afname in watertekort van een regime tot `n ander. Daar was beduidende verskille tussen die verskeie klassifikasies t.o.v. korrelsamestelling analise en wynevaluasie. Die suikerinhoud per korrel, totale fenole, totale rooi pigment, appelsuur, stikstof en pH het verskil in druiwesap analises. Die pH en suur van die wyne het ook beduidend verskil.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: One of the most striking qualities of wine is its tart, sour taste. The sensory perception of sourness is mainly attributed to the presence of hydrogen ions (protons) at high concentrations. Large amounts of weak carboxylic acids (organic acids) are the main sources of these ions within wine. Once wine enters a person's mouth, the dissociable protons of the weak organic acids within wine are partially neutralized or, in other words, titrated by the saliva secreted inside one's mouth. This explains why the duration and intensity of a wine's sourness is related to its titratable acidity content. The sour taste of wine is usually considered refreshing and it helps balance wine flavour. In fact, wines become watery when its titratable acidity content is too low. After alcoholic fermentation, the titratable acidity of wine will usually be less than that of the grape juice from which was made due to ethanol-induced precipitation of potassium bitartrate crystals and partial consumption of malic acid by fermenting wine yeasts. Occasionally however, increases in titratable acidity are observed during alcoholic fermentation. If wine is produced from grape juice with optimum levels of titratable acidity, unforeseen increases in titratable acidity during alcoholic fermentation can be detrimental to the quality of the final product. Although the net production of malic acid by wine yeasts contributes to increases in titratable acidity seen during grape juice fermentations, the production of succinic acid is regarded as the primary contributor. In fact, succinic acid accounts for approximately 90% of the non-volatile acids produced during fermentation of grape juice. Between 0.5 and 1.5 g/L succinic acid is normally found in wine, but higher concentrations thereof (up to 3.0 g/L) have been detected within certain red wines. Acidity adjustments should preferably be carried out before the onset of alcoholic fermentation to allow better integration of the added compound(s) and to ensure that conditions during fermentation favour the quality and microbial stability of the final product. In doing so unfortunately, winemakers run the risk of ending up with wines that may taste too sour if they are unable to accurately predict and take into consideration the amount of succinic acid produced during alcoholic fermentation. Knowledge with regard to the factors involved in succinic acid's production by fermenting wine yeasts is therefore required in order to manage the titratable acidity of wines more accurately. Ever since Louis Pasteur first noticed succinic acid amongst the by-products of alcoholic fermentation, attempts have been made to determine the metabolic pathways and factors involved in its production by fermenting wine yeasts. Up until now however, it remains unclear why wines sometimes end up with exceptionally high levels of succinic acid. For these reasons it was decided to investigate the possible causes of very high succinic acid concentrations within wine. Due to complexity of grape juice's chemical composition and the problems associated with sterilizing grape juice, fermentation experiments were conducted within a chemically defined grape juice-like medium. Succinic acid production by nine different industrial wine yeast strains was studied under various conditions with regard to the nutrient status of the synthetic grape juice, temperature and availability of molecular oxygen during alcoholic fermentation. The amount of succinic acid produced during alcoholic fermentation was found to depend on the yeast strain, fermentation temperature and chemical composition of the synthetic grape juice. Out of the nine commercial yeast strains selected for this study, strain WE372 produced the largest amount of succinic acid in synthetic grape juice at 28°C. Strain WE372 produced significantly smaller amounts of acetic acid than the other yeast strains of this study and very little acetic acid at 28°C, which indicated that strain WE372 may have less acetaldehyde dehydroganase activity than the other yeast strains of this study under the conditions tested. The effect this has on NAD: NADH balance is the probable cause for its ability to form more glycerol, succinic and malic acid than the other strains. Results from our study show that succinic acid production is influenced primarily by the metabolizable fraction of YAN, which we termed metabolically available nitrogen (MAN). Succinic acid production by fermenting yeasts will be favoured by moderate to high fermentation temperatures (20°C to 28°C) in grape juice with a nicotinic acid and/ or nicotinamide deficiency, high sugar content (200 g/L to 240 g/L), moderate amounts of metabolically available nitrogen (300 ± 50 mg/L MAN), the presence of flavonoids and large supplies of unsaturated long-chain fatty acids. Even higher concentrations of succinic acid were produced when oxygen was made available to fermenting yeasts by aerating the fermenting grape juice. Fermentation temperatures below 18°C, too much metabolizable nitrogen (> 450 mg/L MAN), very high concentrations of fermentable sugar (> 240 g/L), lipid deficiencies and a lack of pantothenic acid, thiamine, biotin or pyridoxine will decrease the amount of succinic acid produced fermenting yeasts. / No Afrikaans summary available.