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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
121

Die geografiese invloede op die wynboubedryf in die Suid-westelike distrikte van die Kaapprovinsie

Theron, H. F. January 1932 (has links)
Thesis (MScAgric)--Stellenbosch University, 1932. / No Abstract Available
122

Overexpression and evaluation of an antimicrobial peptide from Heuchera sanguinea (Hs-AFP1) for inhibition of fungal pathogens in transgenic tabacco

De Beer, Abre 04 1900 (has links)
Thesis (MSc)--University of Stellenbosch, 2002. / ENGLISH ABSTRACT: Seed germination is the most vulnerable time in a plant's life cycle, since the thick protective seed coat ruptures and the moist and humid soil environment not only favours seed germination, but also the growth and development of plant pathogens. Infection of plant seeds during germination, however, is the exception rather than the rule. Plant seeds have - - -developed a--cemplex preformed defense mechanism that includes anttfungal agents thatdiffuse into the surrounding environment to form a protective layer around the seed. This protective layer prevents fungal and bacterial pathogens from infecting the young seedling. Over the last decade, scientists have studied the defense mechanisms of different seeds in an effort to understand and ultimately to introduce and/or manipulate these mechanisms in plants as part of the plant's endogenous disease resistance to pathogens. Various chemical compounds, peptides and proteins that showed strong in vitro activities against various fungi were isolated in these efforts. The mere demonstration of in vitro activity alone, however, is not sufficient to assign a defense role to these antifungal agents. Typically, mutant plants that have lost the ability to produce the antifungal agent, or mutants that are overproducing the agent, have been used to correlate the mutant phenotype to either a decline or increase in disease resistance respectively. Genetic transformation and the subsequent development of transgenic plants have made an unprecedented impact in this regard, specifically in understanding the role of specific defense-related proteins and their interaction with plant pathogens. In this study, the antifungal peptide, Hs-AFP1, from Heuchera sanguinea, a plant defensin, was evaluated in a heterologous in planta environment as a defense protein with potential for engineering disease resistant crops. The in vitro assays performed with Hs-AFP1 against Botrytis cinerea showed antifungal activities of 88% growth inhibition at a concentration of 8 J,lg/ml of the purified peptide, while inducing a characteristic hyperbranching effect on the Botrytis hyphae. Tobacco was subsequently transformed with a construct, pFAJ3068, expressing Hs-AFP1 under the strong constitutive 35S promoter. The peptide was targeted to the apoplastic region with the signal peptide from Mj-AMP2, an antimicrobial peptide from Mirabilis jalapa. Due to reports of peptide instability in transgenic plant systems, two additional constructs were prepared and transformed into tobacco to anticipate possible Hs-AFP1 instability in the heterologous tobacco environment. A putative peptide stabilization construct, pHs-EXG1, consisted of a fusion between Hs-AFP1 and the antifungal exo-glucanase (encoded by EXG1) from Saccharomyces cerevisiae. A control construct, pMj-EXG1, expressing EXG1 targeted to the apoplastic region with the Mj-AMP2 signal peptide, was also prepared and transformed into tobacco to normalize the background antifungal activity as a result of the exoglucanase in the fusion construct lines. Tobacco was successfully transformed with pFAJ3068, pHs-EXG1 and pMj-EXG1, resulting in transgenic tobacco lines designated THs, THE and TME respectively. Transgene expression was confirmed for the THs and THE transgenic lines. The translation of these transcripts into proteins was also confirmed with Western blot analysis. Moreover, the heterologous production of Hs-AFP1 in tobacco led to an increase in disease resistance to B. cinerea in the THs lines in comparison with the untransformed tobacco controls. An increase of up to 42% in disease resistance was observed in an in planta detached leaf assay. Crude protein extracts from the THs lines were also analyzed in an in vitro quantitative fungal growth assay. This assay confirmed the results obtained with the disease resistance assay, with crude protein extracts exhibiting up to 40% fungal growth inhibition. The incubation of B. cinerea in the presence of crude protein extracts from THs lines resulted in hyperbranching of the fungal hyphae, which is characteristic of Hs-AFP1 activity. From these analyses it was clear that the heterologously expressed Hs-AFP1 was quite stable in the transgenic environment. The fusion between Hs-AFP1 and EXG1 did not increase the stability of Hs-AFP1, but rather led to a loss of the Hs-AFP1 activity. All the analyses performed showed the THE lines to be reduced in their ability to inhibit fungal infection in comparison to the THs line. Also, microscopic analysis of the effects of the crude THE extracts on B. cinerea growth showed no hyperbranching activity, again confirming the loss of peptide activity due to the fusion to EXG1. This is in agreement with previous work, in which sarcotoxin 1A was fused to a reporter gene and also lost activity. Although integration of the Mj-EXG1 expression cassette was confirmed, no mRNA levels could be detected with Northern blot or RT-PCR analysis of the TME lines. These lines also did not show any in vitro antifungal activities, probably indicating post-transcriptional gene silencing. This silencing was overcome in the fusion constructs that were expressed in the THE plant lines. These lines also showed EXG1 protein activity, as measured by ~-glucosidase assays. Although the THE lines did not serve the functions originally envisaged, they fortuitously showed that a fusion strategy might stabilize glucanase expression in a transgenic environment. A variety of glucanases have been shown to be prone to gene silencing when overexpressed in a plant environment and the yeast glucanase can now be added to that list if it is not present as a fusion protein. Overall, this study confirmed that Hs-AFP1 is involved in plant defense systems and provided valuable information on the stability of small peptides in a heterologous environment. The positive results obtained with overexpressed Hs-AFP1 on fungal inhibition in this study merits further investigations into the use of this peptide in the engineering of disease-resistant crops. / AFRIKAANSE OPSOMMING: Saadontkieming is die mees vatbare tyd vir siekteontwikkeling gedurende 'n plant se lewenssiklus. Die saadhuid bars en die vogtige grondkondisies bevoordeel nie net saadontkieming nie, maar ook die groei en ontwikkeling van plantpatogene. Infeksie van plantsade tydens ontkieming is egter die uitsondering eerder as die reël. Plantsade besit komplekse -veraeaigingsfueganfsmes-reen moontlike - patoqeeninteksies. Die meqanismes sluit die produksie van antifungiese agense, wat tydens saadontkieming na die omliggende omgewing diffundeer om 'n beskermende sone om die ontkiemende saad te vorm, in. Die gevolglike antifungiese sone beskerm die saad teen infeksie deur bakterieë en swamme. Gedurende die laaste dekade het navorsers baie aandag aan die bestudering van plantsaadverdedigingsmeganismes gegee. Dié kennis word gebruik om die verdedigingsmeganismes beter te verstaan, asook om dié meganismes te manipuleer en/of oor te dra aan plantspesies met inherente swak weerstandsmeganismes wat gereeld aan plantpatogeeninfeksies onderhewig is. Navorsing op plantsade het tot die isolasie van verskeie chemiese agense, peptiede en proteïene, wat sterk in vitro aktiwiteite teen 'n wye reeks swampatogene vertoon, gelei. Die vermoë van dié agense om swamme in 'n in vitro omgewing te inhibeer, is alleen egter nie 'n bewys dat hulle 'n rol in plantverdeging speel nie. Studies waar mutante gebruik word, is gewens om addisionele bewys te lewer dat die substanse 'n rol in plantverdediging vervul. Sodanige mutante sluit plantlyne, waarin die geen van belang gemuteer is of ooruitgedruk word om so die rol van die geen in 'n in planta omgewing te bepaal in. In hierdie toepassings het genetiese transformasie en die daarstelling van transgeniese plante 'n ongeëwenaarde bydrae gelewer. In dié studie is die antifungiese peptied, Hs-AFP1, wat aan die peptiedgroep van plant- "defensins" behoort en van Heuchera sanguine a afkomstig is, in 'n heteroloë in planta omgewing geëvalueer as 'n verdedigingspeptied met die potensiaal om in die generering van transgeniese siektebestande gewasse gebruik te word. Die antifungiese aktiwiteit van Hs-AFP1 is teen Botrytis cinerea in 'n in vitro reaksie geëvalueer, waar die toediening van 8 ,",g/mlgesuiwerde Hs-AFP1 peptied aanleiding gegee het tot 'n 88% afname in hifegroei van B. cinerea. Hipervertakkings van swamhifes, 'n kenmerkende eienskap van Hs-AFP1 aktiwiteit, kon duidelik waargeneem word. Tabakplante is voorts getransformeer met 'n konstruk, pFAJ3068, wat die koderende geen van Hs-AFP1 onder die sterk konstitutiewe CaMV 35S promotor bevat het. Die peptied is met behulp van die seinpeptied wat afkomstig is van die Mirabilis jalapa antimikrobiese peptied, Mj-AMP2, na die apoplastiese omgewing geteiken. Voorheen is gerapporteer dat transgeniese peptiede in die heteroloë omgewing soms onstabiel is. Dit het gelei tot die generering van twee addisionele konstrukte om die moontlikheid van peptiedonstabiliteit te ondervang. 'n Stabiliseringskonstruk, pHs-EXG1, bestaande uit In versmelting tussen Hs-AFP1 en In antifungiese eksoglukanase van Saccharomyces cerevisiae, gekodeer deur EXG1, is in tabakplante getransformeer. In Kontrolekonstruk, pMj-EXG1, met die EXG1-geen saam met die Mj-AMP2-seinpeptied, is ook voorberei en in tabakplante getransformeer. Dit is gebruik om die antifungiese aktiwiteit van die eksoglukanase in die antifungiese aktiwiteitstoetse van die stabiliseringskonstruk te kwantifiseer en te normaliseer. Tabak is suksesvol met pFAJ3068, pHs-EXG1 en pMj-EXG1 getransformeer, wat onderskeidelik gelei het tot die sogenaamde THs, THE en TME transgeniese tabaklyne. Transgeentranskripsie en -translasie in die THs en THE tabaklyne is onderskeidelik deur Noordelike- en Westelike-kladanalises bevestig. Die aktiewe uitdrukking van Hs-AFP1 het die vermoë van tabakplante om B. cinerea infeksies te weerstaan, met tot 42% verhoog in vergelyking met ongetransformeerde kontrole tabakplante tydens 'n in planta siekteweerstandstoets. Totale proteïenekstrakte van THs tabaklyne is voorts ook in In in vitro inhibisietoets geëvalueer, wat gelei het tot resultate wat goed met dié van die in planta toetse ooreenstem. Die totale proteïenekstrakte het swamgroei met 40% geïnhibeer en die kenmerkende hipervertakking van Hs-AFP1-aktiwiteit is ook mikroskopies waargeneem. Resultate wat verkry is vanaf al die analises wat op die transgeniese THs tabaklyne uitgevoer is, het aangedui dat Hs-AFP1 baie stabiel in die heteroloë tabakomgewing is en peptiedstabiliteit was dus nie In probleem, soos verwag is nie. Die fusie tussen Hs-AFP1 en EXG1 het dus nie die stabiliteit van die reeds stabiele Hs-AFP1 peptied verder verbeter nie, maar het wel tot die verlies van Hs-AFP1 aktiwiteit gelei. Die antifungiese analises van die THE tabaklyne het verder bevestig dat dié lyne selfs swakker inhibisie van B. cinereainfeksies tot gevolg gehad het, as ongetransformeerde tabakplante. Mikroskopiese analises van totale THE proteïenekstrakte het voorts ook geen kenmerkende hipervertakkings in die swamhifes vertoon nie, wat alles daarop dui dat die Hs-AFP1-deel van die fusieproteïen as gevolg van die fusie met EXG1 geïnaktiveer is. Dié resultaat is in lyn met vorige navorsing, wat getoon het dat In ander peptied, sarcotoxin 1A, sy antifungiese aktiwiteit verloor indien dit met In verklikkergeen versmelt word. Alhoewel integrasie van die pMj-EXG1-konstruk in die TME-tabaklyne bevestig is, kon geen mRNA met Noordelike-klad- of trutranskriptase-PKR (RT-PKR)-analises waargeneem word nie. Die TME plant het ook geen antifungiese aktiwiteit in in vitro toetse getoon nie en dit het geblyk dat die pMj-EXG1-konstruk aan geenafskakeling in die heteroloë tabakomgewing onderworpe was. Dié afskakelingseffek is egter in die THE plante oorkom, aangesien laasgenoemde sterk EXG1 proteïenaktiwiteit met J3-glukosidase aktiwiteitstoetse vertoon het. Alhoewel die THE plante nie die stabiliteit van Hs-AFP1 verbeter het nie, het dit onwerwags tot die stabilisering van EXG1 in In heteroloë omgewing gelei. Versmeltingstegnologie kan dus moontlik gebruik word as 'n strategie om ander glukanases, wat bekend is vir geenafskakeling in transgeniese omgewings, heteroloog uit te druk. In die geheel gesien, het dié studie getoon dat Hs-AFP1 'n onbetwiste rol in plantverdedigingsmeganismes speel en daar is voorts ook meer kennis oor die stabiliteit van peptiede in 'n heteraloë plantomgewing ingewin. Die positiewe resultate t.o.v. die verhoogde siekteweerstand in die transgeniese THs plantlyne regverdig ook die verdere bestudering van dié peptied om transgeniese siekteweerstand in gewasse te bewerkstellig.
123

Consumer acceptance of a selection of South African red wines : intrinsic, extrinsic and socio-demographic influences

Basson, Shantelle 03 1900 (has links)
Thesis (MSc Food Sc)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: In this study an industry-selected and diverse range of South African red wines were analysed for sensory and chemical attributes, as well as degree of liking using a target group of black South African consumers. Segments of consumers that differed in degree of liking were then tested for their response to intrinsic (sensory) and extrinsic (non-sensory) cues. The selection of wines included eighteen dry and natural sweet red wines, representing low-end inexpensive wines together with high-end, top quality wines. Sensory profiles for all samples were established using Quantitative descriptive analysis (QDA). The results revealed that cultivar specific dry red wines associated with a wide range of sensory descriptors such as woody, vegetative and fruity, while the sweet red wines associated with the fruity and sweet-associated attributes. Chemically there was a significant variation between wines regarding the alcohol and sugar content. Gas chromatography with flame ionisation detection (GC-FID) indicated the major volatile constituents present in the wine, i.e. esters, alcohols and fatty acids. When investigating the association between the chemical and sensory data, it was revealed that the red blends were driven by the presence of alcohols and esters, and sensory descriptors such as high roast oak, coffee and mixed spice, whereas the red cultivar wines were mostly driven by fatty acids and esters and the sensory descriptors, green bean and asparagus. The sweet red blends were closely associated with acids and the sensory descriptors sweet-associated and floral. Degree of liking of a subset of 18 wines was investigated based on the preferences of black consumers from the Western Cape area, South Africa. These consumers predominantly preferred the sweet red wines with high sugar content, in a blind tasting session. Purchase intent was also evaluated by viewing actual photographs of packaging formats of the respective wines and the results indicated that the consumers preferred the well-known cultivar wines with a perception of value and style. Cluster analysis was furthermore performed to ascertain whether these consumers differed in their degree of liking of the intrinsic character of the respective wines. Four different clusters of consumers were identified: 1) Consumers preferring both dry and sweet red wines equally, 2) Consumers who strongly favoured sweet red wines and moderately liked dry red wines, 3) Consumers who strongly favoured sweet red wines with little preference for dry red wines; and 4) Consumers preferring dry red wines. Consumers were also probed on their general opinions or perceptions on the extrinsic character of the wines, and thus factors that influence the purchasing process. It was found that black consumers who don‟t consume wine often, preferred wines that they are familiar with, while consumers that drink wine more frequently enjoy to broaden their horizons by experimenting with more expensive wine brands. Extrinsic or non-sensory cues such as alcohol content, label, vintage, price and cultivar were found to be the most important considered factors when purchasing red wines, while awards and type of closure were regarded as the least important. It was also found that the discerning consumers, who purchase high-end wines, took more of the latter aspects into consideration, whereas consumers who purchase low-end wines considered a limited number of the non-sensory cues. / AFRIKAANSE OPSOMMING: In hierdie studie is 'n diverse reeks industrie-geselekteerde, Suid-Afrikaanse rooiwyne geanaliseer vir hul sensoriese en chemiese eienskappe. Verbruikersvoorkeur van die wyne is getoets, asook tot watter mate verbruikersvoorkeure beïnvloed word deur intrinsieke (sensoriese) en ekstrinsieke (nie-sensoriese) faktore. Die reeks van agtien wyne het bestaan uit droë en soet rooi wyne, wat op hul beurt verder verdeel kan word in goedkoper, kwaliteit wyne en duurder, ultra-premium wyne. Die sensoriese profiel van al die wyne is bepaal deur beskrywende sensoriese analise. Resultate het getoon dat die kultivar-spesifieke droë rooiwyne geassosieer word met 'n wye reeks sensoriese eienskappe soos houtagtig, kruidagtig en vrugtig, terwyl die soet rooiwyne beskryf is as vrugtige en soet-geassosieerd. In terme van die chemiese analises was daar betekenisvolle verskille betreffende die alkohol- en suikerinhoud van die wyne. Gas chromatografie gekoppel met vlam-ioniserende deteksie (GC-FID) het die mees vlugtige verbindings teenwoordig in die wyn aangedui, naamlik esters, alkohole en vetsure. Met die korrelasie van die chemiese en sensoriese data is gevind dat die droë versnitwyne gedryf word deur die teenwoordigheid van alkohole en esters, asook sensoriese eienskappe soos gehout, koffie, en gemengde spesery, terwyl die kultivar-spesieke wyne weer meestal gedryf word deur vetsure en esters en sensoriese eienskappe soos groenboontjie en aspersie. Die soet rooiwyne het chemies geassosieer met sure en sensoriese terme soos soet-geassosieerd en blomagtig. Die aanvaarbaarheid van 'n kleiner groepering wyne is bepaal deur gebruik te maak van swart verbruikers in die Wes-Kaap area, Suid-Afrika. Die verbruikers het in 'n blinde proesessie onderskeie wyne se wynverpakking besigtig en aangedui of hulle die wyne sou koop. Hierdie resultate het getoon dat die verbruikers bekende kultivarwyne verkies wat 'n persepsie van waarde en styl geïllustreer het. Segmentasie tegnieke is op die data uitgevoer ten einde te bepaal of verbruikers in groepe verdeel kan word, wat betref hul voorkeur van die sensoriese of intrinsieke eienskappe van die wyne. Vier verskillende groepe is geïdentifiseer, nl. verbruikers wat 1) droë en soet rooiwyne ewe veel verkies; 2) soet rooiwyne en tot 'n mate ook droë rooiwyne verkies; 3) soet rooiwyne en tot 'n mindere mate droë rooiwyne verkies; en laastens 4) slegs droë rooiwyne verkies. Verbruikers se algemene opinies en persepsies betreffende die ekstrinsieke eienskappe van die wyne is ook ondersoek, met ander woorde faktore wat die aankoop van wyne beïnvloed. Daar is gevind dat swart verbruikers wat nie gereeld wyn drink, bekende handelsmerke verkies, terwyl verbruikers wat gereeld wyn drink, daarvan hou om hul horisonne te verbreed en te eksperimenteer met 'n verskeidenheid handelsmerke. Ekstrinsieke of nie-sensoriese aspekte soos, alkohol-inhoud, etiket, oesjaar, prys en kultivar is die belangrikste faktore wat in ag geneem word wanneer rooiwyne gekoop word, terwyl wyntoekennings en die feit dat die wyn met kurke gebotteleer word, nie as belangrik beskou word nie. Daar is ook gevind dat die meer ingeligte verbruiker, wat hoë kwaliteit wyne koop, meer van die bogenoemde aspekte in ag neem tydens die aankoopproses, terwyl die verbruiker wat meer geneig is om goedkoper wyne te koop, slegs 'n paar ekstrinsieke faktore in ag neem.

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