Large public events, such as concerts, rallies, and festivals, impact turf health when held on natural turfgrass surfaces. The impact associated with these events is due to the placement of physical structures such as stages and seating areas and pedestrian and vehicular traffic on the turf surface. Trafficked turf protection covers, which are field covers meant to be placed directly on the turf surface where pedestrian or vehicular traffic is expected and/or equipment will be placed, can be used to minimize damage to the turf surface. Scientific data on turf response to these covers is lacking. Four cover treatments comprised of a non-covered non-trafficked control, plywood, plywood + Enkamat Plus, and white high-density polypropylene [single sided (Terratile) or double sided (Matrax)] were applied to tall fescue (Festuca arundinacea Schreb.) and effects of light intensity, duration of covering, season and soil moisture were evaluated. Growth chambers and field experiments were conducted in 2010-2011. Tissue samples were taken in growth chambers experiments every four days over the 20-day period to analyze chlorophyll (Chl a, Chl b, Chl a+b) and carotenoids (carot) under split factors of light intensity (12hr, PAR 530 μmol m-2 s-1, 5 μmol m-2 s-1) and soil moisture (50%, 75% of pot soil moisture capacity). Field trial treatment effects were observed every two days and eight days after cover removal in the spring, summer and fall and a normalized difference vegetative index (NDVI) measure was used at the conclusion of each trial period to confirm visual ratings. Covers that allowed light transmission to the canopy provided the best visual retention of percent green cover and higher contents of Chl a, Chl b, Chl a+b and carot. However, when treatments were tested under conditions that simulated low light under a concert stage (PAR 5 μmol m-2 s-1), covers performed similarly. Moderate soil moisture increased Chl b and carot content under covers. Field trials showed that plywood and plywood + Enkamat allowed for acceptable covering periods of six days in spring, four days in fall, and zero days in summer. Summer conditions shortened the number of days (8 -10) thattall fescue could be covered with Matrax and Terratile and still maintain an acceptable level of green cover. Matrax performed the best during high temperatures and did not tend to sink into the turf in saturated soil. All covers exhibited desirable qualities and limitations that should be considered for turf protection during an event. / Master of Science

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Cuscuta campestris is an obligate parasitic plant that lacks expanded leaves and roots and requires a host to complete its lifecycle. Parasite-host connections occur via an haustorium, a unique organ that acts as a bridge for the exchange of water, nutrients, macromolecules like mRNA, microRNA, and proteins, and microorganisms. Studies of Cuscuta spp. are challenging due to its dependence on the host and other host influences on the parasite. Recent research has shown intriguing aspects of Cuscuta biology like exchange genetic material with its hosts and loss of genes involved in processes such as high photosynthetic rates and defense. We developed new tools and methodologies that allow us to explore C. campestris biology in an unprecedent way. Foremost of these is an axenic method to grow C. campestris on an Artificial Host System (AHS). The AHS allows C. campestris to display its entire life cycle in vitro, including seed production. Using the AHS, we studied haustorial function, determining the role of nutrients and phytohormones on parasite haustorium development and growth, and found genes involved in haustorial function. The AHS allowed us to demonstrate the positive effect of light on C. campestris growth in the absence of a photosynthetic host and to investigate carotenoid- and ABA- related processes in the haustorial regions. We also wanted to understand how C. campestris defenses work independently from a plant host, so we studied the parasite responses to the bacterial epitope flg22 and the bacteria Peudomonas syringe. Our findings indicate that C. campestris is able to sense flg22, but its response differs from those observed in other non-parasite plants. Transcriptomic analysis revealed up-regulation of genes related to biotic and abiotic stresses, and downregulation of genes related to cuticle development. Our study contributes to understanding the C. campestris immune response in the absence of a host plant. Taken together, this research contributes novel methodologies that enable insights into C. campestris biology without the interference of a plant host on the parasite. / Doctor of Philosophy / Field dodder (Cuscuta campestris) is a parasitic plant that lacks leaves and roots and attacks a wide range of plants, such as tomato and beets. Dodders are not able to carry out full photosynthesis and thus are incapable of producing enough food or obtaining water to survive on their own, so they parasitize other plants. Dodders have developed specialized structures called haustoria that allow them to take resources directly from their hosts. Studying dodder is challenging due to the dependence of the parasite on its host, such that effects of one plant on the other are hard to disentangle. We developed new tools and methodologies that allow us to explore the biology of dodder in unprecedent ways. We developed an Artificial Host System (AHS) that allows the growth and study of dodder without involving a living host plant. Thanks to this new tool, we were able to improve understanding of the function of the haustorium, discover nutrients and growth factors that are indispensable for dodder development, and prove that dodder growth benefits from light. Using the AHS, we compared haustorial regions and shoot tips of dodder to identify genes specific to haustorial function. Additionally, we studied the responses of dodder to bacteria to understand how it reacts to microbial colonization. Our studies contribute with the development of novel methodologies that allow unprecedent discoveries into the biology of dodder. We expect that this work will promote the study of parasitic plant biology.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / A deficiÃncia de vitamina A (DVA) à considerada um importante problema de saÃde pÃblica em todo o mundo e o papel da DVA na integridade epitelial intestinal ainda nÃo està bem compreendido. O objetivo do trabalho foi investigar a associaÃÃo do retinol (vitamina A) e carotenÃides com a barreira funcional intestinal em crianÃas de uma comunidade urbana em Fortaleza, no Nordeste do Brasil. Participaram do estudo 102 crianÃas de 2 meses a 9 anos de idade. O estado nutricional das crianÃas foi avaliado atravÃs do escore âzâ de peso por altura (WHZ), mostrando que 19,6% (20/102) das crianÃas apresentaram desnutriÃÃo leve (âzâ escore de -1 a -2). Todas as crianÃas tiveram as concentraÃÃes de retinol determinadas e nenhuma estava severamente deficiente (< 0,35 ÂM); 2,9% (3/102) das crianÃas apresentaram nÃveis moderados de deficiÃncia de vitamina A (0,36 - 0,70 ÂM); 20,6% (21/102) apresentaram leve deficiÃncia (0,71 - 1,05 ÂM) e 76,5% (78/102) apresentaram concentraÃÃes suficientes de retinol no sangue (> 1,05 ÂM). A razÃo de lactulose/manitol (L/M) estava anormal em 49% (47/97) das crianÃas quando comparadas com crianÃas saudÃveis de uma mesma regiÃo geogrÃfica. Os carotenÃides, luteÃna, &#946;-criptoxantina e beta-caroteno no soro das crianÃas do Parque UniversitÃrio, apresentaram correlaÃÃes inversas significativas com a razÃo de L/M. As proteÃnas de fase aguda (proteÃna C reativa, CRP e glicoproteÃna &#945;1-Ãcida, AGP) apresentaram correlaÃÃes inversas com retinol. O retinol correlacionou-se significativamente com a proteÃna ligadora de retinol (RBP) e com a transtiretina (TTR). Esses dados sugerem que a alteraÃÃo na barreira funcional intestinal provavelmente ocorreu atravÃs do caminho paracelular com baixas concentraÃÃes de carotenÃides no soro. Os carotenÃides, precursores do retinol, poderiam assim promover um melhor marcador para a barreira funcional intestinal alterada do que as medidas de RBP ou retinol. As concentraÃÃes sanguÃneas de retinol correlacionaram-se com a RBP e TTR e correlacionaram-se inversamente com a AGP e com a CPR / Vitamin A deficiency (VAD) is considered an important public health problem worldwide and the role of VAD on intestinal epithelial integrity is not well understood. To investigate the association of retinol (vitamin A) and carotenoids with the intestinal barrier function from children in an urban community in Fortaleza, Northeastern Brazil, one hundred and two children from 2 months to 9 years old participated in the study. The nutritional status of these children, measured by weight for height z-score (WHZ), showed that 19.6% (20/102) of them had mild malnutrition (-1 to â2 z-score). All children had their serum retinol concentration measured and none were severely deficient (<= 0.35 ÂM), 2.9% (3/102) were moderately (0.36 â 0.70 ÂM), 20.6% (21/102) were mildly (0.71 â 1.05 ÂM) deficiencies; 76.5% (78/102) were vitamin A sufficient (> 1.05 ÂM). The lactulose:mannitol (L/M) ratio was abnormal in 49% (47/97) of children when compared to healthy children in the same geographic area. Serum carotenoids, lutein, beta-cryptoxanthin and beta-carotene showed significant inverse correlation with the L/M ratio. Acute phase proteins (C-reactive protein; CRP; and &#945;-acid glycoprotein; AGP) were significantly inversely correlated with retinol. Retinol was significantly correlated with retinol-binding protein (RBP), and with transthyretin (TTR). These data suggest a disruption of intestinal barrier function due to paracellular pathway with low serum concentrations of carotenoids. The retinol precursors, carotenoids, may provide a better marker for disrupted intestinal barrier function than measurements of RBP or retinol. Serum retinol concentrations correlate with RBP and TTR and inversely correlate with AGP and CRP

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: Previous research has shown that carotenoids are precursors of C13-norisoprenoid aroma compounds in wine. C13-norisoprenoids have low threshold values in wine with the most prominent C13-norisoprenoids being β-damascanone and β-ionone which contribute honey and floral like aroma to wine. Chlorophyll and its derivates have also been detected in wine with potential to be precursors to aroma compounds. Apart from the contribution of these pigments to wine aroma and quality they are vital role players in photosynthesis and are widely found in plants and plant products. The main functions of these pigments in plants are light collection and light-protection. Research has shown that environmental conditions, climate, light exposure of bunches and soil water deficit influence the carotenoid content of grape berries. Furthermore the concentration of carotenoids and chlorophylls has also been shown to differ between cultivars. No research in this regard has been done on Merlot grape berries. With this in mind, the aim of this study was to evaluate the effect of vigour and soil water content on the evolution of carotenoids and chlorophylls through ripening of grape berries from the cv. Merlot. However, when looking at methods to analyse carotenoids and chlorophylls in berry tissue, especially lyophilised tissue, there were no readily available methods. Thus, an extraction method to identify and quantify the carotenoid and chlorophyll profile of lyophilised tissue from unripe (green) to ripe (red) Merlot grape berries was needed. In this study the RPHPLC method of Taylor et al. (2006) for carotenoids and the extraction method of Mendes-Pinto et al. (2004) were adapted to analyse both carotenoids and chlorophylls in lyophilised grape tissue. The RP-HPLC method baseline separated all the carotenoids and chlorophylls and their derivatives. Recovery of standards from mock extractions was high, indicating that the extraction procedure was acceptable. However, extraction recovery tested in the matrix of the grape tissue showed less promising results due to the high acid content of grape tissue. Violaxanthin, neoxanthin and the chlorophylls were especially sensitive to low pH conditions which facilitated their degradation. The degradation products of these compounds under acidic conditions were identified as pheophytin a, b, chlorophillide a, pyropheophytin b, cisviolaxanthin, cis-neoxanthin, neochrome, mutatoxanthin and luteoxanthin. There is a possibility that some degradation products were already present in the tissue due to lyophilisation (since the water in the berry was then removed and the acid concentrated). More work is needed to investigate the effect of lyophilisation and storage on the composition of grape tissue of different maturity. The extraction method for grape berry tissue at different ripening stages should also be optimised further to effectively neutralise tissue acidity, without compromising the extraction of carotenoids significantly, in especially green berry tissue. The question as to whether cisisomers and chlorophyll degradation products are naturally present in grape berries or are formed during sampling and processing remains unanswered in the current study. This study confirmed that in general carotenoids and chlorophylls decrease on a per berry (μg/berry) and concentration (μg/g) basis from veraison to harvest. Furthermore, this study was inconclusive in showing that vigour differences have an effect on the rate of synthesis/degradation of carotenoids, chlorophyll and some other ripening parameters, namely malic acid, total glucose and fructose, total tannin and total anthocyanin, from pre-veraison (pea size) to harvest. Additionally, no significant effect of soil water content on carotenoids, chlorophylls and ripeness parameters was found in this study, most likely due the fact that high soil water capacity was found in lower soil layers which may have prevented significant differences in grapevine water status. Experimental plots selected for vigour differences based on normalised difference vegetation index (NDVI) images, pruning mass and soil water measurements by means of a neutron probe, showed significant differences in soil water content in only the first 30 cm of the soil for the ripening seasons studied. Predawn plant water potential measurements, however, indicated that none of the experimental vines experienced severe water stress which was previously shown to effect carotenoid content of grapes. The carotenoid 5,8-epoxy--carotene was quantified for the first time in grapes and represents a significant amount of the total carotenoids present at harvest. All the carotenoids and chlorophylls except -carotene appeared to be sensitive to seasonal variation in climatic conditions. Lutein and β-carotene were found to be the most abundant carotenoids present in Merlot grape berries together with chlorophyll a for both seasons studied. The values of these carotenoids also correlated well with previous research. However, chlorophyll a was found in much larger quantities in Merlot berries compared to reported data. This is possibly because in this study the chlorophyll degradation products were included in the calculation of chlorophyll a. Multivariate analysis showed promising preliminary prediction models (with correlation values of above 0.8 for both seasons analysed) for the prediction of the concentration of ripeness parameters (glucose, fructose, malic acid, total tannins and anthocyanins) with carotenoid and chlorophyll content. This result highlights the opportunity for the development of a rapid non-destructive method to measure carotenoids and chlorophylls in berries which in turn can predict optimal ripeness. Furthermore, since carotenoids are the precursors to C13- norisoprenoid aroma compounds in wine a preview of the potential contribution of these aromas to wine might be evaluated. Further research is necessary to investigate the possibility of building and validating such models. / AFRIKAANSE OPSOMMING: Vorige navorsing het getoon dat karotenoïede die voorlopers is van C13-norisoprenoïed aromaverbindings in wyn. C13-norisoprenoïede het lae drempelwaardes in wyn, met β- damassenoon en β-jonoon as die prominentste C13-norisoprenoïede wat ‘n bydrae tot die heuning en blomagtige aroma van die wyn maak. Chlorofil en sy derivate is ook reeds in wyn bespeur, met die potensiaal om voorlopers van aromaverbindings te wees. Buiten die bydrae van hierdie pigmente tot wynaroma en -kwaliteit is hulle ook belangrike rolspelers in fotosintese en kom hulle wydverspreid in plante en plantprodukte voor. Die vernaamste funksies van hierdie pigmente in plante is om lig te versamel en om as beskerming teen lig op te tree. Navorsing het getoon dat omgewingstoestande, klimaat, ligblootstelling van die trosse en grondwatertekorte die karotenoïedinhoud van druiwekorrels beïnvloed. Verder is ook getoon dat die konsentrasie van karotenoïede en chlorofille tussen kultivars verskil. Geen navorsing is al in hierdie opsig op Merlot-druiwekorrels gedoen nie. Met hierdie aspek in gedagte was die doelwit van hierdie studie om die effek van groeikrag en grondwaterinhoud op die evolusie van karotenoïede en chlorofille tydens die rypwording van druiwekorrels van die cv. Merlot te evalueer. Wanneer mens egter kyk na die metodes waarvolgens die karotenoïede en chlorofille in korrelweefsel geanaliseer word, is daar geen geredelik beskikbare metodes nie. ‘n Ekstraksiemetode om die karotenoïed- en chlorofilprofiel van geliofiliseerde weefsel van onryp (groen) tot ryp (rooi) Merlot-bessies te identifiseer en kwantifiseer was dus nodig. In hierdie studie is die RP-HPLC metode van Taylor et al. (2006) vir karotenoïede en die ekstraksiemetode van Mendes-Pinto et al. (2004) aangepas om beide karotenoïede en chlorofille in geliofiliseerde druiweweefsel te analiseer. Die basislyn van die RP-HPLC metode het all karotenoïede en chlorofille en hul derivate geskei. Herwinning van die standaarde vanaf skynekstraksies was hoog, wat aandui dat die ekstraksieprosedure aanvaarbaar was. Ekstraksieherwinning wat in die matriks van die druiweweefsel getoets is, het egter minder belowende resultate getoon as gevolg van die hoë suurinhoud van die druifweefsel. Violaxantien, neoxantien en die chlorofille was veral sensitief vir toestande van lae pH, wat hulle afbreking gefasiliteer het. Die afbrekingsprodukte van hierdie verbindings onder suurtoestande is geïdentifiseer as feofitien a en b, chlorofillied a, pirofeofitien b, cis-violaxantien, cis-neoxantien, neochroom, mutatoxantien en luteoxantien. Daar is ‘n moontlikheid dat sommige afbreekprodukte reeds in die weefsel teenwoordig was as gevolg van liofilisering (aangesien die water in die korrel reeds verwyder was en die suur gekonsentreerd was). Meer werk is nodig om die effek van liofilisering en berging op die samestelling van druifweefsel van verskillende rypheid te bepaal. Die ekstraksiemetode vir druifkorrelweefsel op verskillende stadia van rypwording moet ook verder geoptimaliseer word om weefselsuurheid doeltreffend te neutraliseer, sonder om die ekstraksie van karotenoïede noemenswaardig te kompromitteer, veral in groen korrelweefsel. Die vraag of cis-isomere en chlorofil afbreekprodukte natuurlik in die druifkorrels teenwoordig is en of hulle tydens monsterneming en prosessering gevorm word, kon nie in hierdie studie beantwoord word nie. Hierdie studie het bevestig dat karotenoïede en chlorofille oor die algemeen op ‘n korrel (μg/korrel) en konsentrasie (μg/g) basis afneem vanaf deurslaan tot oes. Hierdie studie het nie daarin geslaag om te toon dat groeikragverskille vanaf voor-deurslaan (ertjiekorrelgrootte) tot oes ‘n effek het op die tempo van sintese/afbreking van karotenoïede, chlorofil en ander rypwordingsparameters nie, naamlik op appelsuur, totale glukose en fruktose, totale tannien en totale antosianien. Daar is ook in hierdie studie geen noemenswaardige effek van grondwaterinhoud op karotenoïede, chlorofille en rypheidsparameters gevind nie, heel moontlik as gevolg van die feit dat hoë grondwaterkapasiteit in die laer grondlae gevind is, wat betekenisvolle verskille in wingerdwaterstatus kon verhoed het. Eksperimentele persele wat gekies is vir groeikragverskille op grond van genormaliseerde verskil plantegroei indeks (NDVI) beelde, snoeimassa en grondwatermetings met ‘n neutronvogmeter het net in die eerste 30 cm van die grond noemenswaardige verskille in grondwaterinhoud getoon vir die rypwordingseisoene wat bestudeer is. Voor-sonopkoms plantwaterpotensiaalmetings het egter aangedui dat geen van die eksperimentele wingerdstokke ernstige waterstres ervaar het nie. Sulke stres is voorheen aangedui om ‘n effek op die karotenoïedinhoud van druiwe te hê. Die karotenoïed 5,8-epoksi--karoteen is vir die eerste keer in druiwe gekwantifiseer en verteenwoordig ‘n noemenswaardige hoeveelheid van die totale karotenoïede wat met oes teenwoordig is. Al die karotenoïede en chlorofille behalwe -karoteen blyk sensitief vir seisoenale verskille in klimaatstoestande te wees. Luteïen en β-karoteen was die volopste karotenoïede in die Merlot-druifkorrels, tesame met chlorofil a, vir beide seisoene wat bestudeer is. Die waardes van hierdie karotenoïede was ook goed gekorreleer met vorige navorsing. Chlorofil a is egter in baie groter hoeveelhede in Merlot-korrels gevind in vergelyking met dít wat in die data gerapporteer is. Die rede hiervoor is moontlik dat die chlorofil-afbreekprodukte in hierdie studie in die berekening van chlorofil a ingesluit is. Meerveranderlikeontleding het belowende voorlopige voorspellingsmodelle getoon (met korrelasiewaardes van meer as 0.8 vir beide die seisoene wat geanaliseer is) vir die voorspelling van die konsentrasie van rypheidsparameters (glukose, fruktose, appelsuur, totale tanniene en antosianiene) met karotenoïed- en chlorofilinhoud. Hierdie resultaat beklemtoon die geleentheid vir die ontwikkeling van ‘n vinnige, nie-destruktiewe metode om karotenoïede en chlorofille in korrels te meet, wat op sy beurt optimate rypheid kan voorspel. Aangesien karotenoïede die voorlopers van C13-norisoprenoïed aromaverbindings in wyn is, kan ‘n voorskou van die potensiële bydrae van hierdie aromas tot wyn moontlik verder evalueer word. Verdere navorsing is nodig om die moontlikheid van die bou en geldigheidsbepaling van sulke modelle te ondersoek.

Thesis (MScAgric (Viticulture and Oenology. Wine Biotechnology))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: The Vitis vinifera L. carotenoid cleavage dioxygenase 1 gene (VvCCD1) is a member of a structurally conserved gene family encoding enzymes that cleave multiple carotenoid substrates to form apocarotenoids. Carotenoid pigments are synthesised in the chloroplast where they are primarily involved in light harvesting and photo-protection during photosynthesis while apocarotenoids fulfill diverse roles that range from pollinator attractants to phytohormones. CCD1 cleaves carotenoids at specific double bond sites producing volatile apocarotenoids. These CCD1-derived apocarotenoids typically possess a fruity and floral aroma, thus making them desirable targets for metabolic engineering. CCD1 orthologues are highly homologous and have been isolated and characterised from a number of plant species, including Arabidopsis, tomato, rose, petunia, and grapevine. VvCCD1 is localised to the cytosol and has been shown in vitro to cleave zeaxanthin and lutein resulting in 3-hydroxy-β-ionone. Expression of VvCCD1 increases during berry ripening, peaking at véraison. Due to the impact that VvCCD1 potentially has on the flavour and aroma of grape berries and therefore wine, this study aimed to characterise the specific enzyme action as well as the biological role that this enzyme plays in grapevine. Expression of VvCCD1 in carotenoid-accumulating Escherichia coli strains demonstrated cleavage of β-carotene at the 9,10 (9’,10’) position forming β-ionone; and lycopene at the 5,6 (5’,6’) and 9,10 (9’,10’) position, forming 6-methyl-5-hepten-2-one and pseudoionone, respectively. A transgenic grapevine population with modified VvCCD1 expression was generated and genetically and metabolically characterised. The transgenic population consisted of lines in which VvCCD1 was either overexpressed or silenced. Expression analysis of stable transformants showed a 12-fold range of VvCCD1 expression relative to the wild-type. HPLC analysis of the photosynthetic pigment content of the transgenic population necessitated the development and optimisation of a method for the extraction of pigments, specifically from grapevine. A number of parameters were identified and optimised, resulting in a method that provides accurate quantification of photosynthetic pigments from grape berries and leaves. Absolute quantification of the following major photosynthetic pigments present in grapevine is now possible: chlorophyll a, chlorophyll b, lutein, -carotene, zeaxanthin, antheraxanthin, violaxanthin and neoxanthin. Data suggest that various levels of molecular control regulate carotenoid cleavage and apocarotenoid biosynthesis. The majority of lines stably transformed with a VvCCD1 overexpression cassette exhibit post-transcriptional gene silencing. Expression analysis in these lines demonstrated that, despite the additional contribution of transgene-derived VvCCD1 transcripts, the total VvCCD1 transcript levels were not significantly higher than in wild-type lines. In lines where transgenic manipulation of VvCCD1 expression was successful, subsequent analysis of carotenoids and apocarotenoids in leaf tissue showed no correlation between the measured metabolites and gene expression. The in planta action of VvCCD1 is presumably distinct from the observed in vitro activity due to the strict compartmentalisation required in photosynthetic leaf tissue preventing access of cytosolic VvCCD1 to the chloroplastic carotenoids. Future studies on reproductive organs (grape berries) from the transgenic lines generated in this study will be of great importance in further elucidation of the in planta function of VvCCD1. / AFRIKAANSE OPSOMMING: Die Vitis vinifera L. “carotenoid cleavage dioxygenase” 1 geen (VvCCD1) behoort aan ‘n geenfamilie wat struktureel gekonserveerd is en kodeer vir ensieme wat verskeie karotenoïed substrate afbreek om apokarotenoïede te vorm. Karotenoïed pigmente word in die chloroplaste gesintetiseer waar hulle primêr betrokke is by lig-insameling, sowel as beskerming tydens fotosintese, terwyl apokarotenoïede diverse funksies in die plant verrig wat strek van aantrekking van stuifmeelverspreiders tot phytohormone. CCD1 breek karotenoïede by spesifieke dubbelbindingsetels af om vlugtige apokarotenoïede te vorm. Die apokarotenoïede wat van CCD1 afkomstig is besit tipies vrugtige en blomagtige aromas wat hul gesogte teikens maak vir metaboliese manipulering. CCD1 ortoloë is hoogs homoloog en is al geїsoleer en gekarakteriseer vanuit ‘n verskeidenheid plantspesies wat Arabidopsis, tamatie, roos, petunia en wingerd insluit. VvCCD1 is in die sitosol gelokaliseer en dit is vantevore gewys dat dit beide zeaxanthin en lutein in vitro kan afbreek om 3-hidroksi-b-ionoon te vorm. Die uitdrukking van VvCCD1 vermeerder tydens korrel rypwording en bereik ‘n maksimum tydens véraison. Weens die potensieële invloed vanVvCCD1 op die geur en aroma van druiwe, en dus wyn, is hierdie studie gerig op die karakterisering van die spesifieke ensiematiese aksie, sowel as die biologiese rol van hierdie ensiem in wingerd. Uitdrukking van VvCCD1 in Escherichia coli rasse wat karotenoïede versamel het getoon dat β-karoteen by die 9,10 (9’,10’) posisie afgebreek word om β-ionoon te vorm, en likopeen by die 5,6 (5’,6’) en 9,10 (9’,10’) posisie om onderskeidelik 6-metiel-5-hepteen-2-oon en pseudo-ionoon te vorm. ‘n Transgeniese wingerd populasie is gegenereer met gewysigde VvCCD1 uitdrukking en is geneties en metabolies gekarakteriseer. Die transgeniese populasie het bestaan uit lyne waar VvCCD1 óf ooruitgedruk óf afgereguleer is. Uitdrukkings analise van die stabiele transformante het ‘n 12-voudige reeks van VvCCD1 uitdrukking getoon, relatief tot die wilde tipe. HPLC analise van die fotosintetiese-pigment inhoud van die transgeniese populasie het die ontwikkeling en optimisering van ‘n wingerd-spesifieke metode vir die ekstraksie van pigmente genoodsaak. ‘n Aantal parameters is geïdentifiseer en geoptimiseer, en het gelei tot ‘n metode wat akkurate kwantifisering van fotosintetiese pigmente in druiwe en wingerdblare kan lewer. Absolute kwantifisering van die volgende belangrike fotosintetiese pigmente aanwesig in wingerd is nou moontlik: chlorophyll a, chlorophyll b, lutein, -karoteen, zeaxantien, anteraxantien, violaxantien en neoxantien. Data dui aan dat verskeie vlakke van molekulêre beheer die afbreking van karoteen en die biosintese van apokarotenoïede reguleer. Die meerderheid van die lyne wat stabiel getransformeer is met ‘n VvCCD1 ooruitdrukkingskasset het na-transkripsioneleafregulering van die geen getoon. Uitdrukking analise van die lyne het gewys dat ten spyte van die addisionele transgeniese VvCCD1 transkripte, die totale VvCCD1 transkripvlakke nie beduidend hoër was as dié van die wilde-tipe lyne nie. In die lyne waar transgeniese manipulasie van VvCCD1 uitdrukking wel suksesvol was, het verdere analise van die karotenoïed en apokarotenoïed vlakke in blaarweefsel geen korrelasie getoon tussen die metaboliete en VvCCD1 uitdrukking nie. Die in planta aktiwiteit van VvCCD1 is vermoedelik anders as die in vitro aktiwiteit weens die streng kompartementalisering benodig in fotosintetiese blaarweefsel, wat verhoed dat die sitosoliese VvCCD1 toegang het tot die chloroplastiese karotenoïede. Toekomstige bestudering van die reproduktiewe organe (druiwe) van die transgeniese lyne wat in hierdie studie gegenereer is sal belangrik wees in die verdere verduideliking van die in planta funksie van VvCCD1.

Diabetes and cardiovascular disease are leading causes of death and disability in the United States. These chronic diseases are clinical sequelae of metabolic syndrome (MetS), a condition that affects approximately one-third (1/3) of American adults. Metabolic syndrome occurs in response to environmental and genetic influences, among them food intake, a sedentary lifestyle, BMI, advancing age, and exposure to persistent organic pollutants (POPs). POPs are known to cause endocrine disruption and PCBs cause oxidative stress, disrupt endothelial cell integrity, and promote atherosclerosis. Nutrition plays a significant role in the prevention and management of these chronic diseases and has been shown to modulate the toxicity of PCBs. Serum carotenoid (SC) concentrations are the best biomarker indicative of fruit and vegetable intake and an improved nutritional status. The purpose of this study was to investigate the relationship between serum carotenoid concentrations, serum concentrations of PCBs, and the probability of developing metabolic syndrome. The National Health and Nutrition Examination Survey (NHANES) is a program of the National Center for Health Statistics (NCHS), Centers for Disease Control and Prevention (CDC), that utilizes a cross-sectional sample survey design to collect, maintain, and disseminate the health and nutrition data of persons residing in the United States. Carotenoids and PCBs share similar biological pathways due in part to lipophilicity. Both concentrate to lipids in blood, are stored primarily in adipose tissue, and may competitively bind nuclear receptors. A statistical interaction was sought between the two variables for their combined effect on the probability of metabolic syndrome. An increase in probability was observed in the first exposure quartile for many PCBs, individually and pooled, suggestive of a low dose endocrine effect. Statistical modeling consistently showed strong decreasing trends in the probability of metabolic syndrome with higher concentrations of serum carotenoids in the 3rd and 4th PCB exposure quartiles. These data suggest a protective effect of serum carotenoids, and therefore of fruit and vegetable intake, despite higher serum levels of PCBs, in the probability of developing metabolic syndrome.