Thesis (PhD(Agric))--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Water scarcity is a key limiting factor for viticulture in dry regions. Traditionally drought sensitive varieties have the potential to grow in dry areas, however in most situations, through the use of rootstocks. Drought-tolerant rootstocks are expected to improve grapevine response to water deficit by improving the water uptake and transport and by reducing the water loss in leaves by root-to-shoot signalling. The mechanisms of rootstocks’ tolerance to drought are not yet fully understood. The main aim of this study was to improve the understanding of the rootstock/scion-cultivar interaction in the regulation of grapevine water use and leaf stomatal behaviour. Irrigated field vines without any water constraint were compared to rain-fed grapevines subjected to moderate water constraint. To better manage vine water status, reduce variability, and compare more rootstocks, greenhouse trials were also conducted where plants were well watered or subjected to severe water constraints. Pinotage grapevines (Vitis vinifera L.) grafted onto 110 Richter, 140 Ruggeri and 1103 Paulsen rootstocks were used for field experiments whereas Pinotage grapevines grafted onto 99 Richter, 110 Richter, 140 Ruggeri, 1103 Paulsen and Ramsey were used for greenhouse experiments. Our study suggested the influence of rootstocks on scion-cultivar water status and leaf stomatal size and density and gas exchange of the scion, implying an influence on water uptake and transport and a tight regulation of the stomatal conductance. Our data supported the hypothesis that the influence of rootstock in response to drought seemed to be higher under increasing water deficit up to a point where the plant water status is the main driver of the stomatal conductance and therefore photosynthesis regulation, considering the plant water status thresholds. In addition, the results suggested that stomatal development is affected by light, drought and possibly by rootstocks. Nevertheless, it is still not clear how the rootstock affects stomatal development and the link with scion-cultivar water use. It seems that the transpiration rate of leaves is more related to stomatal size than density. Thus one possible mechanism of Pinotage leaf adaptation to water constraints was structural during leaf growth, with a reduction in pore size to reduce plant water loss. The results showed that the rootstock is regulating the cultivar's stomatal size (anatomical changes during leaf growth) and functioning (stomatal regulation) through a complex signalling process. The effect of light on stomatal development is interesting in the context of canopy microclimate and canopy manipulation (choice of the vine architecture vs canopy size, in the context of climate change versus the possible increase in drought and water scarcity). The use of rootstocks is a long term investment which aims to provide resistance to soil pests and pathogens and to confer to the scion-cultivar drought and salt tolerance. The use of drought tolerant rootstocks is actually one of the most relevant practical solutions in dry terroir – units and in situations where water availability is limited. The understanding of the physiological and genetic mechanisms which govern scion-cultivar drought tolerance/behaviour induced by rootstocks is critical in terms of rootstocks choice in interaction with the scion-cultivar and is critical to assist breeding programs to create/select drought tolerant rootstocks.

The aim of this study is to determine the content of selected metals (As, Ba, Cd, Co, Cr, Cu, Mn, Mo, Ni, Se, Sn, Sr, Pb, V, Zn) in samples of leaves and grapes produced by ecologic agriculture and compare to results with leaves and grapes produced by integrated agriculture. The measured results used to assess the impact of agricultural technology on the quality of vines and grapes. The amount of elements was analysed using inductively coupled plasma mass spectrometry after previous digestion of sapmles using microwaves. Concentrations of elements in leaves and grapes from organic and integrated farming did not differ significantly only in the case of copper concentrations were measured higher concentrations in samples from organic farming. This difference is probably caused by different doses of authorized protective equipment based on copper in the organic and integrated farming.

L’activation des réponses immunitaires des plantes repose sur la reconnaissance de motifs moléculaires associés aux pathogènes (aussi appelés PAMP) par des récepteurs de l’immunité, également nommés PRR (pattern recognition receptors). La chitine, principal composant de la paroi des champignons, est un PAMP bien caractérisé qui induit des réponses de défense aussi bien chez les mammifères que chez les plantes.La première partie de cette étude met en évidence que deux chito-oligosaccharides, la chitine et le chitosan, agissent comme des PAMP chez la vigne (Vitis vinifera) puisqu’ils induisent des évènements précoces de signalisation, l’expression de gènes de défense et une résistance contre des agents pathogènes. Ces résultats suggèrent que des systèmes de perception existent chez la vigne. Une analyse phylogénétique a permis d’identifier trois récepteurs kinases à domaine LysM (LysM-RK ou LYK) chez V. vinifera (VvLYK1-1, -2, -3) appartenant au même clade que le récepteur à la chitine chez Arabidopsis et nommé AtCERK1 (Arabidopsis thaliana Chitin Elicitor Receptor Kinase 1). Leur analyse fonctionnelle a été réalisée par complémentation du mutant d’Arabidopsis Atcerk1, affecté dans la perception de la chitine. Nos résultats montrent que VvLYK1-1 et VvLYK1-2, mais pas VvLYK1-3, complémentent fonctionnellement le mutant Atcerk1 en restaurant l’activation des MAPK (Mitogen-Activated Protein Kinases) et l’expression de gènes de défense induits par les chito-oligosaccharides. De plus, l’expression de VvLYK1-1 chez Atcerk1 restaure la résistance basale à l’agent de l’oïdium de la vigne (Erysiphe necator).La seconde partie du projet s’est focalisée sur les éliciteurs oligosaccharidiques de type « damage-associated molecular patterns (DAMP) ». Ces molécules endogènes peuvent provenir de la dégradation de la paroi lors d’une attaque et sont capables d’activer les réponses immunitaires de la plante. Les DAMP les mieux caractérisés actuellement sont les oligogalacturonates (OG), des fragments de pectine qui induisent des réponses immunitaires chez de nombreuses espèces végétales dont l’activation de MAPK, la production d’H2O2, l’expression de gènes de défense et le dépôt de callose. Nous avons montré dans cette étude que les xyloglucanes (Xh), des fragments d’hémicellulose pariétale purifiés, induisaient l’activation de MAPK et l’expression de gènes de défense chez la vigne et Arabidopsis, afin d’induire une résistance contre le champignon nécrotrophe Botrytis cinerea. Les Xh induisent également la production de resvératrol, une phytoalexine majoritaire chez la vigne, et un dépôt de callose chez Arabidopsis. Par une approche génétique, nous avons identifié certains composants de la signalisation induite par les Xh chez Arabidopsis. L’utilisation de mutants suggère que la résistance induite par les Xh contre B. cinerea est dépendante des voies de la camalexine, de l’acide salicylique, de l’acide jasmonique et de l’éthylène chez Arabidopsis. De manière globale, nos résultats mettent en lumière que les xyloglucanes peuvent être considérés comme de nouveaux éliciteurs de l’immunité chez la vigne et Arabidopsis. / Activation of the plant immune responses requires recognition of common pathogen-associated molecular pattern (PAMP) by their cognate pattern recognition receptors (PRR). Chitin, a major component of fungal cell walls, is a well-known PAMP that triggers defense responses in several mammal and plant species.In the first part of this study, we show that two chitooligosaccharides, chitin and chitosan, act as PAMPs in grapevine (Vitis vinifera) as they elicit immune signaling events, defense gene expression, and resistance against pathogens. These two PAMPs are active in grapevine suggesting that at least one perception system exists. Phylogenetic analysis clearly distinguished three V. vinifera LysM Receptor Kinases (VvLYK1-1, -2, -3) located in the same clade as the Arabidopsis Chitin Elicitor Receptor Kinase 1 (AtCERK1), which mediates chitin-induced immune responses. Their functional characterization was achieved by complementation assays in the Atcerk1 mutant, impaired in chitin perception. Our results provide evidence that VvLYK1-1 and VvLYK1-2, but not VvLYK1-3, functionally complement the loss of AtCERK1 function by restoring chitooligosaccharide-induced MAPK activation and immune gene expression. Moreover, expression of VvLYK1-1 in Atcerk1 restored penetration resistance to the non-adapted grapevine powdery mildew (Erysiphe necator).The second part of this study focused on damaged-associated molecular patterns (DAMP), endogenous molecules that can be released from the plant cell wall during an attack and activate the plant innate immunity. Until now, the best characterized DAMPs are oligogalacturonides (OG) coming from pectin fragments that induce innate immune responses in various plant species, including MAPK activation, H2O2 production, defense gene expression and callose deposition. In this study, we showed that purified xyloglucans (Xh), derived from the plant cell wall hemicellulose, elicit MAPK activation and immune gene expression in grapevine (V. vinifera) and Arabidopsis to trigger induced resistance against the necrotrophic fungus Botrytis cinerea. Xh also elicit the production of resveratrol, the main grapevine phytoalexin, and callose deposition in Arabidopsis. Using a genetic approach, we identified some signaling components of Xh-induced immunity. The use of Arabidopsis mutants suggests that Xh-induced resistance against B. cinerea is dependent on the camalexin, salicylate, jasmonate and ethylene pathways. Taken together, our data highlight that Xh can be considered as new elicitors of grapevine and Arabidopsis immunity.