Worldwide production of potatoes is in excess of 350 million metric tonnes per annum, of which 60% is intended for human consumption. Since the period of tuber dormancy before tuber buds sprout is usually shorter than the optimal market storage period, control of sprouting is essential. To prolong dormancy, tubers are either stored at low temperatures and/or are treated with chemical sprout inhibitors, the use of which subject to increasing scrutiny in the European Union due to their impact upon the environment. Cytokinins, a group of plant growth regulators, are known to play a central role in tuber bud sprouting and tuber initiation from stolon tips in Solanum tuberosum L. although it is unclear when and how cytokinins act to regulate dormancy. The interconversion of cytokinins is incompletely understood. Enzymes identified to date have higher affinities for aminopurines than their cytokinin equivalents. A novel cytokinin binding protein Solanum tuberosum Cytokinin Phosphorylase 1 (StCKP1), has been identified in tuberising stolon tips which shares regions of homology with members of the nucleosidase and phosphoribosyl transferase family. The composition of cytokinin N9 conjugates in tuber bud and surrounding tissue is known to change on transition from a dormant state, with an increase in base and riboside types observed. Analysis of transcripts indicates an increase in abundance of StCKP1 on tuberisation of stolon tips, and high abundance in the periderm of dormant tubers. Analysis of protein abundance by immunoblotting echoes this finding and indicates StCKP1 begins to accumulate in stolon tips shortly before tuberisation, matching binding activity. Transgenic analysis of the cytological reporter gene uidA under the control of two identified promoter regions indicates StCKP1 is expressed predominantly in tuber tissue. Analysis of StCKP1 activity by HPLC and LC-MS-MS shows that StCKP1 catalyses the interconversion of free base and riboside. KMs determined for cytokinin and aminopurine substrates indicate that StCKP1 has a higher affinity for cytokinin substrates and, of these cytokinins, displays a higher affinity for the free base catalysing ribosylation of the N9 to form the corresponding riboside. Desiree cultivars over-expressing StCKP1 under the CaMV 35S promoter exhibited an increased rate of tuberisation of stolon tips and an increase in the length of the dormant period following lifting. Over-expression of StCKP1 was found in particular to increase the chill sensitive period of dormancy, confirming results of StCKP1 knock-down by RNAi. Transcript abundance of StCKP1 at tuberisation in other cultivars including King Edward and Maris Peer was found to correlate with the dormancy characteristics prescribed by the European Cultivated Potato Database and the British potato variety database.

The use of various types of gel electrophoresis for characterization and identifying plant varieties is well estabilished. This diploma work deals with the possibility characterize potato cultivar (Solanum tuberosum L.) by the help of electrophoretic technique SDS {--} PAGE tuber protein profiles. Soluble potato tuber proteins can be classified according to their molecular weight (kDa) into three groups: First, the major tuber protein patatin (37 {--} 44 kDa); second, a group constituted of protease inhibitors (6 {--} 24 kDa); and third group containing all other proteins. First two group manifest genetic variability, which is necessary evalute for its potential using of potato cultivar characterization.

La pomme de terre produit des glycoalcaloïdes comme la plupart des Solanacées. Deux composés, en particulier, l’α-solanine et l’α-chaconine, sont produits en plus grande quantité lorsque le tubercule est exposé à la lumière et subit des dégradations. Ces molécules sont toxiques et peuvent représenter un danger pour le consommateur et des nuisances pour l’environnement. Dans le cadre de cette thèse, les teneurs en α-solanine et α-chaconine dans la chaire de pomme de terre verdie, en germination ou en décomposition retrouvées sur les marchés d’Abidjan (Côte d'Ivoire) ont été analysées. Les résultats ont montré que la chaire de ces pommes de terre contient des quantités élevées de ces deux composés, dépassant 2 à 5 fois la limite recommandée. Pour des raisons de sécurité sanitaire, ces pommes de terre doivent être proscrites de l’alimentation humaine. En revanche, la teneur élevée en glycoalcaloïdes dans ces pommes de terre représente une source de solanidine, un précurseur pour la synthèse d'hormones et de composés pharmacologiquement actifs, qui mérite d’être exploitée. Dans cet objectif, nous avons développé une méthode chimio-enzymatique simple, comprenant un traitement acide partiel suivi d’une hydrolyse enzymatique par la β-glycosidase de la blatte Periplaneta americana dont le gène a été isolé à partir d’une librairie génomique de cDNA afin de détoxifier ces composés et produire la solanidine. / Potato produce glycoalkaloids as most plants of Solanaceae family. The principal glycoalkaloids, α-chaconine and α-solanine are produced in greater quantities when potato tubers are exposed to light and are subject to deteriorations. These compounds are toxic and can represent a real danger for the consumer and the environment where they are discharged during their degradation. In this work, the estimation of glycoalkaloids in the flesh of different types of decayed potatoes usually found in different market places of Abidjan (Ivory Coast) was evaluated. The results showed that turned green and also sprouting or rotting potato flesh contain high amounts of toxic solanine and chaconine, exceeding by 2 to 5-fold the recommended limit. For safety consideration, these decayed potatoes should be systematically set aside. The accumulation of α-chaconine and solanine in potatoes can be seen as an attractive source of solanidine that is an important precursor for hormone synthesis and some pharmacologically active compounds. To this end, we proposed herein a simple chemo-enzymatic protocol comprising a partial acidic hydrolysis followed by an enzymatic treatment with the β-glycosidase from Periplaneta americana whose gene was isolated from a cDNA genomic library in order to detoxify these compounds and produce solanidine.