Fusarium wilt of banana has a long and devastating history in many of the world’s banana producing countries. The most pronounced damage caused by Fusarium oxysporum f.sp. cubense (Foc), the Fusarium wilt pathogen, occurred during the 20th century in Central America, where tens of thousands of virgin forests were lost to further banana production. No control strategy is effective against Fusarium wilt other than replacement of susceptible by resistant varieties. It is, therefore, important to develop or identify resistant replacements that would not only be able to resist the pathogen, but also be acceptable to consumers. Resistance in wild banana varieties has been identified, and hybrids have been developed by breeding programmes with good resistance to Fusarium wilt. These varieties, unfortunately, appear not to be acceptable replacements for Cavendish bananas, the sweet desert banana variety that serves as the primary export banana and constitutes almost 40% of all bananas planted in the world today. A field selection, GCTCV-218, now proved to be the Cavendish plant with the most resistance to Foc ‘tropical’ race 4 (VCG 0121) has saved the Cavendish-based banana industry in Taiwan from devastation. In this thesis, GCTCV-218 has been evaluated against Foc ‘subtropical’ race 4 (VCG 0120), the primary variant of the pathogen in subtropical banana-producing countries such as South Africa, Australia and the Canary Islands. Defence-associated genes that are differentially expressed and that were up-regulated early in the defence response against the pathogen were isolated and identified. Greenhouse and field trials conducted at the research facilities of the Forestry and Agricultural Biotechnology Institute, University of Pretoria and in Kiepersol, South Africa, respectively, showed that GCTCV-218 had a significantly higher level of disease tolerance against Foc ‘subtropical’ race 4 (VCG 0120) when compared to the commercially grown Williams cultivar. Phenolic assays revealed that total phenolics and cell-wall bound phenolics were expressed at higher levels in GCTCV-218 after pathogen attack and seemed to play an important role in the tolerance of GCTCV-218. It was, therefore, proposed that GCTCV-218 could be considered a replacement for other Cavendish banana varieties planted in South Africa. The genetic basis of defence mechanisms in banana to Foc is unknown. In this investigation, Suppression Subtractive Hybridisation (SSH) was used to construct a cDNA library, containing banana genes that were up-regulated early (3&6 hours after infection), in the GCTCV-218/Foc interaction. The efficiency of the procedure was confirmed by PCR amplification of a known defence gene (endochitinase) present in the subtracted tester material, as well as analysing the reduction of a known housekeeping gene, actin, in the subtracted material compared to unsubtracted material. Southern blot data further provided confidence in the subtraction process. A cDNA library containing 736 gene fragments was constructed and then subjected to a screening procedure to remove false positives that escaped the subtraction process. The screening of a banana cDNA library for defence-related genes involved the development of a high-throughput cDNA microarray technique. This novel technique removed all false positives, such as housekeeping genes that escaped the subtraction as well as clones representing rDNAs. Seventy-nine genes differentially expressed in GCTCV-218 and not in Williams were selected, sequenced and subjected to BLASTX, BLASTN and DBest searches. Of these, several gene fragments showed homology to defence-associated genes, and 20 unique genes fragments were identified. These include two different peroxidases, response regulator 6, catalase 2, metallothionein, pectin acetylesterase (PAE), two different unknown proteins, salt stress, trypsin inhibitor, unspecific monooxygenase cytochrome P450, Bowman Birk proteinase inhibitor, root control, xylanase inhibitor, inhibitor CII, hypothetical protein, putative senescence-associated protein, pathogenesis-related protein 1 (PR1) and ribosomal protein S3a. The significance of the defence reaction to Fusarium wilt diseases in agricultural crops depends on the tempo of plant response. When a host plant is able to respond early to pathogen invasion the pathogen is successfully contained, preventing further spread throughout the plant. The expression of genes with antimicrobial activity, such as endochitinase, suggests an induced biochemical defence response against Foc. The expression of PAE and PR1 results in the deposition of lignin and callose production for cell wall strengthening. Four defence associated genes (catalase 2, pectin acetyl esterase (PAE), PR-1 and endochitinase) were selected for expression profile analysis using Real-time reverse transcriptase PCR, with TaqMan® and Light Cycler technology. All four genes were shown to be differentially expressed in GCTCV-218 at 3 and 6 hrs after infection, confirming SSH results. PR-1 and PAE were induced very early (3 hrs after infection) in the GCTCV-218, while PR3 and catalase 2 followed with a significant induction at 6 hrs after infection. This study concludes that GCTCV-218 is able to respond rapidly in response to Foc infection by activating both a biochemical and structural defence mechanism. / Thesis (PhD (Plant Pathology))--University of Pretoria, 2006. / Microbiology and Plant Pathology / unrestricted

Nos travaux ont permis d'étudier l'effet combiné de la maturation, du mûrissement et des périodes climatiques, sur le profil des composés bioactifs de la banane Musa Cavendish, issus des French West Indies (FWI). La teneur en polyphénols des bananes 400 Degrés Jour (dj) est restée inchangée, tandis que les bananes 600 et 900 dj ont présenté une augmentation significative de leurs teneurs en polyphénols pendant la maturation (p<O,OS). En outre, les résultats ont également montré que, bien que la dopamine ait été déterminée comme étant le polyphénol le plus élevé dans la peau de bananes pendant le stade verte de développement et la maturation, sa cinétique différait du profil observé pour celle des polyphénols totaux. Les facteurs post-récolte ont été optimisés. L'ensemble de nos résultats ont montré que la sélection d'une matrice de choix tenant compte de la maturation, du mûrissement et du climat permettait d'atteindre un statut optimal de Musa Cavendish pour des concentrations optimales en composés identifiés pertinents pour la santé humaine. C'est effectivement le cas des concentrations de dopamine, médiateur vasoactif, dépendantes des stades physiologiques, du mûrissement et climat. Nos travaux ont ainsi démontré que le totum de banane présentait un effet contracturant dose dépendant sur l'aorte moins puissant que celui enregistré pour les composés de référence à concentrations plus faibles mais toutefois significatifs. Dans les mêmes conditions, il a été démontré que la prazosine modifie l'effet contracturant de la banane en l'inhibant de manière compétitive. Le récepteur alpha-l adrénergique vasculaire joue un rôle dans les effets vasoconstricteurs de la banane. Les phénomènes ulcéreux étudiés à travers l'inhibition ou non de la pousse de Helicobacter pylori après addition ou pas d'extraits polaires et apolaires de bananes ont indiqué le rôle non-inhibiteur de Musa cavendish sur la croissance de Helicobacter pylori. / The combined influence of maturation, ripening and climate periods on French West Indies (FWI) bananas, Musa Cavendish, bioactive compounds profile was studied. The polyphenol content of the 400 dd bananas remained unchanged, while the 600 and 900 dd bananas exhibited a significant polyphenol increase during ripening (p<O.OS). Furthermore, the results showed that although dopamine was deterrnined as the highest polyphenol in the banana peels during the green developmental stage and ripening, its kinetic differed from the total polyphenol profile. Together ours results showed that this matrix of choice (maturation, ripening and climate may allow selection of Musa Cavendish status for optimal concentration in identified compounds with human health relevance. Indeed, this is the case of dopamine, a vasoactive mediator, which is dependent on physiological stages, ripening and climate. Thus, our work has demonstrated that the banana totum showed a dose-dependent contracture effect on aortic which was less powerful than the one recorded for reference compounds at lower concentrations. Nevertheless it was significant. In the same conditions, it was demonstrated that prazosin modifies the effect of banana contracture inhibiting it competitively. Receptor alpha-I adrenergic vascular plays a role in the vasoconstrictor effects of bananas. The study of ulcers through inhibition! non inhibition of Helicobacter pylori growth after addition of polar and apolar banana extracts indicated the role of Musa cavendish as a non-inhibitor of Heiicobacter pylori growth