African leafy vegetables have been shown and suggested to have potential to contribute to human diets and alleviate malnutrition; however, their levels of utilisation are currently low especially in South Africa. This is because there is limited access to these crops due to low availability in the market. Limited access is attributed, in part, to the lack of commercialisation as a result of limited agronomic information describing optimum management options for these leafy vegetables. Availability of such information would contribute to successful commercialisation of these crops. The primary objective of this study was to establish optimum agronomic management factors for Amaranthus cruentus, Corchorus olitorius, Vigna unguiculata and Brassica juncea for irrigated commercial production in South Africa. Seeds of Amaranthus cruentus, Corchorus olitorius were obtained from the Agricultural Research Council seed bank; Vigna unguiculata were obtained from Hydrotech and Brassica juncea seeds were obtained from Stark Ayres. The project consisted of three field studies whose overall objective was to evaluate growth and yield responses of the selected African leafy vegetables to agronomic factors under irrigated commercial production. These field studies comprised of two single factors; summer trials (planting density and nitrogen on three selected crops) and a combined winter trial (nitrogen, irrigation, plant density and planting date on a winter crop). Chapter three (3) investigated the effect of plant density on growth, physiology and yield responses of Amaranthus cruentus, Corchorus olitorius and Vigna unguiculata to three plant densities under drip irrigated commercial production. The plant density levels of 100 000, 66 666 and 50 000 plants/ha were used in the 2011/12 and 2012/13 summer seasons. Parameters measured included chlorophyll content index (CCI), chlorophyll fluorescence (CF), stomatal conductance (SC), leaf number, leaf area index (LAI) and biomass. Amaranthus cruentus and Corchorus olitorius showed better leaf quality at lower plant density of 50 000 plants ha-1 than at 66 666 plants ha-1 and 100 000 plants ha-1. These results are based on bigger leaves expressed as leaf area index (LAI), better colour expressed as chlorophyll (CCI) and higher biomass per plant observed in these crops at 50 000 plants ha-1 in comparison to 66 666 plants ha-1 and 100 000 plants ha-1. In Vigna unguiculata there were no responses observed in LAI and CCI. In Amaranthus cruentus, Corchorus olitorius and Vigna unguiculata fresh and dry mass yield of leaves were higher at 100 000 plants ha-1 compared to other treatments. In A. cruentus and C. olitorius, higher leaf quality parameters (CCI, plant height, leaf number, biomass per plant and LAI) indicated that these crops can perform better at lower densities of 50 000 than at 66 666 plants ha-1 and 100 000 plants ha-1 Therefore, using 50 000 plants ha- 1 is suitable for commercial production of A. cruentus and C. olitorius. In Vigna unguiculata, a plant density of 100 000 plants ha-1 produced the highest fresh and dry mass per unit area without compromising quality in terms of the leaf size (LAI) and colour (CCI). Therefore 100 000 plants ha-1 is a density recommended for commercial production in V. unguiculata.Chapter four (4) was conducted to investigate growth, physiology and yield responses of A. cruentus, C. olitorius and V unguiculata to nitrogen application under drip irrigated commercial production. Three nitrogen treatments levels were used viz. 0, 44 and 88 kg N ha- 1 in 2011/12 season and four nitrogen treatments levels viz. 0, 50, 100 and 125 kg N ha-1 were used in 2012/13 summer season. The nitrogen levels selected for each season were based on recommendations for Amaranthaceae species, Swiss chard (Beta vulgaris L.var cicla) derived from soil analysis of the trial (field) site. Parameters measured included chlorophyll content index (CCI), chlorophyll fluorescence (CF), stomatal conductance (SC), leaf number, leaf area index (LAI) and biomass. Results showed that application of nitrogen at 44 kg N ha- 1 in 2011/12 summer season and 100 kg N ha-1 in 2012/13 summer season improved LAI, CCI, biomass per plants and yield in A. cruentus. A similar trend was observed in C. olitorius except that 44 kg N ha-1 improved stem fresh yield. Further increase in nitrogen fertiliser above 44 kg N ha-1 during the 2011/12 season and above 100 kg N ha-1 in 2012/13 summer season reduced leaf quality and yield in both crops. In V. unguiculata, nitrogen application showed a slight increase in yield values from 0 to 44 kg N ha-1 followed by decrease at 88 kg N ha-1 in 2011/12 summer season; however, this increase in yield was not significant. During the 2012/13 summer season, yield in terms of fresh weight was significantly (P<.001) reduced by applying nitrogen at various levels. However, leaf dry matter content increased significantly (P<.001) with increase in nitrogen from 0 kg up to 100 kg N ha-1, then remained unchanged at 125 kg N ha-1. Therefore, the current study recommends that C. olitorius and A. cruentus could be commercialised at 44 kg N ha-1 and 100 kg N ha-1 which were lower nitrogen application rates than those recommended for Amaranthaceae species. In V. unguiculata, 50 kg N ha-1 improved leaf number; however, this did not translate to any fresh yield advantage, implying that the optimum rate for nitrogen application might be lower than 50 kg N ha-1. Therefore, nitrogen rates less than the ones used in the current study are recommended for V. unguiculata. Chapter five (5) was conducted in winter and it was necessitated by observations made primarily in the previous studies which focused on the effects of single factors such as plant density, planting date and nitrogen deficits. Therefore, there was a need to address interactions between irrigation, nitrogen, spacing and planting date. The objective of this study was to evaluate growth, physiology and yield responses of Brassica juncea to different agronomic and management factors in the 2012 and 2013 seasons. The treatments were as follows: two planting dates in main plot (1 June and 18 July, 2012); two irrigation frequency in sub main plot (once and three times a week); three nitrogen levels (0, 50, 100 kg N ha-1) and three plant densities (133 333, 80 000, 50 000 plants ha-1) as subplots. Parameters measured included chlorophyll content index (CCI), chlorophyll fluorescence (CF), stomatal conductance (SC), leaf number, leaf area index (LAI) and biomass. Results from this study showed a significant interaction effect on plant height, LAI, CCI and CF. Crops irrigated thrice or once a week with 50 kg N ha-1 combined with 50 000 plants ha-1 produced tall plants and bigger leaves (LAI) in the early planting date (1 June) compared to other combinations. Irrigating three times a week combined with nitrogen application at 100 or 50 kg N ha-1 improved CF for late planting date (18 July) in comparison to other combinations. Irrigating once a week combined with nitrogen application at 100 kg N ha-1 increased CCI. There was no significant interaction effect on yield. Application of nitrogen at 50 and 100 kg N ha-1 significantly (P>0.05) increased yield in early and late planting dates compared to the control (0 kg N ha-1), in 2012 and 2013 winter season. Irrigating three times a week led to a significant (P<0.05) increase in yield in the late planting date (18th July) and early planting date (1st June) in 2013 season. Higher plant density of 133 333 plants ha-1 resulted in significantly (P<0.05) higher yield in terms of fresh mass and leaf number in the late planting date 18 July in 2012 and 2013 seasons. However, leaf quality parameters such as leaf size and colour was compromised at 133 333 plants ha-1 relative to 50 000 plants ha-1. Therefore, farmers are recommended to plant early, apply 50 kg N ha-1, irrigate thrice a week and utilise a spacing of 50 000 plants ha-1. The current study indicates that growth and yield of traditional leaf vegetables can be optimised through improved agronomic practise. / Agriculture and Life Sciences / D. Litt. et. Phil. (Agriculture)

Virtuellement toutes les plantes hébergent des champignons et des bactéries endosymbiontes (endophytes). Ces microorganismes façonnent le développement de leur hôte et peuvent inhiber des phytopathogènes. Au niveau moléculaire, les interactions plante-endophyte sont médiées par des molécules secrétées y compris des protéines et métabolites secondaires. Au cours des dernières années, la recherche d’endophytes a augmenté chez nombreux plantes, cependant chez les Ericaceae les endophytes ne sont pas bien connus. Alors, on s’est mis à investiguer les endophytes racinaires de la canneberge, une plante membre d’Ericaceae native de l’Amérique du Nord. On a échantillonné quatre plants provenant d’une ferme commerciale organique. Au total, 30 souches fongiques et 25 bactériens ont été isolés. Les bactéries Pseudomonas sp. EB212, Bacillus sp. EB213 et EB214; et les champignons Hyaloscypha sp. EC200, Pezicula sp. EC205 et Phialocephala sp. EC208 ont supprimé la croissance de cinq pathogènes de la canneberge, incluant Godronia cassandrae, un champignon causant la pourriture des fruits de la canneberge au Québec. EB213 a été capable de promouvoir légèrement la croissance de plantules de la canneberge. En performant des techniques microscopiques, on a constaté l’habileté de EC200, EC205 et EC208 à coloniser internement les racines des plantules de la canneberge. De plus, les génomes de ces champignons ont été séquencés, assemblés et annotés. Les analyses génomiques se sont concentrées sur les protéines secrétées et les groupes des gènes impliqués dans la biosynthèse (GGB). On a trouvé un large répertoire de gènes codant pour des enzymes qui métabolisent les carbohydrates et d’autres codant pour des protéases. Les deux groupes d’enzymes seraient utiles à dégrader de la matière organique pour libérer des nutriments. Aussi bien, ces enzymes pourraient faciliter la colonisation des racines de la plante hôte. De plus, on a prédit des nombreuses protéines effectrices qui assisteraient les endophytes à éviter l’activation du système immunitaire des plants. A noter que parmi les GGB inférés dans les génomes de EC200, EC205 et EC208, environ 90% ne sont pas caractérisés. Finalement, on a performé des analyses transcriptomiques pour élucider la réponse de EC200, EC205 et EC208 envers la présence de leur hôte, simulée par l’addition d’un extrait de canneberge au milieu de culture. Les conclusions majeures sont que les racines des plantes de la canneberge qui ont été échantillonnées sont dominées par des microorganismes avec l’habileté d’inhiber des phytopathogènes ; et que les génomes de EC200, EC205 et EC208 codent pour un grand répertoire de protéines qui pourraient être liées aux interactions plante-endophyte. / Virtually all plants host fungal and bacterial endosymbionts (endophytes). These microbes shape plant development and may inhibit phytopathogens. At the molecular level, plant-endophyte interactions are mediated by secreted compounds, including proteins and secondary metabolites. While endophytes are increasingly studied in diverse plants, little is known about their presence in Ericaceae. Therefore, we set out to investigate the root endophytes of cranberry, an ericacean member native to North America. We sampled endophytes from four plants grown on an organic farm. In total, 30 fungal and 25 bacterial strains were isolated and identified. A subset of these, notably Pseudomonas sp. EB212, Bacillus sp. EB213 and EB214; and fungi Hyaloscypha sp. EC200, Pezicula sp. EC205, and Phialocephala sp. EC208, were tested for their ability to suppress phytopathogens. Altogether, they inhibited five cranberry pathogens, including Godronia cassandrae, an important cranberry fruit-rot agent in Quebec. EB213 was the only endophyte that increased the biomass of cranberry seedlings. Using microscopy techniques, we confirmed the ability of EC200, EC205, and EC208 to colonize cranberry roots internally. The genomes of these fungi were sequenced, assembled and annotated. Genomic analyses focused on secreted proteins and biosynthetic gene clusters (BGCs). We found an extensive repertoire of carbohydrate-active enzymes and proteases that could assist in recycling organic nutrients, rendering them accessible to plants; these enzymes may also facilitate root colonization. In addition, effector proteins were predicted; these molecules may assist endophytes to escape the plant immune system and favour colonization. We inferred 139 biosynthetic gene clusters (BGCs) across the three examined fungi. Remarkably, the product of around 90% of BGCs are unknown. Finally, transcriptomic analyses were performed to determine how EC200, EC205 and EC208 respond to the presence of cranberry, simulated by the addition of cranberry extract in the culture medium. The two major conclusions of this work are that the roots of the sampled cranberry plants are dominated by endophytes with biocontrol abilities, and that EC200, EC205 and EC208 encode a broad repertoire of proteins that could be involved in plant-endophyte interactions.

Tomate (S. lycopersicum L.) und Ackerbohne (P. vulgaris L.) sind ein wichtiges landwirtschaftliches Erzeugnis, das weltweit angebaut wird. Ertragseinbußen bei diesen Kulturen werden unter anderem durch die Anfälligkeit für das Pepino mosaic virus und Rhizoctonia solani verursacht. In der vorliegenden Studie wurde daher das Potential von Endophyten und kommerziell formulierten produkten - RhizoVital 42® fl., FZB24® WG, T-Gro, Prestop®, und Cérès sowohl in vitro als auch in vivo bei Tomaten und zur Unterdrückung von durch PepMV oder R. solani ausgelösten Erkrankungen oder zur Förderung des Pflanzenwachstums ermittelt. Bei den in-vivo-Studien die individuelle Behandlung von PepMV-infizierten Tomatenpflanzen mit Prestop® und T-Gro führte zu einer Steigerung der Pflanzenbiomasse bzw. der Pflanzenhöhe im Vergleich zu gesunden, unbehandelten Kontrollpflanzen. Die ANOVA-Analyse zeigte eine geringere PepMV-Akkumulation bei PepMV-infizierten Pflanzen, die dreimal mit Prestop® behandelt wurden, im Vergleich zu PepMV-infizierten und unbehandelten Kontrollpflanzen. Die Ko-Behandlung mit RhizoVital 42® fl. und T-Gro war bei Ackerbohnen am erfolgreichsten bei der Unterdrückung des Ausmaßes der Rhizoctonia-Wurzelfäule und bei der Verbesserung des Pflanzenwachstums und der Ertragsparameter im Vergleich zu den mit R. solani infizierten und unbehandelten Kontrollpflanzen. Eine Co-Inokulation mit T-Gro und RhizoVital 42® fl. steigerte den Tomatenfruchtertrag der R.solani-infizierten Tomatenpflanzen im Vergleich zu den R. solani infizierten unbehandelten Kontrollpflanzen. Unabhängig davon, ob die BCAs einzeln oder in Kombination angewendet wurden, kam es nach jeder Behandlung zu einem verbesserten Pflanzenwachstum und einer starken antagonistischen Aktivität gegen R. solani. Diese Ergebnisse unterstreichen das Potenzial des Einsatzes der Mikroorganismen und ihres Konsortiums als Biodünger zur Verbesserung des Pflanzenwachstums und der Kontrolle von RRR bei Bohnen- und Tomatenpflanzen. / Tomato (S. lycopersicum L.) and common bean (P. vulgaris L.) are valuable agricultural commodities worldwide. Yield losses in these crops are caused by, susceptibility to Pepino mosaic virus and R. solani. This study sought to evaluate the efficacy of endophytes and commercially formulated products- RhizoVital 42® fl., FZB24® WG, T-Gro, Prestop® and Cérès, both in vitro and in vivo, for their potential to suppress PepMV infection and Rhizoctonia root rot (RRR) and promote plant growth in tomato and beans. Under in vivo studies, treatment of PepMV-infected tomato plants with Prestop® and T-Gro enhanced plant biomass and height, respectively compared to healthy untreated control plants. ANOVA analysis showed lower levels of PepMV accumulation in PepMV-infected plants treated with Prestop® thrice compared to other bioagents. Co-inoculation with RhizoVital 42® fl. and T-Gro was the most successful in suppressing RRR severity in beans. Treatment with T-Gro significantly recorded the highest number of pods and total number of seeds per plant. In tomato, application of RhizoVital 42® fl. exhibited the strongest suppressive effect against RRR. Co-inoculation with T-Gro and RhizoVital 42® fl. significantly enhanced the total yield of fruits in R. solani-infected plants compared to R. solani-infected untreated controls. Irrespective of whether the BCAs were applied singly or in combination, there was improved growth and potent antagonistic activity against R. solani which highlights the potential of using bioagents and their consortium as biofertilizers for the improvement of plant growth and control of RRR in common beans and tomatoes.

Untersuchungen wurden durchgeführt, um die Wirkung von B. subtilis und deren Metaboliten auf den Meloidogyne- und Pratylenchus-Befall und ihre Vermehrung festzustellen sowie um die möglichen Wirkmechanismen zu studieren. Substratbehandlungen mit B. subtilis FZB 24® führten zu einem höherenMeloidogyne-Befall und einer verstärkten Nematodenvermehrung. Trotz verstärktem Befall wurde das Pflanzenwachstum verbessert (induzierte Toleranz). Weiterhin wurden durch "antibiotikafreie" Kulturfiltrate (KF) aus der bakterielen Übergangs- und stationären Phase ähnliche Wirkungen erzielt. Außerdem wurde eine systemische Wirkung von B. subtilis auf den Meloidogyne-Befall an Tomate nachgewiesen. B. subtilis bzw.die KF-behandelte Testpflanzen zeigten stärkere Anlockwirkung auf Meloidogyne-Larven (M. incognita, M. arenaria) als unbehandelte. KF (50, 10, 1%) von B. subtilis zeigten keine nematizide Wirkung auf die Meloidogyne-Larven. KNO3 als Trägersubstanz für das Bakterienpräparat besaß ähnliche Wirkungen auf den Meloidogyne-Befall und die Nematodenvermehrung.Ebenfalls wurde das Pflanzenwachstum durch KNO3-Zufuhr gefördert. Der Einsatz des nematodenfangenden Pilzes Arthrobotrys superba reduzierte den Meloidogyne-Befall (30% Gallenreduktion). Hingegen wurde durch die kombinierte Anwendung von A.superba und B. subtilis FZB 24® der Bekämpfungserfolg von A. superba reduziert. Die exogene Applikation von Phytohormonen bzw. Präkursoren zeigte keine Wirkung auf das Wachstum der Testpflanzen. Die Vermehrung von Meloidogyne wurde durch IAA und die Kombination von IAA und Kinetin gefördert. In den getesteten Konzentrationen der Phytohormone wurde keine Wirkung auf die Mortalität der Wurzelgallenälchen- (Meloidogyne-) Larven beobachtet. Der Gehalt von Enzymen (Chitinase, Glucanase und Peroxidase) aus dem Sproß behandelter Tomatenpflanzen wurde bestimmt. B. subtilis-Isolate (FZB 24® , S18) reduzierten die Population von Wurzelläsionsnematoden, Pratylenchus penetrans (nicht signifikant) [9 bzw. 15-20% pro Wurzelsystem bzw. g Wurzel]. Das Pflanzenwachstum wurde an befallenen Pflanzen durch beide Isolate verbessert (induzierte Resistenz). Es wurden keine Unterschiede zwischen den Bakterien-Isolaten festgestellt. KNO3 führte ebenfalls zu einer Verminderung der Nematodenpopulation. Die Ergebnisse werden hinsichtlich möglicher Wirkmechanismen des RhizobakteriumsB. subtilis und des nematodenfangendes PilzesA. superba zur Regulierung der Nematodenpopulation bei Tomate diskutiert. / Investigations were made to know about the effects of Bacillus subtilis and its metabolites on the infestation of tomato plants with root-knot and lesion nematodes. Further more, experiments were carried out to clear up the mode of actions ofB. subtilis and its culture filtrates on infestation of tomato seedlings and reproduction of root-knot nematodes. Substrate applications ofB. subtilis FZB 24® leaded to an increasement of infestations intensity and reproduction of root-knot nematodes (M. arenaria). Eventhough, bacterized and inoculated plants with root-knot nematodes showed better growth than with bacteria untreated plants (induced tolerance). In addition "antibiotica free" culture filtrates (CF) from transitional and stationary bacterial growth phase also promoted reproduction of root-knot nematodes. These CF elicitized tolerance of tomato plants towardsMeloidogyne too. It was proved that B. subtilis could induce a systemic reaction of tomato plants towards root-knot infestation. Besides that test plants treated with B. subtilis (cells) or CF were more attractive to Meloidogyne-Larvae (M.arenaria & M.incognita) than untreated once. CF in 50, 10 and 1% concentrations did not have a nematicidal effect on the root-knot larvae. KNO3 -the carrier of the bacterial preparate (B. subtilis FZB 24®) - had also the same effects on infestation and reproduction of root-knot nematodes. Plant growth was promoted due to application of KNO3. The use of nematode trapping fungus, Arthrobotrys superba gave some range of nematode (root-knot) control (30% gall reduction). Whereas, with the combination ofA. superba and B. subtilis FZB 24® the effect of the fungus was reduced. The application of exogenic phytohormones and precursores showed no effect on plant growth. Reproduction of Meloidogyne was promoted by IAA and combination of IAA and kinetin (not significant). In the tested concentrations of these phytohormones there was no direct mortality effect on root-knot larvae. Content of some enzymes (chitinase, glucanase and peroxidase) from shoot of treated tomato plants was determind. B. subtilis-Isolates (FZB 24® and S18) reduced the population of lesion nematodes,Pratylenchus penetrans in attacked plants (not significant) [9% per root system and 15-20% per g root]. The treatment improved the predisposition of the plants to lesion nematodes (induced resistance). Plant growth was also improved. There was no difference between the bacterial isolates in their effect. KNO3 reduced also nematode population. In general the results would be explained and discussed towards possible mode of actions of rhizobacterium B.subtilis and nematode trapping fungus A. superba.