Thesis (MScAgric)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: Leaf blackening, a postharvest disorder which is characterized by a dark brown to black discoloration, is found in most commercially important Protea cut flower species and cultivars. As this disorder is known to increase with storage time, it is a major concern to the South African industry as the use of sea freight is increasingly preferred due to lower transport costs and a more favourable carbon footprint. The cause of leaf blackening has been strongly linked to a carbohydrate stress exerted by the large inflorescence, thus requiring the utilization of sugar bound polyphenols in the foliage, which when removed, can oxidize enzymatically or non-enzymatically. A study where harvesting was done throughout the season as well as on selected days at 08:00, 12:00, 15:00 and 17:00, concluded that leaf blackening incidences in Protea cv. Sylvia stems varies significantly throughout the season, between years and even with the harvest time of day. Leaf blackening incidences increased from October onwards and remained high until February, before decreasing to acceptably lower levels towards March to May. Carbohydrate- and phenolic content together with water status of leaves at harvest was not able to accurately predict incidence of the associated leaf blackening. However, irrespective of the season of harvesting, leaf blackening was significantly lower when stems were harvested later in the day than compared to stems harvested in the morning. Low sucrose and high water content at these harvest times was positively correlated to high incidences of leaf blackening. In a next study where uptake dynamics of glucose pulsing was investigated, Protea cv. Sylvia was harvested at different times throughout the day, dehydrated to various levels and pulsed with an increasing range of glucose concentrations. Pulsing solution uptake per stem was found to be highly influenced by these factors, as dehydration of stems and a harvest time later during the day both decreased stem water potential, which then increased pulse-solution uptake within a certain time period. The daily harvest time influenced transpiration, whilst pulse-solution uptake decreased with an increase in glucose pulse concentration. When stems were pulsed pre-storage with an increasing range of glucose concentrations, not only did pulses of between 4.7 – 13.7% glucose significantly delayed the incidence of leaf blackening, but it also maintained a positive water balance longer in stems during vase life. Ethanol or acetaldehyde vapour did not provide a viable alternative for reducing leaf blackening incidence in Protea cv. Sylvia, although a synergistic effect was found when ethanol vapour or pulsing was used in combination with glucose. A commercial verification trial disclosed that Protea magnifica and Protea ‘Pink Ice’ reacted more beneficial to ethanol vapour than was observed in ‘Sylvia’. This study confirms that carbohydrate availability within the Protea cut stem remains a key factor in the control of leaf blackening. Factors which assist in maintaining high internal carbohydrate levels, such as enhanced glucose pulse uptake or effective vase solution utilization will contribute to providing an optimum control of leaf blackening during vase life following long-term cold storage. / AFRIKAANSE OPSOMMING: Loofblaarverbruining is ‘n na-oes defek wat gekarakteriseer word deur ‘n donker bruin na swart verkleuring wat voorkom in meeste kommersieël belangrike Protea snyblom spesies en kultivars. Hierdie defek is bekend daarvoor dat dit toeneem met stoortyd, dus is dit ‘n groot kommer vir die Suid-Afrikaanse industrie, met toenemende gebruik van seevrag as vervoer keuse wat laer vervoer kostes en meer gunstige ‘koolstof voetspoor’ bevoordeel. Die oorsaak van loofblaarverbruining word sterk gekoppel aan ‘n koolhidraat stres wat uitgeoefen word deur die groot bloeiwyse op die loofblare, waar suiker-gebonde polifenoliese verbindings ensiematies of nieensiematies geoksideer word met die verwydering van die suiker verbinding. 'n Studie waar geoes was regdeur die seisoen, sowel as op geselekteerde dae om 08:00, 12:00, 15:00 en 17:00, het bevind dat die voorkoms van loofblaarverbruining in stele van Protea kv. Sylvia aansienlik geskil regdeur die seisoen, tussen jare en selfs met die oes tyd gedurende die dag. Die voorkoms van loofblaarverbruining het toegeneem vanaf Oktober en het hoog gebly tot en met Februarie, voordat dit gedaal het tot aanvaarbare laer vlakke teen Maart, tot en met Mei. Koolhidraat-en fenoliese inhoud sowel as die water status van die blare by oes was onsuksesvol om die voorkoms van die gepaardgaande loofblaarverbruining akkuraat te voorspel. Loofblaarverbruining was egter aansienlik laer as stele geoes later in die dag teenoor stele geoes in die oggend, ongeag die seisoen van oes. Lae sukrose en 'n hoë water inhoud geassosieer met hierdie oes-tye was positief gekorreleerd met ‘n hoë voorkoms van loofblaarverbruining. In 'n volgende studie waar die opname dinamika van glukose pulsing ondersoek was, is Protea kv. Sylvia stele geoes op verskillende tye dwarsdeur die dag, gedehidreer tot verskillende vlakke en met 'n toenemende reeks van glukose konsentrasies gepuls. Pulsoplossing opname per steel is sterk beïnvloed deur hierdie faktore, aangesien dehidrasie van die stele asook stele geoes later gedurende die dag die afname van steel waterpotensiaal veroorsaak het, terwyl die puls-oplossing opname versnel het binne ‘n bepaalde tyd. Die tyd van oes beïnvloed ook transpirasie, terwyl vaas oplossing opname afgeneem met 'n toename in glukose puls konsentrasie. Wanneer ‘Sylvia’ stele gepuls was voor stoor met 'n reeks van toenemende glukose konsentrasies, het nie net die puls van tussen 4.7 – 13.7% glukose aansienlik die voorkoms van loofblaarverbruining vertraag nie, maar dit het ook ‘n positiewe water balans langer in stele gedurende die vaas lewe behou. Nie etanol of asetaldehied dampe is bevind as geskikte alternatief vir glukose pulsing om die voorkoms van loofblaarverbruining in Protea kv. Sylvia te verlaag nie, alhoewel ‘n sinergistiese effek waargeneem was wanneer etanol in kombinasie met glukose gebruik was. ‘n Kommersieële bevestigingstoetsing het bevind dat Protea magnifica en ‘Pink Ice’ meer voordeel uit ‘n ethanoldamp behandeling kon trek teenoor ‘Sylvia’. Hierdie studie het bevestig die belangrikheid van koolhidraat beskikbaarheid in die Protea snyblom, vir beheer van loofblaarverbruining. Faktore wat die handhawing van hoë interne koolhidrate vlakke, soos bevorderde glukose puls opname of effektiewe vaas oplossing benutting sal bydra tot ‘n optimal beheer van loofblaarverbruining tydens vaas lewe na langtermyn koueopberging. / National Research Fund (NRF) for their financial support in 2009; Protea Producers of South Africa (PPSA) and Productschap Tuinbouw (PT) as well as the Frank Batchelor Will Trust Grant for the financial support.

Thesis (M. Sc. Agriculture (Plant Protection)) -- University of Limpopo, 2021 / Mimusops zeyheri tree groves made up of seventeen trees collected from communities in Southern Africa were used in this study. The trees had high morphological variations in terms of growth rate, fruit (size and taste) and leaf (shapes and sizes) and their identification was made by communal people from where the trees were collected using their morphological characteristics. Generally, this evergreen tree is drought‒tolerant, salt‒tolerant and highly resistant to root-knot (Meloidogyne spp.) nematodes, along with various other pests. This could probably be attributed to high concentration of latex in aboveground organs. Some typical fungal rust symptoms have been observed believed to be the cause of high leaf abscission in certain groves. Currently, there is no report of leaf rust disease on M. zeyheri plants and the mechanism of resistance to other pests is not documented. The objective of this study was to (i) identify the pathogen associated with M. zeyheri leaf rust symptoms using morphological technique and to (ii) determine levels and types of potential defence chemicals and endophytes in M. zeyheri. Samples of M. zeyheri leaves showing rust like symptoms were collected from University of Limpopo, South Africa (23°53”10’S, 29°44”15’E) during summer in September 2018. Light compound microscope and electron microscope were used in the identification of the leaf rust spores. The species identity of the seventeen M. zeyheri trees that form a grove collection at University of Limpopo was confirmed using internal transcribed spacer (ITS) of ribosomal nuclear DNA. DNA extraction and sequencing was done with the help of Inqaba Biotechnologies. Obtained DNA sequences were aligned using CLUSTALX (2.0), with the phylogenetic tree constructed through the neighbour-joining method (NJM) in MEGA v. 5.1 programme. Evolutionary distances were computed using the Juke– Cantor method. Phytochemicals in leaves were identified and quantified using Liquid chromatography–mass spectrometry (LC–MS) at ARC-VOP. The pustules on the collected leaves contained reddish brown spores. The uredospores were oval and ellipsoidal under a light microscope. The size of spores ranged between 35-37 × 24-26 µm. The cell walls showed bilaminate structures with the outer layer hyaline. The warts were rod shaped with one subequatorial germ pore. The most common identified endophytic fungi observed in all M. zeyheri leaves were Teratosphaeria species, Zeloasperium species, Pezizomycotina. In addition, endophytes such as Cladosporium species, Aspergillus species, Phyllosticta species and Epicoccum species were also identified to be associated with some M. zeyheri trees. There were significant differences on the level of tannins, flavonoids, proteins and phenolics among the M. zeyheri trees. The highest level of tannins was 7.2151 mg/g and the lowest being 2.7232 mg/g. The highest level of flavonoids was 1.1537 mg/g the lowest being 0.0123 mg/g. The level of phenolics among the trees ranged from 2.4749 mg/g to 1.5788 mg/g. Protein content ranged from 5.3100% to 2.7967% among the trees. Very high levels of tannins, flavonoids, phenolics among the trees when compared with others studies indicate the potential role of these metabolites in previously reported resistance of M. zeyheri to a number of pests. The morphological characteristics of the identified leaf rust pathogen causing rust symptoms on M. zeyheri in South Africa is more similar to Maravalia species previously isolated from M. caffra. And as such, this finding is paramount, as it is the first report of association between the pathogen and the plant. Teratosphaeria species has been associated with stem cancer in Eucalyptus trees, its presence in M. zeyheri species in asymptomatic plants is a major find since the tree is common in Limpopo and Mpumalanga, Provinces well known for forestry production making M. zeyheri a potential host for pathogens of commercial forestry production. Other endophytes observed such as Aspergillus species, Phyllosticta species and Cladosporium species have been shown to protect plants against some pests and pathogenic organism. Further studies to determine the direct role of identified phytochemicals and endophytes in the resistance of M. zeyheri to pests needs to be conducted

Thesis (MscAgric (Horticulture))--University of Stellenbosch, 2005. / The occurrence of both pre- and postharvest leaf blackening in certain Protea species and cultivars is a problem that severely limits their marketability, vase life and transport options. This research focuses on : (I) The distribution of carbohydrates in inflorescence bearing stems of certain Protea cultivars from harvest, following pulsing with a 10 g.L-1 glucose solution until four weeks postharvest. Stems were held under a variety of postharvest conditions, and (II) The suppression of Protea postharvest leaf blackening with specific focus on the cultivar ‘Sylvia’ (P. eximia x P. susannae).