Dissertations / Theses on the topic 'Postharvest - Diseases and pests'

Thesis (MEng) -- Stellenbosch University, 2014.
ENGLISH ABSTRACT: As overpopulation threatens the world’s ability to feed itself, food has become an invaluable resource. Unfortunately, almost a third of the food produced for human consumption is lost annually. Pests including insects, phytopathogens and weeds are responsible for more than a third of the annual major crop losses suffered around the world. The majority of current post-harvest control strategies employ synthetic agents. These compounds, however, have been found to be detrimental to the environment as well as human health, which has led researchers to investigate alternative strategies. Biocontrol agents are environmentally compatible, have a lower toxicity and are biodegradable, making them an attractive alternative to the synthetic control agents. The lipopeptides produced by Bacillus spp. in particular, have shown great potential as biocontrol agents against various post-harvest phytopathogens. Most biocontrol strategies apply the biocontrol organism, for example Bacillus, directly, whereas this study focused on the use of the lipopeptide itself as an antifungal agent. This is advantageous as the lipopeptides are less sensitive to their surroundings, such as temperature and pH, compared to living organisms, allowing for the production of a standardized product. This study investigated the production of the Bacillus lipopeptides surfactin, fengycin and iturin under controlled batch conditions. Parameters increasing lipopeptide production were quantified, focussing on antifungal lipopeptides (iturin and fengycin), and lipopeptide production was optimized. Experiments were performed in a fully instrumented 1.3 L bench-top bioreactor and lipopeptide analyses were performed via high pressure liquid chromatography (HPLC) and liquid chromatography-mass spectroscopy (LC-MS). After screening four Bacillus spp., Bacillus amyloliquefaciens DSM 23117 was found to be the best antifungal candidate. This was based on it outperforming other candidates in terms of maximum antifungals produced, Yp/x,antifungals (yield per cells), and antifungal productivity. Nitrate, in the form of NH4NO3, was critical for lipopeptide production and an optimum concentration was observed above which the CDW (cell dry weight) no longer increased significantly and both μmax (maximum specific growth rate, h-1) and lipopeptide production decreased. For μmax, the optimum NH3NO4 concentration was 10 g/L and for lipopeptides it was 8 g/L. At these respective NH4NO3 concentrations μmax = 0.58 (h-1), the maximum antifungals (fengycin and iturin) were 285.7 mAU*min and the maximum surfactin concentration was 302 mg/L. The lipopeptides produced by B. amyloliquefaciens, the antifungals (fengycin and iturin) and surfactin, are secondary metabolites, regardless of the optimization treatment, i.e. increased NH4NO3 concentrations. Using 30% enriched air extended the nitrate utilization period, suggesting that when increasing supply concentration, more oxygen was available to act as electron acceptors, allowing nitrate to be used for lipopeptide production. The number of iturin and fengycin homologues generally increased with an increase in nitrate concentration. This suggested that process conditions, such as nitrate concentration, can be used to manipulate homologue ratios, allowing for the possibility to tailor-make biocontrol-agent upstream, during the production process, and possibly increase the efficacy of the biocontrol strategy. The lipopeptides produced by B. amyloliquefaciens showed complete inhibition against Botryotinia fuckeliana and diminished the growth capabilities of Botrytis cinerea. No inhibition was observed against Penicillium digitatum. These results indicate potential of the biocontrol strategy, although scale-up and fed-batch studies are recommended, especially when considering commercial implementation. Studies regarding the lipopeptide application method, i.e. a single application or multiple applications, should also be investigated as this will influence the efficacy of the lipopeptides against the target organisms.
AFRIKKANSE OPSOMMING: Met oorbevolking wat die wêreld se vermoë om die groeiende bevolking te onderhou belemmer, het dit noodsaaklik geword om huidige voedselbronne te beskerm. Daar word beraam dat een derde van die voedsel wat wêreldwyd geproduseer word vir menslike verbruik verlore gaan elke jaar. Verder is insekte, plantpatogene en onkruide verantwoordelik vir meer as ‘n derde van die verliese rakend jaarlikse oeste. Meeste bestaande na-oes beheermetodes maak gebruik van sintetiese stowwe. Ongelukkig kan hierdie verbindings nadelig wees vir die omgewing sowel as menlike gesondheid. Navorsers het hulsef dus toespits daarop om alternatiewe beheermetodes te ondersoek. Bio-beheermetodes is omgewingsvriendelik sowel as bio-afbreekbaar, wat hulle ideale alternatiewe maak vir die sintetiese stowwe. Bacillus spp. lipopeptiede het veral hoë potensiaal getoon as bio-beheermiddels teen verskeie na-oes plantsiektes. Meeste bio-beheermetodes wend die biobeheer organisme, soos Bacillus, direk aan, waar hierdie studie op die gebruik van lipopeptiede as ‘n beheermiddel gefokus het. Die voordeel is dat lipopeptidiede minder sensitief is vir hul omgewings, soos temperatuur en pH, i.v.m. organismes en die moontlikheid bied van ‘n gestandardiseerde produk. Hierdie studie het die produksie van spesifieke Bacillus lipopeptide, naamlik surfactin, fengycin en iturin, onder beheerde lottoestande ondersoek. Parameters wat lipopeptied produksie verhoog is gekwantifiseer, spesifiek antifungiese middels (iturin en fengycin) en lipopeptied produksie is geoptimeer. Eksperimente is uitgevoer in ‘n 1.3 L bioreaktor en lipopeptiedanaliese is met behulp van hoë druk vloeistof chromatografie en vloeistofchromatografie-massa spektroskopie uitgevoer. Van die vier moontlike Bacillus spp., was Bacillus amyloliquefaciens DSM 23117 die mees belowende antifungus-produserende kandidaat. Dit het beter resultate gelewer in terme van maksimale antifungiese produksie, Yp/x,antifungies (opbrengs per sel) asook antifungiese produktiwiteit. Nitraat, in hierdie geval NH4NO3, was noodsaaklik vir lipopeptied produksie en ‘n optimale konsentrasie is waargeneem waarbo die seldigtheid nie meer beduidend toegeneem het nie en beide die μmax (maksimale spesifieke groei tempo, h-1) en lipopeptied produksie afgeneem het. Die optimale NH4NO3 konsentrasie vir μmax was 10 g/L en vir lipopeptiedproduksie was 8 g/L. By 10 g/L NH4NO3 was μmax = 0.58 (h-1) en by 8 g/L was die maksimale antifungiese produksie (fengycin en iturin) 285.7 mAU*min en die maksimale surfactin produksie 302 mg/L onderskeidelik. Die lipopeptide, die antifungiese middels (fengycin en iturin) en surfactin, geproduseer deur B. amyloliquefaciens is sekondêre metaboliete, ongeag van die optimerings-behandelinge wat toegepas word, soos ‘n verhoging in NH4NO3 konsentrasie. Die gebruik van 30% verrykte suurstof het die nitraat verbruikingsperiode verleng, wat voorgestel het dat met die verryking, meer suurstof beskikbaar was om te dien as finale elektron ontvanger en sodoende die nitraat beskikbaar te stel vir lipopeptied produksie. Iturin en fengycin homoloë, oor die algemeen, het toegeneem soos wat die nitraat konsentrasie verhoog is. Hierdie resultate dui daarop dat prosestoestande, soos nitraat konsentrasie, gebruik kan word om die verhouding waarin lipopeptied homoloë geproduseer word te manipuleer. Hierdie resultate dui op die potensiaal vir die stroomop produksie van ‘n unieke bio-beheermiddel, wat die effektiwiteit van die bio-beheermetode moontlik sal verhoog. Die geproduseerde lipopeptiede het totale inhibisie getoon teen Botryotinia fuckeliana en ook fungiese aktiwiteit belemmer met Botrytis cinerea. Geen inhibisie is getoon teen Penicillium digitatum nie. Hierdie resultate toon die potensiaal van die bio-beheermetode, maar ‘n opskalerings-studie asook ‘n voerlot studie word aanbeveel, veral met die oog op moontlike kommersiële implementering van die strategie. Verdere studies met betrekking tot die aanwendingsmetode van die lipopeptiede moet ook verder ondersoek word, m.a.w. enkel teenoor menigte aanwendigs, aangesien dit die effektiwiteit van die lipopeptiede teen die teikenorganismes sal beïnvloed.

Orientador: Benedito Carlos Benedetti
Texto em português e Inglês
Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Agrícola
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Resumo: Botrytis cinerea Pers, causador da doença conhecida como mofo cinzento, é o principal problema para a conservação pós-colheita de uvas de mesa. A utilização do dióxido de enxofre (SO2) é a prática pós-colheita mais comum para o controle desta doença. Pesquisas buscam alternativas a este produto devido às reações que causa em pessoas alérgicas, danos que pode causar nos frutos e às restrições ao seu uso em sistemas de produção orgânico. Foram avaliados os efeitos da aplicação de uma atmosfera de 40% de CO2 por 24 ou 48 horas (pré-armazenagem) combinado com armazenagem em atmosfera controlada (AC) (12% O2 + 12% CO2) no controle de B. cinerea, e nos atributos de qualidade de uvas 'Flame Seedless' e 'Crimson Seedless'. Também foram avaliados, em uvas 'Crimson Seedless', e os efeitos da associação deste tratamentos com aplicações pré-colheita de cloreto de cálcio (CaCl2) ou dióxido de cloro (ClO2). Os tratamentos foram aplicados em uvas orgânicas infectadas de três formas: infectadas naturalmente, superficialmente inoculadas com conídios e inoculadas com uma baga coberta de micélio. Uvas 'Flame Seedless', naturalmente infectadas, tratadas com 40% de CO2 por 48 horas + AC apresentaram redução da podridão pós-colheita, de 22% para 0,6%, após 4 semanas, e de 100% para 7,4%, após 7 semanas. O pré-armazenamento em 40% de CO2 sozinho também limitou a incidência de mofo cinzento em frutos infectados naturalmente e em uvas inoculadas artificialmente, porém foi menos eficaz do que quando seguido pelo armazenamento em AC. A aplicação de CaCl2 ou ClO2 em pré-colheita reduziu a incidência do mofo cinzento em uvas 'Crimson Seedless' inoculadas com uma solução de conídios, porém não houve controle quando os cachos de uva foram inoculados com micélio. A aplicação de CaCl2 e ClO2 reduziram o mofo cinzento de 45% para 23,2% e 15.6%, respectivamente, em cachos inoculados com conídios e avaliados após 6 semanas armazenadas 0oC. O pré-tratamento com 40% CO2 + CA não afetou as características físico-químicas e sensoriais de uvas 'Crimson Seedless' ou 'Flame Seedless'. Em experimentos in vitro os tratamentos com 40% CO2 por 24 ou 48 h limitaram o crescimento micelial até 72 horas após o tratamento. A germinação dos conídios de B. cinerea foi retardada por 12h. Os resultados mostram que o pré-tratamento com 40% CO2 + CA possui grande potencial para ser adotado como prática comercial para conservação de uvas orgânicas
Abstract: Gray mold, caused by Botrytis cinerea Pers, is the main postharvest decay of table grapes. The use of sulfur dioxide (SO2) is the common post-harvest practice for its control. Several researchers are looking for alternative methods of control, because SO2 can cause allergic reactions, damage fruits and also it cannot be applied in organic production system. In this thesis, it was evaluated the effects of applying an atmosphere of 40% CO2 for 24 or 48 hours (pre-storage) combined with controlled atmosphere storage (CA = 12% O2 + 12% CO2) in the control of B. cinerea, and the effects in the quality and sensory attributes of 'Flame Seedless' and 'Crimson Seedless' table grapes. In addition, it was evaluated the efficacy of CaCl2 or ClO2 application in pre-harvest alone or in combination with pre-storage of 40% CO2 for 24 h + CA, to control gray mold on 'Crimson Seedless' table grapes, and the determination of the impact of these treatments on fruit quality. The treatments were applied in certified organic table grapes naturally infected, surface inoculated and nesting inoculated (inoculated with an infected berry). After 4 weeks of storage, the pre-storage in 40% CO2 for 48 hours + CA reduced postharvest rot from 22% to 0.6%, and after 7 weeks, the decay was reduced from 100% to 7.4% in 'Flame Seedless' naturally infected. The pre-storage in 40% CO2 alone also reduced the incidence of gray mold in fruits naturally infected and in artificially inoculated, but it was less effective than combined treatment. The application of CaCl2 or ClO2 pre-harvest reduced the incidence of gray mold on grapes 'Crimson Seedless' inoculated with a spore solution, but there was no control when fruits were nesting inoculated. After 6 weeks at 0oC, the application of CaCl2, and the ClO2 in fruits surface inoculated, reduced the gray mold from 45% to 23.2% and 15.6%, respectively. The pretreatment with 40% CO2 + CA did not affect quality and nor sensory attributes for both varieties tested. In vitro experiments, the treatment with 40% CO2 for 24 or 48 h limited mycelial growth for at least 72 hours after treatment. Conidial germination of B. cinerea was delayed for 12 hours. Our results showed the potential that pre-treatment with 40% CO2 associated with CA has to be adopted in commercial practice for preservation of organic grapes
Doutorado
Tecnologia Pós-Colheita
Doutor em Engenharia Agrícola

Thesis (MScAgric)--Stellenbosch University, 2013.
ENGLISH ABSTRACT: The export of South African fruit to some of its biggest international markets may be rejected if the phytosanitary pest, Phlyctinus callosus (Coleoptera: Curculionidae) is found in fruit consignments. An alternative to methyl bromide fumigation is needed and one of the most promising of the alternative treatments is phytosanitary irradiation as it is environmentally friendly, does not leave residues on food or in the environment and it is effective against a wide variety of insects. Field-collected weevils were treated with five doses of gamma irradiation (5, 10, 20, 40 and 80 Gy) and the fecundity and fertility of mating crosses of treated males and females with treated and untreated individuals of the opposite sex, were determined to evaluate the effect on P. callosus reproductive ability post-treatment. Results indicated that irradiation treatment did not affect fecundity, but fertility was significantly affected, decreasing as the irradiation dose increased. Females were more susceptible to the irradiation treatment than males, and after treatment with 80 Gy, eggs laid by females and mated with either treated or untreated males, did not hatch. A generic dose of 400 Gy for all insect pests except tephritid fruit flies and pupae and adult Lepidoptera is currently approved by USDA-APHIS (United States Department of Agriculture – Animal and Plant Health Inspection Services) for use on certain commodities. Results from the present study support the development of a species-specific dose for P. callosus, as well as the development of a group generic dose for the Curculionidae that is lower than 400 Gy. Effective phytosanitary irradiation treatments are only feasible if the treatment does not adversely affect fruit quality and the marketability of export fruit. Therefore, an investigation of the effects of irradiation disinfestations treatments on the quality of the new pluot cultivar, ‘Flavor Fall’ was made. Packed cartons were treated with three doses of gamma irradiation: 400 Gy, 900 Gy and 1400 Gy. After treatment fruit underwent a PD 7 dual temperature cold storage regime for 42 days and a shelf-life simulation for 7 days. The impact of insect-proof bags, sometimes required by importing countries to keep insects off packaged fruit, was also investigated. Respiration rate of the fruit was measured throughout and fruit quality evaluations were done after cold storage and after shelf-life. The results indicated that quality parameters measured at the end of cold storage, which would be after the fruit arrives at the export markets, were above the minimum standards for overseas markets. Gel breakdown was unacceptably high after the higher temperature exposure of shelf-life for fruit treated with the 900 and 1400 Gy doses. The insect-proof bags reduced shrivel, but resulted in higher incidence of gel breakdown. The use of irradiation, together with the use of the insect-proof bag, has potential as an alternative postharvest mitigation treatment for plums. Lastly, an investigation into potential rearing methods for P. callosus, including recommendation for the future, was made as the availability of a sustainable rearing method that ensures a consistent supply of high quality P. callosus adults would enable continuous research with greater numbers of this pest. The information generated in this study provides a greater understanding of the radiation biology of, not only this curculionid species, but the Curculionidae as a group, and is valuable in advancing the development of alternative postharvest control measures against this phytosanitary pest.
AFRIKAANSE OPSOMMING: Suid Afrikaanse vrugte uitvoere na van die grootste internasionale markte mag weg gewys word as die fitosanitêre pes, Phlyctinus callosus (Coleoptera: Curculionidae) in die versending gevind word. ‘n Alternatief vir metiel bromied beroking word benodig en een van die mees belowende alternatiewe behandelings is fitosanitêre bestraling aangesien dit omgewings vriendelik is, nie residue op kos of in die omgewing los nie, en effektief is teen ‘n wye verskeidenheid van insekte. Veldversamelde kalanders is behandel met vyf dosisse gamma bestraling (5, 10, 20, 40 en 80 Gy) waarna die vrugbaarheid van paringskruisings bepaal is deur kruisings tussen behandelde manlike en vroulike kalanders met behandelde en nie-behandelde individue van die teenoorgestelde geslag te maak, en so die na-behandelings effek op die voortplantings vermoeë van P. callosus te evalueer. Die resultate het getoon dat die bestralings behandeling geen invloed gehad het op die hoeveelheid eiers wat gelê is nie, maar dat die uitbroei van eiers aanduidend geaffekteer is deur die behandeling. Die hoeveelheid eiers wat uitgebroei het, het minder geraak soos die bestralings behandeling toegeneem het. Vroulike kalanders was meer sensitief vir die behandeling en na 80 Gy, of hul gekruis is met behandelde of niebehandelde mannetjies, het geen eiers uitgebroei nie. ‘n Generiese dosis van 400 Gy vir alle insekte, uitsluitend tephritiese vrugte vlieë en papies en volwasse Lepidoptera is huidiglik goedgekeur deur die USDA-APHIS (United States Department of Agriculture – Animal and Plant Health Inspection Services) vir sekere kommoditeite. Die resultate van die huidige studie ondersteun die ontwikkeling van ‘n spesie-spesifieke dosis vir P. callosus, so ook die ontwikkeling van ‘n generiese groep dosis vir Curculionidae wat laer as 400 Gy is. Effektiewe fitosanitêre bestralings behandeling is slegs moontlik indien die behandelings dosis nie nadelig vir vrugkwaliteit en die bemarking van uitvoer vrugte is nie. Dus is die effek wat bestralings bestryding behandeling op die kwaliteit van ‘n nuwe pluot kultivar, ‘Flavor Fall’ ondersoek. Vrugte verpak in kartonne is met drie dosisse gamma bestraling behandel: 400 Gy, 900 Gy en 1400 Gy. Na behandeling is die vrugte deur ‘n PD 7 dubbel temperatuur koelopbergings regime van 42 dae en rak-lewe simulasie vir 7 dae gesit. Die impak van insek-bestande sakke wat insekte van die verpakte vrugte weg hou en soms deur invoerende lande ‘n vereiste is, is ook ondersoek. Respirasie tempo van die vrugte is getoets en vrugkwaliteit evaluasies is gedoen na koelopberging en rak-lewe. Die resultate het getoon dat die kwaliteits maatstawwe wat getoets is na koelopberging (wat tipies is wanneer die vrugte by die uitvoer mark arriveer), almal bo die minimum standaarde van die uitvoer markte was. Gel-afbraak was onaanvaarbaar hoog na blootstelling aan die hoër temperature tydens rak-lewe vir vrugte wat behandel is met 900 en 1400 Gy. Die insek-bestande sakke het verrimpeling verminder, maar die voorkoms van gel-afbraak vermeerder. Die gebruik van bestraling, tesame met die insek-bestande sakke, het potensiaal as alternatiewe na-oes behandeling vir pruime. Laastens is ‘n ondersoek ingestel vir moontlike teeltegnieke vir P. callosus en aanbevelings gemaak vir toekomstike studies. Die beskikbaarheid van ‘n volhoubare teeltegniek wat konstante, hoë kwaliteit P. colossus individue verskaf sal aaneenlopende navorsing met groter getalle van die pes moontlik maak. Die inligting wat deur hierdie studie gegenereer is help om die bestralings biologie, nie net van hierdie curculionid spesie nie, maar die Curculionidae as ‘n groep te verstaan, en is kosbaar in die bevordering van ontwikkeling van alternatiewe na-oes beheer meganismes teen hierdie fitosanitêre pes.

Stony corals (Order Scleractinia) are susceptible to a variety of diseases, which can occur from abiotic or biotic factors, or a combination of both. Public aquaria provide opportunities to study coral disease. Because coral mucus is the first line of defense against disease, the Biolog EcoplateTM is a useful tool to detect differences in microbial assemblages in the surface mucopolysaccharide (mucus) layer when comparing healthy and diseased corals. Histological examination is essential to document structural changes in coral tissue in response to diseases. This study identifies and characterizes diseases in captive corals through visual recognition, characterization of carbon utilization by microbial assemblages in coral mucus samples, and histological examination. In March 2010, surveys were sent to public aquaria throughout the United States that house corals. If the survey was returned indicating that the aquarium had diseased coral specimens present, sample kits were sent to the aquarium to acquire photographs, mucus samples for microbial carbon utilization analysis, and tissue samples for histological examination. Eighteen aquaria participated in the survey and 25 sets of samples were provided. The gross lesions from diseased samples fit into six categories: discoloration associated with darkening of the tissue or with color loss (bleaching), growth anomalies, and tissue loss associated with pests, with brown jelly, or with no obvious cause. Seven categories of possible contributing factors were reported: addition of inadequately quarantined corals to the tank, damage during transport, change of location, manual stress, and variations in light, salinity, or temperature. Introduction of inadequately quarantined specimens was the most common possible contributing factor to pest introduction. Significant differences in carbon source utilization were found between tank-water samples and mucus from both healthy and diseased areas of sampled corals. Although mucus samples from healthy and diseased samples did not differ in carbon source utilization overall, D-mannitol was used by 52% of microbial assemblages from mucus from diseased areas compared with only 17% of microbial assemblages from healthy mucus samples. Histologically, the most commonly observed features across all samples were healthy zooxanthellae, endolithic organisms, and nematocysts, all of which are normal features that can be influenced by stress factors. Brown granular material and ciliates were found associated with some anomalies, primarily the three categories of tissue loss. The presence of dense aggregates of zooxanthellate-engorged ciliates in corals afflicted with brown jelly was highly similar to histological observations of brown band syndrome, previously described from natural coral reefs.

[EN] Citrus spp. (Rutaceae) are the most widely produced fruits and they are grown in over a hundred countries. During postharvest, major losses are caused by fungal diseases, that for decades have been controlled by treatments with synthetic fungicides. However, their use is continuously more restricted by distributors and supermarket chains, which lower the Maximum Residue Level (MRL) to one third of the authorized or even ban the use of certain fungicides. This has loss of efficacy of treatments and development of resistant strains of pathogens. Alternatives are necessary to substitute or combine the use of synthetic fungicides. The objective of the present work was to evaluate alternative treatments against green and blue molds, caused by Penicillium digitatum (Pers.: Fr.) Sacc. and Penicillium italicum Wehmer, respectively. Food additives and GRAS substances were tested in in vivo preliminary tests against green and blue molds. Potassium sorbate (PS) (Chapter 2) and sodium benzoate (SB) (Chapter 1) were the most effective. 60-s dips in 3% (w/v) SB above 50°C reduced by 90% green and blue mold incidence on 'Valencia' oranges after 7 days at 20°C. This treatment was also effective on 'Lanelate' oranges, 'Fino' lemons and 'Ortanique' mandarins, but not on 'Clemenules' mandarins. Heated solutions combining SB with low doses of imazalil (IMZ) were synergistic. In semi-commercial trials with 'Marisol' Clementine mandarins, the combination PS+IMZ at 20°C allowed a significant reduction of the IMZ doses still controlling green mold. A new colorimetric method that employed extraction of PS from macerated fruit, followed by reaction with 2-thiobarbituric acid (Chapter 3) showed that in oranges treated with PS and stored at 15°C, residues declined initially rapidly and later more slowly, until residues stopped declining after 6 d. A brief double-dip rinse in tap water, immediately after immersion of lemons in a 2% (w/v) PS, removed more than 90% of PS residue. Exposure at 33ºC with 15 kPa CO2 for 24 h or 30 kPa O2 for 48 h effectively controlled both green and blue molds after 7 days of incubation at 20ºC, but control of both diseases lowered after 15 days (Chapter 4). Combining PS dip treatments with gas exposure showed a synergistic effect, that maintained the efficacy of combined treatments during 15 days. In cold storage, after 14, 28, and 42 days, green mold incidence on 'Clemenules' and 'Ortanique' mandarins treated with PS dips and 30kPa O2 for 48h was reduced by 100, 96 and 68%, and 100, 97 and 79%, respectively. Again, a synergistic effect between treatments was observed (Chapter 5). Postharvest green mold and blue molds were effectively controlled by fumigation of lemons and oranges for 6 h at 22ºC with two applied dosages of 3000 ¿L L-1 of ammonia that was injected initially and again 2 h later (Chapter 6). Ammonia fumigation controlled an IMZ-resistant isolate of P. digitatum. When fruit were first immersed in 10 or 30 mg L-1 IMZ (about 10% of typical commercial rates) before ammonia fumigation, a single fumigation with 1500 ¿L L-1 of ammonia was adequate to control both diseases and the increase in effectiveness was additive or synergistic. All these treatments, at different degrees, could replace synthetic fungicides or augment IMZ performance in citrus postharvest decay management. The development of green and blue molds on the most commercially important citrus species and cultivars was assessed for four consecutive harvest seasons and associated with fruit quality parameters (Chapter 7). Fruit susceptibility of the cultivars to green mold in increasing order was Nova, Sanguinelli, Ortanique, Lanelate, Navelate, Fortune, Clemenules, Valencia, W. Navel, Oronules, Clemenpons, Marisol, Salustiana, Hernandina, and Nadorcott. Peel characteristics were the most influencing quality parameters on the susceptibility of citrus fruit to green and blue molds.
[ES] Los cítricos (Citrus spp. fam. Rutaceae) son la fruta más consumida a nivel mundial y se cultivan en más de un centenar de países. Durante el manejo en poscosecha, la mayor parte de las pérdidas se deben a enfermedades fúngicas, que durante décadas se han controlado mediante tratamientos con fungicidas de síntesis. Sin embargo, su uso está continuamente más restringido por parte de los distribuidores y cadenas de supermercados, que reducen el Límite Máximo de Residuos (LMR) a un tercio del establecido o incluso prohíben el uso de ciertos fungicidas. Esto ha provocado la pérdida de eficacia de los tratamientos y el desarrollo de cepas de patógenos resistentes a los fungicidas. Se necesitan pues alternativas para sustituir o combinar con el uso de fungicidas de síntesis. El objetivo de este trabajo fue evaluar tratamientos alternativos contra las podredumbres verde y azul, causadas por Penicillium digitatum (Pers.: Fr.) Sacc. y Penicillium italicum Wehmer, respectivamente. Aditivos alimentarios y sustancias GRAS se probaron en test in vivo preliminaries contra los mohos verde y azul. El benzoato sódico (BS; Capítulo 1) y el sorbato potásico (SP; Capítulo 2) fueron los más eficaces. Baños de 60 s en 3% (p/v) BS sobre 50°C redujeron en un 90% la incidencia de las podredumbres verde y azul en naranjas 'Valencia Late' tras 7 días a 20°C. Estos tratamientos fueron también efectivos en naranjas 'Lanelate', limones 'Fino' y mandarinas híbridas 'Ortanique', pero no en mandarinas 'Clemenules'. La combinación de baños calientes de BS con dosis bajas de imazalil (IMZ) fueron sinérgicas. Un nuevo método colorimétrico que empleó la extracción del SP de la fruta macerada, seguido de una reacción con ácido 2-tiobabitúrico (Capítulo 3) mostró que en naranjas tratadas con SP almacenadas a 15°C, los residuos descendieron rápidamente al inicio y lentamente más tarde, hasta que dejaron de descender tras 6 días. Un doble aclarado rápido con agua corriente inmediatamente después de la inmersión de limones en SP al 2% (p/v), eliminó más del 90% del residuo de SP. Los choques gaseosos a 33ºC y 15kPa CO2 durante 24 h o 30 kPa O2 durante 48 h controlaron de manera eficaz las podredumbres verde y azul tras 7 días de incubación a 20ºC, pero el control de las dos podredumbres descendió tras 15 días (Capítulo 4). En almacenamiento en frío, tras 14, 28 y 42 días, la incidencia de moho verde en mandarinas 'Clemenules' y 'Ortanique' tratadas con baños con SP y 30kPa O2 durante 48h se redujo en un 100, 96 y 68%, y 100, 97 y 79%, respectivamente. De nuevo, una fuerte sinergia se observó entre los tratamientos (Capítulo 5). Las podredumbres verde y azul se controlaron eficazmente mediante la fumigación de limones y naranjas durante 6 h a 22ºC con dos dosis de 3000 ¿L L-1 amoniaco, inyectadas al inicio del tratamiento y 2 h después (Capítulo 6). El tratamiento con amoniaco controló una cepa de P. digitatum resistente a IMZ. Cuando la fruta se bañó en 10 o 30 mg L-1 IMZ (un 10% de la dosis comercial) antes de la fumigación con amoniaco, una sola aplicación con 1500 ¿L L-1 amoniaco fue suficiente para controlar las dos podredumbres, y el incremento en la eficacia fue aditivo o sinérgico. Todos estos tratamientos, a diferentes niveles, podrían sustituir a los fungicidas de síntesis o aumentar la eficacia de los tratamientos con IMZ en el manejo poscosecha de los cítricos. El desarrollo de las podredumbres verde y azul en las especies y variedades de cítricos más importantes comercialmente se evaluó durante cuatro campañas consecutivas y se relacionó con los parámetros de calidad de la fruta (Capítulo 7). La susceptibilidad de los cultivares en orden creciente fue Nova, Sanguinelli, Ortanique, Lanelate, Navelate, Fortune, Clemenules, Valencia, W. Navel, Oronules, Clemenpons, Marisol, Salustiana, Hernandina, y Nadorcott. Las características de la corteza fueron los parámetros de calidad
[CAT] Els cítrics (Citrus spp. fam. Rutaceae) son la fruita més consumida a nivell mundial i es cultiven en un centenar de països. Durant la poscollita, la major part de les pèrdues es deuen a malalties fúngiques, que durant dècades s'han controlat mitjançant tractaments amb fungicides de síntesi. No obstant, l'ús d'aquestes pràctiques està contínuament més restringit per part dels distribuïdors y cadenes de supermercats, que redueixen el Límit Màxim de Residus (LMR) a un terç del permès per la legislació o inclús prohibeixen l'ús d'alguns fungicides. Açò ha provocat la minora de l'eficàcia dels tractaments poscollita i el desenvolupament de soques dels patògens resistents als fungicides. Es necessari per lo tant trobar i implementar alternatives per a substituir o combinar l'ús de fungicides de síntesi. L'objectiu d'aquest treball ha sigut avaluar tractaments alternatius contra les podridures verda i blava, causades per Penicillium digitatum (Pers.: Fr.) Sacc. i Penicillium italicum Wehmer, respectivament. Additius alimentaris i substàncies GRAS es provaren en test in vivo preliminars contra les podridures verda i blava. El benzoat sòdic (BS; Capítol 1) i el sorbat potàsic (SP; Capítol 2) foren els més eficaços. Solucions calentes de BS combinades amb baixes dosis d'imazalil (IMZ) foren sinèrgiques. Esta combinació va reduir l'incidència de les podridures verda i blava en quasi el 100% en taronges 'Valencia Late' després de 8 setmanes a 5°C i 7 dies a 20°C. En assajos semicomercials mandarines 'Marisol', la combinació SP+IMZ a 20°C va permetre una baixada significativa de la dosis de IMZ sense perdre eficàcia contra la podridura verda. Un nou mètode colorimètric que inclou l'extracció del SP de la fruita macerada, seguit d'una reacció amb àcid 2-tiobabitúric (Capítol 3) mostrà que en taronges tractades amb SP conservades a 15°C, els residus descendiren ràpidament al inici i lentament mes tard, fins que deixaren de descendir després de 6 dies. Un doble rentat ràpid amb aigua corrent immediatament després de d'immersió de llimones en SP al 2% (p/v), eliminà més del 90% del residu de SP. Els xocs gasosos a 33ºC i 15kPa CO2 durant 24 h o 30 kPa O2 durant 48 h controlaren de manera eficaç les podridures verda i blava després de 7 dies d'incubació a 20ºC, però el control de les dos podridures descendí després de 15 dies (Capítol 4). La combinació de banys amb SP i xocs gasosos mostrà un efecte sinèrgic, que mantingué l'eficàcia dels tractaments combinats durant 15 dies. En conservacions en fred, després de 14, 28 i 42 dies, d'incidència de podridura verda en mandarines 'Clemenules' i 'Ortanique' tractades en banys amb SP i 30kPa O2 durant 48h es reduí en un 100, 96 i 68%, i 100, 97 i 79%, respectivament. De nou, una forta sinergia s'observà entre els tractaments (Capítol 5). Les podridures verda i blava es controlaren eficaçment mitjançant la fumigació de llimones i taronges durant 6 h a 22ºC amb dos dosis de 3000 ¿L L-1 amoníac, injectades al inici del tractament i 2 h després (Capítol 6). El tractament amb amoníac controlà una soca de P. digitatum amb un alt nivell de resistència a IMZ. Tots estos tractaments, a diferents nivells, podrien substituir als fungicides de síntesis o augmentar l'eficàcia dels tractaments amb IMZ en el maneig poscollita dels cítrics. El desenvolupament de les podridures verda i blava en les especies y varietats de cítrics més importants comercialment s'avaluà durant quatre campanyes consecutives i es relacionà amb els paràmetres de qualitat de la fruita (Capítol 7). La susceptibilitat dels distints cultivars en ordre creixent fon Nova, Sanguinelli, Ortanique, Lanelate, Navelate, Fortune, Clemenules, Valencia, W. Navel, Oronules, Clemenpons, Marisol, Salustiana, Hernandina, i Nadorcott. Les característiques de la pell foren els paràmetres de qualitat més influents en la susceptibilitat dels cítrics a les pod
Montesinos Herrero, CI. (2016). Evaluation of alternatives to synthetic fungicides to control citrus postharvest diseases [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/63662
TESIS

Doctor of Philosophy (PhD)
The goal of this research was to test commercially viable alternative methods to fungicides for controlling storage diseases of melons that are safe for human health and the environment. Initially, experiments were conducted on melons to develop a protocol for optimum conditions of disease development during storage for different pathogens and for different stages of fruit maturity. For all pathogens tested, the study found that humidity greater than 90% and temperatures above 20° C support infection and rapid growth of disease. Differences in the rate of infection and extent of disease development after the inoculation of different storage pathogens was observed between rockmelon and honeydew melons, indicating differences in host pathogen interactions. Among the tested pathogens, Alternaria spp. was the least aggressive in infection and disease severity, growing considerably slower than Fusarium acuminatum which was moderately aggressive and Rhizopus spp. which was very aggressive, in comparison. Green half-slip melons showed greater resistance to pathogen attack than green full-slip fruit, while yellow full-slip melons were highly susceptible to pathogen attack. Therefore, the laboratory experiments for postharvest treatments of rockmelons were performed using green full-slip fruit challenged with F. acuminatum. Evaluation of physical and safe chemical methods of postharvest treatment to control postharvest diseases of melons showed that none of the treatments alone was as effective as the commercially available fungicide. Hot water solutions of safe compounds considerably increase their efficacy against postharvest rots, however, symptoms of phytotoxicity on the rind after dipping made their use unacceptable. Iodine was the only safe chemical tested which did not cause any phytotoxicity on melons. When combined with hot water, iodine showed the best control of storage rots and was as good as the fungicides carbendazim or guazatine. Hot water iodine dipping of fruit also delayed ripening and fruit were firmer during storage for a longer period of time. Systemic acquired resistance (SAR) was evaluated as a method of controlling powdery mildew in glasshouse grown rockmelon seedlings by treating with the activators 2,6-dichloroisonicotinic acid (INA) or benzothiadiazole (BTH) or water. Increased resistance due to application of INA or BTH, was observed by the reduction of powdery mildew on pre-inoculated detached leaves and also on intact leaves from natural infections. Heightened resistance due to spraying with elicitors of SAR, was further evident by the increased activities of the pathogenesis related proteins (PR proteins), peroxidase and accumulation of phenolics or antifungal compounds during and after challenge inoculation. Field grown rockmelons were treated with INA or BTH or BABA (β-aminobutyric acid) or water at various stages of plant growth and evaluated for increased resistance against pre and postharvest diseases. Both powdery mildew and downy mildew were significantly less on the SAR elicitor treated plants. Preharvest treatment with SAR elicitors also reduced storage diseases of the harvested rockmelon fruit. The reduction in postharvest disease was similar whether plants were treated once, three weeks before harvest, or given four sprays during the growing season beginning at anthesis. A further postharvest dip with 500 ppm of guazatine gave substantial reduction of storage rots of melons. Enhanced activities of chitinase and peroxidase, two major PR-proteins, compared to the control, indicated induction of defence had occurred in the foliage and fruit as a result of SAR. Over the course of four field and one glasshouse experiments slight phytotoxicity was observed in plants frequently sprayed with INA or BTH, but no phytotoxicity was seen after a single spray during the late stages of fruit development. The combination of SAR elicitor treatment and use of a safe postharvest dip provided substantial control of storage rots of rockmelons. The best treatment for control of storage rots involved application of BTH (50 ppm) two weeks before harvest, combined with a hot iodine dip (55° C) of fruit, achieving equivalent or better disease control than use of guazatine fungicide dip.

Dissertation (PhD (Agric))--University of Stellenbosch, 2002.
ENGLISH ABSTRACT: Fungi belonging to the genus Botryosphaeria are heterotrophic micromycetes that can be pathogens on woody plants. They cause serious, and in some cases devastating losses to crops through leaf necrosis, stem cankers and plant death. The Proteaceae cut-flower industry in South Africa accounts for 70% of the national cut-flower enterprise. Botryosphaeria diseases are a major impediment to production and trade of Proteaceae and there is an urgent need to investigate the etiology, epidemiology and control of these diseases. Losses of one of the most important proteas, P. magnifica, amount to 50% or more, locally. The main aims of this study were therefore to establish the etiology and aspects of epidemiology of Botryosphaeria stem cankers on P. magnifica and other Proteaceae, and to investigate methods of disease control. Although there is a vast body of information pertaining to this fungus, which was reviewed in Chapter 1, there is relatively little information available on Botryosphaeria on Proteaceae. The taxonomy of Botryosphaeria requires thorough review, and molecular techniques need to be employed to resolve species identities. In Chapter 2, it was found that Phyllachora proteae, a leaf pathogen of proteas, produced a Fusicoccum anamorph, which is described as F. proteae. A sphaeropsis-like synanamorph was associated with F. proteae and a new combination for P. proteae is proposed in Botryosphaeria, as B. proteae. The taxonomy of Botryosphaeria is in disarray at both the generic and the specific level. In Chapter 3 the taxonomic history of Botryosphaeria is reviewed, and the genus circumscribed and distinguished from other morphologically similar genera. Although several anamorph genera have been linked to Botryosphaeria, based on morphological observations and phylogenetic analysis of lTS rDNA sequence data, two anamorph genera are now recognised, those with pigmented conidia (Diplodia), and those with hyaline conidia (Fusicoccum). Botryosphaeria proteae should thus be excluded from Botryosphaeria. Several pathogenic Botryosphaeria spp. have an endophytic phase within their hosts. They are therefore imported unwittingly into other countries where they may pose a risk to agriculture and indigenous vegetation. The current global distribution of Botryosphaeria spp. associated with Proteaceae is clarified and a key to these taxa associated with Proteaceae is provided in Chapter 4. Five Botryosphaeria spp. are associated with cut-flower Proteaceae worldwide viz. B. lute a, B. obtusa, B. protearum, B. proteae and B. rib is. B. protearum is described as a new species. A thorough understanding of disease epidemiology is essential to effect a reduction of losses. In Chapter 5, I show that on P. magnifica, lesions caused by Botryosphaeria protearum, which lead to the formation of stem cankers, are initiated in the mid-rib vein or margin of leaves. Koch's postulates were satisfied and it was found that the number of lesions that developed from artificial inoculations correlated with starch levels present in leaves at the time of inoculation. In Chapter 6 it is shown that B. protearum exists as an endophyte in leaves of P. magnifica in naturally occurring as well as cultivated plants. In natural stands of proteas stem cankers are rare, but in cultivated plantations the incidence is high. Nutritional analyses indicate that higher levels of nitrogen occur in leaves of cultivated plants in spring, which could enhance disease development. High levels of sodium in the leaves of wild plants may restrict disease development. The severe economic losses caused by B. protearum make the search for improved methods of disease control essential. Fungicide applications form an important component of an integrated approach to disease management. In Chapter 7, in vitro tests demonstrate that tebuconazole, benomyl, prochloraz me, iprodione and fenarimol reduce the mycelial growth of B. protearum effectively. In the field there was a 25-85% reduction in the occurrence of stem cankers by applying fungicides or sanitation pruning. The best control was achieved by using benomyl, bitertanol, fenarimol, iprodione, prochloraz manganese chloride alternated with mancozeb and tebuconazole prophylactically. If sanitation pruning is combined with regular applications of fungicides, disease can be combated.
AFRIKAANSE OPSOMMING: Mikrofungi wat tot die genus Botryosphaeria behoort, is heterotrofiese organismes, wat patogenies op houtagtige plante kan wees. Hulle veroorsaak ernstige, en in sommige gevalle, verwoestende verliese, deur blaarnekrose, stamkankers en plantafsterwing. Die Proteaceae snyblom-industrie in Suid-Afrika maak 70% van die nasionale snyblomindustrie uit. Botryosphaeria siektes is 'n belangrike struikelblok in die produksie en handeldryf van Proteaceae, en daar is 'n ernstige behoefte om die etiologie, epidemiologie en beheer van siektes te ondersoek. Verliese van een van die belangrikste proteas, P. magnifica, beloop plaaslik 50% of meer. Die hoof doelstellings van hierdie studie was dus om die etiologie en epidemiologie van Botryosphaeria stamkankers op P. magnifica en ander Proteaceae vas te stel en metodes van siektebeheer te ondersoek. Hoewel daar 'n wye hoeveelheid inligting rakende die swam bestaan, wat in Hoofstuk I hersien is, is daar relatief min inligting oor Botryosphaeria op Proteaceae beskikbaar. Die taksonomie van Botryosphaeria benodig deeglike hersiening, en molekulêre tegnieke word benodig om spesie-identiteite op te klaar. In Hoofstuk 2 is gevind dat Phyllachora proteae, 'n blaarpatogeen van proteas, 'n Fusicoccum anamorf produseer, wat as F. proteae beskryf word. 'n Sphaeropsis-agtige synanamorf is met F. proteae geassosieer en 'n nuwe kombinasie vir P. proteae is as B. proteae in Botryosphaeria voorgestel. Die taksonomie van Botryosphaeria is, beide op die genus- as die spesievlak, in wanorde. In Hoofstuk 3 word die taksonomiese geskiedenis van Botryosphaeria hersien, en die genus word omskryf en van ander morfologies soortgelyke genera onderskei. Hoewel verskeie anamorf genera al met Botryosphaeria op grond van morfologiese waarnemings en filogenetiese analise van ITS rDNA volgorde data verbind is, word twee anamorf genera nou herken, dié met gepigmenteerde konidia (Diplodia), en dié met deurskynende konidia (Fusicoccum). Botryosphaeria proteae moet dus van Botryosphaeria uitgesluit word. Verskeie patogeniese Botryosphaeria spp. het 'n endofitiese fase in hul lewenssiklus. Hulle word dus onwetend in ander lande ingevoer waar hulle 'n gevaar vir landbou en inheemse plantegroei kan inhou. Die huidige wêreldverspreiding van Botryosphaeria spp. wat met Proteaceae geassosieer word is opgeklaar, en in Hoofstuk 4 word 'n sleutel tot die taksa wat met Proteaceae geassosieer word verskaf. Vyf Botryosphaeria spp. word met snyblom Proteaceae wêreldwyd geassosieer, naamlik B. lutea, B. protearum, B. proteae, B. ribis en B. obtusa. B. protearum word as 'n nuwe spesie beskryf. 'n Deeglike kennis van siekte-epidemiologie is noodsaaklik ten einde verliese te verminder. In Hoofstuk 5 dui ek aan dat letsels wat lei tot stamkankers, veroorsaak deur Botryosphaeria protearum op P. magnifica, in die hoofnerf of rant van blare ontstaan. Koch se postulate is uitgevoer en daar is vasgestel dat die aantal letsels wat vanuit kunsmatige inokulasies ontwikkel het korreleer met die styselvlakke teenwoordig in die blare ten tye van die inokulasie. In Hoofstuk 6 word getoon dat B. protearum as 'n endofiet in die blare van P. magnifica. In natuurlike standplase van proteas is stamkankers skaars, maar in verboude plantasies is die voorkoms hoog. Voedingsanalises dui aan dat hoër vlakke van stikstof in die blare van verboude plante in die lente voorkom, wat siekte-ontwikkeling moontlik kan bevorder. Hoë vlakke van natrium in die blare van natuurlike plante mag siekteontwikkeling beperk. Die ernstige ekonomiese verliese wat deur B. protearum veroorsaak word, maak die soektog na verbeterde metodes van siektebeheer noodsaaklik. Fungisiedtoedienings maak 'n belangrike deel uit van 'n geïntegreerde benadering tot siektebeheer. In Hoofstuk 7 dui in vitro toetse aan dat tebuconazole, benomyl, prochloraz me, iprodione en fenarimol die miseliumgroei van B. protearum effektief verminder. 'n Vermindering van 25-85% is aangetoon in die voorkoms van stamkankers in die veld, deur die toediening van fungisiedes en sanitasiesnoei. Die beste beheer is verkry deur die voorkomende toediening van benomyl, bitertanol, fenarimol, iprodione en prochloraz manganese chloride, afgewissel met mancozeb en tebuconazole, op plante in die veld. Indien sanitasiesnoei met gereelde toedienings van fungisiedes gekombineer word, kan die siekte bekamp word.

Thesis (MSc)--Stellenbosch University, 2004.
ENGLISH ABSTRACT: This thesis consists of four chapters that present research findings on Colletotrichum diseases associated with Proteaceae worldwide. The first chapter is a review of literature regarding the taxonomy and histology of Colletotrichum species associated with Proteaceae. The literature is not restricted to Proteaceae hosts, as information regarding Colletotrichum on Proteaceae is very limited. In chapter two, Colletotrichum spp. associated with proteaceous hosts growing in various parts of the world were identified based on morphology, sequence data of the internal transcribed spacer region (ITS-I, ITS-2), the 5.8S gene, and partial sequences of the B-tubulin gene. Four species of Colletotrichum were associated with Proteaceae. Colletotrichum gloeosporioides was isolated from Protea cynaroides cultivated in South Africa and Zimbabwe and from a Leucospermum sp. in Portugal, but is known to occur worldwide on numerous hosts. A recently described species, C. boninense, was associated with Zimbabwean and Australian Proteaceae, but also occurred on a Eucalyptus sp. in South Africa. This represents a major geographical and host extension for the species, and a description of the African strains is provided. Colletotrichum crassipes was represented by a single isolate obtained from a Dryandra plant in Madeira. Colletotrichum acutatum was isolated from Protea and Leucadendron in South Africa as well as from other proteaceous hosts occurring elsewhere. Colletotrichum acutatum f. sp. hakea was isolated from Hakea in South Africa. In chapter three, pathogenicity of these Colletotrichum species to certain proteas was established, relative aggressiveness of the different species tested and host response to them were compared as well as the effect that wounding had on host response. From the results obtained it is concluded that C. acutatum and C. gloeosporioides are the primary pathogens associated with Colletotrichum leaf necrosis, and C. acutatum is the main cause of anthracnose and stem necrosis of Proteaceae in South Africa. A histological study was performed in chapter four in response to the findings from the previous chapter. The behaviour of two C. acutatum isolates (one originating from Protea and the other from Hakea, C. acuataum f.sp. hakea) was studied on inoculated Protea leaf surfaces using light and scanning electron microscopy. Colletotrichum acutatum from Protea formed melanised appressoria on the leaf surface, whereas C. acutatum from Hakea formed very low numbers of both melanised and unmelanised appressoria. Most of the appressoria formed by C. acutatum from Protea were formed on the cell junctions and on the periclinal walls of the epidermal cells. From this study it is clear that C. acutatum f. sp. hakea is not a pathogen of Protea. Consequently the current use of this isolate as a biological control agent of Hakea in South Africa poses no threat to indigenous Protea species. Colletotrichum acutatum from Protea (although closely related to C. acutatum f. sp. hakea), is a pathogen of Protea, which was confirmed by histological observations. In conclusion, the present study has shown that several species of Colletotrichum are associated with diseased Proteaceae. These species differed in their pathogenicity and aggressiveness when inoculated onto certain protea cultivars. These differences could be partially explained by examining the behaviour of C. acutatum on the leaf surface. It is clear, however, that the distribution of the different species, their aggressrveness on different Proteaceae and their modes of infection needs to be investigated further. This work provides a basis for future research on the long-term effective management of these pathogens in fynbos production.
AFRIKAANSE OPSOMMING: Hierdie tesis bestaan uit vier hoofstukke wat handeloor navorsing van Colletotrichum siektes van Proteaceae wêreldwyd. Die eerste hoofstuk is 'n oorsig van literatuur rakende die taksonomie en histologie van die Colletotrichum spesies wat met Proteaceae geassosieer word. Die literatuur oorsig is nie beperk tot die Proteaceae nie aangesien baie min inligting rakende Colletotrichum op Proteaceae bestaan. In die tweede hoofstuk word die Colletotrichum spesies wat met proteas in verskeie dele van die wêreld geassosieer word, op grond van morfologie, DNS volgorde data van die interne getranskribeerde spasieerder area ("ITS-I, ITS-2"), die 5.8S geen, en gedeeltelike DNS volgordes van die B-tubulin geen geïdentifiseer. Vier Colletotrichum spesies is met die Proteaceae geassosieer. Colletotrichum gloeosporioides is geïsoleer vanaf Protea cynaroides wat in Suid-Afrika en Zimbabwe gekweek is en vanaf 'n Leucospermum sp. in Portugal, maar is bekend op verskeie gashere wêreldwyd. 'n Spesie wat onlangs beskryf is, C. boninense, is met Zimbabwiese en Australiaanse Proteaceae geassosieer, maar kom ook op 'n Eucalyptus sp. in Suid-Afrika voor. Dit is 'n groot uitbreiding van die geografiese voorkoms en gasheerreeks van hierdie spesie en 'n beskrywing van die Afrikaanse rasse word gegee. Colletotrichum crassipes is verteenwoordig deur 'n enkele isolaat wat vanaf 'n Dryandra plant in Madeira verkry is. Colletotrichum acutatum is vanaf Protea en Leucadendron in Suid-Afrika asook vanaf ander proteas wat elders voorkom, geïsoleer. Colletotrichum acutatum f. sp. hakea is vanaf Hakea in Suid-Afrika geïsoleer. In hoofstuk drie is die patogenisiteit van hierdie Colletotrichum spesies teenoor sekere proteas getoets, die relatiewe aggressiwiteit van die verskillende spesies is vergelyk, asook die gasheer se reaksie teenoor die spesies en die effek wat verwonding op die gasheer gehad het. Daar kan afgelei word vanaf die resultate dat C. acutatum en C. gloeosporioides die primêre patogene is wat met Colletotrichum blaarnekrose geassosieer word, en dat C. acutatum die hoof oorsaak is van antraknose en lootnekrose van Proteaceae in Suid-Afrika. 'n Histologiese studie is in hoofstuk vier uitgevoer in reaksie op die bevindings van die vorige hoofstuk. Die gedrag van twee C. acutatum isolate (een vanaf Protea en die ander vanaf Hakea, C. acutatum f.sp. hakea) op die oppervlaktes van geïnokuleerde Protea blare is bestudeer deur gebruik te maak van lig- en skandeer-elektronmikroskopie. Colletotrichum acutatum vanaf Protea vorm gemelaniseerde appressoria op die blaaroppervlak, terwyl C. acutatum vanaf Hakea klein hoeveelhede van beide gemelaniseerde en ongemelaniseerde appressoria vorm. Meeste van die appressoria wat deur C. acutatum vanaf Protea gevorm word, vorm op die aanhegtingspunte tussen selle en op die periklinale wande van die epidermale selle. Vanuit hierdie studie is dit duidelik dat C. acutatum f. sp. hakea nie 'n patogeen van Protea is nie. Gevolglik hou die huidige gebruik van hierdie isolaat as biologiese beheer agent van Hakea in Suid-Afrika geen gevaar in vir inheemse Protea spesies nie. Colletotrichum acutatum vanaf Protea (alhoewel dit naverwant is aan C. acutatum f. sp. hakea) is 'n patogeen van Protea en hierdie stelling is ook bevestig deur histologiese waarnemmgs. Ter samevatting het hierdie studie getoon dat verskeie Colletotrichum spesies geassosieer word met siektes van Proteaceae. Hierdie spesies het van mekaar verskil rakende patogenisiteit en aggressiwiteit nadat hulle op sekere protea kultivars geïnokuleer is. Hierdie verskille kon gedeeltelik verklaar word deur die gedrag van C. acutatum op die blaaroppervlaktes van verskillende protea kultivars. Dit is duidelik dat die verspreiding van die verskillende spesies, hulle aggressiwiteit op verskillende Proteaceae en hul infeksie metodes verder ondersoek moet word. Hierdie studie verskaf 'n basis vir toekomstige navorsing rakende lang-termyn effektiewe bestuur van hierdie patogene in fynbos aanplantings.

The effects of intraspecific and interspecific competition, host plant, and prior host plant experience on fecundity, rate of oviposition and mortality of adults and larval survival of Delia radicum Bouche (Cabbage Maggot (CM)), D. antiqua Meigen (Onion Maggot (OM)), and D. platura Meigen (Seed Corn Maggot (SCM)), were studied on cabbage, onion and bean plants.
An optimum density for maximum fecundity per female was observed when the four experimental densities were compared. This optimum density was higher on host than on non-host plants. CM females were host specific and did not oviposit on non-host plants. Rates of oviposition and mortality over a 30-day period were calculated for each density. The rate of oviposition was slower at higher densities on host plants for CM, OM and SCM. The rate of mortality increased at the highest density for CM (cabbage), OM (bean) and SCM (cabbage), but remained unaffected for OM on onion and cabbage and for SCM on onion and bean, when densities were compared. For OM, a delay in the rate of oviposition and mortality on cabbage (non-host plant) when compared with onion (host plant), suggests that cabbage was not as readily accepted as an oviposition site. Interspecific competition experiments at six density ratio's of SCM:OM indicated increased fecundity, or an increase in the rate of oviposition for OM, at the lower densities when single and mixed species were compared. For SCM no effects on fecundity were recorded, but the rate of oviposition was slower and rate of mortality faster at the lowest density in the presence of OM. Similar studies with SCM and CM showed no such effects of competition.
Host plant exposure of SCM females during the pre-oviposition period resulted in a delay in initial acceptance of subsequent host plants as oviposition sites. This happened only when females were exposed to a secondary host during the pre-oviposition period. Once oviposition began, host discrimination ceased and a switch in oviposition sites to the preferred host did not alter the rate of oviposition. In CM, the rate of larval development increased at density 6 (optimum density). Above this density a decrease in the rate of development and a significant reduction in pupal weight was observed. Time required for fly emergence was not affected by increasing larval densities.

The market life of many fruits and vegetables can be extended through storage at low temperatures. Chilling injury (CI) is a major postharvest storage problem for tropical commodities. Storing these products at temperatures below their critical temperature may result in severe physiological disorders known as CI symptoms. Mangoes (Mangifera indica. L) are susceptible to CI when stored below 12 °C. Visual CI symptoms include uneven ripening, surface pitting, discoloration, shriveling and scalding. Research has been conducted to overcome these serious problems using various postharvest treatments such as hot water, methyl jasmonate (MJ) or diphenylamine (DPA) with some reduction of the incidence of CI symptoms in fruits and vegetables.
Experiments were performed to assess and compare the potential of the above-mentioned postharvest treatments to reduce the CI symptoms on mango cv. Kent. The obtained results indicated that MJ- and DPA-treatments gave significantly greater percentage of marketable fruits.
Experiments were also conducted with mangoes cv. Tommy Atkins treated with MJ and DPA before storing at low temperatures (1, 4, 7 and 10°C). The chemical treatments were successful at reducing CI symptoms of mangoes. Fruit decay was reduced during subsequent ripening. MJ-treated fruits had lower mass loss and higher total soluble solids (TSS) than the control treatment. The overall quality of MJ- and DPA-treated fruits was good with lower surface pitting and scalding compared with the control treatment. The best results were obtained at storage temperatures of 7 and 10°C. Both MJ and DPA postharvest treatments can reduce CI symptoms in mangoes cvs. Kent and Tommy Atkins when the mangoes are stored at below critical temperature.

Thesis (PhD)--University of Stellenbosch, 2000.
ENGLISH ABSTRACT: The population density of the white wax scale, Ceroplastes destructor Newstead, has increased since 1994 in certain areas of Western and parts of Eastern Cape Provinces of South Africa where citrus is grown, particularly on Citrus reticulata (Blanco). A study was conducted to investigate its morphology, biology and ecology as contributions to the development of a sound integrated management programme. Characteristics of the immature stages and adult females were described and illustrated from field-collected and slide-mounted specimens. A key to the different stages and morphometeric characteristics useful for separating them are provided. No significant differences in female fecundity were found between orchards (P > 0.05). However, fecundity varied significantly between female size classes from the same orchard (P < 0.001). Female body-size also differed significantly between orchards (P < 0.05) and was significantly positively correlated with fecundity (P < 0.01). C. destructor has one discrete generation per year in South Africa. Oviposition commenced in November and continued through to the end of December with a few females ovipositing until mid January. Population density of the second instar peaked in February while the third instar extended from March to the end of July, followed by a peak population of adults in August. Seven primary and three secondary parasitoids, as well as four predator species attacking C. destructor were identified. Aprostocetus (= Tetrastichus) ceroplastae (Girault) was the dominant species, accounting for 78.87% of the total primary parasitoids reared. Peak numbers of parasitoids and predators were synchronized with peak emergence of susceptible scale stages, indicating that the host-parasitoid/predator system contained a density-dependent regulatory mechanism. Key mortality factors varied slightly between two of the orchards. Key stage mortality determined from a cohort life table was generally in the third instar (LIlI) and preovipositional female (POF) stage. Significant density-dependent mortality factors were demonstrated for the first instar (LI) and PDF stage. Dispersal of C. destructor is by first instar nymphs and the numbers caught on a series of yellow sticky traps varied significantly between crawler densities at the source, trap distances and trap directions from the source (P < 0.001). The numbers caught were positively correlated to the initial crawler density at the source (P < 0.01), suggesting that dispersal was density dependent. Trap distance and the numbers caught were inversely correlated (P < 0.01). Evaluation of effects of different densities of C. destructor on growth, survivorship and reproduction of scales as well as on leaf bearing ability of trees and area of leaf surface covered with sooty mould fungus was carried out on naturally infested Syzygium (= Eugenia) malaccensis (L.) plants. Scale body size and fecundity were inversely related to scale density (P < 0.01), suggesting density-dependent intraspecific competition. Scale survivorship generally declined with increasing density whereas scale parasitism and predation were positively correlated with density (P < 0.05). At high scale densities production of new leaves was significantly reduced (P < 0.01), reducing the resource base for subsequent generations of scale. Scale density and leaf area covered with sooty mould fungus were significantly positively correlated (P < 0.05). The toxicity of four synthetic insecticides against the three immature stages of C. destructor and of eight insecticides against the parasitoid A. ceroplastae was evaluated. Development of the first and second instars of C. destructor was completely arrested by the chemicals. Female fecundity, fertility and body sizes of survivors of treatments applied at the LIII stage were not significantly affected by any of the chemicals (P > 0.05). All the chemicals exhibited high toxicity to A. ceroplastae and hence are not recommended for integrated management of C. destructor in citrus orchards where A. ceroplastae plays an important role.
AFRIKAANSE OPSOMMING: Die populasiedigtheid van die witwasdopluis, Ceroplastes destructor Newstead, het sedert 1994 toegeneem in sekere gebiede van die Weskaap en Ooskaap provinsies van Suid-Afrika waar sitrus verbou word, veralop Citrus reticulata (Blanco). 'n Studie van hierdie insek se morfologie, biologie en ekologie is onderneem as bydrae tot die ontwikkeling van 'n geïntegreerde bestuursprogram. Die karaktertrekke van die onvolwasse stadia en die volwasse wyfies is beskryf en geïllustreer vanaf eksemplare wat in die veld versamel is en op g1asplaatjies gemonteer is. 'n Sleutel vir die verskillende stadia en morfometriese kenmerke wat nuttig is om hulle te onderskei, word voorsien. Geen beduidende verskille in die vrugbaarheid van wyfies van verskillende boorde is gevind nie (P < 0.05). Vrugbaarheid het egter betekenisvol verskil by die verskillende grootteklasse van wyfies uit dieselfde boord (P < 0.001). Die liggaamsgrootte van wyfies uit verskillende boorde het betekenisvol verskil (P < 0.05) en was betekenisvol positief gekorreleer met vrugbaarheid (P < 0.01). C. destructor het een generasie per jaar in Suid-Afrika. Eierlegging het in November begin en aangehou tot aan die einde van Desember, met enkele wyfies wat nog tot in middel Januarie eiers gelê het. Die populasiedigtheid van die tweede instar het 'n hoogtepunt in Februarie bereik, terwyl die derde instar van Maart tot aan die einde van Julie geduur het, gevolg deur 'n piekbevolking van volwassenes in Augustus. Sewe primêre en drie sekondêre parasitoïde asook vier predator spesies wat C destructor aanval, is geïdentifiseer. Aprostocetus (=Tetrastichus) ceroplastae (Girault) was die dominante spesies wat 78.87% van die totale aantal primêre parasitoïde wat uitgeteel is, uitgemaak het. Die pieke in die getalle van parasitoïde en predatore was gesinchroniseer met pieke in die verskyning van die gevoelige stadia, wat dui op die aanwesigheid van 'n digtheidsafhanklike regulatoriese meganisme. Die sleutel mortaliteitsfaktore het effens gevarieer tussen twee van die boorde. Die sleutelstadium van mortaliteit, soos bepaal m.b.v. 'n kohort lewenstabel, was gewoonlik die derde instar (LIlI) en die preoviposisionele wyfie (POW). Betekenisvolle digtheidsafhanklike mortaliteitsfaktore IS aangetoon vir die eerste instar (LI) en die POW. Die verspreiding van C.destructor vind plaas deur die eerste instar nimfe en die getalle wat op 'n reeks van taai geel valle gevang is, het betekenisvol gewissel volgens kruiperdigthede by die bron, asook die afstand en rigting van die valle vanaf die bron (P < 0.001). Die getalle wat gevang is, was positief gekorreleer met die aanvanklike kruiperdigtheid by die bron (P < 0.01), wat daarop dui dat verspreiding digtheidsafhanklik was. Die afstand van die valle en die aantal wat gevang is, was omgekeerd gekorreleer (P < 0.01). 'n Evaluering van die invloed van verskillende digthede van C. destructor op die groei, oorlewing en reproduksie van dopluise, asook die vermoë van bome om blare te dra en die area van die blaaroppervlak wat met roetskimmel besmet is, is uitgevoer op plante van Syzygium (= Eugenia) malaccensis (L.) met 'n natuurlike besmetting. Die liggaamsgrootte en vrugbaarheid van die dopluise was omgekeerd gekorreleer met hulle digtheid (P < 0.01), wat dui op digtheidsafhanklike intraspesifieke kompetisie. Die oorlewing van die dopluise het oor die algemeen afgeneem met toenemende digtheid, terwyl parasitisme en predasie positief gekorreleer was met digtheid (P < 0.05). By hoë dopluisdigthede het die produksie van nuwe blare betekenisvol afgeneem (P < 0.01), wat die hulpbronbasis vir daaropvolgende generasies van dopluise verswak. Die dopluisdigtheid en blaaroppervlak wat met roetskimmel bedek was, was positief gekorreleer (P < 0.05). Die toksisiteit van vier sintetiese insektemiddels teenoor die drie onvolwasse stadia van C. destructor en van agt insektemiddels teenoor die parasitoïd A. ceroplastae is geëvalueer. Die ontwikkeling van die eerste en tweede instars van C. destructor is heeltemal stopgesit deur die middels. Die fekunditeit, fertiliteit en liggaamsgrootte van wyfies wat toedienings op die LIIl stadium oorleef het, is nie betekenisvol ge-affekteer deur enige van die middels nie (P < 0.05). Al die middels was baie toksies teenoor A. ceroplastae en word dus nie aanbeveel vir die geïntegreerde bestuur van C. destructor waar A. ceroplastae 'n belangrike rol speel nie.

A new disease affecting potatoes was first detected in Mexico in 1993. Affected plants had aerial symptoms similar to those caused by potato purple top and psyllid yellows, but tubers had internal brown discoloration when sliced and dark stripes and streaks when processed to produce potato chips. The disease has been found in many potato production areas in Guatemala, Mexico, Honduras, New Zealand and the United States. The disease, termed Zebra Chip (ZC), has been associated with the presence of heavy infestations of the potato-tomato psyllid (Bactericera cockerelli). In 2009, a research group in New Zealand discovered that a new disease in tomato and pepper plants was caused by Candidatus Liberibacter solanacearum (Lso) and subsequently this same bacterium was associated with ZC in potato samples from Texas. The objectives of this study were: to assess the accumulation of Lso in various potato organs, to determine the effect of plant age on detection of Lso, symptom development and plant death, and (iii) to determine the effect of phosphorous acid on the development of ZC. Results from these studies showed significant differences in Lso populations between above and below ground tissues of the potato plant, with Lso populations in stolons and tubers being three to four times higher than those of leaf tissue and over seventy times greater than in stems. Time for detection of Lso by PCR in potato leaves of different ages at the time of inoculation ranged from 21 to 26 days after inoculation, symptoms development took 23 to 36 days. Plant death, took 24 to 47 days in plants of different age groups at the time of inoculation. In plants 15 weeks old at the time of inoculation, Lso was detected after 14 days in one plant out of 18; in plants 16 weeks old at the time of inoculation, Lso was detected after seven days in two plants out of 18. Phosphorous acid applications had no effect on the populations of Lso in potato tubers, onset of symptoms or plant death. All tubers showed ZC symptoms, making them unacceptable for the market.
North Dakota State University. Department of Plant Pathology

Volatile metabolites from headspace gas of mango cultivars Tommy Atkins and Keitt, wounded and inoculated with two pathogens, Colletotrichum gloeosporioides and Lasiodiplodia theobromae or non-inoculated controls were profiled using a GC/MS to develop a technology to discriminate diseases. Several disease discriminatory compounds were identified and classified into three groups: (i) compounds unique to only one treatment; (ii) compounds common to two or more treatments but not to all; and (iii) compounds common to all treatments but with varying in their abundance. 1-pentanol and boronic acid ethyl were detected in only Lasiodiplodia-inoculated mangoes while thujol was observed only in Colletotrichum-inoculated mangoes. Models based on significant mass ions classified up to 100% of the diseases/inoculations. The disease discriminatory compounds and discriminant analysis models developed here could be used in the early detection of postharvest diseases of mango fruit, after validation under commercial conditions.

Dissertation (PhD (Agric))--University of Stellenbosch, 2003.
ENGLISH ABSTRACT: Leaf blackening is a particular problem limiting vase life and marketability of Protea cut flowers. This research investigated suppression of Protea leaf blackening with a specific focus on Protea cv. Sylvia (P. eximia x P. susannae) cut flowers. Leaf blackening decreased significantly with decreasing storage temperatures m 'Sylvia' proteas and this was attributed to lower respiration rate and conservation of carbohydrate. Low storage temperatures were beneficial in short term handling procedures encountered during airfreight. However, use of low temperatures alone during the longer sea freight period was unsatisfactory in either maintaining or extending 'Sylvia' protea vase life. Cooling of 'Sylvia' proteas under vacuum significantly suppressed leaf blackening and was of greater benefit than forced air cooling. Although removal of the uppermost leaves delayed leaf blackening in short term storage no significant benefit was found for longer storage periods. Girdling directly beneath the 'Sylvia' protea flowerhead significantly reduced leaf blackening and in combination with low storage temperatures (O°C) enabled a significant extension in both storage and vase life of 'Sylvia' proteas. 'Sylvia' proteas did not exhibit a climacteric respiration peak during 96 h storage at O°C. Exposure to ethylene did not increase Protea leaf blackening or have a detrimental effect on vase life of either proteas or pincushions evaluated. No beneficial response to sucrose supplementation was found in 'Sylvia' proteas. Analysis of the sugar content of both flowerhead and leaves indicated that glucose supplementation might be of benefit and was investigated. Holding solutions of 2.5 % glucose significantly extended vase life due to a significant reduction in leaf blackening. Vase life was terminated due to flowerhead collapse instead of leaf blackening for the first time in 'Sylvia' protea cut flowers. Vase life was significantly extended by 2:3% glucose pulse solutions and leaf blackening significantly suppressed with increasing glucose pulse concentration. Solution uptake was facilitated by use of high intensity PAR lights in the early morning and was attributed to increased stomata opening and a consequent increase in both transpiration and glucose solution uptake. The faster uptake of glucose solutions in shoots harvested in the afternoon was attributed to higher shoot temperatures and consequent transpiration rate to those harvested in the morning. There was a significant reduction in uptake time with increasing pulse temperature, which enabled vacuum cooling to be performed earlier further benefiting storage and vase life extension. Enclosure of 'Sylvia' proteas in polyethylene (PE) lined cartons did suppress leaf blackening in non-pulsed shoots. However, this had no practical significance on useful vase life, which was terminated at this point due to excessive leaf blackening. Water loss appears to have a minimal influence on 'Sylvia' protea leaf blackening. Shading at four and three weeks prior to harvest coincided with a period of significant flowerhead dry mass increase. It is thought that shading at this point, concurrent with an increased carbohydrate demand by the developing flower head resulted in a temporary limitation in carbohydrate supply resulting in the appearance of preharvest leaf blackening. It would appear that proteas do not store large quantities of carbohydrate. Although accentuating winter light conditions by shading did result in a decrease in carbohydrate content the fact that carbohydrate content was already low precluded shading from having a significant impact on postharvest leaf blackening. The finding that glucose was beneficial in extension of both storage and vase life of 'Sylvia' proteas directed research into its use for other Protea and Leucospermum cut flowers. Significant differences in the response to glucose supplementation were found in both Protea and Leucospermum (pincushions). The significant difference in sensitivity to glucose concentration in 'Pink Ice' proteas (phytotoxic at 2:4%) and 'Susara' proteas (no apparent toxicity), in conjunction with a lack of response in 'Cardinal' proteas, a hybrid from the same parents as 'Sylvia' indicates the need to direct future research to individual cultivars. Glucose supplementation had no beneficial effect on vase life of 'Scarlet Ribbon' and 'Tango' pincushions, whilst significantly extending vase life of 'Cordi', 'Gold Dust', 'High Gold' and 'Succession' pincushions.
AFRIKAANSE OPSOMMING: Blaarverswarting is 'n spesifieke probleem wat die vaasleeftyd en die bemarkbaarheid van Protea snyblomme beperk. In hierdie navorsing is ondersoek ingestel na die onderdrukking van Protea blaarverswarting met spesifieke fokus op die snyblomme van die kv. Sylvia (P. eximia x P. susannae). Die voorkoms van blaarverswarting by 'Sylvia' het merkbaar afgeneem tydens die verlaging van bergingstemperature. Hierdie afname is toegeskryf aan 'n laer respirasietempo en die behoud van koolhidrate. Lae bergingstemperature in die korttermyn hantering van die produk tydens lugvrag was voordelig. Die gebruik van lae temperature, slegs tydens die langer verskeepingsperiode, was egter onbevredigend vir vaasleeftyd verlenging en onderhoud van 'Sylvia' protea. Die afkoeling van 'Sylvia' proteas onder vakuum het blaarverswarting in 'n groot mate onderdruk en het beter resultate gelewer as geforseerde lugverkoeling. Alhoewel die verwydering van die heel boonste blare blaarverswarting by korttermynopberging vertraag het, het dit geen merkbare voordele vir langer bergingsperiodes ingehou nie. Ringelering direk onder die blomkop van die 'Sylvia' protea het blaarverswarting aansienlik verminder, en saam met lae bergingstemperature (O°C) het dit 'n merkbare verlenging in beide die bergingstyd en die vaasleeftyd van 'Sylvia' proteas teweeggebring. 'Sylvia' proteas het geen klimakteriese respirasiekruin tydens 'n bergingsperiode van 96 uur teen O°C getoon nie. Blootstelling aan etileen het nie die Protea blaarverswarting laat toeneem of 'n nadelige effek op die vaasleeftyd van die proteas of speldekussings wat geevalueer is, gehad nie. Geen voordelige reaksie op sukrose-byvoeging is in 'Sylvia' proteas gevind nie. 'n Analise van die suikerinhoud van beide die blomkoppe en die blare het aangetoon dat 'n glukose-byvoeging moontlik voordelig kon wees, en hierdie aspek is ondersoek. Met stooroplossings van 2,5 % glukose is die vaasleeftyd aansienlik verleng omdat daar 'n merkbare afname in blaarverswarting was. Vir die eerste keer in die geval van die 'Sylvia' protea, het die vaasleeftyd van die snyblomrne tot 'n einde gekom omdat die blornkoppe uitmekaar gebreek het en nie omdat blaarverswarting ingetree het nie. Die vaasleeftyd is aansienlik verleng met ~ 3% glukose-pulsoplossings, en blaarverswarting is merkbaar onderdruk met die verhoging van hierdie oplossings se glukosekonsentrasie. Die opname van die oplossings is gefasiliteer deur hoe intensiteit PAR (fotosinteties-aktiewe radiasie) ligte vroeg in die oggend, en is toegeskryf daaraan dat meer huidmondjies oopgegaan het. Dit het gelei tot 'n toename in transpirasie en 'n toename in die opname van die glukose-oplossing. Die feit dat glukose-oplossings vinniger opgeneem is deur lote wat in die middag geoes is, is toegeskryf daaraan dat loottemperature dan hoer is as soggens en gevolglik lei tot 'n vinniger transpirasietempo. Daar was 'n merkbare afname in die opnametyd wanneer die temperatuur van die pulsoplossings verhoog is. Vakuumafkoeling kon dus vroeer toegepas word, wat 'n verlenging in bergingstyd en vaasleeftyd tot gevolg gehad het. Verpakking van 'Sylvia' proteas in kartonne wat met poli-etileen uitgevoer is, het blaarverswarting van lote wat nie aan pulsering onderwerp is nie, onderdruk. Hierdie maatreel het egter geen praktiese waarde met betrekking tot vaasleeftyd nie; die vaasleeftyd het tot 'n einde gekom as gevolg van omvangryke blaarverswarting. Dit lyk asof waterverlies weinig invloed het op die blaarverswarting van' Sylvia' proteas. Die vermoede bestaan dat lae koolhidraatvlakke proteas ontvanklik maak vir blaarverswarting. Alhoewel die beklemtoning van winterligtoestande deur beskaduwing gelei het tot 'n afname in koolhidraatinhoud, was hierdie inhoud reeds laag en blaarverswarting na die oes is nie beinvloed nie. Beskaduwing tydens die vier en drie weke voor oestyd het saamgeval met 'n tydperk van aansienlike toename in die droe massa van die blomkop. Die vermoede bestaan dat beskaduwing tydens hierdie fase, saam met die toename in die ontwikkelende blomkop se behoefte aan koolhidrate, aanleiding gegee het tot 'n tydelike beperking in koolhidraatvoorraad wat die voorkoms van blaarverswarting voor die oes tot gevolg gehad het. Die bevinding dat glukose voordelig is vir die verlenging van beide die bergingstyd en die vaasleeftyd van 'Sylvia' proteas het die navorsing gerig om ook ondersoek in te stel na die gebruik daarvan vir ander Protea en Leucospermum snyblomme. Merkbare veranderinge is gevind in die reaksie op glukosebyvoegings in beide Protea en Leucospermum (speldekussings). Die opmerklike verskil in sensitiwiteit vir glukosekonsentrasie in 'Pink Ice' proteas (fitotoksies by ~ 4%) en 'Susara' proteas (geen klaarblyklike toksisiteit), saam met 'n gebrek aan reaksie by 'Cardinal' proteas, 'n hibried van dieselfde ouers as 'Sylvia', dui aan dat verdere navorsing op individuele kultivars toegespits sal rnoet word. Glukosebyvoegings het geen voordelige uitwerking op die vaasleeftyd van 'Scarlet Ribbon' en 'Tango' speldekussings gehad nie, terwyl dit die vaasleeftyd van 'Cordi', 'Gold Dust', 'High Gold' en 'Succession' speldekussingkultivars merkbaar verIeng het.

Soybean (Glycine max [L.] Merr.) is one of the most important crops in the world. The average annual yield losses due to soybean diseases and pests are estimated to be around 11% in the United States. Soybean yield losses due to sudden death syndrome (SDS), caused by the fungus Fusarium virguliforme O'Donnell & T. Aoki have been problematic in majority of the soybean producing states. In recent years, reniform nematode (RN, Rotylenchulus reniformis Linford and Oliveira) and frogeye leaf spot (FLS), caused by the fungus Cercospora sojina K. Hara have emerged as a major problem in the southern soybean producing states. Planting resistant cultivars is one of the most cost-efficient methods in managing SDS, RN, and FLS, therefore it would be critical to identify and map the quantitative trait loci (QTL) that underlie their resistances. Two soybean populations were evaluated in this study. The ‘Essex’ × ‘Forrest’ 77 near-isogenic lines were screened in the field to evaluate the disease index of SDS. The Essex × Forrest and ‘Flyer’ × ‘Hartwig’ recombinant inbred lines were screened in the greenhouse to assess the reproduction index of RN and the disease severity of FLS. The BARCSoySNP6k chip was used to genotype the two populations. Four QTL that underlie SDS resistances were mapped in the same region as Rfv06-01, Rfv06-02, Rfv13-01, and Rfv19-01. The Rfv06-02 interval in this study was smaller than the one previously reported. Rrr08-01, Rrr13-01, Rrr15-01, Rrr18-01, and Rrr18-02 were reported to confer resistances to RN. Rrr08-01, Rrr13-01 and Rrr15-01 were novel whereas Rrr18-01, and Rrr18-02 were mapped in previous studies. cqSCN-001 (soybean cyst nematode, Heterodera glycines Ichinohe) was identified in the same region as Rrr18-01, and Rrr18-02 whereas cqSCN-006 was identified in the same region as Rrr15-01. These findings provide further evidence that there are common sources of genetic resistances to RN and SCN. Rcs15-01 and Rcs15-02 were reported to confer resistances to FLS. Rcs15-01 was novel and Rcs15-02 was mapped at the same region as an Rcs mapped in a previous study. This indicated that Rcs15-02 has dual resistances to C. sojina races. Candidate genes were inferred in this study. The QTL mapped in this study could potentially be used in soybean breeding programs that aim to introgress genetic resources that confer resistances to SDS, RN, and FLS.

In recent years the hot water treatment (HW) represents an effective and safe approach for managing postharvest decay. This study reported the effect of an HW (60°C for 60 s and 45°C for 10 min) on brown rot and blue mould respectively. Peaches was found more thermotolerant compared to apple fruit, otherwise Penicillium expansum was more resistant to heat with respect to Monilinia spp. In semi-commercial and commercial trials, the inhibition of brown rot in naturally infected peaches was higher than 78% after 6 days at 0°C and 3 days at 20°C. Moreover, in laboratory trials a 100% disease incidence reduction was obtained by treating artificially infected peaches at 6-12 h after inoculation revealing a curative effect of HW. The expression levels of some genes were evaluated by qRT-PCR. Specifically, the cell wall genes (β-GAL, PL, PG, PME) showed a general decrease of expression level whereas PAL, CHI, HSP70 and ROS-scavenging genes were induced in treated peaches compared to the control ones. Contrarily, HW applied on artificially infected fruit before the inoculum was found to increase brown rot susceptibility. This aspect might be due to an increase of fruit VOCs emission as revealed by PTR-ToF-MS analysis. In addition a microarray experiment was conducted to analyze molecular mechanisms underneath the apple response to heat. Our results showed a largest amount of induced Heat shock proteins (HSPs), Heat shock cognate proteins (HSCs), Heat shock transcription factors (HSTFs) genes found at 1 and 4 hours from the treatment. Those genes required for the thermotolerance process could be involved in induced resistance response. The hypothesis was confirmed by 30% of blue mold disease reduction in artificially inoculated apple after 1 and 4 hours from the treatment. In order to improve peaches quality and disease management during storage, an innovative tool was also used: Da-meter.

The influence of uniformity of heat transfer with respect to the effect of heat treatment was investigated by correlating engineering parameters with the quantified heat treatment effects. Differences in treatment effect in terms of quality, anti-pathogen and chilling injury (CI) control properties among commodities was studied by exposing them to different target temperatures. Similar effects were also evaluated and quantified within an individual commodity by a custom-designed device. This device, whose design parameters were based on a heat transfer simulation, insured that one hemisphere of a tomato was exposed to air at 39°C and 0.24 m s-1; while the other hemisphere was exposed to air at a lower temperature with a velocity of0.24 m s-1 or 0.12 m s-1.
Single-temperature heat treatment was most effective in limiting pathogen development and varied according to the parameter measured: 38°C for hypersensitive response (HR), 36°C for tissue breakdown, 36°C, 38°C or 39°C for mycelium abundance, and 38°C or 39°C for lesion size. Bilateral differences in temperature across the fruit significantly affected disease control: decreasing temperature differences significantly improved the uniformity of disease control.
Some of the effects of heat treatment on tomato quality, such as color development and resistance to CI, appear to be localized. A significant difference in redness was identified between heated parts and unheated parts of tomato fruits immediately after treatment; and the differences persisted during storage. Differences in lightness and chroma were noted on day 4. Delay in ripening caused by heat treatment was confirmed through the higher TA and TSS values of heated tomatoes or heated portion of partially heated tomatoes.
The heated parts of tomatoes showed a stronger resistance to chilling injury. The effective temperature control range for CI was wide, but temperatures higher than 39.5°C for 23 h hot air treatment could lead to adverse effects.
Differences in physiological effects between hemispheres in two chambers was reduced by directly decreasing the temperature difference between upper and lower chambers or relatively increasing the heating air flow rate, highlighting the importance of improving the uniformity of air flow around each individual treated fruit.

Bibliography: pages 175-184.
The occurrence of maize dwarf mosaic virus (MDMV) in field grown maize was investigated. For this purpose, maize showing mosiac symptoms was collected from different maize growing areas in South Africa by Prof. M.B. von Wechmar. These samples from Transvaal, Orange Free State and Natal were then investigated for the presence of MDMV and possible strains of this virus. Three virus isolates were purified and partially characterised. These isolates were serologically compared together with a fourth isolate SCMV 4975, obtained from the U.S., to establish strain relationships.

Thesis (PhD(Agric))--University of Stellenbosch, 2001.
ENGLISH ABSTRACT: Postharvest decay of stone fruit in the Western Cape province of South Africa is caused primarily by Botrytis cinerea (grey mould) and Monilinia laxa (brown rot). Little is known about the relative importance and seasonal occurrence of the two pathogens in nectarine and plum orchards, the mode of penetration of fruits by M laxa, latency and subsequent disease expression by the latter pathogen. These aspects were investigated in this study. By sampling from the Unifruco Quality Evaluation Scheme and from 11 stone fruit orchards, observations were made over a 3-year period of the occurrence of grey mould and brown rot in the major stone fruit regions. Botrytis cinerea was found to be the most important pathogen causing blossom blight and postharvest decay on stone fruit. The pathogen was most prominent on early- and mid-season culti~ars. Brown rot was exclusively caused by M laxa and no evidence was found that M fructicoZa had been introduced into the region. Monilina laxa was most prominent on the later maturing cultivars. Botrytis cinerea blossom infection did not contribute directly to postharvest decay. Both surface inoculum and latent infection consistently occurred on fruit in each orchard, although at fluctuating levels. Disease expression on developing fruit was not governed by the amount of B. cinerea occurring on fruit surfaces, but by the ability of fruit to resist disease expression. The amount of B. cinerea on fruits was generally higher during spring than during summer. Monilinia laxa occurred sporadically on the blossoms of late-maturing cultivars. Immature fruit were generally pathogen-free and disease expression occurred on maturing fruit only. These findings suggest that conidia of M laxa are generally produced in orchards when fruits are approaching maturity and can penetrate and infect maturing fruit only. The behaviour of airborne M laxa conidia was subsequently studied on nectarine (cultivar Flamekist) and plum (cultivar Laetitia) fruit. For these studies, an inoculation method that simulates natural infection by airborne conidia was used. Fruit at pit hardening, 2 wk before harvest, harvest stage and after cold storage (nectarines 4 wk at -o.soC followed by 1 wk at 23°C at ±56% RH; plums 10 days at .....().5°C,18 days at 7.5°C followed by 1 wk at 23°C at ±56% RH) were dusted with dry conidia of M laxa in a settling tower. The fruits were incubated for periods ranging from 3 to 48 h at high relative humidity (2':93%, humid fruit) or covered with a film of water (wet fruit). Behaviour of the solitary conidia was examined with an epifluorescence microscope on skin segments stained in a differential stain containing fluorescein diacetate, aniline blue and blankophor. The ability of solitary conidia to colonise the fruit surface, penetrate fruit skins and to induce disease expression was determined by using a differential set of tests. For these tests, fruit were surface-sterilised (30 s in 70% ethanol) or left Unsterile. From each group, fruit were selected for isolation (skin segment test), immersed in a 3% paraquat solution (paraquat-treated fruit test) or left untreated (sound fruit test). 1be findings demonstrated that solitary conidia of M laxa behaved consistently on plum and nectarine fruit surfaces: appressorium formation and direct penetration was not observed on any of the fruit surfaces and germ tubes penetrated fruit predominantly through stomata, lenticels and microfissures in the fruit skin. The monitoring of airborne conidia revealed subtle effects of the fruits on the behaviour of solitary germlings, which could not be seen when using conidial suspensions. On both fruit types, no deleterious effect was seen on conidial and germling survival when fruit were kept humid at pit hardening, 2 wk before harvest and harvest. However, conidial and germling survival were drastically reduced by prolonged wet incubation of fruits. The findings on disease expression in the skin segment, paraquat-treated fruit and sound fruit tests clearly showed that the skin of both nectarine and plum fruits were not penetrated at the pit hardening stage, latent infections were not established and fruitsreacted resistant to disease expression. These facets on both fruit types were furthermore unaffected by wetness. The barrier capacity of the fruit skin of the two stone fruit types however differed drastically later in the season. On nectarine, fruit skins were more readily penetrated and disease expression became more pronounced when fruit approached maturity. Penetration and disease expression on ripening nectarine fruit were furthermore greatly influenced by wetness. Maturing plum fruit, on the other hand, did not display the drastic change in the barrier capacity of fruit skins as observed on nectarine. The influence of wetness on infection and disease expression was also less pronounced than on nectarine. In fact, plum fruit remained asymptomatic in the sound fruit test after inoculation and humid incubation at the 2 wk before harvest stage, harvest stage and after cold storage. Plum fruit at these stages only developed disease after a prolonged period (~12 h) of wet incubation. The paraquat fruit test revealed that these fruits became more susceptible to latent infection, but they were not as susceptible as nectarine. Collectively, these findings indicate that M. laxa fruit rot epidemics on plum and nectarine are driven by inoculum levels on fruit approaching maturity and by weather conditions prevailing during the preharvest and harvest period. However, the barrier capacity of plum skins is considerably more effective than that of nectarine fruit. Wounds would therefore play an important role in the epidemiology of M. laxa on plum fruit. Infection of fresh wounds by airborne M. laxa conidia, and by conidia and germlings that have established on fruits, was therefore investigated. Plum fruit (cultivar Laetitia) at pit hardening, 2 wk before harvest, harvest stage and after cold storage were dusted with dry conidia of M. laxa in a settling tower.- Infection of rionwounded fruit and of fresh wounds by \ the airborne conidia on dry, humid and wet plum fruit surfaces, and by conidia and germlings that have been established on fruits under the wetness regimes was then investigated. Nonwounded immature and mature fruit remained mostly asymptomatic, whereas nonwounded cold stored fruit decayed readily. Wounding drastically increased infection by airborne conidia. Immature fruits were less susceptible to wound infection by the airborne conidia than mature fruits. Conidia dispersed freshly were more successful in infecting fresh wounds than conidia that were deposited, or germlings that established, on fruit surfaces 4 days prior to wounding. This decrease in infectivity was especially pronounced on humid and even more on wet incubated fruit. This study clearly showed that in order to reduce. the incidence of brown rot, inoculum levels on fruit approaching maturity should be reduced by sanitation practices and fungicide applications. Furthermore, it is essential to protect fruits, especially. near-mature fruits, from being wounded.
AFRIKAANSE OPSOMMING: EPIDEMIOLOGIE VAN MONILINIA LAXA OP NEKTARIEN EN PRUIM: INFEKSIE VAN VRUGTE DEUR KONIDIA OPSOMMING Naoesverrotting van steenvrugte in die Wes-Kaap provinsie van Suid-Afrika word hoofsaaklik veroorsaak deur Botrytis cinerea (vaalvrot) en Monilinia laxa (bruinvrot). Min is bekend oor die relatiewe belang en seisoenale voorkoms van hierdie patogene in nektarienen pruimboorde, asook oor die infeksieweg, latensie en daaropvolgende siekte-uitdrukking van M laxa. Hierdie aspekte is in dié studie nagevors. \ \ Monsters IS oor 'n 3-jaar periode van die Unifruco Kwaliteitsevalueringskema, en ook van 11 steenvrugboorde verkry. Die voorkoms van vaalvrot en bruinvrot in die hoof steenvrugareas is so bepaal. Botrytis cinerea was die belangrikste patogeen wat betref bloeiselversenging en naoesverrotting. Verder was hierdie patogeen ook meer prominent op die vroeë- en middel-seisoen kultivars. Bruinvrot is uitsluitlik deur M Iaxa veroorsaak en geen aanduiding omtrent die moontlike voorkoms van M fructicola in Suid-Afrika is waargeneem nie. Monilinia laxa was meer prominent op die laat-seisoen kultivars. Botrytis cinerea bloeiselinfeksie het nie direk bygedra tot naoesverrotting nie. Beide oppervlakkige inokulum en latente infeksie het deurgaans, maar wel teen wisselende hoeveelhede, op vrugte in die onderskeie boorde voorgekom. Siekte-uitdrukking op ontwikkelende vrugte is egter nie beinvloed deur die hoeveelheid B. cinerea op die vrug nie, maar eerder deur die vermoë van die vrug om siekte-uitdrukking te onderdruk. Die hoeveelheid B. cinerea op vrugte was verder hoër gedurende lente as gedurende somer. Monilinia laxa het slegs sporadies op die bloeisels van laat-seisoen kultivars voorgekom. Groen vrugte was in die algemeen vry van die patogeen en siekte-uitdrukking het slegs op ryp vrugte plaasgevind. Hierdie bevindinge dui daarop dat M laxa in boorde hoofsaaklik op ryper vrugte geproduseer word. Hierdie swam infekteer ook net ryp vrugte. Die gedrag van luggedraagde M laxa conidia is bestudeer op nektarien- (kultivar Flamekist) en pruimvrugte (kultivar Laetitia). 'n Inokulasie-metode wat natuurlike infeksie deur luggedraagde konidia simuleer, is vir hierdie studies gebruik. Vrugte van die pitverharding-, twee weke voor oes-, oesstadium, asook koud-opgebergde vrugte (nektariene, 4 weke by -o.soe gevolg met 1 week by 23°C en ±56% RH; pruime, 10 dae by -O.5°e, 18 dae by 7.Soe gevolg deur 1 week by 23°C en ±56% RH), is met droë konidia in 'n inokulasietoring geïnokuleer. Die vrugte is vir periodes wat gewissel het van 3 tot 48 h geïnkubeer by hoë relatiewe humiditeit (~93% RH, vogtige vrugte), of dit is bedek met'n film water (nat vrugte). Die gedrag van die enkelspore (konidia) op die vrugoppervlak is met 'n epifluorisensiemikroskoop bestudeer. Skilsegmente is gekleur in 'n kleurstof, bevattende fluorisein diasetaat, analien-blou en blankofor. Die vermoë van die enkelspore om die vrugoppervlak te koloniseer, te penetreer en om siekte-uitdrukking te induseer, is met 'n differensiële stel toetse bepaal. Vir hierdie toetse is die vrugte oppervlakkig gesteriliseer (30 s in 70% etanol), of nie-steriel gelaat. In elke groep is vrugte geneem vir isolasie (skilsegment-to\~ts), of gedoop in "n 3% parakwat-oplossing (parakwat vrugtoets), of\, onbehandeld gelaat (onbehandelde vrugtoets ). Die. bevindinge het op die soortgelyke gedrag van M laxa enkelspore op die verskillende vrugsoorte gedui: appressoria en direkte penetrasie is nie waargeneem nie, en kiembuise het die vrugte hoofsaaklik deur huidmondjies, lentiselle en mikro-krakies .in die vrugskil gepenetreer. Deur luggedraagde spore te bestudeer, is sekere subtiele effekte van die vrug op die gedrag van enkelspore op die vrugoppervlak waargeneem. Op beide vrugtipes is geen nadelige effek op konidiurn- en kiembuisoorlewing opgemerk wanneer die vrugte onder hoë vogtoestande geïnkubeer is. Konidiurn- en kiembuisoorlewing is egter drasties verlaag hoe langer die vrugte onder nat toestande geïnkubeer is. Die bevindinge van die skilsegment-, parakwat en onbehandelde vrugtoetse het duidelik daarop gewys dat die vrugskil van nektarien en pruim nie gepenetreer is tydens die pitverhardingstadium nie, latente infeksies is nie gevorm nie, en die vrugte was bestand teen siekte-uitdrukking. Hierdie fasette op beide vrugtipes is ook nie beinvloed deur inkubasie-natheid nie. Die beskermingskapasiteit van die vrugskil van hierdie steenvrugtipes het egter drasties verskil later in die seisoen. Nektarien-vrugskille is meer geredelik gepenetreer en siekte-uitdrukking het toegeneem met rypwording. Penetrasie en siekteuitdrukking is verder in 'n groot mate deur inkubasie-natheid bevoordeel. Rypwordende pruime het egter nie so In drasties verandering in die beskermingskapasiteit van die vrugskil getoon nie. Die invloed van inkubasie-natheid op infeksie en siekte-uitdrukking was ook minder opsigtelik as op nektarien. Pruimvrugte van die twee weke voor oes-, oesstadium, en , koud-opgebergde pruime, wat onder hoë vog geïnkubeer is, het simptoomloos in die onbehandelde vrugtoets gebly. Vrugte van hierdie stadia het slegs simptome ontwikkel na periodes van langer as 12 h onder nat toestande. Die parakwat-behandelde vrugtoets het egter gewys dat die pruimvrugte meer vatbaar vir latente infeksies raak, maar steeds nie so vatbaar soos die nektarienvrugte nie. Gesamentlik dui hierdie bevindinge daarop <41tM laxa bruinvrot epidemies op pruim en nektarien afhanklik is van inokulumvlakke op rypwordende vrugte, asook die weerstoestande gedurende die vooroes- en oesstadia. Die beskermingskapasiteit van pruim vrugskille was egter aansienlik meer effektief as dié van nektarien vrugte. Wonde op vrugte sal dus 'n groter rol speel in die epidemiologic van M laxa op pruim. Infeksie van vars wonde deur luggedraagde M laxa konidia, en deur konidia en kiembuise wat reeds op die vrugoppervlak gevestig is, is gevolglik bestudeer. Pruimvrugte (kultivar Laetitia) van die pitverharding-, twee weke voor oes-, oesstadium, asook koud- \ \ opgebergde vrugte is in 'n inokulasie-toring geïnokuleer met droë M laxa konidia. .Infeksie , , van nie-gewonde vrugte en van vars wonde deur luggedraagde konidia op droë, vogtige en nat pruim vrugoppervlaktes, asook deur konidia en kiembuise wat reeds op die vrugoppervlak onder hierdie toestande gevestig is, is bepaal. Nie-gewonde groen tot ryp vrugte het meestal simptoomloos gebly, terwyl koud-opgebergde ryp vrugte wel verrot het. Wonde .het die hoeveelheid infeksie deur luggedraagde spore drasties vermeerder. Konidia wat geïnokuleer is op vrugte met vars wonde, was meer in staat om hierdie wonde te infekteer as konidia en kiembuise wat 4 dae voor wonding gevestig is. Hierdie afname in infektiwiteit was meer sigbaar op die vogtige, maar veral die nat vrugte. Hierdie studie het duidelik gewys dat inokulumvlakke op rypwordende vrugte verlaag moet word deur sanitasie-praktyke en fungisiedtoedienings. Dit is verder belangrik om vrugte, veral rypwordende vrugte, teen wonding te beskerm.

Thesis (MScAgric)--University of Stellenbosch, 2005.
ENGLISH ABSTRACT: Shrivelling and friction discolouration (FD), as postharvest disorders, negatively influence the marketability and potential shelf life of pears. By investigating the contributing factors in each of the disorders, the potential involvement of handling and storage variables were determined. This allowed for a better understanding of the responsible factors that create susceptible environments for these disorders to occur. From the moment that pears are harvested they lose weight by means of transpiration and, to a lesser extent, respiration. When excessive losses are experienced, the fruit will appear shrivelled and the marketability and shelf life are negatively influenced. By minimizing the rates of weight loss, the occurrence of shrivelling among pears during the postharvest handling can be lessened. The periods that proved to be most conducive to shrivelling (during a simulated postharvest handling duration) were where temperatures above 0 °C were experienced. These short periods proved more perilous for shrivelling than lengthy storage durations at low temperatures. This influenced the transpiration rate in such a way that the driving force accelerated the rate of weight loss in all the cultivars that were studied. Removing field heat from fresh produce and maintaining the cold chain reduces the driving force behind the transpiration of the pears. In all the cultivars studied, ‘Packham’s Triumph’, ‘Beurrè Bosc’ and ‘Forelle’, smaller and less mature fruit were more inclined to appear shrivelled. The surface area to volume ratio is fundamental in determining the rate of weight loss. This was most evident in ‘Beurrè Bosc’. Although no reproducible results could be obtained from the morphological studies, literature has attributed this phenomenon to the composition and quantity of the cuticle layer. Reduction of weight loss was obtained by sealing of the fruit stem. This obstructed water movement from the fruit through the xylem conducting tissue to the surrounding atmosphere. Not only did the stem appear greener and fresher, but less weight loss and subsequent shrivel was noticeable in the treated fruit. This effect was most evident in ‘Packham’s Triumph’ and ‘Beurrè Bosc’, but not in ‘Forelle’. ‘Forelle’ typically has a very short, thin stem in comparison to the other two cultivars.All the cultivars showed visual shrivel symptoms after 11 days at 18 °C. Rate of weight loss was the lowest in ‘Packham’s Triumph’, but due to its prominent dimensions, it appeared shrivelled before any of the other cultivars. ‘Beurrè Bosc’ lost weight at the highest rate (0.42%.day-1). As fruit injury, in the presence of oxygen, is inevitable, the oxidative enzymatic browning of pears will always be troublesome. This defensive mechanism partially prevents the infection of the fruit where epidermal cells are injured. To minimize FD, impact and frictional forces need to be lower during both harvesting and handling practices. A laboratory scale method was developed through which reproduceable treatments could be performed, thereby subjecting the fruit to industry related friction, rather than impact, injury. By assessing the discolouration in terms of both extent and intensity, the influence of variables could be determined on both ‘Packham’s Triumph’ and ‘Doyenne du Comice’ pears. As also found in practice, ‘Doyenne du Comice’ proved to be far more susceptible to FD than ‘Packham’s Triumph’, although the activity of the enzyme, polyphenol oxidase (PPO) was found to be higher in the latter. Although no significant difference was found between the FD encountered at fruit temperature of 3 °C and 15 °C, discolouration was greater at the higher temperature. This might be attributed to a greater degree of water loss, lower cell turgidity or higher enzyme activity. Thus, fruit taken from storage and sorted directly thereafter will exhibit less FD. The contribution of condensation forming on the fruit, acting as lubrication, cannot be ignored. Such fruit, with high turgor pressure, might again be more susceptible to bruising which will only be revealed well after the injury. Since enzymes, which include PPO, catalyse biochemical reactions, the availability of sufficient substrate most probably regulates the extent of this biochemical discolouration. Harvesting at optimum maturity and preventing any unnecessary friction will most definitely reduce the occurrence of FD. The ultimate challenge remains to optimize sorting and packing conditions without compromising on fruit quality.
AFRIKAANSE OPSOMMING: Verrimpeling en friksie verbruining (FV), as na-oes defekte, het ’n negatiewe invloed op die bemarkbaarheid en potensiële raklewe van pere. Hierdie studie het die bydraende faktore vir elkeen van die defekte ondersoek. Daar is gepoog om die effek van verskillende hanteringsen opbergingsmetodes op bogenoemde defekte te bepaal. ‘n Beter begrip is verkry van die oorsaaklike faktore wat bydra tot die ontstaan van die betrokke defekte. Direk na die oes van pere, begin die vrug gewig verloor as gevolg van veral transpirasie. Oormatige verliese sal lei tot ’n vrug wat verrimpeld voorkom, met ‘n negatiewe invloed op die bemarkbaarheid en raklewe daarvan. Die voorkoms van die verrimpeling van pere tydens die na-oes hantering van die vrugte, kan verminder word deur vermindering van die tempo van gewigsverlies. Die periodes waartydens die verrimpeling veral voorgekom het, (soos gevind in ’n gesimuleerde na-oes hanteringsmodel) was wanneer temperature bo 0 °C ondervind is. Sulke kort periodes was meer geneig om aanleiding te gee tot verrimpeling as die verlengde periodes van opberging by lae temperature. Sulke periodes van hoër temperature het gelei tot versnelde transpirasie en ’n versnelde tempo van gewigsverlies in al die kultivars wat ondersoek is. Die transpirasietempo van pere kan verlaag word deur die verwydering van veld-hitte en deur die streng behoud van die koue-ketting. In al die kultivars wat ondersoek is, ‘Packham’s Triumph’, ‘Beurrè Bosc’ en ‘Forelle’, is gevind dat die kleiner en minder volwasse vrugte meer geneig was tot verrimpeling. Die oppervlak area tot volume verhouding is krities in die bepaling van die tempo van gewigsverlies. Hierdie bevinding was die prominentste in ‘Beurrè Bosc’. Alhoewel geen beduidende resultate verkry kon word van die morfologiese studies nie, is daar verskeie verwysings in die literatuur wat hierdie verskynsel toeskryf aan die samestelling en hoeveelheid van die kutikula laag. Vermindering van gewigsverlies is verkry deur verseëling van die vrugtestingel. Hierdie tegniek het gelei tot ’n blokkering van die watervloei van die vrug na die omgewing deur die xileem weefsel. Verseëling van die stingel het dit groener en varser laat voorkom, en het ook ’n merkbare vermindering in gewigsverlies en die daaropvolgende verrimpeling tot gevolg gehad. Die effek van stingel-verseëling was die prominentste in ‘Packham’s Triumph’ en‘Beurrè Bosc’. Dit was minder duidelik in ‘Forelle’ wat tipies gekenmerk word deur ‘n baie korter, dun stingel in vergelyking met die ander twee kultivars. Al die kultivars het makroskopiese verrimpeling getoon na ‘n opbergingperiode van 11 dae by 18 °C. Die tempo van gewigsverlies was die laagste in ‘Packham’s Triumph’ alhoewel dit eerste verrimpeld voorgekom het. Dit kan toegeskryf word aan die spesifieke afmetings van hierdie betrokke kultivar. ‘Beurrè Bosc’ het vinnigste gewig verloor (0.42%.dag-1). Aangesien die besering van vrugte, in die aanwesigheid van suurstof, onvermydelik is, sal oksidatiewe, ensiematiese verbruining van pere altyd problematies wees. Hierdie verdedigingsmeganisme voorkom tot ’n mate die infeksie van die vrug wanneer epidermale selle beskadig word. FV kan beperk word deur die vrugte tydens oes en hanteringsprosedures so min as moontlik bloot te stel aan impak en friksie kragte. ’n Laboratorium-model is ontwikkel ter nabootsing van die omstandighede in die industrie. Die vrugte is aan friksie, eerder as impak, onderwerp, soos ondervind in die industrie. Die omvang asook die intensiteit van die verbruining is gemeet in beide ‘Packham’s Triumph’ en ‘Doyenne du Comice’ pere. Op hierdie wyse kon die invloed van die onderskeie veranderlikes in elke kultivar bepaal word. Alhoewel die ensiematiese aktiwiteit van die polifenol oksidase ensiem (PFO) die hoogste in ‘Packham’s Triumph’ was, is gevind dat ‘Doyenne du Comice’ veel meer geneig was tot FV as ‘Packham’s Triumph’. Hierdie bevinding bevestig die verskynsel soos in die praktyk gevind. Alhoewel geen betekenisvolle verskil gevind is tussen FV by vrug temperatuur van 3 °C en 15 °C nie, was daar meer verbruining by die hoër temperatuur. Hierdie verskynsel kan toegeskryf word aan ’n groter mate van waterverlies, laer sel turgiditeit en hoër ensiem aktiwiteit. Dus sal vrugte wat direk na opberging gesorteer word, minder FV toon. Die bydrae van die kondensasie wat op die vrug vorm, en as ’n smeermiddel dien, kan nie geïgnoreer word nie. Sulke vrugte met hoër turgiditeit, mag egter meer vatbaar wees vir kneusing, wat egter eers ’n geruime tyd na die kneusing tevoorverskyn mag kom. Aangesien ensieme, wat PFO insluit, as katalis dien in biochemiese reaksies, sal die beskikbaarheid van voldoende substraat, na alle waarskynlikheid die omvang van die biochemiese verkleuring reguleer.Die oes van pere tydens optimum volwassenheid, en die voorkoming van onnodige friksie sal definitief die voorkoms van FV verminder. Die uitdaging is steeds om sortering- en verpakkingstegnieke verder te verfyn sonder om ’n negatiewe invloed op vrugtekwaliteit te hê.

Field experiments were conducted in two different growing environments to evaluate the spread and movement of Dickeya dadantii. A procedure to inoculate seed potatoes with Dickeya dadantii was developed to use during this study. Spread of Dickeya dadantii from inoculated potato seed to healthy potato seed during the handling, cutting and planting procedures was not detected at either location. Spread of Dickeya dadantii from inoculated seed to surrounding progeny tubers in the field was documented in both locations. In Florida, 33% of progeny tubers tested positive for Dickeya using PCR, and in North Dakota, 13% of the progeny tubers tested positive. Stunting was observed in plants grown from Dickeya dadantii inoculated seed tubers in North Dakota, but not in Florida. These results indicate that Dickeya dadantii may spread during the seed handling and cutting processes and can spread in the field from infected seed tubers to progeny tubers.

Germinating uredospores of race C17 of Puccinia graminis f.sp. tritici form characteristic infection structures (appressorium, infection peg, vesicle, infection hypha) in response to a 1.5 h heat shock at 29° C administered 2 h after germination at 19° C. The proportion of sporelings forming infection structures was augmented by nutrients, n-nonyl alcohol and, an appropriately timed heat shock. The heat shock temperature required to induce maximum differentiation had a very precise optimum which varied slightly for each spore lot. Variations one degree above or below this optimum reduced the percent differentiation by greater than 40%. The presence of an inhibitor of protein synthesis, puromycin, in the germination medium: (1) prevented uredosporeling differentiation but had no effect on germination, (2) significantly reduced the proportion of germtubes forming appressoria, and (3) in most cases prevented the division of germtube nuclei. It was concluded that essential differentiation-specific proteins are synthesized from the onset of germination, throughout the formation of appressoria and to the completion of differentiation. These results were consistent with the observed effects of heat shock on the rate of protein hydrolysis. During germination there was a net hydrolysis of protein leading to an increase in size of the endogenous pool of free amino acids and to an increased leakage of amino acids to the germination medium. Heat shock effectively reduced the amount of endogenous free amino acids and the extent to which amino acids were lost to the medium. It was concluded that in heat shocked sporelings protein synthesis was increased relative to protein hydrolysis by comparison with the relative rates of these two processes in germinating (non-shocked) uredosporelings. Moreover, there was no net protein synthesis during the formation of infection structures induced by heat shock. The loss of amino acids to the germination medium was selective, particularly in heat shocked sporelings.
Land and Food Systems, Faculty of
Graduate

Studies presented in this dissertation highlight the importance of fungal pathogens in forestry nurseries in South Africa. Both Acacia meamsii seedlings and Eucalyptus hybrid cuttings are shown to be affected by important nursery pathogens. Chapter one presents an evaluation of the potential importance of pathogens to Eucalyptus hedge plants maintained in hydroponics. Hydroponics is a new technology being used in South African forest nurseries, which allows for the rapid establishment of Eucalyptus hedge plants. However, no information is available on pathogens affecting Eucalyptus in hydroponics. By applying information on pathogens of other hydroponic crops, several potentially important pathogens were identified and these reside in the genera Phytophthora, Pythium and Fusarium. Possible disease symptoms in Eucalyptus caused by these pathogens include wilting, stem cankers and root rots. Implementation of appropriate control measures that include cultural, biological and chemical practices could prevent and/or reduce disease impact in hydroponics. Chapter two presents the results of a survey of the roots of Eucalyptus hedge plants grown in an ebb and flow hydroponic system. An interesting result of the survey was the discovery of Cylindrocladium pauciramosum in the hydroponic system. This is the first report of the pathogen in a hydroponic system. Other important pathogens in the genera Phytophthora and Pythium were also isolated. Two Pythium species, namely P. dissotocum and P. helicoids, found in the roots and nutrient solution are new to Eucalyptus. Several Fusarium species were also isolated of which two, namely F. nygamai and F. lateritium, are also new to Eucalyptus. Chapter three of this dissertation presents the results of a survey of Eucalyptus cuttings conducted at four forestry nurseries in KwaZulu-Natal, South Africa. Several well-known Eucalyptus nursery pathogens were isolated. Cylindrocladium pauciramosum was identified as the dominant pathogen on Eucalyptus cuttings. This was confirmed based on morphological characteristics and DNA sequence comparisons. Pathogenicity tests conducted using a spore suspension of C. pauciramosum indicated that this pathogen is capable of infecting most commercial Eucalyptus clones used in South Africa. Chapter four considers a serious disease of Acacia mearnsii seedlings caused by an unidentified species of Cylindrocladium. Cylindrocladium pauciramosum was isolated from A. mearnsii seedlings showing girdling and stem canker symptoms. The pathogen was identified based on morphological characteristics and DNA sequence comparisons. Pathogenicity tests with Acacia seedlings confirmed the susceptibility of this tree to C. pauciramosum infection. This dissertation clearly indicates that Cylindrocladium pauciramosum is an important nursery pathogen in South African forestry nurseries. This pathogen has already been shown to be limiting during production of planting stock. I hope to have highlighted the importance of C. pauciramosum and other nursery pathogens in forestry nurseries in South Africa. This study will also hopefully provide information to forestry nursery managers and help them improve production.
Dissertation (MSc)--University of Pretoria, 2004.
Microbiology and Plant Pathology
Unrestricted

White mold caused by Sclerotinia sclerotiorum attacks a wide host range of broad-leafed plants which includes potatoes. Current control is limited to the use of chemicals, but biological control has emerged as an eco-friendly alternative. For the study, 19 bacterial strains and 18 fungal strains obtained from soil samples taken from a potato field naturally infested with the pathogen S. sclerotiorum were tested for the effects on S. sclerotiorum mycelium growth and sclerotia viability in vitro. A total of eight bacterial strains and six fungal strains proved to be effective in the inhibition of S. sclerotiorum mycelium growth on PDA plate using a dual culture technique. These antagonistic microbes were six Bacillus subtilis, Bacillus pumilis, Bacillus marisflavi, Fusarium solani, Fusarium equiseti, Fusarium chlamydosporum, Aspergillus niger, Aspergillus insuetus and Penicillium citrinum. There was a significant difference (p<0.05) in the interaction between S. sclerotiorum mycelium growth and the time after inoculation with a S. sclerotiorum mycelium plug for both bacterial and fungal soil isolates. An additional sclerotia viability test was carried out using four of the eight antagonistic bacteria, and six of the antagonistic fungi. The results showed that two Bacillus species, namely B. pumilis and B. marisflavi can effectively reduce sclerotia viability. The other two bacteria (both B. subtilis), recorded low percentage reduction in sclerotia viability. As for the six antagonistic fungi, the viability test proved to be less effective in determining sclerotia viability. However, the result of this study supports the use of bio-control agents, not only because they are environmentally friendly, but because they are also an effective way of controlling the plant pathogen, S. sclerotiorum.

False codling moth is a known pest of economic importance to many cultivated crops in South Africa and Africa south of the Sahara, and is particularly severe on citrus. If the fruit is infested just before harvest the chances of detecting signs of infestation are very low. As a result, the risk of packaging infested fruit and exporting them as healthy fruit is high. It is therefore a priority to develop a post-harvest technique for detection of False codling moth in citrus fruit at different levels of infestation in order to reduce phytosanitary risk. Compounds released and detected were indicative of infestation and were not insect produced but naturally produced fruit volatiles emitted at higher levels as a result of the insect within the fruit. Five major volatile compounds of interest were released by the infested oranges. These major volatile compounds include D-limonene, 3,7-dimethyl-1,3,6-octatriene, (E)-4,8-dimethyl-1,3,7-nonatriene, caryophyllene and naphthalene. Limonene was one of the most abundant volatile compounds released by the infested citrus fruit. Naphthalene, which is possibly produced due to larval feeding and development within the fruit maintained higher concentrations than controls throughout the infestation within the fruit. Naphthalene would be a good indicator of False codling moth infestation, however, not primarily for early infestation detection. A significantly higher concentration of D-limonene, 3,7-dimethyl-1,3,6-octatriene, (E)-4,8-dimethyl-1,3,7-nonatriene and naphthalene was detected using the SEP over the SPME technique. The application of an SPME procedure and the utilization of this method for detection of volatiles present in the headspace of intact infested fruit are evaluated and the possible volatile compounds diagnostic of Thaumatotibia leucotreta infestation of orange fruit and differences in volatile compound response in different orange varieties is discussed.

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).

12 pp.
plant disease bulletins; July 2006 original publishing date, rev. 10/08
Cacti, agaves and yuccas are classified as succulents, plants that have highly specialized anatomical features such as thick waxy cuticles, fleshy or minimal leaves, modified leaves (spines), and roots with extra storage capabilities for food and water. These modifications allow them to survive and thrive in harsh desert environments. They survive long periods of drought in areas of sparse rainfall and intense heat. During stressful periods, many succulents cease to grow, drop unnecessary leaves, dehydrate and become dormant until conditions for growth return. Despite their adaptations, succulents suffer from diseases, insect pests and cultural problems. Some of the more common problems that occur in cacti, agave and yuccas in Arizona are discussed in this bulletin.

Thesis (MSc)--Stellenbosch University, 2015.
ENGLISH ABSTRACT: The codling moth, Cydia pomonella (Lepidoptera: Tortricidae), is the key pest of apples and pears worldwide. The withdrawal of certain fundamental chemicals from codling moth management spray programmes, due to concerns about human, environmental and ecosystem health, has resulted in the search for softer, more environmentally friendly, and safer control measures. Entomopathogenic nematodes (EPNs), naturally occur in the soil, and actively search for hosts. The interest in using EPNs from the families Heterorhabditidae and Steinernematidae as a control measure was sparked in 1953, when an EPN was discovered in an insect. The aim is to incorporate EPNs in an integrated pest management (IPM) programme, to ensure minimal residue and eventually residue-free pome fruit production in South Africa. In order to ensure EPN success, both the environmental and technical factors influencing their efficacy, were investigated in this study. The biocontrol potential of three imported EPN isolates, being Steinernema feltiae and two isolates of Heterorhabditis bacteriophora (Hb1, Hb2), as well as a local isolate, Steinernema yirgalemense, were evaluated for the control of the codling moth under local conditions. All concentrations of S. yirgalemense, applied by immersion in a suspension of nematodes, gave > 98% control. The two formulated isolates of H. bacteriophora, Hb1-f and Hb2-f, gave < 30% control. When using the same nematode isolates, produced in vivo, S. yirgalemense still resulted in a higher codling moth control of > 90%, compared to 54% and 31% control of the H. bacteriophora Hb1 and Hb2 isolates, respectively. In follow up field trials, S. feltiae resulted in ≥ 80% control, and was thus more effective than both S. yirgalemense and the H. bacteriophora (Hb1) isolates, with 66% and 24%, and 24% and 9% control, for two separate trials, respectively. To validate the data obtained from the field trials, subsequent laboratory bioassays were conducted evaluating temperature regimes, following the same cycle as under natural conditions, with a constant humidity of 100%. Steinernema feltiae proved to be most effective, causing > 90% mortality, followed by S. yirgalemense, with 78% mortality. The two H. bacteriophora isolates (Hb1, Hb2) under the above-mentioned laboratory conditions, resulted in 73% and 59% control, respectively. Humidity thus seems to be the most important factor affecting EPN efficacy during above-ground applications. Steinernema feltiae proved to be a better candidate than S. yirgalemense for the control of the codling moth. The efficacy of different EPN isolates in controlling diapausing codling moth larvae at different temperatures was also evaluated, under local conditions, using spray application. Steinernema feltiae and two isolates of H. bacteriophora Hb1 and Hb2, including two local isolates, S. yirgalemense and Steinernema jeffreyense, were evaluated. The use of S. jeffreyense resulted in the most effective control, with 67% mortality, followed by H. bacteriophora (Hb1) with 42%, and then by S. yirgalemense with 41%. Laboratory bioassays simulating field conditions revealed that S. feltiae was most virulent to codling moth larvae, with 67% mortality by infection, followed by S. yirgalemense with 58%, the H. bacteriophora strain Hb1 with 48%, and the Hb2 strain with 24%. A comparison of the infection and penetration rate of two isolates of H. bacteriophora (Hb1, Hb2), S. feltiae and S. yirgalemense, which was carried out in multiwell plates at 14°C and 25°C, respectively, confirms the dramatic effect of temperature on EPN efficacy. At 14°C, all treatments with EPN species resulted in slower codling moth mortality than they did at 25°C, as after 48 h, < 15% mortality was recorded for all species, whereas at the warmer temperature, > 98% mortality was recorded for all species. After the exposure of washed, cool-treated larvae to 25°C for 24 h, the application of both S. feltiae and S. yirgalemense resulted in 100% mortality, whereas the application of the two H. bacteriophora isolates, Hb1 and Hb2, resulted in 68% and 54% control, respectively, over the same time period. At 14°C, S. feltiae had the highest average penetration rate of 20 IJs/insect, followed by S. yirgalemense with 14 IJs/insect, whereas S. yirgalemense had the highest penetration rate at 25°C, with 39 IJs/insect, followed by S. feltiae, with 9 IJs/insect. The two H. bacteriophora isolates had higher average penetration rates at the higher temperature. This study has highlighted the biocontrol potential of S. jeffreyense, as well as showing that S. feltiae is a cold-active nematode, whereas the other three EPN isolates prefer warmer temperatures. Stacked wooden fruit bins are regarded as preferred overwintering sites for codling moth diapausing larvae. Control strategies against the codling moth in South Africa have been hampered by the reinfestation of orchards by nearby stacked infested fruit bins or by the movement of bins between orchards. Worldwide, wooden fruit bins are systematically being replaced with plastic bins, which, in South Africa, will only be phased out over a few years. The objective of this study was to evaluate the potential of H. bacteriophora, S. feltiae, and S. yirgalemense, to disinfest miniature wooden fruit bins under controlled conditions in the laboratory. After dipping minibins in a suspension of 25 IJs/ml of all three EPN species, under optimum conditions of temperature and humidity, the highest percentage of control was obtained using S. feltiae (75%) followed by S. yirgalemense (57%), and then by H. bacteriophora (Hb1) (27%). The addition of adjuvants significantly increased (p < 0.001) S. feltiae infectivity to > 95%, whereas it did not result in a significant increase in H. bacteriophora or S. yirgalemense infectivity. The results indicated that H. bacteriophora would not be a suitable candidate to use for the control of the codling moth larvae in wooden fruit bins. The current preferred candidate for control would be S. feltiae, whose efficacy could be increased by means of the addition of an adjuvant. During winter, when the whole codling moth population are larvae and in diapause, no control measures are applied in orchards. This study has shown that EPNs can be sprayed in orchards to lower the codling moth cohort emerging after winter, as well as be included in an IPM programme. EPNs can act as a second line of defence, through supplementary control, and ensure effective control of the codling moth larvae which survived chemical spray applications, to safeguard against resistant codling moth populations in the next season.
AFRIKAANSE OPSOMMING: Kodlingmot, Cydia pomonella (Lepidoptera: Tortricidae), is ‘n belangrike plaag van appels en pere wêreldwyd. Die onttrekking van sekere fundamentele chemikalieë vanuit die kodlingmot beheerprogram weens die kommer oor menslike, omgewings en ekosisteemgesondheid, het gelei tot die soektog na sagter, meer omgewingsvriendelike en veiliger beheermaatreëls. Entomopatogeniese nematodes (EPNs) kom natuurlik in die grond voor en soek aktief na gashere. Die belangstelling in die gebruik van EPNs van die families Heterorhabditidae en Steinernematidae as 'n beheermaatreël is te danke aan die ontdekking van 'n EPN in ‘n insek in 1953. Die doel is om EPNs in 'n geïntegreerde plaagbeheerprogram (GPB) te inkorporeer om sodoende minimale residue te verseker en uiteindelik residu vrye produksie van kernvrugte in Suid-Afrika. Ten einde die sukses van EPNs te verseker, is beide die omgewings- en tegniese faktore wat hul doeltreffendheid beïnvloed in die studie ondersoek. Die biologiese beheer potensiaal van drie ingevoerde EPN isolate, Steinernema feltiae en twee Heterorhabditis bacteriophora (Hb1, Hb2) isolate, sowel as 'n plaaslike isolaat, Steinernema yirgalemense, is vir die beheer van kodlingmot onder plaaslike toestande geëvalueer. Alle konsentrasies van S. yirgalemense, wat deur indompeling in ‘n suspensie van nematodes toegedien is, het > 98% beheer tot gevolg gehad. Die twee geformuleerde isolate van H. bacteriophora, Hb1-f en Hb2-f, het < 30% beheer gegee. Met die gebruik van dieselfde nematode isolate, wat in vivo geproduseer is, het S. yirgalemense nog steeds 'n hoër kodlingmot beheer van > 90% opgelewer, in vergelyking met die 54% en 31% beheer van die H. bacteriophora Hb1 en Hb2 isolate, onderskeidelik. Steinernema feltiae het in opvolg veldproewe ≥ 80% beheer tot gevolg gehad en was dus meer effektief as beide S. yirgalemense en die H. bacteriophora (Hb1) isolate, met 66% en 24% en 24% en 9% beheer onderskeidelik in twee afsonderlike veldproewe. Om die resultate van die veldproewe te bevestig, is daaropvolgende laboratorium biotoetse uitgevoer en temperatuur regimes is geëvalueer deur die selfde siklus as onder natuurlike toestande te volg, met 'n konstante humiditeit van 100%. Die studie het bewys dat S. feltiae die mees doeltreffende isolate was met > 90% mortaliteit, S. yirgalemense het gevolg met 78% mortaliteit. Die twee H. bacteriophora isolate (Hb1, Hb2) het onderskeidelik onder bogenoemde laboratorium toestande 73% en 59% beheer tot gevolg gehad. Humiditeit blyk dus die belangrikste faktor te wees wat EPN se doeltreffendheid tydens bogrondse toediening affekteer. Die studie het bewys dat S. feltiae 'n beter kandidaat as S. yirgalemense vir die beheer van kodlingmot is. Die doeltreffendheid van verskillende EPN isolate vir die beheer van diapause kodlingmot larwes sowel as EPN se aktiwiteit by verskillende temperature is ook onder plaaslike toestande, deur bogrondse bespuitings, geëvalueer. Steinernema feltiae en twee isolate van H. bacteriophora (Hb1, Hb2), S. yirgalemense en ‘n ander plaaslike isolaat, Steinernema jeffreyense, is geëvalueer. Die gebruik van S. jeffreyense, het tot die mees effektiewe beheer gelei, met 67% mortaliteit, gevolg deur H. bacteriophora (Hb1) met 42%, en dan S. yirgalemense met 41%. Laboratorium biotoetse wat veldtoestande simuleer, het bewys dat S. feltiae die mees doeltreffend teen kodlingmot larwes is, met 67% mortaliteit tydens infeksie, gevolg deur S. yirgalemense met 58%, die H. bacteriophora Hb1 isolaat met 48%, en die Hb2 isolaat met 24%. 'n Vergelyking van die infeksie- en penetrasie tempo van twee isolate van H. bacteriophora (Hb1, Hb2), S. feltiae en S. yirgalemense wat in 12-put plate teen 14°C en 25°C uitgevoer is, het die dramatiese effek van temperatuur op EPN doeltreffendheid bevestig. By 14°C het alle EPN spesies behandelings stadiger kodlingmot mortaliteit as by 25°C na 48h tot gevolg gehad. ‘n Mortaliteit van < 15% is vir alle spesies aangeteken terwyl by die warmer temperature is > 98% mortaliteit vir alle spesies aangeteken. Na die blootstelling van afgespoelde, koel behandelde larwes aan 25°C vir 24 uur, het die toediening van beide S. feltiae en S. yirgalemense, 100% mortaliteit van larwes tot gevolg gehad terwyl die toediening van die twee H. bacteriophora isolate, Hb1 en Hb2, onderskeidelik 68% en 54% beheer tot gevolg gehad, oor dieselfde tydperk. By 14°C, het S. feltiae die hoogste gemiddelde penetrasie tempo van 20 ILs/ larwe, gevolg deur S. yirgalemense met 14 ILs/ larwe tot gevolg gehad, terwyl S. yirgalemense die hoogste penetrasie tempo getoon het by 25°C met 39 ILs/ insek, gevolg deur S. feltiae met 9 ILs/ insek. Die twee H. bacteriophora isolate (Hb1 en Hb2) het ook hoër gemiddelde penetrasie tempo by die hoër temperatuur getoon. Hierdie studie het die biobeheer potensiaal van S. jeffreyense beklemtoon, asook weereens bevestig dat S. feltiae ‘n koue-aktiewe nematode is, terwyl die ander drie EPN isolate warmer temperature verkies. Hout vrugtekratstapels, word beskou as ‘n ideale oorwintering skuiling vir kodlingmot diapause larwes. In Suid-Afrika word beheerstrategieë teen kodlingmot in die wiele gery deur die herbesmetting van boorde deur nabygeleë besmette hout vrugtekratte of deur die beweging van kratte tussen boorde. Hout vrugtekratte word wêreldwyd stelselmatig vervang met plastiek kratte. Dit sal egter eers oor ‘n aantal jare in Suid-Afrika uitgefaseer word. Die doel van hierdie studie was om die potensiaal van H. bacteriophora, S. feltiae, en S. yirgalemense te evalueer deur miniatuur hout vrugtekratte onder gekontroleerde toestande in die laboratorium te disinfekteer. Na die onderdompeling van die mini vrugtekratte in 'n nematode suspensie van 25 ILs/ml van al drie EPN spesies, onder optimale toestande van temperatuur en humiditeit, is die hoogste persentasie van beheer met die gebruik van S. feltiae (74,85% ± 3.64%) verkry. Die byvoeging van toevoegings middels het S. feltiae se vermoë om te infekteer betekenisvol (p <0,001) tot > 95% verhoog, maar dit het nie tot 'n betekenisvolle toename in die infektiwiteit van H. bacteriophora of S. yirgalemense gelei nie. Die resultate dui daarop dat H. bacteriophora nie 'n geskikte kandidaat is om te gebruik vir die beheer van kodlingmot larwes in besmette hout kratte nie. Die voorkeurkandidaat tans vir beheer is S. feltiae, waarvan die doeltreffendheid verhoog kan word deur middel van die byvoeging van 'n bymiddel. Gedurende die winter wanneer die hele kodlingmot populasie as larwes in diapause is, word geen beheer in boorde toegepas nie. Hierdie studie het getoon dat EPNs in boorde gespuit kan word om sodoende die opkomende kodlingmot populasie na die winter te verlaag en kan ook ingesluit word in 'n GPB program. Die EPNs kan as 'n tweede verdedigingslinie optree en doeltreffende beheer van kodlingmot larwes verseker wat chemiese bespuitings oorleef het, en sodoende beskerming teen weerstandige kodlingmot populasies in die volgende seisoen bied.

In this study, surveys were conducted in pea and potato fields in North Dakota and Central Minnesota to investigate the incidence and abundance of plant-parasitic nematodes in these fields. Moreover, the effect of the pin nematode, Paratylenchus nanus, on plant growth and yield of six field pea cultivars was determined under greenhouse conditions. Similarly, the influence of lesion nematode, Pratylenchus penetrans, and wilt fungi, Fusarium oxysporum alone and together on growth and yield of potato cultivar ‘Red Norland’, was evaluated in microplots under field conditions. The results indicate Paratylenchus spp. and Pratylenchus spp. are the most frequent nematodes, respectively, in pea and potato fields. Pin nematodes reproduced on field pea cultivars and caused up to 37% reduction in plant height and 40% reduction in yield. Additionally, both P. penetrans and F. oxysporum alone, and together had significant negative effect on growth and yield of potato.

Cercospora leaf spot (CLS), caused by the fungal pathogen Cercospora beticola, is the most important foliar disease of sugar beet worldwide. Control strategies for CLS rely heavily on fungicides including quinone outside inhibitor (QOI) fungicides. We collected 866 C. beticola isolates from sugar beet growing regions in France and Italy and assessed their sensitivity to the QOI fungicide pyraclostrobin. To gain an understanding of the molecular basis of QOI resistance, we cloned the full-length coding region of Cbcytb. All tested QOI-resistant isolates harbored a point mutation in Cbcytb at nucleotide position 428 that conferred an exchange from glycine to alanine at amino acid position 143 (G143A). A PCR assay was developed to discriminate QOI-sensitive and QOI-resistant isolates based on the G143A mutation. Our results indicate that QOI resistance has developed in some European C. beticola populations in Italy and monitoring the G143A mutation is an essential fungicide resistance management strategy.

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.

Dissertation (PhD (Agric))--University of Stellenbosch, 2000.
ENGLISH ABSTRACT: A study was made of the basic biology of subterranean Eriosoma lanigerum (Hausmann) populations in apple orchards in the Western Cape Province of South Africa. The absence of cornicles and the presence of a vulva could be used to identify 1st instar nymphs and adults respectively. Body length, body width and length of the hind femur are useful for separating 2nd , 3rd and 4th instars. However, separation of 2nd from 3rd instar nymphs was very unreliable. Maximum population growth rate was at 23°C while at 30 °C population growth was zero. The estimated minimum and maximum threshold temperatures for development were 4.32 and more than 30°C respectively. Numbers of underground E. lanigerum in soil samples taken using mechanical and hand augers were similar. However, numbers of aphids in samples were influenced by the distance from the trunk at which the samples were taken and the presence and the type of root material in the samples. More aphids were recorded close to the trunk, and at a given distance from the trunk more aphids were recorded if there was root material in the sample, especially if the roots were galled. early autumn (February, March) and declined during winter, especially if the winter rainfall was high. These cycles coincided with the nitrogen cycles in the roots. Embryos were also present in all instars throughout the year. There were more embryos in the 4th instar and adult aphids than in the other instars. The highest number of embryos in the 4th instar and adult aphids occurred during spring, which coincided with peak nitrogen levels in the roots of apple trees. Nitrogen levels in root material adjacent to galls and in ungalled roots were higher than in root galls. A number of entomopathogenic fungi, including species of Conidiobolus, Hirsufella and Beauveria were found. Their present contribution to biological control is not known. A straw mulch suppressed subterranean E. lanigerum population levels at least as well as the soil insecticide, imidacloprid, currently in use.
AFRIKAANSE OPSOMMING: 'n Studie van die basiese biologie van ondergrondse bevolkings van , Eriosoma lanigerum (Hausmann) is in appelboorde in die Weskaap Provinsie van Suid-Afrika uitgevoer. Die afwesigheid van kornikels en die aanwesigheid van 'n vulva kon gebruik word om die 1ste instar nimfe en volwassennes onderskeidelik te identifiseer. Liggaamslengte, liggaamsbreedte en die lengte van die agterste femur kon gebruik word om die 2de , 3de en 4de instars van mekaar te onderskei. Onderskeiding tussen 2de en 3de instar nimfe was egter baie onbetroubaar. Maksimum bevolkingsgroei het by 23°C plaasgevind, terwyl dit nul was by 30 °C. Die beraamde minimum en maksimum temperature vir ontwikkeling was by 4.32 en meer as 30°C onderskeidelik. Getalle van ondergrondse E. lanigerum in grondmonsters wat geneem is met gebruik van meganiese en hand bore was eenders. Getalle plantluise in monsters is egter beïnvloed deur die afstand vanaf die stam waarby die monsters geneem is en die teenwoordigheid van wortelmateriaal in die monsters. Meer plantluise is aangeteken as daar wortelmateriaal in die monsters was, en veral as daar galle op die wortels was. Die appelbloedluis was dwarsdeur die jaar ondergronds aktief. Bevolkingsvlakke het gedurende die vroeë somer (November, Desember) en vroeë herfs (Februarie, Maart) toegeneem, en gedurende die winter afgeneem, veral as die winterreënval hoogwas. Embrio's was ook teenwoordig dwarsdeur die jaar. Daar was meer embrio's in die 4de instar en volwasse plantluise as in die ander instars. Die hoogste aantal embrio's in die 4de instar en volwasse plantluise het in die lente voorgekom, wat saamgeval het met piek stikstofvlakke in die wortels van appelbome. Stikstofvlakke in wortelmateriaal aangrensend aan wortels en in wortels sonder galle was hoër as in wortelgalle. Talle entomopatogeniese swamme, insluitend spesies van Conidiobolus, Hirsufella en Beauvaria IS gevind. Hulle huidige bydrae tot biologiese beheer is nie bekend nie. 'n Strooideklaag het ondergrondse bevolkingsvlakke van E. lanigerum tot ten minste dieselfde mate as die grondinsektedoder, imidacloprid, wat tans in gebruik is, onderdruk.

Bibliography: leaves 81-100. Entry of inoculum into a crop and disease development in the crop cannot be prevented because spores are airborne and there is a lack of highly resistant varieties. This makes complete control of chocolate spot unlikely. It should however, be possible to improve current levels of disease control through the integration of the factors identified in the study.

Data for this study were collected by the US Forest Service and the Pennsylvania Bureau of Forestry on nearly 600 plots in central Pennsylvania. Tree and stand characteristics recorded between 1978 and 1985 include estimates of percent defoliation on individual trees. Logistic regression using maximum likelihood estimation was employed to model individual-tree mortality of 15 species in central Pennsylvania that had been defoliated by the gypsy moth. Defoliation was estimated to the nearest ten percent for individual trees. Other variables used for prediction included stand basal area and an individual-tree relative basal area index. Success ranged from no fit for three of the species to an R value (a derivation of Akaike's information criterion) of .613 for white oak. The inclusion of defoliation in the models had a varied effect. For four of the species percent defoliation was not significant. For hickory and white oak respectively, percent defoliation raised the R value by .305 and .290 percentage points. As many as five models for each species were developed: one or two models with no defoliation measure in the model and one each for one, two or three consecutive years of defoliation measures. A beta and gamma function were used to model individual· tree basal area growth for the same 15 species. The models were fit using nonlinear least squares. Variables used include the relative basal area index, stand basal area, site index and a defoliation index that incorporated three years of individual-tree, percent defoliation. The beta and gamma functions fit equally well with values of (1 - relative mean square error) ranging from .1967 to .6290. Results for both models are presented for each species. The defoliation index was a significant variable for five of the fifteen species: white, chestnut, red, and black oak and sassafras.
Master of Science

The effects of gypsy moth defoliation on acorn production (the seed source for oak regeneration), and the forest floor regeneration environment were investigated. Parameters measured included: acorn quantity, weight, development, and viability; litterfall; and forest floor depth and biomass. Nineteen plots in north-central Virginia and western Maryland on which trees suffered heavy, moderate, or no defoliation were monitored over a two-year period.

Master of Science

Deoxynivalenol (DON), a mycotoxin produced in cereal grains infected by Fusarium Head Blight produced by Fusarium graminearium and Deoxynivalenol-3-?-D-glucopyranoside (DON-3G), were studied during processing using LC-MS-MS and GC. DON reduced significantly (P<0.05) 61.8% during milling into flour. Therefore, DON was concentrated mostly in the bran and germ. DON increased 40.8% during the fermentation stage of baking. DON increased in dough more than flour and mixed dough. Milling reduced by 23.7% but fermentation did not. But bread was significantly lower in DON-3G at 0.15 ppm than flour and dough at 0.31 ppm. The baking increased DON and decreased DON-3G showing a difference in stability of the mycotoxins during processing. Enzyme hydrolysis on DON using ?-amylase, cellulase, protease, and xylanase, showed a significant increase with cellulase (20.8%), protease (11.4%), and xylanase (35.6%) compared to wheat composite. DON may be bound to the cell wall or protein component of the kernel.