Cowpeas (Vigna unguiculata (L.) Walp) is an important African indigenous legume crop for the livelihoods of many relatively poor people residing n less developed countries of the tropics. Rural families derive a nutritious food, animal feed and in income from the production of this crop. Storage of seed is certainly the most important post-harvest operation but the losses incurred are great. These losses, due to an inability to effectively control physical and biological factors, result in problems with storage insects, moisture and associated fungi. Seeds are particularly susceptible to fungal contamination when stored at high ambient temperatures and relative humidities. To determine the storage fungi associated with cowpea seeds, surface-sterilised cowpea seeds (200 seeds from each of nine cultivars) were plated out n malt extract agar. After 5-7 days incubation at 25°C, the most dominant and common fungi recorded were Alternaria spp. followed by Penicillium spp., Aspergillus flavus and A. niger. The influence of a three-year cold storage period at ± 5°C on the fungi associated with the seeds was also investigated. Alternaria, Aspergillus and Penicillium spp. appeared to dominate. Some fungal species recorded prior to cold storage were not recorded thereafter. Certain storage fungi are known to produce mycotoxins, which are secondary fungal metabolites that are toxic to both farm animals and humans, under poor storage conditions. The presence of the fusarial mycotoxins, fumonisin BI, B2 and B3 in four cowpea cultivars (Bechwana Whit, Glenda, Iron Grey, Rhino) was investigated. The samples were extracted with methanol/water (70:30 v/v) and cleaned-up on strong anion exchange solid phase extraction cartridges. High performance liquid chromatography with pre¬column derivatisation using o-phthaldialdehyde (OPA) was used for the detection and quantification of fumonisin Bl, B2 and B3. All sampIes were contaminated with FBI, with levels ranging from 81-1002 ng g-I. Fumonisin B2 and B3 were not detected in any samples. This is believed to be the first report of fumonisin BI in cowpea seeds. Since the known fumonisin-producing Fusarium species were not found in the six different Fusarium species isolated from these four cultivars, further investigations are required to determine which fungal species are species are responsible species are responsible for the FBI production. An alternative approach to the prevention and control of fungal contamination and mycotoxin production of seeds by treating cowpea seed with essential plant oils was tested. The inhibitory activity of five essential oils (thyme, clove, peppermint, soybean and peanut) was investigated, in vitro and in vivo, on five fungal species (A. flavus, A. niger, Penicillium chrysogenum, Fusarium oxysporum and F. equiseti) commonly associated with cowpea seeds and on two cowpea cultivars. Thyme and clove oil significantly inhibited the growth of all five fungal species in vitro at 500 and 1000 ppm, while peppermint oil was successful at 2000 ppm. Peanut and soybean oil did not show any significant inhibition of fungal growth. The in vivo effect of thyme, clove and peppermint oils on naturally infected seed revealed that only thyme at 1000 ppm reduced fungal growth of storage fungi in the PAN 325 cultivar. In the PAN 311 cultivar, thyme and clove oils at 1000 ppm and peppermint oil at 2000 ppm significantIy reduced growth of storage fungi. In artificially infected seed, all three oils significantly inhibited the growth of P. chrysogenum. Thyme reduced the growth of F. oxysporum and F. equisetii, whilst peppermint oil inhibited only F. oxyspomm. These oils did not seem to adversely affect the germination nor emergence of cowpea seed. The storage fungi significantly reduced percentag germination and emergence of the white (IT 93K452-1) seed but had little or no effect on the brown (CH 14) seed. Furthermore, all three oils significantly inhibited the storage fungi on the white seed, possibly increasing the percentage germination and emergence. / Dissertation (MSc (Botany))--University of Pretoria, 2006. / Plant Science / unrestricted

Yeast biocontrol agents that were applied to 'Bose' and 'Anjou' pears in the field up to three weeks prior to harvest were found to survive on the fruit at high population levels on both pear varieties. Cryptococcus infirmo-miniatus, Cryptococcus laurentii, and Rhodotorula glutinis maintained populations averaging 5 x 10��� cfu/fruit for three weeks. Candida oleophila had high initial populations, but the population size quickly declined to levels similar to the total yeast populations on untreated fruit. After a storage period of 2-4 months, fruit that were treated with C. infirmo-miniatus three weeks before harvest showed significantly lower incidence of decay at wounds than did untreated fruit. Combinations of biocontrol agents with reduced rates of the postharvest fungicides captan and thiabendazole were effective in reducing incidence and severity of blue mold decay caused by Penicillium expansum on 'Bosc' pears. Calcium chloride was also effective in combination with some biocontrol agents. Chitosan caused reductions in decay when used alone, but not when combined with most biocontrol agents. The compounds L-asparagine, L-proline, and 2-deoxy-D-glucose were not consistently effective either alone or combined with biocontrol agents. Storage of 'Bosc' and 'Anjou' pears in atmospheres with carbon dioxide concentrations of 12% or 20% for up to six weeks significantly reduced incidence and severity of gray mold decay caused by Botrytis cinerea, but decay was not reduced when the atmospheres was only 3% CO���. In contrast, the 12% or 20% CO��� atmospheres did not have significant effects on decay caused by P. expansum. Use of the biocontrol agents C. infirmo-miniatus, C. laurentii, or R. glutinis led to reductions in decay in all atmospheres, with C. infirmo-miniatus being the most consistently effective. The biocontrol products BioSave-110 (EcoScience Corp.) and Aspire (Ecogen Corp.) were less effective than the yeasts C. laurentii, R. glutinis, and C. infirmo-miniatus which were grown in the lab. / Graduation date: 1999

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.

Please read the abstract in the section 06resume.pdf of this document / Dissertation (MSc (Plant Pathology))--University of Pretoria, 2007. / Microbiology and Plant Pathology / unrestricted

Avocados are an economically important crop in South Africa and are mainly exported to Europe. As with any other tropical and subtropical crop, avocados are prone to pre- and postharvest diseases. Until recently, chemical control was the only effective measure to control fungal avocado pathogens In 1987, a Bacillus subtilis isolate was found that showed promise as a biocontrol agent in both pre- and postharvest applications to control postharvest diseases. However, over time variable results has been obtained in semi commercial trials. From the original B. subtilis isolate several subcultures have been made and used over a 15 year period in various experimental trials. The dual culture technique was used to compare the biocontrol activity of the subcultures against postharvest pathogens (Colletotrichum gloeosporioides, Phomopsis perseae, Dothiorella aromatica and Lasiodiplodia theobromae). The subcultures differed significantly in their effectiveness and genetic stability. No difference between the subcultures could be found when DNA fingerprinting using RISA PCR was used. The most effective subculture, MI-14, was used in further studies. The mode of action employed by a biocontrol agent is of utmost importance and can be used to enhance its efficacy. In a previous study it was hypothesized that antibiosis as well as competition for nutrients and space is the modes of action involved in biocontrol of B. subtilis against postharvest pathogens of avocado. The direct interaction between B. subtilis and C. gloeosporioides on avocado fruit were observed using scanning electron microscopy. Cells of B. subtilis were observed to colonize the hyphae of C gloeosporioides. In some instances, hyphal walls were lysed in the presence of B. subtilis and may be due to the presence of enzymes or antibiotic substances. Conidia of C. gloeosporioides did not germinate in the presence of B. subtilis. Diffusible inhibitory metabolites active against C. gloeosporioides were produced in vitro by B. subtilis. Inhibitory volatile substances were also produced by B. subtilis and were found to be active against P. perseae, D. aromatica and L. theobromae but not C. gloeosporioides. Siderophores production as well as chitinase, amylase, lipase and proteinase activity were also observed and may play a role in antagonism. Antibiotic production by B. subtilis is a well-known phenomenon. Most antibiotics are polypeptides and lipopeptides. The involvement of phenolic metabilites in biocontrol by B subtilis is less known. A seven-day-old culture of B. subtilis in a minimal medium was analyzed for the presence of free acid phenolic compounds active against fungi. Free acid phenolic metabolites were found and separated using layer chromatography. TLC plates containing the separated spots were sprayed with Clasdosporium cladosporioides and plates were observed for inhibition zones. The phenolic substances were present at 7.06 ± 0.95 mg gallic acid ml-1. The phenolic substances fall in the hydroxycinamic acid group due to their fluorescent coloring under UV at 350 nm. The mode of action involved is also influenced by environmental factors. The effect of temperature and carbon- and nitrogen sources of the in vitro inhibitory activity of B. subtilis against C. gloeosporioides, P. perseae, D. aromatica and L. theobromae were investigated using the dual culture technique. The most effective temperature range for B. subtilis was found to be between 20 and 37°C. At temperatures lower than 15°C, B. subtilis was found to be not very effective, suggesting why postharvest applications followed directly by cold storage do not always work effectively. D-arabinose and D-(+)-mannitol evaluated as carbon source as well as L-glutamic acid, L-glutamine and L-(+)-asparagine used as nitrogen sources support in vitro antagonism against the pathogens most effectively. They also do not support the growth of C. gloeosporioides, P. perseae, D. aromatica and L. theobromae. These nutrients can potentially be the most effective ones to incorporate in commercial B. subtilis formulations. The study showed the potential role of antagonistic free acid phenolic substances, volatiles and siderophores on inhibition of fungal avocado pathogens. Further studies to confirm their in situ activity are required. In conclusion, various factors affect the efficacy of B. subtilis against postharvest pathogens of avocado. These factors should be kept in mind when applying the commercial product in order to achieve the best results. / Dissertation (MSc (Microbiology))--University of Pretoria, 2011. / Microbiology and Plant Pathology / unrestricted

No description available.

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.

The South African fresh fruit industry is a significant exporter, accounting for approximately 45% of the country’s agricultural exports. Of the total exported fruit in the 2005/06 season, 60% was subtropical fruit. However, certain physiological rind disorders such as mango lenticel damage and citrus rind pitting are frequently observed, reducing the commercial value of the fruit. This thesis deals with the epidemiology of these rind physiological disorders, in an attempt to obtain basic information that could serve as a guideline to predict and manage the fruit susceptibility to these disorders. The study further investigated the relative effects that certain pre-harvest factors have on the postharvest development of these disorders. Factors of particular interest were harvest maturity, climate and the mineral content of the fruit. Rind pitting is a physiological disorder of citrus that develops during storage. A study was conducted to investigate the relative effects that certain pre-harvest factors have on the post-harvest development of superficial rind pitting in ‘Benny Valencia’ oranges. Factors of particular interest were harvest maturity, climate and the mineral content of the fruit. In addition, trees were treated with two different formulations of nitrogen viz. limestone ammonium nitrate (LAN) and a slow release nitrogen fertilizer (Horticote®), during March 2006. The fruit were then sampled on a two-weekly basis over a period of three months. On each sampling date a set of quality related readings, such as juice sugar and titratable acid concentration were taken, after which the mineral concentration of the exocarp and mesocarp was measured. Fruit were also stored under export simulation conditions. The results indicate that fruit from trees that received additional N were more susceptible to rind pitting than those from control trees. Of the two N applications, fruit from trees that received slow release N were more susceptible to rind pitting than fruit from trees that received the LAN treatment. Another important observation made was that the nitrogen concentration of oranges from trees that received extra nitrogen fertilizer was lower than that from the controls. In addition, experimental fruit were smaller than control fruit. Fruit from trees that received the slow release nitrogen treatment were smallest. A sink/source hypothesis aimed at explaining this phenomenon has been formulated and is currently being investigated. A number of control mechanisms are also being explored. Mango lenticel damage is a serious defect that occurs on the rind of the fruit. This condition causes fruit to attain a speckled appearance and become unattractive to the buyer. Each season, the disorder reduces the packout of fresh fruit by about 16%. Several preharvest factors play a critical role in the postharvest development of lenticel damage. Preliminary studies have shown that the incidence of lenticel damage has some relationship to fruit moisture and fruit nutrient concentration, depending on which of these factors first achieve a critical threshold. A study was conducted to develop appropriate skin moisture content parameters to predict lenticel damage potential before harvest. The study further aimed to provide certain biological markers regarding appropriate nitrogen fertilization practices to reduce lenticel damage. To do this, two trials were conducted approximately a month before harvest. Firstly, a plastic ground cover to restrict water supply was laid under ‘Tommy Atkins’ trees at Numbi Estates in the Hazyview area of the Mpumalanga province, South Africa. Secondly, additional nitrogen was applied as three different formulations, viz. limestone ammonium nitrate (LAN), potassium nitrate (KNO3), and a slow release nitrogen fertilizer under trade name Horticote®, to two ‘Tommy Atkins’ and two ‘Keitt’ orchards at Bavaria Estates in the Hoedspruit area of the Limpopo province, South Africa. Fruit samples were harvested, packed and stored at different intervals after rain during January 2006. With regard to lenticel damage on ’Tommy Atkins’ fruit from Numbi, there was no significant difference between the control and plastic cover treatments. However, when compared to fruit from other localities, the Numbi fruit had the highest incidence of lenticel damage, followed by the conventional orchard at Bavaria. The Numbi fruit, which had the highest incidence of lenticel damage, also had the highest pulp and skin nitrogen concentration. Of the four treatments at Bavaria, the LAN treatment had the highest incidence of lenticel damage, but the pulp and skin nitrogen concentrations of this treatment were comparable with the other treatments. The organic fruit had significantly lower lenticel damage incidence and also had the lowest pulp and rind nitrogen concentrations. Similarly, ‘Keitt’ results showed that the intensity of lenticel damage was significantly higher in the orchard that received additional nitrogen in the form of LAN. From the results it was, however, not possible to formulate nitrogen-lenticel damage correlations. The study failed to prove the case for a direct relationship between the disorder and with nitrogen, as there were no significant or consistent correlations with nitrogen content. An interesting relationship was nevertheless observed between lenticel damage and the time of harvest before and after rainfall. The results indicated that both ‘Tommy Atkins’ and ‘Keitt’ fruit become more susceptible to lenticel damage when harvested a day after rainfall and this gradually reduces afterwards. The effect of rainfall in this regard and a sink-source hypothesis arising from these observations are also discussed. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2008.

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.

Neofabraea is a genus of an important plant pathogenic fungi having worldwide appearance. Four Neofabraea species are responsible for bull's eye rot of pome fruits. The aim of this thesis was to investigate which of these species occur in the Czech Republic. 81 isolates were collected during a two- year period and they were identified using PCR fingerprinting (primers ERIC 1R and M13-core) and DNA sequencing (ITS, mtSSU and tub2). The results showed that species N. alba, N. perennans and Cryptosporiopsis kienholzii occur in the Czech Republic. According to available information, this is the second record of C. kienholzii in Europe. One isolate (KP4) failed to be identified as any of the species. KP4 is very close to C. kienholzii, but it can be distinguished both biologically and genetically. Furthermore, the aggressiveness of individual species was compared.N. perennans and strain KP4 proved to be most aggressive, the least aggressive is C. kienholzii. Two N. alba strains (KP36 and KP37) isolated from healthy apple fruit and leaf are pathogenic for apple fruits. Keywords: aggressiveness, Cryptosporiopsis kienholzii, Dermateaceae, Helotiales, apple tree, Malus, PCR fingerprinting, postharvest diseases Powered by TCPDF (www.tcpdf.org)