Crown gall disease is an agricultural problem caused by the soil-borne bacterium, Agrobacterium tumefaciens. A. tumefaciens oncogenes cause transformed plant cells to overproduce the hormones, auxin and cytokinin. High hormone levels cause unorganized plant cell growth resulting in a gall. Control of crown gall disease is difficult because after plant cell transformation has occurred, the bacterium is no longer required for the disease to progress. Apple trees engineered to express double-stranded RNA of two A. tumefaciens oncogenes, ipt and iaaM, silenced the expression of the wild-type oncogenes and prevented crown gall disease. Only the iaaM oncogene was targeted for posttranscriptional gene silencing (PTGS) as measured by biological assays and by quantitative reverse-transcription polymerase chain reaction (q-RTPCR) on transgenic tissue. However, if the translation initiation sequence of the iaaM construction was eliminated, gall formation was not prevented, indicating that translatable RNA initiates silencing whereas untranslatable RNA does not. Other data indicate that the Arabidopsis thaliana micro-RNA pathway gene is involved in A. tumefaciens-mediated tumorigenesis. A. thaliana plants with a mutation in HEN1, a gene required for micro-RNA maturation, demonstrated a tenfold reduction in tumorigenesis upon A. tumefaciens infection compared to wild-type. The same plant line showed no difference in T-DNA transfer and nuclear uptake. / Graduation date: 2006

Thesis (MscAgric (Horticulture))--University of Stellenbosch, 2007. / Production of quality fruit is the main aim in horticultural crops. Numerous research reports stress the important role of calcium (Ca) in maintaining firmness and preventing the development of physiological disorders in fruit. This study focused on the effect of water and nutrient management strategies, rootstocks and foliar Ca applications on fruit Ca content. Final Ca content/concentration in apple fruit at harvest did not differ significantly between treatments water with micro jets (hand nutrition), water and nutrients with fertigation, or water and nutrients with pulsating drip when applied to ‘Brookfield Gala’ trees in third leaf, on two rootstocks (M793 and M7). In the second trial, three Ca levels were applied to ‘Brookfield Gala’ trees through a pulsating drip system during three phenological periods to evaluate the effect on Ca uptake of the fruit. During the second season, application of high Ca levels for the period full bloom to harvest gave a higher fruit Ca concentration than with applications of standard or low Ca.

Thesis (MScAgric (Horticulture))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: Peel colour is an important quality factor in the production of bi-coloured apple fruit. Most markets set minimum requirements for red colour coverage. Fruit that do not meet these requirements are downgraded and has a major impact on the profitability of apple production in South Africa. South African apple production areas are amongst the warmest in the world. Since anthocyanin accumulation requires induction at low temperature and synthesis require mild temperatures, experiments were conducted to investigate optimum day and night temperatures for red colour development throughout fruit development for red and bi-coloured apple cultivars grown in South Africa. We found that redder strains of bi-coloured apple cultivars did not appear to owe their enhanced pigmentation to higher temperature optima for anthocyanin synthesis. The optimum day temperatures for red colour development in the different cultivars seemed to differ between seasons, but not between production areas. In general, red colour in the cultivars evaluated developed maximally between 17 ºC and 25 ºC. The optimum day temperature for red colour development remained constant throughout fruit development for most cultivars, but increased roughly from 14 ºC to 22 ºC in ‘Cripps’ Pink’ between January and April. The extent of red colour development increased during fruit development in all the cultivars assessed. We were unable to determine optimum induction temperatures for red colour development. ‘Royal Gala’ from Ceres seemed to benefit from induction at 4 ºC while red colour in ‘Fuji’ decreased with decreasing temperature. To explain the presence of anthocyanins in immature apple fruit, we tested the hypothesis that anthocyanins protect the peel from photoinhibition and photooxidative damage during conditions of increased light stress. First we established that the rate of colour change in response to a passing cold front appears to be sufficient to provide photoprotection during a cold snap. Also in agreement with the hypothesis, ‘Cripps Pink’ peel incurred significantly more photoinhibition at low temperature (16 ºC) compared to mild (24 and 32 ºC) and high (40 ºC) temperature under high irradiance with visible light. Recovery rate was temperaturedependent, being the slowest at low temperature and increasing with temperature. The photoapparatus in ‘Cripps Pink’ peel appears to be particularly sensitive to light stress at low temperature throughout the season, with significant photoinhibition occurring even at moderate temperature (24 ºC). The sensitivity of the apple peel to photoinhibition increased throughout the season at lower irradiance levels, but remained the same at higher irradiance. In our final experiment, fruit were exposed to high irradiance at low and mild temperature before exposure to high temperature in combination with high irradiance. This was done to test the hypothesis that photoinhibition incurred during cold snaps predisposes peel to photothermal damage when temperature increases again after the cold snap. Unfortunately, due to the severity of the stress incurred in response to high temperature treatment, the results were inconclusive. / AFRIKAANSE OPSOMMING: Vrugkleur is ‘n belangrike kwaliteitsfaktor in die produksie van tweekleurappels. Die meeste markte stel minimum vereistes vir rooi kleurbedekking. Vrugte wat nie aan hierdie vereistes voldoen nie, word afgegradeer. Suid-Afrika se appel produksie areas word beskou as van die warmste ter wêreld. Antosianien akkumulasie benodig induksie by lae temperature gevolg deur sintese in lig by matige temperature. Gevolglik het swak rooi kleurontwikkeling onder plaaslike toestande ‘n groot impak op die winsgewendheid van appelproduksie in Suid-Afrika. Eksperimente is uitgevoer om die optimum dag- en nagtemperature vir rooi kleurontwikkeling tydens vrugontwikkeling vir die rooi en tweekleur appel kultivars wat in Suid-Afrika geproduseer word te bepaal. Ons het gevind dat die verhoogde pigmentasie van rooier seleksies van tweekleurappel kultivars nie aan ‘n hoër temperatuur optimum vir antosianiensintese toegeskryf kan word nie. Die optimum dag temperature vir rooi kleurontwikkeling vir die onderskeie kultivars verskil klaarblyklik tussen seisoene, maar nie tussen produksie areas nie. Oor die algemeen het kleurontwikkeling maksimaal plaasgevind tussen 17 ºC en 25 ºC. Die optimum dagtemperatuur vir rooi kleurontwikkeling het konstant gebly tydens vrugontwikkeling, buiten vir ‘Cripps’ Pink’ waar dit toegeneem het van ongeveer 14 ºC tot 22 ºC vanaf Januarie tot April. Die mate van rooi kleurontwikkeling het in al die kultivars toegeneem deur die loop van vrugontwikkeling . Ons kon nie daarin slaag om optimum induksie temperature vir rooi kleurontwikkeling vas te stel nie. Rooi kleurontwikkeling van ‘Royal Gala’ uit Ceres is moontlik bevorder deur induksie by 4 ºC, terwyl ‘Fuji’ se rooi kleur afgeneem het met ‘n verlaging in induksie temperatuur. Ten einde die teenwoordigheid van antosianien in onvolwasse appelvruggies te verduidelik, het ons die hipotese getoets dat antosianien die vrugskil beskerm teen fotoinhibisie en fotooksidatiewe beskadiging gedurende tydperke van verhoogde ligstres. Eerstens het ons bevestig dat die tempo van kleurontwikkeling in reaksie op ‘n koue front waarskynlik vinnig genoeg is om fotobeskerming te verleen. Vervolgens is gevind dat ‘Cripps’ Pink’ vrugskil aansienlik meer fotoinhibisie ervaar het by lae temperatuur (16 ºC) in vergelyking met matige (24 ºC en 32 ºC) en hoë (40 ºC) temperatuur onder hoë irradiasie met sigbare lig. Die hersteltempo was temperatuur-afhanklik; dit was die stadigste by lae temperatuur en het toegeneem met ‘n toename in temperatuur. Die foto-apparaat in ‘Cripps’ Pink’ vrugskil blyk besonder sensitief te wees vir ligstres by lae temperatuur regdeur die groeiseisoen met aansienlike fotoinhibisie by selfs matige temperatuur (24 ºC). Die sensitiwiteit van die vrugskil vir fotoinhibisie het toegeneem deur die groeiseisoen by laer ligvlakke, maar het dieselfde gebly by hoër vlakke van irradiasie. Laastens is vrugte blootgestel aan hoë irradiasie by lae en matige temperatuur voordat dit vervolgens blootgestel is aan hoë temperatuur in kombinasie met hoë irradiasie. Dit was om die hipotese te toets dat fotoinhibisie wat opgedoen word gedurende ‘n onverwagte koue periode, die skil meer vatbaar maak vir fototermiese skade sodra die temperatuur weer styg na die koue periode verby is. Ongelukkig het die hoë temperatuur stres al die behandelings tot so ‘n mate geaffekteer dat dit onmoontlik was om enige gevolgtrekkings vanuit ons resultate te maak.

Doctor of Philosophy (PhD) / The goal of this research was to investigate changes in the physiological properties including firmness, stiffness, weight, background colour, ethylene production and respiration of ‘Cripps Pink’ apple stored under different temperature and atmosphere conditions,. This research also seeks to establish mathematical models for the prediction of changes in firmness and stiffness of the apple during normal atmosphere (NA) storage. Experiments were conducted to determine the quality changes in ‘Cripps Pink’ apple under three sets of storage conditions. The first set of storage conditions consisted of NA storage at 0oC, 2.5oC, 5oC, 10oC, 20oC and 30oC. In the second set of conditions the apples were placed in NA cold storage at 0oC for 61 days, followed by NA storage at the aforementioned six temperatures. The third set of conditions consisted of controlled atmosphere (CA) (2 kPa O2 : 1 kPa CO2) at 0oC storage for 102 days followed by NA storage at the six temperatures mentioned previously. The firmness, stiffness, weight loss, skin colour, ethylene and carbon dioxide production of the apples were monitored at specific time intervals during storage. Firmness was measured using a HortPlus Quick Measure Penetrometer (HortPlus Ltd, Hawke Bat, New Zealand); stiffness was measured using a commercial acoustic firmness sensor-AFS (AWETA, Nootdorp, The Netherlands). Experimental data analysis was performed using the GraphPad Prism 4.03, 2005 software package. The Least-Squares method and iterative non-linear regression were used to model and simulate changes in firmness and stiffness in GraphPad Prism 4.03, 2005 and DataFit 8.1, 2005 softwares. The experimental results indicated that the firmness and stiffness of ‘Cripps Pink’ apple stored in NA decreased with increases in temperature and time. Under NA, the softening pattern was tri-phasic for apples stored at 0oC, 2.5oC and 5oC for firmness, and at 0oC and 2.5oC for stiffness. However, there were only two softening phases for apples stored at higher temperatures. NA at 0oC, 2.5oC and 5oC improved skin background colour and extended the storage ability of apples compared to higher temperatures. CA during the first stage of storage better maintained the firmness and stiffness of the apples. However, it reduced subsequent ethylene and carbon dioxide (CO2) production after removal from storage. Steep increases in ethylene and CO2 production coincided with rapid softening in the fruit flesh and yellowing of the skin background colour, under NA conditions. The exponential decay model was the best model for predicting changes in the firmness, stiffness and keeping quality of the apples. The exponential decay model satisfied the biochemical theory of softening in the apple, and had the highest fitness to the experimental data collected over the wide range of temperatures. The softening rate increased exponentially with storage temperature complying with the Arrhenius equation. Therefore a combination of the exponential decay model with the Arrhenius equation was found to best characterise the softening process and to predict changes in the firmness and stiffness of apples stored at different temperatures in NA conditions.

Doctor of Philosophy (PhD) / The goal of this research was to investigate changes in the physiological properties including firmness, stiffness, weight, background colour, ethylene production and respiration of ‘Cripps Pink’ apple stored under different temperature and atmosphere conditions,. This research also seeks to establish mathematical models for the prediction of changes in firmness and stiffness of the apple during normal atmosphere (NA) storage. Experiments were conducted to determine the quality changes in ‘Cripps Pink’ apple under three sets of storage conditions. The first set of storage conditions consisted of NA storage at 0oC, 2.5oC, 5oC, 10oC, 20oC and 30oC. In the second set of conditions the apples were placed in NA cold storage at 0oC for 61 days, followed by NA storage at the aforementioned six temperatures. The third set of conditions consisted of controlled atmosphere (CA) (2 kPa O2 : 1 kPa CO2) at 0oC storage for 102 days followed by NA storage at the six temperatures mentioned previously. The firmness, stiffness, weight loss, skin colour, ethylene and carbon dioxide production of the apples were monitored at specific time intervals during storage. Firmness was measured using a HortPlus Quick Measure Penetrometer (HortPlus Ltd, Hawke Bat, New Zealand); stiffness was measured using a commercial acoustic firmness sensor-AFS (AWETA, Nootdorp, The Netherlands). Experimental data analysis was performed using the GraphPad Prism 4.03, 2005 software package. The Least-Squares method and iterative non-linear regression were used to model and simulate changes in firmness and stiffness in GraphPad Prism 4.03, 2005 and DataFit 8.1, 2005 softwares. The experimental results indicated that the firmness and stiffness of ‘Cripps Pink’ apple stored in NA decreased with increases in temperature and time. Under NA, the softening pattern was tri-phasic for apples stored at 0oC, 2.5oC and 5oC for firmness, and at 0oC and 2.5oC for stiffness. However, there were only two softening phases for apples stored at higher temperatures. NA at 0oC, 2.5oC and 5oC improved skin background colour and extended the storage ability of apples compared to higher temperatures. CA during the first stage of storage better maintained the firmness and stiffness of the apples. However, it reduced subsequent ethylene and carbon dioxide (CO2) production after removal from storage. Steep increases in ethylene and CO2 production coincided with rapid softening in the fruit flesh and yellowing of the skin background colour, under NA conditions. The exponential decay model was the best model for predicting changes in the firmness, stiffness and keeping quality of the apples. The exponential decay model satisfied the biochemical theory of softening in the apple, and had the highest fitness to the experimental data collected over the wide range of temperatures. The softening rate increased exponentially with storage temperature complying with the Arrhenius equation. Therefore a combination of the exponential decay model with the Arrhenius equation was found to best characterise the softening process and to predict changes in the firmness and stiffness of apples stored at different temperatures in NA conditions.

Thesis (MScAgric)--University of Stellenbosch, 2003. / ENGLISH ABSTRACT: Various aspects of the biology and ecology of woolly apple aphid, Eriosoma lanigerum, were investigated, including initial galling damage caused by E. lanigerum to the roots of apple trees, the possible relationship between E. lanigerum and Xiphinema and Pratylenchus nematodes and the effectiveness of Biostart 2000® and Furfural® as possible control agents of E. lanigerum in the orchard. Preliminary root damage by first instar E. lanigerum feeding was characterized by the mechanical injury of endodermis and parenchyma tissues. Damage by second, third and fourth instar E. lanigerum was similar, but the symptoms were more pronounced. Damage caused by adults included a pronounced swelling at infected areas of the root. Cell walls hardened until the root was radially strengthened with sclerenchyma tissue and nonconducting xylem vessels while the cuticle expanded greatly through the growth of corklike cambium tissue. There was no direct relationship between the population dynamics of E. lanigerum and those of Xiphinema and Pratylenchus nematodes. The occurrence of E. lanigerum appeared to be seasonal while P. penetrans and Xiphinema numbers fluctuated erratically. Undamaged root nitrogen levels seemed to correspond with the normal root growth cycle. Nitrogen levels from galled roots were significantly lower than those of undamaged roots, probably due to E. lanigerum feeding. Soils rich in fine sand and clay sustained higher populations of E. lanigerum and Xiphinema than sandy soils. The number of E. lanigerum found in soil samples correlated well with the damage index allocated to the samples. The numbers of Xiphinema found in soil samples also correlated well with the damage index allocated to the samples according to suspected Xiphinema damage symptoms. Both Biostart 2000® and Furfural® were effective as control agents of woolly apple aphid. Furfural'Ï, a chemical waste product of the sugarcane industry, was however not as effective as Biostart 2000®, a product that includes an activator and three bacterial species, Bacillus laterosporus, B. chitinosporus and B. licheniformis. The bacteria in the Biostart 2000® treated pots could replicate themselves under suitable conditions while Furfural® dilutes with each watering. Biostart 2000® is also easier to prepare than Furfural® since the components of Biostart 2000® readily mix to form a paste easily thinned by water, whereas Furfural® is an oily substance that does not easily disperse in water. Root damage was initiated soon after E. lanigerum started feeding, however there was no apparent relationship between E. lanigerum and the nematode species. The most promising, environmentally friendly control measure was Biostart 2000®. / AFRIKAANSE OPSOMMING: Verskeie aspekte van biologie en die ekologie van die appel bloedluis, Eriosoma lanigerum, was ondersoek insluitende aanvanklike galvorming veroorsaak deur E. lanigerum op wortels van appelbome, die moontlike verwantskap tussen E. lanigerum en Xiphinema en Pratylenchus nematodes en die effektiwiteit van Biostart 2000® en Furfural® as moontlike beheeragente van E. lanigerum in die boord. Aanvanklike wortelskade deur eerste ins tar E. lanigerum voeding was gekenmerk deur die meganiese beskadiging van endodermale en parenchiem weefsel. Skade veroorsaak deur tweede, derde en vierde instar E. lanigerum was soortgelyk alhoewel die simptome meer beklemtoond was. Skade deur volwassenes het 'n meer duidelike swelsel by geïnfekteerde wortelareas ingesluit. Selwande het verhard totdat die wortel radiaalsgewys versterk was met skierenchiem weefsel en nie-geleidende xileemvate terwyl die kutikula grootliks toegeneem het deur die groei van kurkagtige kambiumweefsel. Daar was geen direkte verwantskap tussen die bevolkingsdinamika van E. lanigerum en dié van Xiphinema en Pratylenchus nematodes nie. Die voorkoms van E. lanigerum was seisoenaal terwyl P. penetrans en Xiphinema se getalle onvoorspelbaar gefluktueer het. Onbeskadigde wortel stikstofvlakke het ooreengestem met die normale wortel groeisiklus. Stikstof vlakke van galwortels was noemenswaardig laer as dié van onbeskadigde wortels, heel waarskynlik as gevolg van voeding deur E. lanigerum. Grond ryk aan fyn sand en klei het groter bevolkings van E. lanigerum en Xiphinema onderhou as sanderige gronde. Die aantal E. lanigerum in grondmonsters het goed ooreengestem met die skade indeks wat aan die monsters toegeken was. Die aantal Xiphinema in grondmonsters het ook goed ooreengestem met die beskadigingsindeks wat aan die monsters toegeken is weens vermoedelike Xiphinema skade simptome. Beide Biostart 2000® en Furfural® was effektief as beheeragente van die appelbloedluis. Furfural'", 'n afvalproduk van die suikerriet industrie, was egter minder effektief as Biostart 2000®, 'n produk bestaande uit 'n aktiveerder en drie bakterie spesies, Bacillus laterosporus, B. chitinosporus en B. licheniformis. Die bakterië in die Biostart 2000® behandelde potte kon vermeerder onder gunstige toestande terwyl Furfural® na elke besproeiing verdun het. Biostart 2000® is ook makliker om aan te maak as Furfural® aangesien die bestanddele van Biostart 2000® geredelik meng tot 'n wateroplosbare pasta, terwyl Furfural® 'n olierige vloeistofis wat moeilik 'n waterige suspensie vorm. Wortelskade het plaasgevind kort nadat E. lanigerum begin voed het, alhoewel daar geen duidelike verwantskap tussen E. lanigerum en nematode spesies voorgekom het nie. Die mees belowende omgewingsvriendelike beheermaatreël was Biostart 2000®.

Objectivity is often emphasized as one of the fundamental principles for the auditing profession and is also a requirement for preserving satisfactory professional practice. The multiple company scandals in the first decade of the 21st century have created distrust for auditors’ capability to maintain objectivity. The purpose of this study is to contribute with a new way of thinking when it comes to the factors that affect objectivity. Earlier studies have indicated that personality factors may affect work performance in many different fields and this study is exploring the connection between separate personality factors, and the capability of less experienced auditors to maintain objectivity in the relationship with dominating and demanding clients. Data was collected through a survey that was divided into two separate parts where the first part measured the participant’s score on different personality factors, and the other part measured the participant’s score on perceived objectivity. 133 subjects participated in the study and these subjects were divided into non-authorized or non-approved auditors and recently authorized or approved auditors. The result of the study did not indicate any significant connection between perceived objectivity and the different personality factors. However, there was a statistically significant connection between authorization and perceived objectivity. Non-authorized and non-approved auditors in the study tended to score higher on perceived objectivity than did their authorized counterparts. In order to regain the trust for auditors and to strengthen the role of the profession it is important to increase objectivity. The surprising evidence supplied by this study creates a curiosity and urge to further investigate the factors affecting objectivity, especially in combination with possible psychological aspects. This study is unique because it focuses strictly on individual instead of contextual factors and it thereby contributes to the literature about the influence of individual factors as well as to the study of objectivity among auditors. This is a step towards greater understanding of the factors that actually affect objectivity and, as a consequence, also affect the quality of the audit as a control mechanism. / Objektivitet framhålls ofta som en av de grundläggande principerna för revisorsyrket samt ett krav för att upprätthålla god yrkessed. Det tidiga 2000-talets stora företagsskandaler har dock skapat en misstro till revisorer som yrkesgrupp och har dessutom föranlett ett tvivel gentemot revisorers objektivitet. Avsikten med denna studie är att bidra med nya tankebanor kring vad som skulle kunna påverka objektiviteten. Tidigare studier har indikerat att personlighetsfaktorer kan påverka arbetsprestationen inom vitt skilda branscher och denna studie undersöker sambandet mellan olika personlighetsfaktorer och mindre erfarna revisorers förmåga att bibehålla objektiviteten i relationen med dominerande och krävande klienter. Data till studien samlades in genom en tvådelad enkät där en del mätte personligheten och den andra delen mätte graden av objektivitet. I studien deltog 133 respondenter fördelat på revisorsassistenter, revisorer som varken är auktoriserade eller godkända samt revisorer som har auktoriserats det senaste året. Resultatet av studien påvisar inte några signifikanta samband mellan graden av objektivitet och de studerade personlighetsfaktorerna. Istället påvisades ett signifikant samband mellan auktorisation och graden av objektivitet. Revisorer som varken var godkända eller auktoriserade visade i studien tendenser till högre grad av objektivitet än sina auktoriserade kollegor. I fråga om att vinna tillbaka förtroendet för revisorerna och att stärka deras yrkesroll är det viktigt att öka graden av objektivitet. Informationen som den här studien ger är överraskande och skapar framför allt en nyfikenhet och forskarlust att gräva djupare i objektivitetens påverkansfaktorer, och då i synnerhet i kombination med tänkbara psykologiska aspekter. Denna studie är unik då den i allra högsta grad fokuserar på individfaktorer istället för kontextuella faktorer och därmed bidrar till litteraturen om individfaktorers påverkan såväl som till studiet av objektivitet hos revisorer.  Detta är ett led i förståelsen av vilka faktorer som faktiskt har inverkan på objektivitet och därmed också har inverkan på kvaliteten hos revisionen som kontrollfunktion.

Submitted in fulfillment of the Master’s Degree in Homoeopathy, Durban University of Technology Durban, 2014. / The purpose of this research study was to determine any therapeutic significance of Malus domestica (domestic apple) in the potentised, homoeopathic form and to contribute this information to the body of the homoeopathic materia medica. It was further hypothesised that some proving symptoms experienced by the provers during the research study would show a resemblance to unique characteristics of the plant in terms of its natural appearance and cultural references. This involved a detailed doctrine of signatures analysis of Malus domestica based on an extensive literature review. Combining a proving research study with comparative analysis to the doctrine of signatures helped to clarify and verify the remedy’s potential therapeutic value. Methodology This homoeopathic proving was carried out in the form of a double blind placebo controlled study of Malus domestica 30CH with a total of 30 provers. The prover sample was randomly divided into two groups: 24 provers (80%) into the verum group and the remaining six provers (20%) into the placebo group. The provers were unaware of either the proving substance or the potency used. Participants were required to record their mental, emotional and physical status’ one week prior to administration of the proving powders as a form of control for comparison of symptoms post administration of the proving remedy. Thorough physical examinations and case histories of each prover were taken prior to and after the proving period. Provers ingested one powder three times a day for three days and recorded their symptoms daily in a journal. The duration of the proving period spanned five weeks. During this period researchers were in constant contact with all participants. Once the proving period was complete, all journals were gathered and the information therein translated into materia medica and repertory format so as to develop the remedy picture of Malus domestica 30CH. A comparison between the symptomatology produced in the provers and the doctrine of signatures was then performed. Results A wide range of symptoms were documented by the provers. The results revealed an affinity to the mental plane – the most striking being the theme of being disconnected and separated from others in the environment as well as within one’s self. Marked polarities were depicted as follows: Anger/irritability vs. calmness/tranquility; Depression/sadness vs. cheerfulness; Confusion vs. concentration/clarity of mind; Connection vs. disconnection. Physically, symptoms concerning the extremities were numerous, with provers describing symptoms of their limbs being disconnected/separated as well. Head symptoms were also numerous as provers described a wide range of headaches, as well as headaches that were associated with eye symptoms. Abdominal symptoms manifesting as cramping associated with diarrhoea and, in some cases, constipation were documented. Dizziness associated with the sensation of the head floating was common as well. Numerous themes arising from dreams were recorded by provers, with a clear affinity to family members as well as dreams of social gatherings and banquets. Provers dreamt of family members placed in precarious situations wherein the lives of their loved ones were at risk, whilst the emotion of guilt was expressed in certain dreams regarding parties and banquets. The comparison between the homoeopathic drug proving of Malus domestica 30CH and the doctrine of signatures brought many similarities to light, with most of these similarities relating to the mind, extremities and head. Conclusion As hypothesised Malus domestica 30CH produced clearly observable symptoms in healthy provers. On comparison, the proving remedy and the doctrine of signatures brought many parallels to light – the majority of which related to the mind, extremities and head. To gain a complete remedy picture of Malus domestica it is imperative that further research into the symptomatology of different homoeopathic potencies be conducted. / M

Thesis (MSc (Plant Pathology))--University of Stellenbosch, 2009. / ENGLISH ABSTRACT: Dry core rot (DCR) and wet core rot (WCR) are among some of the most important postharvest diseases of apples in South Africa. Mouldy core (MC) is also a symptom associated with the core region of apples, but it is not of economical importance since apple tissue surrounding the core region is not affected as is the case with DCR and WCR. The incidence of core rots in harvested fruits can be as high as 12%, but in general ranges from 3 to 8%. Infections and losses can also occur during fruit handling in pack houses and during storage. Additionally, yield losses also occur prior to harvest within orchards due to premature fruit drop of core rot affected fruits. The incidence of core rot diseases in apples differ among apple cultivars, with most Red Delicious varieties being susceptible to the development of core rots, whereas core rots have rarely been reported in other cultivars such as Granny Smith. The etiology and epidemiology of WCR and DCR are poorly understood. Although many fungal genera have been associated with the diseases, small-spored Alternaria species are mainly associated with DCR, whereas Penicillium species including P. roquefortii, P. expansum and P. funiculosum have mainly been associated with WCR. Dry core rot infections have long been known to occur pre-harvest, whereas WCR is primarily known as a post-harvest disease where infections take place during fruit handling in pack houses. Recently, Tarsonemus mites have also been indicated as being a potential role player in the etiology of core rot diseases. The mites have been hypothesised to carry pathogen spores into the core region of apples, and they may also possibly cause small wounds that facilitate pathogen entry. In South Africa, apple growers have recently reported WCR as being present prior to harvest, which has not been reported previously. Therefore, the first aim of the study was to investigate the incidence, as well as the causal agent/s of pre-harvest WCR. The incidence of WCR ranged from 0% to 1.7% in eleven orchards, and was in general lower than that of DCR (0.4% to 6%). Isolation studies from eight internal positions in WCR apples showed that Penicillium was the predominant fungal genus in most of the positions, including the lesion area. Morphological and molecular characterisation of Penicillium isolates from WCR showed that P. 2 ramulosum prov. nom. was the main species isolated from lesions, as well as other isolation positions. However, this species was also the main species isolated from DCR, MC and asymptomatic apples. Penicillium expansum was only isolated at low frequencies from WCR and DCR apples. Other Pencillium species that were occasionally isolated included P. glabrum, P. chloroloma, P. chermisinum and a putative new species with closest affinity to P. dendriticum (P. species aff. dendriticum) on a DNA nucleotide sequence basis. Pathogenicity and virulence studies using three different inoculation methods showed that P. expansum was the most virulent species, followed by P. species aff. dendriticum. The P. ramulosum prov. nom. isolates varied in their virulence, but were all considered to have low virulence. The role of Tarsonemus mites in the etiology and epidemiology of core rot diseases is poorly understood, and therefore the second aim of the study was to investigate some of these aspects. The specific aims of the study were to (1) investigate the ecology of Tarsonemus mites in Red Delicious and Granny Smith orchards during different apple developmental stages, (2) determine if there is a significant association of Tarsonemus mites with diseased (WCR and DCR) fruits and (3) determine if potential core rot pathogenic fungi are associated with the mites. Tarsonemus mites were found in all of the investigated apple developmental stages (buds, blossoms, 4cm diameter fruit, mature fruit and mummies), having the highest incidence in mummies and mature fruits from Red Delicious and Granny Smith orchards. In Red Delicious fruits the Tarsonemus mites were found within the core and/or calyx tube, whereas in Granny Smith fruits the mites were restricted to the calyx tube. In Red Delicious fruits there was a significant association between dry core rot as well as total core rot (wet- and dry-core rot) with the presence of mites in the core, as well as total mites (mites in core and calyx tubes). Fungal isolation studies from the Tarsonemus mites showed that they carried potential core rot fungal pathogens within the genera Penicillium and Alternaria. The Penicillium species isolated from the mites included two of the most virulent WCR species, P. expansum and P. species aff. dendriticum. / AFRIKAANSE OPSOMMING: Droë kernvrot and nat kernvrot is van die belangrikste na-oes siektes van appels in Suid- Afrika. Beskimmelde kern word ook met die kern van appels geassosieer, maar hierdie toestand is egter nie van ekonomiese belang nie, aangesien die weefsel rondom die kern nie geaffekteer word soos in die geval van nat- en droë kernvrot nie. Die voorkoms van kernvrot in vrugte na oes, kan vlakke van tot 12% bereik, maar oor die algemeen is die voorkoms tussen 3 en 8%. Infeksie en verliese kan ook voorkom gedurende die hantering en verpakking van vrugte in pakhuise en gedurende storing. Addisionele verliese in opbrengs kan ook voor-oes voorkom in boorde. Dit is te wyte aan voortydige vrugval van appels wat besmet is met kernvrot. Die voorkoms van kernvrot by appels verskil tussen kultivars. Meeste van die “Red Delicious” variëteite is vatbaar vir die ontwikkeling van kernvrot. Die toestand is egter skaars by ander kultivars soos Granny Smith. Die etiologie en epidemiologie van nat- en droë kernvrot word nie goed verstaan nie. ‘n Groot aantal swamgenera is al met kernvrot geassosieer. Klein-spoor Alternaria spesies word hoofsaaklik met droë kernvrot geassosieer en Penicillium spesies, insluitende P. roquefortii, P. expansum en P. funiculosum, word meestal met nat kernvrot geassosieer. Dit is lank reeds bekend dat droë kernvrot as voor-oes siekte kan voorkom, maar nat kernvrot is algemeen bekend as na-oes siekte waar infeksie tydens vrughantering en verpakking plaasvind. Daar is onlangs aangedui dat Tarsonemus myte potensiële rolspelers in die etiologie van kernvrot is. Hipoteties is die myte in staat om spore van die patogene in die kern van die appels in te dra, asook om klein wonde te veroorsaak wat infeksie deur patogene vergemaklik. In Suid-Afrika is nat kernvrot wat voor-oes in die boorde ontstaan onlangs deur boere aangemeld; hierdie toestand is nog nie op ‘n vorige geleentheid aangemeld nie. Die eerste doelwit van hierdie studie was dus om die voorkoms en veroorsakende organisme/s van voor-oes nat kernvrot te ondersoek. Die voorkoms van nat kernvrot was tussen 0 en 1.7% in elf boorde en was oor die algemeen laer as die voorkoms van droë kernvrot (0.4 tot 6%). Isolasiestudies uit agt interne posisies van nat kernvrot appels het getoon dat Penicillium die dominante swamgenus in die meeste posisies was, insluitend die letsels. Morfologiese en molekulêre karakterisering van 4 Penicillium isolate uit nat kernvrot letsels het aangedui dat P. ramulosum prov. nom. die spesie is wat die meeste geïsoleer is vanuit die letsels, asook ander isolasie posisies. Dié spesie was egter ook die mees algemene spesie wat uit nat- en droë kernvrot, asimptomatiese appels en appels wat slegs swamgroei in die kern gehad het, geïsoleer is. Penicillium expansum was ook in lae getalle uit nat- en droë kernvrotletsels geïsoleer. Ander Penicillium spesies wat ook soms geïsoleer is, sluit P. glabrum, P. chloroloma, P. chermisinum, asook ‘n moontlik nuwe spesie wat op DNA volgorde basis die naaste aan P. dendriticum (P. spesie aff. dendriticum) is. Studies wat patogenesiteit en virulensie van die isolate ondersoek het, is ook uitgevoer deur gebruik te maak van drie verskillende inokulasie metodes. Die studies het aangedui dat P. expansum die mees virulente spesie is, gevolg deur P. spesie aff. dendriticum. Die P. ramulosum prov. nom. isolate het variasie in virulensie getoon maar is oor die algemeen aanvaar om minder virulent te wees. Die rol van Tarsonemus myte in die etiologie en epidemiologie van kernvrot word nie goed verstaan nie en dus was die tweede doelwit van die studie om sommige van dié aspekte te ondersoek. Die spesifieke doelwitte was (1) om die ekologie van die Tarsonemus myte in “Red Delicious” en Granny Smith boorde tydens verskillende ontwikkelingstadiums van die appels te ondersoek, (2) om te bepaal of daar ‘n betekenisvolle assosiasie van Tarsonemus myte met siek (nat- en droë kernvrot) vrugte bestaan en (3) om te bepaal of potensiële kernvrot patogeniese swamme geassosieer is met die myte. Tarsonemus myte is gevind in al die ontwikkelingstadiums (knoppies, bloeisels, 4 sentimeter deursnee vrugte, volwasse vrugte en mummies) van appels wat ondersoek is. Die hoogste voorkoms van myte was in die mummies en volwasse vrugte van “Red Delicious”, asook Granny Smith kultivars gevind. In “Red Delicious” vrugte is myte in die kern en/of kaliksbuis gevind, maar in die Granny Smith vrugte was die myte tot die kaliksbuis beperk. In “Red Delicious” vrugte was daar ‘n betekenisvolle assosiasie tussen droë kernvrot, asook totale kernvrot (nat en droë kernvrot) met die teenwoordigheid van myte in die kern, asook totale myte (myte in die kern en kaliksbuis). Swam isolasiestudies vanaf die Tarsonemus myte het aangetoon dat potensiële kernvrot swampatogene in die genera Penicillium en Alternaria wel by die myte teenwoordig was. Die Penicillium spesies wat vanaf die myte geïsoleer is het twee van die mees virulente nat kernvrot spesies ingesluit, nl. P. expansum en P. spesie aff. dendriticum.

Thesis (MSc)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Apples are an important export commodity for the South African market, and postharvest losses that occur as a result of decay due to infection with pathogenic fungi such as Botrytis cinerea Pers., Penicillium expansum (Link) Thom. and Neofabraea alba (E.J. Guthrie) are of major concern for all parties concerned with fruit production and distribution. Decay control of these fungi is primarily managed through the use of synthetic fungicides; however, pathogen development of resistance to these fungicides and recent worldwide concern over healthier living and a greener environment has called for the discriminate use of synthetic chemicals. This has opened up an avenue for the development of safer and more environmentally friendly alternatives to control postharvest decays. The use of plant extracts and essential oils are favoured as natural sources of antimicrobials whilst still being safe for human consumption and having no negative impact on the environment. Allium sativum (garlic) is one such plant species that is well documented for its value in improving human health and is readily available for consumption not just as a flavour component of food but also to be taken as a daily herbal diet supplement. Given the antimicrobial effectiveness of garlic against human pathogens and ailments, its value as an antifungal agent against postharvest pathogens causing grey mould, blue mould and bull’s eye rot of apples was investigated in vitro and in vivo within this study. Furthermore, an attempt was made to elucidate the chemical components of garlic extracts by gas chromatography-mass spectrometry (GC-MS). All experiments in this study were carried out with garlic extracts prepared from fresh garlic bulbs. For the in vitro experiments, two extract preparations of garlic, one containing ethanol (Extract 1) and one where ethanol had been removed by evaporation (Extract 2), was tested for antifungal action within an amended media experimental design. Both extract preparations were each subjected to two dilution series (0-80% garlic extract) with water and ethanol as diluents. Both extract preparations were successful at retarding pathogen mycelial growth and spore germination; however, concentrations of Extract 2 (ethanol evaporated) and diluted with distilled water provided markedly better inhibition of B. cinerea and P. expansum than the ethanolic dilutions of extract 2. Both extract preparations yielded similar inhibitory results when tested against N. alba. Due to the results achieved in the amended media experiments, the use of a crude garlic extract without ethanol and diluted in water was considered to be the best option for further tests throughout the remainder of the study. In vitro volatile effects of crude garlic extracts at concentrations between 0 and 40% garlic extract were subsequently tested. Garlic volatiles were effective in inhibiting pathogen mycelial growth and spore germination of all three pathogens, at lower concentrations compared to the amended media experiments. In vitro volatile exposure with garlic extracts was more effective at inhibiting N. alba than direct application of the extracts. Curative and protective application of garlic extracts and clove oil for increased fungal inhibition through synergism was tested by direct and volatile exposure to the pathogens in vivo on three economically important apple cultivars; ‘Granny Smith’, ‘Golden Delicious’, and ‘Pink Lady’. Direct exposure of artificially wounded and inoculated fruit to the garlic extract and clove oil revealed that garlic extracts applied curatively but not protectively effectively controlled decay caused by B. cinerea and P. expansum on all apple cultivars. Both curative and protective applications were ineffective in controlling N. alba. In vivo volatile exposure to the garlic extracts and clove oil did not inhibit decay on any of the cultivars and was not effective against any of the three pathogens investigated. A full chemical profile analysis was done by GC-MS analysis of garlic extract samples. The compounds diallyl disulphide, allyl methyl trisulphide, allyl methyl disulphide and dimethyl trisulphide were detected in relatively high amounts. This result suggests that the abundance of sulphur and sulphur related compounds detected may be responsible for the antifungal action noted in the experimental studies. In conclusion, garlic was shown to have antifungal activity against B. cinerea, P. expansum and N. alba. The pathogens used in this study were not compared with each other, but undoubtedly each pathogens reacts differently to exposure to the garlic extracts. It would therefore be advisable to investigate the effects of the extracts on each of the pathogens in a more in-depth study. More investigations into the application of the garlic extracts is required before it may be recommended for use; however, results for the use of garlic extracts against these postharvest pathogens and the postharvest decay they cause are promising. / AFRIKAANSE OPSOMMING: Appels is ‘n belangrike uitvoerproduk vir die Suid-Afrikaanse vrugtebedryf, maar noemenswaardige na-oes verliese word weens bederf deur patogeniese swamme soos Botrytis cinerea Pers., Penicillium expansum (Link) Thom. en Neofabraea alba (E.J. Guthrie) ervaar. Dit raak alle partye betrokke met die produksie en verspreiding van hierdie vrugsoort. Hierdie swamme word hoofsaaklik met behulp van kunsmatige swamdoders beheer, alhoewel weerstand-ontwikkeling en wêreldwye bewusmaking van ‘n gesonder leefstyl en omgewing die gebruik van kunsmatige middels streng aanspreek en die ontwikkeling van veiliger en meer omgewingsvriendelike alternatiewe middels verlang. Plant-ekstrakte en essensiële olies kan dien as sulke middels en is natuurlike bronne van anti-mikrobiese aktiwiteit, is veilig vir menslike verbruik en het ook geen negatiewe invloed op die omgewing nie. Allium sativum (knoffel) is so ‘n plantspesie wat as alternatiewe middel gebruik kan word. Dit is bekend vir sy waarde in die verbetering van menslike gesondheid, is maklik bekombaar en word nie net as ‘n geurmiddel vir voedsel gebruik nie, maar ook as ‘n daaglikse krui-aanvulling. Gegewe die anti-mikrobiese doeltreffendheid van knoffel teenoor menslike patogene en kwale, is die werking (in vitro en in vivo) teen na-oes patogene wat grys skimmel, blou skimmel en teikenvrot in appels veroorsaak, in hierdie studie ondersoek. Bepaling van die chemiese samestelling van die knoffel-ekstrak is ook met behulp van gaschromatografie massa spektrometrie (GK-MS) onderneem.Vars knoffelbolle is vir elke eksperiment in hierdie studie gebruik met die voorbereiding van die knoffel-ekstrak. Vir die in vitro eksperiment is twee knoffel-ekstrakte voorberei, naamlik: ‘n ekstrak wat etanol bevat (Ekstrak 1) en een waarvan die etanol verwyder is met verdamping (Ekstrak 2). Die ekstrakte is getoets vir werking teen fungi in kultuur-medium.. Albei ekstrakte is verdun tot twee konsentrasie reekse (0-80%) met water en etanol as verdunningsmiddels. Albei ekstrakte het suksesvolle werking getoon teenoor die patogene ten opsigte van vertraging van miseliumgroei en spoor-ontkieming, alhoewel konsentrasies van Ekstrak 2, verdun met gesuiwerede water, patogene B. cinerea en P. expansum beter onderdruk het as Ekstrak 2 verdunnings met etanol.. Beide ekstrakte en hul afsonderlike verdunnings met etanol en water het soortgelyke resultate gelewer met onderdrukking van N. alba. Volgens resultate wat verkry is van die kultuur-medium eksperimente, is Ekstrak 2 verdun met gesuiwerde water beskou as die geskikste vir verdere toetse in hierdie studie. Die vlugtige effek van Ekstrak 2 is in vitro getoets by konsentrasies tussen 0 tot 40%. Die vlugtige stowwe van knoffel het al drie patogene se groei en spoor-ontkieming effektief onderdrukby laer konsentrasies as wat gebruik is in die kultuur-medium eksperiment. Dus is in vitro blootstelling van N. alba aan die vlugtige stowwe meer effektief as direkte toediening van die ekstrakte. Die voorkomende en beskermende effek van die knoffel-ekstrak, asook naeltjie-olie, is in vivo ondersoek om te bepaal of die stowwe saam sterker onderdrukking van die patogene kon bewerkstellig. Direkte en vlugtige blootstelling is op drie ekonomies-belangrike appel-kultivars getoets, naamlik: ‘Granny Smith’, ‘Golden Delicious’ en ‘Pink Lady’. Direkte blootstelling met die knoffel-ekstrak en naeltjie-olie aan gewonde en ge-inokuleerde vrugte het aangedui dat B. cinerea- en P. Expansum-bederf net beheer kon word indien knoffel voorkomend toegedien is vir al die ondersoekte appel-variëteite. Voorkomende en beskermende toediening was onsuksesvolle om N. alba te beheer. In vivo blootstelling van die drie patogene aan die knoffel-ekstrak en naeltjie-olie se vlugtige stowwe kon nie enige van die patogene effektief onderdruk nie en was onsuksesvol in bederf-beheer. ‘n Volledige chemiese profiel is saamgestel deur GK-MS ontleding van die knoffelekstrakte. Hoë vlakke van verbindings dialliel disulfied, alliel-metiel-tri-sulfied, alliel-metieldisulfied en dimetiel-trisulfied is bespeur. Die aantal vrye sulfied en sulfied-verwante verbindings in die ekstrak kan moontlik ‘n verduideliking bied vir die anti-swam werking waargeneem gedurende hierdie studie. Ten slotte: knoffel toon ‘n anti-swam werking teenoor B. cinerea, P. expansum en N. alba. Die patogene in hierdie studie is nie met mekaar vergelyk nie, omdat elkeen uniek en uiteenlopend op knoffel reageer het. Alhoewel die huidige studie alreeds belowende resultate gelewer het, moet die ekstrak se effek op elke patogeen onderskeidelik nog in diepte ondersoek word, asook die wyse van die toediening in die na-oes praktyk voordat hierdie middel aanbeveel kan word vir gebruik.