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.

South Africa is the second largest exporter of citrus fruit in the world. There has recently emerged a strong demand for lemons in the world market due to their nutritional value, culinary and non-culinary uses. During exportation, fruit are subjected to low temperature (-0.5°C) for varying periods of time as an obligatory quarantine treatment. However, lemons are sensitive to low temperatures and easily develop chilling injury during this obligatory quarantine treatment. This has become a major limitation to the expansion of South Africa’s lemon industry. Postharvest treatments with methyl jasmonate (MJ) and / or salicylic acid (SA) have been successfully used in horticultural crops to reduce chilling injury. A similar treatment was applied to ‘Eureka’ lemons. During the 2008 harvest season, postharvest fruit were either dipped in 10 or 50 μM MJ or 2 or 2.5 mM SA solutions. A control or no dip treatment was also applied. Three replicates of 15 fruits per treatment were used. During the 2009 harvest season the following postharvest treatments were applied as dips: 10 μM MJ, 2 mM SA, 10 μM MJ & 2 mM SA, 1 μM MJ & 0.2 mM SA, or 0.1 μM MJ & 0.02 mM SA solutions. A control or no dip treatment was also applied. Three replicates of 15 fruits per treatment were used. Subsequently fruit were stored at -0.5ºC for 0, 7, 14, 21, 28, 35, and 42 days, before being transferred to room temperature (25°C) for 7 days where after chilling injury was rated. Treatments with 10 μM MJ and / or 2 mM SA reduced chilling injury symptoms in lemons harvested during the 2009 season. Although no visual symptoms of chilling injury were observed during the 2008 harvest season, treatments with 10 μM MJ and / or 2 mM SA reduced fruit mass loss, delayed the occurrence of stress symptoms such as lipid peroxidation and suppressed accumulation of ROS in the rind. Treatments with 10 μM MJ and / or 2 mM SA were more effective in inducing antioxidant capacity and other defence compounds such as phenolics, ascorbic acid, carbohydrates and chilling injury responses such as accumulation of proline in the rind. This may have increased the chilling tolerance of fruit during the cold storage. Therefore, this study revealed that MJ and SA have the potential to reduce and delay symptoms of chilling injury in lemons. This lead to the suggestion that both, MJ and SA dips should be further tested as treatments to mitigate chilling injury in lemons. Future studies should focus more on preventing the injury itself or preventing the primary event of chilling injury. This could probably reduce the chances of secondary events to take place. / Thesis (M.Sc.Agric.)-University of KwaZulu-Natal, Pietermaritzburg, 2010.

Zinc and manganese have long been considered as essential micronutrients to plant growth, yet the interactions of the two nutrients on growth and development of plants have not been elucidated in their entirety. Silicon is not classed as an essential element, but has been found to improve growth of a number of crops, particularly of the Poaceae family. A simple water culture hydroponic system was developed to monitor the growth and development of a fruit crop (Cucumber – Cucumis sativus) under deficient, adequate and excessive applications of zinc and manganese. Plant growth parameters were monitored including leaf growth, plant height, plant fresh and dry mass, yield, fruit size and fruit mass. Nutrient uptake was also measured using inductively coupled plasma emission spectroscopy, whilst chlorophyll was determined spectrophotometrically. Plant nutrient analyses were also conducted using inductively coupled plasma emission spectroscopy. Silicon was found to have a beneficial effect on the growth of cucumbers and was incorporated as a treatment for this crop along with zinc and manganese since foliar silicon sprays were able to correct the occurrence of mineral deficiency symptoms. Along with plant growth measurements, nutrient uptake, plant nutrient analysis and chlorophyll determination, plant tissue was also analysed using transmission electron microscopy to establish the impact of silicon applications on the cell ultra-structure of cucumbers. Electron micrographs showed an increased presence of plasmodesmata in treatments excluding silicon. Such increased plasmodesmata connections under silicon deficient conditions could increase translocation of cell solutes due to reduced cell longevity. Results also confirmed the essentiality of zinc and manganese on plant growth and development as typical deficiency symptoms were observed. Typical toxicity symptoms were also recorded. Rates of uptake of nutrients corresponded with leaf growth and enlargement as well as yield. The chlorophyll concentration was not a clear indicator of nutrient application level. Typically, manganese and zinc interacted with iron, magnesium, calcium and potassium, affecting their uptake into the plant dependent on the level of manganese and zinc applied. Although non-essential, silicon improved plant growth, but had neither a relationship with the other nutrients evaluated nor affected the physical growth and development of the plants. Manganese and zinc, as essential to plant growth and development, affect the visual appearance of the plant as well as affect the plant biochemically due to their involvement in many growth and development processes. / Thesis (M.Sc.Agric.)-University of KwaZulu-Natal, Pietermaritzburg, 2010.

In South Africa, a shortage of litchi cultivars to expand the very short harvesting period has seriously threatened the viability of the local export orientated Litchi Industry. Only two cultivars have dominated commercial plantings for more than a century, namely ‘HLH Mauritius’ and ‘McLean’s Red’. The marketing period of litchis from South Africa coincides with large export volumes from competitor countries such as Madagascar, resulting in lower returns for local growers. To address this situation, the late season cultivar ‘Wai Chee’ was imported amongst others from Australia. ‘Wai Chee’ is harvested at a time in South Africa when there is a gap in worldwide litchi production, making the cultivar potentially very profitable. However, its potential and subsequent use in the industry is affected by small fruit size and questionable internal quality. As the South African Litchi Industry is export orientated and the qualities of ‘HLH Mauritius’ fruit currently dictate export requirements, solutions need to be found to improve fruit size and fruit quality in ‘Wai Chee’ litchi. In this study, a number of horticultural manipulation techniques were investigated with the aim of enhancing yield, fruit size and quality in ‘Wai Chee’ litchi. The use of chemical applications of foliar nutrients and plant growth regulators were found to improve certain fruit characteristics in ‘Wai Chee’. Foliar nutrient applications of nitrogen, potassium and calcium during the early stages of fruit set and -development improved fruit set and subsequently yield, and also increased fruit mass, fruit size and flesh mass. Treatments with potassium nitrate (KNOз), calcium nitrate (CaNOз) and calcium metalosate proved to be the most enhancing nutrient applications. Applications of synthetic auxins and auxin-like substances during the 2-3g stage of fruit development improved fruit size, fruit mass and flesh mass in ‘Wai Chee’. The combination treatment of Tipimon® (2,4,5-TP), applied at the 2-3g stage, followed by Maxim® (3,5,6-TPA) a week later, yielded the best results in this regard. With biological practices, pollination was found to have an influence on litchi tree- and fruit characteristics. Pollen source proved to have an influence on fruit set and fruit retention at harvest in ‘Wai Chee’. Initial fruit set was lower when using cross-pollination compared to the use of self-pollination in female flowers of ‘Wai Chee’, whereas final fruit retention was higher with the use of cross-pollinators when compared to retention of fruit with self-pollination. Although some beneficial effects with different pollen donors on fruit characteristics were observed, these effects were not significant, and would therefore necessitate further investigation. Pollen donor effect on quality parameters such as Titratable acid (TA)- and Total Soluble Solid (TSS) content of fruit was not significant. Cultivar differences regarding fruit characteristics and maturation rate were detected with the use of cultural practices such as bunch covering materials in ‘HLH Mauritius’ and ‘Wai Chee’. Beneficial effects on fruit size were obtained with thicker covering materials with nominal mass of 70 and 80 g/m² respectively on ‘HLH Mauritius’, while with ‘Wai Chee’, thinner covering materials (60 and 65 g/m²) showed enhancing effects. Maturation rate was significantly delayed only on ‘Wai Chee’ with the use of thicker covering materials (70 and 80 g/m²). Differences in colour were detected amongst different covering materials, but these should be verified with chromameter technology. Covering of fruit bunches for better fruit size and a later harvest date would, especially for ‘Wai Chee’ as a late season cultivar, be beneficial, since better prices are realised towards the end of the season. / Thesis (M.Sc.Agric.)-University of KwaZulu-Natal, Pietermaritzburg, 2010.

Clones of the Eucalyptus grandis x Eucayptus nitens (GN) hybrids were produced and selected through the CSIR‟s breeding programmes for colder plantation sites in South Africa. Some GN clones consistently exhibit high and superior pulp properties, which makes them valuable for commercial plantations in South Africa. In nurseries, stockplants are usually seven cm in length and maintained at high (100 x1.5 m-2) planting density. However, rooting frequency varies with season and little is known about the impact of position of cuttings on overall rooting frequency of a clone. The aim of this study was to investigate the effect of size and planting density of stockplants in mini-hedges, on the yield and subsequent rooting of cuttings from various positions of GN clones of known rooting potential (i.e. GN 018B: difficult-to-root and PP 2107: easy-to-root clones). Stockplants (10 cm vs. 20 cm) were established at high (100 x 1.5 m-2) and at low (25 x 1.5 m-2) densities for GN 018B and PP 2107 under commercial nursery conditions in a polyethylene tunnel. Cuttings were harvested every two to three weeks in September-October 2010 (spring), December 2010-January 2011 (summer), April-May 2011 (autumn) and June-July 2011 (winter). The harvested material was 5 – 7 cm in length and the light intensity received by individual stockplants at the two planting density levels was recorded. Harvested cuttings from the three positions (apical, middle and basal shoots) were used for: (i) rooting experiments under nursery conditions, (ii) bio-stimulant analysis using the mung bean bioassay, and (iii) analysis of soluble sugars. Between spring and summer 2010, the two GN clones established at low density yielded a similar number of cuttings, but differences in the rooting frequencies were significant in favour of PP 2107 clone. Similar observations were made at high density in terms of production of cuttings, but the significant differences in the rooting observations were reversed between the clones. The GN 018B clone had low rooting rates in summer under nursery conditions but its tissue extracts promoted higher rooting in the bioassay during that time, when compared to spring. Spring and summer had similar effects on rooting responses of PP 2107 cuttings in nursery and bioassay experiments. For both clones, short stockplants produced fewer cuttings but had a higher rooting frequency than cuttings from tall stockplants, with a high rooting frequency recorded from basal cuttings. Similar results were observed in the bioassay experiments which showed high rooting potential of mung bean hypocotyls cuttings using tissue extracts of PP 2107 cuttings maintained at high planting density. Although apical cutting tissues had high concentration of sugars (i.e. sucrose, glucose and fructose), their rooting rates were usually lower at high and low planting density compared to middle and basal cuttings. Sucrose concentration was the highest sugar present in stockplants grown under low planting density. A higher and lower rooting frequency was also observed in autumn although the two clones responded differently to Quambalaria eucalypti (Sporothrix eucalypti) disease infestations. Position, size and genotype had a significant impact on type and concentration of sugar (i.e. sucrose, glucose and fructose), particularly in PP 2107 clone, although rooting rates in the bioassay did not correlate with sugar contents of Eucalyptus cuttings. High carbohydrate (i.e. soluble sugar) content and auxin concentration increased production and subsequent rooting of cuttings across both clones, particularly in spring. Furthermore, rooting was enhanced by relatively higher light intensity intercepted by individual stockplants and in particular the GN 018B clone. Light intensity in the high and low planting densities caused variation in the rooting frequencies thereby increasing or decreasing soluble sugar and auxin concentrations of the two clones. Light intensity and fertiliser concentration received by tall and short stockplants impacted on endogenous hormone levels thereby increasing or decreasing rooting. High sugar concentration levels of PP 2107 clone increased its susceptibility to fungal infection thereby decreasing its rooting frequency in autumn, as its rooting rates increased in winter. Overall results of the investigation revealed that PP 2107 clone has higher rooting potential than GN 018B clone, in particular at high planting density and if stockplants are not infected by fungal diseases. Higher sugar levels were recorded in spring for PP 2107, although rooting rates of mung bean hypocotyl cuttings were higher in summer for GN 018B, suggesting that sugars have nothing to do with rooting of GN cuttings. Season, planting density and size of stockplants affect the rooting frequency of GN clone. Thus, short stockplants maintained at low and high planting densities are recommended for GN 018B and PP 2107 respectively, although the impacts of fertilisers and pathogen resistance on rooting rates still need to be investigated under similar conditions. / Thesis (M.Sc.Agric.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

Avocados are one of the major food sources in tropical and subtropical regions and are an important horticultural crop in South Africa. Avocados are exported over long distances and may have storage times of up to 30 or more days at temperatures of about 5.5oC. This procedure increases the risk of poor fruit quality, including physiological disorders, early softening and postharvest disease incidence. A major component of the postharvest diseases is Anthracnose caused by Colletotrichum gloeosporioides. Anthracnose infects unripe fruit and once infected, the fungus remains dormant in the fruit until ripening begins. This leads to a problem for producers and packers, as the presence of the disease cannot be detected on the pack line, and fruit is not removed. Anthracnose control is normally done through pre-harvest treatment with copper-based fungicides. While effective such treatment needs to be repeated frequently, resulting in copper residues on the avocados. The study was conducted to investigate the effects of phosphoric acid and potassium silicate on known antifungal compounds and critical enzymes of the pathways elemental for systemic resistance inducers, so as to evaluate the potential for using them as alternatives to or in conjunction with, copper fungicides in the control of Anthracnose in avocado fruit. The study included storage temperature and time variations, to take account of the logistics in shipping avocado fruit to distant markets. Pre- and postharvest applications of phosphoric acid and potassium silicate were used, and after harvest, fruit were either ripened at room temperature (22oC) without storage or stored for 28 days at temperatures of 5.5oC or 2oC before analysis. Concentrations of phenolics, activity of the enzyme phenylalanine ammonia lyase (PAL) and a known antifungal diene were determined in the fruit exocarp. Pre-harvest treatments of phosphoric acid showed that the highest phenolic concentration was found in fruit harvested 14 days after application for fruit stored at room temperature. For fruit stored at 5.5°C it was seen that as fruit softened, phenolic concentrations increased compared with hard fruit immediately after storage, with the highest increase noted for fruit harvested 7 days after application. When comparing the three storage temperatures, phenolic concentrations were enhanced most when fruit was stored at 2°C. Postharvest treatments showed a significant increase in phenolic concentrations for potassium silicate treated fruit stored at room temperature and 2°C when determined immediately after storage. Fruit stored at 5.5°C showed an increase in phenolic concentrations as it became softer. When considering PAL enzyme activity, it was found that postharvest treatments of both potassium silicate and phosphoric acid influenced enzyme activity, with potassium silicate having greater effects. Similarly, an increase in PAL activity was noted in the pre-harvest phosphoric acid treatment harvested 14 days after application for fruit ripened immediately as well as fruit stored at 5.5°C. Fruit stored at 2°C showed the highest PAL activity for fruit harvested 7 days after application. No results were obtained in the analysis of antifungal compounds for both pre- and postharvest treatments. However, it is suggested that the antifungal diene could follow similar trends to those found for phenolics. It is concluded that applications of both phosphoric acid and potassium silicate do create changes in phenolic concentrations and the activity of the enzyme PAL which is involved in the synthesis of phenolic compounds known to possess antifungal properties. It is therefore possible that phosphoric acid and potassium silicate may be used as part of an integrated programme for Anthracnose control, and should be tested as potential alternatives for high volume copper-based fungicides. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

The susceptibility of the 'Pinkerton' avocado cultivar to mesocarp discolouration, after storage, has seriously threatened its export from South Africa. This disorder has proven to be complex, requiring a better understanding of the fruit's physiology. The purposes of this study were to identify the role of pre- and postharvest factors, or their interactions, in the development of the problem. This was done by obtaining fruit from several production areas of varying mesocarp discolouration histories (referred to as "high", "medium" or "Iow risk" areas) during the 2000 and 2001 seasons. Fruit were stored at 8, 5.5 and 2°C for 30 days, as well as ambient (20°C). Evaluations of fruit quality were made before and after storage, as well as after softening. Once removed from storage the weight loss (during storage) was determined, and fruit firmness and carbon dioxide (C02) production rates monitored daily. It was found that temperatures below the recommended shipping temperature of 5.5°C, i.e. 2°C, produced the best internal fruit quality. This was supported by the membrane integrity studies that showed less membrane stability at the warmer storage temperature of 8°C. Furthermore, remained hard during storage and subsequently had an extended shelf life. Fruit origin was also found to play a major role in browning potential, with discolouration being consistently more severe in fruit from "high risk" areas and increasing in severity as the season progressed. The rate of CO2 production was found to follow a similar trend, with rates increasing as the season progressed, and also being slightly higher in fruit from "high risk" areas. The higher CO2 production rates were thought to be related to a decrease in membrane integrity as the season progressed. While storage temperature was not found to have a significant effect on the rate of CO2 production after storage, it did affect the time taken to reach the maximum rate, with fruit stored at 2°C taking longer. Biochemical analyses to determine the concentration of total phenolics and the activity of the enzyme polyphenol oxidase (PPO) also showed that the potential for browning was initiated by preharvest conditions. While no significant differences were found between growers with regards to total phenol concentrations, the PPO activity was found to be higher in fruit from poor quality areas, and subsequently browning potential was expected to be higher in these fruit. It was, however, found that the potential for browning could be reduced by storing fruit at 2°C, as this decreased the total phenolics concentration. This evidence further emphasized the idea that storage at 2°C could be highly advantageous. Fruit mineral analysis showed that certain key elements played a significant role in the severity of mesocarp discolouration, with excessive fruit nitrogen and decreasing copper and manganese concentrations appearing to play major roles. The high fruit nitrogen concentrations were suspected to reflect fruit grown on very vigorous trees, resulting in shoots competing with fruit for available reserves. It is suggested that 'Pinkerton' of a quality acceptable to the market, can be produced by manipulating source:sink relationships, particularly through decreasing the availability of nitrogen, followed by low temperature (24° C) shipping. Future work should concentrate on manipulation of source:sink relationships, to take account of both climatic conditions and leaf to fruit ratios. The evaluation of chlorophyll fluorescence as a tool for predicting mesocarp discolouration potential in 'Pinkerton' proved to be unsuccessful in this study and future studies may require modifications to the current technique. It is suspected that differences in chlorophyll content, for example, between fruit from different origins, will have to be taken into account when interpreting results. The success of using 2°C storage to improve the internal quality on 'Pinkerton' fruit prompted further studies, during 2004, to ensure that the development of external chilling injury would not decrease the marketability of the cultivar. Low temperature conditioning treatments, prior to storage, proved to be highly successful in reducing the development of external chilling injury, thus further improving fruit quality as a whole. Preconditioning treatments consisted of fruit that were kept at either 10°C, 15°C or 20°C for 1 or 2 days before being placed into storage for 30 days at 2°C or 5.5°C. All preconditioning treatments were compared to fruit that were placed directly into storage. The effect of fruit packaging on moisture loss (as determined by weight loss) and chilling injury was also investigated using unwaxed fruit, commercially waxed and unwaxed fruit individually sealed in micro-perforated polypropylene bags with an anti-mist coating on the inside (polybags). Holding 'Pinkerton' fruit, regardless of packaging treatment, at 10°C for 2 days prior to storage at 2°C or 5.5°C significantly decreased the severity of external chilling injury. The use of polybags during preconditioning and storage showed potential in further reducing the development of external chilling injury, although the higher incidence of fungal infections in these fruit needs to be addressed. The determination of proline concentrations in fruit exocarp tissue after storage was helpful in determining the level of stress experienced by fruit that were subjected to different packaging and preconditioning treatments. In this study waxed fruit subjected to 1 d preconditioning at 10°C, 15°C or 20°C or placed directly into storage at 2°C showed very high proline concentrations and also displayed more severe external chilling injury, despite unwaxed fruit losing more weight during these treatments. The role of moisture loss thus needs further investigation. The thickness and method of wax application was thought to play an important role in the higher external chilling injury ratings in this study as waxed fruit often developed chilling injury symptoms around the lenticels and it was suspected that either the lenticels were damaged by the brushes used to apply the wax or that the lenticels became clogged thus resulting in reduced gaseous exchange. Nevertheless, the success of low temperature conditioning in reducing external chilling injury, while maintaining sound internal quality, may enable storage temperatures to be dropped even further, thus enabling South Africa to export avocados to countries that require a cold disinfestation period prior to entry to eliminate quarantine pests (e.g. fruit fly). / Thesis (Ph.D.Agric.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

New emerging markets such as Japan and the United States require cold sterilisation of South African citrus fruit as a phytosanitary standard against fruit fly. However, citrus fruit are chilling susceptible, with lemons being the second-most chilling susceptible after grapefruit. Chilling injury is a physiological rind disorder; the occurrence of which is despite its prevalence in horticultural commodities, not well understood. Therefore, the aim of this study was to investigate physiological compounds regulating chilling susceptibility or resistance in citrus fruit, with special emphasis on lemons. Furthermore, the potential of hot water dips or “molybdenum soaks” to maintain a certain level of physiological compounds which determine manifestation of chilling injury symptoms in citrus fruit was investigated. Moreover, it was attempted to create an understanding of the order in which physiological compounds mitigate chilling injury. Lemon fruit from different farms known to be chilling susceptible or resistant were obtained during the 2007 and 2008 harvest season. Thereafter, fruit were treated by soaking for 30 min in 1μM NaMo04.2H20 solution followed by a 2 min HWD 47 or 53°C. Treated fruit were waxed, weighed and stored at -0.5°C for up to 28 days and sampled for chilling injury evaluation 7, 14, 21, or 28 days into cold storage. A second evaluation was carried out five days after withdrawal from cold storage to allow development of chilling injury symptoms as a shelf-life simulation. After the second evaluation fruit were peeled, peel freeze-dried, milled using mortar and pestle and stored at -21°C for further physiological analysis. Freeze-dried peel was analysed for soluble sugars (glucose, fructose, sucrose), vitamin C (ascorbic acid), vitamin E (α-tocopherol), β-carotene, polyamines (putrescine, spermine, spermidine), specific flavanones (naringin and hesperidin) using HPLC-UV-Vis detector and proline, total antioxidant assays (FRAP, ABTS, DPPH), total phenolics, total flavonoids, lipid peroxidation using spectrophotometry, as well as for the heat shock protein (HSP70) using electrophoresis and silver-staining. Chilling susceptibility of lemon fruit varied with fruit source; those sourced from Ukulinga and Eston Estates were chilling resistant, while fruit from Sun Valley Estates showed chilling injury symptoms after 28 days of cold storage plus five days shelf-life. Furthermore, hot water dips (HW) 53°C, 1 μM Molybdenum (Mo) and 10 μM Mo plus HW 53°C significantly reduced chilling injury symptoms compared with the control and HW 47°C. In addition, Sun Valley Estates fruit also showed higher fruit weight loss compared with non-chilling resistant lemons. The alignment of higher fruit weight loss during storage with chilling susceptibility ascertains the use of weight loss as a non-destructive parameter for chilling susceptibility. With respect to flavedo sugars, glucose was found to be the dominant soluble sugar with multi-functional roles during cold storage. This plays a significant role in mitigating cellular stress. Chilling susceptible lemons from Sun Valley Estates had low flavedo glucose concentrations and, therefore, little conversion of glucose to ascorbic acid was possible resulting in a low antioxidant capacity. However, treatments with HW 53°C and Mo soaks seemed to enhance the enzymatic conversion of glucose to ascorbic acid leading to a higher antioxidant capacity in the flavedo of such treated fruit. Furthermore, glucose also feeds into the pentose phosphate pathway which is coupled with the shikimate pathway synthesizing secondary metabolites, especially of the phenolics group. The decrease in glucose was aligned to the levels of total phenolics, but not to that of β-carotene, naringin and hesperidin through 28 days into cold storage period. Moreover, as glucose also feeds into shikimate pathway, simultaneously an increase in proline flavedo concentration was observed. Proline is an antioxidant synthesized from glutamate; as cellular glucose decreases so does the total antioxidant capacity during cold storage. Ascorbic acid is a dominant and potent antioxidant in lemon flavedo as proven with the FRAP, ABTS and DPPH assays. Chilling resistant fruit have significantly higher ascorbic acid conversion. Furthermore, ascorbic acid also acts to generate the α-tocopheroxy radical to further important membrane-bound antioxidant, vitamin E (α-tocopherol equivalent). Furthermore, the DPPH assay was found to be effective in quantifying total antioxidants in lemon flavedo since it detects both lipophilic and hydrophilic antioxidants compared with the ABTS and FRAP assays which are bias to the estimation of liphophilic or hydrophilic antioxidants, respectively. The hot water and molybdenum treatments increased total antioxidants (DPPH assay) with reduced lipid peroxidation 7 days into cold storage and therefore, reduced chilling symptoms in fruit from Sun Valley Estates. The capacity of antioxidant to scavenge reactive oxygen species (ROS) was increased during cold storage and membrane stability significantly improved. Furthermore, putrescine as low valency polyamine was reduced as such compound acted as precursor to the synthesis of the high valency polyamines, spermine and spermidine. Chilling susceptible lemons from Sun Valley Estates showed increased soluble-conjugated polyamines as a response to stress. Furthermore, HW 53°C, 1 μM Mo and 10 μM Mo plus HW 53°C significantly increased the protein concentration and, therefore, likely also the occurrence of proteins with 70kDa (as estimator of HSP70). Additionally, the concentration of conjugated high valency polyamines was also increased, resulting in reduced chilling injury symptoms. The effect of ROS has only been viewed as damaging, while recently their role has also been viewed as stress acclamatory signalling compounds when produced concentrations below critical damaging threshold. Therefore, hot water dips seems to signals synthesis of total protein which include HSPs which then act throughout cold stress to protect other protein and channel other damaged proteins towards proteolysis. While molybdenum increased ROS production below damaging critical threshold, with ROS signalling stress acclimation by further signalling production of bioactive compound with antioxidant properties. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

The South African Citrus Industry is the second largest exporter of citrus, after Spain. The industry is under pressure to supply high quality fruit as well as to expand into new, high paying markets. However, high paying markets such as Japan and the USA require cold sterilised fruit as obligatory quarantine treatments against Mediterranean fruit fly (Ceratitis capitata) in order to reduce any possible spread of the pest. Citrus fruit originated from tropical climates and hence are chilling susceptible. Chilling injury symptoms appear as dark brown spots, pitting and/or decay when fruit are transferred to shelf temperatures; thus reducing the marketability of citrus fruit. Therefore, there is need for methods to mitigate chilling injury. Previous studies have shown silicon to mitigate many forms of stress without any hazardous effect on human health. Thus, the aim of the study was to investigate the potential of post-harvest silicon dips in mitigating chilling symptoms in citrus fruit. Briefly, fruit from two sources (Ukulinga Research Farm and Ithala Farm) were dipped in different silicon concentrations (0, 50, 150, and 250 mg ℓ-1) for 30 minutes and thereafter stored at -0.5 or 2⁰C for up to 28 days with weekly evaluation for chilling injury symptoms. Total antioxidants were determined using FRAP, ABTS, and DPPH assays under spectrophotometer. In addition, sugars, ascorbic acid, phenolics and flavonoids were analysed using High Performance Liquid Chromatography (HPLC). Fruit from Ukulinga Research Farm showed significantly higher total antioxidants (ascorbic acid total phenolics and specific flavonoids hesperidin and naringin) and sugars relative to fruit from Ithala Farm. Low concentrations of silicon dips significantly reduced the appearance of chilling injury symptoms by inducing an enzymatic conversion of glucose to ascorbic acid, thereby increasing the antioxidant capacity of chilling susceptible fruit. Moreover, silicon increased the concentration of total antioxidants, total phenolics and total flavonoids. High silicon concentrations had a negative effect on post-harvest quality of lemons by increasing fruit weight loss and electrolyte leakage, resulting in appearance of chilling symptoms. In conclusion, the study showed that silicon had potential to reduce chilling injury. However, high silicon concentrations raised concern, in particularly, on fruit appearance. / Thesis (M.Sc.Agric.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

There is at present a growing concern and awareness of the endangered status of many indigenous South African plants in the wild, a number of which have potential for commercial production. One such example is Gladiolus scabridus, a vulnerable species endemic to the mountains of northern KwaZulu-Natal and southern Swaziland. It has considerable potential due to its floral characteristics. However, little is known about its horticultural requirements. Thus propagation and cultural practices were investigated with the aim of both conservation and commercialisation of the species. The ideal conditions for G. scabridus seed germination were determined. The presence or absence of light had no significant effect. Optimum germination was achieved at 20 QC of both winged and dewinged seeds. Higher temperatures appear to have a negative effect on germination and none of the winged seeds germinated at 30 QC. Under ideal conditions, fresh seed showed significantly higher vigour and viability than stored seed although the germination of stored seed was hampered by a higher internal fungal content. Successful tissue culture protocol was established for G. scabridus axillary bud and cormel halve explants. During the initial shoot initiation and proliferation stages (stage I and 11), the significantly higher shoot numbers occurred in the absence of growth regulators in both explant types. However, shoots with 1.0 mg 1 -1 6-benzyl-amino-purine (BAP) and 0.5 mg 1 -1 1-naphthalene-acetic-acid (NAA) were healthier in appearance. Higher levels of 5.0 mg 1 -1 NAA inhibited shoot production and encouraged root development in cormel halve explants. During stage 11, axillary bud explants showed root and cormlet development. More roots were initiated without growth regulators, whilst 5.0 mg 1 -1 NAA resulted in significantly better cormlet development. Shoot and cormlet growth of cormel halve explants during stage II was not significantly affected by the presence or absence of NAA and BAP. Significantly more roots were produced with 5.0 mg 1 -1 NAA. During stage III (rooting), the presence of activated charcoal (AC) was essential for the initiation and development of roots in vitro. Root and cormlet development in all explant types was significantly affected by the interaction between the previous treatments from stages I and 11 and the new treatments. During stage IV (hardening off) , most explants died down in the hardening off media leaving resting cormlets. There was a significant interaction between 1.0 mg 1 -1 NAA and 0.3 % AC from stage III, resulting in significantly more cormlets in both axillary bud and cormel halve explants. There was successful cormlet growth after cold storage which is advantageous in reducing the need for acclimatization. G. scabridus corms were successfully forced out of their normal flowering period. Temperatures of 10,5.5 and 2 QC showed successful corm initiation although the corms need to be stored for longer at warmer temperatures. It is suggested that a treatment of 2 QC for 6 weeks is the optimal condition for forcing G. scabridus corms. Although originating from a stressful environment, G. scabridus appears to show capacities for improved growth under controlled conditions. Fertilizer applications enhanced growth and reduced the time to flowering. Nitrogen (N) was found to be important for vegetative growth, flowering and daughter corm development, whereas potassium (K) influenced cormel production. Fertilizer with higher N and lower K is appropriate for the beginning and middle of the growing season and then adjusted to a lower N and higher K fertilizer to promote cormel formation. G. scabridus was found to produce prolific numbers of cormels which is an important source of plant material. Mineral leaf analysis showed that optimum levels for wild species are lower than those for hybrid gladioli with the optimum levels affected by physiological corm maturity and subsequent plant growth. Norms for postharvest handling of cut G. scabridus spikes have been developed. Spikes held in 2 % sucrose had a longer vase life and better floret opening and quality than those kept in distilled water, Prolong, Chrysal, 2 % sucrose and 2 % ethanol, 1 % sucrose, 4 % sucrose, 1 % sucrose and 0.5 % JIK, 2 % sucrose and 0.5 % JIK and 4 % sucrose and 0.5 % JIK, 2 % fructose and 2 % glucose. The use of commercially available solutions should be used with caution. Florets produced a climacteric-like CO2 peak, but levels of ethylene were unmeasurable. Cold storage and the use of polypropylene sleeves delays senescence. G. scabridus spikes secrete droplets of a sticky substance which was confirmed to be extra floral nectar through HPLC analysis. Market research revealed a positive response to the species from consumers and retailers alike with potential for cultivation as a cut flower and bedding plant. However, the cost will determine supply and demand. A field study conducted at Bivane Dam, northern KwaZulu-Natal, confirmed that G. scabridus colonies prevail in rocky, quartzite outcrops where they become wedged between the rocks. Plants were found at different stages of development with populations of not more than 108 plants per colony. Soil data of G. scabridus sites was compared to that of two sites nearby. It was found that G. scabridus soils are higher in phosphorus (P), zinc (Zn) and organic carbon. Leaf analysis confirmed that they have adapted their growth to low nutrient levels. The G. scabridus studies have clearly shown that the species can be successfully moved from a wild plant to a commercially viable one and in so doing its conservation status can also be improved. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.