Horticultural Science
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Item Development of a sulphur free litchi storage protocol using sealed polypropylene bags.(2006) Archibald, Alison Joy.; Bower, John Patrick.; Bertling, Isa.The use of sulphur as a method of postharvest disease control and colour retention in litchis is soon to be restricted by the European Union. It is therefore essential that new postharvest treatments and packaging techniques be developed in order to retain internal and external fruit qualities and thus allow for export. Good litchi quality is not only important for the export market but also for use on the local market. In this study, alternative methods for postharvest quality control were investigated with the aim of extending the litchi storage life to 40 days under modified storage. Packaging the fruit in polypropylene bags significantly decreased fruit water loss and resulted in an increase in shelf life, as determined by red colour and overall rind appearance. There was no distinct advantage of amodified atmosphere. The use of a punnet, lined with absorbent sheeting and placed within the sealed polypropylene bag, further improved the shelf life. The absorbent sheeting reduced the amount of free water and resulted in little pathogen infection, while the punnet was effective in protecting the fruit from damage. It was notable that most water loss occurred within the first 10 days of storage and that the majority would actually take place during the cooling phase. A hydrocooling technique was therefore investigated and was found to not significantly decrease water loss, possibly due to not hydrocooling the fruit for a long enough period of time. Temperature management was extremely important for both colour retention and pathogen control. It was found that treatments stored at 5.5QC showed better colour retention after the 40 days storage than the 1QC storage treatment. The higher storage temperature, however, enhances the potential for postharvest diseases. Three compounds, namely ISR 2000, 'Biosave' and F10, were tested for pathogen control. 'Biosave' showed the best results with the most effective concentration being 100 mill water and good pathogen control occurred when storage was at 10 C. Polyphenol oxidase (PPO) activity in the litchi rind was evaluated as it is thought to be closely related to browning of litchi fruit, probably due to the degradation of phenolics by PPO. Brown fruit had a high PPO activity whilst red fruit had much lower activity. It was also shown that PPO activity decrease over storage time, possibly due to product inhibition of the enzyme. The internal quality of the fruit was determined using the T88: acid ratio of the pulp, as it is well correlated to mean eating quality. For fruit to have excellent taste, it must have a T88: acid ratio of between 31:1 and 60:1. All the fruit had a ratio that met this criterion and would therefore ensure good eating quality.Item Gladiolus scabridus - the road to conservation and commercialisation.(2005) Campbell, Tracey Barbara.; Bower, John Patrick.; Van Niekerk, Renate.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.Item Factors affecting mesocarp discolouration severity in 'Pinkerton' avocados (Persea americana MILL.)(2005) Van Rooyen, Zelda.; Bower, John Patrick.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).Item A preliminary study on the effect of climatic conditions and fruit mineral concentration on the development of lenticel damage in 'Tommy Atkins' and 'Keitt' mangos (Mangifera indica L.) and rind pitting in 'Benny Valencia' oranges (Citrus sinensis).(2008) Magwaza, Lembe Samukelo.; Bower, John Patrick.; Kruger, Frans J.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.Item Investigation into the feasibility of a dairy processing enterprise through action research with a group of women from an LRAD project.(2007) Mans, Susanna Elizabeth.; Hendriks, Sheryl Lee.Poverty is caused by inadequate income. Effective assistance to small-scale enterprises may benefit the urban and rural poor. Small enterprises have the potential to create employment, reduce poverty and support sustainable livelihoods, as small enterprises satisfy local needs, provide employment opportunities in local areas, increase income, ensure food security and improve health. The Department of Land Affairs has recently introduced policies such the Land Reform Policy to correct the injustices of South Africa’s past. During 2005, the KwaZulu-Natal Department of Agriculture and Environmental Affairs introduced the Agrarian Revolution Programme, in an effort to close the gap between the first and second economy. The Agrarian Revolution and the LRAD programmes encourage agribusinesses through the formation of co-operatives or trusts. The purpose of this study was to investigate the potential of an envisaged processing enterprise with a group of women from an LRAD project, within the Engadini Community Trust. An action research methodology was followed. A training manual was developed and training conducted with the Engadini women’s group through which they prepared a business plan, developed prototype products; conducted market research; estimated prices for the products and developed a marketing plan. The study results indicate that the group would have the technical skills to make a good quality product. The group is likely to face problems ensuring a sustainable supply of raw materials as the current supply of milk would hardly be sufficient to supply the target market sampled during the consumer survey. The success of the dairy processing enterprises depends heavily on the ability of the Engadini Trust to overcome internal social problems. Conflict between the Engadini dairy processing enterprise members and conflict within the larger Engadini Community Trust had a crippling effect on the dairy processing enterprise. The main recommendations for the group are that the group needs to secure a sustainable supply or fresh milk. A dairy processing facility that will meet food safety standards needs to be erected for the enterprise. The location of the dairy processing enterprise needs to be re-considered. To overcome internal conflict within the group, the women’s group needs to gain representation on the Engadini Community Trust’s Executive Committee, to ensure that the dairy processing enterprise interests are considered by the Trust. The main recommendations for small-scale enterprises are that they should ensure to have sustainable supply of good-quality raw materials and a reliable, sustainable market, even if the market is relatively small. Policy recommendations are that Government Departments such as the KwaZulu-Natal Departments of Agriculture and Environmental Affairs and Economic Development should consider adapting policies that will focus on funding individual enterprises, rather than group enterprises. A directorate or sub-directorate dedicated to the development of small and medium sized food processing enterprises should be created. The marketing section of the KwaZulu- Natal Department of Agriculture and Environmental Affairs should provide marketing information for small-scale food processing enterprises available to extension officers and small-scale enterprises.Item Optimisation of propagation methods in Prunus persica (L.) Batsch.(2001) Lebese, Thabiso C.; Bertling, Isa.; Bower, John Patrick.Propagating methods for peach (Prumis persica L. Batsch.) are currently limited to the use of seeds or cuttings. Most of the rootstocks commonly used for establishing peach trees commercially originate from a narrow genetic base. The most commonly used peach rootstock in South Africa is 'Kakamas', which has disadvantages such as slow growth, and a high susceptibility to certain pests and diseases. 'Kakamas' is classified as a mid to late ripening cultivar, hence, its use as a rootstock is restricted in early ripening cultivars. Optimising peach seedling and cutting production may on the other hand increase more selection material that will increase genetic variability and also serve as the basis for future production and selection of rootstocks for peach nursery establishments. Hence, this study was designed to improves success of peach cutting production by increasing the rooting percentage and the survival rate of cuttings. This was firstly achieved by the application of various concentrations of IBA and several rooting-cofactors. Secondly, rooting success of cuttings taken at different seasons was studied. The response of two cutting positions to various IBA and rooting-cofactor was evaluated. Attempts were also made to overcome the requirement of peaches for a seed stratification period by using plant growth regulators. Studies to optimise the establishment and growth of embryos of early to late ripening cultivars ('Klara', 'Oom Sarel' and' Summer Giant') in vitro were undertaken to overcome the problem of small underdeveloped embryos which are often obtained from early ripening peach cultivars. The early ripening peach cultivars 'DeWet', 'Earlibelle' and 'Florida Prince' were propagated by softwood, semi-hardwood and hardwood cuttings prepared from August 1999 to July 2000. It was found that softwood cuttings prepared in October and November responded well to an IBA application of 1000 mg 1(-1), resulting in 100 % rooting in all three cultivars. In 'Florida Prince' and 'DeWet' the rooting-cofactors chlorogenic acid and phloroglucinol at the concentration of 100ug 1(-1) yielded 77 % and 81 % rooting, respectively while quercitin and rutinin gave only 46 and 44 % rooting for all cultivars. The basal portion of the cutting rooted better than the terminal portion (78 % versus 58 %). Rooting percentage differed in all treatments in response to IBA application and rooting-cofactors 74 % for 'Florida Prince', 62 % for 'DeWet' and 54 % for 'Earlibelle'. The use of IBA and Ca-EDTA proved to be beneficial for rooting of 'Florida Prince' cuttings and resulted in a rooting percentage of 86 % in wounded hardwood cuttings of this cultivar. It was also shown that the simple sugars glucose, fructose and sucrose as well as the sugar alcohols sorbitol and mannitol accumulated at the base of the cutting during adventitious base root formation if the cutting bases were treated with 1000 mg 1(-1) IBA. The concentration of these sugars and sugar alcohols were lower in untreated cuttings compared to IBA treated cuttings during the adventitious root formation process. Studies on the origin of adventitious root formation in stem cuttings of peaches were conducted using light microscopy and transmission electron microscopy. These studies revealed that adventitious roots originate (in peaches) in the vicinity of the vascular bundle tissue and in the cells around them. It was found, however, that adventitious root formation in IBA treated cuttings is associated with the formation of root primordia. These may trigger root initiation and ultimately the development of adventitious roots. In embryo culture studies the highest number of roots and greatest length of roots per embryo as well as the highest number of embryos forming roots was achieved when either the medium of Murashige and Skoog (MS) (1962) or of Steward and Hsu (SH) (1978) were employed. The Woody Plant medium (Lloyd and McCowan, 1978) and the medium after Schenk and Hildebrandt (1972) were found to be less effective when compared to MS and SH media. Addition of GA(3) (0.01 mg 1(-1)), BAP (0.2 mg 1(-1)) or GA(3) (0.01 mg 1(-1))+ BAP (0.2 mg 1(-1))+IBA (0.5 mg 1(-1)) to the media gave the best results with respect to embryos forming roots (86 %), number of roots per embryo (8.0) and total length of roots per embryo (7.57 cm). Furthermore, the following conditions for optimal rooting of peach embryos were established: pH 5.2, 1.5 g 1(-1) agar, 60 g 1(-1) sucrose and 16h light/8h darkness photo period. Further studies on the influence of the stratification temperature on germination of embryos revealed the highest germination percentage after exposure to 40 C ±2 constantly for 30 to 60 days. However, this chilling requirement can be successfully substituted by the addition of GA(3) , kinetin, zeatin, BA and thiourea to the culture media.Item Effect of postharvest silicon application on 'hass' avocado (Persea americana Mill.) fruit quality.(2010) Kaluwa, Kamukota.; Bertling, Isa.The South African avocado industry is export-orientated with forty percent of total production sold overseas. The avocado fruit is a highly perishable product with a relatively high rate of respiration which results in the quick deterioration of fruit quality. Good phytosanitary procedures are a necessity in ensuring good product quality. Due to the threat of pests and diseases becoming resistant to the conventional chemicals currently used to control them, there has been a great need to diversify from their usage. Silicon (Si), being the second most abundant element (28%) in the earth’s crust after oxygen, is a major constituent of many soils and has been associated with disease resistance in plants for a long time. It has been used in a number of crop species to provide resistance against pathogenic agents. In some horticultural crops Si has been found to offer protection against fungal infections by strengthening cell walls, thus making it more difficult for the fungi to penetrate and colonize the plant. The aim of this research was to investigate the effects of postharvest silicon application on the quality of ‘Hass’ avocado fruit. The specific objectives included investigating the effect of silicon on the ripening pattern as well as the metabolic physiology of the avocado fruit. Avocado fruit were obtained from two locations in the KZN Midlands (Everdon Estate in Howick and Cooling Estate in Wartburg). Fruit were treated with different forms of Si (potassium silicate (KSil), calcium silicate (CaSil), sodium silicate (NaSil) and Nontox-silica® (NTS)) at concentrations ranging from 160 ppm to 2940 ppm. After dipping for 30 minutes in the silicon treatments, the fruit were stored at -0.5°C, 1°C, 5°C or at room temperature (25°C). Energy dispersive x-ray (EDAX) analysis was then conducted on the exocarp and mesocarp tissues to determine the extent of silicon infiltration within each treatment. Firmness measurements, ethylene evolution and CO2 production were recorded as fruit approached ripening. The CO2 production of fruit that were stored at room temperature was analysed daily until they had fully ripened, while fruit from cold storage were removed weekly to measure respiration. Mesocarp tissue from each fruit was extracted using a cork borer and subsequently freeze-dried and stored for physiological analysis. The freeze-dried mesocarp tissue was then finely ground and later analysed for sugar content, total anti-oxidant capacity (TAOC), total phenolic (TP) content and phenylalanine ammonia lyase (PAL) activity using their respective assays. Statistical analyses were carried out using GenStat® version 11 ANOVA. Treatment and storage temperature means were separated using least significant differences (LSD) at 5% (P = 0.05). The experimental design in this study was a split-plot design with the main effect being storage temperature and the sub-effect being treatments. Each replication was represented by a single fruit. EDAX analysis revealed that Si passed through the exocarp into the mesocarp tissue in fruit treated with high concentrations of silicon, i.e., KSil 2940 ppm. Significant differences (P < 0.001) were observed in temperature means with regards to firmness. Fruit treated with KSil and NTS only and stored at 5°C were firmer than fruit stored at other temperatures. Fruits treated with Si in the form of KSil 2940 produced the least amount of CO2, while non-treated fruits (Air) had the highest respiration rate. Fruit stored at room temperature (25°C) produced significantly higher amounts of CO2 and peaked much earlier than fruit stored at other temperatures. Ethylene results showed that there were differences (P < 0.05) between temperature means with the highest net ethylene being produced by fruit stored at 25°C. There were also significant differences amongst treatment means (P < 0.001), with fruits treated with KSil 2940 ppm producing the least ethylene. There were significant differences (P < 0.001) in temperature means with regards to the total phenolic concentration with fruits stored at 1°C having the highest TP concentration (26.4 mg L-1 gallic acid). Fruit treated with KSil 2940 ppm had the highest total phenolic concentration whilst the control fruit (Air and Water) had the lowest. There were also differences (P < 0.05) in storage temperature means with respect to the total antioxidant capacity. Fruit stored at -0.5°C had the highest TAOC (52.53 μmol FeSO4.7H2O g-1 DW). There were no significant differences in TAOC (P > 0.05) with regards to treatment means although fruit treated with KSil 2940 ppm and stored at -0.5°C showed the highest TAOC of 57.58 μmol FeSO4.7H2O g-1 DW. With regards to the concentration of major sugars in avocado, mannoheptulose and perseitol (mg g-1), no significant differences (P > 0.05) were observed in temperature means. However, fruit stored at -0.5°C had the highest concentration of these C7 sugars compared with fruit stored at other temperatures. There were significant differences in treatment means (P < 0.001) showing that fruit treated with KSil 2940 ppm had the highest concentration of both mannoheptulose (18.92 mg g-1) and perseitol (15.93 mg g-1) in the mesocarp tissue. Biochemical analyses showed differences (P < 0.05) in storage temperature means with respect to PAL enzymatic activity. Fruit stored at 5°C had the highest PAL activity (18.61 mmol cinnamic acid g-1 DW h-1) in the mesocarp tissue compared with fruit stored at other temperatures. There were significant differences in treatment means (P < 0.001) with regard to PAL activity. Fruit treated with KSil 2940 ppm had the highest PAL activity (23.34 mmol cinnamic acid g-1 DW h-1). This research has demonstrated the beneficial effects, particularly applications of 2940 ppm Si in the form of KSil. This treatment successfully suppressed the respiration rate of avocado fruit. Biochemical analyses of total antioxidants, total phenolics and PAL activity in the mesocarp tissue have shown the usefulness of Si in improving the fruit’s metabolic processes. The C7 sugars (D-mannoheptulose and perseitol) also seem to be more prevalent in avocado fruit treated with Si (particularly KSil 2940 ppm) than in non-treated fruit. This suggests that an application of Si to avocado fruit can aid in the retention of vital antioxidants (C7 sugars).Item Effect of methyl jasmonate and salicyclic acid on chilling injury of 'eureka' lemons.(2010) Siboza, Xolani Irvin.; Bower, John Patrick.; Bertling, Isa.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.Item Studies of the micronutrients zinc, manganese and silicon in cucumbers (Cucumis sativus)(2010) Dominy, Andrew Peter.; Bertling, Isa.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.Item Horticultural manipulation techniques to improve yield, fruit size and quality in 'Wai Chee' litchi (Litchi chinensis Sonn.)(2010) Froneman, Izak J.; Bower, John Patrick.; Bertling, Isa.; Bijzet, Zelda.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.Item The potential of post-harvest potassium silicate dips to mitigate chilling injury on citrus fruit.(2012) Mditshwa, Asanda.; Bertling, Isa.; Bower, John Patrick.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.Item Management of avocado postharvest physiology.(2011) Blakey, Robert John.; Bower, John Patrick.; Bertling, Isa.Avocados are an important horticultural crop in South Africa, especially in the provinces of KwaZulu-Natal, Mpumalanga and Limpopo. The distance to traditional export markets, phytosanitary restrictions to lucrative markets such as China, the USA and Japan and increased competition in the European market have challenged the South African avocado industry. The industry has responded with improved logistics and shipping, a co-ordinated market access program and a global system to co-ordinate exports of avocados to the European market. To remain competitive on the global market, further improvements and innovations are required to improve the efficiency of postharvest operations. These improvements and innovations should be guided by a greater understanding of postharvest physiology. Avocados are a relatively new export crop, so there is still much to be learnt about avocado postharvest physiology and the optimisation of postharvest management. In this regard, reduced temperature storage (1°C) and modified humidity packaging (MHP) were investigated for their effect on fruit physiology and quality, the effect of a water- and ABA-infusion on ripening was examined and the effect of a cold chain break on fruit physiology and quality determined; near-infrared spectroscopy was also examined for its potential for its use in the avocado industry. As an initial study, the relationships between individual sugars, protein and oil were studied to understand the changes in avocado fruit during ripening. It was found that mannoheptulose and perseitol were the predominant sugars at harvest, but declined to very low levels during the first 10 days postharvest. The concentrations of glucose and fructose increased, while sucrose declined slightly during ripening. The concentration of protein increased sigmoidally during ripening, reflecting the increase in the ripening enzymes, particularly cellulase and polygalacturonase. The oil content fluctuated slightly during ripening. It is suggested that mannoheptulose and perseitol are important carbon and energy sources during ripening. Glucose concentration was also found to increase earlier in fast ripening fruit compared to slow ripening fruit, which is related to increased cellulase activity and may be related to the ABA functioning. Thereafter, storage and ripening trials in two consecutive seasons showed that 1°C storage and the use of MHP for 28 days reduced mass loss, water loss, ethylene production, respiration, softening and heptose consumption, without appreciably affecting fungal rots, physiological disorders or external chilling injury, compared to fruit stored at 5.5°C and regular atmosphere respectively. Also, the storage of fruit in MHP delayed the rise in the activity of cellulase during ripening, compared to fruit not stored in MHP, but there was no significant difference in the peak activity of cellulase, polygalacturonase or pectin methylesterase. In a separate experiment, fruit ripening was significantly affected by the infusion of ABA in an aqueous solution. Water slightly reduced the variation in ripening while ABA reduced the time to ripening and the variation; it is suggested that water stress and ABA are intrinsically involved in the ripening processes and may act as a ripening trigger. The water concentration in fruit was measured non-destructively using reflectance NIR; this model was used to determine the maturity of fruit and the loss of water during cold storage. In the cold chain break experiment, it was found that although fruit recovered after a cold chain break, in terms of ethylene production and respiration, there was a loss in quality because of severe shrivelling as a result of increased water loss. Fruit that were stored at 1°C were generally of a better quality at ripeness, if the cold chain was broken, compared to fruit stored at 5.5°C. In a follow-up experiment, it was found that significant changes occurred in avocado physiology over a 6h period. The respiration rate of fruit significantly increased after 4h at room temperature and mannoheptulose declined by 32% in control fruit and by 16% in ethephon-treated fruit after 6h. This demonstrates the potential for quality loss in a short amount of time. Furthermore, a model of avocado ripening is proposed, outlining the role of water, ABA, ethylene, respiration, ripening enzymes and individual sugars. This study has contributed to the understanding of avocado postharvest physiology and should aid in better management of avocados for improved fruit quality and consumer satisfaction.Item Enhancement of 'Hass' avocado shelf life using ultra-low temperature shipping or 1-MCP treatment and cold chain management.(2011) Kok, Richard Dean.; Bower, John Patrick.; Bertling, Isa.Avocados are becoming an increasingly important crop in South Africa, where the main producing areas include Limpopo, Mpumalanga and KwaZulu-Natal provinces. The South African avocado industry faces considerable challenges including increasing competition exporting avocados, particularly to the European market. The processes involved to export avocados has markedly improved over the past two decades, however there is always room for improvement and it is necessary to remain competitive on a global scale. Issues such as fruit being partially soft on arrival, quality defects and cold chain management breakdown are still present. It is necessary to investigate new aspects of cold storage such as extending the storage period and understanding the physiological aspects involved. To improvement such issues, an investigation was conducted on ultra-low temperature shipping (1°C) as well as the use of 1-MCP; the implementation of deliberate cold chain breaks to achieve a better understanding as to the quality influences involved; an extended storage period of 56 days to assess the quality issues and benefits involved; as well as investigating the physiological aspects involved with all above treatments on 'Hass' avocados. An initial study saw early-, mid- and late-season 'Hass' avocados stored at 1°C or 5.5°C for 28 days. Additional treatments included fruit treated and not treated with 1-MCP as well as waxed and unwaxed fruit. Storage at 1°C was comparable with 1-MCP treatment for both fruit softening in storage and extending the ripening period. Storage at 5.5°C resulted in partial in-transit ripening, if 1-MCP was not used. Early-season fruit incurred the most external chilling injury but overall levels were minimal and not concerning. Mid-season fruit were the most sound in terms of quality. It is suggested that 1°C can be used as a viable economic alternative to 1-MCP for long distance shipping of 'Hass' up to 28 days. The cold chain break trial included a 24 hour delay before cold storage, a deliberate 8 hour break at day 14 of cold storage where fruit were removed from cold storage and a control of 28 days cold storage where no break was involved. Early-, mid- and late-season 'Hass' avocados were stored at 1°C or 5.5°C for 28 days. Additional treatments included fruit treated and not treated with 1-MCP as well as waxed and unwaxed fruit. It was found that cold chain breaks do influence the amount of water loss, fruit softening and days taken to ripen. Storage at 1°C did not entirely negate the effects of cold chain breaks compared with 5.5°C, but did result in fruit which were harder at the end of storage and took longer to ripen. The use of 1-MCP also had advantageous effects with respect to significantly lengthening the ripening period, even when a cold chain break occurred, compared with fruit not treated with 1-MCP. As results of the study differed in some respects to those of previous studies, it is recommended that further work be conducted to determine what fruit or pre-harvest factors affect the fruit physiological changes which take place when cold chain breaks occur. Having the option to make use of an extended storage period would be of benefit to the industry if delays occur and fruit have to be maintained under cold storage. Extended storage of South African avocados, especially at the end of the season would also allow for the option of strategically holding back fruit from the export market in order to extend the supply period. It would not only benefit export options, but would also be highly beneficial to local pre-packers, as it would reduce the need to import fruit from the Northern hemisphere production areas during the South African off-season. Early-, mid- and late-season 'Hass' avocados were stored at 1°C or 5.5°C for 56 days. Additional treatments included fruit treated and not treated with 1-MCP as well as waxed and unwaxed fruit. The combination of 1°C with the use of 1-MCP resulted in a good shelf life as well as maintenance of internal quality and integrity. External chilling injury is of concern for early-season fruit, however, mid- and late-season fruit did not incur extensive damage. It is, therefore, advised that fruit placed in extended storage are marketed through the 'Ready ripe' program to mask any chilling injury on the 'Hass' fruit. Avocados are renown as a "healthy food" due to their nutritional value as well as containing relatively high concentrations of antioxidants. The fruit also contain high amounts of C7 sugars which can act as antioxidants. Additionally, C7 sugars and other antioxidants play important roles in fruit quality. Therefore, it is important to understand how varying storage conditions and treatments affect the levels of these physiological parameters. Treatments of cold chain break/delay included a deliberate 8 hour break at day 14 of cold storage where fruit were removed from cold storage, a 24 hour delay before cold storage and a control of 28 days where no break was involved. A 56 day extended storage period was also used. Early-, mid- and late-season 'Hass' avocados were stored at 1°C or 5.5°C for 28 days. Additional treatments included fruit treated and not treated with 1-MCP as well as waxed and unwaxed fruit. The use of 1-MCP maintained higher levels of antioxidants, ascorbic acid and C7 sugars for both the 28 day and the 56 day storage periods. The 24 hour delay had a tendency to increase consumption of anti-oxidant and sugar reserves. The use of 1°C resulted in antioxidant and ascorbic acid levels decreasing while maintaining higher sugar levels. Overall, high stress imposed on fruit decreased reserves resulting in poor quality fruit. The use of 1°C and 1-MCP treatments maintained fruit quality.Item Influence of stockplant management on yield and subsequent rooting of cuttings of cold-tolerant Eucalyptus grandis x E. nitens clones.(2012) Ziganira, Matabaro.; Bertling, Isa.; Blakeway, Felicity Clare.; Mokotedi, Mompe Edward Oscar.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.Item Some ecophysiological aspects of cashew (Anacardium occidentale L) with emphasis on possible flower manipulation in Maputaland.(1994) Roe, Denis John.; Wolstenholme, B. Nigel.There has been interest in developing a cashew industry in Maputaland, the far north-eastern corner of Natal/KwaZulu. Flowering and fruit development coincide with a rainy period, with accompanying serious flower diseases (Oidium anacardii and Colletotrichum gloeosporioides). Glasshouse studies were carried out at Pietermaritzburg, concurrently with field trials in Maputaland, in an attempt to manipulate flowering and growth of cashew trees. Two glasshouse trial were carried out. A factorial design with treatments 0, 3, 6 and 9 weeks of low temperatures (24°C day/9°C nights)(factor A) and 0, 3, 6 and 9 weeks of water deficit (Factor B) was used, with both factors in all combinations. During the second season the durations were increased to 0, 4, 8 and 12 weeks for both factors. No flowering occurred in this trial. Tree growth was not affected significantly by drought and/or cold duration. Temperature appeared to be the dominant factor at low temperatures, stomatal conductance and transpiration being suppressed by cold regardless of soil water potential. At more optimum temperatures for growth, stomatal conductance was dependent on soil water potential (r² = 0.756). Starch levels in the roots, dry matter production in the leaves, roots and stems, as well as leaf area were decreased significantly (P≤0.01) with increasing low temperature duration. Another glasshouse trial to test the effects of foliar urea at concentrations of 0, 1, 2, 4 and 8 g urea 100 l⁻¹, applied once, twice or thrice at fortnightly intervals was undertaken. The treatments were applied in late autumn/early winter of 1990 and 1991. Tree growth and flowering were monitored, and starch and leaf NH₃/NH₄⁺ analyses carried out. The highest urea concentration (8%) resulted in leaf scorch and abscission, extremely low stem diameter growth rates, and was too high for glasshouse trees. The starch contents of the 8% urea treatment were depleted significantly (P≤0.01) more than the other concentrations. The other urea treatments resulted in vigorous growth and high dry matter production. There were no significant effects of the number of sprays on cashew growth. Only seven trees flowered, and therefore no definite conclusions could be drawn regarding urea effects on flowering. Most hermaphrodite flowers (max. 76.8% hermaphrodite) opened soon after first anthesis of a panicle, and all terminal flowers of panicle branches were hermaphrodite. Flowers generally opened basipetally in a panicle, starting with hermaphrodite flowers and with progressively more male flowers. Urea sprays resulted in NH₃/NH₄⁺ build-up in the leaves, concentrations in flowering trees ranging from 100 to 700 μg g⁻¹ DM for approximately a month. A field trial at was carried out at Makatini Research Station to determine the effects of timing of a two month winter drought period on flowering and growth. An observational trial to determine the effects of girdling on growth and flowering was incorporated in the border rows of the irrigation trial. The trial tested five treatments (no irrigation during May and June, June and July, July and August, August and September, and a control treatment which received irrigation throughout Winter). Mean monthly temperatures were below 20°C, and mean minimum temperatures below 15°C for the 5 winter months during treatment application. There were no significant differences in tree growth, flowering, flushing, or yields between drought stressed treatments and control, indicating that, under the conditions at Makatini, autumn and winter temperature was the overriding factor controlling initial flower induction. Flowering occurred from early October (when mean temperatures exceeded 23 to 24°C) to late April (7 months - a prolonged flowering period), when mean monthly temperatures dropped below 23 to 24°C. Girdling of cashew trees in March and May, using girdle widths of 1, 5 and 10 mm was not successful in improving flowering and yields under the conditions of the trial. A field trial was carried out at Mosi Estate in Maputaland to test the following chemicals as tree and/or flower manipulators: foliar applied ethephon (50,100,200,500,2000 mg l⁻¹), KNO₃ (1 %, 2%, 4%), urea (1 %, 2%, 4%) and paciobutrazol (500, 1000, 2000 mg l⁻¹). A phenological model for cashew in Maputaland showed a dormant period during winter, followed by a generative flush, from which panicles and flowers were produced (peak November-January). The harvest period peaked in February and March. A strong post-harvest flush preceded the winter dormant period. Trunk starch levels were at their highest after the dormant winter period, and at their lowest following the harvest. Ethephon at high concentrations (500 and 2000 mg l⁻¹) resulted in excessive leaf drop, disturbed the root shoot balance and normal phenological patterns, and gave poor yields. The best ethephon concentrations were 100 to 200 mg l⁻¹. KNO₃had no significant effect on tree growth, flushing, flowering or yields when compared to control trees. Urea at 2% concentration gave a significant increase (P≤0.05) in flushing and simultaneous decrease in flowering. Paclobutrazol at 500 to 2000 mg l⁻¹ resulted in significantly lower growth rates, and early panicle production. The mean yields of all paclobutrazol treated treatments were higher than controls, despite a hail storm which damaged the flowers. From results of this trial, the use of these chemicals to improve yields and manipulate flowering may not be economically justified. The most promising chemical for further research was paclobutrazol.Item Effect of systemic resistance inducers applied pre- and postharvest for the development of a potential control of colletotrichum Gloeosporioides on Persea Americana (Mill.) CV 'Fuerte'.(2012) Bosse, Ronelle Joy.; Bertling, Isa.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.Item The cascade of physiological events leading to chilling injury : the effect of post-harvest hot water and molybdenum applications to lemon (citrus limon) fruit.(2012) Mathaba, Nhlanhla.; Bertling, Isa.; Bower, John Patrick.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.Item Ultra-low temperature shipping and cold chain management of 'fuerte' avocados (Persea americana Mill.) grown in the KwaZulu-Natal Midlands.(2011) Lutge, Andre.; Bertling, Isa.; Bower, John Patrick.‘Fuerte’ makes up 25% of the avocados exported from South Africa to European markets and requires shipping periods of up to 28 days and a correctly managed cold chain. A temperature of 5.5°C and expensive CA and 1-MCP treatments are currently used to delay ripening over this lengthy cold chain; however, fruit still appear on the European market showing signs of softening and physiological disorders. Increased competition on the global market and the disadvantage of a particularly long distance to the European market has challenged the South African export industry. These challenges have necessitated improved road and sea transport logistics, co-ordination with producing countries which supply fruit to European markets over similar periods as South Africa, and research into ultra-low temperature storage to possibly enable future access to new lucrative markets in the USA, China and Japan. It is also known that there are various ‘weak links’ in this cold chain and that cold chain breaks are detrimental to fruit quality, but further research into the negative effects of these cold chain breaks at ultra-low temperatures was needed. Thus, the objective of the study was to determine the potential for shipping ‘Fuerte’ avocados at temperatures of 2°C as well as determining the effects of cold chain breaks on fruit quality, throughout the growing season and possibly for an extended period of 56 days. ‘Fuerte’ avocados were harvested at three different maturity stages reflecting early-, mid- and late-season fruit, with moisture contents of 74%, 68% and 63%, respectively. Fruit were stored at 2°C or 5.5°C, treated with 1-MCP and waxed. Additionally cold chain breaks (24 hour delay and break at 14 days) were implemented. Fruit softening, mass loss, days-to-ripening, external and internal quality as well as antioxidant levels and total sugar levels were determined. The first aim was to determine whether a lower than currently used storage temperature could be a successful alternative to 1-MCP use. A storage temperature of 2°C provided good internal quality as well as reduced mass loss and fruit softening, which is related to the slightly reduced use of C7 sugars at 2°C compared with 5.5°C. Although the overall occurrence of external chilling injury was relatively low, 2°C storage caused a notably higher occurrence of external chilling injury than 5.5°C storage, particularly early in the season, but extended the days-to-ripening. Unfortunately, no correlation between the anti-oxidants in the exocarp and external damage was found. Waxing significantly reduced the external damage on fruit stored at 2°C, so much so, that the treatment combinations of ‘2°C, no 1-MCP, waxed’ showed no external chilling injury throughout the season. Further, waxing fruit at 2°C could eliminate the need for 1-MCP, delivering a product of the required shelf-life and quality. Best results were achieved for mid-season fruit stored at 2°C. Late-season fruit would potentially be the most profitable to store at this low temperature, however, body rots (anthracnose and stem-end rot) were more common in the late-season. Storage at 2°C can therefore maintain the internal quality over a storage period of 28 days and be a potential alternative to 1-MCP use as the season progresses. The effect of cold chain breaks on fruit quality was then investigated and showed that both a delay and a break in the cold chain increased mass loss and fruit softening, reduced days-to-ripening and increased external chilling injury, especially early in the season. Water loss was the main contributor to the decreased fruit quality which resulted from the delay in cooling, increasing external damage significantly, particularly early in the season. The break at 14 days had a marked effect on physiological activity of fruit during storage, seen mainly in the increased metabolic activity, resulting in increased fruit softening and water loss during storage and a decrease in C7 sugars and thus shelf-life, particularly for fruit stored at 5.5°C. Importantly, 1-MCP use and storage at 2°C reduced the effects of cold chain breaks with respect to fruit softening, however, lowering the storage temperature had a greater negating effect than 1-MCP and could be a successful alternative to the use of 1-MCP. The internal quality throughout the experiment was very good, with few internal disorders and no significant treatment effects on internal quality and C7 sugar concentrations. Overall, a break in the cold chain, before and during cold storage, resulted in a marked reduction in fruit quality. The storage temperature of 5.5°C should not be used for a 56 day storage period as it resulted in significant fruit softening during storage, even when 1-MCP was used, and resulted in significantly more external chilling injury in the mid- and late-season than at 2°C. Storage of 1-MCP treated, waxed fruit at 2°C, resulted in the best shelf-life and fruit quality, particularly mid-season fruit which had negligible external chilling injury and 100% sound fruit. Early-season fruit suffered significant external chilling injury at 2°C and late-season fruit had the highest body-rots and internal disorders at this storage temperature. Although mid-season fruit could be successfully stored at 2°C for 56 days, the use of a 56 day storage period is not recommended as a practical storage period, due to the high risk of external damage, particularly if maturity levels are not optimum and trees and fruit are not of the highest quality. Overall this thesis has shown that 1-MCP treatment can play an important role early in the season when fruit are susceptible to external damage, however, storage at 2°C results in good quality fruit and, when used in conjunction with waxing, appears to be a viable alternative to the use of 1-MCP, particularly later in the season. Further, the negative effects of cold chain breaks on fruit quality have been demonstrated and, importantly, the storage temperature of 2°C negates the fruit softening effects of these breaks, even if 1-MCP is not used.Item Yield and quality parameters of tomato cultivars as affected by different soilless production systems and beneficial micro-organisms.(2013) Maboko, Martin Makgose.; Bertling, Isa.; Du Plooy, Christian Phillipus.Most tomato cultivars used for commercial food production are imported into South Africa. Optimal growing conditions for these specific cultivars need to be determined, as wrong cultivar choices can lead to great financial losses. Lack of information on selecting well-performing cultivars may lead to lower yield or unacceptable fruit quality. Information on the performance of tomato cultivars under South African conditions, utilizing plastic tunnels or shadenet structures under soilless cultivation is still very limited. Soilless cultivation of vegetables is becoming a preferable over in-soil cultivation due to the improved yield and quality of produce, efficient water and nutrients usage by the crop; furthermore, the grower can regulate nutrient solution, electrical conductivity and pH of the nutrient solution. To identify the optimal system for growing tomatoes hydroponically, the performance of four tomato cultivars (‘FA593’, ‘Miramar’, ‘FiveOFive’ and ‘Malory’) under different growing conditions was evaluated: directly planted in soil under 40% shadenet with drip irrigation, a closed hydroponic system under 40% shadenet, an open bag system under 40% shadenet, or an open-bag system in a temperature controlled as well as a non-temperature controlled tunnel. The study revealed that ‘Miramar’ performed better than the other cultivars in all production systems, with the exception of soil cultivation where there were no differences amongst the four cultivars. Fruit cracking was found to be directly correlated with fruit size, as the large-sized cultivars ‘Malory’ and ‘FA593’ were more susceptible than the other two cultivars. Plants grown under shadenet were prone to fruit cracking and raincheck as well as early blight. Higher yields were obtained when plants were produced in the open bag system under temperature controlled conditions and in the closed system under shadenet. Growing tomatoes in the non-temperature controlled tunnel resulted in high incidences of fruit cracking, poor yield and pre-mature fruit ripening probably due to high and fluctuating temperatures under such conditions. The average marketable yield was 88% and 59% of the total yield in the temperature controlled and non-temperature controlled tunnels, respectively. A further experiment was carried out to improve yield and quality of tunnel tomatoes using beneficial micro-organisms, i.e., arbuscular mycorrhiza fungi (AMF) at different nutrient concentrations. Tomato seedlings were treated with Mycoroot™ containing four mycorrhiza species (Glomus etunicatum, Paraglomus occultum, Glomus clarum and Glomus mossea) at transplanting and subsequently transferred to either a temperature controlled or a non-temperature controlled tunnel under the recommended (100%) or reduced (75 and 50%) nutrient concentrations. Sawdust was used as a growing medium in this experiment. Application of AMF neither enhanced plant growth, yield, nor fruit mineral nutrient concentrations; although fruit Mn and Zn concentrations in the temperature controlled tunnel increased significantly following AMF application. Plants grown in the non-temperature controlled tunnel had significantly poorer plant growth, and lower yield and lower fruit mineral concentrations, compared with fruit from plants in the temperature controlled tunnel. Tomato plants in the non-temperature controlled tunnel had higher levels of micro-elements in leaf tissue, compared with those in the temperature controlled tunnel. The highest yields were obtained from plants fertigated with 75% of the recommended nutrient concentration, as compared with the 100 and 50% nutrient concentrations. When coir was subsequently used as the growing medium, Mycoroot™ applied at seeding and transplanting did not enhance mycorrhizal colonization or fruit quality. Growing tomatoes under reduced nutrient supply reduced the total soluble solids in the juice of the fruit, but improved total and marketable yield, as well as the number of marketable fruit. This effect was more substantial in the temperature controlled than in the non-temperature controlled tunnel. Fruit firmness and leaf chlorophyll concentrations were significantly higher in plants grown in the temperature controlled tunnel. Growing tomatoes in sawdust improved the leaf Mn and Ca concentration over that of tomato plants grown in coir. Mycorrhizal colonisation did not have a beneficial effect on tomato yield and quality. The study indicated that cultivar selection was important in obtaining the highest yield and quality of tomato using the closed hydroponic system under shadenet and the open bag hydroponic system in the temperature controlled tunnel. Temperature controlled tunnels with a pad–and-fan cooling system are still an effective way of cooling the tunnel environment which resulted in high yield and high quality of tomatoes with a higher fruit mineral content than that obtained under non-temperature controlled conditions where only natural ventilation is relied on. Results also demonstrated that mycorrhizal colonization in soilless condition has limited beneficial effects in allowing for better nutrient uptake and thereby for improved yield and quality of tomatoes. Further studies, including different media, nutrient composition and concentrations, need to be carried out to investigate the possible causes of AMF failure to improve yield, despite good AMF root colonization.Item Methyl jasmonate and salicylic acid enhance chilling tolerance in lemon (citrus limon) fruit.(2013) Siboza, Xolani Irvin.; Bertling, Isa.; Odindo, Alfred Oduor.South African ‘Eureka’ lemon fruit must be exposed to chilling temperatures (± 0.6°C) as a mandatory quarantine treatment against insect pests for all its overseas markets. Chilling lemon fruit at such temperatures may develop chilling injury (CI) symptoms on the flavedo. This negative effect on fruit quality reduces fruit marketability. This study evaluated postharvest factors influencing physiological, biochemical and ultra-structural mechanisms involved in alleviating CI in lemon fruit. It was hypothesised that treatment with methyl jasmonate (MJ) and salicylic acid (SA) may enhance chilling tolerance in lemon fruit by maintaining cellular integrity and inducing synthesis of enzymatic and non-enzymatic antioxidants. Furthermore, fruit susceptibility to CI was associated with the source of fruit. Lemon fruit were harvested from three locations representative of moderate subtropical, warm temperate and cool subtropical environments. Harvested fruit were treated either with 10 μM MJ, 2 mM SA or 10 μM MJ plus 2 mM SA, stored either at -0.5, 2 or 4.5°C for 0, 7, 14, 21, or 28 days and afterwards transferred to 23°C for a week as shelf-life simulation. Thereafter, fruit were evaluated for alterations in physiological, biochemical and ultra-structural features involved in the manifestation of CI symptoms. Chilling damage was more severe in untreated lemon fruit than in treated lemon fruit. Storing lemon fruit at 4.5°C accelerated the manifestation of CI symptoms more so than at 2°C while storage at -0.5°C delayed the manifestation of CI symptoms. Lemon fruit of moderate subtropical origin were more chilling-tolerant than lemon fruit of warm temperate and cool subtropical origin. Treatment with 10 μM MJ plus 2 mM SA significantly (P < 0.05) improved chilling tolerance in lemon fruit. This treatment effectively maintained membrane integrity, thereby retarding electrolyte leakage and membrane lipid peroxidation as well as mass loss and respiration rate. Treatment with 10 μM MJ plus 2 mM SA was also effective in enhancing the antioxidant concentrations of vitamin E and carotenoids. The production of these antioxidants could have been part of a defence system against chilling damage, reducing CI and maintaining fruit quality. Treatment with 10 μM MJ plus 2 mM SA enhanced the concentration of compounds involved in chilling resistance, such as proline, soluble sugars, ascorbic acid and total phenolics as well as the enzyme phenylalanine ammonia-lyase (PAL). The enhancement of the defence mechanisms may have played a role in enhancing chilling tolerance in lemon fruit. The treatment also inhibited certain enzymes involved in tissue browning, such as peroxidase (POD) which might have contributed to delaying manifestation of symptoms. Polyphenol oxidase (PPO) was found to not be a good biochemical marker of the occurrence of CI. Treatment with 10 μM MJ plus 2 mM SA appeared to be able to enhance chilling tolerance in lemon fruit by maintaining the ultra-structure of the cuticle, cell wall integrity, cell membrane of parenchyma cells of the flavedo. This treatment also preserved the mineral nutrients of the flavedo (carbon, oxygen, phosphorus, potassium, calcium, magnesium, sulphur, sodium, silicon and aluminium) during cold storage. This could have played a role in protecting the fruit against chilling stress and maintaining fruit quality. Treatment with 10 μM MJ plus 2 mM SA reduced ROS production, while the activity of enzymatic antioxidants such as catalyse (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR), and accumulation of essential proteins was enhanced. This increase in activity of enzymatic antioxidants and the presence of stress-responsive proteins in the lemon flavedo could have been directly involved in enhancing chilling tolerance. The CI symptoms were accompanied by an increase in membrane permeability, membrane lipid peroxidation as well as phospholipase D (PLD) and lipoxygenase (LOX) activity; however, treatment with 10 μM MJ plus 2 mM SA effectively reduced the membrane permeability, membrane lipid peroxidation, and PLD and LOX activity induced by the cold treatment. This could have contributed to the efficacy of 10 μM MJ plus 2 mM SA in inhibiting the manifestation of CI symptoms. Treatment with 10 μM MJ plus 2 mM SA enhanced flavedo total antioxidant capacity measured by ferric reducing ability of plasma; 2,2-diphenyl-1-picrylhydrazyl; 2,2-azinobis (3-ethyl-benzothiazoline-6-sulfonic acid) and the oxygen radical absorption capacity assays. The enhancement of antioxidant capacity in lemon flavedo could have contributed to the fruit’s chilling tolerance. Therefore, the effect of 10 μM MJ plus 2 mM SA treatment, enhancing chilling tolerance, may be attributed to its ability to enhance enzymatic and non-enzymatic antioxidants; activate essential proteins and mitigate the effect of ROS accumulation. With the use of 10 μM MJ plus 2 mM SA treatments, the South African citrus industry will be able to meet the quarantine temperature requirements for exportation of lemon fruit whilst reducing economic losses, depending on the preharvest conditions experienced by the fruit in each shipment.