Global warming is a major concern to the world’s population. It is caused by greenhouse gases that result from the burning of fossil fuel. The fossil fuel reserves are rapidly depleting as the needs and wants of man in the world increases. Biodiesel is one of the solutions proposed to remedy this environmental crisis facing the world today. The aim of this study was to characterise the biodiesel that can be produced from the oil of Litsea glutinosa by transesterification. Biodiesel can be used in a diesel engine without modification and be produced from many different natural renewable oil sources such as algae, plants and kitchen waste material. Jatropha curcas has been identified as a potential producer of oil for biodiesel. The biodiesel properties of Jatropha curcas meet the required American Society for Testing and Materials (ASTM) standards. The fruit of Jatropha curcas contains 40.0% lipids. The oil has a saponification number of 202.6 and an iodine value of 93.0. However Jatropha curcas cannot be grown in South Africa because it is a highly invasive plant. Cetane number is the most important parameter of biodiesel. The higher the cetane value, the better the quality of the biodiesel. Oil from Jatropha curcas has a cetane number of 57.1. An alternative is the oil from Litsea glutinosa, which is found as a naturalised free forest along the South African coastline, and is also found in many Asian countries. It has many medicinal properties, however, it is not edible and hence its use for biodiesel does not add to the debate of fuel versus food production. The cetane number of oil from Litsea glutinosa is 64.79, which is ideal for ignition, and the fruit with 61.29% lipids can yield valuable quantities of biodiesel. Thus, the aim of the research was to determine the potential of Litsea glutinosa as a source of biodiesel. Furthermore, to maintain a sustainable source, Litsea glutinosa was micropropagated, and transformation of Litsea glutinosa was attempted for hairy root cultures. The Clevenger apparatus was used to extract fatty acids from dried crushed fruit of Litsea glutinosa. Fatty acids were converted to fatty acid methyl esters by transesterification. Transesterification was conducted in the presence of nitrogen and the reaction was catalysed with a mixture of methanol and sodium hydroxide (NaOH). The ratio 1 : 3 of oil to catalyst mixture was used for optimum transesterification to ensure a forward reaction and it was transferred to a separating funnel to allow the glycerol and fatty acid methyl esters to separate. GC-MS was used to determine the fatty acids. The iodine number, saponification number, acid value, viscosity, kinematic viscosity, density, specific gravity, thermostability, distillation point and sulphur content were determined. The seeds of Litsea glutinosa were germinated and tissue culture callus was produced from the seeds and leaves. The leaves and stems were used to produce hairy root cultures by inoculating them with Agrobacterium rhizogenes. Litsea glutinosa yielded 61% oil, which included 47 fatty acids in the fruit and 24 fatty acids in the seeds. The fatty acid profile of the oils indicated that the predominant fatty acids present were those that are essential for good quality biodiesel. The dominant fatty acids found in the fruit were 65.4% 9-octadecenoic acid and 13.6% hexadecanoic acid. The dominating fatty acids found in the seeds contained 36.3% 9-octadecenoic acid, 13.9%, hexadecanoic acid and 39.1%, dodecanoic acid. The iodine value was 6.3. The saponification value was 274. The acid value was 0.45 mg KOH. g-1. The viscosity was 22.48 mm2. s-1 and the kinematic viscosity was 23.84 mm2. s-1. The density was 942.69 kg. m-3 and the specific gravity was found to be 0.9 g. cm-3. The distillation temperature ranged between 52.2°C to 610.2°C. The sulphur content was found to be 383 µg. ml-1. These characteristics indicate that Litsea glutinosa can be used as a source of biodiesel, because the properties meet the required ASTM standards. However, the production of biodiesel from Litsea glutinosa has not been commercialised because the production of fuel is dependent on the fruit of the plant, which is seasonal. To overcome this, a part of this study investigated micropragation of Litsea glutinosa and transformation of Litsea glutinosa by Agrobacterium rhizogenes into hairy roots and attempts where made to determine whether fatty acid could be produced by these techniques. Callus cultures were grown on MS media and McCowns woody plant media supplemented with 1 ml BAP and 1 ml 2,4-D per 1 L of media. Callus cultures were obtained in the light. However, Litsea glutinosa resisted transformation by Agrobacterium rhizogenes.
Regeneration of plants from callus cultures of roots induced by Agrobacterium rhizogenes on Alhagi pseudoalhagi