The Etlingera elatior is a tropical perennial and rhizomatous herbaceous plant that possesses beautiful flowers with different colors and a great ornamental appeal. To produce new cultivars it is necessary crossings between existing varieties which are rare for this species. Among the difficulties found to grow Etingera elatior is the acquisition of quality seedlings with appropriate sanitary standards to supply the demand of the producers. Another considerable problem is the seedlings production that, usually made by bunch division, brings high costs and disease problems. This work aimed to evaluate the viability of the pollen grains in flowers of the Red Torch, Pink Torch and Porcelain cultivars to be used in a future breeding program to create new commercial types of the Etlingera elatior, and developing a protocol for multiplication in vitro of Etlingera elatior cv. Red Torch. Therefore, it was carried out an study to check the viability of the pollen grains in open and closed flowers of small, medium and tall stick flowers. The pollen grains were stained in a lugol solution and observed on a stereoscopic microscope. The micropropagation potential was evaluated by testing the effects of 6-benzilaminopurine alone in different concentrations (1, 2 e 3 mg.L-1) the germination of zygotic embryos and the micropropagation of the explants from in vitro germinated seeds, or combined to kinetin or naphtalenacetic acid in the in vitro multiplication of the explants. The percentage of stained pollen grains above 99%, showed that no effect of the cultivars, size of the sticks flowers or aperture of the flowers was observed, indicating that pollen viability is not the cause of the low fruit formation in this species. The largest number of sprouts (7,86) was obtained in a MS medium culture enriched with 3 mg.L-1 BAP. In the absence of hormones the explants showed the highest sprouts length, roots number and length. The acclimatization of the explantes at a glasshouse presented a satisfactory rate (>80%) of plant set, and after 60 days they were ready to be transported to the field. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O bastão do imperador (Etlingera elatior (Jack) R.M. Smith) é uma planta tropical herbácea, rizomatosa e perene, que possui inflorescências belas e vistosas em diferentes tonalidades, possuindo um grande apelo ornamental. A criação de novas cultivares de bastão do imperador necessita de cruzamentos intervarietais, pela formação de frutos e sementes viáveis, os quais são raros nessa sp. Um outro problema considerável para o seu cultivo está na produção de mudas, que quando feita convencionalmente por divisão de touceiras, acarreta elevados custos e problemas fitossanitários. Este trabalho objetivou avaliar a viabilidade dos grãos de pólen de inflorescências dos cultivares Red Torch, Pink Torch e Porcelana para a utilização em futuros cruzamentos intervarietais e avaliar o potencial micropropagativo do bastão do imperador. A viabilidade de grãos de pólen de inflorescências curtas, médias, e longas e em flores abertas e fechadas foi medida, corando-se os grãos com uma solução de lugol e visualizado-os em microscópio estereoscópico. Para o desenvolvimento de um protocolo para a micropropagação dessa espécie, testaram-se os efeitos da adição de 6-benzilaminopurina isolada em diferentes concentrações (1, 2 e 3 mg.L-1) na germinação de embriões e micropropagação de explantes in vitro, ou combinada com cinetina e ácido naftaleno acético na multiplicação in vitro dos explantes. A percentagem de grãos de pólen corados ficou acima de 99%, indicando que estes, independente do tipo de flor, do comprimento da haste da inflorescência e do cultivar, apresentam alta viabilidade polínica. Plantas oriundas de sementes produziram a maior média no número de brotos (7,86/mês) em meio de cultura Murashige e Skoog (MS) enriquecido com 3 mg.L-1 BAP. Na ausência de fitormônios os explantes apresentaram maior comprimento médio das brotações, maior quantidade e comprimento das raízes. A aclimatação dos explantes em casa de vegetação apresentou taxa de pegamento superior a 80%, e aos 60 dias estavam prontas para serem transferidas para o campo.

Diarylheptanoids are a family of plant metabolites with a characteristic structure of two hydroxylated aromatic rings attached by a linear seven-carbon chain. Diarylheptanoids have mostly been isolated from plants belonging to the Zingiberaceae family. The South African medicinal plant Siphonochilus aethiopicus, more commonly known as ‘wild ginger’, also belongs to the Zingiberaceae family. One of the compounds isolated from this plant it a novel diarylheptanoid. In this study, the synthesis of this novel diarylheptanoid will be investigated. The targeted diarylheptanoid has two substituted phenyl rings attached by a seven-carbon aliphatic chain with two sterogenic centers and a carbon-carbon double bond. Osmiumcatalysed asymmetric dihydroxylation was used to generate the two stereogenic centres. The Horner Wadsworth-Emmons (HWE) reaction was investigated, in order to generate the trans-double bond on the seven carbon aliphatic chain. HWE reaction is a transselective reaction leading to the formation of only the desired isomer. The synthetic strategy used for the synthesis of the targeted diarylheptanoid is the C2-moiety + C5-moiety strategy. The C2-moiety is the phosphonate ester and the C5-moiety is the aldehyde for the HWE reaction. In this investigation we were able to successfully synthesis the required C2-moiety and the C5-moiety. Both the precursors were synthesised from commercially available starting materials, utilising functional group transformation reactions. However, modifications to the C5-moiety were made due to its instability under the HWE reaction conditions. When the C5-moiety was an aldehyde, decomposition was seen under HWE reaction conditions. Thus the C5-moiety was converted to the corresponding lactol and then subjected to the HWE reaction. Nevertheless, this reaction was not successful, thus we were not able to couple the two precursors to form the desired seven-carbon aliphatic chain. Even though the targeted diarylheptanoid was not successfully synthesised, the synthetic route developed in this investigation is not only viable to the target compound but is also versatile enough to allow the synthesis of its analogues. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2010.

This work attempts to offer an insight into the problematic of the genus Elettariopsis Baker, the last unrevised genus in the subfamily Alpinioideae (Zingiberaceae). Phylogenetic analyses are performed on ITS, matK and DCS sequence data and correlated with absolute genome size and biogeographical distribution of the samples. Elettariopsis as a genus is found to be weakly supported and strongly supported only with the addition of some species of Amomum Roxb., including the type species A. subulatum. The absolute genome size in this group is greater than in the outgroup represented by members of the Zingiberoideae subfamily. The evidence given by sequence data further suggests that Elettariopsis is divided into two well-supported groups, the "E. curtisii" group and the "E. triloba/E. unifolia" group, each of which contains several well-supported clades. In the analysis of absolute genome size it is shown that the absolute genome size in the "E. triloba/E.unifolia" group is higher than in the "E. curtisii" group. These two groups also differ slightly in their biogeographical distribution, the group G being distributed in only in Vietnam, Laos, and Thailand, while members of group H are also occurring in Singapore and Indonesia (Borneo). Keywords: Zingiberaceae, Elettariopsis, South-East Asia, ITS,...

The variability of the genome size reaches several grades even within relatively close groups of plants. The study of the genome size in the phylogenetic context provides interesting results which characterize the evolution of the individual groups of plants. In this respect, tropical plants have yet not been studied. Tropical genus Globba (ca. 100 species) belongs to an economically significant family Zingiberaceae. The diversity centre is found in Thailand but it spreads from east India and southern China up to Indonesia and the Philippines. It is a polyploid complex which exists in two cytotypes within one genus (2n = 32 a 2n = 48); it is characteristic minimally in three out of seven distinguished sections. The aim of this thesis has been a reconstruction of the group phylogeny, discovering the role of the polyploid and evaluation of the genome size evolution of the Globba genus in the phylogenetic context. To this end, modern biosystematic methods were used (flow cytometry, chromosome counting, sequencing of the nuclear and chloroplast DNA regions). Many types of software and statistical methods were used to process and interpret the data. In this group, the genome size was measured for the first time. Out of 87 individuals, the smallest size was measured with Globba nuda (2C = 1.11 pg). The...

Siphonochilus aethiopicus (Schweinf.) B.L. Burtt (Zingiberaceae), commonly known as wild ginger, is a highly sought after plant for use in traditional medicine in South Africa. Over-exploitation of this medicinal plant has resulted in regional extinction in the wild. As a result, there is great interest in the medicinal properties of S. aethiopicus, and as a plant for small scale cultivation to increase the supply for use in traditional medicine. Water, ethanol and ethyl acetate extracts were prepared from the leaves, rhizomes and roots of S. aethiopicus. These extracts were tested for in vitro anti-inflammatory activity in the cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) assays, and in the microdilution antibacterial assay. The aqueous extracts showed no significant prostaglandin synthesis inhibition in the COX-1 and COX-2 assays. The ethanol and ethyl acetate extracts of the leaves showed the highest levels of activity at a concentration of 250 µg ml¯¹ per test solution, in both the COX-1 and COX-2 assays. The ethanol and ethyl acetate extracts of the rhizomes and roots also had moderate levels of activity in the COX-1 assay. These results provide some evidence for the rational use of S. aethiopicus in traditional medicine for anti-inflammatory purposes. In the microdilution antibacterial assay, no inhibitory activity against the test bacteria was detected with the aqueous extracts. The ethanol and ethyl acetate extracts tested showed greater antibacterial activity at minimal inhibitory concentrations ranging from 0.78 to 3.13 mg ml¯¹ against the gram-positive bacteria (Bacillus subtilis, Staphylococcus aureus) than the Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae). No distinct differences were observed between the ethanol and ethyl acetate extracts, or between the different plant parts. A serial extraction of S. aethiopicus rhizome material was conducted and the extracts were tested in the COX-1 assay and the microdilution assay as a preliminary investigation for a bulk extraction. The hexane and ethyl acetate extracts gave slightly higher COX-1 inhibition than the ethanol extract. No distinct differences were observed in the microdilution assay. A bulk ethyl acetate extract of S. aethiopicus rhizome material was prepared, yielding 6.3 g of a thin orange oil. Vacuum liquid chromatography (VLC) was used to fractionate ≈4 g of the extract. The VLC fractions were evaluated using thin layer chromatography (TLC) and a bioautographic assay, using S. aureus as a test organism. The fractions were also tested in the COX-1 assay. The bioautography revealed a number of compounds which exhibited antibacterial activity. Fraction C was purified further using preparative TLC, and 24.9 mg of a pure compound from R,0.54 (toluene:ethyl acetate 93:7) was isolated. The structure of the compound was elucidated from nuclear magnetic resonance (NMR) spectra, and mass spectroscopy of the compound was also recorded. The compound was identified as the sesquiterpenoid furanoeremophil-2-en-1-one, which is structurally identical to the recently reported compound 4aαH-3,5α,8aβ-trimethyl-4,4a,9-tetrahydro-naphtho[2,3-b]-furan-8-one. The compound showed only a very minimal bacteriostatic effect in the microdilution assay. S. aethiopicus plants were harvested before and after seasonal senescence. Ethanol extracts were prepared from fresh or dried material of the leaves, rhizomes and roots, and tested in the COX-1 assay and the microdilution assay TLC fingerprints of the various extracts were also prepared. No noteworthy changes in COX-1 inhibition, due to senescence, were observed with extracts prepared from fresh material, although there did appear to be a slight decrease in activity in the α-roots and an increase in the β-roots after senescence (fresh and dry). A decrease in the antibacterial activity of the leaves and an increase in the antibacterial activity of the α-roots was observed after senescence. These results suggest that the time of harvest may only have a minimal influence on the degree of anti-inflammatory and antibacterial activity. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2002.

Title page, table of contents and abstract only. The complete thesis in print form is available from the University of Adelaide Library. / The aim of the work described in this thesis was initially to screen the ethanol extracts of eleven Indonesian ginger species (Zingiberaceae family) for anticancer activity. MCF-7 breast and HT-29 colon cancer cells were used for the investigations. Extracts of Zingiber aromaticum and Boesenbergia pandurata were found to be the most active species, similar to that of Curcuma langa which has been shown to possess anticancer activity in vitro and in vivo (Aruna and Sivaramakrishnan, 1992; Azuine and Bhide, 1992). These two active species were then further investigated. Bioactive compounds from the species were isolated and identified using various chromatography procedures and nuclear magnetic resonance (NMR) and their anticancer activities were further tested on MCF-7 breast and HT-29 colon cancer cells including cell cycle analysis and measurements of apoptosis. The ethanol extracts of these two active species were also investigated using the AOM-induced colon cancer model in rats. The antiinflammatory activity of the ethanol extract of Z. aromaticum was also investigated using dextran sulfate sodium (DSS) induced ulcerative colitis (UC) in rats. The inhibitory activity of ethanol extracts of rhizomes of 11 ginger species was initially tested against MCF-7 breast and HT-29 colon cancer cells using colorimetric tetrazolium salt (MTT) assay. Ethanol extracts of eight species (Amommum cardamomum, C. longa, C. mangga, C. xanthorrhiza, Boesenbergia pandurata, Zingiber aromaticum, Z. officinale, Z. cassumunar) showed a strong inhibitory effect on the growth of the cancer cells with the IC50 concentrations between 100-100 g/ml. The ethanol extract of Curcuma aeruginosa was less active (IC5o between 100-120 g/ml) and extracts of Kaempferia galangal and K. rotunda had no effect on the growth of either cell lines at concentrations up to 250 g/ml. Ethanol extract of C. longa was used as a comparison since curcumin, an active compound isolated from this species, has had demonstrated its anticancer activity in vitro, in vivo and is currently undergoing clinical trial against colon cancer (Greenwald, et al., 2001; Sharma et al., 2001). Extracts of Z. aromaticum and B. pandurata had very strong inhibitory activity similar to the extract of C. longa. Curcumin was not detectable in either Z. aromaticum or B. pandurata. The ethanol extracts of the active species were not toxic on human skin fibroblast cells (SF 3169). The ethanol extracts of Z. aromaticum and B. pandurata were further fractionated using two different solvents by reversed phase preparative HPLC. Fraction A was eluted with a mobile phase containing 5% vlv aqueous methanol containing 0.025% v/v trifluoroacetic acid (TFA) and fraction B was eluted with 100% methanol. The inhibitory activity of fractions was then investigated against HT-29 colon cancer cells and assayed using the MTT assay. Zerumbone, a sesquiterpenoid compound was isolated from fraction B of the extract of Z. aromaticum and a chalcone derivative, panduratin A was isolated from fraction B of the extract of B. pandurata. Curcumin was in fraction A of extract of C. longa. The anticancer activity of zerumbone and panduratin A was investigated using MCF-7 breast. HT-29 and CaCo-2 colon cancer cells. The inhibitory activity of the active compounds was assessed using the MTT assay. The ICso of zerumbone in each of the cell lines was about 10 uM and of curcumin on HTU29 cells was 25 uM. The IC50 of panduratin A in HT-29 cells was 16 uM and in MCF-7 cells was 9 uM. Zerumbone and panduratin A showed antiproliferative effects by alteration of the DNA distribution in the cell cycle and induction of apoptosis. HT-29 cells treated with zerumbone at concentrations of 10 -25 uM or panduratin A at concentrations of 9 -65 uM for 24 h were stained with propidium iodide (PI) to determine cell cycle distribution and analysed using FACScan flow cytometry. The proportion of cells in the S phase was reduced from 18.7% in untreated cells to 10.2% in HT-29 cells after treatment with zerumbone at 10 uM to 3.1% at 25 uM. Cells in the G2 phase increased from 18.5% at 10 uM to 40% at a concentration of 25 uM. Panduratin A increased the proportion of cells in the GO/G1 phase from 33% of untreated cells to 71% after treatment with 65 uM for 24 h. Panduratin A slightly reduced the proportion of cells in S phase and cells in G2/M phase also decreased from 36,8% in untreated cells to 15.4% at 65 M. Apoptosis was determined using double labelled (Annexin-V-Fluos and PI) and then evaluated using FACScan Flow Cytometry. Morphological features of apoptosis were also examined using DiffQuick stain and fluorescent Hoechst 3355 and 4,6-diamino-2-phenylindole (DAPI). Zerumbone induced apoptosis in HT-29 cells in a dose dependent rnanner, At 48 h, 2% of cells treated with 10 M of zerumbone underwent apoptosis, which increased to 8% when treated with 50 M, Panduratin A at 28 M increased the number of cells undergoing apoptosis from 2,2% to 16.7% when treated with a concentration of 65 M. The ethanolic extracts of Z. aromaticum and B. pandurata were also investigated using the azoxymethane (AOM) induced aberrant crypt foci (ACF) model of colon cancer in rats in a short and long term study. Ethanolic extracts of C. tonga and curcumin were used as comparison. The basal diet used throughout all animal studies in this thesis was a semi-purified AIN-93 G diet (Reeves et aI., 1993). ACF were induced by two doses (15 mg/kg BW) subcutaneously of AOM one week apart and ACF were visualised in the formalin fixed colon using methylene blue stain. The ACF study was run over a short (5 weeks) and long (13 weeks) experiments. Diets containing ethanol extracts prepared from the equivalent of 2% (w/w) dried rhizome of Z. aromaticum, B. pandurate or C. tonga in a short term study did not affect the formation of ACF in rats compared to those in the control diet group. The ACF formation in a short term study was dominated by small numbers of aberrant crypts (1 or 2) per focus. It is suggested that large ACF (4 or more ACs/focus) are better predictors of colon cancer (Uchida et aI., 1997; Jenab et aI., 2001). Diets containing ethanol extracts of the equivalent of 4% by weight of dried rhizomes of Z. aromaticum, B. pandurata, C. longa were investigated over 13 week study, Total ACF were significantly reduced by Z. aromaticum extract (0.34%) in the diet (down 21%, p<0.05) relative to rats fed the control diet. A similar reduction was observed with C, longa extract (0.86%) in the diet (down 24%, p<0.01) and with 2000 ppm curcumin. There was no significant different in small ACFs (1-2 ACs/ focus) between dietary treatments. The number of foci containing 3-4 ACs/focus was significantly reduced (35%, p<0,001) in animals fed the Z. aromaticum extract and 34% (p<0.001) of animals fed the C. tonga extract. The total number of ACF containing 5 or more ACs per focus of animals fed 0.34% Z. aromaticum extract was 41 % lower than control (p<0.05) and for 0.86 % C. tonga extract was 22% (not significant). A diet containing extract (0.56%) of B. pandurata did not significantly affect the formation of ACF compared to the control AIN group. The concentration of zerumbone in the Z.aromaticum extract diet was assayed at 300 ppm, and of curcumin in the C. tonga extract diet was also 300 ppm. The concentration of panduratin A was not assayed in the diet due to late identification of the active compound. The antiinflammatory activity of ethanol extract of Z. aromaticum was investigated using dextran sulfate sodium (DSS) induced ulcerative colitis in rats. Sulfasalazine, a widely used compound to treat inflammatory bowel disease (IBD) in humans was used as the positive control. Diets containing ethanol extracts (0.34% and 0.68%) prepared from the equivalent of 4% and 8% by weight of dried rhizomes of Z. aromaticum were given to the animals throughout the experiment. On day three, rats were given 2% DSS in drinking water for 5 d and then just water for 3 d and then were killed. During the DSS treatment rats were maintained in metabolic cages, body weight, food and fluid intake and clinical symptoms such as consistency of stools and blood in faeces were recorded daily. There was slight but not significant reduction in the body weight of rats fed 0.68% extract of Z. aromaticum in the diet due to reduced food consumption. The extract of Z. aromaticum (0.34%) and sulfasalazine suppressed clinical signs of ulcerative colitis. Eleven percent of the controls were hemoccult positive on day 2 after DSS administration, which progressed further by day three with 67% being hemoccult positive and 100 % on day five. By comparison, blood appeared on day 3 of rats treated with diet containing 0.34% and 0.68% extract of Z. aromaticum and 0.05% sulfasalazine, and only 33%, 67% and 22%, of rats being hemoccult positive on day 5 respectively. The disease activity index (DAI) of rats fed diet containing 0.34% extract of Z. aromaticum was about 0.4 and similar to those which were fed with diet containing sulfasalazine. The DAI of untreated rats was 1.4. The crypt score of rats fed the extract of Z. aromaticum was slightly reduced but it was not significantly different from those of untreated rats. Other histological scores were not significantly different between dietary treatments. Extract of Z. aromaticum significantly decreased the content of PGE-2 in colon tissue compared to that of untreated animals. There was a reduction of TX8-2 content in colonic tissue of rats fed with extracts of Z. aromaticum but this was not significant. The activity of myeloperoxidase (MPO) activity in the colonic tissue of rats fed with sulfasalazine was significantly lower than that of the untreated controls and those which fed with extracts of Z. aromaticum. The results from the studies performed in this thesis showed that extract of Z. aromaticum which contains an active sesquiterpenoid zerumbone have anticancer and antiinflammatory activity suggesting that the extract may have benefits as a chernopreventative agent. However further studies are needed to elucidate their other pharmacological actions. Panduratin A showed potential anticancer activity in cell culture in vitro. However an extract of B. pandurata did not have effect on the AOM-induced colon cancer model. Different cancer models such as breast and prostate cancer could be used to further investigate the anticancer activity of extract of B. pandurata and panduratin A and to elucidate their mechanism. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1097849 / Thesis (Ph.D.) -- University of Adelaide, Dept of Medicine, 2003

The species of Amomum Roxb. (Zingiberaceae) in Cambodia, Laos and Vietnam are revised. Thirty-five species and two varieties are recognised, all names are typified, and detailed descriptions and a key are provided. Nine new species are described and one species is validated. Whilst revising Amomum for the Flore du Cambodge, du Laos et du Viêtnam, we have proposed to conserve the name Amomum villosum Lour. with a recent collection from Laos, which was not included in the protologue, as its type. Our research on the use of Amomum focuses on the use of plants during pregnancy, parturition, postpartum recovery and infant healthcare among three ethnic groups, the Brou, Saek and Kry. The investigations aim to identify culturally important traditions that may facilitate implementation of culturally appropriate healthcare. Data were collected in Khammouane province, Lao PDR, through group and individual interviews with women by female interviewers. More than 55 plant species are used in women's healthcare, of which &gt; 90 % are used in postpartum recovery. This wealth of novel insights into plant use and preparation will help to understand culturally important practices such as confinement, dietary restrictions, mother roasting and herbal steam baths and their incorporation into modern healthcare. Through chemical analyses of Amomum we have recorded compounds with antimicrobial, analgesic and sedative effects that point to an empirical development of the traditional treatments around childbirth. Essential oils of three species used in hotbed and mother roasting, Amomum villosum Lour. Amomum microcarpum C.F.Liang &amp; D.Fang and Blumea balsamifera (L.) DC. were found to contain significant amounts of the following terpenes: b-pinene, camphor, bornylacetate, borneol, linalool, D-limonene, fenchone, terpinen-4-ol and a-terpinene.

This study examines field cropping constraints for domestication of an endangered, wild medicinal plant, Siphonochilus aethiopicus, (Schweif.) B.L. Burt. Extensive literature review and careful observations of plant growth behavior during two years of crop trials overturned several long-held but erroneous claims that have consistently appeared in the scholarly literature, and revealed previously undocumented plant growth characteristics. S. aethiopicus (Schweif.) B.L. Burt. is a rhizomatous corm, not a rhizome. Field growth observations demonstrated clearly that the false stem and leaves grow continuously from emergence in September to senescence in April-May; the corm retains its tuberous roots during winter senescence, and is genetically preprogrammed to shoot in September. Flowers may emerge throughout the growing season (not only initially prior to shoot emergence), typical leaf count is 11-15, not 6-8 as previously reported, numbers that remain constant even when the plant height increases by 20-30% under shade, and leaf distichy is independent of the sun’s course and is unaffected by mother corm orientation. S. aethiopicus proved to be unusually resistant to common field diseases and pests, and resilient to severe hail. The responses of S. aethiopicus were tested in a series of field trials to the effects of levels of compost, field spacing, size of planting material, addition of biocontrol agents, different degrees of shading, and factorials of the macronutrients Nitrogen, Phosphorous and Potassium. Spacing-Composted chicken litter combinations were tested in 2005-2006 in factorial combination with Spacing at 15 cm-4.5 kg ha-1, 20 cm-7.5 kg ha-1, 30 cm-10 kg ha-1, and 40 cm-15.5 kg ha-1, and these treatments were randomized with 4 Corm planting sizes (height by base diameter in mm): Small (S, 12.38 mm x 12.6 mm), Medium Small (MS, 29.65 mm x 27.93 mm), Medium Large (ML, 38.48 mm x 37.78 mm) and Large (L, 52.37 mm x 44.10 mm). 2005-2006 ANOVA tests showed significant differences between Spacing-Compost and Corm Size for the total harvest biomass measure, with 30 cm and 40 cm spaces better than 15 cm spacing, and Corm Size MS, ML and L all better than S, and ML better than MS. Total Corms harvested per block and ii Survival Percentage were similarly significant for Corm Size, but not Spacing. Corms smaller than the Small criteria were raised separately, under optimal conditions in a nursery. In a separate 2005-2006 Compost-only trial ANOVA tests did not find significant differences between compost levels. In 2006-2007 we tested Spacing separately at 5, 10, 15, 20, 30 and 40 cm between planted corms in each plot. We tested Compost levels separately, with 0, 5, 10 and 15 kg ha-1 compost per plot. In 2006-2007 only the ML and L sizes were used in an even mix. There were no significant differences between treatments due to high experimental error, but measurement across all production parameters showed a clear trend towards best performance at spacing between 20 and 40 cm. Overall the results from the Spacing, Compost-level and Corm Size trials suggest that 30 cm is perhaps the optimal field spacing, higher compost levels tend to give better results, and the ML and L corm sizes perform better in open-sun field trials. These parameters are recommended for further field studies and production. The effects of two commercial strains of Trichoderma spp were tested at recommended doses applied to S. aethiopicus. T. harzianum Strain B77 was used as a drench at planting in comparison with a Control and a fungicide in 2005-2006. There were no significant differences between treatments for Harvested Biomass or Survival Percentage. B77 did perform significantly better than the Fungicide in the Total Corm measurement, but neither treatment was significantly different from the Control. In sum, there was a weak trend towards a greater number of output corms as a result of the application of the biocontrol agent. In both 2005-2006 and 2006-2007 we tested T. harzianum Strain kd applied as a drench at planting, with a second drench at 4 weeks. In 2006-2007 there were no significant differences between treatments, but the trend was towards better performance as a result of the drench at planting only. In 2005-2006 open field trials had shown that S. aethiopicus is susceptible to sunburn and Erwinia soft rot when grown in the full sun. Therefore, we tested the effect of various shadecloth densities and colours on production performance in 2006-2007. Treatments were Control (full sun), 40% White (TiO2) (23% shade), 40% Grey (28-30% shade), Light Black (40%), Medium Black (50%), Dark Black (80%), and Red (40%). There were no significant differences between treatments, but the trends indicated that the Control (full sun) and Dark Black (80% shade) performed the worst. Colour of shade did not appear to be important, and plants under all the shadecloths with 40-50% shade grew best. In a factorial trial different levels of Nitrogen, Phosphorous, and Potassium (NPK)were tested, over two seasons. Four levels of each input were used: N at 0 (Control), 40 kg ha-1 (N1), 80 kg ha-1 (N1), and 120 kg ha-1 (N3). P levels were 0 (Control) 60 kg ha-1 (P1) ,120 kg ha-1 (P2) and 200 kg ha-1 (P3). K levels were 0 (Control), 100 kg ha-1 (K1), 200 kg ha-1 (K2), and 400 kg ha-1 (K3). In 2005-2006 there were no significant differences between treatments. In 2006-2007 data there were significant results for Nitrogen only within each repetition. However, significance disappeared when combining across repetitions. We then ran a Bootstrap re-sampling analysis of both 2005-2006 and 2006-2007 data (data were analyzed separately because of different plot sizes and corm numbers in the two years), looking at the optimal level of each macronutrient tested against all combinations of the other two. Though significant results were obtained for each individual level of each macronutrient against the others in combination, the difference between the confidence intervals was not significant. However, there was a clear trend: the optimum N levels were between 40 and 80 kg ha-1; optimum P level was 0 (the Control) and optimum K levels were between 100 and 200 kg ha-1. Tests of handling during harvest, storage, and planting yielded additional useful information for small scale commercial farmers. The optimal harvest time is May, when the false stem and leaves are senescing and yellow, but still upright and visible. Harvest is facilitated by moistening the soil to minimize breaking off of tuberous roots, with simple field washing to remove compacted soil highly recommended. Harvested corms and tuberous roots should be stored under air-restricted, cool conditions because the tuberous roots contain high moisture and will shrivel quickly when left exposed to air, and excessively dried corms will eventually die. Senesced mother corms should be discarded at harvest. Corms are genetically preprogrammed to shoot, so should be planted in September in soft soil, with 1-2 cm of soil coverage. The studies provide a framework for developing the basic agronomy for the domestication and commercial crop production of an endangered medicinal plant species. / Thesis (M.Sc.Agric.)-University of KwaZulu-Natal, Pietermaritzburg, 2011.

The island of Sri Lanka’s exceptional biodiversity and enigmatic biogeography begs investigation, as the island is key in understanding the evolution of the Asian tropical flora. Since the Jurassic, Sri Lanka has been subjected to remarkable tectonic changes, thus its flora could have been influenced by that of a number of nearby landmasses, as well giving Sri Lanka the potential to have played a wider role in the assemblage of floras elsewhere. Firstly, as Sri Lanka originated as a fragment of the supercontinent Gondwana, part of its flora may contain Gondwanan relict lineages. There is also the potential for immigration from Laurasia after the Deccan Plate collided with it 45-50 Mya. Further, Sri Lanka may harbour floristic elements from nearby land masses such as Africa and Southeast Asia as a result of long distance dispersals, and in situ speciation has the potential to have played an important role in enhancing the endemic Sri Lankan flora. I tested the relative contributions of the above hypotheses for the possible origins of the Sri Lankan flora using three representative families, Begoniaceae, Sapotaceae and Zingiberaceae. These families represent both herbaceous and woody elements, and have high diversity across the tropics. Dated molecular phylogenies were constructed for each family. I used recent analytical developments in geographic range evolution modelling and ancestral area reconstruction, incorporating a parameter J to test for founder event speciation. A fine scale area coding was used in order to obtain a better picture of the biogeography of continental Asia. Amongst all the models compared, a dispersal-extinction cladogenesis model incorporating founder event speciation proved to be the best fit for the data for all three families. The dates of origin for Sri Lankan lineages considerably post-date the Gondwanan break up, instead suggesting a geologically more recent entry followed by diversification of endemics within the island. The majority of Sri Lankan lineages have an origin in the Sunda Shelf (53%). Persistence of warm temperate and perhumid climate conditions in southwestern Sri Lanka resembling those of Peninsular Malaysia and Sumatra could have facilitated suitable habitats for these massive dispersals from the Sunda Shelf region. Some trans-oceanic long distance dispersals from Africa (11%) are also evidenced, again these are too young to accept a hypothesis of dispersal during the Deccan Plate’s migration close to the African coast during the late Cretaceous, but occurred later during the Miocene. Further, some lineages of Laurasian origin (20%) are evidenced in the Zingiberaceae with ancestral areas of China and Indochina, which is congruent with a post collision invasion. Among the families tested, dispersals have occurred stochastically, one during the Eocene, six during the Oligocene, seven during the Miocene, two during the Pliocene and one during the Pleistocene. The highest number of dispersals occurred during the Miocene when a warm climate was prevailing during the Miocene thermal maximum. My results confirm that in situ speciation is an important contributor to the Sri Lankan flora. More rapid radiation of endemics has occurred during Pliocene-Pleistocene; two endemics in Begoniaceae, ten endemics in Sapotaceae and ten endemics in Zingiberaceae have evolved in situ during this period. Sri Lanka will have been subjected to expansion and contraction of climatic and vegetation zones within the island during glacial and interglacial periods, potentially resulting in allopatric speciation. As a conclusion, long distance dispersals have played a prominent role in the evolution of the Sri Lankan flora. The young ages challenge the vicariant paradigm for the origin and current disjunct distributions of the world’s tropical lineages and provide strong evidence for a youthful tropics at the species level. The thesis contains six chapters; first two are introductory chapters, then there are three analytical chapters, one for each family, and finally a summary chapter is provided. Each analytical chapter is written as a stand-alone scientific publication, thus there is some repetition of relevant content in each.