Call number: LD2668 .T4 1963 S53 / Master of Science

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"The in vitro addition of plant growth regulators (i.e. phytohormones) to Agrobacterium transformed hairy root cultures affects morphological and biochemical changes, resulting in altered growth and secondary metabolite accumulation rates in root tissues. Significant increases in both growth and secondary product accumulation have been observed, upon incubation with phytohormones, in some species. Consequently, the use of phytohormones in vitro has received increasing attention as a potential means for increasing those plant secondary products notoriously produced in small quantities. However, currently little is known about the specific effects of phytohormones on growth and secondary metabolism. The Chinese herb Artemisia annua L. produces artemisinin, an effective antimalarial therapeutic. Efforts to increase the amount of artemisinin via chemical synthesis or field-grown crops have met with huge costs and disappointingly low yields, respectively. Agrobacterium transformed hairy root cultures of A. annua (Clone YUT16) produce artemisinin and undergo rapid growth compared to non-transformed, making them a good model system to study secondary metabolite production. Demonstrated herein is the first definitive evidence, by any hairy root species, of a favorable response to exogenous combinatorial hormone application as well as the development of a two-stage culture system alluding to optimal growth and artemisinin production conditions in A. annua hairy roots. Furthermore, analysis of artemisinin and biomass accumulation in A. annua hairy roots in the presence of phytohormones has revealed effective individual as well as combinatorial phytohormone concentrations suitable for increasing single and bulk root growth, and artemisinin production. The effectiveness of an optimal phytohormone combination, with respect to time of addition, its relationship to inoculum size, and its combination with the provision of fresh nutrients and or mechanical stress to the roots is also described resulting in artemisinin yields of up to 0.8 ìg/g F.W. Although the findings contained herein are not yet optimized they do, however, argue for the potential usefulness of a two-stage production scheme using phytohormones to increase plant secondary metabolite production in vitro."

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In this study, both physiological and cellular effects are elicited by natriuretic peptides (NPs), a novel type of plant hormone. It was found that rat ANP (rANP) influenced stomatal opening movement in Tradescantia sp., where a significant increase in stomatal opening was observed in the presence of 1 µM rANP. Furthermore, this effect is mediated by cGMP, a (putative) second messenger of NPs. Two inhibitors of guanylyl cyclase, LY 83583 and methylene blue, inhibited rANP-induced stomatal opening. In contrast, stomatal opening is induced in a concentration dependent manner by the cell permeant cGMP analogue 8-Br-cGMP. In addition it was found, that like in animals, the secondary structure of rANP is essential for rANP responses. Linearised rANP is biologically inactive. Since ANP elicit plant responses, an attempt was made to isolate NP analogues from plants. A protocol for partially purifying NP from plants was developed. It was found that two fractions eluted from an immunoaffinity chromatography column (0.5 M KCI eluted fraction and 0.75 M KCI eluted fraction) were biologically active. The level of cGMP in response to NPs was also tested. It is suggested that the receptor of NP is specific since only 0.75 M KCI eluted fractions increased cGMP levels in Zea mays root stele tissue. rANP did not elicit an effect on cGMP levels in this tissue and LY 83583 did not affect this response. It is therefore argued that a plant specific biologically active NP system is present in the stele and it is predicted that NPs modulate solute movement in this tissue. NPs also influence K⁺, Na⁺ and H⁺ fluxes in Zea mays root stele. Increase in both K⁺ and Na+ uptake were observed after 30 min., while H⁺ flux shifted immediately toward influx in the presence of both 0.5 and 0.75 KCI eluted fractions. Finally, a model is proposed for the effect of NPs on solute movement and its signalling system in plants.

Plant growth, development, and environmental responsiveness are dependent on hormone-induced gene expression. This dissertation reports multiple interactions between the plant hormones auxin and ethylene and investigates their contribution to the gravitropic response, elongation growth, adventitious root formation, callus and tracheary element initiation and growth, and flower development. Four mutants of tomato (Lycopersicon esculentum, Mill.) altered in either hormone production or hormone response were used to test the involvement of ethylene and auxin. These mutants included diageotropica (dgt) which is auxin-resistant, Never-ripe (Nr), which is ethylene-resistant, epinastic (epi), which overproduces ethylene and lazy-2 (lz-2), which exhibits a phytochrome-dependent reversed-gravitropic response. Additionally, a double mutant between Nr and dgt was constructed and tested. Gravitropism was studied as an exemplary process involving both auxin and ethylene. Mutant analysis demonstrated that ethylene does not play a primary role in the gravitropic response via the currently known ethylene response pathways. However, ethylene can modify the gravitropic response, e.g. the delayed gravitropic response of the dgt mutant can be restored with exceedingly low concentrations of ethylene and ethylene synthesis- and ethylene-action inhibitors can partially inhibit the graviresponse. The role of gravity in tracheary element (TE) production was tested in microgravity (during a space shuttle flight) and in hypergravity (centrifugation). A correlation was found between gravitational force and the production of TEs, with decreased numbers of TEs produced in microgravity and increased numbers produced in response to hypergravity. Increased production of TEs by dgt in both increased and reduced gravity indicates that gravity regulates vascular development via a DGT-dependent pathway involving auxin. Combination of both the Nr and dgt mutations in a double mutant leads to plants which exhibit the reduction of auxin-sensitivity typical of dgt as well as a delay in fruit ripening typical of Nr. The reduced gravitropic response of the dgt mutant was restored to wild-type levels in the double mutant confirming a complex role for ethylene in the gravitropic response. Abnormal floral organ development was observed in a subset of double mutant flowers.These data demonstrate multiple connections between auxin and ethylene during development and provide further insight into their cellular interactions. / Graduation date: 2001

Thesis (Ph. D.)--Hong Kong University of Science and Technology, 2003. / Includes bibliographical references (leaves 197-236). Also available in electronic version. Access restricted to campus users.

published_or_final_version / Botany / Master / Master of Philosophy