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Common gromwell (Lithospermum officinale) acts as a host plant for the monophagous moth Ethmia dodecea  whose larvae are completely dependent on the leaves. As conservation authorities now want to reinstate the regionally extinct moth to Mälardalen, a stable population of its host plant is a requirement. To facilitate the work of strengthening the presence of gromwell a partnership was therefore initiated between Västmanland County Board and Uppsala University. In this cooperation, I performed two studies. In the first one I examined how water and temperature affect plant germination and how nutrient levels affect early growth. In the second study I investigated whether the germination is influenced by chemical treatment (soaking in acid) and mechanical damage (seeds scratched with sandpaper) on the seeds. I worked with the hypothesis that gromwell is grazed by cows and is therefore dispersed and germinates in the spring. This should mean high water levels combined with high temperatures would produce higher germination. For the second study, it means that the germination rate should be higher in the seeds treated with the acid than in the scratched and control treatments. In the first study, so few seeds germinated that I could not draw any conclusions, but germinations appear to go faster in the combination with high nutrients high temperature and frequent watering. In the second study, the seeds scraped with sandpaper had the highest germination rate. This indicates that gromwell may be dispersed by birds, and I propose sandpaper rubbing as a method to easily increase the germination rates of L. officinale in greenhouses in order to reinforce small populations in the field.

With the ever increasing reliance on hydropower in the world, it is important that its effects on riparian vegetation is well understood. This study aimed to explore the effects of different forms of hydropeaking, which results from the rapid and frequent opening and closure of hydroturbines, on different species groups of riparian species native to northern Sweden. Thirteen riparian species belonging to three different species groups, forbs, graminoids and woody species, were subjected to three different watering treatments simulating water stress, rapid fluctuation and prolonged flooding, which often correspond to different hydropeaking conditions. A fourth treatment maintained individuals under controlled conditions and simulated a non-regulated stream. The treatments were carried out over seven weeks. The results showed that the treatments affect the three tested species groups differently. Woody species responded to all treatments with lower germination, higher mortality and lower root elongation, while forbs and graminoids responded to flooding and fluctuation with increased germination but higher mortality and lower root elongation. Fluctuation and flooding had in general positive effects on germination and short term growth but also increased mortality, particularly fluctuation. There were no significant results regarding change in number of leaves for any treatment or species group. As this study only covered seven weeks it can only account for short-term trends, but it still provides interesting information about general expected trends for a wide variety of species native to northern Sweden and belonging to the main three species groups. Such information is key for river management.

Free radical and aldehydic breakdown product content were determined, by EPR and UV / visible spectroscopy, primarily in intermediate (desiccation tolerant) seeds of <i>Carica papaya</i> L. (Papaya) and recalcitrant (desiccation intolerant) seeds of <i>Aesculus hippocastanum</i> L. (Horse chestnut), but also in other species covering a range of desiccation tolerances, with a view to determining the role of oxidative stress as a diagnostic marker for desiccation tolerance. Axes of non-senescent highly viable recalcitrant seeds of horse chestnut were metabolically active, contained products of lipid peroxidation, displayed low levels of enzymatic protection against activated oxygen and peroxides, and a two-peak free radical EPR signal. During fully hydrated storage at 16 °C for up to 18 months, seeds exhibited, sequentially, an increase in germination rate, a transient increase in intensities of both the low field and high field EPR peaks, a significant increase in membrane leakage and decrease in seed viability, germination rate, and SOD and peroxidase activities. Drying 'unstored' seeds below and embryonic axis moisture content of 40 to 50 % initiated viability loss. At < 25 % moisture content all axes were inviable and displayed a 2- to 4-fold increase in solute leakage, lipid peroxidation products and the low field EPR signal. Seed desiccation sensitivity increased with hydrated storage. The accumulation of lipid peroxidation products and free radicals on drying generally occurred to a greater extent, or at a higher moisture content, than observed with unstored seeds. The results indicate a mediating role for oxidative stress in recalcitrant seed viability loss which is differentially expressed during hydrated, 'natural' ageing and desiccation. Similar trends were seen in other recalcitrant species with the increase in lipid peroxidation products occurring around the point of viability loss. However the study of a more orthodox species (papaya) revealed no such trends.

Bacillus anthracis has been classified as one of the most dangerous bioterrorism agents causing high mortality rates in short periods of time. Anthrax spores are extremely resistant to chemical and environmental factors, and have the ability to return into a vegetative (virulent) state during the process of germination. Previous research has suggested that spores can be eradicated with common disinfectants after germination and release of spore coats. During germination, the spore coat is degraded, making the spore susceptible to penetration of chemicals into the spore core. While previous research has focused on a qualitative understanding of germination of spores by obtaining high-resolutions images of spore coats to understand how protein coat layers change during germination, very few studies have evaluated changes in mechanical properties of spores during germination, and how germination affects virulence of macrophages. In this study, we performed a series of in vitro experiments to do an in-depth analysis of germination and virulence of B. anthracis. Atomic force microscopy (AFM) was used to investigate changes in spore surface properties during germination including morphology, roughness, elasticity, and spring constant. AFM results suggested that germination mechanisms depend on germinants used to trigger germination and roughness of Bacillus species increase during germination. In addition, the elasticity and spring cell constant of B. anthracis spores are affected during germination since the elastic moduli and cell spring constant values decreased with time as the spore was germinating, making the cells more susceptible. Spore killing was also tested both in sporulated and vegetative B. anthracis using the antimicrobial peptide chrysophsin-3 and the surfactant dodecylamine (DDA). Both killing agents were capable of eradicating B. anthracis spores, but more killing was observed for spores that were germinating or had become vegetative. The presence of germinant receptors from the Ger operon and its role on germination kinetics of B. anthracis was also investigated. The germination of mutant spores that carried one receptor or lacked all germinant receptors was compared to the germination kinetics of wild-type B. anthracis. Our results suggest that germination of spores is modified by the presence or absence of germinant receptors. Furthermore, the mutant B. anthracis strain lacking all receptors germinated suggesting that other receptor independent pathways may exist in B. anthracis. Finally the ability of B. anthracis to adhere, grow, and invade macrophages was investigated. Invasion of macrophages by B. anthracis was dependent on germinant receptors and the ability of spores to germinate and multiply. Our results suggest that macrophages were not capable of killing infecting spores, and on the contrary, germination of spores inside macrophages caused the lysis of macrophages. An uncontrolled release of cytokines by macrophages was elicited by spores and germinated B. anthracis. Our study helps understand the process of germination of B. anthracis spores at a nanomolecular level. Our investigation may be a valuable tool in the design and development of antisporal compounds.

Typescript (photocopy). / Digitized by Kansas Correctional Industries

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Wang, Hao. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 83-93). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

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