A multi-scale assessment of physiological processes in arctic tundra plants under natural and simulated climate change scenarios

Climate warming is predicted to cause an increase in the growing season by as much as 30% for regions of the arctic tundra. This will have a significant effect on the physiological activity of the vascular plant species and the ecosystem as a whole. The need to understand the possible physiological change within this ecosystem is confounded by the fact that research in this extreme environment has been limited to periods when conditions are most favorable, mid June–mid August. This study attempted to develop the most comprehensive understanding to date of the physiological activity of seven tundra plant species in the Alaskan Arctic under natural and lengthened growing season conditions. Four interrelated lines of research, scaling from cellular signals to ecosystem processes, set the foundation for this study. ^ I established an experiment looking at the physiological response of arctic sedges to soil temperature stress with emphasis on the role of the hormone abscisic acid (ABA). A manipulation was also developed where the growing season was lengthened and soils were warmed in an attempt to determine the maximum physiological capacity of these seven vascular species. Additionally, the physiological capacities of four evergreens were tested in the subnivean environment along with the potential role anthocyanins play in their activity. The measurements were scaled up to determine the physiological role of these evergreens in maintaining ecosystem carbon fluxes. ^ These studies determined that soil temperature differentials significantly affect vascular plant physiology. ABA appears to be a physiological modifier that limits stomatal processes when root temperatures are low. Photosynthetic capacity was limited by internal plant physiological mechanisms in the face of a lengthened growing season. Therefore shifts in ecosystem carbon dynamics are driven by changes in species composition and biomass production on a per/unit area basis. These studies also found that changes in soil temperatures will have a greater effect of physiological processes than would the same magnitude of change in air temperature. The subnivean environment exhibits conditions that are favorable for photosynthetic activity in evergreen species. These measurements when scaled to the ecosystem have a significant role in limiting the system's carbon source capacity. ^

Regulation of Rab5 GTPase activity during Pseudomonas aeruginosa-macrophage interaction

Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen. Several antibiotic resistant strains of P. aeruginosa are commonly found as secondary infection in immune-compromised patients leaving significant mortality and healthcare cost. Pseudomonas aeruginosa successfully avoids the process of phagocytosis, the first line of host defense, by secreting several toxic effectors. Effectors produced from P. aeruginosa Type III secretion system are critical molecules required to disrupt mammalian cell signaling and holds particular interest to the scientists studying host-pathogen interaction. Exoenzyme S (ExoS) is a bi-functional Type III effector that ADP-ribosylates several intracellular Ras (Rat sarcoma) and Rab (Response to abscisic acid) small GTPases in targeted host cells. The Rab5 protein acts as a rate limiting protein during phagocytosis by switching from a GDP- bound inactive form to a GTP-bound active form. Activation and inactivation of Rab5 protein is regulated by several Rab5-GAPs (GTPase Activating Proteins) and Rab5-GEFs (Rab5-Guanine nucleotide Exchange Factors). Some pathogenic bacteria have shown affinity for Rab proteins during infection and make their way inside the cell. This dissertation demonstrated that Rab5 plays a critical role during early steps of P. aeruginosa invasion in J774-Eclone macrophages. It was found that live, but not heat inactivated, P. aeruginosa inhibited phagocytosis that occurred in conjunction with down-regulation of Rab5 activity. Inactivation of Rab5 was dependent on ExoS ADP-ribosyltransferase activity, and more than one arginine sites in Rab5 are possible targets for ADP-ribosylation modification. However, the expression of Rin1, but not other Rab5GEFs (Rabex-5 and Rap6) reversed this down-regulation of Rab5 in vivo. Further studies revealed that the C-terminus of Rin1 carrying Rin1:Vps9 and Rin1:RA domains are required for optimal Rab5 activation in conjunction with active Ras. These observations demonstrate a novel mechanism of Rab5 targeting to phagosome via Rin1 during the phagocytosis of P. aeruginosa. The second part of this dissertation investigated antimicrobial activities of Dehydroleucodine (DhL), a secondary metabolite from Artemisia douglasiana, against P. aeruginosa growth and virulence. Populations of several P. aeruginosa strains were completely susceptible to DhL at a concentration between 0.48~0.96 mg/ml and treatment at a threshold concentration (0.12 mg/ml) inhibited growth and many virulent activities without damaging the integrity of the cell suggesting anti-Pseudomonas activity of DhL.