A subset of mitochondrial group II introns of flowering plants has, over evolutionary time, lost characteristic features and employs unconventional splicing pathways. Given the potential impact of cold treatment on RNA folding, as well as on enzymatic activity and import of nuclear-encoded splicing machinery, I have examined the physical excised forms of aberrant introns from wheat embryos subjected to 4oC. My findings suggest a shift in biochemistry with cold treatment to novel splicing pathways that generate heterogeneous in vivo circularized forms for nad1 intron 2, nad2 intron 1 and the cox2 intron, in contrast to predominantly linear excised intron forms at room temperature. Interestingly, the highly degenerate nad1 intron 1, which due to DNA rearrangement has been broken into two halves that interact for splicing in trans, is excised exclusively by first-step hydrolysis at room temperature and under cold treatment. In this case, splicing culminates in two distinct linear half introns that appears correlated with an unusual 5’ terminal insert. This represents the first in vivo demonstration of hydrolytic trans-splicing. Based on northern analysis, cold treatment was further associated with reduced splicing efficiency for all introns surveyed. Moreover, study of precursor transcripts of the nad1a-intron 1a locus suggests the efficiency of end-maturation, including processing of the cotranscribed tRNA-Pro gene, is also reduced in the cold. My findings demonstrate a temperature-sensitivity of transcript maturation, particularly for RNA splicing, providing new insight into the impact of cold growth conditions on plant mitochondrial gene expression.

In this thesis seafloor cold vents are examined using autonomous underwater vehicle (AUV) and remotely operated vehicle (ROV) data on the Northern Cascadia margin. These data were collected in a 2009 joint cruise between the Monterey Bay Aquarium Research Institute (MBARI) and Natural Resources Canada (NRCan). High- resolution bathymetry data, acoustic reflectivity (backscatter) data, and 3.5 kHz sub bottom profiler data were examined for cold-vent-related features that include pockmarks, chemosynthetic biological communities (CBC), and authigenic carbonate. Additionally subsequent ROV observations, sediments from push cores and seafloor video/photos were used to ground truth AUV data. Numerous prolific venting sites were examined in detail and a model for the evolution of venting was generated. Vents are categorized as juvenile, intermediate, or mature depending on the presence and or absence of cold-vent-features. High near-surface reflection amplitudes are coincident with an anomalous area of seafloor backscatter. In June of 2012, NEPTUNE (North East Pacific Time-series Underwater Networked Experiment) collected a near-surface push core with their ROV ROPOS (Remotely Operated Platform for Ocean Sciences) in the high reflective area. The retrieved core showed stacked turbidites in the top 0.5 meters of the sediment column. Closely spaced high-velocity turbidite sands are highly reflective and inhibit acoustic penetration to depth. The presence of high-density, high-velocity sands in the near surface is linked to steady ocean bottom currents. These bottom currents progress northeast to southwest over the study area and differentially erode the surface sediments by removing muds and leaving heavy sands over the exposed area. / Graduate / 0373 / 0374 / jonfurlong@hotmail.com

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Frost assays carried out on winter barley (Hordeum vulgare cv Igri) showed that a single (10ml I) application of liquid seaweed extract (LSE) marginally increased the frost resistance of non-acclimated (NA) plants by 2.3% compared to NA controls and cold-acclimated (CA) plants by 2.1% compared to CA controls. Three applications of LSE increased the frost resistance of NA plants by 16% compared to NA controls and CA plants by 7.5% compared to CA controls. These observations were durable in a small scale field trial where LSE increased plant dry weights (control 0.55, single LSE, 0.611 and multiple LSE 0.621 log dry weight), but rain following LSE application reduced LSE mediated frost resistance. Glasshouse growth trials illustrated that LSE enhanced tiller production (control 2.8, one LSE 3.8 and three LSE 4.5 tillers) and dry weight gain, but where precipitation followed LSE application, up to 3 days post application, the LSE mediated effect was not sustained. Protein analysis demonstrated that cold-acclimation and LSE treatments increased the total soluble protein content of winter barley. A single application of LSE increased the soluble protein content of NA plants by 36.7% and three applications of LSE to NA plants increased protein concentration by 86.5%. There was not a significant increase in the soluble protein concentration of LSE treated CA plants. There was a significant increase in the number of high molecular weight proteins and the up-regulation of a 118kDa and a 57kDa protein when plants were treated with LSE. However precipitation following LSE application adversely affected LSE mediated protein expression. A tentative immunological identification of the up-regulated proteins suggested that the 118kDa protein is a dehydrin. There was a 2 fold decrease in plant water potential of NA plants treated with three applications of LSE compared to controls and a similar decrease in plant water potential was observed in cold-acclimated plants. The duration of LSE mediated decline in water potential lasted for 6 days, post LSE application. However there was no significant reduction in the percentage water content of cold-acclimated and LSE treated plants. Differential scanning calorimetry demonstrated that both cold-acclimated and LSE treated plants had significantly less frozen water in their crown tissue compared to non-acclimated controls. Further thermal analysis (infrared thermography and thermocouple data) showed that both cold-acclimation and LSE treatments reduced the speed of water removal from plant cells to the extracellular ice (NA 4.06, NA3LSE 13.4, CA 15.7 and CA3LSE 19.31 minutes). It is hypothesised that both CA and LSE treatments are modifying plant water status, so that water becomes more structured at the physico-chemical level, and thus alters the osmotic behaviour of cellular water. This higher level of water structuring reduces frost damage by conserving the cellular water environment and thus reducing protein denaturation and membrane damage.

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It has been demonstrated that EGCG and caffeine, naturally present in green tea, have thermogenic properties in thermoneutral conditions. The purpose of this study was to quantify the effect of the combined ingestion of EGCG/caffeine on thermogenic responses during a 3h mild cold exposure. Eight healthy males (22± 1 y) were exposed in a randomized, cross over, single blinded fashion to the cold (liquid conditioned suit perfused with 15°C water), after ingesting either a placebo (CON) or an extract of 1600mg of EGCG and 600mg of caffeine (EXP). Thermic, metabolic and electromyographic measurements were monitored at baseline and during cold exposure. After 180min of cold exposure, shivering intensity was significantly reduced by ~32% in EXP condition compared to CON. Area under the curve calculations for total shivering intensity was also reduced by ~21% in EXP (457±99 %MVC.min) compared to CON (361±81 %MVC.min; p=0.007). In contrast, the total area under curve of VO2 was ~25% higher in EXP (33.3±5.5 L O2) compared to CON (25.3±5.1 L O2; p=0.03). Total Heat production (Hprod) also increased by about 11% in the EXP condition (1535±112 kJ) compared to control (1372 ±106 kJ; p=0.002). The decrease in shivering activity combined with an increase in VO2 and Hprod, following the ingestion of EGCG and caffeine in the cold, indicates that nonshivering thermogenesis pathways can be significantly stimulated in adult humans.

Mycorrhiza is an association between a host plant and a soil fungus. Experiments were conducted to determine low temperature and soil disturbance effects on AM fungus winter survival and vigour in spring. The results showed that cool temperatures significantly reduced plant root growth and delayed AM formation. Glomus intraradices sporulation was highest at 23°C, while spore metabolic activity was significantly reduced with temperature below 10°C. Root length and colonization percentage decreased at 10°C. Mycorrhizal fungi increased 32P activity of leek leaves at a root zone temperature of 23°C 7 days after 32P injection, and at both 23°C and 15°C 14 days after injection. No difference was found at 0°C between mycorrhizal and non-mycorrhizal plants. Amounts of total and metabolically active spores and hyphae varied over sampling times. The infectivity of AM fungi was not affected by soil disturbance, but varied among the sampling times.

The nutritional value of biodiesel co-products were studied for swine. In Exp. 1, expeller-pressed canola meal was nutritionally characterized and validated for grower-finisher pigs. Expeller-pressed canola meal provided adequate energy and AA; ADG was reduced 3 g/d per 1% expeller-pressed canola meal inclusion in diets formulated to equal NE and SID AA, due to 5 mol/g dietary glucosinolates. In Exp. 2, cold-pressed canola cake samples from 4 different processing conditions were tested against expeller-pressed canola meal and seed in a digestibility study. Higher residual oil in the cake increased the DE and NE content. In Exp. 3, 15% of either solvent-extracted or expeller-pressed canola meal with or without 5% glycerol was tested against a soybean meal diet for weanlings. Solvent-extracted or expeller-pressed canola meal, or in combination with 5% glycerol can partially replace soybean meal in weaner diets formulated to equal NE and SID AA content without affecting growth performance. / Animal Science