Effect of CDC25 and WEE1 on Plant Cell Cycle and Morphogenesis

The cell cycle comprise the four phases of, G1, S-phase, G2 and mitosis. Two critical transitions are G1/S and G2/M; the latter is regulated by WEE1 kinase and CDC25 phosphatases. The scope of this thesis was to investigate the regulation of the G2/M transition of the cell cycle by WEE1 and CDC25, and how these genes interface with plant growth regulators in Arabidopsis thaliana. In Arabidopsis roots, the frequency of lateral roots was found to be increased by ectopic expression of Schizosaccharomyces pombe (Sp)cdc25e and reduced by Arath;WEE1 expression. I examined the effect of Arath;WEE1 and Spcdc25 on induction of shoots and roots in Arabidopsis hypocotyls in vitro. Hypocotyl explants from two over-expressing WEE1 lines , three T-DNA insertion lines and two expressing cdc25 (Spcdc25e) lines together with wild type (WT) were cultured on two-way gradients of kinetin (Kin) and naphthyl acetic acid (NAA). Below a threshold concentration of NAA (100 ng ml-1), WEE1 repressed morphogenesis in vitro, whereas at all NAA/Kin combinations Spcdc25 promoted morphogenesis (particularly root formation) over and above that in WT. Loss of function wee1-1 cultures were very similar to WT. Quantitative data indicated a significant increase in the frequency of root formation in Spcdc25e cultures compared with WT particularly at low Kin concentrations, and WEE1oe’s repressive effect was overcome by NAA but not Kin. In conclusion, WEE1 has a repressive effect on morphogenesis in vitro that can be overcome by auxin whereas Spcd25 by-passes a cytokinin requirement for the induction of morphogenesis in vitro. The role of CDC25 and WEE1 in DNA damage responses was also analysed. Two over-expressing Arath;CDC25 lines and T-DNA mutants showed no difference to WT either in standard conditions or zeocin-supplemented treatments. However, root length was longer in Arath;CDC25oe lines treated with hydroxyurea (HU) and lateral root number was increased compared to WT. This suggests a differential response of Arath;CDC25oe in the DNA replication (HU-induced) and DNA damage (zeocin-induced) checkpoints (Chapter 5). Finally the roles of WEE1 and CDC25 in cell cycle regulation were examined using tobacco TBY-2 cell cultures expressing Arath;WEE1, Nicotiana tabacum (Nicta)WEE1 or Arath;CDC25. Whilst Nicta;WEE1 lengthened G2 of the cell cycle, Arath;WEE1 had an unusual effect of shortening G2 phase and Arath;CDC25 had no observable effect (Chapter 6).

Complex Interplay Between Determinants of Pacing and Performance During 20 km Cycle Time Trials

Purpose This study examined the determinants of pacing strategy and performance during self paced maximal exercise. Methods Eight well trained cyclists completed two 20 km time trials. Power output, RPE, positive and negative affect, and iEMG activity of the active musculature were recorded every 0.5km, confidence in achieving pre-exercise goals was assessed every 5 km, and blood lactate and pH were measured post-exercise. Differences in all parameters were assessed between fastest (FAST) and slowest (SLOW) trials performed. Results Mean power output was significantly higher during the initial 90% of FAST, but not the final 10%, and blood lactate concentration was significantly higher and pH significantly lower following FAST. Mean iEMG activity was significantly higher throughout SLOW. RPE was similar throughout both trials, but participants had significantly more positive affect and less negative affect throughout FAST. Participants grew less confident in their ability to achieve their goals throughout SLOW. Conclusions The results suggest that affect may be the primary psychological regulator of pacing strategy and that higher levels of positivity and lower levels of negativity may have been associated with a more aggressive strategy during FAST. Although the exact mechanisms through which affect acts to influence performance are unclear, it may determine the degree of physiological disruption that can be tolerated, or be reflective of peripheral physiological status in relation to the still to be completed exercise task.

Mental Toughness Moderates Social Loafing in Cycle Time-Trial Performance

Purpose: The purpose of this study was to determine if mental toughness moderated the occurrence of social loafing in cycle time-trial performance. Method: Twenty-seven men (Mage = 17.7 years, SD = 0.6) completed the Sport Mental Toughness Questionnaire prior to completing a 1-min cycling trial under 2 conditions: once with individual performance identified, and once in a group with individual performance not identified. Using a median split of the mental toughness index, participants were divided into high and low mental toughness groups. Cycling distance was compared using a 2 (trial) × 2 (high–low mental toughness) analysis of variance. We hypothesized that mentally tough participants would perform equally well under both conditions (i.e., no indication of social loafing) compared with low mentally tough participants, who would perform less well when their individual performance was not identifiable (i.e., demonstrating the anticipated social loafing effect). Results: The high mental toughness group demonstrated consistent performance across both conditions, while the low mental toughness group reduced their effort in the non-individually identifiable team condition. Conclusions: The results confirm that (a) clearly identifying individual effort/performance is an important situational variable that may impact team performance and (b) higher perceived mental toughness has the ability to negate the tendency to loaf.