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).

Mapping of Submerged Aquatic Vegetation in Rivers From Very High Resolution Image Data, Using Object Based Image Analysis Combined with Expert Knowledge

The use of remote sensing for monitoring of submerged aquatic vegetation (SAV) in fluvial environments has been limited by the spatial and spectral resolution of available image data. The absorption of light in water also complicates the use of common image analysis methods. This paper presents the results of a study that uses very high resolution (VHR) image data, collected with a Near Infrared sensitive DSLR camera, to map the distribution of SAV species for three sites along the Desselse Nete, a lowland river in Flanders, Belgium. Plant species, including Ranunculus aquatilis L., Callitriche obtusangula Le Gall, Potamogeton natans L., Sparganium emersum L. and Potamogeton crispus L., were classified from the data using Object-Based Image Analysis (OBIA) and expert knowledge. A classification rule set based on a combination of both spectral and structural image variation (e.g. texture and shape) was developed for images from two sites. A comparison of the classifications with manually delineated ground truth maps resulted for both sites in 61% overall accuracy. Application of the rule set to a third validation image, resulted in 53% overall accuracy. These consistent results show promise for species level mapping in such biodiverse environments, but also prompt a discussion on assessment of classification accuracy.