<p dir="ltr">H3K27me3 is a transcriptional repressive epigenetic mark that plays vital roles in many biological processes in <i>Arabidopsis thaliana</i>. A number of biochemical and functional characterizations of PKL, an ATP-dependent CHD chromatin remodeler, suggest that PKL contributes to maintain the homeostasis of H3K27me3. To identify other factors that act with PKL together to contribute to the homeostasis of H3K27me3, we undertook an EMS-mutagenesis screen for <i>pkl</i>-associated phenotypes. This genetic screen suggests that PKL may contribute to maintaining the homeostasis of H3K27me3 in an H2A.Z associated or a Mediator associated pathway.</p><p dir="ltr">Here, we took advantage of a combined genetic and bioinformatic method to characterize the contribution of PKL in these two pathways as described above. Our analysis revealed a robust genetic interaction between <i>HTA9</i>, <i>HTA11</i>, and <i>PKL</i> in maintaining proper H2A.Z distribution and enrichment of H3K27me3. In addition, the characterization also sheds light on unexpected roles of PKL in promoting the homeostasis of H3K4me3 and acting with histone demethylases to promote removal of H3K27me3 in an H2A.Z dependent manner. Furthermore, our result also raised the possibility that the tail module of the Mediator complex also plays a critical role in the homeostasis of H3K27me3. While we were examining <i>PKL</i>-dependent chromatin features, we largely optimized the protocol for preparation ChIP-seq samples and libraries and implemented a gene-centric ChIP-seq bioinformatics pipeline for providing robust analysis.</p><p dir="ltr">Ultimately, the work presented in this thesis highlights several divergent pathways that PKL contributes to maintain chromatin homeostasis. By and large, the combined observation from this thesis advances our knowledge of how PKL interacts with other chromatin-associated machineries together to maintain proper epigenetic states and promote other more emergent DNA-templated processes, including replication and transcription.</p>

Early in 1962 the Forest Research Institute of the New Zealand Forest Service became aware that stands of second crop Pinus radiata (D. Don) on some areas of the Moutere Gravel formation were showing slow growth and had a chlorotic appearance (Fig. 1.1). The second crop followed clear felling of mature P. radiata trees and were aged from 0 to 15 years (stone and Will, 1965). It was felt that the apparent reduced growth of the second generation had much in common with similar productivity decline reported especially in European forestry literature. Stone and Will (1965) postulated that the immediate cause o£ the decline was a deficiency of nitrogen highlighted by the low levels of nitrogen in the leaves of the second crop trees, especially those growing on ridge sites. Numerous field trials have been laid out but many of the trials were poorly designed and consequently could not provide statistically sound results. However, some indication of nutrient deficiencies which occur on the Moutere Gravels were obtained. Even before these trials were laid down nutrient deficiencies had been highlighted by early attempts at farming. It was the partial failure of these crops that initially led to the planting of exotic pines, in the belief that these trees thrived on a limited supply of nutrients. The first crop of pines generally fulfilled expectations but nutrient deficiencies started to appear in extensive areas of the second crop. Accordingly, the Forest Research Institute made available three scholarships to study different aspects of the problem. Work was started on a study of the soil sequence across the Moutere Gravels to determine if there was a general decline in fertility of tho soil with the age of the soil and the environmental factors, such as climate which differs in the high inland areas and the low seaside areas of the Moutere Gravels, rather than a particular decline in fertility induced by the first crop of P. radiata. In another study the major weed species Ulex europaeus and Cytisus scoparius was studied to see if its value as a nitrogen fixer would outweigh its disadvantages as a silvicultural weed. Thirdly, a study of the microbial ecology of the soils was undertaken. Whyte (1966) reported that the second rotation trees started to increase their growth rate after approximately five years to a level paralleling the estimated growth rate of the first crop. It was postulated that the residues (needles, roots and branches) remaining after clear felling could cause an increase in microbial numbers and activity with a consequent immobilization of mineral nutrients which were not initially very plentiful. For this reason an area in Tasman Forest was selected in which mature trees and regeneration up to nine years old were found together to study microbial activity and numbers, energy dissipation and nitrogen dynamics to determine if immobilization of nutrients was causing the apparent declines.

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