UBC Theses and Dissertations
Functional genomics of plant chitinase-like genes Johnston, David Morris
The Arabidopsis chitinase-like1 (Atctl1) mutant, pom1 is compromised in primary cell wall development, resulting in short roots when grown on high sucrose and shortened hypocotyls when grown in darkness. To better understand this phenotype and the evolution of AtCTL1 and its homologue, AtCTL2, we obtained a large number of CTL sequences and determined the phylogenetic relationships among them. Since microarray analysis had suggested a change in auxin response or homeostasis in pom1, I used the auxin reporter DR5::GUS in the pom1 background to assess changes in distribution. To assess whether the biochemical functions of AtCTL1 homologues in Arabidopsis and other plants are conserved, I transformed pom1 with AtCTL2 and CTLs from poplar (Populus trichocarpa x Populus deltoides clone H-11) and from Picea glauca (spruce) and assessed rescue of the pom1 phenotype. To further understand CTL expression and function, Arabidopsis and poplar CTL promoter::GUS fusions were also expressed in Arabidopsis, PopCTL1 overexpressed in Arabidopsis, and CTL expression down regulated in poplar by RNAi. Our results indicate that CTL genes represent an ancient family encoding proteins of conserved biochemical function. In dicots, represented by Arabidopsis and poplar) duplicated CTL genes are differentially expressed in conjunction with primary and secondary cell wall development, respectively. Mutation of these genes results in improperly formed primary walls in certain cell types in the case of AtCTL1, and an impairment in the differentiation of vascular bundles for AtCTL2. Overexpression of PopCTL1 in Arabidopsis seems to over stimulate the differentiation of vascular bundles, and our studies show that auxin distribution is altered in the Atctl1 mutant. Down regulation of PopCTL1 and PopCTL2 in poplar appears to phenocopy aspects of these mutations, resulting in secondary cell walls that appear to have less deposition of lignin and an accelerated production of secondary xylem respectively. While specific biochemical function(s) of CTL genes were not studied, potential functions are discussed.
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