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The homeodomain leucine zipper (HD-Zip) genes encode transcription factors that have diverse functions in plant development and have often been implicated in stress adaptation. The HD-Zip genes are the most abundant group of homeobox (HB) genes in plants and do not occur in other eukaryotes. This paper describes the complete annotation of the HD-Zip families I, II and III from rice and compares these gene families with Arabidopsis in a phylogeny reconstruction. Orthologous pairs of rice and Arabidopsis HD-Zip genes were predicted based on neighbour joining and maximum parsimony (MP) trees with support of conserved intron-exon organization. Additionally, a number of HD-Zip genes appeared to be unique to rice. Searching of EST and cDNA databases and expression analysis using RT-PCR showed that 30 out of 31 predicted rice HD-Zip genes are expressed. Most HD-Zip genes were broadly expressed in mature plants and seedlings, but others showed more organ specific patterns. Like in Arabidopsis and other dicots, a subset of the rice HD-Zip I and II genes was found to be regulated by drought stress. We identified both drought-induced and drought-repressed HD-Zip genes and demonstrate that these genes are differentially regulated in drought-sensitive versus drought-tolerant rice cultivars. The drought-repressed HD-Zip family I gene, Oshox4, was selected for promoter-GUS analysis, showing that drought-responsiveness of Oshox4 is controlled by the promoter and that Oshox4 expression is predominantly vascular-specific. Loss-of-function analysis of Oshox4 revealed no specific phenotype, but overexpression analysis suggested a role for Oshox4 in elongation and maturation processes.

A key question in developmental biology is how cells exchange positional information for proper patterning during organ development. In plant roots the radial tissue organization is highly conserved with a central vascular cylinder in which two water conducting cell types, protoxylem and metaxylem, are patterned centripetally. We show that this patterning occurs through crosstalk between the vascular cylinder and the surrounding endodermis mediated by cell-to-cell movement of a transcription factor in one direction and microRNAs in the other. SHORT ROOT, produced in the vascular cylinder, moves into the endodermis to activate SCARECROW. Together these transcription factors activate MIR165a and MIR166b. Endodermally produced microRNA165/6 then acts to degrade its target mRNAs encoding class III homeodomain-leucine zipper transcription factors in the endodermis and stele periphery. The resulting differential distribution of target mRNA in the vascular cylinder determines xylem cell types in a dosage-dependent manner.

The tRNA^Leu (UAA) intron has been recorded in the plastid genome of many algae and land plants and was the first intron to be discovered in cyanobacteria. In all known cases it interrupts the tRNA^Leu anticodon loop at a conserved position (U-intron-AA). Cyanobacteria are a diverse group of photosynthetic prokaryotes, some involved in symbiotic associations with a wide range of organisms. The most studied associations are those with plants, where strains of Nostoc are the common cyanobacterial partner. In this thesis two aspects of the biology of the cyanobacterial tRNA^Leu (UAA) intron are focused: first, the use of the intron as a genetic marker for studying the diversity and specificity of two cyanobacterial symbiosis (bryophytes and cycads) and second, the evolutionary patterns of the intron by using the unique data set generated from the diversity analysis.

From the studies, many different Nostoc strains are involved in the two symbiotic associations, although no variation was observed within a single bryophyte cavity or cycad coralloid root. Furthermore, a certain level of temporal stability in the cyanobiont composition of the bryophyte population was found and, in the cycad association different coralloid roots from a single specimen may harbor different cyanobacteria. That a minor cyanobiont could have avoided detection is still possible but unlikely. The sequence alignment of the Nostoc tRNA^Leu (UAA) introns reveals great sequence similarity with size variation only found in the structural element P6b. This element was found to consist of heptanucleotide repeats and of other non-repetitive genetic elements (NIS elements). The sporadic occurrence of the NIS elements indicates recent origins and a mechanism for its dispersal is proposed.

In this thesis new insights are given concerning cyanobacterial symbioses and also on the mechanisms involved in the evolution of an old genetic element: the tRNA^Leu (UAA) intron in cyanobacteria.

The genertic diversity of cyanobacteria associated with cycads was examined using the tRNA Leu (UAA) intron as a genetic marker. Coralloid roots of both natural populations of the cycad Macrozamia riedlei (Fischer ex Gaudichaud-Beaupré) C.A. Gardner growing in Perth, Australia and cycads growing in greenhouses, also in Perth, were used and their respective cyanobionts analyzed. Several Nostoc strains were found to be involved in this symbiosis, both in natural populations and greenhouse-orginated cycads. However, only one strain was present in individual coralloid roots and in individual plants, even when analyzing different coralloid roots from the same plant. Moreover, when examining plants growing close to each other (female plants and their respective offspring) the same cyanobacterium was consistently present in the different coralloid roots. Whether this reflects a selective mechanism or merely the availability of Nostoc strains remains to be ascertained. The high cyanobacterial diversity in coralloid roots of cycads revealed by PCR fingerprinting is, therefore, contested. In this study, the potential probems of using different methods (e.g. PCR fingerprinting) to study the genetic diversity of symbiotic cyanobacteria, is also addressed.

Homeodomain leucine zipper (HDZip) proteins constitute a large family of transcription factors characterised by the presence of a DNA-binding homeodomain and an adjacent leucine zipper motif, which mediates protein-dimer formation. The HDZip genes of Arabidopsis have been divided into four classes, HDZip I-IV. This thesis describes the characterisation and phylogeny of the class I HDZip genes and focuses on the expression and function of four HDZip I genes, ATHB5, -6, -7 and -16.

The phylogenetic analyses of the 17 HDZip I sequences defined six subclasses, supported by the intron patterning and the traced duplication history of the genes. The members within each subclass showed diversification in expression, suggesting that the cis regulatory regions of the closely related genes have undergone evolutionary changes. However, similarities in the gene expression patterns between genes also exist and external factors like the availability of water and quality of light directs the expression of a subset of HDZip I genes. Expression analyses revealed that the plant hormone abscisic acid (ABA) is involved as a systemic signal for the salt or osmoticum induced ATHB7 expression, whereas light signals mediated through the blue light photoreceptors was found to direct the expression of ATHB6.

Phenotypic analyses of plants with altered levels of ATHB6 or ATHB16 suggested that these paralogous genes encode proteins with very similar functions. ATHB16 was shown to act as a negative regulator of leaf cell expansion, as a suppressor of the flowering time sensitivity to photoperiod and as a positive regulator of blue light dependent inhibition of hypocotyl growth. A similar role for ATHB6 in the regulation of hypocotyl elongation was recorded. Further, analyses of multiple loss-of-function plants demonstrated that ATHB5, -6 and -16 function at least in part redundantly in mediating light effects on hypocotyl elongation.