Functional characterization of two plant type I MADS-box genes in Arabidopsis thaliana : AGL40 and AGL62 : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Plant Biology at Massey University, Palmerston North, New Zealand
MADS-box transcription factors (TF) are a family of evolutionary conserved genes found across various eukaryotic species. Characterized by the conserved DNA binding MADS-box domain. MADS-box TF has been shown to play various roles in developmental processes. MADS-box genes can be based on MADS-box structural motifs divided into type I and type II lineages. In plants very limited functional characterization have been achieved with type I genes MADS-box genes. In this project we attempted to functionally characterize 2 closely related members of the type I lineage MADS-box genes AGL40 and AGL62 and give further support to the hypothesis that plant type I MADS-box genes are also crucial to normal plant development. Based on our expression domain characterization assay using AGL62: GUS fusion construct, we have shown expression of AGL62 in various tissues but especially strong in developing seeds, pollen and seedling roots and shoots. The web based microarray data suggesting that AGL62 may have a function in seed, pollen and seedling development backed up this result. Interestingly when we carried out PCR based genotyping with segregating population of heterozygous AGL62 T-DNA insertion lines (agl62/+) to identify the homozygous T-DNA insertion lines we detected no homozygous T-DNA insertion line indicating loss-of-function of AGL62 may be lethal to plant. With reference to the AGL62 expression in pollen, seed and seedling root and shoot, we carried out phenotypic assay on each of these tissues in agl62/+ background to investigate whether there was any phenotypic defect observed. Significant reduction in number of seeds was observed in agl62/+ indicating possible role of AGL62 in seed development. Our microscopic observation of seeds from agl62/+ plants showed defective embryos and confirmed that AGL62 plays a role in seed development. Our data on AGL62 is the first report that confirms AGL62's involvement in plant development and can be a ground work for further works on functional characterization of other members of plant type I MADS-box genes.