Recent computational analysis of expressed sequence tags and expression analysis in plants revealed a variety of non-coding RNAs that are transcribed but not translated to functional proteins. Non-coding RNAs are classified into three groups; small op...
Recent computational analysis of expressed sequence tags and expression analysis in plants revealed a variety of non-coding RNAs that are transcribed but not translated to functional proteins. Non-coding RNAs are classified into three groups; small open reading frame RNAs (sORF RNAs), microRNAs (miRNAs), and short-interfering RNAs (siRNAs). Small ORF RNAs are several hundreds to thousands nucleotides-long mRNAs that contain several coding regions producing less than 100 amino acid-long peptides. miRNAs and siRNAs are 20 to 25 nucleotides-long RNA transcripts that have an ability to bind complementary mRNAs, thereby suppress target gene expression by either mRNA cleavage or translational repression. Although many recent reports propose that non-coding RNAs play diverse roles in growth, development, morphogenesis, and stress responses of plants, the biological roles of different types of non-coding RNAs remain to be elucidated. In this thesis, one sORF RNA designated AtGUT15 and one miRNA designated miRNA417 were investigated for their tissue-specific and stress-responsive expression patterns and the functional roles in seed germination and seedling growth of Arabidopsis thaliana under normal growth or various abiotic stress conditions.
AtGUT15 and miRNA417 are expressed through all growth stages and ubiquitously in all organs including stems, roots, leaves, and flowers. The expression of AtGUT15 and miRNA417 was moderately regulated by cold, drought, or salt stress and by ABA treatment. Loss-of-function mutants of AtGUT15 displayed a much faster bolting compared to wild-type plants under normal growth conditions, implying that AtGUT15 may be involved in regulating bolting time in Arabidopsis plants. To prove the biological roles of miRNA417, transgenic Arabidopsis plants that constitutively overexpress miRNA417 under the control of cauliflower mosaic virus 35S promoter were generated and their phenotypes were analyzed under normal and stress conditions. Results showed that overexpression of miRNA417 had no effect on growth and development of Arabidopsis plants under normal growth conditions. However, seed germination and seedling growth of the transgenic plants were retarded compared with the wild-type plants in the presence of high salt or ABA. These results imply that miRNA417 plays a role as a negative regulator of seed germination in Arabidopsis plants under salt stress. No cleavage of the putative targets for miRNA417 was observed by the overexpression of miRNA417 in the transgenic plants, suggesting that miRNA417 exerts its role by translational suppression mechanism.