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Characterization of Complex Systems Using the Design of Experiments Approach: Transient Protein Expression in Tobacco as a Case Study
Published on: January 31, 2014
Two tobacco proline dehydrogenases are differentially regulated and play a role in early plant development
Alexandra Ribarits1, Alisher Abdullaev, Alisher Tashpulatov
1Max F. Perutz Laboratories, University Departments at the Vienna Biocenter, Department of Plant Molecular Biology, University of Vienna, Dr. Bohrgasse 9, 1030 Wien, Austria.
Abstract:
Proline dehydrogenase is the rate-limiting enzyme in proline degradation and serves important functions in the stress responses and development of plants. We isolated two tobacco proline dehydrogenases, NtPDH1 and NtPDH2, in the course of screening for genes upregulated in stressed tobacco (Nicotiana tabacum) microspores. Expression analysis revealed that the two genes are differentially regulated. Under unstressed conditions, their steady-state transcript levels were similar in mature pollen and apical meristems, whereas NtPDH2 was expressed predominantly in vegetative organs, styles, and ovules. The expression of NtPDH1 was maintained at a constant low level during 24 h of dehydration, whereas NtPDH2 was upregulated within 1 h after the onset of stress and subsequently downregulated to undetectable levels. Differential and sustained expression was also found for the two enzymatic isoforms of Arabidopsis thaliana AtPDH. Silencing of the NtPDH genes by RNA interference using the CaMV 35S promoter led to increased proline contents, decreased seed set, delayed seed germination and retarded seedling development pointing towards an important function of at least one of the two NtPDH genes during plant reproductive development.
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Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...
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