Potato (Solanum tuberosum L.) tubers respond to environmental stresses by alterations of macromolecular synthesis. In an aerobic environment tubers respond rapidly to wounding by synthesizing a set of proteins, the most prominent of which display apparent molecular weights of 78, 48, 38, and 31 kilodaltons. These proteins become intensely labeled by [35Sjmethionine within 2 hours of wounding. The 78 kilodalton polypeptide has been identified by immunoprecipitation as phenylalanine ammonia-lyase. By contrast, tubers incubated in hypoxic conditions for a period as short as 1.5 hours exhibit significantly reduced incorporation of amino acids such that newly synthesized polypeptides are not detected. However, a second set of proteins is synthesized by wounded tubers after prolonged incubation in a hypoxic environment. One peptide of this set is precipitated by an antibody directed against aldolase; several of these proteins may be enzymes of glycolysis necessary for anaerobic metabolism. The results indicate that there is a complex regulatory mechanism which allows mature potato tubers to respond to changes in the environment.
In response to salinity or drought stress, the facultative halophyte Mesembryanthemum crystallinum will switch from C3 photosynthesis to Crassulacean acid metabolism (CAM). During this switch, the transcription rates of many genes encoding glycolytic, gluconeoagenic, and malate metabolism enzymes are increased. In particular, transcription of the Ppc1 and Gap1 genes encoding a CAM-specific isozyme of phosphoenolpyruvate carboxylase and NAD-dependent glyceraldehyde-3-phosphate dehydrogenase, respectively, is increased by salinity stress. To investigate the molecular basis of salt-induced gene regulation, we examined the Ppc1 and Gap1 promoters for cis-elements and trans-acting factors that may participate in their expression. Ppc1 or Gap1 promoter-beta-glucuronidase chimeric gene constructs containing various deletions were introduced into intact, detached M. crystallinum leaves by microprojectile bombardmen. The Ppc1 5'-flanking region contains several salt-responsive enhancer regions and one silencer region reflecting the complex regulation patterns exhibited by this promoter in vivo. A region localized between nucleotides -977 and -487 relative to the transcriptional start site appears to regulate the magnitude of salt-inducibility. In contrast, the Gap1 promoter contains a single region from -735 to -549 that confers salt-responsive gene expression. Alignment of these 5'-flanking regions reveals several common sequence motifs that resemble consensus binding sites for the Myb class of transcription factors. Electrophoretic gel mobility shift assays indicate that both the -877 to -679 region of Ppc1 and the -735 to -549 region of Gap1 form a DNA-protein complex unique to nuclear extracts from salt-stressed plants. The appearance of this DNA-protein complex upon salt stress suggests that it may participate in salt-induced transcriptional activation of Ppc1 and Gap1.
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