Cycloheximide induces a subset of low temperature-inducible genes in maize

To investigate further the signal transduction pathway that mediates the cold-stress response in maize, we isolated a low temperature-inducible cDNA clone (ZmCDPK1) that encodes a calcium-dependent protein kinase. Time-course experiments revealed that the low-temperature induction of ZmCDPK1 precedes that of mlip15, another cold-inducible gene that codes for a DNA-binding protein of the basic region/leucine zipper (bZIP) type, indicating that ZmCDPK1 might be located upstream of mlip15 in the cold-stress signal transduction pathway. We observed that the steady-state mRNA level of mlip15 drastically increased after cycloheximide treatment. In addition to mlip15, cycloheximide elevates the transcript levels of two other low temperature-induced genes, ZmCDPK1, and Adh1, which encodes alcohol dehydrogenase 1. In contrast, the chalcone synthase gene was only inducible by low temperature. The accumulation of the mlip15 transcript at low temperatures and in response to cycloheximide was significantly reduced by pretreatment with a calcium chelator, suggesting that calcium is involved in both cases of mlip15 induction. The signal transduction pathways triggered by low temperature and cycloheximide are discussed in relation to these observations.