Spatiotemporal activities of Douglas-fir BiP Pro1 promoter in transgenic potato.

Abstract

The PmBiPPro1 promoter of the luminal binding protein (BiP) from Douglas-fir is fully functional in transgenic potato, responsive to wounding, and has high transcriptional activity in tubers. A predefined pattern and level of transgene expression targeted to specific tissues or organs and at a particular developmental stage is a pre-requisite for the successful development of plants with desired traits. Here, we evaluated the transcriptional activity of the PmBiPPro1 promoter of the luminal binding protein (BiP) from Douglas-fir, by expressing reporter β-D-glucuronidase (GUS) gene constructs containing three different PmBiPPro1 promoter versions (2258bp, 1259bp, and 278bp) in transgenic potato. In conifers, this promoter regulates the endoplasmic reticulum (ER) molecular chaperon of the HSP70 stress-related protein family and is essential for proper functioning of the ER. Stable expression analysis demonstrated that two of three PmBiPPro1 promoter versions (PmBiPPro1-1 and PmBiPPro1-3) were fully functional in the heterologous host, exhibited high transcriptional activities in the leaves of unstressed potatoes, and were responsive to wounding. Deletion analysis showed that the positive cis-active regulatory elements necessary for higher level expression resided within the - 1243 to - 261 region, whereas negative cis-active elements encompassed nucleotides - 2242 to - 1243. Histochemical staining revealed high level of GUS activities in tissues associated with a high rate of cell division and secretory activities. Most importantly, the PmBiPPro1 promoters, especially the full-length version, had activity in microtubers at a level that was much higher than in any other potato organ or tissue. The - 2242 to - 1243bp region likely contains important cis element(s) that interact with tuber-specific transcription factors required for promoter activation in the storage organs. The organ-specific activity of the PmBiPPro1 promoters may be useful for targeted expression of heterologous molecules in potato tubers.