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Free keywords:
abscisic acid; coiled-coil; Craterostigma; desiccation tolerance; green fluorescent protein-fusion; plastid-targeted protein
Abstract:
In the desiccation-tolerant resurrection plant Craterostigma plantagineum Hochst. the chloroplasts undergo major ultrastructural changes during dehydration, which are reversible upon rehydration. Such alterations argue the need for efficient protective/stabilising mechanisms to exist. Here we describe a novel gene family that is rapidly and transiently expressed in response to both dehydration and exogenously applied abscisic acid, mostly in the chloroplast-rich palisade layer on the adaxial side of the leaf. Analysis of the putative coding region suggests that the resulting protein is plastid- targeted. This was confirmed using a chimeric green fluorescent protein (GFP) reporter construct in transgenic tobacco plants - hence the gene family is termed Plastid Targeted Protein (CpPTP). Fluorescence microscopy also revealed that CpPTP was localised in structures similar to proplastid nucleoids in transgenic tobacco (Nicotiana tabacum L.) BY-2 cells. The ability of CpPTP to interact with DNA was demonstrated through a DNaseI protection assay. A structure-prediction programme suggests that the mature CpPTP is composed almost entirely of a pattern of hydrophobic and hydrophilic residues that form heptad repeats. which are the hallmarks of a coiled-coil domain. Given the localisation and DNA-binding property of the protein, we propose that CpPTP plays a role during the early stages of dehydration-induced chloroplast remodelling.
