Molecular characterization of a cDNA encoding putative calcium binding protein, HvCaBP1, induced during kernel development in barley (Hordeum vulgare L.).

ABSTRACT

Seed development is known to involve complex physiological and molecular events. In order to gain information on the molecular events that occur in the grains of barley during kernel development, we conducted suppression subtractive hybridization (SSH) using grains of barley cv. Karl at 14 days after fertilization (DAF) as the tester and grains at 5 DAF as the driver. We isolated an SSH clone that showed homology with a specific calcium binding protein in rice called EFA27. Screening the cDNA library, we identified two clones as a calcium binding protein. These clones, each carrying one calcium-binding EF-hand motif, were designated HvCaBP1 (Hordeum vulgare Calcium Binding Protein 1). HvCaBP1 possesses an N-terminal region with a conserved single Ca(2+)-binding EF-hand motif and one transmembrane helix. Northern hybridization showed that the highest expression occurred in grains and that expression increased in kernels at 8 DAF. As shown in situ hybridization, the HvCaBP1 gene was highly expressed in the embryo and tissues of the endosperm near the embryo and was detected in the vascular tissues of the glume in the kernel at 8 DAF. Accumulation of HvCaBP1 mRNAs subsequently increased in vegetative tissues for up to 48 h after abscisic acid (ABA) treatment. Transcripts of HvCaBP1 mRNAs may be regulated by endogenous ABA in the grains during kernel development.