Structural and functional properties of Viviparous1 genes in dormant wheat

Viviparous 1 (Vp1) of maize is known to encode a transcription factor VP1 that controls seed germination. Hexaploid wheat possesses three Vp1 homoeologues (TaVp1): TaVp-A1, TaVp-B1 and TaVp-D1. In this study, we attempted to characterize the molecular properties of TaVp1 in a highly dormant wheat cultivar, Minamino-komugi (Minamino). The seeds of Minamino showed much higher sensitivity to the inhibitory effect of ABA on germination than those of non-dormant cultivars, Sanin-1 and Tozan-18. The sequence analyses of cDNAs also revealed that some of TaVp-A1 transcripts and TaVp-D1 transcripts were spliced incorrectly, presumably resulting in production of truncated or deleted proteins. Most TaVp-B1 transcripts were spliced correctly, but some had an additional 3-bp (AAG) insertion in the B3 domain, which may not affect their function. RT-PCR analyses showed that TaVp1 was highly expressed in Minamino embryos in maturing seeds but much less in roots and leaves of seedlings. The level of TaVp1 mRNA was high when the embryos were treated with ABA but markedly decreased in water-imbibed mature embryos whose dormancy had been broken. Expression analyses of the individual homoeologues showed that the level of TaVp-A1 transcripts was highest in embryos of DAP 20 but much lower in the matured embryos. TaVp-B1 was highly expressed in developing and maturing seed embryos, while TaVp-D1 mRNA existed at lower levels in developing embryos but increased as the seeds were matured. These results suggest that the majority of TaVp1, especially TaVp-B1, are properly spliced and may function as a transcription factor playing an important role on dormancy in Minamino. By employing an efficient transient expression system using diploid wheat seeds, we confirmed the dual function of TaVP-B1: the activation of Em expression and the repression of alpha-amylase expression.