Expression of a cauliflower tonoplast aquaporin tagged with GFP in tobacco suspension cells correlates with an increase in cell size.

Abstract

In plants, vacuoles are essential organelles that undergo dynamic volume changes during cell growth due to rapid and high flow of water through tonoplast water-carrying channels composed of integral proteins (tonoplast aquaporins). The tonoplast BobTIP26-1 from cauliflower has previously been shown to be an efficient active aquaporin in Xenopus leavis oocytes. In this study we used tobacco (Nicotiana tabacum cv. Wisconsin 38) suspension cells to examine the effect of BobTIP26-1 expression. In order to follow the intracellular localisation of the protein in real time, the gfp sequence was fused downstream to the BobTIP26-1 coding region. The fusion protein BobTIP26-1::GFP is less active than BobTIP26-1 by itself when expressed in Xenopus oocytes. Nevertheless, this fusion protein is well targeted to the tonoplast of the plant suspension cell when expressed via Agrobacterium co-cultivation. A complex tonoplast labelling is shown when young vacuolated cells are observed. The expression of the fusion protein does not affect the growth rate of the cells but increases their volume. We postulate that the increase in cell volume is triggered by the fusion protein allowing vacuolar volume increase.