Cloning and expression of two plasma membrane aquaporins expressed during the ripening of grape berry

Abstract
The ripening of grape (Vitis vinifera L.) berry is accompanied by dramatic accumulation of sugars and water. Two full-length clones and several partial clones encoding plasma membrane aquaporins (PIP) were cloned from grape berries collected at the beginning of ripening. Based on their sequences, on a phylogenetic analysis and on functional properties, both clones, called VvPIP1a and VvPIP1b were assigned to the PIP1 subfamily. RNA gel blot studies with berries at various stages of development indicated that VvPIP expression was highest at stages following veraison. Injection of Xenopus oocytes with VvPIP1a cRNA induced a moderate increase of water permeability and a large increase in glycerol permeability, whereas injection with VvPIP1b cRNA did not affect these permeabilities. Injection of VvPIP1a cRNA, but not VvPIP1b cRNA, inhibited urea uptake by the oocyte, and this inhibition was sensitive to HgCl2. The data are discussed in relation with the potential role of aquaporins in fruit physiology.