Transcriptional regulation of the V-ATPase subunit c and V-PPase isoforms in Cucumis sativus under heavy metal stress.

Two electrogenic proton pumps, vacuolar H(+) transporting ATPase (V-ATPase, EC 3.6.3.14) and vacuolar H(+) transporting inorganic pyrophosphatase (V-PPase, EC 3.6.1.1), co-exist in the vacuolar membrane of plant cells. In this work, all CsVHA and CsVHP genes encoding V-ATPase and V-PPase, respectively, were identified in the cucumber genome. Among them, three CsVHA-c genes for V-ATPase subunit c and two CsVHP1 genes for type I V-PPase were analyzed in detail. Individual isogenes were differentially regulated in plant tissues and during plant development as well as under changing environmental conditions. CsVHA-c1 and CsVHA-c2 showed similar tissue-specific expression patterns with the highest levels in stamens and old leaves. CsVHP1;1 was predominantly expressed in roots and female flowers. In contrast, both CsVHA-c3 and CsVHP1;2 remained in a rather constant ratio in all examined cucumber organs. Under heavy metal stress, the transcript amount of CsVHA-c1 and CsVHP1;1 showed a pronounced stress-dependent increase after copper and nickel treatment. CsVHA-c3 was upregulated by nickel only whereas CsVHA-c2 was induced by all metals with the most visible effect of copper. Additionally, CsVHP1;2 showed a tendency to be upregulated by copper and zinc. We propose that CsVHA-c1, CsVHA-c2 and CsVHP1;1 are essential elements of mechanisms involved in adaptation of cucumber plants to copper toxicity.