Cold-shock regulation of the arabidopsis TCH genes and the effects of modulating intracellular calcium levels

The Arabidopsis TCH genes, which encode calmodulin-related proteins and a xyloglucan endotransglycosylase, are shown to be up-regulated in expression following cold shock. We investigated a possible role of fluctuations in intracellular calcium ion concentrations ([Ca2+]) in the cold-shock-induced TCH gene expression. Transgenic plants harboring the apoaequorin gene were generated to monitor [Ca2+] and to test the necessity of cold-induced [Ca2+] increases for TCH expression. Cold-shock-induced [Ca2+] increases can be blocked by La3+ and Gd3+, putative plasma membrane Ca2+ channel blockers, and 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, an extracellular Ca2+ chelator. Cold-shock-induced expression of the TCH genes is inhibited by levels of La3+, Gd3+, and 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-acid, that have been shown to block [Ca2+] increases. These data support the hypotheses that (a) intracellular [Ca2+] increases following cold shock require extracellular Ca2+ and may derive from a Ca2+ influx mediated by plasmalemma Ca2+ channels, and (b) cold up-regulation of expression of at least a subset of the TCH genes requires an intracellular [Ca2+] increase. The inhibitors are also shown to have stimulus-independent effects on gene expression, providing strong evidence that these commonly used chemicals have more complex effects than generally reported.