Detection of Sugar-Regulated Gene Expression and Signaling in Suspension-Cultured Rice Cells

摘要

To better understand the mechanism of sugar signaling in rice cell, the suspension-cultured rice cells were transferred from sucrose-containing (+S) to sucrose-free (-S) of MS culture medium, we found that ribosomal RNAs (rRNAs) were degraded progressively. This suggests that carbon, nitrogen, and phosphate were recycled in this process and the reduction in cellular rRNAs might lead to decreased translation to save energy in response to sugar starvation. Differential screening revealed that two groups of genes, sugar-starvation-repressed (SSR) and sugar-starvation-activated (SSA) genes, were regulated by sugar in an opposing manner. Northern-blot analysis showed that two major hybridization signals of 0.8 and 1.9 kb were induced strongly under sugar starvation. The two populations of genes corresponded with homologs of α-amylases (1.9 kb) and the glycine-rich proteins (GRPs) gene family (0.8 kb), and all were SSA genes. Expression of GRP genes was strongly induced in sugar-starved cells, which suggests that GRPs may help to protect cells against nutritional stress. Treatment of +S and -S cells with the protein kinase (PK) inhibitor staurosporine (St) and the serine/theronine phosphoprotein phosphatases 1 (PP1) and 2A (PP2A) inhibitor okadaic acid (OA) revealed that PP1 and PP2A (PPs) might be involved in increasing SSR gene expression in +S cells, and that activation of the majority of the SSA genes in -S cells might be due to PKs activity. These results suggested that PKs and PPs might be involved in the sugar regulation of SSR and SSA gene expression. An in-gel PK activity assay demonstrated that the activity of two classes of PKs (50 and 66 kDa) may be induced rapidly after transfer of +S cells to -S medium. Following transfer of -S cells to +S medium, a novel class of 38 kDa PK was induced rapidly and showed high activity. The 38 kDa PK might play a role in sugar sensing, and the 50 and 66 kDa PKs might play roles in signal sensing under sugar starvation in rice cells. These results provide valuable information on three classes of protein kinases that might play key roles in sugar sensing and signaling in rice.