okadaic-acid and lanthanum-chloride

We report unique desiccation-associated ABA signaling transduction through which the Rop (Rho GTPase of plants) and its target LLP12-2 are regulated during the stage of pollen maturation and tube growth. Overexpression of LLP12-2 drastically inhibited pollen germination and tube growth. Studies on the germination inhibitors, Ca (2+) influx blocking agents LaCl 3 and EGTA and an actin-depolymerizing drug, latrunculin B (LatB), revealed that the LLP12-2-induced inhibition of germination and tube growth is significantly suppressed by LaCl 3 and EGTA in the LLP12-2-overexpressing pollen but not by LatB. These results suggested that LLP12-2 is associated with Ca (2+) influx in the cytoplasm and may be not with actin assembly. With the addition of LaCl 3 and EGTA, LLP12-2-overexpressing pollen increased germination and tube growth compared with the one without addition, whereas pollen expressing GFP decreased germination and tube growth. Thus, an optimum level of [Ca (2+) ]cyt influx is crucial for normal germination and tube growth. Studies on the inhibitors, staurosporine and okadaic acid in the LLP12-2-overexpressing pollen, showed no appreciable increase in germination when compared with the one without addition, suggesting that staurosporine-sensitive protein kinases and dephosphorylation of phosphoproteins may be not involved in the LLP12-2 mediated germination. However, the LLP12-2-induced inhibition of tube length was slightly but significantly suppressed by staurosporine, suggesting that staurosporine-sensitive protein kinases involve in the LLP12-2-induced inhibition of tube growth.

Topics: Abscisic Acid; Arabidopsis; Calcium; cdc42 GTP-Binding Protein; Egtazic Acid; Gene Expression Regulation, Plant; Lanthanum; Okadaic Acid; Phosphoproteins; Pollen; Signal Transduction; Water

In order to determine if components of the signal transduction pathway are involved in starch metabolism during the gravitropic response, the effects of inhibitors of phosphoprotein phosphatases and protein kinases (OA), and calcium channel blockers (LaCl3), on gravitropic bending and starch levels in gravisensitive node/pulvini of oat shoots were examined. Among the compounds tested, okadaic acid (OA) and lanthanum chloride (LaCl3) showed the strongest inhibitory effects on the negative gravitropic curvature response in oat shoot node/pulvini. At the same time, they caused a rapid loss of starch in graviresponding pulvini based on a quantitative analysis of starch levels in the bending tissues over 48 h periods. These two compounds act initially to block the net increase in starch content that occurs during the early stages (0-9 h) in graviresponding oat shoot pulvini. As a result, starch levels drop precipitously in shoots treated with OA and LaCl3, starting at time zero of gravistimulation by reorientation. These findings suggest that protein dephosphorylation and calcium play a role in starch metabolism in oat shoot pulvini in response to a gravistimulation signal. They also indicate that the amount of starch present in the chloroplast gravisensors in oat shoot pulvini may determine the rate of upward bending in graviresponding pulvini.

Topics: Avena; Calcium Channel Blockers; Chloroplasts; Enzyme Inhibitors; Gravitropism; Lanthanum; Okadaic Acid; Phosphoprotein Phosphatases; Phosphorylation; Plant Proteins; Plant Shoots; Signal Transduction; Starch; Staurosporine