Plant organ development is important for adaptation to a changing environment. Genetic and physiological studies have revealed that plant hormones play key roles in lateral root formation. In this study, we show that MIZU-KUSSEI1 (MIZ1), which was identified originally as a regulator of hydrotropism, functions as a novel regulator of hormonally mediated lateral root development. Overexpression of MIZ1 (MIZ1OE) in roots resulted in a reduced number of lateral roots being formed; however, this defect could be recovered with the application of auxin. Indole-3-acetic acid quantification analyses showed that free indole-3-acetic acid levels decreased in MIZ1OE roots, which indicates that alteration of auxin level is critical for the inhibition of lateral root formation in MIZ1OE plants. In addition, MIZ1 negatively regulates cytokinin sensitivity on root development. Application of cytokinin strongly induced the localization of MIZ1-green fluorescent protein to lateral root primordia, which suggests that the inhibition of lateral root development by MIZ1 occurs downstream of cytokinin signaling. Surprisingly, miz2, a weak allele of gnom, suppressed developmental defects in MIZ1OE plants. Taken together, these results suggest that MIZ1 plays a role in lateral root development by maintaining auxin levels and that its function requires GNOM activity. These data provide a molecular framework for auxin-dependent organ development in Arabidopsis (Arabidopsis thaliana).

Plant roots undergo tropic growth in response to environmental cues, and each tropic response is affected by several environmental stimuli. Even its importance, molecular regulation of hydrotropism has not been largely uncovered. Tropic responses including hydrotropism were impacted by other environmental signal. We found that hydrotropism was reduced in dark-grown seedling. Moreover, we found that the expression of MIZ1, an essential gene for hydrotropism, was regulated by light signal. From our genetic analysis, phytochrome A (phyA)-, phyB-and HY5-mediated blue-light signalling play curial roles in lightmediated induction of MIZ1 and hydrotropism. In addition, we found that abscisic acid (ABA) also induced MIZ1 expression. ABA treatment could recover weak hydrotropism and MIZ1 expression level of hy5, and ABA synthesis inhibitor, abamineSG, further reduced hydrotropic curvature of hy5. In contrast, ABA treatment did not affect ahydrotropic phenotype of miz1. These results suggest that ABA signalling regulates MIZ1 expression independently from light signalling. Our results demonstrate that environmental signals, such as light and stresses mediated by ABA signalling, are integrated into MIZ1 expression and thus regulate hydrotropism. These machineries will allow plants to acquire sufficient amounts of water.

Roots show positive hydrotropism in response to moisture gradients, which is believed to contribute to plant water acquisition. This article reviews the recent advances of the physiological and molecular genetic studies on hydrotropism in seedling roots of Arabidopsis thaliana. We identified MIZU-KUSSEI1 (MIZ1) and MIZ2, essential genes for hydrotropism in roots; the former encodes a protein of unknown function, and the latter encodes an ARF-GEF (GNOM) protein involved in vesicle trafficking. Because both mutants are defective in hydrotropism but not in gravitropism, these mutations might affect a molecular mechanism unique to hydrotropism. MIZ1 is expressed in the lateral root cap and cortex of the root proper. It is localized as a soluble protein in the cytoplasm and in association with the cytoplasmic face of endoplasmic reticulum (ER) membranes in root cells. Light and ABA independently regulate MIZ1 expression, which influences the ultimate hydrotropic response. In addition, MIZ1 overexpression results in an enhancement of hydrotropism and an inhibition of lateral root formation. This phenotype is likely related to the alteration of auxin content in roots. Specifically, the auxin level in the roots decreases in the MIZ1 overexpressor and increases in the miz1 mutant. Unlike most gnom mutants, miz2 displays normal morphology, growth, and gravitropism, with normal localization of PIN proteins. It is probable that MIZ1 plays a crucial role in hydrotropic response by regulating the endogenous level of auxin in Arabidopsis roots. Furthermore, the role of GNOM/MIZ2 in hydrotropism is distinct from that of gravitropism.

a b s t r a c tMIZ1 is encoded by a gene essential for root hydrotropism in Arabidopsis. To characterize the property of MIZ1, we used transgenic plants expressing GFP-tagged MIZ1 (MIZ1-GFP) and mutant MIZ1 (MIZ1 G235E -GFP) in a miz1-1 mutant. Although both chimeric genes were transcribed, the translational products of MIZ1 G235E -GFP did not accumulate in roots. Moreover, MIZ1-GFP complemented the mutant phenotype but not MIZ1 G235E -GFP. The signal corresponding to MIZ1-GFP was detected at high levels in cortical cells and lateral root cap cells and accumulated in compartments in cortical cells. MIZ1-GFP was fractionated into a soluble protein fraction and an endoplasmic reticulum (ER) membrane fraction, where it was bound to the surface of the ER membrane at the cytosolic side.