SLENDER RICE1 and Oryza sativa INDETERMINATE DOMAIN2 Regulating OsmiR396 Are Involved in Stem Elongation.

GAs play key roles in controlling cell proliferation through the GIBBERELLIN INSENSITIVE DWARF1/DELLA-mediated pathway. However, how DELLA proteins affect downstream pathways is not well understood. Therefore, discovering the signaling events downstream of DELLAs is key to better understanding the roles of GAs in plant development. Here, we discovered that miR396 is regulated by SLENDER RICE1 (SLR1) in controlling cell proliferation. The positive response of rice (Oryza sativa) GROWTH-REGULATING FACTORs (OsGRFs) to GAs was found to be caused by a negative response of miR396 to GAs. miR396 acts downstream of SLR1 and upstream of GA-induced cell-cycle genes. Rice INDETERMINATE DOMAIN2 (OsIDD2) directly binds the promoter of OsmiR396a and can interact with SLR1 in vivo and in vitro. Rice lines overexpressing miR396a (miR396OE) or OsIDD2 (OsIDD2OE) displayed dwarfism resulting from higher abundance of miR396 RNA. However, the stem elongation of OsIDD2OE plants could be significantly stimulated by applying exogenous GA3, while that of miR396OE plants could not. Rice with OsIDD2 knocked down by RNA interference showed a slr1-like phenotype, in which the expression of miR396 was inhibited while its targets were enhanced. The protein levels of OsIDD2 were unaffected by GA in wild-type and OsIDD2OE plants, implying that OsIDD2 promotes the expression of miR396 and likely requires the coactivator of SLR1. Taken together, these results provided a close link between SLR1/OsIDD2 and GRFs via a negative regulator, miR396, and thus highlighted a molecular mechanism of GA-mediated cell proliferation in rice.

Coordination between GROWTH-REGULATING FACTOR1 and GRF-INTERACTING FACTOR1 plays a key role in regulating leaf growth in rice.

BACKGROUND: The interactions between Growth-regulating factors (GRFs) and GRF-Interacting Factors (GIFs) have been well demonstrated but it remains unclear whether different combinations of GRF and GIF play distinctive roles in the pathway downstream of the complex. RESULTS: Here we showed that OsGRF1 and OsGIF1 synergistically regulate leaf growth in rice. The expression of OsGIF1 emerged in all tissues with much higher level while that of OsGRF1 appeared preferentially only in the stem tips containing shoot apical meristem (SAM) and younger leaves containing leaf primordium. Overexpression of an OsmiR396-resistant version of mOsGRF1 resulted in expanded leaves due to increased cell proliferation while knockdown of OsGRF1 displayed an opposite phenotype. Overexpression of OsGIF1 did not exhibit new phenotype while knockdown lines displayed pleiotropic growth defects including shrunken leaves. The crossed lines of mOsGRF1 overexpression and OsGIF1 knockdown still exhibited shrunk leaves, indicating that OsGIF1 is indispensable in leaf growth regulated by OsGRF1. The expression of OsGRF1 could be upregulated by gibberellins (GAs) and downregulated by various stresses while that of OsGIF1 could not. CONCLUSION: Our results suggest that OsGIF1 is in an excessive expression in various tissues and play roles in various aspects of growth while OsGRF1 may specifically involve in leaf growth through titrating OsGIF1. Both internal and external conditions impacting leaf growth are likely via way of regulating the expression of OsGRF1.

MiR393 and miR390 synergistically regulate lateral root growth in rice under different conditions.

BACKGROUND: Plants have evolved excellent ability of flexibly regulating the growth of organs to adapt to changing environment, for example, the modulation of lateral root development in response to environmental stresses. Despite of fundamental discovery that some microRNAs are involved in this process, the molecular mechanisms of how these microRNAs work together are still largely unknown. RESULTS: Here we show that miR390 induced by auxin promotes lateral root growth in rice. However, this promotion can be suppressed by miR393, which is induced by various stresses and ABA (Abscisic Acid). Results that miR393 responded to ABA stronger and earlier than other stresses implied that ABA likely is authentic factor for inducing miR393. The transgenic lines respectively over-expressing miR393 and OsTAS3a (Oryza sativa Trans-Acting Short RNA precursor 3a) displayed opposite phenotypes in lateral root growth. MiR390 was found to be dominantly expressed at lateral root primordia and roots tips while miR393 mainly expressed in the base part of roots at very low level. When miR393 was up-regulated by various stresses, miR390 expression level fell down. However, the risen expression level of miR390 induced by auxin didn't affect the expression of miR393 and its target OsTIR1 (Transport Inhibitor Response 1). Together with analysis of the two transgenic lines, we provide a model of how the growth of lateral roots in rice is regulated distinctively by the 2 microRNAs. CONCLUSION: We propose that miR390 induced by auxin triggers the lateral root growth under normal growth conditions, meanwhile miR393 just lurks as a potentially regulative role; Once plants suffer from stresses, miR393 will be induced to negatively regulate miR390-mediated growth of lateral roots in rice.

miR398 regulation in rice of the responses to abiotic and biotic stresses depends on CSD1 and CSD2 expression.

MiR398 targets two Cu or Zn superoxide dismutases (CSD1 and CSD2) in Arabidopsis thaliana (L.) Heynh. Here we provide evidence that rice (Oryza sativa L.) miR398 mediates responses to abiotic and biotic stresses through regulating the expression of its target genes, Os-CSD1 and Os-CSD2. Rice plants were exposed to various stresses, including high Cu2+, high salinity, high light, methyl viologen, water stress, pathogens and ethylene, and the molecular response was investigated. Rice plants overexpressing Os-miR398 and the miR398-resistant form of Os-CSD2 were also exposed to these stresses. Both abiotic and biotic stresses significantly inhibited Os-miR398 expression and thus stimulated the expression of Os-CSD1 and Os-CSD2. The plant hormone ethylene produced an especially marked response. Transgenic rice lines that overexpressed Os-miR398 had a lower expression of CSD1 and -2 and were more sensitive to environmental stress. Conversely, transgenic rice lines which overexpressed the miR398-resistant form of Os-CSD2 showed more tolerance to high salinity and water stress than non-transgenic rice. We conclude that Os-miR398 regulates the responses of rice to a wide range of environmental stresses and to ethylene, and exerts its role through mediating CSDs expression and cellular ROS levels.