SUMMARYThe plant U-box (PUB) protein functions as an E3 ligase to poly-ubiquitinate a target protein for its degradation or post-translational modification. Here, we report functional roles for OsPUB15, which encodes a cytosolic U-box protein in the class-II PUB family. Self-ubiquitination assays showed that bacterially expressed MBPOsPUB15 protein has E3 ubiquitin ligase activity. A T-DNA insertional mutation in OsPUB15 caused severe growth retardation and a seedling-lethal phenotype. Mutant seeds did not produce primary roots, and their shoot development was significantly delayed. Transgenic plants expressing the OsPUB15 antisense transcript phenocopied these mutant characters. The abnormal phenotypes were partially rescued by two antioxidants, catechin and ascorbic acid. Germinating seeds in the dark also recovered the rootless defect. Levels of H 2 O 2 and oxidized proteins were higher in the knock-out mutant compared with the wild type. OsPUB15 transcript levels were increased upon H 2 O 2 , salt and drought stresses; plants overexpressing the gene grew better than the wild type under high salinity. These results indicate that PUB15 is a regulator that reduces reactive oxygen species (ROS) stress and cell death.

Increased grain yield will be critical to meet the growing demand for food, and could be achieved by delaying crop senescence. Here, via quantitative trait locus (QTL) mapping, we uncover the genetic basis underlying distinct life cycles and senescence patterns of two rice subspecies, indica and japonica. Promoter variations in the Stay-Green (OsSGR) gene encoding the chlorophyll-degrading Mg++-dechelatase were found to trigger higher and earlier induction of OsSGR in indica, which accelerated senescence of indica rice cultivars. The indica-type promoter is present in a progenitor subspecies O. nivara and thus was acquired early during the evolution of rapid cycling trait in rice subspecies. Japonica OsSGR alleles introgressed into indica-type cultivars in Korean rice fields lead to delayed senescence, with increased grain yield and enhanced photosynthetic competence. Taken together, these data establish that naturally occurring OsSGR promoter and related lifespan variations can be exploited in breeding programs to augment rice yield.

A genome-wide survey of Catharanthus roseus receptor-like kinase1-like kinases (CrRLK1Ls) in rice revealed that the pattern of expression by some CrRLK1Ls is controlled by drought or circadian rhythms. This is probably accomplished through the functioning of Gigantea ( OsGI ). Such findings provide a novel angle for using CrRLK1Ls to study the drought-stress response and circadian regulation. The 17 CrRLK1L members of a novel RLK family have been identified in Arabidopsis. Each carries a putative extracellular carbohydrate-binding malectin-like domain. However, their roles in rice, a widely consumed staple food, are not well understood. To investigate the functions of CrRLK1Ls in rice, we utilized phylogenomics data obtained through anatomical and diurnal meta-expression analyses. This information was integrated with a large set of public microarray data within the context of the rice CrRLK1L family phylogenic tree. Chromosomal locations indicated that 3 of 16 genes were tandem-duplicated, suggesting possible functional redundancy within this family. However, integrated diurnal expression showed functional divergence between two of three genes, i.e., peak expression was detected during the day for OsCrRLK1L2, but during the night for OsCrRLK1L3. We found it interesting that OsCrRLK1L2 expression was repressed in osgigantea (osgi) mutants, which suggests that it could function downstream of OsGI. Network analysis associated with OsCrRLK1L2 and OsGI suggested a novel circadian regulation mechanism mediated by OsGI. In addition, two of five OsCrRLK1Ls preferentially expressed in the roots were stimulated by drought, suggesting a potential role for this family in water-use efficiency. This preliminary identification of CrRLK1Ls and study of their expression in rice will facilitate further functional classifications and applications in plant production.

Root hairs are important for absorption of nutrients and water from the rhizosphere. The Root Hair Defective-Six Like (RSL) Class II family of transcription factors is expressed preferentially in root hairs and has a conserved role in root hair development in land plants. We functionally characterized the seven members of the RSL Class II subfamily in the rice (Oryza sativa) genome. In root hairs, six of these genes were preferentially expressed and four were strongly expressed. Phenotypic analysis of each mutant revealed that Os07g39940 plays a major role in root hair formation, based on observations of a short root hair phenotype in those mutants. Overexpression (OX) for each of four family members in rice resulted in an increase in the density and length of root hairs. These four members contain a transcription activation domain and are targeted to the nucleus. They interact with rice Root Hairless1 (OsRHL1), a key regulator of root hair development. When heterologously expressed in epidermal cells of Nicotiana benthamiana leaves, OsRHL1 was predominantly localized to the cytoplasm. When coexpressed with each of the four RSL Class II members, however, OsRLH1 was translocated to the nucleus. Transcriptome analysis using Os07g39940-OX plants revealed that 86 genes, including Class III peroxidases, were highly up-regulated. Furthermore, reactive oxygen species levels in the root hairs were increased in Os07g39940-OX plants but were drastically reduced in the os07g39940 and rhl1 mutants. Our results demonstrate that RSL Class II members function as essential regulators of root hair development in rice.