Regulated proteolysis by the ubiquitin-26S proteasome system challenges transcription and phosphorylation in magnitude and is one of the most important regulatory mechanisms in plants. This article describes the characterization of a rice () auxin-responsive Kelch-domain-containing F-box protein, OsFBK1, found to be a component of an SCF E3 ligase by interaction studies in yeast. Rice transgenics of displayed variations in anther and root secondary cell wall content; it could be corroborated by electron/confocal microscopy and lignification studies, with no apparent changes in auxin content/signaling pathway. The presence of U-shaped secondary wall thickenings (or lignin) in the anthers were remarkably less pronounced in plants overexpressing as compared to wild-type and knockdown transgenics. The roots of the transgenics also displayed differential accumulation of lignin. Yeast two-hybrid anther library screening identified an OsCCR that is a homolog of the well-studied Arabidopsis () IRX4; OsFBK1-OsCCR interaction was confirmed by fluorescence and immunoprecipitation studies. Degradation of OsCCR mediated by SCF and the 26S proteasome pathway was validated by cell-free experiments in the absence of auxin, indicating that the phenotype observed is due to the direct interaction between OsFBK1 and OsCCR. Interestingly, the knockdown transgenics also displayed a decrease in root and anther lignin depositions, suggesting that OsFBK1 plays a role in the development of rice anthers and roots by regulating the cellular levels of a key enzyme controlling lignification.

Traditional cultivars of rice in India exhibit tolerance to drought stress due to their inherent genetic variations. Here we present comparative physiological and transcriptome analyses of two contrasting cultivars, drought tolerant Dhagaddeshi (DD) and susceptible IR20. Microarray analysis revealed several differentially expressed genes (DEGs) exclusively in DD as compared to IR20 seedlings exposed to 3 h drought stress. Physiologically, DD seedlings showed higher cell membrane stability and differential ABA accumulation in response to dehydration, coupled with rapid changes in gene expression. Detailed analyses of metabolic pathways enriched in expression data suggest interplay of ABA dependent along with secondary and redox metabolic networks that activate osmotic and detoxification signalling in DD. By co-localization of DEGs with QTLs from databases or published literature for physiological traits of DD and IR20, candidate genes were identified including those underlying major QTL qDTY1.1 in DD. Further, we identified previously uncharacterized genes from both DD and IR20 under drought conditions including OsWRKY51, OsVP1 and confirmed their expression by qPCR in multiple rice cultivars. OsFBK1 was also functionally validated in susceptible PB1 rice cultivar and Arabidopsis for providing drought tolerance. Some of the DEGs mapped to the known QTLs could thus, be of potential significance for marker-assisted breeding.

Auxin is among the key growth regulators that play diverse roles in virtually all aspects of plant growth and development. Intensive investigations during the past two decades have helped in elucidation of auxin perception and signal transduction mechanisms operative in plants. In addition to its primary role in regulating plant development, several studies in recent years have provided unfl inching evidence for the involvement of auxin in abiotic stress responses. Functional genomics studies and genome-wide expression analysis have revealed altered expression of auxin-responsive genes, such as Aux/IAA , GH3 , SAURs , and ARFs , under abiotic stress conditions . Variations in endogenous levels of auxin at global and local levels under various abiotic stress conditions have been associated with phenotypic changes and provided intriguing evidences regarding its role in response to environmental changes. Modulation of reactive oxygen species (ROS) levels in response to exogenous auxin as well as to drought, salinity, and ABA have indicated towards a complex relationship network between auxin, ROS, and abiotic stresses in plants. The advent of recent functional genomics technologies has led to identifi cation of several candidate genes that may modulate crosstalk between auxin and abiotic stresses. This chapter discusses auxin homeostasis, signal transduction mechanisms, and how these processes are modulated under abiotic stresses, thus emphasizing on the emerging roles of auxin as a key integrator of abiotic stress pathways and plant development.

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We had previously shown the rice F-box, OsFBK1, plays a role in anther development by mediating the turnover of OsCCR14, a cinnamoyl CoA-reductase regulating lignification. Another substrate identified in the same Y2H library screening was OsATL53, a member of the ATL family of RING-H2 proteins that is primarily localized to the cytoplasm. We found OsATL53 to be a component and substrate of SCFOsFBK1 by immunoprecipitation and cell-free studies. Incidentally, OsATL53 was found to interact with OsCCR14 in the cytoplasm and form a stable complex in cell-free experiments and bimolecular fluorescence complementation assays. Biochemically, OsATL53 was found to influence the enzymatic activity of OsCCR14 by decreasing its efficiency. Degradation studies have shown OsFBK1 mediates turnover of OsCCR14 in the nucleus, while OsATL53 is degraded in both cytoplasm and nucleus. The degradation of ATLs by F-box proteins has not been reported before. In presence of jasmonic acid (JA), which plays a role in anther dehiscence, OsATL53 has been found to gather around the nucleus, and this property enables the translocation of the OsATL53-OsCCR14 complex from a cytoplasmic localization to accumulate around the nuclear periphery. FLIM analyses revealed OsCCR14-OsATL53 complex undergoing conformational changes in presence of JA and this triggers OsFBK1 to mediate the targeted degradation of OsATL53 in the cytoplasm, thereby dissociating the cytoplasmic OsCCR14-OsATL53 complex and enabling OsCCR14 to enter the nucleus and eventually get degraded by SCFOsFBK1 E3 ligase. We have thus studied the signalling mechanism of a variant JA-induced E3 ligase-mediated substrate turnover in plants at the molecular level.