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• Class I TREHALOSE-PHOSPHATE-SYNTHASE (TPS) genes affect salinity tolerance and plant development. However, the function of class II TPS genes and their underlying mechanisms of action are unknown. • We report the identification and functional analysis of a rice class II TPS gene (OsTPS8). The ostps8 mutant was characterised by GC-MS analysis, an abscisic acid (ABA) sensitivity test and by generating transgenic lines. To identify the underlying mechanism, gene expression analyses, genetic complementation and examination of suberin deposition in the roots were conducted. • The ostps8 mutant showed salt sensitivity, ABA sensitivity and altered agronomic traits compared to the wild-type (WT), which could be rescued upon complementation. The dsRNAi line phenocopied the mutant, while the overexpression lines exhibited enhanced salt tolerance. The ostps8 mutant showed significantly reduced soluble sugars, Casparian bands and suberin deposition in the roots compared to the WT and overexpression lines. The mutant also showed downregulation of SAPKs (rice SnRK2s) and ABA-responsive genes. Furthermore, ostps8pUBI::SAPK9 rescued the salt-sensitive phenotype of ostps8. • Our results suggest that OsTPS8 may regulate suberin deposition in rice through ABA signalling. Additionally, SAPK9-mediated regulation of altered ABA-responsive genes helps to confer salinity tolerance. Overexpression of OsTPS8 was adequate to confer enhanced salinity tolerance without any yield penalty, suggesting its usefulness in rice genetic improvement.

Background Family members of sucrose non-fermenting 1-related kinase 2 (SnRK2), being plant-specific serine/threonine protein kinases, constitute the central core of abscisic acid (ABA)-dependent and ABA-independent signaling pathways, and are key regulators of abiotic stress adaptation in plants. We report here the functional characterization of SAPK9 gene, one of the 10 SnRK2s of rice, through developing gain-of-function and loss-of-function phenotypes by transgenesis. Results The gene expression profiling revealed that the abundance of single gene-derived SAPK9 transcript was significantly higher in drought-tolerant rice genotypes than the drought-sensitive ones, and its expression was comparatively greater in reproductive stage than the vegetative stage. The highest expression of SAPK9 gene in drought-tolerant Oryza rufipogon prompted us to clone and characterise the CDS of this allele in details. The SAPK9 transcript expression was found to be highest in leaf and upregulated during drought stress and ABA treatment. In silico homology modelling of SAPK9 with Arabidopsis OST1 protein showed the bilobal kinase fold structure of SAPK9, which upon bacterial expression was able to phosphorylate itself, histone III and OsbZIP23 as substrates in vitro. Transgenic overexpression (OE) of SAPK9 CDS from O. rufipogon in a drought-sensitive indica rice genotype exhibited significantly improved drought tolerance in comparison to transgenic silencing (RNAi) lines and non-transgenic (NT) plants. In contrast to RNAi and NT plants, the enhanced drought tolerance of OE lines was concurrently supported by the upgraded physiological indices with respect to water retention capacity, soluble sugar and proline content, stomatal closure, membrane stability, and cellular detoxification. Upregulated transcript expressions of six ABA-dependent stress-responsive genes and increased sensitivity to exogenous ABA of OE lines indicate that the SAPK9 is a positive regulator of ABA-mediated stress signaling pathways in rice. The yield-related traits of OE lines were augmented significantly, which resulted from the highest percentage of fertile pollens in OE lines when compared with RNAi and NT plants. Conclusion The present study establishes the functional role of SAPK9 as transactivating kinase and potential transcriptional activator in drought stress adaptation of rice plant. The SAPK9 gene has potential usefulness in transgenic breeding for improving drought tolerance and grain yield in crop plants.