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Transcriptome analyses of Arabidopsis lines overexpressing distinct PLC genes to explore their contribution to drought tolerance

ABSTRACT: Overexpression (OE) of phospholipase C (PLC) promotes drought stress tolerance in various plant species. How PLC contributes to this phenotype is unknown. To investigate this, transcriptomic analyses of drought-stressed Arabidopsis plants were performed in wild type (WT) and four distinct PLC-OE backgrounds. To be able to measure the earliest responses in both root- and shoot tissues, a vermiculite-based assay was developed. Subsequent transcriptomics analyses revealed that WT plants responded relatively strongly to the drought, whereas all PLC-OEs lines responded to a lesser extent. Changes in various pathways were observed that were differentially regulated in WT. These include, i) several phospholipid metabolism genes, which may counteract the overexpression of PLC activity, ii) genes involved in the circadian rhythm, shifting to a more night-like expression pattern for the PLC-OE lines, iii) downregulation of several light reaction-related genes, and iv) upregulation of genes involved in the brassinosteroid signalling pathway. In the PLC-OE lines, a subset of genes involved in auxin signalling was found to be differently regulated under drought. Similarly, genes upstream of H2O2 production were found upregulated, while ROS-induced genes were downregulated. Though unable to pin point the primary cause of the drought tolerant phenotype in PLC-OE plants, several novel connections were identified, which are highlighted and discussed in more detail.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE270544 | GEO | 2024/07/01

REPOSITORIES: GEO

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Project description:Drought is one of the major abiotic stresses threatening rice (Oryza sativa) production worldwide. Drought resistance is controlled by multiple genes, and therefore, a multi-gene genetic engineering strategy is theoretically useful for improving drought resistance. However, the experimental evidence for such a strategy is still lacking. In this study, a few drought-responsive genes from rice were assembled by a multiple-round site-specific assembly (MISSA) system, and the constructs were introduced into the rice cultivar KY131 via Agrobacterium-mediated transformation. The transgenic lines of the multi-gene and corresponding single-gene constructs were pre-evaluated for drought resistance. We found that the co-overexpression of two genes, encoding a constitutively active form of a bZIP transcription factor (OsbZIP46CA1) and a protein kinase (SAPK6) involved in the abscisic acid (ABA) signaling pathway, showed significantly enhanced drought resistance compared with the single-gene transgenic lines and the negative transgenic plants. Single-copy lines of this bi-gene combination (named XL22) and the corresponding single-gene lines were further evaluated for drought resistance in the field using agronomical traits. The results showed that XL22 exhibited greater yield, biomass, spikelet number, and grain number under moderate drought stress conditions. The seedling survival rate of XL22 and the single-gene overexpressors after drought stress treatment also supported the drought resistance results. Furthermore, expression profiling by RNA-Seq revealed that many genes involved in the stress response were specifically up-regulated in the drought-treated XL22 lines and some of the stress-related genes activated in CA1-OE and SAPK6-OE were distinct, which could partially explain the different performances of these lines with respect to drought resistance. In addition, the XL22 seedlings showed improved tolerance to heat and cold stresses. Our results demonstrate that the multi-gene assembly in an appropriate combination may be a promising approach in the genetic improvement of drought resistance.

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Transcriptome analyses of Arabidopsis lines overexpressing distinct PLC genes to explore their contribution to drought tolerance

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