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      Variations in OsSPL10 confer drought tolerance by directly regulating OsNAC2 expression and ROS production in rice

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          ABSTRACT

          Drought is a major factor restricting the production of rice ( Oryza sativa L.). The identification of natural variants for drought stress‐related genes is an important step toward developing genetically improved rice varieties. Here, we characterized a member of the SQUAMOSA PROMOTER BINDING PROTEIN‐LIKE (SPL) family, OsSPL10, as a transcription factor involved in the regulation of drought tolerance in rice. OsSPL10 appears to play a vital role in drought tolerance by controlling reactive oxygen species (ROS) production and stomatal movements. Haplotype and allele frequency analyses of OsSPL10 indicated that most upland rice and improved lowland rice varieties harbor the OsSPL10 Hap1 allele, whereas the OsSPL10 Hap2 allele was mainly present in lowland and landrace rice varieties. Importantly, we demonstrated that the varieties with the OsSPL10 Hap1 allele showed low expression levels of OsSPL10 and its downstream gene, OsNAC2, which decreases the expression of OsAP37 and increases the expression of OsCOX11, thus preventing ROS accumulation and programmed cell death (PCD). Furthermore, the knockdown or knockout of OsSPL10 induced fast stomatal closure and prevented water loss, thereby improving drought tolerance in rice. Based on these observations, we propose that OsSPL10 confers drought tolerance by regulating OsNAC2 expression and that OsSPL10 Hap1 could be a valuable haplotype for the genetic improvement of drought tolerance in rice.

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          Reactive oxygen species homeostasis and signalling during drought and salinity stresses.

          Water deficit and salinity, especially under high light intensity or in combination with other stresses, disrupt photosynthesis and increase photorespiration, altering the normal homeostasis of cells and cause an increased production of reactive oxygen species (ROS). ROS play a dual role in the response of plants to abiotic stresses functioning as toxic by-products of stress metabolism, as well as important signal transduction molecules. In this review, we provide an overview of ROS homeostasis and signalling in response to drought and salt stresses and discuss the current understanding of ROS involvement in stress sensing, stress signalling and regulation of acclimation responses.
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            Reactive oxygen species (ROS) and response of antioxidants as ROS-scavengers during environmental stress in plants

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              Transient expression vectors for functional genomics, quantification of promoter activity and RNA silencing in plants

              Background We describe novel plasmid vectors for transient gene expression using Agrobacterium, infiltrated into Nicotiana benthamiana leaves. We have generated a series of pGreenII cloning vectors that are ideally suited to transient gene expression, by removing elements of conventional binary vectors necessary for stable transformation such as transformation selection genes. Results We give an example of expression of heme-thiolate P450 to demonstrate effectiveness of this system. We have also designed vectors that take advantage of a dual luciferase assay system to analyse promoter sequences or post-transcriptional regulation of gene expression. We have demonstrated their utility by co-expression of putative transcription factors and the promoter sequence of potential target genes and show how orthologous promoter sequences respond to these genes. Finally, we have constructed a vector that has allowed us to investigate design features of hairpin constructs related to their ability to initiate RNA silencing, and have used these tools to study cis-regulatory effect of intron-containing gene constructs. Conclusion In developing a series of vectors ideally suited to transient expression analysis we have provided a resource that further advances the application of this technology. These minimal vectors are ideally suited to conventional cloning methods and we have used them to demonstrate their flexibility to investigate enzyme activity, transcription regulation and post-transcriptional regulatory processes in transient assays.
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                Author and article information

                Journal
                Journal of Integrative Plant Biology
                JIPB
                Wiley
                1672-9072
                1744-7909
                April 2023
                January 17 2023
                April 2023
                : 65
                : 4
                : 918-933
                Affiliations
                [1 ] State Key Laboratory of Agrobiotechnology/Beijing Key Laboratory of Crop Genetic Improvement, College of Agronomy and Biotechnology China Agricultural University Beijing 100193 China
                [2 ] Shanghai Key Laboratory of Plant Molecular Sciences, College of Life Sciences Shanghai Normal University Shanghai 200234 China
                [3 ] Sanya Institute of China Agricultural University Sanya 572025 China
                Article
                10.1111/jipb.13414
                36401566
                1fd0c8f0-f878-4553-8120-7b28513ebcb9
                © 2023

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