The transcript levels of more than 500 genes were more than 2.5-fold reduced in 35S:ARR1-SRDX transgenic seedlings, suggesting a broad function of B-type ARRs. The rapid induction of a large part of the cytokinin response genes was dampened. Our results support a model in which cytokinin regulates a wide array of downstream responses through the action of a multistep phosphorelay that culminates in transcriptional regulation by ARR1, ARR10, and ARR12. Several bioassays showed that 35S:ARR1-SRDX plants have an increased resistance toward cytokinin. Characterization of double mutants revealed differing contributions of the type B ARRs to mutant phenotypes. To get around this problem, Shikata and colleagues (Shikata et al., 2009) fused SPL10 to the SRDX transcription repression domain, and expressed this. We then generated additional repressive trans-elements by fusing the SRDX domain (-SRDX for C-terminal fusions and -SRDXn for N-terminal fusions) to the other orthogonal TFs we tested (Fig. Microarray analysis revealed that expression of the majority of cytokinin-regulated genes requires the function of ARR1, ARR10, and ARR12. In recent years, this technology has been widely used for analyzing plant functional genes of transcription factors. The triple mutant exhibited reduced stature due to decreased cell division in the shoot, enhanced seed size, increased sensitivity to light, altered chlorophyll and anthocyanin concentrations, and an aborted primary root with protoxylem but no metaxylem. The transcript levels of more than 500 genes were more than 2.5-fold reduced in 35S:ARR1-SRDX transgenic seedlings, suggesting a broad function of B-type ARRs. By linking a SRDX-motif to the C-terminal of transcription activators, the chimeric gene has been changed into highly efficient negative regulons to repress the expression of target genes specifically and efficiently 31, 32.
Therefore, two CRISPR/dCas9-based transcriptional regulation vectors, CRISPR. The most pronounced phenotype was found in the arr1-3 arr10-5 arr12-1 triple loss-of-function mutant, which showed almost complete insensitivity to high levels of exogenously applied cytokinins. To identify novel transcription factors that function mainly in growth and development of Arabidopsis, we randomly chose and observed lines from an Arabidopsis CRES-T library, which is a pool of transgenic plants expressing SRDX-fused transcription factors driven by the 35S promoter. For TRD, three SRDX repression domains originated from the ERF transcription factor in tandem were selected duo to its effective repression of the target gene transcriptionally in plant (Heyl et al., 2008, Lowder et al., 2015, Piatek et al., 2015, Li et al., 2020). We employed a mutant-based approach to perform a detailed characterization of the roles of ARR1, ARR10, and ARR12 in plant growth and development. These results provide a new insight into the role of MsWRKY11 in regulating drought tolerance through lignin biosynthesis and stomatal formation in alfalfa.The type B Arabidopsis Response Regulators (ARRs) of Arabidopsis thaliana are transcription factors that act as positive regulators in the two-component cytokinin signaling pathway. By linking a SRDX-motif to the C-terminal of transcription activators, the chimeric gene has been changed into highly efficient negative regulons to repress the expression of target genes. MsWRKY22 was demonstrated to directly bind to W-box elements in MsWRKY11 promoter through in vitro and in vivo assays and positively activate MsWRKY11 expression. Lignin contents increased in the stem of MsWRKY11-OE plants under normal and drought conditions, but decreased in hairy roots of MsWRKY11-SRDX lines. Overview of attention for article published in Plant. The results showed that overexpression of MsWRKY11 in alfalfa decreased stomatal density in leaves and enhanced water use efficiency and drought tolerance. The transcript levels of more than 500 genes were more than 2.5-fold reduced in 35S: ARR1-SRDX transgenic seedlings, suggesting a broad function of B-type ARRs. The Transcriptional Repressor ARR1-SRDX Suppresses Pleiotropic Cytokinin Activities in Arabidopsis.
Its biological function and gene regulation mechanism were studied via overexpressed MsWRKY11 (MsWRKY11-OE) or dominantly repressed MsWRKY11 (MsWRKY11-SRDX) in alfalfa seedlings or hairy roots of alfalfa. In this study, an osmotic stress-responsive WRKY TF gene, named MsWRKY11, was cloned from alfalfa (Medicago sativa L.), which exhibits high expression level in stem and petiole. WRKY transcription factors are critical TFs involved in drought stress response. Drought stress limits the productivity of alfalfa which is one of the most important forage species around the world.
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