The brassinosteroid (BR) phytohormone is an important regulator of plant growth.
#Srdx domain Activator
Based on the findings of this study, we were thus able to establish the mechanisms whereby the transcriptional activator CmBBX22 negatively regulates drought tolerance in chrysanthemum via the regulation of ABA response, stomatal conductance and antioxidant responses. Transcriptome analysis and physiological measurements indicated the potential involvement of CmBBX22-mediated ABA response, stomatal conductance and antioxidant responses in the negative regulation of drought tolerance in chrysanthemum. Overexpression of CmBBX22 in chrysanthemum was found to reduce plant drought tolerance, whereas expression of the chimeric repressor CmBBX22-SRDX was found to promote a higher drought tolerance than that shown by wild-type plants, indicating that CmBBX22 negatively regulates drought tolerance in chrysanthemum. Subcellular localization and transactivation assay analyses revealed that CmBBX22 was localized in nucleus and possessed transactivation activity. Here, we cloned CmBBX22 and further determined the function of CmBBX22 in response to drought treatment. Although BBX proteins have been studied in great detail in the model plant Arabidopsis, their roles in crop plants such as chrysanthemum are still largely uninvestigated. 35S, cauliflower mosaic virus 35S promoter W, the translational enhancer sequence from tobacco mosaic virus TNOS, nopaline synthase terminator THSP, Arabidopsis HSP18.2 terminator HPT, hygromycin B phosphotransferase RB, right border LB, left border CmR, chloramphenicol-resistant marker ccd B, negative selection marker.īBX transcription factors play vital roles in plant growth, development, and stress responses. The promoter and TF are inserted into R4pGWB5_SRDX by an LR reaction. The entry clone of the promoter is constructed by a BP reaction between pDONRG-P4P1R and a PCR-amplified promoter with att B4 and att B1r sequences. (C) Multisite Gateway vector system (R4pGWB- SRDX series). The TF entry clone is constructed by a BP reaction between pDONR-207 and a PCR-amplified coding sequence of a TF without a stop codon with att B1 and att B2 sequences, and inserted into pDEST_35S_SRDX_BCKH by an LR reaction. (B) 35S-driven Gateway vector system (pDEST_35S_SRDX_BCKH series). Step2: The transgene cassette on the entry clone is transferred into pBCKH by an LR reaction. Step1: The coding sequence of a PCR- amplified TF without a stop codon is cloned via blunt-end ligation into the SmaI site of p35SSRDXG to fuse with SRDX. Overview of CRES-T vector systems and their derivatives. We, therefore, propose that the appropriate CRES-T vector should be chosen depending on situations and purposes. We found that the HSP terminator increased transcription efficiency or transcript stability in contrast, these factors were negatively affected by the Gateway linker sequence in our vector system. However, the HSP terminator compensated for the negative effect of the Gateway sequence and improved the efficiency of CRES-T in all cases tested and resulted in the highest efficiency achieved to date. Our test experiments revealed that the CRES-T vector containing the Gateway linker sequence within the transcribed region showed reduced efficiency of CRES-T when compared with the traditional CRES-T vector. In this study, we developed new CRES-T vectors that are efficient and convenient to use by employing the Gateway system, a new vector backbone and a terminator derived from the heat shock protein 18.2 (HSP) gene. However, the traditional CRES-T vectors are inconvenient for gene cloning and promoter exchange. For CRES-T, a transcription factor is converted to a strong repressor by fusion with an SRDX repression domain, which is then expressed in plants to induce a loss-of-function phenotype. Chimeric repressor gene-silencing technology (CRES-T) is a powerful tool that has recently been developed for the functional analysis of plant transcription factors and for the genetic manipulation of plant traits.
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