Project description:Primary cell wall is an essential cell structure for plant playing major roles in plant growth, differentiation, and stress responses. Here we demonstrate that a group of AP2-ERF transcription factor regulates primary cell wall formation and can induce massive accumulation of it in empty fiber cell of the nst1-1 nst3-1 mutant lacking secondary cell wall in Arabidopsis. The transgenic plants expressing one of the AP2-ERF transcription factors fused with VP16 transcriptional activation domain under the control of NST3 promoter in the nst1-1 nst3-1 mutant showed similar level of cell wall contents with wild type by the massive accumulation of cell wall which lacks lignin and xylan. The transgenic plants showed 70% higher saccharification efficiency than wild type. Gene expression analysis using microarray revealed that genes related to primary cell wall were highly upregulated in the transgenic plant. Moreover, chimeric-activator of the AP2-ERF transcription factor accelerated cell wall regeneration of mesophyll protoplast of Arabidopsis while the chimeric-repressor retarded it. These data suggest that the group of AP2-ERF transcription factor is key regulator of the primary cell wall formation in plant and could be employed to produce massive cell wall with readily extractable feature.
Project description:Wood is one of the most important and enormous biomass that is widely used in our life. It is formed by successive addition of secondary xylem that develops continuously from cambium. The transcriptome of nst1 nst3 âdouble-knockoutâ lines was examined to know the effect of mutations on wood formation. Experiment Overall Design: Total RNA was extracted from the base 4 cm part of inflorescence stems whose heights were between 12 and 17 cm of three independent nst1-1 nst3-1 double T-DNA tagged lines and the whole transcriptome was compared with that of wild-type plant.
Project description:The Arabidopsis thaliana NAC domain transcription factor, VASCULAR-RELATED NAC-DOMAIN7 (VND7), acts as a key regulator of xylem vessel differentiation. In order to identify direct target genes of VND7, we performed global transcriptome analysis using Arabidopsis transgenic lines in which VND7 activity could be induced post-translationally. This analysis identified 63 putative direct target genes of VND7, which encode a broad range of proteins, such as transcription factors, IRREGULAR XYLEM proteins and proteolytic enzymes, known to be closely associated with xylem vessel formation. Recombinant VND7 protein binds to several promoter sequences present in candidate direct target genes: specifically, in the promoter of XYLEM CYSTEINE PEPTIDASE1, two distinct regions were demonstrated to be responsible for VND7 binding. We also found that expression of VND7 restores secondary cell wall formation in the fiber cells of inflorescence stems of nst1 nst3 double mutants, as well as expression of NAC SECONDARY WALL THICKENING PROMOTING FACTOR3 (NST3, however, the vessel-type secondary wall deposition was observed only as a result of VND7 expression. These findings indicated that VND7 upregulates, directly and/or indirectly, many genes involved in a wide range of processes in xylem vessel differentiation, and that its target genes are partially different from those of NSTs.
Project description:Wood is one of the most important and enormous biomass that is widely used in our life. It is formed by successive addition of secondary xylem that develops continuously from cambium. The transcriptome of nst1 nst3 “double-knockout” lines was examined to know the effect of mutations on wood formation. Keywords: mutant vs wt comparison
Project description:In plants, secondary wall thickenings play important roles in various biological processes, although the factors regulating these processes remain to be characterized. We show that expression of chimeric repressors derived from NAC SECONDARY WALL THICKENINGS PROMOTING FACTOR1 (NST1) and NST2 in Arabidopsis resulted in an anther dehiscence defect due to loss of secondary wall thickening in anther endothecium. Plants with double, but not single, T-DNA-tagged lines for NST1 and NST2, had the same anther-indehiscent phenotype as transgenic plants that expressed the individual chimeric repressors, indicating that NST1 and NST2 are redundant in regulating secondary wall thickening in anther walls. The activity of the NST2 promoter was particularly strong in anther tissue, while that of the NST1 promoter was detected in various tissues in which lignified secondary walls develop. Ectopic expression of NST1 or NST2 induced ectopic thickening of secondary walls in various above-ground tissues. Epidermal cells with ectopic thickening of secondary walls had structural features similar to those of tracheary elements. However, among genes involved in the differentiation of tracheary elements, only those related to secondary wall synthesis were clearly upregulated. None of the genes involved in programmed cell death was similarly affected. Our results suggest NAC transcription factors as possible regulators of secondary wall thickening in various tissues. Keywords: transgenic vs wt comparison
Project description:In plants, secondary wall thickenings play important roles in various biological processes, although the factors regulating these processes remain to be characterized. We show that expression of chimeric repressors derived from NAC SECONDARY WALL THICKENINGS PROMOTING FACTOR1 (NST1) and NST2 in Arabidopsis resulted in an anther dehiscence defect due to loss of secondary wall thickening in anther endothecium. Plants with double, but not single, T-DNA-tagged lines for NST1 and NST2, had the same anther-indehiscent phenotype as transgenic plants that expressed the individual chimeric repressors, indicating that NST1 and NST2 are redundant in regulating secondary wall thickening in anther walls. The activity of the NST2 promoter was particularly strong in anther tissue, while that of the NST1 promoter was detected in various tissues in which lignified secondary walls develop. Ectopic expression of NST1 or NST2 induced ectopic thickening of secondary walls in various above-ground tissues. Epidermal cells with ectopic thickening of secondary walls had structural features similar to those of tracheary elements. However, among genes involved in the differentiation of tracheary elements, only those related to secondary wall synthesis were clearly upregulated. None of the genes involved in programmed cell death was similarly affected. Our results suggest NAC transcription factors as possible regulators of secondary wall thickening in various tissues. Experiment Overall Design: Total RNA was extracted from rosette leaves of two independent 2-week-old T1 plants over-expressing NST1 driven by 35S promoter and the whole transcriptome was compared with that of wild-type plant.
Project description:In this study, the genes that encode AP2/ERF transcription factors, namely OpERF1 to OpERF5, were isolated from HR of O. pumila. Phylogenetic analysis of AP2/ERF protein sequences suggested the close evolutionary relationship of OpERF1 with stress-responsive ERF factors in Arabidopsis and of OpERF2 with ERF factors reported to regulate alkaloid production, such as ORCA3 in Catharanthus roseus, NIC2-locus ERFs in tobacco, and JRE4 in tomato. We generated the HR lines of O. pumila, ERF1i and ERF2i, in which the expression of OpERF1 and OpERF2, respectively, was suppressed using RNA interference technique. The transcriptome and metabolome of these suppressed HR were analyzed for functional characterization of OpERF1 and OpERF2.