Project description:WIPK and SIPK, two tobacco MAPKs, are rapidly activated by wounding. Goal is to identify the genes regulated by WIPK and SIPK. For this purpose, we produced transgenic tobacco plants in which expression of WIPK and SIPK (WIPK/SIPK-suppressed plants) is suppressed by RNAi.
Project description:WIPK and SIPK, two tobacco MAPKs, are rapidly activated by wounding. Goal is to identify the genes regulated by WIPK and SIPK. For this purpose, we produced transgenic tobacco plants in which expression of WIPK and SIPK is suppressed by RNAi (WIPK/SIPK-suppressed plants).
Project description:Agilent 4x44k tobacco micro array of wild type tobacco (WT) and whole tobacco mosaic virus (TMV) containing transgenic tobacco plants. The transgenic plants before resistance break (BRB-6 weeks), after resistance break (ARB-8 weeks) and wild type tobacco plants infected with TMV (TMVi-9weeks) leaves were analyzed. Three biological replicates were performed for each sample.
Project description:Histone acetylation is important for the transcriptional regulation in the ethylene response and the nuclear metabolic production of acetyl coenzyme A (acetyl-CoA) plays key roles in histone acetylation. But how the nucleus produces acetyl-CoA for histone acetylation and transcription regulation in the ethylene response is completely unknown. Here we show that functional pyruvate dehydrogenase complex (PDC) translocates from the mitochondria to the nucleus in response to ethylene, generating nuclear acetyl CoA from pyruvate to regulate EIN2-directed histone acetylation and transcription in the target genes. It has been reported that the dephosphorylated E1 subunit of the PDC complex is required for the PDC activity, we examined the phosphorylation status change of E1 in the nucleus in response to ethylene. The MS data shows quantitative phosphorylation differences between cytosolic and nuclear fraction supporting the dephosphorylation of E1 upon ethylene treatment.
Project description:Transgenic tobacco plants expressing begomoviral AC2 RNA silencing suppressor were used to compare transcriptional changes in the transcriptome between transgenic and wild type tobacco plants. Transcriptional analysis using Agilent 4x44k tobacco array was performed of six week-old leaves and 3-5 month-old flowers taken from the same plants as leaf samples.
Project description:Purpose: Trichomes, developing from the epidermis of nearly all terrestrial plants, provide good structural resistance against insect herbivores and an excellent model for studying the molecular mechanisms underlying cell fate determination. Regulation of trichomes in Rosids has been well characterized. However, little is known about the cell proliferation molecular processes during multicellular trichome formation in Asterids. Methods: The transcriptomes of between Wov transgenic and wild-type tobacco by RNA-seq analysis were evaluated using the Illumina HiSeq™ 2000 sequencing platform. Raw sequences were filtered and the resulting sets of clean reads were used for the following analysis by Tophat and DEGseq software. qRT–PCR validation was performed using SYBR Green assays. Results: In this study, we identified two point mutations in a novel allele (Wov) at Wo locus. Ectopic expression of Wov in tobacco and potato induces much more trichome formation than wild type. To gain new insights into the underlying mechanisms during the processes of these trichomes formation, we compared the gene expression profiles between Wov transgenic and wild-type tobacco by RNA-seq analysis. A total of 544 co-DEGs were detected between transgenic and wild-type tobacco. Functional assignments of the co-DEGs indicated that 33 reliable pathways are altered in transgenic tobacco plants. The most noticeable pathways are fatty acid metabolism, amino acid biosynthesis and metabolism, and plant hormone signal transduction. Results suggest that these enhanced processes are critical for the cell proliferation during multicellular trichome formation in transgenic plants. In addition, the transcriptional levels of homologues of trichome regulators in Rosids were not significantly changed, whereas homologues of genes (Wo and SlCycB2) in Asterids were significantly upregulated in Wov transgenic tobacco plants. Conclusions: This study presents a global picture of the gene expression changes induced by Wov- gene in tobacco. And the results provided us new insight into the molecular processes controlling multicellular formation in tobacco. Furthermore, we inferred that trichomes in solanaceous species might share a common network.
Project description:Agilent 4x44k tobacco micro array of wild type tobacco, empty vector control, and HC-Pro transgenic tobacco plants. Both 1-month old leaves and flowers were analyzed. Three biological replicates were performed of each sample.
Project description:Purpose: Trichomes, developing from the epidermis of nearly all terrestrial plants, provide good structural resistance against insect herbivores and an excellent model for studying the molecular mechanisms underlying cell fate determination. Regulation of trichomes in Rosids has been well characterized. However, little is known about the cell proliferation molecular processes during multicellular trichome formation in Asterids. Methods: The transcriptomes of between Wov transgenic and wild-type tobacco by RNA-seq analysis were evaluated using the Illumina HiSeq™ 2000 sequencing platform. Raw sequences were filtered and the resulting sets of clean reads were used for the following analysis by Tophat and DEGseq software. qRT–PCR validation was performed using SYBR Green assays. Results: In this study, we identified two point mutations in a novel allele (Wov) at Wo locus. Ectopic expression of Wov in tobacco and potato induces much more trichome formation than wild type. To gain new insights into the underlying mechanisms during the processes of these trichomes formation, we compared the gene expression profiles between Wov transgenic and wild-type tobacco by RNA-seq analysis. A total of 544 co-DEGs were detected between transgenic and wild-type tobacco. Functional assignments of the co-DEGs indicated that 33 reliable pathways are altered in transgenic tobacco plants. The most noticeable pathways are fatty acid metabolism, amino acid biosynthesis and metabolism, and plant hormone signal transduction. Results suggest that these enhanced processes are critical for the cell proliferation during multicellular trichome formation in transgenic plants. In addition, the transcriptional levels of homologues of trichome regulators in Rosids were not significantly changed, whereas homologues of genes (Wo and SlCycB2) in Asterids were significantly upregulated in Wov transgenic tobacco plants. Conclusions: This study presents a global picture of the gene expression changes induced by Wov- gene in tobacco. And the results provided us new insight into the molecular processes controlling multicellular formation in tobacco. Furthermore, we inferred that trichomes in solanaceous species might share a common network. The transcriptomes of between Wov transgenic and wild-type tobacco by RNA-seq analysis were evaluated, in duplicate, using the Illumina sequencing platform.