Project description:Nicotiana benthamiana is a widely used organism for expression of foreign proteins. To increase protein abundance, overexpression promoters have been developed. However, several proteins cause negative effects on plant growth and development when they are overexpressed. Therefore, overexpression promoters cannot be widely used to generate transgenic plants that produce foreign proteins. In this study, we tried to find stress-inducible promoters which specifically express downstream genes when plants were exposed to stress conditions. By RNA-sequencing analysis of heat- and wound-treated plants, we found that NbHSP20 promoters can be used as a heat-inducible promoters to express foreign proteins in N. benthamiana.
Project description:We assessed the off-target effects of M2 expression on the leaf proteome of Nicotiana benthamiana infiltrated with the bacterial gene vector Agrobacterium tumefaciens. A iTRAQ quantitative proteomics procedure was followed to compare the leaf proteomes of plants agroinfiltrated with either an ‘empty’ vector or an M2-encoding vector. Leaves infiltrated with the empty vector had a low soluble protein content compared to non-infiltrated control leaves, associated with increased levels of stress-related proteins but decreased levels of photosynthesis-associated proteins. M2 expression partly compromised these effects of agroinfiltration to restore soluble protein content in leaf tissue, associated with restored levels of photosynthesis-associated proteins and reduced levels of stress-related proteins in the apoplast. These data illustrate the cell-wide influence of Golgi lumen pH homeostasis on the leaf proteome of N. benthamiana responding to microbial challenge.
Project description:Nonsense mediated decay (NMD) is a translation termination coupled quality control system. NMD identifies and degrades aberrant mRNAs containing premature termination codons, thereby preventing the accumulation of detrimental truncated proteins. NMD also controls the expression of several normal mRNAs and non-coding transcripts. RNA-seq assays were conducted to identify the NMD regulated genes in Arabidopsis. However, NMD targets have not been studied in other plants. To identify the conserved NMD targets, we wanted to identify the NMD regulated genes in Nicotiana benthamiana model species. Virus induced gene silencing was used to inactivate UPF1 NMD key factor in N. benthamiana, and then comparative RNA-seq experiment was carried out using UPF1-silenced and control–silenced plants. We found that in N. benthamiana significantly more genes are controlled by NMD than in Arabidopsis and that, surprisingly few conserved NMD targets can be recognized. These results suggest that while the mechanism of NMD is highly conserved, the set of NMD controlled genes can be quickly changed in plants.
Project description:Gene expression changes between wildtype (WT) Nicotiana benthamiana plants with and without infection with Potato Spindle Tuber Viroid (PSTVd)