Comparison of mitochondrial gene expression and polysome loading in different tobacco tissues

Abstract

BackgroundTo investigate translational regulation of gene expression in plant mitochondria, a mitochondrial polysome isolation protocol was established for tobacco to investigate polysomal mRNA loading as a proxy for translational activity. Furthermore, we developed an oligonucleotide based microarray platform to determine the level of Nicotiana tabacum and Arabidopsis thaliana mitochondrial mRNA.ResultsMicroarray analysis of free and polysomal mRNAs was used to characterize differences in the levels of free transcripts and ribosome-bound mRNAs in various organs of tobacco plants. We have observed higher mitochondrial transcript levels in young leaves, flowers and floral buds as compared to fully expanded leaves and roots. A similar pattern of abundance was observed for ribosome-bound mitochondrial mRNAs in these tissues. However, the accumulation of the mitochondrial protein COX2 was found to be inversely related to that of its ribosome-bound mRNA.ConclusionsOur results indicate that the association of mitochondrial mRNAs to ribosomes is largely determined by the total transcript level of a gene. However, at least for Cox2, we demonstrated that the level of ribosome-bound mRNA is not reflected by the amount of COX2 protein.Electronic supplementary materialThe online version of this article (10.1186/s13007-017-0257-4) contains supplementary material, which is available to authorized users.

Particle bombardment-assisted peptide-mediated gene transfer for highly efficient transient assay

“…This plasmid contains the NLuc gene [12] driven by Prrn18. Prrn18 is the promoter region of the gene coding for mitochondrial 18S ribosomal RNA (rrn18) and probably activates strong transcription [13]. The DNA fragments containing Prrn18 was amplified using the primers, SacI-rrn18 F (5'-GCGAGCTCCGGAAGTAGCGCGA-CAAAGAG-3' , SacI site is underlined) and KpnI-rrn18 R (5'-GCGGTACCAACTCTTCTTTTGAGTATGAT-3' , KpnI site is underlined) from Nicotiana tabacum mitochondrial DNA as a template.…”

Effect of the overexpression of external alternative NADH dehydrogenase gene in Arabidopsis on the resistance of transformed tobacco plants to negative temperatures

“…Прямых Выделение митохондрий проводили по источнику [12] с модификациями. 80-100 г зеленых листьев табака растирали в охлажденной ступке в среде выделения (0,4 М сахароза, 50 мМ трис-HCl (рН 7,6), 1 мМ этиленгликольтетрауксусная кислота (ЭГТА), 10 мМ KH 2 PO 4 , 20 мМ аскорбат, 0,1%-й поливинилпирролидон, 0,5%-й бычий сывороточный альбумин (БСА), 0,5%-й цистеин).…”