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

Hameed, Muhammad Waqar; Juszczak, Ilona; Bock, Ralph; Dongen, Joost T. van

doi:10.1186/s13007-017-0257-4

Cited by 4 publications

(2 citation statements)

References 89 publications

(98 reference statements)

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“…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.…”

Section: Plasmid Constructionmentioning

confidence: 99%

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

et al. 2023

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Objective A centrifugation-assisted peptide-mediated gene transfer (CAPT) method was recently developed as an efficient system for gene delivery into plant cells. However, the gene transfer efficiency of CAPT into plant cells was not entirely satisfactory for detecting transient expression of a transgene driven into mitochondria. Here, we report a new gene delivery system using a method called particle bombardment-assisted peptide-mediated gene transfer (PBPT). Results We investigated various parameters of the PBPT method to increase transient gene expression efficiency in Brassica campestris. The optimal conditions for PBPT were a single bombardment with gold particles coated with a DNA‒peptide complex (6 µg of DNA and 2 µg of peptide) at an acceleration pressure of 5 kg/cm2 and a target distance of 12.5 cm. Moreover, bombardment under the optimal conditions successfully transferred the transgene into the cells of other plant species, namely B. juncea and tomato. Thus, we developed a PBPT method for highly efficient delivery of a DNA‒peptide complex into plant mitochondria.

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Section: Plasmid Constructionmentioning

confidence: 99%

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

et al. 2023

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“…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).…”

Section: Introductionunclassified

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

Borovskii,

Gorbyleva,

Katyshev

et al. 2023

Izvestiâ vuzov. Prikladnaâ himiâ i biotehnologiâ

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The study aims to confirm the functional activity and localization of the At_NDB2 transgenic protein of Arabidopsis in tobacco cells and to evaluate the effect produced by the permanent expression of external alternative NADH dehydrogenase on the resistance of a heat-loving plant to low temperatures. Proteins and mitochondria were isolated from the leaves of tobacco plants grown at 25 °С (day/night) at the 7-leaf stage. At_NDB2 protein localization in mitochondria was determined via electrophoresis and immunoblotting. The functional activity of At_NDB2 was confirmed through the polarography of isolated mitochondria and the specific mitochondrial complex I inhibitor (rotenone). It was also found that the cyanide-resistant respiration rate and the activity of an alternative oxidase enzyme were significantly higher in transgenic plants than in wild-type plants. In order to determine the resistance to low temperatures, the parent and transgenic tobacco plants were grown in soil until the 2–3 and 6–7 leaf stages, after which they were kept at 3 to -3 °С for a day in the dark and left to regrow at 25 °С for seven days. Although it was previously shown that oxidative stress is reduced in transgenic plants at low temperatures as compared to wild-type plants, it was established that the tolerance of transgenic and nontransformed plants does not differ. Thus, alternative NADH dehydrogenase activity was found to reduce oxidative stress and increase alternative oxidase activity, without enhancing the resistance of Nicotiana tabacum to negative temperatures.

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Experimental approaches to studying translation in plant semi-autonomous organelles

Kwasniak-Owczarek,

Janska

2024

Journal of Experimental Botany

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Plant mitochondria and chloroplasts are semi-autonomous organelles originated from free-living bacteria and retaining respective reduced genomes during evolution. As a consequence, relatively few of the mitochondrial and chloroplast proteins are encoded in the organellar genomes and synthesized by the organellar ribosomes. Since the both organellar genomes encode mainly components of the energy transduction systems, oxidative phosphorylation in mitochondria and photosynthetic apparatus in chloroplasts, understanding the organellar translation is critical to a thorough comprehension of the key aspects of mitochondrial and chloroplast activity affecting plant growth and development. Recent studies have clearly shown that translation is a key regulatory node in the expression of plant organellar genes, underscoring the need for an adequate methodology to study this unique stage of gene expression. The organellar translatome can be analysed by studying newly synthesized proteins or the mRNA pool recruited to the organellar ribosomes. In this review, we present in some detail the experimental approaches used to date for studying translation in the plant bioenergetic organelles. Their benefits and limitations, as well as the critical steps are discussed. Additionally, we briefly mention several recently developed strategies to study organellar translation that have not yet been applied to plants.

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

Cited by 4 publications

References 89 publications

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

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

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

Experimental approaches to studying translation in plant semi-autonomous organelles

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