scholarly journals Gene Inactivation by CRISPR-Cas9 in Nicotiana tabacum BY-2 Suspension Cells

2016 ◽  
Vol 7 ◽  
Author(s):  
Sébastien Mercx ◽  
Jérémie Tollet ◽  
Bertrand Magy ◽  
Catherine Navarre ◽  
Marc Boutry
Keyword(s):  
Nicotiana Tabacum ◽  
Gene Inactivation ◽  
Suspension Cells

Basket-shaped structures formed by F-actin in the nuclei of elongating cells of Nicotiana tabacum

1993 ◽  
Vol 71 (5) ◽  
pp. 725-731 ◽  
Author(s):  
Harry M. P. Kengen ◽  
Ton van Amstel ◽  
Bart Knuiman
Keyword(s):  
Nicotiana Tabacum ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Filamentous Actin ◽  
Suspension Cells ◽  
Confocal Immunofluorescence Microscopy ◽  
Cytoplasmic Actin ◽  
Extraction Buffer ◽  
Confocal Immunofluorescence

Phalloidin labelled with trimethyl rhodamine isothiocyanate in extraction buffer was used to stain actin in suspension cells of tobacco. Confocal immunofluorescence microscopy indicated the presence of filamentous actin containing structures in interphase nuclei of elongating cells of Nicotiana tabacum ‘Bright Yellow 2 Go’. The results were not affected by the omission of DMSO from the extraction medium. These structures, called actin baskets, were present in about 20% of the cells after induced elongation and varied in size, shape, and number per nucleus. The cytoplasmic actin array remained intact. It is proposed that the baskets have a transient character and are related to differentiation. Key words: confocal laser scanning microscopy, elongation, F-actin, nucleus, protoplasts, TRITC–phalloidin.

scholarly journals Enantioselective Introduction of Oxygen Functions into Alkenes. Oxidation of Limonene by the Cultured suspension Cells of Nicotiana tabacum.

1993 ◽  
Vol 10 (1) ◽  
pp. 89-91 ◽  
Author(s):  
Toshifumi HIRATA ◽  
Shunsuke IZUMI ◽  
Tomoko KIDO ◽  
Yoko SHIMOI ◽  
Yoshihiro IKEDA
Keyword(s):  
Nicotiana Tabacum ◽  
Suspension Cells

Stereoselectivity of the reduction of carvone and dihydrocarvone by suspension cells of Nicotiana tabacum

1982 ◽  
Vol 21 (9) ◽  
pp. 2209-2212 ◽  
Author(s):  
Toshifumi Hirata ◽  
Hiroki Hamada ◽  
Tadashi Aoki ◽  
Takayuki Suga
Keyword(s):  
Nicotiana Tabacum ◽  
Suspension Cells

Biochemical properties of the matrix metalloproteinase NtMMP1 from Nicotiana tabacum cv. BY-2 suspension cells

Planta ◽  
2010 ◽  
Vol 232 (4) ◽  
pp. 899-910 ◽  
Author(s):  
Manoj K. Mandal ◽  
Rainer Fischer ◽  
Stefan Schillberg ◽  
Andreas Schiermeyer
Keyword(s):  
Nicotiana Tabacum ◽  
Matrix Metalloproteinase ◽  
Biochemical Properties ◽  
Suspension Cells ◽  
The Matrix

Purification and biochemical characterization of NpABCG5/NpPDR5, a plant pleiotropic drug resistance transporter expressed in Nicotiana tabacum BY-2 suspension cells

2017 ◽  
Vol 474 (10) ◽  
pp. 1689-1703 ◽  
Author(s):  
Frédéric Toussaint ◽  
Baptiste Pierman ◽  
Aurélie Bertin ◽  
Daniel Lévy ◽  
Marc Boutry
Keyword(s):  
Electron Microscopy ◽  
Drug Resistance ◽  
Nicotiana Tabacum ◽  
Abc Transporters ◽  
Plasma Membranes ◽  
Biochemical Characterization ◽  
Specific Activity ◽  
Pleiotropic Drug Resistance ◽  
Suspension Cells

Pleiotropic drug resistance (PDR) transporters belong to the ABCG subfamily of ATP-binding cassette (ABC) transporters and are involved in the transport of various molecules across plasma membranes. During evolution, PDR genes appeared independently in fungi and in plants from a duplication of a half-size ABC gene. The enzymatic properties of purified PDR transporters from yeast have been characterized. This is not the case for any plant PDR transporter, or, incidentally, for any purified plant ABC transporter. Yet, plant PDR transporters play important roles in plant physiology such as hormone signaling or resistance to pathogens or herbivores. Here, we describe the expression, purification, enzymatic characterization and 2D analysis by electron microscopy of NpABCG5/NpPDR5 from Nicotiana plumbaginifolia, which has been shown to be involved in the plant defense against herbivores. We constitutively expressed NpABCG5/NpPDR5, provided with a His-tag in a homologous system: suspension cells from Nicotiana tabacum (Bright Yellow 2 line). NpABCG5/NpPDR5 was targeted to the plasma membrane and was solubilized by dodecyl maltoside and purified by Ni-affinity chromatography. The ATP-hydrolyzing specific activity (27 nmol min−1 mg−1) was stimulated seven-fold in the presence of 0.1% asolectin. Electron microscopy analysis indicated that NpABCG5/NpPDR5 is monomeric and with dimensions shorter than those of known ABC transporters. Enzymatic data (optimal pH and sensitivity to inhibitors) confirmed that plant and fungal PDR transporters have different properties. These data also show that N. tabacum suspension cells are a convenient host for the purification and biochemical characterization of ABC transporters.

scholarly journals Evolutionary and Predictive Functional Insights into the Aquaporin Gene Family in the Allotetraploid Plant Nicotiana tabacum

2020 ◽  
Vol 21 (13) ◽  
pp. 4743 ◽  
Author(s):  
Jahed Ahmed ◽  
Sébastien Mercx ◽  
Marc Boutry ◽  
François Chaumont
Keyword(s):  
Nicotiana Tabacum ◽  
Substrate Specificity ◽  
Gene Family ◽  
Splice Variants ◽  
Nicotiana Sylvestris ◽  
Membrane Diffusion ◽  
Suspension Cells ◽  
Aquaporin Gene ◽  
Organ Specific ◽  
Tabacum Plant

Aquaporins (AQPs) are a class of integral membrane proteins that facilitate the membrane diffusion of water and other small solutes. Nicotiana tabacum is an important model plant, and its allotetraploid genome has recently been released, providing us with the opportunity to analyze the AQP gene family and its evolution. A total of 88 full-length AQP genes were identified in the N. tabacum genome, and the encoding proteins were assigned into five subfamilies: 34 plasma membrane intrinsic proteins (PIPs); 27 tonoplast intrinsic proteins (TIPs); 20 nodulin26-like intrinsic proteins (NIPs); 3 small basic intrinsic proteins (SIPs); 4 uncharacterized X intrinsic proteins (XIPs), including two splice variants. We also analyzed the genomes of two N. tabacum ancestors, Nicotiana tomentosiformis and Nicotiana sylvestris, and identified 49 AQP genes in each species. Functional prediction, based on the substrate specificity-determining positions (SDPs), revealed significant differences in substrate specificity among the AQP subfamilies. Analysis of the organ-specific AQP expression levels in the N. tabacum plant and RNA-seq data of N. tabacum bright yellow-2 suspension cells indicated that many AQPs are simultaneously expressed, but differentially, according to the organs or the cells. Altogether, these data constitute an important resource for future investigations of the molecular, evolutionary, and physiological functions of AQPs in N. tabacum.

Different subcellular localization and glycosylation for a functional antibody expressed in Nicotiana tabacum plants and suspension cells

2009 ◽  
Vol 18 (3) ◽  
pp. 467-482 ◽  
Author(s):  
Benoit De Muynck ◽  
Catherine Navarre ◽  
Yannick Nizet ◽  
Johannes Stadlmann ◽  
Marc Boutry
Keyword(s):  
Nicotiana Tabacum ◽  
Subcellular Localization ◽  
Suspension Cells

scholarly journals Alamethicin permeabilizes the plasma membrane and mitochondria but not the tonoplast in tobacco (Nicotiana tabacum L. cv Bright Yellow) suspension cells

2005 ◽  
Vol 389 (3) ◽  
pp. 695-704 ◽  
Author(s):  
Sandra Matic ◽  
Daniela A. Geisler ◽  
Ian M. Møller ◽  
Susanne Widell ◽  
Allan G. Rasmusson
Keyword(s):  
Nicotiana Tabacum ◽  
Plasma Membranes ◽  
Animal Cells ◽  
Suspension Cells ◽  
Respiratory Enzymes ◽  
Mitochondrial Oxygen Consumption ◽  
Permeabilized Cells ◽  
Bright Yellow ◽  
Transport Chain ◽  
Nicotiana Tabacum L

The ion channel-forming peptide AlaM (alamethicin) is known to permeabilize isolated mitochondria as well as animal cells. When intact tobacco (Nicotiana tabacum L.) Bright Yellow-2 cells were treated with AlaM, the cells became permeable for low-molecular-mass molecules as shown by induced leakage of NAD(P)+. After the addition of cofactors and substrates, activities of cytosolic as well as mitochondrial respiratory enzymes could be directly determined inside the permeabilized cells. However, at an AlaM concentration at which the cytoplasmic enzymes were maximally accessible, the vacuole remained intact, as indicated by an unaffected tonoplast proton gradient. Low-flux permeabilization of plasma membranes and mitochondria at moderate AlaM concentrations was reversible and did not affect cell vigour. Higher AlaM concentrations induced cell death. After the addition of catalase that removes the H2O2 necessary for NADH oxidation by apoplastic peroxidases, mitochondrial oxygen consumption could be measured in permeabilized cells. Inhibitor-sensitive oxidation of the respiratory substrates succinate, malate and NADH was observed after the addition of the appropriate coenzymes (ATP, NAD+). The capacities of different pathways in the respiratory electron-transport chain could thus be determined directly. We conclude that AlaM permeabilization provides a very useful tool for monitoring metabolic pathways or individual enzymes in their native proteinaceous environment with controlled cofactor concentrations. Possible uses and limitations of this method for plant cell research are discussed.

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