BY-2 tobacco

BY-2 tobacco culture was propagated as previously described. Cells were harvested 1-12 d after sub-culturing, and used for experiments after addition of 10 /tM CLA. LaCla solution (various concentration) was added to cell suspension in glass tubes placed in a CHEM-GLOW Photometer (AMINCO., Silver Spring, MD, USA), and OXB was monitored by CLA-CL, and expressed as relative CL units (rcu) as previously described. Cells were treated with ZnS04 (0.3, 1, 3 mM), 2 min... 

Kawano T, Kawano N, Muto S, Lapeyrie F. Retardation and inhibition of the cation-induced superoxide generation in BY-2 tobacco cell suspension culture by Zn and Mn Physiol Plantar, 2002 114 395-404. 

R.P. Sabba, N.A. Durso, K.C. Vaughn. Structural and immunocytochemical characterization of the walls of dichlorobenil-adapted BY-2 tobacco cells. Int J Plant Sci, 1999, 160, 275-290... 

Completely different mechanisms are involved in the self-assembly of the tobacco mosaic virus (TMV). This virus consists of single-strand RNA, which is surrounded by 2,130 identical protein units, each of which consists of 158 amino acid residues. A virus particle, which requires the tobacco plant as a host, has a rodlike structure with helical symmetry ( Stanley needles ). It is 300 nm long, with a diameter of 18nm. The protein and RNA fractions can be separated, and the viral... 

Gerber E, Hemmerlin A, Hartmann M, Heintz D, Hartmann MA, Mutterer J, Rodriguez-Concepcion M, Boronat A, Van Dorsselaer A, Rohmer M, CroweU DN, Bach TJ (2009) The plastidial 2-C-methyl-D-erythritol 4-phosphate pathway provides the isoprenyl moiety for protein geranylgeranylation in tobacco BY-2 ceUs. Plant Cell 21 285-300... 

Weight loss. Smoke of the leaf, administered by inhalation to mice at variable doses twice daily for 5 days followed by 2 nontreatment days, dosed for two to six cycles, was active b Tobacco smoke, administered to 3-week-old Wistar rats on a vitamin E-depleted or normal diet, for 4 weeks, significantly suppressed body weight increases, particularly in the vitamin E-depleted group . 

Molina, A., Veramendi, J., and Hervas-Stnbbs, S. (2005). Indnction of nentral-izing antibodies by a tobacco chloroplast-derived vaccine based on a B cell epitope from canine parvovirns. Virology 342(2) 266-275. 

D also Interacts with lAA to reverse glyphosate growth Inhibition In tissue cultures (l i, 1701. but not In Intact plants (121) Extractable PAL activity In soybean seedlings was Inhibited by 2,4-D (1711 and 2,4-D lowered PAL activity Increases caused by glyphosate (1711. The herbicide has also been examined for Interactions of Induction and secretion of pathogenesis-related proteins In tobacco suspension cultures (1721. 

Hemmerlin, A., Gerber, E., Feldtrauer, J.R, Wentzinger, L., Hartmann, M.A., Tritsch, D., Hoeffler, J.F., Rohmer, M. and Bach, T.J. (2004) A review of tobacco BY-2 cells as an excellent system to study the synthesis and function of sterols and other isoprenoids. Lipids, 39, 723-35. 

Recently, a photoactivatable variant from Aequoria victoria green fluorescent protein (pa-GFP) was reported (Patterson and Lippincott-Schwartz 2002), yielding an increase in fluorescence emission intensity (at k 520 nm) by a factor of 100 when excited at k 488 nm after spectral activation at A. 408 nm. This phenomenon is due to an internal photoconversion process in the protein and allows spectral photoactivation of this protein in a very local way such as in the nucleus of a living cell (Post et al. 2005). In tobacco BY-2 protoplasts, we transiently co-expressed pa-GFP or pa-GFP fusion proteins and red-fluorescent protein (DsRed)-tagged prenylated Rab acceptor 1 (Pral At2g38360), a membrane protein that localizes in speckles around the nuclear envelope. The DsRed transfection allows proper cell identification and visualization before activation (via Pral -DsRed fluorescence). After pa-GFP... 

Figure 6. Dynamics of free diffusion of pa-GFP in a live protoplast. Five selected IP-transmission fluorescence images of a tobacco BY-2 protoplast expressing pa-GFP (A) at the onset of the experiment before activation, (B) during 2P-activation of pa-GFP, and (C-E) after 2P-activation were taken at the time points indicated. (A) Before 2P-activation of pa-GFP in the nucleus (dotted line) the average fluorescence intensity is barely detectable. 2P-activation of pa-GFP was initiated with a fs-laser burst of 3 s covering an area of 7 x 8 p,m with 4 parallel laser foci (10 mW at 800 nm per focus). (B) Shortly after photo-activation a strong fluorescence signal was detected and (C-E) the diffusion of photo-activated pa-GFP from the nucleus into the cytoplasm was monitored until equilibrium of partitioning between the two cellular compartments was reached. Fluorescence intensity scales are shown in each panel to the left.

Figure 7. pa-GFP dynamics by parallel 2P-epifluorescence microscopy (64 foci, 920 nm, 240 mW) in a tobacco BY-2 protoplast. Quantitative analysis of the decrease of nuclear pa-GFP 2P-epifluorescence, giving a diffusion time constant of 123 s. 

Hollweg et al.26 determined simultaneously nicotine and water in tobacco smoke condensates. For the nicotine determination a packed column, 1.8 m long by 2 mm I.D. with 10 % Carbowax 20 M on Chromosorb WHP 60-80 mesh was used. Since the column was prepared without addition of... 

NONCOMPETITIVE INHIBITION OF LANTHANIDE-INDUCED OXIDATIVE BURST BY ZINC IN TOBACCO BY-2 CELLS ... 

Kawano T, Kadono T, Furuichi T, Muto S, Lapeyrie F. Aluminum-induced distortion in calcium signaling involving oxidative bursts and channel regulations in tobacco BY-2 cells. Biochem Biophys Res Commun 2003 308 35-42. 

Suspension cell cultures have been derived from a number of different plant species, including the widely-used laboratory model Arabidopsis thaliana [15], plants such as Catharanthus roseus and Taxus cus-pidata which are used to produce valuable secondary metaboHtes [16, 17], and important domestic crops such as tobacco, rice, alfalfa, tomato, and soybean [18-22]. Because cell lines from domestic crop species are well-characterized, they have been the most frequently used for recombinant protein production. The most popular cell hnes include those derived from the tobacco cultivars Bright Yellow 2 (BY-2) (Fig. 9.1) and Nicotiana tabacum 1 (NT-1) [2]. 

Fig. 9.1 Tobacco BY-2 suspension cells (A original magnification x400) are cultivated in shake flasks (B) and bioreactors (C)...

Tobacco cv BY-2 BiscFv antibody Enhanced Cytosolic (at detection 40... 

Tobacco cv BY-2 suspension cells MAb against HBsAg CaMV 35 S Secreted, ca. 50/50 between supernatant and cells, total max ca. 15 mg 49... 

Tobacco cv BY-2 suspension cells Desmodus rotundus Salivary plasminogen activator al (DSPAal) Enhanced CaMV35S Intracellular, up to 1.5 pg g FW and degraded when secreted to the supernatant (3 different signal peptides were used) 50... 

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