Tobacco protoplasts

Nature of the ZIO Reaction Effect of pH in the ZIO Mixture Effect of Temperature of the ZIO Mixture Summary of the ZIO Technique in Visualizing the Golgi Apparatus Tobacco Protoplast-ZIO Staining References... 

The ER, Golgi bodies, and nuclear envelopes in cotyledon cells and cells of Chara corallina showed a marked increase in density by ZIO staining (38,39). We investigated the continuity of the Golgi apparatus into the ER of cultured tobacco protoplasts by ZIO staining (40). 

Kaneko TS, Sato M, Watanabe R, Konomi M, Osumi M, Takatsuki A. Observation of the continuity of Golgi apparatus with endoplasmic reticulum induced by bre-feldin A studied by zinc iodide-osmium staining in tobacco protoplasts. J Electron Microsc 1998 47 93-97. 

NT474 Zelcer, A. and E. Galun. Culture of NT483 newly isolated tobacco protoplasts precursor incorporation into protein,... 

Frigerio, L., Vitale, A., Lord, J.M., Ceriotti, A. and Roberts, L.M. (1998) Free ricin A chain, proricin, and native toxin have different cellular fates when expressed in tobacco protoplasts. J Biol Chem, ITi, 14194-14199. 

Rosenberg N, Reichman M, Gera A, et al. Antiviral activity of natural and recombinant human leukocyte interferons in tobacco protoplasts. Virology, 1985 140(1) 173-178. 

Mori, M., et ah, Efficient production of human gamma interferon in tobacco protoplasts by genetically engineered brome mosaic vims RNAs. J Gen Virol, 1993 74 (Pt 7) 1255-1260. 

More recently, auxin effects on ion transport have been characterized in protoplasts from Vida faba guard cells and tobacco protoplasts. In both systems, patch clamp techniques have been used to demonstrate auxin regulation of anion channels [14,15]. Alternatively, electrode impalement of protoplasts from faba guard cells has demonstrated that low concentrations of auxin promote inward movement of K, while higher concentrations inhibit this same activity. These effects were correlated with opening and closing of the stomata, respectively [16]. 

Use of Gus in plant protoplasts. Some enzyme mixtures (especially rhoz3mie) used to prepare protoplasts have a very large amount of beta-glucuronidase activity. The usual flotation and KCl washes work well to eliminate all residual activity from tobacco protoplasts, but other types of cells may be more or less difficult to clean up. When transient expression studies or any other protoplast work is performed, it is therefore essential to use the appropriate controls. 

Electroporation-Mediated DNA Transfer to Tobacco Protoplasts for Transient Gene Expression Assays... 

Protoplasts of other plant species can be electroporated with similar procedures as described here for tobacco protoplasts. Pulse length and field strength should, however, be optimized for each species and genotype. Optimal DNA uptake usually requires conditions that are lethal for part of the protoplasts. Likewise, the optimal time point at which to measure transient expression has to be determined empirically. 

Thus, molecular tools (antibodies and cDNA probes) to investigate auxin action are now available. We are now in the process of utilizing the polyclonal and monoclonal antibodies for immunopurification and also to probe receptor structure and function in maize and in other species. For example, preliminary results from Prof. J. Guern s laboratory (by H. Barbier-Brygoo, Gif) show that the antiserum blocks auxin-induced hyperpolarization of tobacco protoplasts. We are also seeking to use the antibodies to examine receptor features in auxin response mutants of several species and to explore signal transduction pathways. 

The rates of incorporation by isolated tobacco protoplasts of thymidine, uridine and leucine were reversibly lowered by coumarin, which consistently inhibited cell... 

Cooper, J. B., Heuser, J. E., and Varner, J. E., 1994, 3,4-dehydroproline inhibits cell wall assembly and cell division in tobacco protoplasts. Plant Physiol. 104 747-752. 

Kato N, Esaka M. Expansion of transgenic tobacco protoplasts expressing pumpkin ascorbate oxidase is more rapid than that of wild-type protoplasts. Planta 2000 210 1018-1022. 

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