Abstract
T-DNA activation tagging was used to generate tobacco mutants with increased tolerance to antimicrotubular herbicides and chilling stress. After transformation, protoplast-derived calli were screened for tolerance to treatments that affect microtubule assembly. In one screen mutants with tolerance to aryl carbamates (a blocker of microtubule assembly) were selected, the second screen was targeted to chilling-tolerant mutants that could survive for several months at 3°C, a third screen combined both factors. The resistance of these mutants to aryl carbamates or chilling was accompanied by resistance of microtubules to these factors. The carbamate tolerant mutants were cross-resistant to chilling stress. This was mirrored by an adaptive reorganization of microtubules and a reduction of microtubule dynamics in response to chilling. The analysis of these mutants suggests (1) that microtubule dynamics limit the tolerance to chilling and EPC, and (2) that the cold sensitivity of microtubules limits chilling tolerance in tobacco.
Similar content being viewed by others
References
Anthony RG and Hussey PJ (1998) Suppression of endogenous α-and β-tubulin synthesis in transgenic maize calli overexpressing α-and β-tubulins. Plant J 16: 297–304.
Anthony RG, Reichelt S and Hussey PJ (1999) Dinitroaniline herbicide-resistant transgenic tobacco plants generated by cooverexpression of a mutant α-tubulin and a β-tubulin. Nature Biotechnol 17: 712–716.
Baird V, Blume YB and Wick SM (2000) Microtubular and cytoskeletal mutants. In: Nick P (ed.), Plant Microtubules: Potential for Biotechnology. (pp. 159–191) Springer, Berlin, Heidelberg.
Bartolo ME and Carter JV (1991) Effect of microtubule stabilization on the freezing tolerance of mesophyll cells of spinach. Plant Physiol 97: 182–187.
Bartolo ME and Carter JV (1992) Lithium decreases cold-induced microtubule depolymerization in mesophyll cells of spinach. Plant Physiol 99: 1716–1718.
Bin-Bing Zh and Kirschner MW (1999) Quantitative measurement of the catastrophe rate of dynamic microtubules. Cell Mot Cytoskeleton 43: 43–51.
Breviario D and Nick P (2000) Plant tubulins: a melting pot for basic questions and promising applications. Transgenic Res 9: 383–393.
Caboche M (1980) Nutritional requirements of protoplasts derived haploid tobacco cells grown at low densities in liquid medium. Planta 149: 1–18.
Caudron N, Valiron O, Usson Y, Valiron P and Job D (2000) A reassessment of the factors affecting microtubule assembly and disassembly in vitro. J Mol Biol 297: 211–220.
Chang C, Kwok SF, Bleecker AB and Meyerowitz EM (1993) Arabidopsis ethylene-response gene ETR1: similarity of product to two-component regulators. Science 262: 539–544.
Chu B, Snustad DP and Carter JV (1993) Alteration of beta-tubulin gene expression during low temperature exposure in leaves of Arabidopsis thaliana. Plant Physiol 103: 371–377.
Cleveland DW (1988) Autoregulated instability of tubulin mRNAs: a novel eukaryotic regulatory mechanism. Trends Biochem Sci 13: 339–343.
Ding JP and Pickard BG (1993) Mechanosensory calcium-selective cation channels in epidermal cells. Plant J 3: 83–110.
Durso NA and Cyr RJ (1994) A calmodulin-sensitive interaction between microtubules and a higher plant homolog of elongation factor 1α. Plant Cell 6: 893–905.
Feldmann KA (1991) T-DNA insertional mutagenesis in Arabidopsis: mutational spectrum. Plant J 1: 71–82.
Fisher DD, Gilory S and Cyr RJ (1996) Evidence for opposing effects of calmodulin on cortical microtubules. Plant Physiol 122: 1070–1087.
Jian LC, Sun LH and Liu ZP (1989) Studies on microtubule cold stability in relation to plant cold hardiness. Acta Bot Sinica 31: 737–741.
Hayashi H, Czaja I, Schell J and Walden R (1992) Activation of plant gene by T-DNA tagging: auxin independent growth in vitro. Science 258: 1350–1353.
Kakimoto T (1996) CKI1, a histidine kinase homolog implicated in cytokinin signal transduction. Science 274: 982–985.
Kao RN and Michayluk MR (1975) Nutritional requirements for growth of Vicia hajastana cells at very low population density in liquid medium. Planta 126: 105–110.
Kerr GP and Carter JV (1990) Relationship between freezing tolerance of root-tip cells and cold stability of microtubules in rye (Secale cereale L. cv Puma). Plant Physiol 93: 77–82.
Kioka N, Sakata S, Kawauchi T, Amachi T, Akiyama SK, Okazaki K et al. (1999) Vinexin: a novel vinculin-binding protein with multiple SH3 domains enhances actin cytoskeletal organization. J Cell Biol 144: 59–69.
Knight MR, Campbell A, Smith SM and Trewavas AJ (1991) Transgenic plant aequorin reports the effects of touch and cold shock and elicitors on cytoplasmic calcium. Nature 352: 524–526.
Koncz C, Martini N, Mayerhofer R, Koncz-Kalman Z, Körber H, Redei GP et al. (1989) High frequency T-DNA mediated gene tagging in plants. Proc Natl Acad Sci USA 86: 8467–8471.
Koncz C, Mayerhofer R, Koncz-Kalman Z, Nawrath C, Reiss B, Redei GP et al. (1990) Isolation of a gene encoding a novel chloroplast protein by T-DNA tagging in Arabidopsis thaliana. EMBO J 9: 1337–1346.
Koncz C, Martini N, Szabados L, Hrouda M, Bachmair A and Schell J (1994) Specialized vectors for gene tagging and expression studies. In: Gelvin SB and Schilperoort RA (eds), Plant Molecular Biology Manual B2. (pp. 1–22) Kluwer Academic Publishers, Belgium.
Kreis TE (1987) Microtubules containing detyrosinylated tubulin are less dynamic. EMBO J 6: 2597–2606.
Kumagai H and Nishida E (1979) The interaction between calcium dependent regulator protein of cyclic nucleotide phosphodiesterase and microtubule proteins II, association of calcium-dependent regulator protein with tubulin dimers. J Biochem 85: 1267–1274.
Maliga P, Sz-Breznovits A and Marton L (1973) Streptomycinresistant plants from callus culture of haploid tobacco. Nat New Biol 244: 29–30.
Mazars C, Thion L, Thuleau P, Graziana A, Knight MR, Moreau M et al. (1997) Organization of cytoskeleton controls the changes in cytosolic calcium of cold-shocked Nicotiana plumbaginifolia protoplasts. Cell Calcium 22: 413–420.
Mizuno K (1992) Induction of cold stability of microtubules in culture tobacco cells. Plant Physiol 100: 740–748.
Mizuno K and Suzaki T (1991) Effects of anti-microtubule drugs on in vitro polymerization of tubulin from mung mean. Bot Mag Tokyo 103: 435–448.
Monroy AF, Sarhan F and Dhindsa RS (1993) Cold-induced changes in freezing tolerance, protein phosphorylation, and gene expression. Plant Physiol 102: 1227–1235.
Murashige T and Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol Plantarum 15: 473–497.
Nick P (2000) Control of the response to low temperatures. In: Nick P (ed.), Plant Microtubules: Potential for Biotechnology. (pp. 121–135) Springer, Berlin, Heidelberg.
Nick P, Yatou O, Furuya M and Lambert AM (1994) Auxindependent microtubule response and seedling development are affected in a rice mutant resistant to EPC. Plant J 6: 651–663.
Nick P, Heuing A and Ehmann B (2000) Plant chaperonins: a role in microtubule-dependent wall formation. Protoplasma 211: 234–244.
Potrykus I and Shilito RD (1986) Protoplasts: isolation, culture, plant regeneration. Plant Mol Biol 118: 549–578.
Putnam-Evans C, Harmon AC, Palevitz BA, Fechheimer M and Cormier MJ (1989) Calcium-dependent protein kinase is localized with F-actin in plant cells. Cell Motil Cytoskel 12: 12–22.
Rikin A, Atsmon D and Gitler C (1980) Chilling injury in cotton (Gossypium hirsutum L.): Effect of antimicrotubular drugs. Plant Cell Physiol 21: 829–837.
Schell J, Bisseling T, Dülz M, Frassen H, Fritze K, John M et al. (1999) Re-evaluation of phytohormone-independent division of tobacco protoplasts-derived cells. Plant J 17(5): 461–466.
Thion L, Mazars C, Thuleau P, Graziana A, Rossignol M, Moreau M et al. (1996) Activation of plasma membrane voltage-dependent calcium-permeable channels by disruption of microtubules in carrot cells. FEBS Lett 393: 13–18.
Thomashow MF (2001) So what's new in the field of plant cold acclimation? Lots! Plant Physiol 125: 89–93.
Vaughn KC (2000) Anticytoskeletal herbicides. In: Nick P (ed.), Plant Microtubules: Potential for Biotechnology. (pp. 193–205) Springer, Berlin, Heidelberg.
Walden R, Fritze K and Harling H (1995) Induction of signal transduction pathways through promoter activation. Meth Cell Biol 49: 455–469.
Watson DJ (1952) The physiological basis of variation yield. Adv Agron 4: 101–145.
Weigel D, Ahn JH, Bla'zquez MA, Borevitz JO, Christensen SK, Frankhauser C et al. (2000) Activation tagging in Arabidopsis. Plant Physiol 22: 1003–1013.
Wiesler B, Wang QY and Nick P (2002) The stability of cortical microtubules depends on their orientation Plant J 32(6): 1023–1032.
Zubko E, Adams CJ, Machaé'kova'I, Malbeck J, Scollan C and Meyer P (2002) Activation tagging identifies a gene from Petunia hybrida responsible for the production of active cytokinins in plants Plant J 29(6): 797–808.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ahad, A., Wolf, J. & Nick, P. Activation-Tagged Tobacco Mutants that are Tolerant to Antimicrotubular Herbicides are Cross-Resistant to Chilling Stress. Transgenic Res 12, 615–629 (2003). https://doi.org/10.1023/A:1025814814823
Issue Date:
DOI: https://doi.org/10.1023/A:1025814814823