Abstract
The role of ethylene in vegetative bud formation was investigated using transgenic tobacco plants expressing an antisense tomato 1-aminocyclopropane-carboxylic acid synthase (ACS) gene. Northern blot hybridization showed that the accumulation of ACS mRNA was strongly reduced in the bud-forming leaf explants of the transgenic plants. Consequently, these transgenic tissues exhibited low ACS enzyme activity, 1-aminocyclopropane-carboxylic acid (ACC) content and ethylene production, and at the same time the tissue capacity to generate buds was greatly enhanced. However, it was also noted that the antisense ACS gene did not inhibit the endogenous ACS gene expression in intact transgenic tobacco plants. The growth and development of the transgenic tobacco was almost identical to control plants with respect to height, internode number, leaf morphology, and flowering time. Furthermore, mature leaves of transgenic tobacco had similar chlorophyll content, stomatal conductance, photosynthetic ability, and transpiration rates compared to control plants. These results demonstrated that ethylene plays an important role in bud formation in tobacco tissue culture.
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Alexander L, Grierson D. 2002. Ethylene biosynthesis and action in tomato: a model for climacteric fruit ripening. J Exp Bot 53:2039–2055
Amor MB, Guis M, Latche A, Bouzayen M, Pech JC, Roustan JP. 1998. Expression of an antisense 1-aminocyclopropane-1-carboxylate oxidase gene stimulates shoot regeneration in Cucumis melo. Plant Cell Rep 17:586–589
Biondi S, Scaramagli S, Capitani F, Marino G, Altamura MM, Torrigiani P. 1998. Ethylene involvement in vegetative bud formation in tobacco thin layers. Protoplasma 202:134–144
Bleecker AB, Kende H. 2000. Ethylene: a gaseous signal molecule in plants. Annu Rev Cell Dev Biol 16:1–18
Bradford MM 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Capell T, Bassie L, Topsom L, Hitchin E, Christou P. 2000. Simultaneous reduction of the activity of two related enzymes, involved in early steps of the polyamine biosynthetic pathway, by a single antisense cDNA in transgenic rice. Mol Gen Genet 264:470–476
Dellaporta SL, Wood J, Hicks JB. 1983. A plant DNA minipreparation: versionII. Plant Mol Biol Rep 1:19–21
Frohman MA, Dush MK, Martin GR. 1988. Rapid amplification of full-length cDNA ends from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci USA 85:8998–9002
González A, Arigita L, Majada J, Tamés RS. 1997. Ethylene involvement in in vitro organogenesis and plant growth of Populus tremula L. Plant Growth Regul 22:1–6
Hamilton AJ, Lycett JW, Grierson D. 1990. Antisense gene that inhibits synthesis of the hormone ethylene in transgenic plants. Nature 346:284–287
Kende H 1993. Ethylene biosynthesis. Annu Rev Plant Physiol Plant Mol Biol 44:283–307
Kumar PP, Lakshmanan P, Thorpe TA. 1998. Regulation of morphogenesis in plant tissue culture by ethylene. In Vitro Cell Dev Biol Plant 34:94–103
Ma QH, Song YR. 1997. Expression of tomato antisense ACC synthase gene in transgenic tobacco and its role in shoot formation. Acta Bot Sinica 39:1047–1052
Martinis DD, Mariani C. 1999. Silencing gene expression of the ethylene-forming enzyme results in a reversible inhibition of ovule development in transgenic tobacco plants. Plant Cell 11:1061–1071
Mol JNM, Van der Krol AR, Van Tunen R, Van Blokland R, Lange PD, Stuitje AR. 1990. Regulation of plant gene expression by antisense RNA. FEBS Lett 268:427–430
Nara A, Takeuchi Y. 2002. Ethylene evolution from tobacco leaves irradiated with UV-B. J Plant Res 115:247–253
Oeller PW, Li MW, Taylor LP, Pike DA, Theologis A. 1991. Reversible inhibition of tomato fruit senescence by antisense RNA. Science 254:437–439
Porra RL, Thomson WA, Kriedemann PE. 1989. Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochim Biophys Acta 975:382–394
Pua EC, Deng X, Koh ATC. 1999. Genotypic variability of de novo shoot morphogenesis of Brassica oleracea in vitro in response to ethylene inhibitors and putrescine. J Plant Physiol 155:598–605
Pua EC, Lee JEE. 1995. Enhance de novo shoot morphogenesis in vitro by expression of antisense 1-aminocyclopropane-1-carboxylase oxidase gene in transgenic mustard plants. Planta 196:69–76
Sambrook J, Fritsch EF, Maniatis T. 1989. Molecular Cloning: a Laboratory Manual, 2nd ed, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA
Scaramagli S, Biondi S, Capitani F, Gerola P, Altamura MM, Torrigiani P. 1999. Polyamine conjugate levels and ethylene biosynthesis: inverse relationship with vegetative bud formation in tobacco thin layers. Physiol Plant 105:367–376
Temple SJ, Knight TJ, Unkefer PJ, Sengupta-Gopalan C. 1993. Modulation of glutamine synthetase gene expression in tobacco by the introduction of an alfalfa glutamine synthetase gene in sense and antisense orientation: molecular and biochemical analysis. Mol Gen Genet 236:315–325
Torrigiani P, Scaramagli S, Castiglione S, Altamura MM, Biondi S. 2003. Downregulation of ethylene production and biosynthetic gene expression is associated to changes in putrescine metabolism in shoot forming tobacco thin layers. Plant Sci 164:1087–1094
Torrigiani P, Scaramagli S, Ziosi V, Mayer M, Biondi S. 2005. Expression of an antisense Datura stramonium S-adenosylmethionine decarboxylase cDNA in tobacco: changes in enzyme activity, putrescine-spermidine ratio, rhizogenic potential, and response to methyl jasmonate. J Plant Physiol 162:559–571
Trung-Nghia P, Bassie L, Safwat G, Thu-Hang P, Lepri O, others. 2003. Reduction in the endogenous arginine decarboxylase transcript levels in rice leads to depletion of the putrescine and spermidine pools with no concomitant changes in the expression of downstream genes in the polyamine biosynthetic pathway. Planta 218:125–134
Acknowledgments
This work was supported by grants from the National Natural Science Foundation of China (No. 30400222 and No.30570133), the Natural Science Foundation of Beijing (No.503211), the Chinese National Special Foundation for Transgenic Plant Research and Commercialization (J2002-B007), and the Innovation Project of Chinese Academy of Sciences. The authors thank Prof A Theologis (the Plant Gene Expression Center, Berkeley, CA, USA) for providing tomato ACS cDNA. We also sincerely thank Dr. Charles H. Hocart (Australian National University, Australia) for critical reading of the manuscript.
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Ma, QH., Li, YL. Inhibition of Ethylene Biosynthesis Enhances Vegetative Bud Formation without Affecting Growth and Development of Transgenic Tobacco Plants. J Plant Growth Regul 25, 101–109 (2006). https://doi.org/10.1007/s00344-005-0086-y
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DOI: https://doi.org/10.1007/s00344-005-0086-y