Summary
The iaaL gene of Pseudomonas syringae subsp. savastanoi encodes an indoleacetic acid-lysine synthetase that conjugates lysine to indoleacetic acid. A chimaeric gene consisting of the iaaL coding region under the control of the 35S RNA promoter from cauliflower mosaic virus (35SiaaL) has been used to test if iaaL gene expression leads to morphological alterations in tobacco and potato. Transgenic tobacco plantlets bearing this construct have been shown to synthesize IAA-[14C]lysine when fed with [14C]lysine. In late stages of development, their leaves show an increased nastic curvature (epinasty) of the petiole and midvein, a finding suggestive of an abnormal auxin metabolism. The alteration is transmitted to progeny as a dominant Mendelian trait cosegregating with the kanamycin resistance marker. Transgenic potato plants harbouring the construct are also characterised by petiole epinasty. Moreover, 35SiaaL transgenic plants have an increased internode length in potato and decreased root growth in both tobacco and potato. An increased content of IAA-conjugates in leaf blade was found to correlate with the epinastic alterations caused by iaaL gene expression in tobacco leaves. These data provide evidence that IAA conjugation is able to modulate hormone action, suggesting that the widespread endogenous auxin-conjugating activities are of physiological importance.
Similar content being viewed by others
References
Bandurski RS, Schulze A (1977) Concentration of indole-3-acetic acid and its derivatives in plants. Plant Physiol 60:211–213
Bearder JR (1980) Plant hormones and other growth substances — Their background, structures and occurrence. In: MacMillan J (ed) Encyclopedia of Plant Physiology, vol 9, 9–80. Springer, Berlin Heidelberg New York
Benfey PN, Chua NH (1989) Regulated genes in transgenic plants. Science 244:174–181
Bialek K, Cohen JD (1989) Quantitation of indoleacetic acid conjugates in bean seeds by direct tissue hydrolysis. Plant Physiol 90:398–400
Bialek K, Meudt NJ, Cohen JD (1983) Indole-3-acetic acid (IAA) and IAA conjugates applied to bean stem sections. Plant Physiol 73:130–134
Boylan MT, Quail PT (1989) Oat phytocrome is biologically active in transgenic tomatoes. Plant Cell 1: 765–773
Burström HG (1969) Influence of the tonic effect of gravitation and auxin on cell elongation and polarity in roots. Amer J Bot 56:679–684
Cardarelli M, Mariotti D, Pompeni M, Spano L, Capone I, Costantino P (1987) Agrobacterium rhizogenes T-DNA genes capable of inducing hairy root phenotype. Mol Gen Genet 209:475–480
Cohen JD (1982) Identification and quantitative analysis of indole3-acetyl-L-aspartate from seeds to Glycine max L. Plant Physiol 70:749–753
Cohen JD, Bandurski RS (1982) Chemistry and physiology of the bound auxins. Annu Rev Plant Physiol 33:403–430
Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: Version II. Plant Mol Rep 1:19–21
Evans ML (1984) Functions of hormones at the cellular level of organization. In: Scott TK (ed) Encyclopedia of Plant Physiology. Springer, Berlin Heidelberg New York Tokyo, vol 10, pp 23–62
Glass NL, Kosuge T (1986) Cloning of the gene for indoleacetic acid-lysine synthetase from Pseudomonas syringae subsp. savastanoi. J Bacteriol 166:598–603
Hamilton AJ, Lycett GW, Grierson D (1990) Antisense gene that inhibits synthesis of the hormone ethylene in transgenic plants. Nature 346:284–287
Hangartner RP, Peterson MD, Good NE (1980) Biological activities of indoleacetyl amino acids and their use as auxins in tissue culture. Plant Physiol 65:761–767
Horsch R, Fraley R, Rogers S, Sanders P, Lloyd A, Hoffmann W (1984) Inheritance of functional foreign genes in plants. Science 223:496–498
Hutzinger O, Kosuge T (1968) Microbial synthesis and degradation of indole-3-acetic acid. III. The isolation and characterization of indole-3-acetyl-ɛ-l-lysine. Biochemistry 7:601–605
Jackson MB (1985) Ethylene and responses of plants to soil waterlogging and submergence. Annu Rev Plant Physiol 36:145–174
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907
Jouanin L, Vilaine F, Tourneur J, Tourneur C, Pautot V, Muller JF, Caboche M (1987) Transfer of a 4.3 kb fragment of the T l -DNA of Agrobacterium rhizogenes strain A4. Mol Gen Genet 206:387–392
Keller MJ, Shanklin J, Vierstra RD, Hershey HP (1989) Expression of a functional monocotyledonous phytochrome in transgenic tobacco. EMBO J 8:1005–1012
Klee HJ, Horsch RB, Hinchee MA, Hein MB, Hoffmann NL (1987) The effects of overproduction of two Agrobacterium tumefaciens T-DNA auxin biosynthetic gene products in transgenic petunia plants. Genes Dev 1: 86–96
Knapp S, Coupland G, Uhrig H, Starlinger P, Salamini F (1988) Transposition of the maize transposable element Ac in Solanum tuberosum. Mol Gen Genet 204:383–386
Koncz C, Schell J (1986) The promoter of TL-DNA gene 5 controls the tissue-specific expression of chimaeric genes carried by a novel type of Agrobacterium binary vector. Mol Gen Genet 204:383–396
Kosuge T, Comai L, Glass NL (1983) Virulence determinants in plant-pathogen interactions. In: Goldberg B (ed) Plant Molecular Biology. Alan R Liss, Inc, New York, pp 167–177
Lantican BP, Muir RM (1969) Auxin physiology of dwarfism in Pisum sativum. Physiol Plant 22:412–423
Logemann J, Schell J, Willmitzer L (1987) Improved method for the isolation of RNA from plant tissue. Anal Biochem 163:16–20
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Matthysse AG, Scott TK (1984) Functions of hormones at the whole plant level of organization. In: Scott TK (ed) Encyclopedia of plant physiology, vol 10, 219–243. Springer, Berlin Heidelberg New York
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–496
Nagy JI, Maliga P (1976) Callus induction and plant regeneration from mesophyll protoplasts of Nicotiana sylvestris. Z Pflanzenphysiol 78:453–455
Oono Y, Handa T, Kanaya K, Uchimiya H (1987) the TL-DNA gene of Ri plasmids responsible for dwarfness of tobacco plants. Jpn J Genet 62:501–505
Palmer JH (1985) Epinasty, hyponasty and related topics. In: Pharis RP, Reid DM (eds) Encyclopedia of plant physiology Springer, Berlin Heidelberg New York Tokyo, vol 11, pp 139–168
Pietrzak M, Shillito RD, Hohn T, Potrykus I (1986) Expression in plants of two bacterial antibiotic resistance genes after protoplast transformation with a new plant vector. Nucleic Acids Res 14:5857–5868
Reinecke DM, Bandurski RS (1987) Auxin biosynthesis and metabolism. In: Davies PJ (ed) Plant hormones and their role in plant growth and development. Martinus Nijhoff, Dordrecht Boston Lancaster, pp 24–42
Roberto FF, Klee H, White F, Nordeen R, Kosuge T (1990) Expression and fine structure of the gene encoding indole-3-acetyl-L-lysine synthetase from Pseudomonas savastanoi. Proc Natl Acad Sci USA 87:5797–5801
Roberts JA, Hooley R (1988) In: Plant growth regulators pp 119–121. Blackie and Son Ltd, Glasgow and London
Schmülling T, Schell J, Spena A (1988) Single genes from Agrobacterium rhizogenes influence plant development. EMBO J 7:2621–2629
Simon R, O'Connel M, Labes M, Pühler A (1986) Plasmid vectors for the genetic analysis and manipulation of rhizobia and other gram-negative bacteria. Meth Enzymol 118:640–659
Sinkar VP, Pythoud F, White FF, Nester EW, Gordon MP (1988) RoIA locus of the Ri plasmid directs developmental abnormalities in transgenic tobacco plants. Genes Dev 2:688–697
Spena A, Schmülling T, Koncz C, Schell J (1987) Independent and synergistic activity of rolA, B and C loci in stimulating abnormal growth in plants. EMBO J 6:3891–3899
Spena A, Aalen RA, Schulze SC (1989) Cell-autonomous behavior of the roIC gene of Agrobacterium rhizogenes during leaf development: A visual assay for transposon excision in transgenic plants. Plant Cell 1:1157–1164
Steeves TA, Sussex IM (1989) Patterns in plant development. Cambridge University Press, Cambridge New York New Rochelle Melbourne Sydney
Van Onckelen H, Prinsen E, Chriqui D, Tepfer M, De Greef J (1988) Endogenous auxin levels in normal and hairy root tobacco plants. Med Fac Landbouww Rijkuniv Gent 53:1685–1693
Yates F (1934) Contingency tables involving small numbers and the chi2 test. Supp Jour Roy Stat Soc i:217–235
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Spena, A., Prinsen, E., Fladung, M. et al. The indoleacetic acid-lysine synthetase gene of Pseudomonas syringae subsp. savastanoi induces developmental alterations in transgenic tobacco and potato plants. Molec. Gen. Genet. 227, 205–212 (1991). https://doi.org/10.1007/BF00259672
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00259672