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Expression of the Arabidopsis AtAux2-11 auxin-responsive gene in transgenic plants

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Abstract

Five constructions containing deletions of the promoter from an auxin-inducible gene of Arabidopsis thaliana, AtAux2-11, were fused to the coding region of the reporter gene LacZ, which encodes β-galactosidase, and a polyadenylation 3′-untranslated nopaline synthase sequence from Agrobacterium. These chimeric genes were introduced into Arabidopsis by Agrobacterium tumefaciens-mediated transformation, and expression of the gene was examined by spectrophotometric and histochemical analyses. A 600 bp fragment from the AtAux2-11 promoter conferred histochemical patterns of staining similar to the longest 5′ promoter tested, a 3.0 kb fragment. Localization of AtAux2-11/LacZ activity in the transgenic plants revealed spatial and temporal expression patterns that correlated with tissues and cells undergoing physiological processes modulated by auxin. LacZ activity was expressed in the elongating region of roots, etiolated hypocotyls, and anther filaments. Expression was detected in the vascular cylinder of the root and the vascular tissue, epidermis, and cortex of the hypocotyl, and filament. The AtAux2-11/LacZ gene was preferentially expressed in cells on the elongating side of hypocotyls undergoing gravitropic curvature. Expression of the chimeric gene in the hypocotyls of light-grown seedlings was less than that in etiolated seedling hypcotyls. The AtAux2-11/LacZ gene was active in the root cap, and expression in the root stele increased at sites of lateral root initiation. Staining was evident in cell types that develop lignified cell walls, e.g. trichomes, anther endothecial cells, and especially developing xylem. The chimeric gene was not expressed in primary meristems. While the magnitude of expression increased after application of exogenous auxin (2,4-D), the histochemical localization of AtAux2-11/LacZ remained unchanged.

Transgenic plants with a 600 bp promoter construct (−0.6 kb AtAux2-11/LacZ) had higher levels of basal and auxin-inducible expression than plants with a 3.0 kb promoter construct. Transgenic plants with a −500 bp promoter had levels of expression similar to the −3.0 kb construct. The −0.6 kb AtAux2-11/LacZ gene responded maximally to a concentration of 5 × 10−6 to 5 × 10−5 M 2,4-D and was responsive to as little as 5 × 10−8 M. The evidence presented here suggests that this gene may play a role in several auxin-mediated developmental and physiological processes.

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References

  1. Ainley WM, Walker JC, Nagao RT, Key JL: Sequence and characterization of two auxin-regulated genes from soybean. J Biol Chem 263: 10658–10666 (1988).

    PubMed  Google Scholar 

  2. Alliote T, Tiré C, Engler G, Peleman J, Caplan A, VanMontagu M, Inzé D: An auxin-regulated gene of Arabidopsis thaliana encodes a DNA-binding protein. Plant Physiol 89: 743–752 (1989).

    Google Scholar 

  3. An G, Mitra A, Choi H, Costa M, An K, Thornburg RW, Ryan C: Functional analyses of the 3′ control region of the potato wound-inducible protease inhibitor II gene. Plant Cell 1: 115–122 (1989).

    Article  PubMed  Google Scholar 

  4. Baulcombe DC, Key JL: Polyadenylated RNA sequences which are reduced in concentration following auxin treatment of soybean hypocotyls. J Biol Chem 255: 8907–8913 (1980).

    Google Scholar 

  5. Blakely LM, Blakely RM, Colowit PM, Elliot DS: Experimental studies on lateral root formation in radish seedling roots. II. Analysis of the dose-response to exogenous auxin. Plant Physiol 87: 414–419 (1988).

    Google Scholar 

  6. Conner TW, Goekjian VH, Lafayette PR, Key JL: Structure and expression of two auxin-inducible genes from Arabidopsis. Plant Mol Biol 15: 623–632 (1990).

    PubMed  Google Scholar 

  7. Ditta G, Stanfield G, Corbin D, Helinski DR: Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci USA 77: 7347–7354 (1980).

    PubMed  Google Scholar 

  8. Figurski DH, Helinski DR: Replication of an origin containing a derivative of plasmid RK2, dependence on a plasmid function provided in trans. Proc Natl Acad Sci USA 76: 1648–1652 (1979).

    PubMed  Google Scholar 

  9. Gee MA, Hagen G, Guilfoyle TJ: Tissue-specific and organ-specific expression of soybean auxin-responsive transcripts GH3 and SAURs. Plant Cell 3: 419–430 (1991).

    Article  PubMed  Google Scholar 

  10. Guilfoyle TJ: Auxin-regulated gene expression in higher plants. CRC Crit Rev Plant Sci 4: 247–276 (1986).

    Google Scholar 

  11. Hagen G, Kleinschmidt A, Guilfolye TJ: Auxin-regulated gene expression in intact soybean hypocotyl and excised hypocotyl sections. Planta 162: 147–153 (1984).

    Google Scholar 

  12. Hagen G, Guilfoyle TJ: Rapid induction of selective transcription by auxins. Mol Cell Biol 5: 1197–1203 (1985).

    PubMed  Google Scholar 

  13. Hood EE, Helmer GL, Fraley RT, Chilton M-D: The hypervirulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of the T-DNA. J Bact 168: 1291–1298 (1986).

    PubMed  Google Scholar 

  14. Jeffs RA, Northcote DH: The influence of indolyl-3-acetic acid and sugar on the pattern of induced differentiation in plant tissue culture. J Cell Sci 2: 77–88 (1967).

    PubMed  Google Scholar 

  15. Key J, Kroner P, Walker J, Hong JC, Ulrich TH, Ainley WM, Gantt JS, Nagao RT: Auxin-regulated gene expression. Phil Trans R Soc Lond B 314: 427–440 (1986).

    Google Scholar 

  16. Laemmli UK: Cleavage of structural proteins during the assembly of bacteriophage T4. Nature 277: 680–685 (1970).

    Google Scholar 

  17. Li Y, Hagen G, Guilfoyle TJ: An auxin-responsive promoter is differentially induced by auxin gradients during tropisms. Plant Cell 3: 1167–1175 (1991).

    Article  PubMed  Google Scholar 

  18. Lyndon RF: Plant Development: The Cellular Basis. Unwin-Hyman, London (1990).

    Google Scholar 

  19. McClure BA, Guilfoyle T: Characterization of a class of small auxin-inducible soybean polyadenylated RNAs. Plant Mol Biol 9: 611–623 (1987).

    Google Scholar 

  20. Miller JH. Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1972).

    Google Scholar 

  21. Reddy ASN, Poovaiah BW: Molecular cloning and sequencing of a cDNA for an auxin-repressed mRNA: correlation between fruit growth and repression of the auxinregulated gene. Plant Mol Biol 14: 127–136 (1990).

    PubMed  Google Scholar 

  22. Sachs T: The control of patterned differentiation of vascular tissues. Adv Bot Res 9: 151–262 (1981).

    Google Scholar 

  23. Sachs T: Axiality and polarity in plants. In: Barlow PW, Carr DJ (eds) Positional Controls in Plant Development, pp. 193–224. Cambridge University Press, Cambridge (1984).

    Google Scholar 

  24. Takahashi Y, Kuroda H, Tanaka T, Machida Y, Takabe I, Nagata T: Isolation of an auxin-regulated cDNA expressed during the transition from G0 to S phase in tobacco mesophyll protoplast. Proc Natl Acad Sci USA 86: 9279–9283 (1989).

    PubMed  Google Scholar 

  25. Teeri TH, Lehväslaiho H, Franck M, Uotila J, Heino P, Palva ET, VanMontagu M, Herrera-Estrella L: Gene fusions to lacZ reveal new expression patterns of chimeric genes in transgenic plants. EMBO J 8: 343–350 (1989).

    PubMed  Google Scholar 

  26. Theologis A, Huynh TV, Davis RW: Rapid induction of specific mRNAs by auxin in pea epicotyl tissue. J Mol Biol 3: 53–68 (1985).

    Google Scholar 

  27. Valvekens D, VanMontagu M, VanLijsebettens M: Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc Natl Acad Sci USA 85: 5536–5540 (1980).

    Google Scholar 

  28. van derZaal EJ, Mennes AM, Libbenga KR. Auxin-induced rapid changes in translatable mRNAs in tobacco cell suspensions. Planta 172: 514–519 (1987).

    Google Scholar 

  29. Walker JC, Key JL: Isolation of cloned cDNAs to auxinresponsive poly(A)+ RNAs of elongating soybean hypocotyl. Proc Natl Acad Sci USA 79: 7185–7189 (1982).

    Google Scholar 

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Author notes

  1. Robert E. Wyatt

    Present address: Department of Math and Science, University of the Virgin Islands, USVI 00802, St. Thomas, USA

  2. W. Michael Ainley

    Present address: Dow Elanco Inc., 46206, Indianapolis, IN, USA

  3. Timothy W. Conner

    Present address: Monsanto Co., 63198, St. Louis, MO, USA

Authors and Affiliations

  1. Department of Botany, University of Georgia, 30602, Athens, GA, USA

    Robert E. Wyatt, W. Michael Ainley, Ron T. Nagao, Timothy W. Conner & Joe L. Key

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  1. Robert E. Wyatt
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  2. W. Michael Ainley
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  3. Ron T. Nagao
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  4. Timothy W. Conner
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  5. Joe L. Key
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Wyatt, R.E., Ainley, W.M., Nagao, R.T. et al. Expression of the Arabidopsis AtAux2-11 auxin-responsive gene in transgenic plants. Plant Mol Biol 22, 731–749 (1993). https://doi.org/10.1007/BF00027361

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  • DOI: https://doi.org/10.1007/BF00027361

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