Skip to main content
Log in

Potential use of theaux2 gene fromAgrobacterium rhizogenes as a conditional negative marker in transgenic cabbage

  • Published:
Transgenic Research Aims and scope Submit manuscript

Abstract

An originalAgrobacterium tumefaciens-mediated transformation procedure, based on the actions of both wild type and disarmed bacterial strains, was developed. Theaux2 gene ofA. rhizogenes was introduced into a rapid-cycling genotype of cabbage (Brassica oleracea L.). Theaux2 gene product converts naphthalene acetamide into the auxin naphthalene acetic acid. Expression of this gene in the transgenic progeny grownin vitro led to an altered root phenotype. On a medium supplemented with napthalene acetamide (NAM), two of the three analysed progenies were characterized by the formation of callus instead of roots, whereas on a NAM-free medium all the plantlets from these progenies presented a normal phenotype. Expression of theaux2 gene was also assessed under horticultural conditions by sowing seeds in sand and watering them with a nutritive solution supplemented with NAM. Under these conditions, NAM inhibited the formation of a root system in transgenic plantlets and induced the death of the transgenic plantlets three to four weeks after germination. Thus,aux2 acts as a lethal conditional marker which could be used in negative selection of cabbage. Potential utilization of theaux2 gene to screen spontaneous androgenetic plants in order to transfer cytoplasmic male sterility in a single generation is discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bannerot, H. and Boulidard, L. (1974) Transfer of cytoplasmic male sterility fromRaphanus sativus toBrassica oleracea. InProceedings of the Eucarpia Meeting on Cruciferae, Scottish Horticultural Research Institute, Invergourie, Dundee, pp. 52–5.

    Google Scholar 

  • Boyer, H.W. and Rolland-Dussoix, D. (1969) A complementation analysis of the restriction and modification of DNA inEscherichia coli.J. Mol. Biol.,41, 459–72.

    Article  PubMed  Google Scholar 

  • Brasileiro, A. C. M., Leplé, J. C., Muzzin, J., Ounnough, D., Michel, M. F. and Jouanin, L. (1991) An alternative approach for gene transfer in trees using wild-typeAgrobacterium strains.Plant. Mol. Biol.,17, 441–52.

    Article  PubMed  Google Scholar 

  • Braun, A. C. (1948) Studies on the origin and development of plant teratomas incited by the crown gall bacterium.Amer. J. Bot.,35, 511–9.

    Google Scholar 

  • Budar, F., Deboeck, F., Van Montagu, M. and Hernalsteens, J. P. (1986) Introduction and expression of the octopine T-DNA oncogenes in tobacco plants and their progeny.Plant Sci.,46, 195–206.

    Article  Google Scholar 

  • Burk, L. G. (1962) Haploids in genetically marked progenies of tobacco.J. Hered.,53, 222–5.

    Google Scholar 

  • Camilleri, C. and Jouanin, L. (1991) The TR-DNA region carrying the auxin synthesis genes of theAgrobacterium rhizogenes agropine-type plasmid pRiA4: nucleotide sequence analysis and introduction into tobacco plants.Mol. Plant-Microbe Interact.,4, 155–162.

    PubMed  Google Scholar 

  • Chase, S.S. (1963) Androgenesis—its use for transfer of maize cytoplasm.J. Hered. 54, 152–8.

    Google Scholar 

  • Chase, S. S. (1960) Monoploids and monoploid-derivatives of maize (Zea mays L.).Bot. Rev.,35, 117–67.

    Google Scholar 

  • Coïc, Y. and Lesaint, C. (1961) Comment assurer une bonne nutrition en eau et ions minéraux en horticulture.Hortic. Fr.,8, 11–14.

    Google Scholar 

  • Dellaporta, S. L., Wood, J., and Hicks, J. B. (1983) A plant DNA minipreparation: versin II.Plant Mol. Biol. Rep.,1, 19–21.

    Google Scholar 

  • Deroles, S. C. and Gardner, R. C. (1988) Analysis of the T-DNA structure in a large number of transgenic petunias generated byAgrobacterium-mediated transformation.Plant Mol. Biol.,11, 365–77.

    Article  Google Scholar 

  • Doré, C. (1975) La multiplication clonale de l'asperge (Asparagus officinalis) par culturein vitro: son utilisation en sélection.Ann. Amélior. Plantes,25, 201–24.

    Google Scholar 

  • Doré, C. and Boulidard, L. (1988) Production de plantes androgénétiques de chou à choucroute (Brassica oleracea L. ssp.capitata) par culture d'anthèresin vitro: comportement des lignées haploïdes doublées (HD) et leur intérêt comme parents d'hybrides F1.Agronomie,8, 851–62.

    Google Scholar 

  • Hoekema, A., Hirsch, P. R., Hooykaas, P. J. J. and Schilperoot, R. A. (1983) A binary plant vector strategy based on separation of vir- and T-region of theAgrobacterium tumefaciens Ti-plasmid.Nature,303, 179–80.

    Article  Google Scholar 

  • Holsters, M., de Waele, D., Depicker, A., Messens, E., Van Montagu, M. and Schell, J. (1978) Transfection and transformation ofAgrobacterium tumefaciens.Mol. Gen. Genet.,163, 181–7.

    Article  PubMed  Google Scholar 

  • Horlow, C. (1991) Création de lignées de tabac mâle stériles cytoplasmiques par androgenèsein situ. Thèse de doctorat, Université de Paris-Sud, Centre d'Orsay, France.

    Google Scholar 

  • Inzé, D., Follin, A., van Lijsebettens, M., Simoens, C., Genetello, C., Van Montagu, M. and Schell, J. (1984) Genetic analysis of the individual T-DNA genes ofAgrobacterium tumefaciens: further evidence that two genes are involved in indole-3-acetic acid synthesis.Mol. Gen. Genet.,194, 265–74.

    Article  Google Scholar 

  • Jefferson, R. A., Kavanagh, T. A. and Bevan, M. W. (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.EMBO J.,6, 3901–7.

    PubMed  Google Scholar 

  • Jones, J. D. G., Gilbert, D. E., Grady, K. L., and Jorgensen, R. A. (1987) T-DNA structure and gene expression in petunia plants transformed byAgrobacterium tumefaciens C58 derivatives.Mol. Gen. Genet.,207, 478–85.

    Article  Google Scholar 

  • Jorgensen, R., Snyder, C. and Jones, J. D. G. (1987) T-DNA is organized predominantly in inverted repeat structures in plants transformed withAgrobacterium tumefaciens C58 derivatives.Mol. Gen. Genet.,207, 471–7.

    Article  Google Scholar 

  • Karlin-Neumann, G. A., Brusslan, J. A., and Tobin, E. M. (1991) Phytochrome control of thetms2 gene in transgenicArabidopsis: A strategy for selecting mutants in the signal transduction pathway.The Plant Cell,3, 573–82.

    Article  PubMed  Google Scholar 

  • Koncz, C. and Schell, J. (1986) The promoter of TL-DNA gene 5 controls the tissue-specific expression of chimaeric genes carried by a novel type ofAgrobacterium binary vector.Mol. Gen. Genet.,204, 383–96.

    Article  Google Scholar 

  • Maniatis, T., Fritsch, E. F., and Sambrook J. (1982)Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY. USA: Cold Spring Harbor Laboratory Press.

    Google Scholar 

  • Michel, M. F., Brasileiro, A. C. M., Depierreux, C., Otten, L., Delmotte, F. and Jouanin, L. (1990) Identification of differentAgrobacterium strains isolated from the same forest nursery.Appl. Environ. Microbiol.,56, 3537–45.

    Google Scholar 

  • Miller, J. H. (1972)Experiments in Molecular Genetics. Cold Spring Harbor, NY, USA: Cold Spring Harbor Laboratory Press.

    Google Scholar 

  • Murashige, T. and Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture.Physiologia Plantarum,15, 473–97.

    Google Scholar 

  • Offringa, I. A., Melchers, L. S., Regensburg-Tuink, A. J. G., Costantino, P., Schilperoort, R. A. and Hooykaas, P. J. J. (1986) Complementation ofAgrobacterium tumefaciens tumor-inducing aux mutants by genes from the TR-region of the Ri plasmid ofAgrobacterium rhizogenes.Proc. Natl Acad. Sci., USA,83, 6935–9.

    Google Scholar 

  • Pelletier, G., Ferault, M., Goujaud, J., Vedel, F. and Caboche, M. (1987) The use of rootless mutants for the screening of spontaneous androgenetic and gynogenetic haploids inNicotiana tabacum: evidence for the direct transfer of cytoplasm.Theor. Appl. Genet. 75, 13–5.

    Article  Google Scholar 

  • Pelletier, G., Férault, M., Lancelin, D. and Boulidard, L. (1989) c.m.s.Brassica oleracea cybrids and their potential for hybrid seed production. InScience for Plant Breeding (book of poster abstracts, part I). Proc 12th Cong Eucarpia. Göttingen Germany, Feb 27th-March 4th, pp. 11–7.

  • Schröder, G., Waffenschmidt, S., Weiler, E. W. and Schröder, J. (1984) The T-region of Ti plasmids codes for an enzyme synthesizing indole-3-acetic acid.Eur. J. Biochem.,138, 387–91.

    Article  PubMed  Google Scholar 

  • Tepfer, M. and Casse-Delbart, F. (1987)Agrobacterium rhizogenes as a vector for transforming higher plants.Microbiol. Sci.,4, 24–8.

    PubMed  Google Scholar 

  • Thomashow, M. F., Hungly, S., Buchholz, W. G. and Thomashow, L. S. (1986) Molecular basis for the auxin-independent phenotype of crown gall tumour tissues.Science,231, 616–8.

    PubMed  Google Scholar 

  • Thompson, K. F. (1956) Production of haploid plants of marrowstem kale.Nature,178, 748.

    PubMed  Google Scholar 

  • While, F. F., Taylor, B. H., Huffman, G. A., Gordon, M. P. and Nester, E. W. (1985) Molecular and genetic analysis of the transferred DNA regions of the root-inducing plasmid ofAgrobacterium rhizogenes.J. Bacteriol.,164, 33–44.

    PubMed  Google Scholar 

  • Williams, P. (1985)Crucifer Genetics Cooperative Resource Book., Univ. of Wisconsin, USA.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Béclin, C., Charlot, F., Botton, E. et al. Potential use of theaux2 gene fromAgrobacterium rhizogenes as a conditional negative marker in transgenic cabbage. Transgenic Research 2, 48–55 (1993). https://doi.org/10.1007/BF01977680

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01977680

Keywords

Navigation