Research Paper | Published:

A DNA Transformation–Competent Arabidopsis Genomic Library in Agrobacterium

Bio/Technology volume 9, pages 963967 (1991) | Download Citation



We have constructed a nuclear genomic library from the cruciferous plant Arabidopsis thaliana ecotype Columbia in a cosmid vector, pLZO3, and a host organism, Agrobacterium tumefaciens AGL1, which can directly DNA–transform the parent organism, Arabidopsis. The broad host range cosmid pLZO3 carries a gentamicin acetyltransferase gene as bacterial selective marker and tandem, chimeric neomycin and streptomycin phosphotransferase genes as plant selective markers. Agrobacterium AGL1 carries the hypervirulent, attenuated tumor–inducing plasmid pTiBo542 from which T–region DNA sequences have been precisely deleted, allowing optimal DNA transformation of many dicotyledonous plants. Agrobacterium AGL1 also carries an insertion mutation in its recA general recombination gene, which stabilizes the recombinant plasmids. The Arabidopsis genomic library consists of some 21,600 clones gridded onto 96–well microtiter dishes and, if random, carries at least three genomic equivalents. When probed for the presence of several Arabidopsis low copy–number genes, the genomic library seems representative. As with the unicellular organisms Escherichia coli and Saccharomyces cerevisiae, this DNA transformation competent genomic library should expedite gene isolation, by gene rescue, in multicel–lular organisms like Arabidopsis.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1.

    , and 1988. Agrobacterium tumefaciens mediated transformation of Arabidopsis root explants using kanamycin selection. Proc. Natl. Acad. Sci. USA 85: 5536–5540.

  2. 2.

    , , , , , , , and 1991. T-DNA integration: a mode of illegitimate recombination in plants. EMBO J. 10: 697–704.

  3. 3.

    and , 1981. Somaclonal variation: a novel source of variability from tell cultures for plant improvement. Theor. Appl. Genet. 60: 197–214.

  4. 4.

    and 1983. Single gene mutations in tomato plants regenerated from tissue culture. Science 221: 949–951.

  5. 5.

    and 1988. The chromosomal basis of somaclonal variation. Ann. Rev. Plant Physiol. 39: 413–437.

  6. 6.

    , and , 1986. Overdrive, a T-DNA transmission enhancer on the Agrobacterium tumefaciens tumor-inducing plasmid. EMBO J. 5: 1137–1141.

  7. 7.

    1987. Gene tandem mediated selection of coliphage λ-receptive Agrobacterinm, Pseudomonas, and Rhizobium strains. Proc. Natl. Acad. Sci. U.S.A. 84: 3334–3338.

  8. 8.

    , and Extension of bacteriophage λ host range: selection cloning and characterization of a constitutive λ receptor gene. Proc. Natl. Acad. Sci. U.S.A. 81: 6080–6084.

  9. 9.

    , , and 1980. Broad host range DNA cloning system for Gram-negative bacteria: Construction of a gene bank of Rhizobium meliloti. Proc. Natl. Acad. Sci. USA 77: 7347–7351.

  10. 10.

    , , and 1986. The hypervirulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of T DNA. J. Bacteriol. 168: 1291–1304.

  11. 11.

    , , , , and 1984. Restriction endonuclease map of pTiBo542, a potential Ti plasmid vector for genetic engineering of plants. Bio/Technology 2: 702–709.

  12. 12.

    and 1986. The genetic and transcriptional organization of the vir region of the A6 Ti plasmid of Agrobacterium tumefaciens. EMBO. J. 5: 1445–1454.

  13. 13.

    , , and 1985. Identification of the signal molecules produced by wounded plant cells that activate T-DNA transfer in Agrobacterium tumefaciens. Nature 318: 624–629.

  14. 14.

    , and 1986. Generation of single-stranded T-DNA molecules during the initial stages of T-DNA transfer from Agrobacterium tumefaciens to plant cells. Nature 322: 706–712.

  15. 15.

    and 1988. Specialized binary vector for plant transformation: expression of the Arabidopsis thaliana AHAS gene in Nicotians tabacum. Nucleic Acids Res. 16: 10765–10782.

  16. 16.

    , and 1988. Broad host range plasmid cloning vectors for Gram-negative bacteria. Bio/Technology 10: 287–332.

  17. 17.

    , , and 1988. Improved expression of streptomycin resistance in plants due to a deletion in the streptomycin phosphotransferase coding sequence. Mol. Gen. Genet. 214: 456–459.

  18. 18.

    , and 1990. Streptomycin and lincomycin resistances are selective plastid markers in cultured Nicotiana cells. Mol. Gen. Genet 221: 245–250.

  19. 19.

    and 1986. Sulfonylurea resistant mutants of Arabidopsis thaliana. Mol. Gen. Genet. 204: 430–438.

  20. 20.

    , and 1972. Simple, rapid procedure for isolation of tobacco leaf nuclei. Anal. Biochem. 49: 48–57.

  21. 21.

    1987. Preparation of genomic DNA from plant tissue, p. 2.3.1–2.3.3. In: Current Protocols in Molecular Biology. F. Ausuhel et al. (Eds.). John Wiley and Sons, New York.

  22. 22.

    , and , Molecular Cloning, 2nd Ed Cold Spring Harbor Laboratory Press. Cold Spring Harbor, NY.

  23. 23.

    , , and 1986. In vivo repackaging of recombinant cosmid molecules for analyses of Salmonella typhimurium, Streptococcus mutans, and Mycobacterial genomic libraries. Infection and Immunity 52: 101–109.

  24. 24.

    , and 1986. Construction of a Tn5 derivative determining resistance to gentamicin and spectinomvan using a fragment cloned from R1033. Gene 48: 203–209.

Download references

Author information


  1. Department of Biology, Sinsheimer Laboratories, University of California, Santa Cruz, CA 95064.

    • Gerard R. Lazo
    • , Pascal A. Stein
    •  & Robert A. Ludwig


  1. Search for Gerard R. Lazo in:

  2. Search for Pascal A. Stein in:

  3. Search for Robert A. Ludwig in:

About this article

Publication history





Further reading