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Nucleic Acids Research, 1986, Vol. 14, No. 20 8073-8090
© 1986

Articles

A binary vector for transferring genomic libraries to plants

C. Simoens1, Th. Alliotte1, R. Mendel2, A. Mûller2, J. Schemann2, M. Van Lijsebettens1, J. Schell1,3, M. Van Montagu1 and D. Inzé1

1Laboratorium voor Genetica, Rijksuniversiteit Gent B-9000 Gent, Belgium 2Zentralinstitut fur Genetik and Kulturpflanzenforschung Akademie der Wissenschaften der DDR, DDR-4325 Gatersleben, GDR 3Max-Planck-Institut der Zûchtungsforshung D-5000 KÔln 30, FRG

Received August 12, 1986. Accepted September 22, 1986.

The transformation of mutant plants with a complete recombinant library derived from wild-type DNA followed by assay of transformed plants for complementation of the mutant phenotype is a promising method for the isolation of plant genes. The small genome of Arabidopsis thaliana is a good candidate for attempting this so-called shotgun transformation. We present the properties of an A. thaliana genorale library cloned in a binary vector, pC22. This vector, designed to Introduce genomic libraries Into plants, contains the oriV of the Ri plasmid pRIHR1 by which it replicates perfectly stably in Agrobacteriun. Upon transfer of the library from E. coli to A. turoefaciens large differences in transfer efficiencies of individual recombinant clones were observed. There is a direct relation between transfer efficiency and stability of the recombinant clones both in E. coli and A. turoefadens. The stability 1s independent of the insert size, but seems to be related to the nature of the insert DNA. The feasibility of shotgun transformation and problems of statistical sampling are discussed.


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