A DNA Transformation–Competent Arabidopsis Genomic Library in Agrobacterium

Abstract

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 multicellular organisms like Arabidopsis.

An Arabidopsis Homolog of the Bacterial Cell Division Inhibitor SulA Is Involved in Plastid Division[W]

“…Plantlets expressing GFP were transferred to soil in the greenhouse under short-day conditions for 2 weeks, then under long-day conditions, and their phenotype was further analyzed. Plants were transformed with AtFtsZ1-1 or AtFtsZ2-1 using the AGL-1 (Lazo et al, 1991) strain of Agrobacterium and were sown on sand for BASTA selection. Plantlets were transferred to soil after 15 d.…”

Promoter and expression studies on an Arabidopsis thaliana dehydrin gene

“…pBIl01 plasmids containing the promoter/GUS fusion constructs were transferred from Escherichia coli MV1190 strain to Agrobacterium tumefaciens strain AGL1 [19] via triparental mating with an E. coli strain that contained the mobilisation plasmid pRK2013. Transformation of Arabidopsis was carried out according to Valvekens et al [20].…”

Differential Interactions of Promoter Elements in Stress Responses of the Arabidopsis Adh Gene

“…These chimeric genes were theri subcloned as EcoRIIHindIII fragments into the polylinker 01' the binary vector pBIN19 (Bevan, 1984). AI1 binary vectors containing the chimleric gene constructs were mobilized to Agrobacterium strain L13A4404 (Ooms et al, 1982) or AGLl (Lazo et al, 1991), using pRK2013 as a helper strain for triparental matings.…”