The activation of the Agrobacterium virulence system is known to be induced by certain phenolic compounds. We have tested the vir-inducing ability of fifty compounds, by using a virB-lacZ gene fusion, and analysed the relationship between structure and activity of these compounds. In this way we have identified several new vir-inducers: coniferylalcohol, 3,5-dimethoxy-4-hydroxybenzene, homovanillic acid, ferulic acid, 3-ethoxy-4-hydroxybenzaldehyde and guaiacol, all of which are compounds with strong or moderate activity and four compounds with weak vir-inducing activity. In view of the specificity of vir-inducers, our data extended observations of others and enabled us to define the specific structural features of a vir-inducer molecule. In addition we show here that induction of the octopine Ti vir-genes is (i) optimal at 29 degrees C and totally abolished at 37 degrees C, and (ii) strongly inhibited at low concentrations of sodium chloride. The implications for plant transformation are discussed.

Tumours are induced by Agrobacterium tumefaciens on a variety of plants. The virulence determinants of A. tumefaciens reside on a large tumour-inducing (Ti) plasmid. This plasmid carries two regions essential for tumour induction, namely the T region and the Vir region. During infection the T region is transferred to the plant cell, where it becomes stably integrated in one of the host chromosomes as T-DNA. Expression of T-DNA leads to the production of the plant hormones auxin and cytokinin, as well as to the synthesis of specific amino-acid derivatives termed opines. Agrobacterium strains are classified according to the types of opines produced by the tumours they induce. The Vir region contains genes that are expressed in the bacterium and are required for T-DNA transfer to plant cells, and several other genes that affect the efficiency of transfer and the host range. Vir regions from different Ti plasmids may vary slightly in the genes they contain: for instance, the virF gene, which is present in the Vir-region of octopine Ti plasmids, is absent from nopaline Ti plasmids. Mutation of the virF gene leads to a weakened virulence of octopine strains on tomato and Nicotiana glauca (shrub tobacco). Nopaline strains are strongly attenuated in N. glauca compared with octopine strains because of the absence of the virF virulence gene from the Ti plasmid in nopaline strains. The virF gene product may be transferred to and be active in plant cells. Here we isolate transgenic N. glauca plants in which the virF coding sequence is expressed using the cauliflower mosaic virus 35S promoter. The presence of the VirF protein converts the non-host N. glauca into a host for tumour formation by A. tumefaciens nopaline strains and octopine virF mutants. Our results indicate that certain virulence gene products such as the VirF protein may be transferred to plant cells during tumour induction, where they function as mediators of T-DNA transfer.

In this paper we provide information indicating that the agropine-type root-inducing (Ri) plasmid pRil855 ofAgrobacterium rhizogenes contains functional genes for auxin production (aux) in the right transferred DNA (T-DNA) region (TR-region). These genes were cloned and introduced into the T-region of the tumor-inducing (Ti) plasmids of mutants of Agrobacterium tumefaciens carrying an aux mutation. Depending on the Ri aux gene present, the oncogenicity of the Ti aux-1 and/or aux-2 mutations was restored, showing that the Ri aux genes are able to complement the Ti aux genes. Agrobacterium strains with an agropine-type Ri plasmid not only cause hairy root on certain plant species, but they also induce tumors on other plant species. In this paper it is shown that a mutation in either of the aux genes in the Ri plasmid leads to a total loss of tumorigenicity and a strongly diminished rhizogenicity of the host bacterium, revealing that the aux genes are important for tumor and root induction. Agrobacterium strains containing the TR-region but not the TL (left)-region of the Ri plasmid are still tumorigenic on certain plant species but are no longer capable of hairy-root induction.

The induction of vir gene expression in different types of Agrobacterium strains shows different pH sensitivity profiles. The pH sensitivity pattern demonstrated by octopine Ti strains was similar to that of a supervirulent leucinopine Ti strain, whereas this was different from that shown by nopaline Ti strains and agropine Ri strains. Data are given which indicate that these differences are due to different properties of the virA genes of these wild types. An exceptional case was formed by strains with the limited-host-range plasmid pTiAG57 which showed AS-dependent vir induction only if reduced inoculum sizes were used and the temperature was 28 degrees C or below.

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