The Initiation of Auxin Autonomy in Tissue from Tobacco Plants Carrying the Auxin Biosynthesizing Genes from the T-DNA of Agrobacterium tumefaciens

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Abstract
Tobacco (Nicotiana tabacum cv Havana 425) plants containing the indole-3-acetic acid biosynthesizing genes (1 and 2) from the T-DNA of Agrobacterium tumefaciens strain T37-ADH2 (mutated at the cytokinin biosynthesis gene 4) were used to study the physiological basis of the suppression and reinitiation of the auxin autonomous phenotype. The plants, though normal in appearance and cross-fertile with nontransformed, wild type tobacco, are shown to contain multiple copies of genes 1 and 2. Plants carrying these genes respond to inoculation by Agrobacterium strains mutated at genes 1 and 2 in a virulent fashion. Despite the presence and potential in planta activity of these genes, pith explants from such plants require auxin or tryptophan for growth in vitro, as does wild type tobacco. In both cases the indole-3-acetic acid levels increase rapidly in pith explants cultured on tryptophan-containing medium. However, only the tissues containing genes 1 and 2 grow subsequently on auxin-free medium and accumulate indole-3-acetic acid to levels that support growth. The capacity of such tissues to utilize naphthalene acetamide as an auxin suggests that gene 2 is rapidly activated during the reinitiation process.