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  • Root induction  (2)
  • GUS staining  (1)
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  • 1
    ISSN: 1573-5028
    Keywords: Agrobacterium rhizogenes ; gene expression ; GUS staining ; in situ hybridization ; rolA gene ; tobacco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The spatial and temporal activity of the entire and individual promoter domains of the rolA gene of Agrobacterium rhizogenes was investigated and correlated with the distinctive features of the phenotypes of transgenic tobacco plants. The GUS assay was performed in the presence of an oxidative catalyst during the development of transgenic plants expressing chimeric genes containing the β-glucuronidase coding sequence under the control of the different promoter domains. In situ hybridization was also used on transgenic plants harbouring rolA under the control of the entire or deleted promoter. This paper demonstrates for the first time that the entire rolA promoter, composed of domains, A, B and C, is silent in seeds, then activated at the onset of germination in the cotyledons and in the elengation zone of the radicle and is finally expressed throughout the vegetative and floral phases. Domains B+C, which were sufficient to induce wrinkled leaves and short internodes, were active in all the stem tissues, but only in the companion cells of the phloem strands of the leaves. Domain C, which specified a dwarf phenotype with normal leaves, was weakly expressed in the stem vascular bundles and in the leaf internal phloem. These results indicate that the vascular bundles are the primary targets for the generation of the short internode phenotype. Furthermore, the local expression of rolA in the stem vascular bundles induced a size reduction of the surrounding parenchyma cells, suggesting the existence of some diffusible factor(s) associated with the expression of the rolA gene.
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  • 2
    ISSN: 1617-4623
    Keywords: Agrobacterium rhizogenes ; Root induction ; T-DNA ; Transformation ; Tobacco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Disarmed plant transformation vectors were used to assay the ability of subfragments of the T-regions of the Ri plasmid of agropine-type strain A4 of Agrobacterium rhizogenes to induce proliferation of transformed roots on tobacco leaf fragments. We have shown that a 6 kb region of TR-DNA, bearing the presumptive auxin synthesis genes, is capable of inducing transformed roots with an essentially normal phenotype as had been shown previously with the entire TR-region. A 1.9 kb fragment of the 20 kb TL-region is suffcient to induced transformed roots in the absence of exogenous hormones. These roots grow profusely on hormone-free medium, as is typical of roots transformed by the intact TL-DNA.
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  • 3
    ISSN: 1617-4623
    Keywords: Agrobacterium rhizogenes ; T-DNA ; Root induction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary To analyse the respective role of TL- and TR-DNA in root induction by agropine-type Agrobacterium rhizogenes Ri plasmids, deletions covering the TL- or the TR-regions were constructed in vitro and introduced into pRiA4 by marker exchange. Each T-region of pRiHRI was also cloned separately on an independent replicon and used in a binary system with the virulence functions of either an Ri or a Ti plasmid provided in trans. Transformed roots were induced on tobacco and tomato explants by TL-DNA as well as by TR-DNA, suggesting that agropine type Ri plasmids from strains A4 and HRI can induce root proliferation by two independent transformation mechanisms. The root induction by the TR-DNA is probably due to auxin biosynthesis by gene products of aux loci homologous to the tms genes of Ti plasmid T-DNA. The molecular mechanism of root proliferation induced by the TL-DNA is probably equivalent to that of mannopine type Ri plasmid T-DNA.
    Type of Medium: Electronic Resource
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