Frequent spontaneous deletions of Ri T-DNA in Agrobacterium rhizogenes transformed potato roots and regenerated plants

Caté, Clément; Loonen, Annelies E. H. M.; Ottaviani, Marie-Paule; Ennik, L.; Eldik, G. Van; Stiekema, Willem J.

doi:10.1007/bf00016506

Cited by 36 publications

(16 citation statements)

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“…Transformed roots arise as a result of the integration of T L -DNA and T R -DNA into the plant genome by the agropine strains, and its expression induces root differentiation and subsequent growth (Christey 2001). Many variations in the morphology of transformed hairy roots have been reported, and correlative studies on the integration of the genes of the T L -DNA and T R -DNA regions with the morphology of hairy roots and regenerated plants have been carried out in a few plant species (Hanisch ten Cate et al 1990;Limami et al 1998;Christey 2001).…”

Section: Introductionmentioning

confidence: 99%

Growth and terpenoid indole alkaloid production in Catharanthus roseus hairy root clones in relation to left- and right-termini-linked Ri�T-DNA gene integration

et al. 2004

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Hairy root cultures of Catharanthus roseus var. Prabal were established by infecting the leaves with Agrobacterium rhizogenes agropine-type A4 strain. Two hundred and fifty independent root clones were evaluated for growth, morphology, number of integration of Ri T-DNA genes and alkaloid contents. On the basis of growth pattern, type of branching and number of lateral roots we were able to separate the hairy root clones into four categories. However based on the integration of the Ri T(L)-DNA and T(R)-DNA genes, there were only three different categories of independent hairy root clones-C1 (rolA&B(+)/ags(+)), C2 (rolA&B(-)/ags(+)) and C3 (rolA&B(+)/ags(-)). Southern hybridization analysis revealed both single and multiple copies of T-DNA integration in the root clones. The accumulation of considerable amounts of the root-specific alkaloids ajmalicine and serpentine was observed in the presence of both the T(L)-DNA and T(R)-DNA genes (C1) and the T(L)-DNA gene (C3) alone. Two rolA&B(-) but ags(+) clones (C2) accumulated much less or only very negligible amounts of ajmalicine. The possible role of the T(L)-DNA and T(R)-DNA genes on growth and alkaloid accumulation in these root clones is discussed.

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Section: Introductionmentioning

confidence: 99%

Growth and terpenoid indole alkaloid production in Catharanthus roseus hairy root clones in relation to left- and right-termini-linked Ri�T-DNA gene integration

et al. 2004

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“…One explanation could be that the NPT II provided by the transgenic cells detoxificies the aminoglycosidic antibiotics, thus reducing their effective concentration in the tissue (6), which enables the differentiation of nontransformed cells. Another more plausible reason for the loss of expression could be due to the inactivation or silencing of introduced genes, which has frequently been observed among regenerated plants (1, 25) as well as deletion of integrated Ri T-DNA (10). The presence of the neomycinphosphotransferase enzyme activity detected in transformed plants is shown in Fig.…”

Section: Resultsmentioning

confidence: 98%

Inheritance of Kanamycin and Ethionine Resistance Introduced in Alfalfa (Medicago Sativa L.) by Gene Transfer and Cell Selection

Sarul¹,

Vlahova²,

Todorovska³

et al. 1995

Biotechnology & Biotechnological Equipment

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Fertile kanamycin resistant plants of A:fedicago sativa L. were obtained by genetic transformation with binary vector of Agrobacterium tumefaciens and were used as a fertilizer of Lethionine (ETH) resistant alfalfa variants. The dominant nature of both kanamycin and Lethionine resistant traits was demonstrated by the double resistant phenotype of obtained hybrid plants. Southern blot analysis showed that transgenic plants and self-pollinated progeny harbored the kanamycin gene, and expressed the neomycin phosphotransferase (npt) activity in vitro. The inheritance of both single (kanamycin) and double (kanamycin and Lethionine) resistance traits was anaZyzed and the way in which marker characters are segregated is discussed.

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“…While 50% of the plant clones regenerated (rolA + /rolB + /rolC + ) exhibited the typical hairy root syndrome, the remaining 50% (rolA − /rolB − /rolC − ) resembled the nontransformed plants, with a few altered characteristics [12,13]. The loss of the rolA, rolB, and rolC genes in the latter 50% of the clones in H. muticus could be due to the deletion of the rol genes during protoplast isolation or regeneration [11,50,51]. Hänisch ten Cate et al [11] concluded from their study in Solanum tuberosum that, during long-term hairy root culture and regeneration, deletions of TL-DNA and TR-DNA can occur.…”

Section: Discussionmentioning

confidence: 99%

“…The loss of the rolA, rolB, and rolC genes in the latter 50% of the clones in H. muticus could be due to the deletion of the rol genes during protoplast isolation or regeneration [11,50,51]. Hänisch ten Cate et al [11] concluded from their study in Solanum tuberosum that, during long-term hairy root culture and regeneration, deletions of TL-DNA and TR-DNA can occur. They reported the deletion of T-DNA in sister shoot lines regenerated from 4 out of 5 root clones.…”

Section: Discussionmentioning

confidence: 99%

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The Fate of Integrated Ri T-DNA rol Genes during Regeneration via Somatic Embryogenesis in Tylophora indica

2015

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The fate of integrated Ri T-DNA rol genes during regeneration via indirect somatic embryogenesis and stability of its effect on morphology and tylophorine content of Ri-transformed plants have been studied in Tylophora indica. Integration and expression of Ri T-DNA genes in transformed embryogenic callus lines derived from transformed root lines, 300 Ri-transformed somatic embryos, and 23 Ri-transformed plant lines were analysed. Fifty root lines studied showed integration and expression of four rol genes of TL-DNA. Spontaneous regeneration via indirect somatic embryogenesis was obtained from root lines that were TL + /TR − . Stable integration and expression of rol genes were observed in root lines, embryogenic callus lines, and the spontaneously induced somatic embryos. Nineteen out of the 23 Ri-transformed plant lines and their clones showed phenotypic and genetic stability over the period of 3 years. Four Ri-transformed plants were morphologically similar to nontransformed plants but showed variation with the integration and expression of the rolA gene and absence of other rol genes. Variant Ri-transformed plant line A 4 28#1-V showed highest tylophorine content (2.93 ± 0.03 mg gDW −1 ) among plant lines studied. The effects of T-DNA genes on growth, morphology, and tylophorine content of the Ri-transformed plants were stable in the long term culture.

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Frequent spontaneous deletions of Ri T-DNA in Agrobacterium rhizogenes transformed potato roots and regenerated plants

Cited by 36 publications

References 20 publications

Growth and terpenoid indole alkaloid production in Catharanthus roseus hairy root clones in relation to left- and right-termini-linked Ri�T-DNA gene integration

Growth and terpenoid indole alkaloid production in Catharanthus roseus hairy root clones in relation to left- and right-termini-linked Ri�T-DNA gene integration

Inheritance of Kanamycin and Ethionine Resistance Introduced in Alfalfa (Medicago Sativa L.) by Gene Transfer and Cell Selection

The Fate of Integrated Ri T-DNA rol Genes during Regeneration via Somatic Embryogenesis in Tylophora indica

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