Selectable marker-free transgenic orange plants recovered under non-selective conditions and through PCR analysis of all regenerants View Full Text


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Article Info

DATE

2010-09

AUTHORS

A. Ballester, M. Cervera, L. Peña

ABSTRACT

Selectable marker (SM) genes have been considered necessary to achieve acceptable rates in the generation of transgenic plants. Genes encoding antibiotic or herbicide resistance are widely used for this purpose. In most cases, once transgenic plants have been regenerated, permanence of SM genes in the plant genome is no longer necessary, and it becomes a matter of public concern. Moreover, the removal of SM genes from transgenic plants could facilitate gene stacking through successive transformations, particularly when the availability of these markers is rather limited for most crop plants. In the genus Citrus, with highly heterozygotic species of long generation cycles, methods implying the segregation and removal of marker transgenes in the progeny are not feasible. Here, we have evaluated the direct production of SM-free citrus plants under non-selective conditions, using a “clean” binary vector carrying only the transgene of interest, and through the recovery of transformants by polymerase chain reaction (PCR) analysis of all regenerated shoots. The response of two different citrus genotypes, Carrizo citrange (intergeneric hybrid of C. sinensis L. Osb. X Poncirus trifoliata L. Raf.) and Pineapple sweet orange (C. sinensis L. Osb.), was evaluated. Our results indicate that, in this system, the competence between transgenic and non-transgenic cells is the main factor determining final transgenic regeneration frequencies. For Carrizo citrange, no transgenic plant could be recovered. For Pineapple sweet orange, marker-free transformation efficiency was 1.7%, paving the way for the viable production of orange transformants carrying only the transgene(s) of interest. More... »

PAGES

329-336

References to SciGraph publications

  • 2007-11. Direct creation of marker-free tobacco plants from agroinfiltrated leaf discs in PLANT CELL REPORTS
  • 1993-07. NewAgrobacterium helper plasmids for gene transfer to plants in TRANSGENIC RESEARCH
  • 2003-08. Marker-free transgenic plants in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 1999-07. Green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants in THEORETICAL AND APPLIED GENETICS
  • 1998-04. Analysis of mannose selection used for transformation of sugar beet in MOLECULAR BREEDING
  • 2003-04. A transformation method for obtaining marker-free plants of a cross-pollinating and vegetatively propagated crop in NATURE BIOTECHNOLOGY
  • 2008-06. Evaluation of selection strategies alternative to nptII in genetic transformation of citrus in PLANT CELL REPORTS
  • 2000-12. Agrobacterium-mediated sorghum transformation in PLANT MOLECULAR BIOLOGY
  • 2009-03. Production of selectable marker-free transgenic tobacco plants using a non-selection approach: chimerism or escape, transgene inheritance, and efficiency in PLANT CELL REPORTS
  • 2007-01. Efficient production of transgenic citrus plants using isopentenyl transferase positive selection and removal of the marker gene by site-specific recombination in PLANT CELL REPORTS
  • 2000-11. An alternative method for the genetic transformation of sweet organce in IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY - PLANT
  • 2002-06. Regeneration of transgenic citrus plants under non selective conditions results in high-frequency recovery of plants with silenced transgenes in MOLECULAR GENETICS AND GENOMICS
  • 1998-12. Agrobacterium-mediated transformation of citrange: factors affecting transformation and regeneration in PLANT CELL REPORTS
  • 2003-05. Stable wheat transformation obtained without selectable markers in PLANT MOLECULAR BIOLOGY
  • 2004. Agrobacterum-Mediated Transformation of Citrus in TRANSGENIC CROPS OF THE WORLD
  • 1983-09. A plant DNA minipreparation: Version II in PLANT MOLECULAR BIOLOGY REPORTER
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    URI

    http://scigraph.springernature.com/pub.10.1007/s11240-010-9737-1

    DOI

    http://dx.doi.org/10.1007/s11240-010-9737-1

    DIMENSIONS

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