Transformation Methods for Obtaining Marker‐Free Genetically Modified Plants

Schaart, Jan G.; Krens, Frans A.; Wolters, Anne‐Marie A.; Visser, Richard G. F.

doi:10.1002/9780470958988.ch15

Cited by 16 publications

(20 citation statements)

References 39 publications

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“…GUS (CAMBIA, Australia) or pMF2 ? GUS (Schaart et al 2010). The first vector can be considered as a model vector for protocol testing with a hygromycin resistance gene, hpt, as selectable marker and the gus gene as reporter for gene transfer both under control of the CaMV35S promoter.…”

Section: Agrobacterium-mediated Transformationmentioning

confidence: 99%

Regeneration and Agrobacterium-mediated transformation of multiple lily cultivars

Wang

Kronenburg

Menzel

et al. 2012

Plant Cell Tiss Organ Cult

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To pursue genetic improvement of lily, efficiency of both regeneration and transformation from callus cultures induced from different explants were evaluated in multiple cultivars. Thirty-five callus lines induced from filaments or styles and one control callus line derived from bulb scales of in total twenty lily cultivars representing Lilium longiflorum, Oriental 9 Trumpet and Longiflorum 9 Asiatic hybrids were maintained on a medium with 8.3 lM picloram (PIC). In this study, they were tested for their regeneration potential by transferring them onto a regeneration medium supplemented with 0.4 lM PIC and 0.044 lM 6-benzyladenine. Regeneration was obtained in all cultivars examined and the percentage varied from zero to 89 % in the 36 callus lines. Regeneration frequency was significantly influenced by the genotype (cultivar). Subculturing the calli every 4 weeks by refreshing the regeneration medium contributed positively to bulblet formation, when compared to an eight week subculture frequency. It was found that the regeneration ability generally decreased with an increasing age of the callus cultures for all cultivars. The origin of the callus (style or filament) did not lead to significant differences in regeneration frequency, but there was an interaction between callus origin and genotype. Calli of eight randomly chosen cultivars were co-cultivated with Agrobacterium tumefaciens strain AGL0 carrying binary vectors with the gus gene as reporter and putative transgenic plants were produced. GUS histochemical assays demonstrated transient and stable expression of the gus gene in both calli and regenerated lily plants. Transient expression frequencies ranged from 0.3 to 20.6 % while stable transformation was much lower, only 1.4 % as the maximum.

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Section: Agrobacterium-mediated Transformationmentioning

confidence: 99%

Regeneration and Agrobacterium-mediated transformation of multiple lily cultivars

Wang

Kronenburg

Menzel

et al. 2012

Plant Cell Tiss Organ Cult

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“…This is particularly critical when elite events intended for commercialization are developed. Schaart et al (2011) observed that late induction was more efficient in recovery of marker-free GM strawberries using the Cre-lox system. However, it is highly probable that the most suitable strategy will have to be determined on a case-by-case basis.…”

Section: The Mat Vector Systemmentioning

confidence: 96%

“…Transforming without SMGs would be the ideal way to obtain marker-free GM plants. The successful recovery of GM plants without the use of SMGs has been reported for several plant species including Arabidopsis, potato, tobacco, lime, peanut, triticale, cassava, barley (reviewed by Manimaran et al, 2011;Schaart et al, 2011;Chong-Pérez and Angenon, 2013), alfalfa (Ferradini et al, 2011b), apple (Malnoy et al, 2010), orange (Ballester et al, 2010), Prunus (Petri et al, 2011), wheat (Liu et al, 2011) and tomato (Xin and Guo, 2012). In most cases putative transformed plants were screened by Polymerase Chain Reaction (PCR) for the presence of the transgene.…”

Section: F Marker-less Transformationmentioning

confidence: 99%

“…Application of germlinespecific promoters allows more efficient inheritance of the recombined loci. Schaart et al (2011) also developed a vector system where the recombinase activity is regulated at the level of its subcellular location rather than expression (the recombinase protein being able to enter the nucleus only after addition of a chemical).…”

Section: Site-specific Recombinationmentioning

confidence: 99%

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Alternatives to Antibiotic Resistance Marker Genes forIn VitroSelection of Genetically Modified Plants – Scientific Developments, Current Use, Operational Access and Biosafety Considerations

Breyer

Kopertekh

Reheul

2014

Critical Reviews in Plant Sciences

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“…with their own promoters and introns (Jacobsen & Schouten 2007). In addition, the use of additional genes for selecting transformants should be avoided or such selection genes should be removed by inducible site-specific recombination methods (Schaart et al 2011). Intragenesis likewise involves the use of genes (sequences) from cross-compatible species and marker-free transformation, but the gene sequences are recombined with e.g.…”

Section: Technical Descriptionmentioning

confidence: 99%