Advances in selectable marker genes for plant transformation

Sundar, Isaac K.; Sakthivel, N.

doi:10.1016/j.jplph.2008.08.002

Cited by 74 publications

(50 citation statements)

References 144 publications

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“…DNA vectors used in plant transformation usually contain selectable marker genes, such as those that confer antibiotic or herbicide resistance, to facilitate the selection of transformed cells (Sundar and Sakthivel, 2008). However, the retention of such marker genes in transgenic plants is undesirable because of their possible toxicity or allergenicity to humans and their unpredictable hazards to the environment (Miki and McHugh, 2004;Ramessar et al, 2007;Yau and Stewart, 2013).…”

mentioning

confidence: 99%

Efficient Generation of Marker-Free Transgenic Rice Plants Using an Improved Transposon-Mediated Transgene Reintegration Strategy

Gào

Zhou

et al. 2014

Plant Physiol.

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(S.Q.).Marker-free transgenic plants can be developed through transposon-mediated transgene reintegration, which allows intact transgene insertion with defined boundaries and requires only a few primary transformants. In this study, we improved the selection strategy and validated that the maize (Zea mays) Activator/Dissociation (Ds) transposable element can be routinely used to generate marker-free transgenic plants. A Ds-based gene of interest was linked to green fluorescent protein in transfer DNA (T-DNA), and a green fluorescent protein-aided counterselection against T-DNA was used together with polymerase chain reaction (PCR)-based positive selection for the gene of interest to screen marker-free progeny. To test the efficacy of this strategy, we cloned the Bacillus thuringiensis (Bt) d-endotoxin gene into the Ds elements and transformed transposon vectors into rice (Oryza sativa) cultivars via Agrobacterium tumefaciens. PCR assays of the transposon empty donor site exhibited transposition in somatic cells in 60.5% to 100% of the rice transformants. Marker-free (T-DNA-free) transgenic rice plants derived from unlinked germinal transposition were obtained from the T1 generation of 26.1% of the primary transformants. Individual marker-free transgenic rice lines were subjected to thermal asymmetric interlaced-PCR to determine Ds(Bt) reintegration positions, reverse transcription-PCR and enzyme-linked immunosorbent assay to detect Bt expression levels, and bioassays to confirm resistance against the striped stem borer Chilo suppressalis. Overall, we efficiently generated marker-free transgenic plants with optimized transgene insertion and expression. The transposon-mediated marker-free platform established in this study can be used in rice and possibly in other important crops.

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confidence: 99%

Efficient Generation of Marker-Free Transgenic Rice Plants Using an Improved Transposon-Mediated Transgene Reintegration Strategy

Gào

Zhou

et al. 2014

Plant Physiol.

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“…Additionally, the use of non-metabolizable genes instead of antibiotic and herbicide resistance genes as selective agents provides a suitable alternative and would satisfy public concerns about their dissemination into the environment and potential effect to consumers. This PMI positive selection system has been shown to be more effective than using kanamycin in many plant transformation protocols (Sundar & Sakthivel, 2008) but this did not seem to be the case in citrus.…”

Section: Adequate Selection Conditionsmentioning

confidence: 99%

Citrus Transformation: Challenges and Prospects

Febres¹,

Fisher²,

Khalaf³

et al. 2011

Genetic Transformation

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“…Genes as cki1 or the most employed isopentenyl transferase ipt gene encoding the enzyme IPT, catalyze the synthesis of isopentyl-adenosine-5-monophosphate, which is the first step in cytokinin biosynthesis (Miki & McHugh, 2004). This gene modify the endogenous balance between cytokinins and auxins, stimulating cell division and differentiation of the cells that promote an altered morphology, development and physiology of transgenic plants (Sundar & Sakthivel, 2008). Some authors observed that the ipt gene improved transformation efficiency in apricot leaf explants in comparison with the selection through nptII (López-Noguera et al, 2009).…”

Section: Selection Systems a Critical Stepmentioning

confidence: 99%