ArticlePlant molecular biology2013
The elimination of a selectable marker gene in the doubled haploid progeny of co-transformed barley plants.
Article in Plant molecular biology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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15 citing papers in PubMed, 44 citations in OpenAlex.
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- Construction of Marker-Free Genetically Modified Maize Using a Heat-Inducible Auto-Excision Vector.Genes · 2019Article
- Heritable Genomic Fragment Deletions and Small Indels in the Putative ENGase Gene Induced by CRISPR/Cas9 in Barley.Frontiers in plant science · 2017Article
- Cell layer-specific distribution of transiently expressed barley ESCRT-III component HvVPS60 in developing barley endosperm.Protoplasma · 2016Article
- Generation of Marker- and/or Backbone-Free Transgenic Wheat Plants viaFrontiers in plant science · 2016Article
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- Efficient generation of marker-free transgenic rice plants using an improved transposon-mediated transgene reintegration strategy.Plant physiology · 2015Article
- Validation of doubled haploid plants by enzymatic mismatch cleavage.Plant methods · 2013Article
- A new double right border binary vector for producing marker-free transgenic plants.BMC research notes · 2013Article
- Analysis of T-DNA integration and generative segregation in transgenic winter triticale (x Triticosecale Wittmack).BMC plant biology · 2012Article
Corrections and comments
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Authors and funding
7 authors at 3 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Following the production of transgenic plants, the selectable marker gene(s) used in the process are redundant, and their retention may be undesirable. They can be removed by exploiting segregation among the progeny of co-transformants carrying both the selectable marker gene and the effector transgene. Here we show that the doubled haploid technology widely used in conventional barley breeding programmes represents a useful means of fixing a transgene, while simultaneously removing the unwanted selectable marker gene. Primary barley co-transformants involving hpt::gfp (the selectable marker) and gus (a model transgene of interest) were produced via Agrobacterium-mediated gene transfer to immature embryos using two respective T-DNAs. These plants were then subjected to embryogenic pollen culture to separate independently integrated transgenes in doubled haploid progeny. A comparison between 14 combinations, involving two Agrobacterium strains carrying various plasmids, revealed that the highest rate of independent co-transformation was achieved when a single Agrobacterium clone carried two binary vectors. Using this principle along with Agrobacterium strain LBA4404, selectable marker-free, gus homozygous lines were eventually obtained from 1.5 per 100 immature embryos inoculated. Compared to the segregation of uncoupled T-DNAs in conventionally produced progeny, the incorporation of haploid technology improves the time and resource efficiency of producing true-breeding, selectable marker-free transgenic barley.
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