AI Article Synopsis

  • Researchers found that after creating genetically modified (transgenic) barley plants, it’s better to remove selectable marker genes, as they serve no purpose once the desired traits are established.
  • They used a technique called doubled haploid technology, which helps stabilize transgenes while getting rid of unwanted markers by breeding the plants effectively.
  • The study showed that using a specific Agrobacterium strain in this process allowed them to create new lines of barley that were both true-breeding and free of the selectable marker, making the method quicker and more efficient compared to traditional genetic engineering methods.

Article 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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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3527739PMC
http://dx.doi.org/10.1007/s11103-012-9988-9DOI Listing

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